Salidroside derivative with HIF-1alpha protein inhibitory activity as well as synthesis and application of salidroside derivative

Abstract

The invention provides a salidroside derivative with HIF-1alpha inhibitory activity as well as a synthesis method and application of the salidroside derivative. According to the present invention, the dual luciferase reporter gene experiment results show that the derivative has significant transcription inhibition activity on HIF-1 [alpha] HRE, and has low astrocyte C8-D1A cell toxicity; the compound is simple in preparation method, raw materials are cheap and easy to obtain, and the compound is expected to become a novel medicine for preventing and / or treating acute and chronic altitude sickness such as acute altitude encephaledema, acute altitude pulmonary edema, altitude polycythemia, altitude pulmonary hypertension and altitude decline related to the HIF-1alpha protein.

Description

Technical Field

[0001] The present invention belongs to the fields of medicinal chemistry and pharmaceutical technology, and particularly relates to a salidroside derivative with HIF-1α protein inhibitory activity, and its synthesis and application. Background Art

[0002] The plateau environment above 2,500 meters above sea level has characteristics such as low pressure, low oxygen, cold, dryness, and strong radiation. For every 1,000-meter increase in altitude, the atmospheric pressure drops by about 10 kPa, and the oxygen partial pressure drops by about 10% (N. Engl. J. Med. 2022, 386(4): 364 - 373.). Altitude sickness is a series of pathological changes that occur when the human body rapidly enters the plateau from the plain area or is exposed to the plateau environment for a long time. The body's compensatory function is disordered due to hypoxia, resulting in acute mild altitude sickness such as dizziness, headache, palpitation, shortness of breath, loss of appetite, fatigue, nausea, and insomnia. After short-term adaptation or symptomatic treatment, its symptoms and signs gradually disappear. However, those with poor acclimatization will further induce acute altitude sickness, including acute high altitude cerebral edema, acute high altitude pulmonary edema, and mixed acute altitude sickness, which are characterized by high incidence, high fatality rate, and great harm (N. Engl. J. Med. 2013, 368(24): 2294 - 2302.). Chronic altitude sickness refers to a clinical syndrome caused by the loss of plateau adaptation after the body has lived or inhabited the plateau for a long time, including high altitude polycythemia, high altitude pulmonary hypertension, abnormal high altitude blood pressure, high altitude heart disease, high altitude decline syndrome, and mixed chronic altitude sickness. Clinically, symptomatic treatment is mainly used, and specific drugs are lacking, which seriously endangers the lives and health of people in the plateau area and the development of economic construction. The research and development of drugs for the prevention and treatment of altitude sickness are urgent (Pharmaceutics. 2024, 16(11): 1375.).

[0003] Rhodiola rosea L. is a perennial herb of the genus Rhodiola in the family Crassulaceae, mainly growing in the plateau area at an altitude of 1,800 m to 2,500 m, and is known as the "ginseng on the plateau" and other reputations (Front. Pharmacol. 2022, 13: 974775.). Salidroside is one of the main active ingredients of Rhodiola rosea, and has biological activities such as neuroprotection, cardioprotection, anti-inflammatory, and antioxidant stress. It upregulates the expression of tight junction proteins such as ZO-1 and occluding, reduces LD, LDH, and increases Na + -K + -ATPase, Ca 2+ -Mg 2+The content of ATPase, etc., inhibits the NF-κB / NLRP3 inflammatory pathway, effectively reducing low-pressure hypoxia-induced cerebral oxidative stress injury, inflammatory response, and blood-brain barrier disruption, and has a preventive and therapeutic effect on high-altitude cerebral edema and high-altitude pulmonary edema (Eur. J. Pharmacol. 2022, 925: 175015.). However, salidroside has strong water solubility, poor cell membrane permeability, low oral bioavailability, fast metabolism, and a large waste of Rhodiola resources in plant extraction (Biomed. Pharmacother. 2020, 129: 110458.).

[0004] Hypoxia-inducible factor-1α (HIF-1α) is a key oxygen homeostasis regulatory protein in cells. Under normoxic conditions, the HIF-1α protein is hydroxylated by prolyl hydroxylase (PHDs), and then degraded through the ubiquitin-proteasome pathway mediated by the tumor suppressor protein pVHL (J. Med. Chem. 2019, 62(12): 5725–5749.). When the human body is acutely or chronically exposed to the high-altitude hypoxic environment, the HIF-1α protein is overexpressed in tissues such as the brain and lungs. It binds to the HIF-1β protein in the cell nucleus to form a dimer, activating the transcription of the HIF-1 gene, inducing the overexpression of downstream vascular endothelial growth factor (VEGF) and the degradation of matrix metalloproteinase (MMP-9), causing changes in vascular permeability and blood-brain barrier disruption; inducing the inactivation of Na + / K + -ATPase, resulting in the swelling of glial cells and endothelial cells; regulating the overexpression of inflammation- and oxidative stress-related factors such as nitric oxide synthase, inflammatory factors (NF-κB, IL-6, and TNF-α, etc.), and reactive oxygen species (ROS), etc., inducing high-altitude cerebral edema and high-altitude pulmonary edema (High Alt. Med. Biol. 2024, 25(4): 326-336.). The HIF-1α protein also promotes the overexpression of downstream erythropoietin (EPO) transcription. Long-term exposure to the high-altitude environment leads to excessive proliferation of red blood cells, thickening of the blood, and induction of chronic altitude diseases such as high-altitude polycythemia (Front. Med. 2024, 11: 1448654.). Therefore, through structural modification, it is of great research significance and has a far-reaching market application prospect to discover salidroside derivatives with better pharmacological activity and more stable metabolism that target and inhibit the HIF-1α protein for the development of preventive and / or therapeutic drugs for acute and chronic altitude diseases. Summary of the Invention

[0005] The purpose of the present invention is to provide a salidroside derivative having an inhibitory activity on hypoxia-inducible factor-1α (HIF-1α) protein, and its use as a drug for altitude diseases related to HIF-1α overexpression, especially in the preventive and / or therapeutic drugs for acute and chronic altitude diseases such as acute high-altitude cerebral edema, acute high-altitude pulmonary edema, high-altitude polycythemia, high-altitude pulmonary hypertension, and altitude decline syndrome, etc.

[0006] The first aspect of the present invention provides a salidroside derivative having hypoxia-inducible factor-1α (HIF-1α) protein inhibitory activity, or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, polymorph, metabolite or isotope-labeled compound thereof, and the structure of the derivative is shown in formula (I):

[0007]

[0008] In formula (I):

[0009] Ring A is absent, or a substituted / unsubstituted benzene ring or pyridine ring; preferably one or more of absent, benzene ring or pyridine ring;

[0010] X is selected from oxyacetyl ester group, acryloyl ester group, oxyacetyl amino group, 2-butenamide group, acetamide group, propionamide group, acrylamide group, 1-acrylamido group, carbamoyl group, acetamido group, hydrazide group, propenyl group, mercaptoacetamide group, oxyacetamide group, oxyacrylamide group, 3-(1-propenyl)-3,4,5-triazolyl group; preferably at least one group selected from acryloyl ester group, acrylamide group, acetamide group, oxyacetamide group;

[0011] R 1 is hydrogen or substituted with a glucosyl group; preferably substituted with a glucosyl group;

[0012] R 2 is hydrogen or substituted with 2-fluoro, 4-methoxy-3-hydroxy, 4-hydroxy, 3-methoxy-4-hydroxy; preferably substituted with 2-fluoro;

[0013] n is 0 or 1, indicating absent or containing a saturated ethyl group; n is preferably 1.

[0014] According to some embodiments of the present invention, is selected from the following fragments:

[0015]

[0016] Preferably, is selected from

[0017] is selected from the following fragments:

[0018]

[0019] Preferably, is selected from

[0020] R 1 is selected from the following fragments:

[0021] H,

[0022] Preferably, R 1 is selected from

[0023] selected from the following fragments:

[0024]

[0025] Preferably, is selected from

[0026] According to some embodiments of the present invention, the salidroside derivative of the above formula (I) may specifically be a compound of the following formula (II), (III), (IV) or (V):

[0027]

[0028] wherein, X and R 2 have the same substituents as above. In formula (IV) or (V), n is 0 or 1, indicating the absence or presence of one saturated ethyl group, and Y is a C atom or an N atom.

[0029] Particularly, the salidroside derivative of formula (I) of the present invention is specifically the following compound:

[0030]

[0031]

[0032] In the second aspect of the present invention, there is provided a method for synthesizing a salidroside derivative having the general formula (Ⅰ) as described in the first aspect of the present invention, including the following synthetic route:

[0033] Synthetic route one:

[0034]

[0035] R 2 The substituted phenol (II-1) reacts with methyl bromoacetate through a nucleophilic substitution reaction to obtain the intermediate II-2, and through an ester hydrolysis reaction under strong alkaline conditions to obtain R 2 substituted carboxylic acid intermediate II-3; the reaction solvent used for the nucleophilic substitution reaction is acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide, dichloromethane or ethyl acetate, preferably acetonitrile; the strong base used is selected from sodium hydroxide, lithium hydroxide, potassium hydroxide, potassium tert-butoxide, preferably sodium hydroxide;

[0036] Intermediate II-3 reacts with β-D-glucose under the action of a coupling reagent to prepare the compound of formula (II) through the Mitsunobu reaction; the coupling reagent used is selected from diethyl azodicarboxylate, diisopropyl azodicarboxylate, dimethyl azodicarboxylate, di-tert-butyl azodicarboxylate, bis(4-chlorobenzyl) azodicarboxylate, and preferably diethyl azodicarboxylate;

[0037] The definitions of the groups in the above synthetic route are as described in the first aspect of the present invention.

[0038] Synthetic route two:

[0039]

[0040] p-Nitrophenol (III-1) undergoes a nucleophilic substitution reaction with (3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate, and the coupling reagent used in the nucleophilic substitution reaction is boron trifluoride diethyl ether complex, boron trifluoride acetonitrile complex, or a complex of lithium aluminum hydride and boron trifluoride diethyl ether, preferably boron trifluoride diethyl ether complex; Intermediate III-2 is reduced under the action of a reducing agent to obtain the nitro reduction product Intermediate III-3, and the reducing reagent used is iron powder, zinc powder, sodium borohydride, lithium aluminum hydride, or hydrogen, preferably iron powder or hydrogen; Intermediate III-3 further reacts with R 2 substituted phenyl acid through an amide condensation reaction to obtain Intermediate III-4, and the coupling reagent used in the amide condensation reaction is selected from 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), preferably HATU;

[0041] p-Acetoxybenzoic acid (III-5) reacts with R 2 substituted phenylamino side chain through an amide condensation reaction to obtain III-6, and the coupling reagent used in the amide condensation reaction is selected from HATU, EDCI, O-benzotriazol-N,N,N',N'-tetramethyluronium tetrafluoroborate (TBTU), preferably HATU; Intermediate III-6 is deacetylated under strong basic conditions to obtain III-7, and the strong base used is selected from sodium methoxide, lithium hydroxide, sodium hydroxide, potassium hydroxide, potassium tert-butoxide, preferably sodium methoxide; Intermediate III-7 further reacts with (3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate through a nucleophilic substitution reaction to obtain Intermediate III-8, and the coupling reagent used in the nucleophilic substitution reaction is boron trifluoride diethyl ether complex, boron trifluoride acetonitrile complex, or a complex of lithium aluminum hydride and boron trifluoride diethyl ether, preferably boron trifluoride diethyl ether complex;

[0042] R 2The substituted aniline (III-9) undergoes a nucleophilic substitution reaction with bromoacetyl bromide to obtain the intermediate III-10. The reaction solvent used can be selected from dichloromethane, acetonitrile, and tetrahydrofuran, with dichloromethane being the most preferred; the intermediate III-10 and p-methoxythiophenol or 4-methoxyphenol undergo a nucleophilic substitution reaction under strong basic conditions to obtain the intermediate III-11. The strong base can be selected from cesium carbonate, sodium carbonate, sodium hydroxide, and potassium tert-butoxide, with cesium carbonate being the most preferred; (E)-3-(4-methoxyphenoxy)acrylic acid (intermediate III-12) and different R 2 The substituted aniline undergoes an amide condensation reaction to obtain the intermediate III-13. The coupling reagent can be selected from HATU, EDCI, and TBTU, with HATU being the most preferred; different R 2 The substituted (E)-(3-azidoprop-1-en-1-yl)benzene (intermediate III-14) and 4-methoxyphenylacetylene undergo a Click reaction to obtain the intermediate III-15. The catalyst used can be selected from copper sulfate, cuprous bromide, and cuprous iodide, with copper sulfate being the most preferred; the intermediates III-11, III-13, and III-15 are prepared by a demethylation reaction to obtain the intermediate III-16. The demethylation reagent is selected from boron tribromide, hydrobromic acid, and aluminum chloride, with boron tribromide being preferred; the intermediate III-16 further undergoes a nucleophilic substitution reaction with (3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate to obtain the intermediate III-17;

[0043] The intermediates III-4, III-8, and III-17 remove the acetyl group under strong basic conditions to obtain the compound of formula (III). The strong base is selected from sodium methoxide, sodium hydroxide, potassium tert-butoxide, and cesium carbonate, with sodium methoxide being preferred;

[0044] The definitions of the groups in the above synthetic route are as described in the first aspect of the present invention.

[0045] Synthetic Route Three:

[0046]

[0047] 4-Bromophenol or 4-bromophenylethanol (IV-1) undergoes a Miyaura borylation reaction with bis(pinacolato)diboron under the action of a catalyst to obtain the borate intermediate IV-2. The catalyst is selected from [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, or tetrakis(triphenylphosphine)palladium, with [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium being preferred;

[0048] The arylamine or aryl phenolic acid (IV-3) substituted with a Y atom and R 2The substituted aryl side chain is subjected to an amide condensation reaction to obtain intermediate IV-5, or the aryl phenolic acid (IV-4) substituted with a Y atom and R 2 The substituted aryl side chain is subjected to a nucleophilic substitution reaction under strong basic conditions to obtain intermediate IV-5; the coupling reagent for the amide condensation reaction is selected from HATU, EDCI, TBTU, preferably HATU, and the strong base is selected from cesium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, preferably cesium carbonate;

[0049] Intermediate IV-5 and intermediate IV-2 are subjected to a Suzuki-Miyaura cross-coupling reaction to obtain the compound of formula (IV). The catalyst for the Suzuki-Miyaura cross-coupling reaction is selected from [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex, tetrakis(triphenylphosphine)palladium, and most preferably [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex. The strong base used in the Suzuki-Miyaura cross-coupling reaction is selected from potassium phosphate, sodium carbonate, sodium bicarbonate, cesium carbonate, and most preferably potassium phosphate;

[0050] The definitions of the various groups in the above synthetic route are as described in the first aspect of the present invention.

[0051] Synthetic route four:

[0052]

[0053] (3R,4S,5S,6R)-6-(Acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate (V-1) removes the 2-position acetyl group under basic conditions to obtain intermediate V-2. The base is selected from benzylamine, aniline, triethylamine, preferably benzylamine;

[0054] Intermediate V-2 and trichloroacetonitrile are subjected to a Pinner reaction under basic conditions to obtain intermediate V-3. The base is selected from 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), triethylamine, N,N-diisopropylethylamine, preferably DBU;

[0055] Intermediate V-3 and 4-bromophenol or 4-bromo-1-phenylethanol are subjected to a nucleophilic substitution reaction to obtain intermediate V-4. The coupling reagent used in the nucleophilic substitution reaction is boron trifluoride diethyl ether complex, boron trifluoride acetonitrile complex, preferably boron trifluoride diethyl ether complex;

[0056] Intermediate V-4 reacts with bis(pinacolato)diboron under the catalysis of a catalyst to obtain borate intermediate V-5 through Miyaura borylation reaction. The catalyst is selected from dichlorobis(1,1'-bis(diphenylphosphino)ferrocene)palladium(II), dichlorobis(1,1'-bis(diphenylphosphino)ferrocene)palladium(II) dichloromethane complex, tetrakis(triphenylphosphine)palladium(0), and preferably dichlorobis(1,1'-bis(diphenylphosphino)ferrocene)palladium(II);

[0057] Intermediate V-5 reacts with intermediate IV-5 to obtain the compound of formula (V) through Suzuki-Miyaura cross-coupling reaction. The catalyst for the Suzuki-Miyaura cross-coupling reaction is selected from dichlorobis(1,1'-bis(diphenylphosphino)ferrocene)palladium(II), dichlorobis(1,1'-bis(diphenylphosphino)ferrocene)palladium(II) dichloromethane complex, tetrakis(triphenylphosphine)palladium(0), and most preferably dichlorobis(1,1'-bis(diphenylphosphino)ferrocene)palladium(II) dichloromethane complex. The strong base used in the reaction is selected from potassium phosphate, sodium carbonate, sodium bicarbonate, cesium carbonate, and preferably potassium phosphate;

[0058] The definitions of the groups in the above synthesis route are as described in the first aspect of the present invention.

[0059] The "compound" described in the present invention includes salidroside derivatives and their pharmaceutically acceptable salts, solvates, prodrugs, stereoisomers, polymorphs, metabolites, isotope-labeled compounds, enantiomers, diastereoisomers, tautomers, or mixtures thereof.

[0060] The "pharmaceutically acceptable salt" described in the present invention can be an inorganic acid salt or an organic acid salt of the salidroside derivative represented by formula (I). The inorganic acids can include but are not limited to hydrochloric acid, sulfuric acid, hydrobromic acid, phosphoric acid, nitric acid, etc. The organic acids can include but are not limited to acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0061] The salidroside derivatives and pharmaceutically acceptable salts described in the present invention also include solvate or hydrate forms. Generally, the solvate or hydrate form is regarded as equivalent to the non-solvate form in terms of use and is included in the protection scope of the present invention. In addition, some compounds in the present invention may exist in polymorphic or amorphous crystal forms. Regardless of their physical crystal forms, they are regarded as equivalent and are included in the content of the present invention.

[0062] In a third aspect of the present invention, there is provided a pharmaceutical composition comprising the salidroside derivative of formula (I) or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, polymorph, metabolite, isotopically labeled compound, enantiomer, diastereoisomer or tautomer thereof, and at least one pharmaceutically acceptable carrier.

[0063] In a fourth aspect of the present invention, there is provided a pharmaceutical preparation comprising the salidroside derivative of formula (I) or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, polymorph, metabolite, isotopically labeled compound, enantiomer, diastereoisomer or tautomer thereof, or the pharmaceutical composition described in the third aspect of the present invention; the pharmaceutical preparation is an oral solid or liquid preparation, or a patch for topical administration.

[0064] In a fifth aspect of the present invention, there is provided the use of the salidroside derivative of formula (I), or the pharmaceutical composition, or the pharmaceutical preparation, in the preparation of a HIF-1α protein inhibitor

[0065] In a sixth aspect of the present invention, there is provided the use of the salidroside derivative of formula (I), or the pharmaceutical composition, or the pharmaceutical preparation, in the preparation of an anti-hypoxia or anti-inflammatory drug.

[0066] In a seventh aspect of the present invention, there is provided the use of the salidroside derivative of formula (I), or the pharmaceutical composition, or the pharmaceutical preparation, in the preparation of a drug for preventing or treating high altitude diseases; the high altitude diseases include:

[0067] Acute high altitude diseases: acute high altitude cerebral edema, acute high altitude pulmonary edema, mixed acute high altitude disease;

[0068] Chronic high altitude diseases: high altitude polycythemia, high altitude pulmonary hypertension, abnormal high altitude blood pressure, high altitude heart disease, high altitude deterioration syndrome, mixed chronic high altitude disease.

[0069] The salidroside derivative of the present invention has significant transcriptional inhibitory activity against HIF-1α HRE, has low in vitro cytotoxicity to astrocyte C8-D1A cells, and the preparation method of this kind of compound is simple, the raw materials are cheap and easy to obtain, and it is expected to become a new type of preventive and / or therapeutic drug for acute and chronic high altitude diseases such as acute high altitude cerebral edema, acute high altitude pulmonary edema, high altitude polycythemia, high altitude pulmonary hypertension, and high altitude deterioration syndrome related to HIF-1α protein. Experiments show that compounds 23 and 41 have a transcriptional inhibition rate of over 90% against HIF-1α HRE and low cytotoxicity, and have significant clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0070] Figure 1 It is the anti-proliferative activity of the salidroside derivative of the present invention against C8-D1A cells at a concentration of 20 μM. Detailed implementation mode

[0071] The present invention further illustrates the clarity and integrity of the research plan through specific embodiments. In particular, the scientific terms involved in the embodiments are all well-known terms in the field. The preparation methods provided in the embodiments are only exemplary descriptions and do not constitute an exclusive limitation to the present invention. Under the core concept of the present invention, simple improvements made to the preparation method are within the protection scope of the present invention. The chemical reagents and materials used in the present invention can all be obtained through commercial channels.

[0072] Example 1: Synthesis of the chemical named (2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl 2-phenoxyacetate (Compound 1) in the present invention. The synthetic route is as follows:

[0073]

[0074] Step 1: Synthesis of methyl 2-phenoxyacetate (Intermediate 1-2)

[0075] Weigh phenol (Intermediate 1-1) (6.21 g, 66 mmol), methyl bromoacetate (6.18 mL, 65 mmol), and potassium carbonate (10.84 g, 78 mmol) into a 100 mL reaction flask, add 50 mL of acetonitrile solution to dissolve, and reflux at 80 °C overnight. After the reaction is completed, evaporate the solvent, extract with ethyl acetate-water, combine the organic phases, dry over anhydrous Na2SO4, and purify by column chromatography (PE:EA = 20:1 to 5:1) to obtain 12.95 g of a pale yellow liquid with a yield > 99%. 1 H NMR (400 MHz, CDCl3) δ 7.29 (dd, J = 16.2, 7.6 Hz, 2H), 7.00 (t, J = 7.4 Hz, 1H), 6.91 (d, J = 7.8 Hz, 2H), 4.64 (s, 2H), 3.80 (s, 3H).

[0076] Step 2: Synthesis of 2-phenoxyacetic acid (Intermediate 1-3)

[0077] Weigh Intermediate 1-2 (5.00 g, 30.0 mmol) into a 100 mL reaction flask, add 50 mL of ethanol-aqueous solution (4:1) to dissolve, slowly add sodium hydroxide (4.80 g, 120.0 mmol), and reflux at 80 °C for 2 h. After the reaction is completed, evaporate the solvent, adjust the pH to 2 with 2M hydrochloric acid solution to precipitate a solid, filter by suction, wash with water, and dry to obtain 3.35 g of a white solid with a yield of 13%. 11H NMR (400 MHz, CDCl3) δ 10.13 (s, 1H), 7.34 - 7.25 (m, 2H), 7.03 (t, J = 7.2 Hz, 1H), 6.93 (d, J = 7.6 Hz, 2H), 4.69 (s, 2H).

[0078] Step 3: Synthesis of (2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl 2-phenoxyacetate (Compound 1)

[0079] Weigh intermediate 1-3 (100 mg, 2.8 mmol) into a 25 mL reaction flask, add 8 mL of tetrahydrofuran solution to dissolve it, and successively add diethyl azodicarboxylate (0.86 mL, 4.3 mmol), β-D-glucose (120 mg, 2.8 mmol), and triphenylphosphine (172 mg, 2.8 mmol). After protecting with argon, stir the reaction at room temperature for 24 h. After the reaction is completed, evaporate the solvent and purify by column chromatography (DCM:MeOH = 40:1 - 5:1) to obtain 20 mg of a pale yellow liquid with a yield of 10%. 1 1H NMR (400 MHz, CD3OD) δ 7.29 - 7.19 (m, 2H), 6.97 - 6.84 (m, 3H), 5.57 (d, J = 8.0 Hz, 1H), 4.77 - 4.63 (m, 2H), 3.85 - 3.62 (m, 2H), 3.45 - 3.39 (m, 1H), 3.38 - 3.31 (m, 2H), 3.29 - 3.24 (m, 1H); HRMS (ESI) m / z calculated for C 14 H 18 NaO8 [M + Na] + 337.0894, found 337.0890.

[0080] Example 2: Synthesis of (2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl 2-(2-fluorophenoxy)acetate (Compound 2) in the present invention

[0081]

[0082] Adapt to change the starting compound (replace the phenol starting material in Step 1 of Example 1 with a 2-fluorophenol starting material), and the remaining steps are the same as in Example 1 to obtain a white solid product with a yield of 20%. 11H NMR (400 MHz, CD3OD) δ 7.16 - 7.09 (m, 1H), 7.12 - 7.04 (m, 2H), 7.01 - 6.92 (m, 1H), 5.60 (d, J = 8.0 Hz, 1H), 4.86 (d, J = 4.4 Hz, 2H), 3.85 (dd, J = 12.0, 2.0 Hz, 1H), 3.68 (dd, J = 12.0, 4.8 Hz, 1H), 3.47 - 3.41 (m, 1H), 3.41 - 3.33 (m, 3H); HRMS (ESI) m / z calculated for C 14 H 17 FNaO8 [M+Na] + 355.0800, found 355.0767.

[0083] Example 3: Preparation of (2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl (E)-3-(2-fluorophenyl)acrylate (Compound 3) in the present invention

[0084]

[0085] Adapt to change the starting compound (replace the starting material 2-phenoxyacetic acid in step 3 of Example 1 with 2-fluorocinnamic acid), and the remaining steps are the same as in Example 1, to obtain a white solid product with a yield of 5%. 1 1H NMR (400 MHz, CD3OD) δ 8.10 (d, J = 8.8 Hz, 1H), 8.00 - 7.77 (m, 2H), 7.31 - 7.15 (m, 3H), 5.07 (d, J = 7.2 Hz, 1H), 3.92 (d, J = 12.0 Hz, 1H), 3.72 (dd, J = 12.0, 5.6 Hz, 2H), 3.58 - 3.35 (m, 1H), 3.22 (s, 1H); HRMS (ESI) m / z calculated for C 15 H 17 FNaO7 [M+Na] + 351.0851, found 351.0856.

[0086] Example 4: Preparation of 2-(2-fluorophenoxy)-N-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)acetamide (Compound 4) in the present invention, and its synthetic route is as follows:

[0087]

[0088] Step 1: Synthesis of (2R,3R,4S,5R,6S)-2-(acetoxymethyl)-6-(4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (Intermediate 4-2)

[0089] Weigh p-nitrophenol (Intermediate 4-1) (1.60 g, 11.5 mmol), (3S,4R,5R,6S)-6-(acyloxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate (3.00 g, 7.7 mmol), molecular sieve (6.00 g) into a 100 mL reaction flask, add 40 mL of dichloromethane solution to dissolve, slowly dropwise add boron trifluoride diethyl ether complex (4.85 mL, 38.4 mmol) under ice bath, and stir the reaction at room temperature for 24 h. After the reaction is completed, extract with 1 M sodium hydroxide solution, combine the organic phases, dry over anhydrous Na2SO4, and purify by column chromatography (PE:EA = 20:1 - 2:1) to obtain 2.83 g of a pale yellow liquid with a yield of 78%. 1 1H NMR (400 MHz, CDCl3) δ 8.27 - 8.18 (m, 2H), 7.21 (d, J = 9.0 Hz, 2H), 5.69 (t, J = 9.8 Hz, 1H), 5.19 (t, J = 10.0 Hz, 1H), 5.08 (dd, J = 10.4, 3.6 Hz , 1H), 4.32 - 4.17 (m, 2H), 4.09 - 4.00 (m, 2H), 2.12 - 2.00 (m, 12H).

[0090] Step 2: Synthesis of (2R,3R,4S,5R,6S)-2-(acetoxymethyl)-6-(4-aminophenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (Intermediate 4-3)

[0091] Weigh Intermediate 4-2 (2.83 g, 6.0 mmol) and ammonium chloride (260 mg, 4.8 mmol) into a 100 mL reaction flask, add 30 mL of ethanol - water solution (4:1) to dissolve, slowly add iron powder (1.69 g, 30.0 mmol), and reflux the reaction at 80 °C for 2 h. After the reaction is completed, filter while hot, evaporate the solvent, and purify by column chromatography (PE:EA = 20:1 - 1:1) to obtain 1.05 g of a pale yellow liquid with a yield of 40%. 11H NMR (400 MHz, CDCl3) δ 6.83 (d, J = 8.6 Hz, 2H), 6.62 (dd, J = 8.2, 5.8 Hz, 2H), 5.31 - 5.09 (m, 3H), 4.90 (d, J = 7.4 Hz, 1H), 4.28 (dd, J = 12.2, 5.2 Hz, 1H), 4.15 (dd, J = 12.2, 2.4 Hz, 1H), 3.78 (ddd, J = 10.0, 5.2, 2.8 Hz, 1H), 2.10 - 2.00 (m, 12H).

[0092] Step 3: Synthesis of 2-(2-fluorophenoxy)-N-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)acetamide (Compound 4)

[0093] Weigh intermediate 4-3 (328 mg, 0.7 mmol) and 2-(2-fluorophenoxy)acetic acid (152 mg, 0.9 mmol) into a 25 mL reaction flask, add 15 mL of dichloromethane solution to dissolve, successively add HATU (341 mg, 0.9 mmol) and triethylamine (0.15 mL, 1.1 mmol), and stir at room temperature for 5 h. After completion, add 20 mL of purified water, extract with ethyl acetate, combine the organic phases, dry over anhydrous Na2SO4, evaporate the solvent, add 10 mL of methanol solution to dissolve, and then slowly add sodium methoxide (63 mg, 1.2 mmol), and stir at room temperature for 1 h. After the reaction is completed, evaporate the solvent and purify by column chromatography (DCM:MeOH = 60:1 - 5:1) to obtain 148 mg of a white solid with a yield of 50%. 1 1H NMR (400 MHz, DMSO-d6) δ 10.06 (s, 1H), 7.56 - 7.48 (m, 2H), 7.29 - 7.19 (m, 1H), 7.19 - 7.06 (m, 2H), 7.03 - 6.94 (m, 3H), 5.30 (d, J = 4.8 Hz, 1H), 5.09 (d, J = 4.6 Hz, 1H), 5.02 (d, J = 5.2 Hz, 1H), 4.79 (d, J = 7.4 Hz, 1H), 4.76 (s, 1H), 4.56 (t, J = 5.8 Hz, 1H), 3.69 (ddd, J = 11.8, 5.6, 2.0 Hz, 1H), 3.51 - 3.40 (m, 1H), 3.32 - 3.11 (m, 4H); HRMS (ESI) m / z calculated for C 20 H 22 FNNaO8 [M + Na] + 446.1222, found 446.1203.

[0094] Example 5: Preparation of (E)-4-(2-fluorophenyl)-N-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)phenyl)butanamide (Compound 5)

[0095]

[0096] Adapt to change the starting compound (replace the 2-(2-fluorophenoxy)acetic acid starting material in Step 3 of Example 4 with (E)-4-(2-fluorophenyl)but-2-enoic acid starting material), and the remaining steps are the same as in Example 4 to obtain a white solid product with a yield of 50%. 1 HNMR(400MHz,DMSO-d6)δ9.94(s,1H),7.61(td,J=7.8,1.6Hz,1H),7.54-7.47(m,2H),7.38-7.24(m,1H),7.23-7.13(m,2H),7.02-6.94(m,2H),6.64(d,J=16.0Hz,1H),6.51(dt,J=16.0,6.8Hz,1H),5.29(dd,J=5.0,2.2Hz,1H),5.07(d,J=4.6Hz,1H),5.01(d,J=5.2Hz,1H),4.81-4.74(m,1H),4.56(t,J=5.8Hz,1H),3.69(ddd,J=11.8,5.2,2.0Hz,1H),3.57(d,J=6.4Hz,1H),3.46(dt,J=11.6,6.0Hz,1H),3.30(d,J=6.8Hz,2H),3.26-3.10(m,3H);HRMS(ESI)m / z calculated for C 21 H 24 FNNaO7[M+Na] + 456.1429,found 456.1421.

[0097] Example 6: Preparation of 2-(2-fluorophenyl)-N-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)acetamide (Compound 6)

[0098]

[0099] Adapt to change the starting compound (replace the 2-(2-fluorophenoxy)acetic acid starting material in Step 3 of Example 4 with 2-(2-fluorophenyl)acetic acid starting material), and the remaining steps are the same as in Example 4 to obtain a white solid product with a yield of 87%.1 1H NMR (400 MHz, CD3OD) δ 7.51 - 7.42 (m, 2H), 7.41 - 7.24 (m, 2H), 7.19 - 7.02 (m, 4H), 4.85 (d, J = 2.6 Hz, 1H), 3.89 (dd, J = 12.0, 2.0 Hz, 1H), 3.73 (s, 2H), 3.72 - 3.67 (m, 1H), 3.50 - 3.33 (m, 4H); HRMS (ESI) m / z calculated for C 20 H 22 FNNaO7 [M + Na] + 430.1273, found 430.1264.

[0100] Example 7: Preparation of 3-(2-fluorophenyl)-N-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)propanamide (Compound 7)

[0101]

[0102] Adapt and change the starting compound (replace the 2-(2-fluorophenoxy)acetic acid starting material in Step 3 of Example 4 with 3-(2-fluorophenyl)propanoic acid starting material), and the remaining steps are the same as in Example 4 to obtain a white solid product with a yield of 70%. 1 1H NMR (400 MHz, CD3OD) δ 7.45 - 7.36 (m, 2H), 7.29 (td, J = 7.6, 1.8 Hz, 1H), 7.22 (tdd, J = 7.4, 5.2, 1.8 Hz, 1H), 7.12 - 7.00 (m, 4H), 4.89 - 4.80 (m, 1H), 3.89 (dd, J = 12.0, 2.0 Hz, 1H), 3.69 (dd, J = 12.0, 5.2 Hz, 1H), 3.50 - 3.31 (m, 4H), 3.03 (t, J = 7.6 Hz, 2H), 2.64 (dd, J = 8.8, 6.8 Hz, 2H); HRMS (ESI) m / z calculated for C 21 H 24 FNNaO7 [M + Na] + 444.1429, found 444.1455.

[0103] Example 8: Preparation of (E)-3-(2-fluorophenyl)-N-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)phenyl)acrylamide (Compound 8)

[0104]

[0105] Adapt to the change of raw material compounds (replace the raw material of 2-(2-fluorophenoxy)acetic acid in step 3 of Example 4 with the raw material of 2-fluorocinnamic acid), and the remaining steps are the same as those in Example 4, to obtain a pale yellow solid product with a yield of 26%. 1 H NMR(400MHz,CD3OD)δ7.77(d,J=16.0Hz,1H),7.67(td,J=8.0,1.8Hz,1H),7.65-7.55(m,2H),7.44-7.39(m,1H),7.28-7.13(m,2H),7.14-7.06(m,2H),6.89(d,J=15.8Hz,1H),4.87(s,1H),3.91(dd,J=12.0,2.0Hz,1H),3.76-3.65(m,1H),3.52-3.35(m,4H);HRMS(ESI)m / zcalculated for C 21 H 22 FNNaO7[M+Na] + 442.1253,found 442.1273.

[0106] Example 9: Preparation of (E)-3-(3-hydroxy-4-methoxyphenyl)-N-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)acrylamide (Compound 9)

[0107]

[0108] Adapt to the change of raw material compounds (replace the raw material of 2-(2-fluorophenoxy)acetic acid in step 3 of Example 4 with the raw material of 3-hydroxy-4-methoxycinnamic acid), and the remaining steps are the same as those in Example 4, to obtain a pale yellow solid product with a yield of 10%. 11H NMR (400 MHz, DMSO-d6) δ 10.37 (s, 1H), 9.37 (s, 1H), 7.68 - 7.62 (m, 2H), 7.39 (d, J=15.6 Hz, 1H), 7.10 - 7.02 (m, 1H), 7.05 - 6.92 (m, 4H), 6.69 (d, J=15.6 Hz, 1H), 5.31 (d, J=4.8 Hz, 1H), 5.20 - 5.06 (m, 2H), 4.79 (d, J=7.6 Hz, 1H), 4.62 (t, J=6.0 Hz, 1H), 3.79 (s, 3H), 3.67 (dd, J=10.4, 4.8 Hz, 1H), 3.48 (dd, J=11.8, 6.0 Hz, 2H), 3.33 - 3.23 (m, 2H), 3.26 - 3.17 (m, 1H); HRMS(ESI) m / z calculated for C 22 H 25 FNaO9 [M+Na] + 470.1415, found 470.1422.

[0109] Example 10: Preparation of (E)-3-(4-hydroxyphenyl)-N-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)acrylamide (Compound 10)

[0110]

[0111] Adapt to change the starting compound (replace the 2-(2-fluorophenoxy)acetic acid starting material in Step 3 of Example 4 with 4-hydroxycinnamic acid starting material), and the remaining steps are the same as in Example 4, to obtain a white solid product with a yield of 26%. 1 1H NMR (400 MHz, DMSO-d6) δ 10.00 (s, 1H), 7.60 (d, J=8.4 Hz, 2H), 7.50 - 7.41 (m, 3H), 7.04 - 6.95 (m, 2H), 6.82 (d, J=8.4 Hz, 2H), 6.59 (d, J=15.6 Hz, 1H), 5.30 (d, J=3.6 Hz, 1H), 5.07 (d, J=28.8 Hz, 2H), 4.79 (d, J=7.4 Hz, 1H), 4.58 (s, 1H), 3.69 (d, J=11.6 Hz, 1H), 3.63 - 3.42 (m, 2H), 3.28 - 3.10 (m, 3H); HRMS(ESI) m / z calculated for C 21 H 23 NNaO8 [M+Na] +440.1313, found 440.1316.

[0112] Example 11: Preparation of (E)-3-(4-Hydroxy-3-methoxyphenyl)-N-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)acrylamide (Compound 11)

[0113]

[0114] Adapt to change the starting compound (replace the 2-(2-fluorophenoxy)acetic acid starting material in Step 3 of Example 4 with 4-hydroxy-3-methoxycinnamic acid starting material), and the remaining steps are the same as in Example 4 to obtain a pale yellow liquid product with a yield of 94%. 1 H NMR (400 MHz, DMSO-d6) δ 10.29 (s, 1H), 7.68 - 7.60 (m, 2H), 7.44 (d, J = 15.6 Hz, 1H), 7.17 (d, J = 2.0 Hz, 1H), 7.07 - 6.95 (m, 3H), 6.86 (d, J = 8.0 Hz, 1H), 6.75 (d, J = 16.0 Hz, 1H), 5.31 (t, J = 4.8 Hz, 1H), 5.17 (d, J = 4.4 Hz, 1H), 5.11 (d, J = 5.0 Hz, 1H), 4.79 (d, J = 7.6 Hz, 1H), 4.62 (t, J = 5.6 Hz, 1H), 3.81 (s, 2H), 3.72 - 3.63 (m, 1H), 3.50 - 3.44 (m, 2H), 3.28 (td, J = 8.6, 4.6 Hz, 2H), 3.26 - 3.16 (m, 1H); HRMS (ESI) m / z calculated for C 22 H 25 NNaO9 [M+Na] + 470.1399, found 470.1422.

[0115] Example 12: Preparation of N-Cinnamyl-4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)benzamide (Compound 12)

[0116]

[0117] Step 1: Synthesis of 4-(Cinnamylcarbamoyl)phenyl Acetate (Intermediate 12-2)

[0118] Weigh p-acetoxybenzoic acid (Intermediate 12-1) (1.29 g, 7.2 mmol) and (E)-3-phenylprop-2-en-1-amine (800 mg, 6.0 mmol) into a 100 mL reaction flask, add 20 mL of dichloromethane solution to dissolve, and then add HATU (2.74 g, 7.2 mmol) and triethylamine (1.17 mL, 9.0 mmol) in sequence. Stir the reaction at room temperature for 5 h. After the reaction is completed, add saturated sodium bicarbonate solution for extraction and washing. Combine the organic phases, dry over anhydrous Na2SO4, and purify by column chromatography (DCM:MeOH = 100:1 - 60:1) to obtain 1.49 g of pale yellow solid with a yield of 70%. 1 H NMR (400 MHz, DMSO-d6) δ 8.79 (t, J = 5.6 Hz, 1H), 7.98 - 7.90 (m, 2H), 7.47 - 7.36 (m, 2H), 7.32 (t, J = 7.6 Hz, 2H), 7.27 - 7.19 (m, 2H), 6.58 - 6.50 (m, 1H), 6.34 (dt, J = 16.0, 5.6 Hz, 1H), 4.07 (td, J = 5.6, 1.6 Hz, 2H), 2.29 (s, 3H).

[0119] Step 2: Synthesis of N-cinnamyl-4-hydroxybenzamide (Intermediate 12-3)

[0120] Weigh Intermediate 12-2 (1.49 g, 5.0 mmol) into a 50 mL reaction flask, add 15 mL of methanol solution to dissolve, and then slowly add sodium methoxide (540 mg, 10.0 mmol). Stir the reaction at room temperature for 1 h. After the reaction is completed, evaporate the solvent and purify by column chromatography (DCM:MeOH = 150:1 - 100:1) to obtain 896 mg of white solid with a yield of 70%. 1 H NMR (400 MHz, DMSO-d6) δ 9.96 (s, 1H), 8.49 (t, J = 5.6 Hz, 1H), 7.80 - 7.72 (m, 2H), 7.45 - 7.38 (m, 2H), 7.36 - 7.27 (m, 2H), 7.26 - 7.17 (m, 1H), 6.84 - 6.76 (m, 2H), 6.51 (d, J = 16.0 Hz, 1H), 6.32 (dt, J = 15.6, 5.6 Hz, 1H), 4.03 (t, J = 5.6 Hz, 2H).

[0121] Step 3: Synthesis of N-cinnamyl-4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)benzamide (Compound 12)

[0122] Weigh the intermediate 12-3 (800 mg, 2.7 mmol), (3S,4R,5R,6S)-6-(acetyloxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetraacetate (1.27 g, 3.3 mmol), Molecular sieve (3.00 g) into a 50 mL reaction flask, add 12 mL of dichloromethane solution to dissolve, slowly add boron trifluoride ether complex (1.03 mL, 8.1 mmol) dropwise under ice bath, stir at room temperature for 24 h. After the reaction is completed, add 1 M sodium hydroxide solution for extraction, combine the organic phases, dry over anhydrous Na2SO4, evaporate the solvent, then add 10 mL of methanol solution to dissolve, and slowly add sodium methoxide (292 mg, 5.4 mmol), stir at room temperature for 1 h. After the reaction is completed, evaporate the solvent and purify by column chromatography (DCM:MeOH = 40:1 - 5:1) to obtain 135 mg of white solid, with a yield of 12%. 1 1H NMR (400 MHz, CD3OD) δ 8.50 (s, 1H), 7.88 - 7.81 (m, 2H), 7.43 - 7.36 (m, 2H), 7.29 (dd, J = 8.4, 6.8 Hz, 2H), 7.19 - 7.12 (m, 2H), 6.59 (d, J = 16.0 Hz, 1H), 6.33 (dt, J = 15.6, 6.0 Hz, 1H), 5.01 (s, 1H), 4.14 (dd, J = 6.0, 1.6 Hz, 2H), 3.90 (dd, J = 12.1, 2.2 Hz, 1H), 3.70 (dd, J = 12.4, 5.6 Hz, 1H), 3.51 - 3.46 (m, 3H), 3.43 - 3.37 (m, 1H); HRMS (ESI) m / z calculated for C 22 H 25 NNaO7 [M+Na] + 438.1523, found 438.1541.

[0123] Example 13: Preparation of N-phenethyl-4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)benzamide (Compound 13)

[0124]

[0125] Adapt to change the starting compound (replace the (E)-3-phenylprop-2-en-1-amine starting material in Step 1 of Example 12 with 2-phenethyl-1-amine starting material), and the remaining steps are the same as in Example 12 to obtain a white solid product with a yield of 50%. 11H NMR (400 MHz, DMSO-d6) δ 8.45 (t, J = 5.6 Hz, 1H), 7.78 (d, J = 8.4 Hz, 2H), 7.29 (t, J = 7.4 Hz, 2H), 7.27 - 7.15 (m, 3H), 7.06 (d, J = 8.4 Hz, 2H), 5.37 (s, 1H), 5.14 (d, J = 22 Hz, 2H), 4.95 (d, J = 7.2 Hz, 1H), 4.59 (t, J = 5.6 Hz, 1H), 3.69 (dd, J = 11.6, 4.6 Hz, 1H), 3.50 - 3.38 (m, 4H), 3.31 - 3.22 (m, 2H), 3.16 (t, J = 9.0 Hz, 1H), 2.83 (t, J = 7.4 Hz, 2H); HRMS (ESI) m / z calculated for C 21 H 25 NNaO7 [M+Na] + 426.1523, found 426.1504.

[0126] Example 14: Preparation of N-(2-fluorobenzyl)-4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)benzamide (Compound 14)

[0127]

[0128] Adapt to change the starting compound (replace the (E)-3-phenylprop-2-en-1-amine starting material in Step 1 of Example 12 with 2-fluorobenzylamine starting material), and the remaining steps are the same as in Example 12 to obtain a white solid product with a yield of 50%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.91 (s, 1H), 7.99 - 7.74 (m, 2H), 7.32 (d, J = 20.4 Hz, 2H), 7.22 - 7.02 (m, 4H), 5.36 (s, 1H), 5.17 - 4.92 (m, 3H), 4.53 (d, J = 28.2 Hz, 3H), 3.69 (d, J = 12.0 Hz, 1H), 3.55 - 3.41 (m, 1H), 3.22 (d, J = 37.6 Hz, 4H); HRMS (ESI) m / z calculated for C 20 H 22 FNNaO7 [M+Na] + 430.1273, found 430.1270.

[0129] Example 15: Preparation of N-(2-fluorophenethyl)-4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)benzamide (Compound 15)

[0130]

[0131] Adapt to change the starting compound (replace the (E)-3-phenylprop-2-en-1-amine starting material in Step 1 of Example 12 with 2-(2-fluorophenyl)ethan-1-amine starting material), and the remaining steps are the same as in Example 12 to obtain a white solid product with a yield of 52%. 1 HNMR(400MHz,CD3OD)δ7.82-7.67(m,2H),7.32-7.18(m,2H),7.18-6.98(m,4H),5.04-4.94(m,1H),3.90(dd,J=12.2,2.2Hz,1H),3.70(dd,J=12.0,5.6Hz,1H),3.59(t,J=7.2Hz,2H),3.48(qd,J=6.4,2.8Hz,3H),3.41-3.37(m,1H),2.96(t,J=7.2Hz,2H);HRMS(ESI)m / zcalculated for C 21 H 24 FNNaO7[M+Na] + 444.1429,found 444.1425.

[0132] Example 16: Preparation of N'-((E)-2-fluorobenzylidene)-4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)benzohydrazide (Compound 16), and its synthetic route is as follows:

[0133]

[0134] Step 1: Synthesis of (E)-N'-(2-fluorobenzylidene)-4-hydroxybenzohydrazide (Intermediate 16-2)

[0135] Weigh p-hydroxybenzohydrazide (Intermediate 16-1) (800 mg, 5.3 mmol) into a 50 mL reaction flask, add 15 mL of ethanol solution to dissolve it, and slowly add 2-fluorobenzaldehyde (0.65 mL, 6.3 mmol). React under reflux at 80 °C for 1 h. After the reaction is completed, evaporate the solvent and purify by column chromatography (DCM:MeOH = 150:1 to 80:1) to obtain 1.28 g of a white solid with a yield of 94%. 11H NMR (400 MHz, DMSO-d6) δ 11.78 (s, 1H), 10.15 (s, 1H), 8.68 (s, 1H), 7.93 (d, J = 7.8 Hz, 1H), 7.82 (d, J = 8.4 Hz, 2H), 7.51 - 7.45 (m, 1H), 7.34 - 7.25 (m, 2H), 6.90 - 6.83 (m, 2H).

[0136] Step 2: Synthesis of N'-((E)-2-fluorobenzylidene)-4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)benzohydrazide (Compound 16)

[0137] Adapt and change the starting material compound (replace the intermediate N-cinnamyl-4-hydroxybenzamide starting material in Step 3 of Example 12 with the intermediate (E)-N'-(2-fluorobenzylidene)-4-hydroxybenzohydrazide starting material (e.g., replace the compound with the structural formula 12-3 with the structural formula 16-2)), and the remaining steps are the same as those in Step 3 of Example 12, to obtain a white solid product with a yield of 49%. 1 1H NMR (400 MHz, CD3OD) δ 8.63 (s, 1H), 8.22 (td, J = 7.6, 1.6 Hz, 1H), 7.97 - 7.91 (m, 2H), 7.51 - 7.41 (m, 1H), 7.30 - 7.13 (m, 4H), 5.07 - 4.99 (m, 1H), 3.92 (dd, J = 12.0, 2.2 Hz, 1H), 3.71 (dd, J = 12.0, 5.6 Hz, 1H), 3.55 - 3.46 (m, 3H), 3.43 - 3.38 (m, 1H); HRMS (ESI) m / z calculated for C 20 H 21 FN2NaO7 [M + Na] + 443.1225, found 443.1224.

[0138] Example 17: Preparation of (E)-3-(2-fluorophenyl)-1-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)propan-2-one (Compound 17), and its synthetic route is as follows:

[0139]

[0140] Step 1: Synthesis of (E)-3-(2-fluorophenyl)-1-(4-hydroxyphenyl)prop-2-en-1-one (Intermediate 17-2)

[0141] Weigh p - hydroxyacetophenone (2.00 g, 14.7 mmol) (Intermediate 17 - 1) into a 100 mL reaction flask, add 15 mL of ethanol solution to dissolve it, and slowly drip the ethanol solution of 2 - fluorobenzaldehyde (2.16 g, 17.7 mmol). Stir the reaction at room temperature overnight. After the reaction is completed, evaporate part of the solvent, adjust the pH to 2 with 2M hydrochloric acid solution, precipitate the solid, filter it by suction, wash it with water, and dry it to obtain 3.10 g of light yellow solid, with a yield of 87%. 1 H NMR (400 MHz, DMSO - d6) δ 10.47 (s, 1H), 8.15 - 8.02 (m, 3H), 7.96 (d, J = 15.6 Hz, 1H), 7.78 (d, J = 15.6 Hz, 1H), 7.53 - 7.48 (m, 1H), 7.36 - 7.26 (m, 2H), 6.95 - 6.86 (m, 2H).

[0142] Step 2: Synthesis of (E) - 3 - (2 - fluorophenyl) - 1 - (4 - ((2S,3R,4S,5S,6R) - 3,4,5 - trihydroxy - 6 - (hydroxymethyl)tetrahydro - 2H - pyran - 2 - yl)oxy)phenyl)prop - 2 - en - 1 - one (Compound 17)

[0143] Adapt to change the starting compound, and its preparation and synthesis method are similar to Step 3 of Compound 12. It is a light yellow solid with a yield of 22%. 1 H NMR (400 MHz, DMSO - d6) δ 8.16 (d, J = 9.2 Hz, 2H), 8.01 (d, J = 15.6 Hz, 1H), 7.82 (d, J = 15.6 Hz, 1H), 7.55 - 7.49 (m, 1H), 7.38 - 7.27 (m, 2H), 7.22 - 7.07 (m, 3H), 5.40 (d, J = 4.8 Hz, 1H), 5.14 (d, J = 4.6 Hz, 1H), 5.06 (t, J = 5.2 Hz, 2H), 4.60 (t, J = 5.6 Hz, 1H), 3.73 - 3.68 (m, 1H), 3.51 - 3.39 (m, 2H), 3.31 - 3.25 (m, 2H), 3.22 - 3.18 (m, 1H); HRMS (ESI) m / z calculated for C 21 H 21 FNNaO7[M + Na] + 427.1184, found 427.1180.

[0144] Example 18: Preparation of N-(2-fluorophenyl)-2-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)thio)acetamide (Compound 18), and its synthetic route is as follows:

[0145]

[0146] Step 1: Synthesis of N-(2-fluorophenyl)-2-((4-methoxyphenyl)thio)acetamide (Intermediate 18-2)

[0147] Weigh 2-bromo-N-(2-fluorophenyl)acetamide (Intermediate 18-1) (500 mg, 2.2 mmol), p-methoxythiophenol (362 mg, 2.6 mmol), cesium carbonate (1.26 g, 3.9 mmol), and potassium iodide (36 mg, 0.2 mmol) into a 50 mL reaction flask, add 12 mL of acetonitrile solution to dissolve, and reflux for 48 h. After the reaction is completed, evaporate the solvent, extract with ethyl acetate - water, combine the organic phases, dry over anhydrous Na2SO4, and purify by column chromatography (PE:EA = 20:1 - 2:1) to obtain 384 mg of a pale yellow solid with a yield of 61%. 1 H NMR (400 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.86 - 7.79 (m, 1H), 7.43 - 7.37 (m, 3H), 7.18 - 7.13 (m, 3H), 6.95 - 6.88 (m, 2H), 3.76 (d, J = 2.4 Hz, 3H), 3.73 (s, 2H).

[0148] Step 2: Synthesis of N-(2-fluorophenyl)-2-((4-hydroxyphenyl)thio)acetamide (Intermediate 18-3)

[0149] Weigh Intermediate 18-2 (350 mg, 1.2 mmol) into a 50 mL reaction flask, add 10 mL of dichloromethane solution to dissolve, and slowly add 2 M boron tribromide dichloromethane solution (1.20 mL, 2.4 mmol) dropwise under ice bath, and stir at room temperature for 72 h. After the reaction is completed, quench with methanol solution, wash with saturated sodium bicarbonate solution, combine the organic phases, dry over anhydrous Na2SO4, and purify by column chromatography (DCM:MeOH = 120:1 - 60:1) to obtain 66 mg of a pale yellow liquid with a yield of 20%. 11H NMR (400 MHz, DMSO-d6) δ 9.93 (s, 1H), 8.32 (s, 1H), 7.63 (d, J = 8.8 Hz, 1H), 7.46 - 7.43 (m, 2H), 7.16 (t, J = 8.2 Hz, 2H), 7.04 (d, J = 8.8 Hz, 1H), 6.96 - 6.92 (m, 2H), 3.76 (s, 2H).

[0150] Step 3: Synthesis of N-(2-Fluorophenyl)-2-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)thio)acetamide (Compound 18)

[0151] Adapt to change the starting compound (replace the intermediate N-cinnamyl-4-hydroxybenzamide starting material in Step 3 of Example 12 with the intermediate N-(2-fluorophenyl)-2-((4-hydroxyphenyl)thio)acetamide starting material (e.g., replace the compound with the structural formula 12-3 with the structural formula 18-3)), and the remaining steps are the same as those in Step 3 of Example 12, to obtain a pale yellow liquid product with a yield of 50%. 1 1H NMR (400 MHz, CD3OD) δ 7.85 - 7.76 (m, 1H), 7.50 - 7.42 (m, 2H), 7.16 - 7.11 (m, 3H), 7.10 - 7.03 (m, 2H), 4.92 (d, J = 10.6 Hz, 1H), 3.88 (dt, J = 12.0, 3.8 Hz, 1H), 3.75 - 3.65 (m, 3H), 3.53 - 3.37 (m, 4H); HRMS (ESI) m / z calculated for C 20 H 22 FNNaO7S [M+Na] + 462.0993, found 462.0977.

[0152] Example 19: Preparation of N-(2-Fluorophenyl)-2-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenoxy)acetamide (Compound 19)

[0153]

[0154] Adapt to change the starting compound (replace the N-(2-fluorophenyl)-2-((4-hydroxyphenyl)thio)acetamide starting material in Step 3 of Example 18 with the N-(2-fluorophenyl)-2-(4-hydroxyphenoxy)acetamide starting material), and the remaining steps are the same as those in Example 18, to obtain a white solid product with a yield of 99%. 11H NMR (400 MHz, CD3OD) δ 7.96 - 7.87 (m, 1H), 7.24 - 7.12 (m, 3H), 7.14 - 7.06 (m, 2H), 7.02 - 6.95 (m, 2H), 4.84 - 4.78 (m, 1H), 4.67 (s, 2H), 3.89 (dd, J=12.0, 1.8 Hz, 1H), 3.70 (dd, J=12.0, 4.8 Hz, 1H), 3.49 - 3.36 (m, 4H); HRMS (ESI) m / z calculated for C 20 H 22 FNNaO8 [M+Na] + 446.1222, found 446.1232.

[0155] Example 20: Preparation of (E)-N-(2-fluorophenyl)-3-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenoxy)acrylamide (Compound 20), and its synthetic route is as follows:

[0156]

[0157] Step 1: Synthesis of (E)-3-(4-methoxyphenoxy)acrylic acid (Intermediate 20-2)

[0158] Weigh 4-methoxyphenol (Intermediate 20-1) (2.00 g, 16.1 mmol) and 1,4-diazabicyclo[2,2,2]octane (DABCO) (181 g, 1.6 mmol) into a 100 mL reaction flask, add 20 mL of tetrahydrofuran solution to dissolve, slowly add methyl propiolate (1.31 mL, 14.7 mmol) dropwise under ice bath, and then stir the reaction at room temperature for 8 h. After the reaction is completed, concentrate, wash with ethyl acetate and 1 M NaOH solution, combine the organic phases, dry over anhydrous Na2SO4, concentrate and dissolve in 30 mL of tetrahydrofuran-distilled water solution (1:1), slowly add lithium hydroxide (1.85 g, 44.0 mmol), and stir the reaction at room temperature for 12 h. After the reaction is completed, evaporate the solvent, dissolve in dichloromethane, wash twice with 1 M NaOH solution, adjust the pH of the aqueous phase to 3 with 3 M hydrochloric acid solution, precipitate a solid, filter by suction, wash with water, and dry to obtain 3.10 g of a white solid with a yield of 99%. 1 1H NMR (400 MHz, DMSO-d6) δ 12.05 (s, 1H), 7.69 (d, J=12.2 Hz, 1H), 7.15 - 7.10 (m, 2H), 6.98 - 6.95 (m, 2H), 5.35 (d, J=12.2 Hz, 1H), 3.75 (s, 3H).

[0159] Step 2: Synthesis of (E)-N-(2-fluorophenyl)-3-(4-hydroxyphenoxy)acrylamide (Intermediate 20-3)

[0160] Weigh intermediate 20-2 (1.49 g, 7.7 mmol) and 2-fluoroaniline (0.66 mL, 7.7 mmol) into a 50 mL reaction flask, add 20 mL of dichloromethane solution to dissolve, and successively add HATU (3.50 mg, 9.2 mmol) and triethylamine (1.49 mL, 11.5 mmol), then stir and react at room temperature for 5 h. After the reaction is completed, extract and wash successively with saturated sodium bicarbonate solution, 1 M hydrochloric acid solution, and saturated brine. The organic phase is dried over anhydrous Na2SO4 and then concentrated. Add 15 mL of dichloromethane solution to dissolve, and slowly dropwise add a dichloromethane solution of 2 M boron tribromide (11.55 mL, 23.1 mmol) under ice bath, and stir and react at room temperature for 72 h. After the reaction is completed, quench with methanol solution, extract and wash with saturated sodium bicarbonate solution, combine the organic phases, dry over anhydrous Na2SO4, and purify by column chromatography (DCM:MeOH = 120:1 - 60:1) to obtain 630 mg of white solid with a yield of 30%. 1 H NMR (400 MHz, CDCl3) δ 8.47 - 8.41 (m, 1H), 7.82 (d, J = 11.6 Hz, 1H), 7.45 (dd, J = 8.4, 4.4 Hz, 1H), 7.17 - 7.02 (m, 3H), 6.99 - 6.95 (m, 2H), 6.87 - 6.84 (m, 2H), 5.72 (d, J = 12.0 Hz, 1H).

[0161] Step 3: Synthesis of (E)-N-(2-fluorophenyl)-3-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenoxy)acrylamide (Compound 20)

[0162] Adapt and change the starting compound (replace the starting intermediate N-cinnamyl-4-hydroxybenzamide in Step 3 of Example 12 with the intermediate (E)-N-(2-fluorophenyl)-3-(4-hydroxyphenoxy)acrylamide (for example, replace the compound with the structural formula 12-3 with the structural formula 20-3)), and the remaining steps are the same as Step 3 of Example 12 to obtain a pale yellow liquid product with a yield of 42%. 11H NMR (400 MHz, CD3OD) δ 7.92 (d, J = 9.4 Hz, 1H), 7.81 (dd, J = 12.4, 1.8 Hz, 1H), 7.19 - 7.12 (m, 4H), 7.11 - 7.03 (m, 2H), 5.94 (d, J = 12.0 Hz, 1H), 5.45 (d, J = 12.2 Hz, 1H), 3.90 (dt, J = 12.0, 2.0 Hz, 1H), 3.75 - 3.66 (m, 2H), 3.61 (s, 1H), 3.48 - 3.43 (m, 3H); HRMS (ESI) m / z calculated for C 21 H 22 FNNaO8 [M+Na] + 458.1222, found 458.1231.

[0163] Example 21: Preparation of (2S,3R,4S,5S,6R)-2-(4-(1-cinnamoyl-1H-1,2,3-triazol-4-yl)phenoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (Compound 21), and its synthetic route is as follows:

[0164]

[0165] Step 1: Synthesis of 1-cinnamoyl-4-(4-methoxyphenyl)-1H-1,2,3-triazole (Intermediate 21-2)

[0166] Weigh (E)-(3-azidoprop-1-en-1-yl)benzene (Intermediate 21-1) (1.00 g, 6.3 mmol) and 4-ethynylanisole (1.66 g, 12.6 mmol) into a 100 mL reaction flask, add 50 mL of tert-butanol-distilled water (1:1) solution to dissolve, slowly add sodium ascorbate (3.07 g, 16.3 mmol) and copper sulfate pentahydrate (3.15 g, 12.6 mmol), and stir the reaction at room temperature for 20 h. After the reaction is completed, extract with ethyl acetate-water, dry over anhydrous Na2SO4, and purify by column chromatography (DCM:MeOH = 150:1 - 100:1) to obtain 1.47 g of a gray solid with a yield of 80%. 11H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1H), 7.87 - 7.76 (m, 2H), 7.52 - 7.46 (m, 2H), 7.39 - 7.33 (m, 2H), 7.31 - 7.25 (m, 1H), 7.04 - 6.97 (m, 2H), 6.71 (d, J = 16.0 Hz, 1H), 6.55 (dt, J = 16.0, 6.4 Hz, 1H), 5.20 (dd, J = 6.4, 1.4 Hz, 2H), 3.78 (s, 3H).

[0167] Step 2: Synthesis of 4-(1-cinnamoyl-1H-1,2,3-triazol-4-yl)phenol (Intermediate 21-3)

[0168] Weigh Intermediate 21-2 (1.47 g, 5.0 mmol) into a 50 mL reaction flask, add 12 mL of dichloromethane solution to dissolve it, and slowly add a dichloromethane solution of 2 M boron tribromide (7.50 mL, 15.0 mmol) dropwise under an ice bath. Stir the reaction at room temperature for 72 h. After the reaction is completed, quench it with methanol solution, wash it with saturated sodium bicarbonate solution, combine the organic phases, dry over anhydrous Na2SO4, and purify by column chromatography (DCM:MeOH = 120:1 - 60:1) to obtain 690 mg of a yellow solid with a yield of 50%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.43 (s, 1H), 7.69 - 7.60 (m, 2H), 7.51 - 7.45 (m, 2H), 7.38 - 7.32 (m, 2H), 7.31 - 7.24 (m, 1H), 6.85 - 6.78 (m, 2H), 6.70 (d, J = 15.6 Hz, 1H), 6.54 (dt, J = 16.0, 6.4 Hz, 1H), 5.19 (dd, J = 6.4, 1.2 Hz, 2H).

[0169] Step 3: Synthesis of (2S,3R,4S,5S,6R)-2-(4-(1-cinnamoyl-1H-1,2,3-triazol-4-yl)phenoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (Compound 21)

[0170] Adapt and change the starting compounds (replace the starting intermediate N-cinnamyl-4-hydroxybenzamide in Step 3 of Example 12 with the starting intermediate 4-(1-cinnamoyl-1H-1,2,3-triazol-4-yl)phenol (for example, replace the compound with the structural formula 12-3 with the compound with the structural formula 21-3)), and the remaining steps are the same as those in Step 3 of Example 12 to obtain a white solid product with a yield of 19%. 1HNMR(400MHz,CD3OD)δ8.30(s,1H),7.77(d,J=8.4Hz,2H),7.46(d,J=7.6Hz,2H),7.29(dt,J=26.0,7.2Hz,3H),7.17(d,J=8.4Hz,2H),6.76(d,J=15.8Hz,1H),6.50(dt,J=15.4,6.6Hz,1H),5.23(d,J=6.6Hz,1H),3.94-3.87(m,1H),3.71(dd,J=12.2,5.4Hz,1H),3.60(s,1H),3.49-3.45(m,3H);HRMS(ESI)m / z calculated for C 23 H 25 N3NaO6[M+Na] + 462.1636,found462.1651.

[0171] Example22:Preparation of (E)-3-(2-fluorophenyl)-N-(4'-hydroxy-[1,1'-biphenyl]-4-yl)acrylamide (Compound 22), and its synthetic route is as follows:

[0172]

[0173] Step 1:Synthesis of 4-(cinnamoylamino-carbonyl)phenyl acetate (Intermediate 22-2)

[0174] Weigh o-fluorocinnamic acid (Intermediate 22-1) (1.00 g, 6.0 mmol) and p-bromoaniline (1.04 mg, 6.0 mmol) into a 50 mL reaction flask, add 15 mL of dichloromethane solution to dissolve, and then add HATU (2.75 g, 7.2 mmol) and triethylamine (1.17 mL, 9.0 mmol) in sequence, and stir the reaction at room temperature for 5 h. After the reaction is completed, add saturated sodium bicarbonate solution for extraction and washing, combine the organic phases, dry over anhydrous Na2SO4, and purify by column chromatography (DCM:MeOH = 150:1 to 80:1) to obtain 1.19 g of white solid with a yield of 62%. 1 H NMR(400MHz,DMSO-d6)δ10.44(s,1H),7.77-7.67(m,3H),7.64(d,J=16.0Hz,1H),7.56-7.42(m,3H),7.37-7.26(m,2H),6.92(d,J=16.0Hz,1H).

[0175] Step 2:Synthesis of (E)-3-(2-fluorophenyl)-N-(4'-hydroxy-[1,1'-biphenyl]-4-yl)acrylamide (Compound 22)

[0176] Weigh the intermediate 22-2 (500 mg, 1.6 mmol), 4-hydroxyphenylboronic acid pinacol ester (516 mg, 2.3 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride dichloromethane complex (Pd(dppf)2Cl2·CH2Cl2) (255 mg, 0.3 mmol), and potassium phosphate (829 mg, 3.9 mmol) into a 50 mL reaction flask. Add 15 mL of a dioxane-distilled water (4:1) solution to dissolve them. After protecting with argon, reflux the reaction for 15 h. After the reaction is completed, evaporate the solvent and purify by column chromatography (DCM:MeOH = 180:1 - 120:1) to obtain 234 mg of a yellow solid with a yield of 45%. 1 H NMR (400 MHz, DMSO-d6) δ 10.35 (s, 1H), 9.51 (s, 1H), 7.78 - 7.69 (m, 3H), 7.65 (d, J = 15.8 Hz, 1H), 7.58 - 7.54 (m, 2H), 7.50 - 7.44 (m, 3H), 7.35 - 7.27 (m, 2H), 6.97 (d, J = 15.8 Hz, 1H), 6.87 - 6.80 (m, 2H). HRMS (ESI) m / z calculated for C 23 H 25 N3NaO6 [M+Na] + 462.1636, found 462.1651. HRMS (ESI) m / z calculated for C 21 H 16 FNNaO2 [M+Na] + 356.1057, found 356.1066.

[0177] Example 23: Preparation of (E)-3-(2-fluorophenyl)-N-(5-(4-hydroxyphenyl)pyridin-2-yl)acrylamide (Compound 23)

[0178]

[0179] Adapt and change the starting compound (replace the p-bromoaniline starting material in Step 1 of Example 22 with a 5-bromopyridin-2-amine starting material), and the remaining steps are the same as in Example 22 to obtain a brown solid product with a yield of 46%. 11H NMR (400 MHz, DMSO-d6) δ 10.87 (s, 1H), 9.63 (s, 1H), 8.60 (dd, J = 2.4, 0.8 Hz, 1H), 8.29 (d, J = 8.6 Hz, 1H), 8.04 (dd, J = 8.8, 2.6 Hz, 1H), 7.75 - 7.66 (m, 2H), 7.58 - 7.52 (m, 2H), 7.51 - 7.45 (m, 1H), 7.36 - 7.27 (m, 2H), 7.16 (d, J = 15.6 Hz, 1H), 6.90 - 6.83 (m, 2H). HRMS (ESI) m / z calculated for C 20 H 15 FN2NaO2 [M+Na] + 357.1010, found 357.1012.

[0180] Example 24: Preparation of 2-(2-fluorophenoxy)-N-(4'-hydroxy-[1,1'-biphenyl]-4-yl)acetamide (Compound 24)

[0181]

[0182] Adapt to change the starting compound (replace the starting material of o-fluorocinnamic acid in Step 1 of Example 22 with 2-(2-fluorophenoxy)acetic acid), and the remaining steps are the same as in Example 22 to obtain a brown solid product with a yield of 73%. 1 1H NMR (400 MHz, DMSO-d6) δ 10.20 (s, 1H), 9.50 (s, 1H), 7.69 - 7.62 (m, 2H), 7.57 - 7.42 (m, 4H), 7.30 - 7.20 (m, 1H), 7.13 (dd, J = 5.6, 3.0 Hz, 2H), 7.03 - 6.93 (m, 1H), 6.86 - 6.79 (m, 2H), 4.81 (s, 2H). HRMS (ESI) m / z calculated for C 20 H 16 FNNaO3 [M+Na] + 360.1006, found 360.1015.

[0183] Example 25: Preparation of 2-(2-fluorophenoxy)-N-(5-(4-hydroxyphenyl)pyridin-2-yl)acetamide (Compound 25)

[0184]

[0185] Adapt to change the raw material compounds (replace the p-bromoaniline raw material in step 1 of Example 22 with 5-bromopyridin-2-amine raw material, and replace the o-fluorocinnamic acid raw material with 2-(2-fluorophenoxy)acetic acid raw material), and the remaining steps are the same as those in Example 22 to obtain a pale yellow solid product with a yield of 41%. 1 H NMR(400MHz,DMSO-d6)δ10.61(s,1H),9.63(s,1H),8.58(d,J=2.8Hz,1H),8.12-7.98(m,2H),7.56-7.50(m,2H),7.30-7.20(m,1H),7.17-7.07(m,2H),7.03-6.92(m,1H),6.89-6.82(m,2H),4.90(s,2H).HRMS(ESI)m / z calculated forC 19 H 15 FN2NaO3[M+Na] + 361.0959,found 361.0973.

[0186] Example 26: Preparation of N-(2-fluorophenyl)-2-((4'-hydroxy-[1,1'-biphenyl]-4-yl)oxy)acetamide (Compound 26)

[0187]

[0188] Adapt to change the raw material compounds (replace the (E)-3-(2-fluorophenyl)-N-(4'-hydroxy-[1,1'-biphenyl]-4-yl)acrylamide raw material in step 2 of Example 22 with 2-(4-bromophenoxy)-N-(2-fluorophenyl)acetamide raw material), and the remaining steps are the same as those in Example 22 to obtain a white solid product with a yield of 47%. 1 H NMR(400MHz,DMSO-d6)δ9.90(s,1H),9.46(s,1H),7.83(td,J=7.6,3.6Hz,1H),7.55-7.49(m,2H),7.46-7.39(m,2H),7.33-7.14(m,3H),7.07-7.00(m,2H),6.85-6.78(m,2H),4.79(s,2H).HRMS(ESI)m / z calculated forC 20 H 16 FNNaO3[M+Na] + 360.1006,found 360.1007.

[0189] Example 27: Preparation of N-(2-fluorophenyl)-2-((5-(4-hydroxyphenyl)pyridin-2-yl)oxy)acetamide (Compound 27)

[0190]

[0191] Adapt to change the starting compound (replace the (E)-3-(2-fluorophenyl)-N-(4'-hydroxy-[1,1'-biphenyl]-4-yl)acrylamide starting material in Step 2 of Example 22 with 2-((5-bromopyridin-2-yl)oxy)-N-(2-fluorophenyl)acetamide starting material), and the remaining steps are the same as in Example 22 to obtain a white solid product with a yield of 44%. 1 H NMR(400MHz,DMSO-d6)δ10.19(s,1H),9.51(s,1H),7.99-7.89(m,2H),7.79(dd,J=9.6,2.8Hz,1H),7.39-7.33(m,2H),7.33-7.23(m,1H),7.15(dd,J=6.6,3.2Hz,2H),6.89-6.78(m,2H),6.47(d,J=9.2Hz,1H),4.88(s,2H).HRMS(ESI)m / z calculated for C 20 H 15 FN2NaO2[M+Na] + 357.1010,found357.1012.HRMS(ESI)m / z calculated for C 19 H 15 FN2NaO3[M+Na] + 361.0959,found361.0969.

[0192] Example 28: Preparation of 3-(2-fluorophenyl)-N-(5-(4-hydroxyphenyl)pyridin-2-yl)propanamide (Compound 28)

[0193]

[0194] Adapt to change the starting compound (replace the 4-bromoaniline starting material in Step 1 of Example 22 with 5-bromopyridin-2-amine starting material, and the o-fluorocinnamic acid starting material with 3-(2-fluorophenoxy)propanoic acid starting material), and the remaining steps are the same as in Example 22 to obtain a brownish-yellow liquid product with a yield of 99%. 11H NMR (400 MHz, DMSO-d6) δ 10.54 (s, 1H), 9.59 (s, 1H), 8.53 (dd, J = 2.4, 0.8 Hz, 1H), 8.11 (d, J = 8.8 Hz, 1H), 7.97 (dd, J = 8.4, 2.6 Hz, 1H), 7.54 - 7.48 (m, 2H), 7.33 (td, J = 7.6, 1.8 Hz, 1H), 7.25 (tdd, J = 7.2, 5.4, 1.8 Hz, 1H), 7.19 - 7.08 (m, 2H), 6.88 - 6.82 (m, 2H), 2.98 - 2.86 (m, 2H), 2.72 (dd, J = 8.8, 5.6 Hz, 2H). HRMS (ESI) m / z calculated for C 20 H 18 FN2O2 [M+H] + 337.1347, found 337.1344.

[0195] Example 29: Preparation of 3-(2-fluorophenyl)-N-(5-(4-(2-hydroxyethyl)phenyl)pyridin-2-yl)propanamide (Compound 29)

[0196]

[0197] Adapt to change the raw materials (replace the raw material of 4-hydroxyphenylboronic acid pinacol ester in Step 2 of Example 22 with 2-[4-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethan-1-ol), and the synthesis method is the same as that of Example 22, to obtain a pale yellow solid product with a yield of 40%. 1 1H NMR (400 MHz, DMSO-d6) δ 10.60 (s, 1H), 8.60 (d, J = 2.4 Hz, 1H), 8.15 (d, J = 8.8 Hz, 1H), 8.05 (dd, J = 8.8, 2.4 Hz, 1H), 7.65 - 7.55 (m, 2H), 7.36 - 7.31 (m, 3H), 7.30 - 7.20 (m, 1H), 7.22 - 7.08 (m, 2H), 4.67 (t, J = 5.2 Hz, 1H), 3.62 (td, J = 7.0, 5.2 Hz, 2H), 2.94 (t, J = 7.6 Hz, 2H), 2.80 - 2.69 (m, 4H). HRMS (ESI) m / z calculated for C 22 H 22 FN2O2 [M+H] + 365.1660, found 365.1660.

[0198] Example 30: Preparation of (E)-3-(2-fluorophenyl)-N-(4'-(2-hydroxyethyl)-[1,1'-biphenyl]-4-yl)acrylamide (Compound 30)

[0199]

[0200] Adapt to change the starting compound (replace the 4-hydroxyphenylboronic acid pinacol ester starting material in Step 2 of Example 22 with 2-[4-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethan-1-ol). The synthesis method is the same as that of Example 22 to obtain a white solid product with a yield of 40%. 1 H NMR(400MHz,DMSO-d6)δ10.40(s,1H),7.83-7.76(m,2H),7.73(td,J=7.8,1.6Hz,1H),7.70-7.60(m,2H),7.61-7.53(m,2H),7.50-7.45(m,1H),7.37-7.26(m,4H),6.97(d,J=15.8Hz,1H),4.66(t,J=5.2Hz,1H),3.63(td,J=7.0,5.2Hz,2H),2.75(t,J=7.0Hz,2H).HRMS(ESI)m / z calculated for C 23 H 21 FNO2[M+H] + 362.1551,found362.1550.

[0201] Example 31: Preparation of (E)-3-(2-fluorophenyl)-N-(5-(4-(2-hydroxyethyl)phenyl)pyridin-2-yl)acrylamide (Compound 31)

[0202]

[0203] Adapt to change the starting compounds (replace the p-bromoaniline starting material in Step 1 of Example 22 with 5-bromopyridin-2-amine starting material, and replace the 4-hydroxyphenylboronic acid pinacol ester starting material with 2-[4-(tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethan-1-ol). The remaining steps are the same as those of Example 22 to obtain a white solid product with a yield of 18%. 11H NMR (400 MHz, DMSO-d6) δ 10.92 (s, 1H), 8.66 (dd, J = 2.4, 0.8 Hz, 1H), 8.36 - 8.29 (m, 1H), 8.11 (dd, J = 8.8, 2.8 Hz, 1H), 7.76 - 7.60 (m, 4H), 7.51 - 7.46 (m, 1H), 7.36 - 7.27 (m, 4H), 7.17 (d, J = 15.6 Hz, 1H), 4.67 (t, J = 5.2 Hz, 1H), 3.63 (td, J = 7.0, 5.2 Hz, 2H), 2.77 (t, J = 7.0 Hz, 2H). HRMS (ESI) m / z calculated for C 22 H 20 FN2O2 [M + H] + 363.1503, found 363.1514.

[0204] Example 32: Preparation of (E)-3-(2-fluorophenyl)-N-(4'-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-[1,1'-biphenyl]-4-yl)acrylamide (Compound 32), and its synthetic route is as follows:

[0205]

[0206] Step 1: Synthesis of (2R,3R,4S,5R,6R)-2-(acetoxymethyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triacetate (Intermediate 32-2)

[0207] Weigh (3S,4R,5R,6S)-6-(acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetraacetate (Intermediate 32-1) (1.00 g, 2.6 mmol) into a 50 mL reaction flask, add 15 mL of tetrahydrofuran solution to dissolve it, slowly add benzylamine (0.31 mL, 2.8 mmol), and stir the reaction at room temperature for 24 h. After the reaction is completed, evaporate the solvent, and purify by column chromatography (PE:EA = 15:1 - 2:1) to obtain 1.10 g of a pale yellow liquid with a yield of 99%. 1 1H NMR (400 MHz, DMSO-d6) δ 7.30 (t, J = 11.6 Hz, 1H), 5.35 (t, J = 9.8 Hz, 1H), 5.22 (q, J = 4.6, 4.0 Hz, 1H), 4.94 - 4.83 (m, 1H), 4.70 (dd, J = 11.6, 8.0 Hz, 1H), 4.13 (d, J = 9.4 Hz, 2H), 4.04 - 3.96 (m, 2H), 2.03 - 1.90 (m, 12H).

[0208] Step 2: Synthesis of (2R,3R,4S,5R,6S)-2-(acetoxymethyl)-6-(2,2,2-trichloro-1-iminoethoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (Intermediate 32-3)

[0209] Weigh intermediate 32-2 (1.10 g, 3.2 mmol) into a 50 mL reaction flask, add 15 mL of dichloromethane solution to dissolve it, successively add trichloroacetonitrile (3.22 mL, 32.1 mmol) and DBU (0.10 mL, 0.6 mmol), and stir the reaction at room temperature for 2 h. After the reaction is completed, concentrate it, and purify it by column chromatography (PE:EA = 20:1 - 4:1) to obtain 710 mg of a pale yellow liquid with a yield of 45%. 1 H NMR (400 MHz, CDCl3) δ 8.69 (s, 1H), 6.55 (d, J = 4.0 Hz, 1H), 5.56 (t, J = 9.8 Hz, 1H), 5.22 - 5.09 (m, 2H), 4.37 - 4.05 (m, 3H), 2.10 - 1.99 (m, 12H).

[0210] Step 3: Synthesis of (2R,3R,4S,5R,6S)-2-(acetoxymethyl)-6-(4-bromophenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (Intermediate 32-4)

[0211] Weigh intermediate 32-3 (736 mg, 1.5 mmol), 4-bromophenol (207 mg, 1.2 mmol), molecular sieve (1.00 g) into a 50 mL reaction flask, add 15 mL of dichloromethane solution to dissolve it, slowly dropwise add boron trifluoride diethyl ether complex (0.15 mL, 1.2 mmol) at -30 °C and stir the reaction for 5 h. After the reaction is completed, add 1 M sodium hydroxide solution for extraction, combine the organic phases, dry with anhydrous Na2SO4, and purify it by column chromatography (PE:EA = 15:1 - 2:1) to obtain 223 mg of a white solid with a yield of 37%. 1 H NMR (400 MHz, CDCl3) δ 7.43 - 7.36 (m, 2H), 6.91 - 6.84 (m, 2H), 5.34 - 5.21 (m, 2H), 5.16 (t, J = 9.2 Hz, 1H), 5.03 (d, J = 7.2 Hz, 1H), 4.28 (dd, J = 12.0, 5.4 Hz, 1H), 4.16 (dd, J = 12.4, 2.4 Hz, 1H), 3.85 (ddd, J = 10.0, 5.4, 2.4 Hz, 1H), 2.05 (dd, J = 11.2, 6.4 Hz, 12H).

[0212] Step 4: Synthesis of (2R,3R,4S,5R,6S)-2-(acetoxymethyl)-6-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyl triacetate (Intermediate 32-5)

[0213] Weigh intermediate 32-4 (500 mg, 1.0 mmol), bis(pinacolato)diboron (353 mg, 1.4 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (Pd(dppf)2Cl2) (97 mg, 0.1 mmol), and potassium acetate (292 mg, 3.0 mmol) into a 50 mL reaction flask, add 10 mL of dioxane solution to dissolve, and reflux overnight under argon protection. After the reaction is completed, evaporate the solvent, and purify by column chromatography (DCM:MeOH = 150:1 - 100:1) to obtain 600 mg of a yellow liquid with a yield of 40%. 1 H NMR (400 MHz, DMSO-d6) δ 7.63 (d, J = 8.6 Hz, 2H), 6.99 (d, J = 8.6 Hz, 2H), 5.63 (d, J = 8.0 Hz, 1H), 5.40 (d, J = 9.6 Hz, 1H), 5.13 - 4.95 (m, 2H), 4.31 - 4.11 (m, 2H), 4.08 - 4.00 (m, 1H), 2.06 - 1.90 (m, 12H), 1.28 (s, 12H).

[0214] Step 5: Synthesis of (E)-3-(2-fluorophenyl)-N-(4'-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-[1,1'-biphenyl]-4-yl)acrylamide (Compound 32)

[0215] Weigh the intermediate 32-5 (392 mg, 0.7 mmol), (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide (152 mg, 0.5 mmol), 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium(II) dichloromethane complex (Pd(dppf)2Cl2·CH2Cl2) (78 mg, 0.1 mmol), and potassium phosphate (252 mg, 1.2 mmol) into a 50 mL reaction flask. Add 10 mL of dioxane-distilled water (4:1) solution to dissolve them. After protecting with argon, reflux the reaction for 15 h. After the reaction is completed, evaporate the solvent. The product after column chromatography purification (DCM:MeOH = 150:1 - 80:1) is dissolved in 8 mL of methanol solution, and then sodium methoxide (54 mg, 1.0 mmol) is slowly added. Stir the reaction at room temperature for 1 h. After the reaction is completed, evaporate the solvent and purify by column chromatography (DCM:MeOH = 50:1 - 5:1) to obtain 98 mg of a pale yellow solid with a yield of 99%. 1 H NMR (400 MHz, DMSO-d6) δ 10.39 (s, 1H), 7.82 - 7.76 (m, 2H), 7.72 (td, J = 7.8, 1.6 Hz, 1H), 7.68 - 7.56 (m, 5H), 7.50 - 7.45 (m, 1H), 7.37 - 7.26 (m, 2H), 7.14 - 7.07 (m, 2H), 6.97 (d, J = 15.8 Hz, 1H), 5.33 (d, J = 4.8 Hz, 1H), 5.10 (d, J = 4.4 Hz, 1H), 5.04 (d, J = 5.2 Hz, 1H), 4.90 (d, J = 7.2 Hz, 1H), 4.59 (t, J = 5.8 Hz, 1H), 3.71 (ddd, J = 12.0, 5.6, 2.0 Hz, 1H), 3.52 - 3.44 (m, 1H), 3.30 - 3.24 (m, 2H), 3.22 - 3.15 (m, 2H); HRMS (ESI) m / z calculated for C 27 H 26 FNNaO7 [M + Na] + 518.1586, found 518.1578.

[0216] Example 33: Preparation of (E)-3-(2-fluorophenyl)-N-(5-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)pyridin-2-yl)acrylamide (Compound 33)

[0217]

[0218] Adapt to change the raw material compound (replace the raw material of (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide in step 5 of Example 32 with the raw material of (E)-N-(5-bromopyridin-2-yl)-3-(2-fluorophenyl)acrylamide), and the remaining steps are the same as in Example 32, pale yellow solid, yield 71%. 1 H NMR(400MHz,DMSO-d6)δ10.90(s,1H),8.65(d,J=2.6Hz,1H),8.31(d,J=8.6Hz,1H),8.10(dd,J=8.4,2.4Hz,1H),7.75-7.64(m,4H),7.53-7.43(m,1H),7.37-7.27(m,2H),7.21-7.11(m,2H),5.35(d,J=4.6Hz,1H),5.11(d,J=4.4Hz,1H),5.04(d,J=5.2Hz,1H),4.93(d,J=7.0Hz,1H),4.59(t,J=5.6Hz,1H),3.71(ddd,J=11.8,5.4,2.0Hz,1H),3.48(dt,J=11.8,6.0Hz,1H),3.40-3.34(m,1H),3.28(d,J=4.6Hz,2H),3.21-3.15(m,1H);HRMS(ESI)m / z calculated for C 26 H 26 FN2NaO7[M+Na] + 497.1719,found 497.1710.

[0219] Example 34: Preparation of 2-(2-fluorophenoxy)-N-(4'-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-[1,1'-biphenyl]-4-yl)acetamide (Compound 34)

[0220]

[0221] Adapt to change the raw material compound (replace the raw material of (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide in step 5 of Example 32 with the raw material of N-(4-bromophenyl)-2-(2-fluorophenoxy)acetamide), and the remaining steps are the same as in Example 32, pale yellow liquid, yield 30%. 11H NMR (400 MHz, DMSO-d6) δ 10.24 (s, 1H), 7.69 (d, J = 8.8 Hz, 2H), 7.63 - 7.54 (m, 4H), 7.30 - 7.20 (m, 1H), 7.16 - 7.06 (m, 4H), 7.02 - 6.95 (m, 1H), 5.33 (d, J = 4.4 Hz, 1H), 5.10 (d, J = 4.4 Hz, 1H), 5.03 (d, J = 5.2 Hz, 1H), 4.89 (d, J = 7.2 Hz, 1H), 4.81 (s, 2H), 4.58 (s, 1H), 3.75 - 3.66 (m, 1H), 3.53 - 3.34 (m, 2H), 3.32 - 3.13 (m, 3H); HRMS (ESI) m / z calculated for C 26 H 26 FNNaO8 [M+Na] + 522.1535, found 522.1531.

[0222] Example 35: Preparation of 2-(2-fluorophenoxy)-N-(5-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)pyridin-2-yl)acetamide (Compound 35)

[0223]

[0224] Adapt to change the starting compound (replace the starting material (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide in Step 5 of Example 32 with N-(5-bromopyridin-2-yl)-2-(2-fluorophenoxy)acetamide), and the remaining steps are the same as in Example 32. White solid, yield 79%. 1 1H NMR (400 MHz, DMSO-d6) δ 10.64 (s, 1H), 8.67 - 8.61 (m, 1H), 8.14 - 8.05 (m, 2H), 7.69 - 7.62 (m, 2H), 7.30 - 7.20 (m, 1H), 7.23 - 7.06 (m, 4H), 7.03 - 6.92 (m, 1H), 4.91 (d, J = 1.8 Hz, 2H), 3.70 (dd, J = 11.6, 2.0 Hz, 1H), 3.48 (dd, J = 11.8, 5.6 Hz, 1H), 3.38 - 3.33 (m, 1H), 3.27 (t, J = 8.0 Hz, 2H), 3.17 (t, J = 8.8 Hz, 1H); HRMS (ESI) m / z calculated for C 25 H 25 FN2NaO8 [M+Na]+ 523.1487, found 523.1466.

[0225] Example 36: Preparation of N-(2-fluorophenyl)-2-((4'-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-[1,1'-biphenyl]-4-yl)oxy)acetamide (Compound 36)

[0226]

[0227] Adapt to change the starting compound (replace the starting material (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide in Step 5 of Example 32 with 2-(4-bromophenoxy)-N-(2-fluorophenyl)acetamide), and the remaining steps are the same as in Example 32. Yellow solid, yield 29%. 1 H NMR (400 MHz, DMSO-d6) δ 9.91 (s, 1H), 7.83 (td, J = 7.6, 2.4 Hz, 1H), 7.62 - 7.53 (m, 4H), 7.29 (tt, J = 7.8, 2.6 Hz, 1H), 7.22 - 7.18 (m, 2H), 7.07 (dd, J = 12.2, 8.8 Hz, 4H), 5.32 (d, J = 4.8 Hz, 1H), 5.10 (d, J = 4.4 Hz, 1H), 5.03 (d, J = 5.2 Hz, 1H), 4.88 (d, J = 7.2 Hz, 1H), 4.81 (s, 2H), 4.58 (t, J = 5.6 Hz, 1H), 3.69 (s, 1H), 3.52 - 3.42 (m, 1H), 3.31 - 3.11 (m, 4H); HRMS (ESI) m / z calculated for C 26 H 26 FNNaO8 [M+Na] + 422.1535, found 522.1530.

[0228] Example 37: Preparation of N-(2-fluorophenyl)-2-(5-(4-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)pyridin-2-yl)oxy)acetamide (Compound 37)

[0229]

[0230] Adapt to change the starting compound (replace the starting material (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide in Step 5 of Example 32 with 2-((5-bromopyridin-2-yl)oxy)-N-(2-fluorophenyl)acetamide), and the remaining steps are the same as in Example 32. Yellow solid, yield 40%. 1 H NMR(400MHz,DMSO-d6)δ10.20(s,1H),8.06(t,J=2.4Hz,1H),7.97-7.91(m,1H),7.85(dd,J=9.6,2.8Hz,1H),7.52-7.46(m,2H),7.34-7.22(m,1H),7.19-7.11(m,3H),7.09(d,J=8.8Hz,1H),6.49(d,J=9.4Hz,1H),5.40(s,1H),5.08(dd,J=10.8,4.8Hz,1H),5.04-4.92(m,2H),4.89(t,J=4.4Hz,2H),4.60-4.43(m,1H),3.62(s,2H),3.51-3.33(m,2H),3.31-3.11(m,2H);HRMS(ESI)m / z calculated for C 25 H 25 FN2NaO8[M+Na] + 523.1487,found 523.1488.

[0231] Example 38: Preparation of 3-(2-fluorophenyl)-N-(4'-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-[1,1'-biphenyl]-4-yl)propanamide (Compound 38)

[0232]

[0233] Adapt to change the starting compound (replace the starting material (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide in Step 5 of Example 32 with N-(4-bromophenyl)-3-(2-fluorophenyl)propanamide), and the remaining steps are the same as in Example 32. White solid, yield 45%. 11H NMR (400 MHz, DMSO-d6) δ 10.01 (s, 1H), 7.63 (d, J = 8.4 Hz, 2H), 7.56 (d, J = 8.4 Hz, 4H), 7.34 (td, J = 7.8, 2.0 Hz, 1H), 7.29 - 7.23 (m, 1H), 7.20 - 7.05 (m, 4H), 5.43 - 5.03 (m, 3H), 4.89 (d, J = 7.2 Hz, 1H), 4.60 (s, 1H), 3.70 (d, J = 11.6 Hz, 1H), 3.47 (dd, J = 11.8, 5.6 Hz, 1H), 3.32 - 3.13 (m, 4H), 2.95 (t, J = 7.6 Hz, 2H), 2.65 (t, J = 7.6 Hz, 2H); HRMS (ESI) m / z calculated for C 27 H 28 FNNaO7 [M+Na] + 520.1742, found 520.1758.

[0234] Example 39: Preparation of 3-(2-Fluorophenyl)-N-(5-(4-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)phenyl)pyridin-2-yl)propanamide (Compound 39)

[0235]

[0236] Adapt to change the starting compound (replace the (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide starting material in step 5 of Example 32 with N-(5-bromopyridin-2-yl)-3-(2-fluorophenyl)propanamide starting material), and the remaining steps are the same as in Example 32. Pale yellow solid, yield 28%. 11H NMR (400 MHz, DMSO-d6) δ 10.58 (d, J = 2.2 Hz, 1H), 8.58 (d, J = 2.4 Hz, 1H), 8.14 (d, J = 8.8 Hz, 1H), 8.04 (dd, J = 8.8, 2.6 Hz, 1H), 7.67 - 7.60 (m, 2H), 7.34 (td, J = 7.6, 2.0 Hz, 1H), 7.26 - 7.21 (m, 1H), 7.20 - 7.06 (m, 4H), 5.48 - 4.87 (m, 4H), 4.53 (d, J = 39.2 Hz, 1H), 3.72 - 3.54 (m, 2H), 3.50 - 3.44 (m, 2H), 3.28 - 3.13 (m, 2H), 2.94 (t, J = 7.6 Hz, 2H), 2.73 (t, J = 8.0 Hz, 2H); HRMS (ESI) m / z calculated for C 26 H 27 FN2NaO7 [M+Na] + 521.1695, found 521.1692.

[0237] Example 40: Preparation of 3-(2-fluorophenyl)-N-(5-(4-(2-((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)phenyl)pyridin-2-yl)propanamide (Compound 40)

[0238]

[0239] Adapt to change the raw materials (replace the p-bromophenol raw material in step 3 of Example 32 with 4-(2-hydroxyethyl)phenol raw material, and replace the (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide raw material in step 5 with N-(5-bromopyridin-2-yl)-3-(2-fluorophenyl)propanamide raw material), and the remaining steps are the same as in Example 32, white solid, yield 20%. 11H NMR(400MHz,DMSO-d6)δ10.62(s,1H),8.60(d,J=2.4Hz,1H),8.14(d,J=8.4Hz,1H),8.06(dd,J=8.8,2.6Hz,1H),7.61(d,J=7.8Hz,2H),7.35(dd,J=18.4,8.0Hz,3H),7.30 - 7.20(m,2H),7.14(q,J=8.8,7.2Hz,2H),5.04 - 4.95(m,4H),4.53(t,J=6.0Hz,1H),4.20(d,J=7.6Hz,1H),4.00 - 3.91(m,1H),3.66(dd,J=12.6,6.8Hz,2H),3.07(t,J=6.0Hz,3H),2.93(dt,J=16.4,6.8Hz,5H),2.74(t,J=8.0Hz,2H); HRMS(ESI)m / z calculated forC 28 H 31 FN2NaO7[M+Na] + 549.2008,found 549.2010.

[0240] Example 41: Preparation of (E)-3-(2-fluorophenyl)-N-(4'-(2-((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)-[1,1'-biphenyl]-4-yl)acrylamide (Compound 41)

[0241]

[0242] Adapt to change the starting compound (replace the p-bromophenol starting material in Step 3 of Example 32 with 4-(2-hydroxyethyl)phenol starting material), and the remaining steps are the same as in Example 32. White solid, yield 40%. 11H NMR (400 MHz, DMSO-d6) δ 10.40 (s, 1H), 7.83 - 7.76 (m, 2H), 7.73 (td, J = 7.8, 1.6 Hz, 1H), 7.70 - 7.60 (m, 3H), 7.61 - 7.54 (m, 2H), 7.50 - 7.45 (m, 1H), 7.38 - 7.24 (m, 4H), 6.97 (d, J = 15.8 Hz, 1H), 5.00 (d, J = 5.2 Hz, 2H), 4.92 (dd, J = 14.4, 4.8 Hz, 1H), 4.50 (t, J = 5.6 Hz, 1H), 4.21 (d, J = 7.8 Hz, 1H), 3.98 (dt, J = 9.8, 7.4 Hz, 1H), 3.74 - 3.63 (m, 2H), 3.47 - 3.41 (m, 1H), 3.20 - 2.82 (m, 6H); HRMS (ESI) m / z calculated for C 29 H 30 FNNaO7 [M + Na] + 546.1899, found 546.1890.

[0243] Example 42: Preparation of (E)-3-(2-fluorophenyl)-N-(5-(4-(2-((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)phenyl)pyridin-2-yl)acrylamide (Compound 42)

[0244]

[0245] Adapt to change the starting compounds (replace the p-bromophenol starting material in Step 3 of Example 32 with 4-(2-hydroxyethyl)phenol starting material, and replace the (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide starting material in Step 5 with (E)-N-(5-bromopyridin-2-yl)-3-(2-fluorophenyl)acrylamide starting material), and the remaining steps are the same as in Example 32. White solid, yield 40%. 11H NMR (400 MHz, DMSO-d6) δ 10.92 (s, 1H), 8.67 (d, J = 2.6 Hz, 1H), 8.32 (d, J = 9.6 Hz, 1H), 8.12 (dd, J = 8.8, 2.4 Hz, 1H), 7.76 - 7.59 (m, 4H), 7.51 - 7.46 (m, 1H), 7.42 - 7.37 (m, 2H), 7.35 - 7.28 (m, 2H), 7.17 (d, J = 15.6 Hz, 1H), 5.03 - 4.87 (m, 3H), 4.49 (t, J = 6.0 Hz, 1H), 4.21 (d, J = 8.0 Hz, 1H), 3.99 (dt, J = 14.8, 7.4 Hz, 1H), 3.75 - 3.63 (m, 2H), 3.43 (dt, J = 11.6, 6.0 Hz, 1H), 3.20 - 2.83 (m, 6H); HRMS (ESI) m / z calculated for C 28 H 29 FN2NaO7 [M+Na] + 547.1851, found 547.1855.

[0246] Example 43: Preparation of 2-(2-fluorophenoxy)-N-(5-(4-(2-(((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)phenyl)pyridin-2-yl)acetamide (Compound 43)

[0247]

[0248] Adapt to change the starting materials (replace the starting material of p-bromophenol in step 3 of Example 32 with 4-(2-hydroxyethyl)phenol, and replace the starting material of (E)-N-(4-bromophenyl)-3-(2-fluorophenyl)acrylamide in step 5 with N-(5-bromopyridin-2-yl)-2-(2-fluorophenoxy)acetamide), and the remaining steps are the same as in Example 32. White solid, yield 21%. 1HNMR(400MHz, DMSO-d6) δ 10.67 (s, 1H), 8.66 (t, J = 1.6 Hz, 1H), 8.11 (d, J = 2.2 Hz, 2H), 7.66 - 7.60 (m, 2H), 7.30 - 7.20 (m, 1H), 7.15 - 7.07 (m, 2H), 7.02 - 6.92 (m, 1H), 5.00 (d, J = 5.0 Hz, 1H), 4.96 - 4.87 (m, 4H), 4.49 (t, J = 6.0 Hz, 1H), 4.20 (d, J = 7.8 Hz, 1H), 3.98 (dt, J = 14.8, 7.4 Hz, 1H), 3.74 - 3.63 (m, 2H), 3.48 - 3.37 (m, 1H), 3.16 - 2.93 (m, 4H), 2.89 (d, J = 8.2 Hz, 2H); HRMS(ESI) m / z calculated for C 27 H 29 FN2NaO8 [M+Na] + 551.1800, found 551.1815.

[0249] Example 44: Evaluation of the in vitro HIF-1α HRE transcriptional inhibitory activity of the salidroside derivatives of the present invention

[0250] The dual-luciferase reporter assay was used to detect the transcriptional inhibitory activity of the salidroside derivatives of the present invention against HIF-1α HRE.

[0251] Experimental procedure: HEK293T cells were cultured to the logarithmic phase and seeded in 96-well plates at a density of 6×10 3 cells per well. The plates were placed in a 37 °C cell culture incubator containing 5% CO2 and incubated for 24 h until the cells adhered. Lipofectamine 2000 transfection reagent was used to transfect the cells with plasmids expressing hypoxia response element (HRE)-dependent firefly luciferase reporter gene and CMV-renilla luciferase reporter gene. Under normoxic and hypoxic conditions (1% O2, 94% N2, and 5% CO2), the cells were treated with 20 μM of the target compound for 12 h. Luciferase detection was performed using a luciferase detection system (Promega, Madison, WI) according to the manufacturer's instructions. The fluorescence intensity of firefly luciferase was calculated using renilla fluorescence as an internal reference, and the experiment was repeated independently three times.

[0252] The experimental results of the transcriptional inhibitory activity of the salidroside derivatives of the present invention against HIF-1α HRE are shown in Table 1 as follows:

[0253] Table 1 Experimental results of the evaluation of the in vitro anti-tumor cell proliferation activity of the target compound

[0254]

[0255] a The transcriptional inhibitory activity of HIF-1α HRE in HEK293T cells was detected by the Dual-Luciferase method at a dosing concentration of 20 μM of the target compound, with three independent parallel experiments.

[0256] b The test target compound has no transcriptional inhibitory activity on HIF-1α HRE.

[0257] As can be seen from Table 1, most of the salidroside derivatives described in the present invention have moderate to strong inhibitory activity against HIF-1α. At a dosing concentration of 20 μM, the transcriptional inhibition rates of compounds 1-5, compound 7, compound 22, compound 23, compounds 25-27, compound 33, compound 41, and compound 42 on HIF-1α HRE are greater than 50%; among them, the transcriptional inhibition rates of compounds 4, compound 22, compound 23, compound 25, compound 26, and compound 41 on HIF-1α HRE are greater than 70%; in particular, the transcriptional inhibition rates of compound 23 and compound 41 on HIF-1α HRE are greater than 90%, laying an active foundation for the subsequent in-depth study of the mechanism of action and the indications of acute and chronic altitude diseases.

[0258] Example 45: Evaluation of the in vitro cytotoxic activity of the salidroside derivatives described in the present invention

[0259] The anti-proliferative activity of the salidroside derivatives described in the present invention against astrocytic C8-D1A cells was detected using the CCK-8 method.

[0260] Experimental procedure: Culture C8-D1A cells to the logarithmic phase, seed 7×10 3 cells per well in a 96-well plate, and place it in a 37 °C cell incubator containing 5% CO2 for 24 h until the cells adhere. Then add a medium containing 20 μM of the target compound and co-culture for 72 h, with three parallel replicates. After the culture is completed, add 10 μL of CCK-8 solution and incubate for another 3 h. Discard the suspension, measure the absorbance value of the cell lysate at a wavelength of 450 nm using a multifunctional microplate reader, and calculate the inhibition rate using GraphPad Prism software.

[0261] As Figure 1 shown, most of the salidroside derivatives described in the present invention have no significant cytotoxicity to astrocytic C8-D1A cells at a concentration of 20 μM. In particular, the cell viabilities of compounds 10, 11, 30, 31, 37, and 38 on C8-D1A are comparable to those of the control group, showing good in vitro safety.

[0262] The synthesis method of this kind of compound is simple, the raw materials are cheap and easily available, and it has significant HIF-1α HRE transcriptional inhibitory activity. It is expected to become a new drug for the prevention and / or treatment of acute and chronic altitude diseases such as acute high altitude cerebral edema, acute high altitude pulmonary edema, high altitude polycythemia, high altitude pulmonary hypertension, and altitude decline disease.

[0263] The above embodiments are only exemplary descriptions of the present invention, and those skilled in the art can modify or optimize them according to actual needs. It should be particularly pointed out that as long as they do not deviate from the core idea or basic principle of the present invention, such modifications or optimizations should be regarded as belonging to the protection scope of the present invention.

Claims

1. A salidroside derivative having hypoxia-inducible factor-1α (HIF-1α) protein inhibitory activity, or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, polymorph, metabolite or isotopically labeled compound thereof, wherein the structure of the derivative is shown in formula (I): In formula (I): Ring A is absent, or a substituted / unsubstituted benzene ring or pyridine ring; X is selected from oxyacetyl ester group, acryloyl ester group, oxyacetylamino group, 2-butenamide group, acetamide group, propionamide group, acrylamide group, 1-propenamido group, carbamoyl group, ethylcarbamoyl group, hydrazide group, propenyl group, mercaptoacetamide group, oxyacetamide group, oxyacrylamide group, 3-(1-propenyl)-3,4,5-triazolyl group; preferably acryloyl ester group, acrylamide group, acetamide group, oxyacetamide group; R 1 is substituted by hydrogen or glucosyl; R 2 is substituted by hydrogen or 2-fluoro, 4-methoxy-3-hydroxy, 4-hydroxy, 3-methoxy-4-hydroxy; n is 0 or 1, indicating absent or containing one saturated ethyl group.

2. The salidroside derivative according to claim 1, wherein: At least one selected from the following segments: At least one selected from the following segments: R 1 At least one selected from the following fragments: H、 At least one selected from the following segments:

3. The salidroside derivative according to claim 1 or 2, characterized in that, The derivative is a compound represented by formula (II), (III), (IV) or (V): Wherein, X and R 2 The substituents are respectively as described in Claim 1 or 2; in formula (IV) or (V), n is 0 or 1, indicating the absence or presence of one saturated ethyl group, and Y is a C atom or an N atom.

4. The salidroside derivative according to claim 1, wherein The salidroside derivative represented by formula (I) is one of the following compounds:

5. The synthetic method of the salidroside derivative according to any one of claims 1-4, characterized in that, Including one of the following synthetic routes: Synthetic route 1: Substituted phenol (II-1) reacts with methyl bromoacetate through nucleophilic substitution reaction to obtain intermediate II-2, and under strong alkaline conditions, intermediate II-2 undergoes ester hydrolysis reaction to obtain carboxylic acid intermediate II-3; Carboxylic acid intermediate II-3 reacts with β-D-glucose through Mitsunobu reaction under the action of a coupling reagent to prepare the compound of formula (II); Synthetic route 2: 4-Nitrophenol (III-1) undergoes a nucleophilic substitution reaction with (3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate to obtain intermediate III-2; intermediate III-2 is subjected to a nitro reduction reaction under the action of a reducing agent to obtain nitro reduction product intermediate III-3; intermediate III-3 further reacts with an R 2 substituted phenyl acid through an amide condensation reaction to obtain intermediate III-4; p-Acetoxybenzoic acid (III-5) reacts with R 2 substituted phenylamino side chain through amide condensation reaction to obtain III-6; Intermediate III-6 is deacetylated under strong alkaline conditions to obtain III-7; Intermediate III-7 further undergoes nucleophilic substitution reaction with (3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate to obtain intermediate III-8; R 2 The substituted aniline (III-9) undergoes a nucleophilic substitution reaction with bromoacetyl bromide to obtain the intermediate III-10; the intermediate III-10 undergoes a nucleophilic substitution reaction with p-methoxythiophenol or 4-methoxyphenol under strongly basic conditions to obtain the intermediate III-11; (E)-3-(4-methoxyphenoxy)acrylic acid (III-12) reacts with R 2 the substituted aniline through an amide condensation reaction to obtain the intermediate III-13; R 2 the substituted (E)-(3-azidoprop-1-en-1-yl)benzene (intermediate III-14) undergoes a Click reaction with 4-methoxyphenylacetylene to obtain the intermediate III-15; the intermediates III-11, III-13, and III-15 are prepared to obtain the intermediate III-16 through a demethylation reaction; the intermediate III-16 further undergoes a nucleophilic substitution reaction with (3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate to obtain the intermediate III-17; Intermediates III-4, III-8, III-17 remove the acetyl group under strong alkaline conditions to obtain the compound of formula (III); Synthetic route 3: 4-bromophenol or 4-bromophenylethanol (IV-1) reacts with bis(pinacolato)diboron through Miyaura borylation reaction under the action of a catalyst to obtain borate intermediate IV-2; The arylamine or aryl phenolic acid (IV-3) substituted by Y atom reacts with R 2 The substituted aryl side chain undergoes an amide condensation reaction to obtain the intermediate IV-5, or the aryl phenolic acid (IV-4) substituted by Y atom reacts with R 2 The substituted aryl side chain undergoes a nucleophilic substitution reaction under strong basic conditions to obtain the intermediate IV-5; Intermediate IV-5 and intermediate IV-2 undergo Suzuki-Miyaura cross-coupling reaction to obtain the compound of formula (IV); Synthetic route 4: (3R,4S,5S,6R)-6-(acetoxymethyl)tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate (V-1) removes the 2-position acetyl group under alkaline conditions to obtain intermediate V-2; Intermediate V-2 reacts with trichloroacetonitrile through Pinner reaction under alkaline conditions to obtain intermediate V-3; Intermediate V-3 reacts with 4-bromophenol or 4-bromophenylethanol through nucleophilic substitution reaction to obtain intermediate V-4; Intermediate V-4 reacts with bis(pinacolato)diboron through Miyaura borylation reaction under the action of a catalyst to obtain borate intermediate V-5; Intermediate V-5 and intermediate IV-5 undergo Suzuki-Miyaura cross-coupling reaction to obtain the compound of formula (V).

6. A pharmaceutical composition, characterized in that, Comprising the salidroside derivative according to any one of claims 1-4 or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, polymorph, metabolite, isotopically labeled compound, enantiomer, diastereomer or tautomer thereof, and at least one pharmaceutically acceptable carrier.

7. A pharmaceutical preparation comprising the salidroside derivative according to any one of claims 1-4 or the pharmaceutical composition according to claim 6.

8. Use of the salidroside derivative according to any one of claims 1-4, the pharmaceutical composition according to claim 6, or the pharmaceutical preparation according to claim 7 in the preparation of an HIF-1α protein inhibitor.

9. Use of the salidroside derivative according to any one of claims 1-4, the pharmaceutical composition according to claim 6, or the pharmaceutical preparation according to claim 7 in the preparation of an anti-hypoxia or anti-inflammatory drug.

10. Use of the salidroside derivative according to any one of claims 1 to 4, the pharmaceutical composition according to claim 6, or the pharmaceutical preparation according to claim 7 in the preparation of a drug for preventing or treating altitude sickness associated with overexpression of HIF-1α, characterized in that, The altitude diseases include: Acute altitude diseases: acute altitude cerebral edema, acute altitude pulmonary edema, mixed acute altitude disease; Chronic altitude diseases: altitude polycythemia, high altitude pulmonary hypertension, abnormal altitude blood pressure, altitude heart disease, altitude decline syndrome, mixed chronic altitude disease.

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