Double-activated hydroxyl light cage group modified galactoside anti-tumor prodrug and application thereof

Abstract

The invention discloses a double-activated hydroxyl light cage group modified galactoside anti-tumor prodrug and application thereof, and belongs to the field of organic chemistry and medicinal chemistry. The invention provides a prodrug compound with a structure shown as the following: # imgabs0 # R3, R4, R5 and R6, any one of which is a light cage group capable of breaking an ether bond and releasing galactose hydroxyl after illumination, and the other three of which are H. The prodrug compound takes beta-galactosidase as a target spot, and a modification group which can be activated and dissociated by illumination is introduced into galactose hydroxyl, so that the space-time controllability of a galactoside single prodrug release drug is further improved, and the toxic and side effects are reduced.

Description

Technical Field

[0001] The invention belongs to the field of organic chemistry and medicinal chemistry, and specifically relates to a dual-activated hydroxyl photocage group-modified galactoside anti-tumor prodrug and application thereof. Background Art

[0002] β-galactoside prodrugs are oxygen glycoside compounds obtained by coupling the terminal of β-D-galactose with the active essential group of the drug molecule. The active drug can be released after the β-galactosidase hydrolyzes the β-galactosidase bond. Compared with normal cells, the activity of β-galactosidase in tumor cells such as ovarian cancer, colon cancer, and breast cancer is increased. Therefore, β-galactoside prodrugs are expected to release drugs in tumor cells in a controllable manner and selectively kill tumor cells. In the past three decades, dozens of β-galactoside prodrugs with different structures for anti-tumor use have been reported.

[0003] For example, as early as 1994, there were reports of galactoside prodrugs of 5-fluorouracil. Since 2002, the Tietze team at the University of Göttingen in Germany has developed a series of galactoside prodrugs of duocarcin derivatives, including. Since 2011, the Papot team at the University of Poitiers in France has successively reported a series of galactoside anti-tumor prodrugs with functional ligands (such as folic acid, monoclonal antibodies, etc.), which can release drugs such as doxorubicin and monomethyl auristatin E (MMAE). The corresponding galactoside prodrugs of 8-hydroxyquinoline derivatives (HQ), Compretin (CA4), gemcitabine, and aminonafide have also been reported. Among domestic scholars, Wang Peng's team reported the galactoside prodrug of geldanamycin in 2005; Huang Gangliang's team at Chongqing Normal University reported the galactoside prodrug of epothilone in 2018; and Hou Jingli's team at Tianjin Medical University reported a galactoside prodrug that can release hydrogen sulfide in 2024. These reported β-galactoside prodrugs are all prodrugs activated by a single stimulus condition (β-galactosidase), referred to as "single prodrugs".

[0004] However, in addition to inhibiting certain tumor cells, these β-galactosidase single prodrugs also produce certain toxicity in non-tumor cells with high expression of β-galactosidase (such as M2 polarized macrophages, high glucose-induced macrophages, neurons, osteoclasts, etc.) and normal tissues (such as liver, small intestine, placenta, etc.). The uncontrollability of their spatiotemporal release makes them have the risk of being activated in non-tumor sites to release drugs and produce toxic side effects. Therefore, it is urgent to provide a prodrug with better spatiotemporal controllability of drug release and low toxic side effects. Summary of the invention

[0005]

Technical issues

[0006] In order to further improve the spatiotemporal controllability of drug release from galactoside single prodrugs and reduce toxic side effects, the present invention provides a β-galactoside prodrug activated by dual stimulation conditions, referred to as a "double prodrug". The double prodrug needs to respond to light and β-galactosidase in sequence to release the drug, and the controllability of drug release is further improved through the logic of "AND gate".

[0007]

Technical solution

[0008] The present invention firstly introduces photocage groups on the 2, 3, 4, and 6 hydroxyl groups of galactoside through orthogonal protection and regioselectivity strategies, and constructs a new dual-activated hydroxyl photocage group-modified galactoside anti-tumor prodrug (such as Figure 1 ). The present invention constructs a total of four new general synthetic routes for galactoside double prodrugs. The synthetic route is mainly divided into three steps: 1) using the hydroxyl orthogonal protection strategy to selectively block part of the hydroxyl groups of galactose, and regioselectively introducing photocage groups on the 2, 3, 4, and 6 hydroxyl groups; 2) introducing a linker arm at the end of galactose through a glycosylation reaction and further coupling a cytotoxic molecule; 3) removing the blocking group of the hydroxyl group on galactose to obtain the final double prodrug. The present invention also compares the drug release controllability and anti-tumor activity of double prodrug positional isomers modified with hydroxyl groups at different positions.

[0009] The galactoside double prodrug in the present invention does not respond to β-galactosidase by itself, and needs to be converted into a single prodrug after first removing the photocage group through light irradiation, and further responds to β-galactosidase to release the drug. This type of new structure of double prodrug has the following advantages: (1) It improves the controllability of drug release and reduces the risk of toxic side effects; (2) Theoretically, all β-galactoside single prodrugs can be constructed into double prodrugs by this method, which has universal applicability; (3) Photocage groups with different optical properties can be introduced on the galactose hydroxyl group, which has high flexibility; (4) The galactoside single prodrug has four exposed hydroxyl groups at positions 2, 3, 4, and 6, which can be introduced with photocage groups, which provides the possibility of constructing galactoside double prodrugs with diverse structures, fully studying the structure-activity relationship, and finding the optimal double prodrug.

[0010] The specific contents of the plan are as follows:

[0011] The first object of the present invention is to provide a β-galactoside anti-tumor prodrug modified with a hydroxyl photocage group having the structure shown below, or a pharmaceutically acceptable salt thereof:

[0012]

[0013] in,

[0014] Z is a vacant bond or an ester group (-CO-O-);

[0015] R 1Derived from cytotoxic drugs containing hydroxyl or amino groups;

[0016] R 2 Selected from hydrogen atoms, C 1-6 Alkyl, halogen, nitro, amino, carboxyl, C 1-6 Alkoxy, acylamino R a -CONH-, alkoxyacyl R b OCO-, where R a , R b Independently selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 3-6 Cycloalkyl, halogenated C 3-6 Cycloalkyl, alkenyl, halogenated alkenyl, aromatic group;

[0017] R 3 , R 4 , R 5 , R 6 Any one of them is a photocage group that can break the ether bond and release the galactose hydroxyl group after light exposure, and the other three are H;

[0018] The photocage group is selected from R c Selected from hydrogen atoms or C 1-6 Alkyl; R d , R e independently selected from hydrogen, halogen, nitro, amino, C 1-6 alkylamino, substituted or unsubstituted five- to six-membered saturated heterocyclic group, substituted or unsubstituted phenyl, substituted or unsubstituted C 1-6 The substituted groups on the alkoxy, saturated heterocyclic group or phenyl group are selected from halogen, nitro, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino; C 1-6 The substituted groups on the alkoxy group are selected from halogen, nitro, C 1-6 Alkyl, halogenated C 1-6 alkyl.

[0019] In one embodiment of the present invention, the prodrug can release the cytotoxic drug in a sequential manner in response to light and β-galactosidase.

[0020] In one embodiment of the present invention, the prodrug has a structure shown in formula (I)-(VIII):

[0021]

[0022] In one embodiment of the present invention, R 1It is derived from a cytotoxic drug containing a hydroxyl group, with the hydroxyl oxygen atom as the connection site. The cytotoxic drug is selected from Compretin (CA4), 8-hydroxyquinoline (HQ), 5-chloro-8-hydroxyquinoline, camptothecin, 7-ethyl-10-hydroxycamptothecin, resorufin, iodoresorufin, duocarmycin MA, duocarmycin MB, duocarmycin GA, duocarmycin TM, chlorambucil, and 5-fluorouridine.

[0023] In one embodiment of the present invention, R in formula (V)-(VIII) 1 It is derived from a cytotoxic drug containing an amino group, with the amino nitrogen atom as the connection site, and the cytotoxic drug is selected from curcumin analog EF24, aminafide, gemcitabine, cytarabine, monomethyl auristatin E (MMAE), methylene blue, cyclophosphamide, linifanib, and doxorubicin.

[0024] Furthermore, C 1-6 The alkylamino group may specifically be a diethylamino group.

[0025] Furthermore, the five- to six-membered saturated heterocyclic group includes at least one heteroatom, and the heteroatom is N, O, or S. The five- to six-membered saturated heterocyclic group specifically includes: morpholinyl, piperidinyl, and piperazinyl.

[0026] Furthermore, C 1-6 Alkoxy groups include: C 1-6 Straight chain alkoxy, C 1-6 of branched alkoxy.

[0027] Preferably, Z is an empty bond, R 1 is the residue of a cytotoxic compound having a hydroxyl group, and the corresponding structure is as follows:

[0028]

[0029] Furthermore, the photocage group is preferably Where R c is selected from a hydrogen atom or a methyl group; R d , R e independently selected from hydrogen, halogen, nitro, amino, hydroxyl, diethylamino, morpholinyl, piperidinyl, substituted or unsubstituted piperazinyl, substituted or unsubstituted phenyl, substituted or unsubstituted C 1-6 Straight chain alkoxy, substituted or unsubstituted C 1-6 of branched alkoxy.

[0030] Furthermore, the prodrug is selected from the following structures:

[0031]

[0032] Further optimization of R c is a hydrogen atom, R dand R e For methoxy:

[0033]

[0034] Preferred R 2 is nitro, more preferably R 2 Replacement position:

[0035]

[0036] Further optimization of R 1 From Compruetin (CA4), the corresponding structural formula is shown below:

[0037]

[0038] In one embodiment of the present invention, the pharmaceutically acceptable salt comprises an inorganic salt or an organic salt; the inorganic salt is selected from sodium salt, calcium salt, potassium salt, magnesium salt, silver salt, lithium salt; the organic salt is selected from meglumine salt, tromethamine salt, diethylamine salt, lysine salt, choline salt, arginine salt, tert-butylamine salt, N,N-dibenzylethylenediamine salt.

[0039] The present invention also provides solvent compounds, enantiomers, diastereomers, tautomers or mixtures in any proportions of the above-mentioned dual-activated galactoside prodrug compounds; specifically including racemic mixtures.

[0040] The present invention also provides the use of the above-mentioned dual-activated galactoside prodrug compound or a pharmaceutically acceptable salt thereof, or a solvent compound, enantiomer, diastereomer, tautomer or a mixture thereof in any proportion of the above-mentioned dual-activated galactoside prodrug compound in the preparation of anti-tumor drugs.

[0041] The present invention also provides a composition or preparation product, which contains the above-mentioned dual-activated galactoside prodrug compound or a pharmaceutically acceptable salt thereof, or a solvent compound, enantiomer, diastereomer, tautomer or a mixture thereof in any proportion of the above-mentioned dual-activated galactoside prodrug compound.

[0042] The composition or preparation is:

[0043] (1) A composition or preparation for preparing a composition or preparation capable of controllably eliminating tumor cells with high expression of β-galactosidase;

[0044] (2) Used for preparing a composition or preparation for improving or treating tumors.

[0045] The composition is a pharmaceutical combination, a dietary supplement, a food composition or a health product composition.

[0046] The present invention also provides an anti-tumor drug, which contains a pharmaceutically acceptable salt, solvent compound, enantiomer, diastereomer, tautomer or a mixture of any proportion of the above-mentioned galactose prodrug compound or the above-mentioned dual-activated galactose prodrug compound, and a pharmaceutically acceptable carrier, excipient or diluent.

[0047] The pharmaceutically acceptable carrier is selected from microcapsules, microspheres, nanoparticles and liposomes.

[0048] The preparation of the drug includes a variety of clinical drug dosage forms, which can be specifically selected from injection, lyophilized powder for injection, suspension, implant, embolic agent, capsule, tablet, pill and oral solution.

[0049] The present invention aims to construct an unreported dual-stimulation condition-activated β-galactoside prodrug. The dual prodrug needs to respond to light and β-galactosidase in sequence to release the drug, and the controllability of drug release is improved through the logic structure of "AND gate". The present invention modifies the 2, 3, 4, and 6 hydroxyl groups of galactose with a photocage group to obtain Figure 1 The double prodrug shown. Since β-galactosidase has a high substrate specificity, the introduction of the photocage group makes the double prodrug unable to be hydrolyzed by β-galactosidase, reducing the risk of releasing the parent drug in non-tumor sites. The double prodrug is converted into a single prodrug after removing the photocage group by light, and further responds to β-galactosidase to release the drug. Activity tests show that the double prodrug prepared by the present invention has strong controllability.

[0050] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0051] (1) The anti-tumor prodrug provided by the present invention is a new type of compound that has never been reported.

[0052] (2) The modification of galactose hydroxyl group in this type of double prodrug compounds improves the controllability of drug release.

[0053] (3) HPLC analysis showed that the modification of the galactose hydroxyl group in compounds A, B, C, and D made the double prodrug compounds non-responsive to β-galactosidase, thus avoiding the problem of insufficient distribution specificity when using β-galactosidase as the response group.

[0054] (4) At the cellular level, the MTT colorimetric method was used to verify that the present invention improved the controllability of drug release. In galactose-overexpressing OVCAR3 and SKOV3 cells, the toxicity of the double prodrugs A, B, C, and D was significantly reduced compared with the single prodrug compounds of the corresponding structures. In addition, the toxicity of the double prodrugs after light activation was improved compared with that before activation.

[0055] (5) At the cellular level, compound A with better controllability was selected as the representative, and the enhanced controllability of the double prodrug was further verified at the cell cycle, cell apoptosis and cell migration experimental levels. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 This is the mechanism by which the galactoside double prodrug of the present invention releases the cytotoxic drug through double activation.

[0057] Figure 2 is the H NMR spectrum of compound A.

[0058] Figure 3 This is the C NMR spectrum of compound A.

[0059] Figure 4 This is the H NMR spectrum of compound B.

[0060] Figure 5 This is the C NMR spectrum of compound B.

[0061] Figure 6 This is the H NMR spectrum of compound C.

[0062] Figure 7 This is the C NMR spectrum of compound C.

[0063] Figure 8 This is the H NMR spectrum of compound D.

[0064] Fig. 9 This is the C NMR spectrum of compound D.

[0065] Fig.10 The HPLC test results show that compounds A, B, C and D do not respond to β-galactosidase.

[0066] Fig.11 These are the results of cytotoxicity tests of compounds A, B, C and D on OVCAR3 cells.

[0067] Fig.12 These are the results of cytotoxicity tests of compounds A, B, C and D on SKOV3 cells.

[0068] Fig.13 This is the cell cycle inhibition result of compound A and the corresponding single prodrug on OVCAR3 cells.

[0069] Fig.14 This is the result of compound A and the corresponding single prodrug promoting apoptosis of OVCAR3 cells.

[0070] Fig.15 This is the result of the scratch healing experiment of compound A and the corresponding single prodrug on OVCAR3 cells. DETAILED DESCRIPTION

[0071] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0072] In the following examples, compound i can be prepared by referring to the literature (Collins, SL; Saha, J.; Bouchez, LC; Hammond, EM; Conway, SJ Hypoxia-Activated, Small-Molecule-Induced Gene Expression. ACS Chem Biol, 2018, 13 (12), 3354-3360.).

[0073] Compound viii can be prepared by referring to the literature (D'Angelo, KA; Taylor, MS Borinic Acid Catalyzed Stereo-and Regioselective Couplings of Glycosyl Methanesulfonates. J Am Chem Soc, 2016, 138(34), 11058-11066.).

[0074] Compound xv can be prepared by referring to the literature (Liang, C.; Lee, DW; Newton, MG; Chu, CK Synthesis of L-Dioxolane Nucleosides and Related Chemistry. J Org Chem, 1995, 60(6), 1546-1553).

[0075] Compound xxii can be prepared by referring to the literature (Chauvin, T.; Durand, P.; Bernier, M.; Meudal, H.; Doan, B.-T.; Noury, F.; Badet, B.; Beloeil, J.-C.; Tóth, é. Detection of Enzymatic Activity by PARACEST MRI: A General Approach to Target a Large Variety of Enzymes. Angew Chem Int Edit, 2008, 47 (23), 4370-4372.).

[0076] Compound xxvi can be found in the literature (Tang, S.; Wan, Z.; Gao, Y.; Zheng, J.-S.; Wang, J.; Si, Y.-Y.; Chen, X.; Qi, H.; Liu, L.; Liu, W. Sci,2016,7(3),1891-1895.) Self-made.

[0077] Other reagents used were commercially purchased.

[0078] Example 1: Synthesis of Prodrug A

[0079] (1) Synthesis of compound ii:

[0080]

[0081] Specific operations:

[0082] Under inert gas protection, compound i (1.12 g, 2.17 mmol, 1.0 equiv.) was dissolved in dichloromethane (20 mL), and silver oxide (571 mg, 8.39 mmol, 2.0 equiv.), tetrabutylammonium iodide (402 mg, 1.09 mmol, 0.5 equiv.), and Molecular sieves. After stirring for 10 minutes, 4,5-dimethoxy-2-nitrobenzyl bromide (898 mg, 3.25 mmol, 1.5 equiv.) dissolved in dichloromethane (5 mL) was added, and the reaction solution was stirred at room temperature for 12 hours. After the reaction was completed, the reaction solution was diluted with dichloromethane and the silver oxide was filtered using diatomaceous earth. The organic phase was washed twice in sequence with water, saturated ammonium chloride and saturated brine, respectively. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated by reduced pressure distillation using a rotary evaporator to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 20:1, v / v) to obtain a yellow oil compound ii (649 mg, 0.911 mmol, yield 42%).

[0083] 1 H NMR (400 MHz, CDCl 3 )δ7.72-7.67(m,4H,Ar),7.65(s,1H,Ar(DMNB)),7.50(s,1H,Ar(DMNB)),7.47-7.34(m,6H,Ar),5.29-5.14(m,2H,ArCH2 ),4.52(d,J=9.9Hz,1H,1-H),4.28(dd,J=5.5,2.1Hz,1H,4-H),4.22(dd,J=6.8,5.4Hz,1H,3-H),4.03(s,3H,ArOCH 3 ),3.96-3.89(m,5H,6-H,ArOCH 3 ),3.84(ddd,J=7.1,5.8,2.1Hz,1H,5-H),3.47(dd,J=9.9,6.7Hz,1H,2-H),3.27(hept,J=6.7Hz,1H,SCH),1.39(s,3H,CH 3 (isopropylidene)),1.34-1.27(m,9H,CH 3 (isopropylidene),CH 3 (isopropylthio)×2),1.05(s,9H,C(CH 3 ) 3 (TBDPS)).

[0084] 13 C NMR (151 MHz, CDCl 3 )δ153.7,147.6,139.6,135.7,135.6,133.5,133.4,130.7,129.8,127.8,127.7,110.5,110.1,107. 7,82.6,80.1,79.4,73.7,69.4,63.0,56.6,56.4,34.3,28.0,26.81,26.79,26.4,23.9,23.7,19.2.

[0085] ESI-MS: found 734.20[M+Na] + .

[0086] (2) Synthesis of Compound iv:

[0087]

[0088] Specific operations:

[0089] (a) Under argon protection, compound ii (300 mg, 0.421 mmol, 1.0 equiv.) was dissolved in dry dichloromethane (3 mL). The mixture was cooled to 0° C. in an ice bath and stirred for 10 minutes. Then, Molecular sieves and iodine monobromide (87 mg, 0.421 mmol, 1.0 equiv.). After the reaction is completed, the molecular sieves are removed by filtration, and the organic phase is collected and washed twice in sequence with 10% sodium thiosulfate, saturated sodium bicarbonate and saturated brine. The organic phase is then dried with anhydrous sodium sulfate, filtered, and then concentrated with a rotary evaporator under reduced pressure to obtain a crude bromide, which is directly used in the next step.

[0090] (b) The crude bromide was dissolved in dry acetonitrile (4 mL), and compound iii (85 mg, 0.506 mmol, 1.2 equiv.), silver oxide (195 mg, 0.842 mmol, 2.0 equiv.), tetrabutylammonium iodide (78 mg, 0.211 mmol, 0.5 equiv.) and Molecular sieve. After reacting at room temperature for 12 hours, the silver oxide and molecular sieve were filtered using diatomaceous earth, and the organic phase was washed twice with sodium carbonate, water, saturated sodium bicarbonate and saturated brine in sequence. The organic phase was collected and dehydrated with anhydrous sodium sulfate, and the solvent was concentrated after filtering out the sodium sulfate solid to obtain a crude mixture. The crude product was separated and purified using a silica gel column (petroleum ether: ethyl acetate, 7:3, v / v) to obtain a light yellow oily compound iv (142 mg, 0.180 mmol, yield 42%).

[0091] 1 H NMR (400 MHz, CDCl 3 )δ9.89(s,1H,ArCHO),8.31(t,J=1.6Hz,1H,Ar),7.79(dt,J=8.7,1.6Hz,1H,Ar),7.7 1-7.60(m,5H,Ar×4,Ar(DMNB)),7.48-7.28(m,8H,Ar×7,Ar(DMNB)),5.24(s,2H,ArCH 2 ),5.13(d,J=7.8Hz,1H,1-H),4.33(t,J=6.1Hz,1H,3-H),4.24(dd,J=5.6,1.8Hz,1H,4-H),4.07(t,J=6.1Hz,1H,5-H),4.03-3.90(m,8H,6-H,ArOCH 3 ×2),3.77(t,J=7.2Hz,1H,2-H),1.40(s,3H,CH 3 (isopropylidene)),1.31(s,3H,CH 3 (isopropylidene)),1.07(s,9H,C(CH 3 ) 3 (TBDPS)).

[0092] 13 C NMR (101 MHz, CDCl 3 )δ188.7,154.1,153.4,147.9,140.7,140.2,135.62,135.56,134.4,133.2,132.9,130.6,130.0,129.9,129.3,127.84,1 27.80,126.9,118.0,111.3,110.8,108.1,100.5,79.7,78.5,75.0,73.1,70.6,63.0,56.5,56.4,27.8,26.8,26.3,19.3.

[0093] ESI-MS: found 825.19[M+Na] + .

[0094] (3) Synthesis of Compound V:

[0095]

[0096] Specific operations:

[0097] Under the protection of inert gas, compound iv (130 mg, 0.162 mmol, 1.0 equiv.) was dissolved in a mixed solution of dichloromethane and isopropanol (2 mL), the mixed solution was cooled to 0 ° C in an ice bath, sodium borohydride powder (12 mg, 0.324 mmol, 2.0 equiv.) was added, and the reaction solution was slowly heated from 0 ° C to room temperature and continued to react for 2 hours. After TLC confirmed that the crude raw material reaction was complete. Then the reaction solution was diluted with dichloromethane, and washed twice in sequence with water, saturated ammonium chloride, saturated sodium bicarbonate, and saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, and the anhydrous sodium sulfate was filtered off and the organic phase was concentrated by vacuum distillation using a rotary evaporator. After concentration, the crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 1: 1, v / v) to obtain a light yellow oily compound v (123 mg, 0.153 mmol, yield 94%).

[0098] 1 H NMR (600 MHz, CDCl 3 )δ7.80(s,J=2.0Hz,1H,Ar),7.72-7.61(m,5H,Ar×4,Ar(DMNB)),7.48-7.24(m,9H,Ar×8,Ar(DMNB)),5.25(s,2H,ArCH 2 (DMNB)),5.01(d,J=7.9Hz,1H,1-H),4.68-4.61(m,2H,ArCH 2),4.30(dd,J=6.7,5.5Hz,1H,3-H),4.23(dd,J=5.6,1.9Hz,1H,4-H),4.07-3.89(m,9H,5-H,6-H,ArOCH 3 ),3.73(dd,J=7.9,6.7Hz,1H,2-H),1.79(t,J=5.8Hz,1H,OH),1.39(s,3H,CH 3 (isopropylidene)),1.31(s,3H,CH 3 (isopropylidene)),1.05(s,9H,C(CH 3 ) 3 (TBDPS)).

[0099] 13 C NMR (151 MHz, CDCl 3 )δ153.5,149.1,147.8,140.9,140.1,135.8,135.7,135.6,133.3,133.2,132.1,129.9,129.83,129.81,127.8,123 .5,118.9,111.3,110.6,108.1,101.0,79.9,78.8,74.6,73.3,70.5,63.7,63.0,56.5,56.4,27.8,26.9,26.3,19.3.

[0100] ESI-MS: found 827.31[M+Na] + .

[0101] (4) Synthesis of compound vi:

[0102]

[0103] Specific operations:

[0104] Under the protection of inert gas, compound v (60 mg, 0.074 mmol, 1.0 equiv.) and CA4 (23 mg, 0.074 mmol, 1.0 equiv.) were dissolved in a dry tetrahydrofuran solution (1 mL), the mixture was cooled to 0°C in an ice bath and stirred for 15 minutes, and tributylphosphine (20 μL, 0.081 mmol, 1.1 equiv.) and diisopropyl azodicarboxylate (15 μL, 0.077 mmol, 1.0 equiv.) were added in sequence, and the reaction solution was heated from 0°C to room temperature and continued to react for 1 hour. The reaction solution was diluted with dichloromethane and washed twice with saturated ammonium chloride, saturated sodium bicarbonate, and saturated sodium chloride solutions in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by reduced pressure distillation using a rotary evaporator. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate, 3:2, v / v) to give a light yellow oil vi (49 mg, 0.044 mmol, yield 60%).

[0105] 1 H NMR (600 MHz, CDCl 3 )δ7.80(s,1H,Ar),7.71-7.62(m,5H,Ar×4,Ar(DMNB)),7.45-7.29(m,8H,Ar×7,Ar(DMNB)),7.26-7.23(m,1H,Ar),6.90 (d,J=8.2Hz,1H,Ar(CA4)),6.84-6.77(m,2H,Ar(CA4)),6.48(s,2H,Ar(CA4)),6.43(s,2H,Ar(CA4)),5.25(s,2H,ArCH 2 (DMNB)),5.02(d,J=7.8Hz,1H,1-H),4.85(d,J=3.5Hz,2H,ArCH 2 ),4.30(t,J=6.2Hz,1H,3-H),4.24(d,J=5.8Hz,1H,4-H),4.01(t,J=6.61Hz,1H,5-H),3.97(s,3H,ArOCH 3 (DMNB)), 3.96-3.92(m,5H,ArOCH 3 (DMNB),6-H),3.84(s,3H,ArOCH 3 (CA4)),3.81(s,3H,ArOCH 3 (CA4)),3.72(t,J=7.4Hz,1H,2-H),3.67(s,6H,ArOCH 3 (CA4)),1.40(s,3H,CH 3(isopropylidene)),1.31(s,3H,CH 3 (isopropylidene)),1.04(s,9H,C(CH 3 ) 3 (TBDPS)).

[0106] 13 C NMR (101 MHz, CDCl 3 )δ153.4,153.1,149.4,149.0,147.7,147.1,140.8,140.0,135.7,135.6,133.2,1 33.1,132.9,132.6,132.0,129.91,129.89,129.88,129.8,129.5,129.1,127.8,12 4.0,123.2,118.6,114.9,111.6,111.2,110.5,108.0,106.0,100.9,79.8,78.7,74.4,73.2,70.5,69.4,62.9,60.9,56.5,56.4,55.99,55.97,27.9,26.8,26.3,19.2.

[0107] ESI-MS: found 1125.5[M+Na] + .

[0108] (5) Synthesis of Compound A:

[0109]

[0110] Specific operations:

[0111] Under argon protection, vi (40 mg, 0.036 mmol, 1.0 equiv.) was dissolved in a mixture of methanol and dichloromethane (1 mL, 4:1, v / v), and acetyl chloride (4.4 μL, 0.061 mmol, 2.0 equiv.) was added. The reaction solution was stirred at room temperature for 6 hours. After the reaction was completed, the reaction solution was directly concentrated, and the crude product was purified by silica gel column chromatography (methanol: dichloromethane, 1:20, v / v) to obtain a white solid compound A (16 mg, 0.019 mmol, yield 54%)

[0112] 1 H NMR (600 MHz, (CD 3 ) 2SO)δ7.85(d,J=2.1Hz,1H,Ar),7.64(s,1H,Ar(DMNB)),7.60(dd,J=8.8,2.2Hz,1H,Ar),7.46(d,J=9.5Hz,2H,Ar,Ar(DMNB)),6.95(d,J=1.9Hz,1H,Ar(CA4)),6.93(d,J=8.4Hz,1H,Ar(CA4)),6.89(dd,J=8.3,1.9Hz,1H,Ar(CA4)),6.56(s,2H,Ar(CA4)),6.51(d,J=12.2Hz,1H,Ar(CA4)),6.47(d,J=12.2Hz,1H,Ar(CA4)),5.38(d,J=7.6Hz,1H,1-H),5.14(d,J=15.7Hz,1H,ArCH 2 (DMNB)),5.06(d,J=15.6Hz,1H,ArCH 2 (DMNB)),4.90(s,2H,ArCH 2 ),3.83(s,3H,ArOCH 3 (CA4)),3.78(s,1H,4-H),3.76-3.72(m,7H,ArOCH 3 (DMNB),5-H),3.70-3.68(m,2H,3-H,2-H),3.63(s,3H,ArOCH 3 (CA4)),3.60(s,6H,ArOCH 3 (CA4)),3.57(dd,J=11.0,5.9Hz,1H,6-H),3.51(dd,J=10.9,6.6Hz,1H,6-H).

[0113] 13 C NMR(151MHz,(CD 3 ) 2 SO)δ153.3,152.6,148.9,148.6,147.1,147.0,139.3,138.5,136.8,133.4,132.4,131.0,130.7,129.3,129.2,128.7,123.9,122.2,116.9,114.5,112.0,110.2,107.8,106.0,100.1,79.9,75.8,72.4,70.5,68.5,68.1,60.04,60.02,56.0,55.8,55.63,55.61.

[0114] HRMS: m / z calculated [M+Na] + =847.2532, found 847.2536.

[0115] Example 2: Synthesis of Prodrug Compound B

[0116] (1) Synthesis of compound ix:

[0117]

[0118] Specific operations:

[0119] (a) To compound viii (600 mg, 1.84 mmol, 1.0 equiv.) were added toluene (20 mL) and dibutyltin oxide (550 mg, 2.21 mmol, 1.2 equiv.) in sequence under inert gas protection, and the solution was heated to reflux. After reflux for 6 hours, the mixture was returned to room temperature and the toluene was evaporated to dryness.

[0120] (b) The mixture after evaporation was dissolved in N,N-dimethylformamide (15 mL), and cesium fluoride (307 mg, 2.02 mmol, 1.1 equiv.), tetrabutylammonium bromide (652 mg, 2.02 mmol, 1.1 equiv.) and 4,5-dimethoxy-2-nitrobenzyl bromide (508 mg, 1.84 mmol, 1.0 equiv.) were added in sequence under the protection of inert gas. After stirring at 60°C for 16 hours, the N,N-dimethylformamide in the reaction solution was removed by reduced pressure distillation, and then dissolved in dichloromethane and washed twice with water, saturated ammonium chloride and saturated brine in sequence, and the organic phase was placed on anhydrous sodium sulfate for drying. After filtering off anhydrous sodium sulfate, the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 3:1, v / v) to obtain yellow solid compound ix (451 mg, 0.846 mmol, yield 46%).

[0121] 1 H NMR (400 MHz, CDCl 3 )δ7.69(s,1H,Ar(DMNB)),7.54(s,1H,Ar(DMNB)),7.52-7.45(m,2H,Ar),7.37-7.30(m,3H,Ar),5.52(s,1H,PhCH),5.19(d,J=15.9Hz,1H,ArCH 2 ),5.12(d,J=15.9Hz,1H,ArCH 2),4.45(d,J=9.7Hz,1H,1-H),4.41(dd,J=3.6,1.1Hz,1H,4-H),4.35(dd,J=12.5,1.6Hz,1H,6-H),4.12-4.01(m,2H,2-H,6-H),3.94(s,3H,ArOCH 3 ),3.74(s,3H,ArOCH 3 ),3.64(dd,J=9.2,3.5Hz,1H,3-H),3.49(q,J=1.5Hz,1H,5-H),3.32(hept,J=6.7Hz,1H,SCH),2.57(s,1H,OH),1.45-1.33(m,6H,CH 3 (isopropylthio)).

[0122] 13 C NMR (151 MHz, CDCl 3 )δ154.0,147.6,139.0,137.9,131.1,129.1,128.2,126.5,110.3,107.8,10 1.5,86.1,81.4,73.2,70.2,69.5,68.5,67.9,56.4,56.3,35.0,24.6,24.1.

[0123] ESI-MS: found 544.5[M+Na] + .

[0124] (2) Synthesis of compound x:

[0125]

[0126] Specific operations:

[0127] (a) Under inert gas protection, 80% acetic acid aqueous solution (5 mL, v / v) was added to compound ix (220 mg, 0.421 mmol, 1.0 equiv) and stirred at 60° C. for 12 hours. After the reaction, the reaction solution was dried by vacuum distillation to obtain a crude product after the benzylidene ketal was removed.

[0128] (b) After the crude product was co-evaporated with toluene twice, pyridine (5 mL), 4-dimethylaminopyridine (26 mg, 0.211 mmol, 0.5 equiv.) and acetic anhydride (0.18 mL, 1.90 mmol, 4.5 equiv.) were added to the dry crude product under inert gas protection at 0°C. The reaction was returned to room temperature and the reaction was continued for 12 hours. The reaction solution was diluted with EA and washed twice with water, 1M hydrochloric acid, saturated sodium bicarbonate and saturated brine in sequence. The organic phase was collected and dried over anhydrous sodium sulfate. After filtering off anhydrous sodium sulfate, the organic phase was concentrated and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate, 2:1, v / v) to obtain yellow foam compound x (201 mg, 0.358 mmol, yield 85%).

[0129] 1 H NMR (400 MHz, CDCl 3 )δ7.70(s,1H,Ar(DMNB)),7.18(s,1H,Ar(DMNB)),5.63(dd,J=3.4,1.1Hz,1H,4-H),5.25(t,J=9.8Hz,1H,2-H),5.10(dd,J=14.5,0.9Hz,1H,ArCH 2 ),4.91(dd,J=14.6,0.8Hz,1H,ArCH 2 ),4.58(d,J=10.1Hz,1H,1-H),4.17(qd,J=11.4,6.7Hz,2H,6-H),3.99(s,3H,ArOCH 3 ),3.94(s,3H,ArOCH 3 ),3.90(td,J=6.6,1.1Hz,1H,5-H),3.76(dd,J=9.4,3.4Hz,1H,3-H),3.20(hept,J=6.8Hz,1H,SCH),2.11(s,3H,CH 3 (Ac)), 2.07(s, 3H, CH 3 (Ac)), 2.06(s, 3H, CH 3 (Ac)), 1.44-1.33(m,6H,CH 3 (isopropylthio)).

[0130] 13 C NMR (101 MHz, CDCl 3)δ170.5,170.2,169.8,154.1,147.7,138.8,130.1,109.6,107.8,83.7,80. 0,74.5,69.4,66.7,61.9,56.7,56.4,35.6,24.1,23.8,21.1,20.81,20.77.

[0131] ESI-MS: found 582.6[M+Na] + .

[0132] (3) Synthesis of compound xi:

[0133]

[0134] Specific operations:

[0135] (a) Under inert gas protection, compound x (150 mg, 0.268 mmol, 1.0 equiv.) was dissolved in dry dichloromethane (3 mL). The mixture was cooled to 0°C in an ice bath and stirred for 10 minutes. Then, Molecular sieves and iodine monobromide (56 mg, 0.268 mmol, 1.0 equiv.). After the reaction is completed, the molecular sieves are filtered off, and the organic phase is collected and washed twice in sequence with 10% sodium thiosulfate, saturated sodium bicarbonate and saturated brine. The organic phase is then dried with anhydrous sodium sulfate, filtered, and then concentrated with a rotary evaporator under reduced pressure to obtain a crude bromide, which is directly used in the next step.

[0136] (b) The crude bromide was dissolved in dry acetonitrile (2.5 mL), and then 4-hydroxy-3-nitrobenzaldehyde (54 mg, 0.322 mmol, 1.2 equiv.), silver oxide (130 mg, 0.563 mmol, 2.1 equiv.), tetrabutylammonium iodide (50 mg, 0.134 mmol, 0.5 equiv.) and Molecular sieve. After 8 hours of reaction at room temperature, the silver oxide and molecular sieve were filtered using diatomaceous earth and the filter cake was washed with dichloromethane. The organic phase was washed twice in sequence with sodium carbonate, water, saturated sodium bicarbonate and saturated brine. The organic phase was collected and dehydrated with anhydrous sodium sulfate. After filtering out the sodium sulfate solid, the solvent was concentrated to obtain a crude mixture. The crude product was separated and purified using a silica gel column (petroleum ether: ethyl acetate, 2: 1, v / v) to obtain a light yellow solid compound xi (85 mg, 0.131 mmol, yield 49%).

[0137] 1 H NMR (600 MHz, CDCl 3)δ9.98 (singlet, 1H, ArCHO), 8.30 (doublet, J = 1.9 Hz, 1H, Ar), 8.07 (doublet of doublets, J = 8.6, 1.9 Hz, 1H, Ar), 7.71 (singlet, 1H, Ar(DMNB)), 7.50 (doublet, J = 8.6 Hz, 1H, Ar), 7.20 (singlet, 1H, Ar(DMNB)), 5.68 (doublet, J = 3.3 Hz, 1H, 4-H), 5.61 (doublet of doublets, J = 9.9, 7.9 Hz, 1H, 2-H), 5.22 (doublet, J = 7.9 Hz, 1H, 1-H), 5.14 (doublet, J = 14.4 Hz, 1H, ArCH 2 (DMNB)), 4.92 (doublet, J = 14.3 Hz, 1H, ArCH 2 (DMNB)), 4.29 (doublet of doublets, J = 11.6, 7.0 Hz, 1H, 6-H), 4.23 (doublet of doublets, J = 11.6, 6.0 Hz, 1H, 6-H), 4.13 (triplet, J = 6.6 Hz, 1H, 5-H), 4.00 (singlet, 3H, ArOCH 3 ), 3.95 (singlet, 3H, ArOCH 3 ), 3.86 (doublet of doublets, J = 10.2, 3.4 Hz, 1H, 3-H), 2.15 (singlet, 3H, CH 3 (Ac)), 2.11 (singlet, 6H, CH 3 (Ac)).

[0138] 13 C NMR (151 MHz, CDCl 3 ) δ 188.6, 170.4, 170.0, 169.5, 154.2, 153.7, 147.9, 138.8, 133.9, 131.5, 129.7, 126.9, 118.8, 109.6, 108.0, 100.1, 78.5, 72.0, 69.5, 65.9, 61.7, 56.7, 56.4, 20.8, 20.73, 20.68.

[0139] ESI-MS: found 673.2 [M+Na] + .

[0140] (4) Synthesis of compound xii:

[0141]

[0142] Specific operation:

[0143] Under the protection of inert gas, compound xi (80 mg, 0.123 mmol, 1.0 equiv.) was dissolved in a mixed solution of dichloromethane and isopropanol (2 mL), the mixed solution was cooled to 0°C in an ice bath, sodium borohydride powder (9 mg, 0.246 mmol, 2.0 equiv.) was added, the reaction solution was slowly heated from 0°C to room temperature and continued to react for 2 hours. After TLC confirmed that the crude raw material reaction was complete. Then the reaction solution was diluted with dichloromethane, and washed twice in sequence with water, saturated ammonium chloride, saturated sodium bicarbonate, and saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, and the anhydrous sodium sulfate was filtered off and the organic phase was concentrated by vacuum distillation using a rotary evaporator. After concentration, the crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 1:1, v / v) to obtain a light yellow oily compound xii (74 mg, 0.114 mmol, yield 93%).

[0144] 1 H NMR (600 MHz, CDCl 3 )δ7.81(d,J=2.1Hz,1H,Ar),7.71(s,1H,Ar(DMNB)),7.53(dd,J=8.7,2.1Hz,1H,Ar),7.37(d,J=8.6Hz,1H,Ar), 7.21(s,1H,Ar(DMNB)),5.66(d,J=3.3Hz,1H,4-H),5.57(dd,J=9.9,8.0Hz,1H,2-H),5.14(d,J=14.6Hz,1H,ArCH 2 (DMNB)),5.07(d,J=8.0Hz,1H,1-H),4.91(d,J=14.5Hz,1H,ArCH 2 (DMNB)),4.73(s,2H,ArCH 2 ),4.29(dd,J=11.4,6.9Hz,1H,6-H),4.21(dd,J=11.4,6.2Hz,1H,6-H),4.05(t,J=6.6Hz,1H,5-H),4.00(s,3H,ArOCH 3 ),3.95(s,3H,ArOCH 3 ),3.83(dd,J=10.0,3.3Hz,1H,3-H),2.18-2.09(m,9H,CH 3 (Ac)).

[0145] 13 C NMR (151 MHz, CDCl 3)δ170.4,170.1,169.7,154.2,148.7,147.8,141.5,138.8,137.0,131.7,129.9,123.3,120.0 ,109.6,107.9,100.9,78.7,71.6,69.8,69.4,66.1,63.6,61.7,56.7,56.4,20.9,20.8,20.7.

[0146] ESI-MS: found 675.5[M+Na] + .

[0147] (5) Synthesis of Compound xiii:

[0148]

[0149] Specific operations:

[0150] Under inert gas protection, compound xii (35 mg, 0.054 mmol, 1.0 equiv.) and CA4 (17 mg, 0.054 mmol, 1.0 equiv.) were dissolved in a dry tetrahydrofuran solution (1 mL), the mixture was cooled to 0°C in an ice bath and stirred for 15 minutes, and tributylphosphine (15 μL, 0.060 mmol, 1.1 equiv.) and diisopropyl azodicarboxylate (11 μL, 0.054 mmol, 1.0 equiv.) were added in sequence, and the reaction solution was heated from 0°C to room temperature and continued to react for 1 hour. The reaction solution was diluted with dichloromethane and washed twice with saturated ammonium chloride, saturated sodium bicarbonate, and saturated sodium chloride solutions in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by reduced pressure distillation using a rotary evaporator. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate, 3:2, v / v) to give a light yellow oil xiii (42 mg, 0.044 mmol, yield 83%).

[0151] 1 H NMR (400 MHz, CDCl 3)δ7.80(d,J=2.3Hz,1H,Ar),7.72(s,1H,Ar(DMNB)),7.44(dd,J=8.6,2.2Hz,1H,Ar),7.32(d,J=8.6Hz,1H,Ar),7.22(s,1H,Ar(DMNB)),6.91(dd,J=8.3,2.0Hz,1H,Ar(CA4)),6.81(m,J=8.6Hz,2H,Ar(CA4)),6.50-6.42(m,4H,Ar(CA4)),5.66(dd,J=3.4,1.1Hz,1H,4-H),5.57(dd,J=10.0,8.0Hz,1H,2-H),5.15(d,J=15.1Hz,1H,ArCH 2 (DMNB)),5.09(d,J=8.0Hz,1H,1-H),4.96-4.87(m,3H,ArCH 2 (DMNB),ArCH 2 ),4.25(qd,J=11.3,6.4Hz,2H,6-H),4.07(t,J=6.7Hz,1H,5-H),4.00(s,3H,ArOCH 3 (DMNB)),3.95(s,3H,ArOCH 3 (DMNB)),3.87(s,3H,ArOCH 3 (CA4)),3.88-3.81(m,4H,3-H,ArOCH 3 (CA4)),3.69(s,6H,ArOCH 3 (CA4)),2.15-2.10(m,9H,CH 3 (Ac)).

[0152] 13 C NMR(101MHz,CDCl 3 )δ170.5,170.1,169.7,154.2,153.1,149.02,149.01,147.7,147.1,141.3,138.7,133.1,133.0,132.5,132.2,130.0,129.9,129.5,129.1,125.1,123.8,119.7,114.8,111.6,109.5,107.8,105.9,100.7,100.6,78.7,71.6,69.7,69.5,66.1,62.3,61.0,56.6,56.3,56.0,55.9,20.9,20.8,20.7.

[0153] ESI-MS: found 973.5[M+Na] + .

[0154] (6) Synthesis of Compound B:

[0155]

[0156] Specific operations:

[0157] Under inert gas protection, compound xiii (42 mg, 0.044 mmol, 1.0 equiv.) was dissolved in methanol (1 mL), followed by the addition of 5 mol / L sodium methoxide methanol solution (5 μL, 0.025 mmol, 0.6 equiv.) and stirring at room temperature for 2 hours. The reaction was terminated when TLC detected that all the raw materials had reacted and a single main point was generated. H + The reaction solution was neutralized with ion exchange resin, and after the pH of the solution was detected to be neutral to slightly acidic, the resin was filtered off and washed with a mixture of methanol and methanol-dichloromethane (2:1, v / v). The crude product was concentrated by vacuum distillation using a rotary evaporator, and the crude product was purified using a silica gel column (methanol: dichloromethane, 1:25, v / v) to obtain a white solid compound B (24 mg, 0.030 mmol, yield 68%).

[0158] 1 H NMR (600 MHz, (CD 3 ) 2 SO)δ7.86(d,J=2.2Hz,1H,Ar),7.80(s,1H,Ar(DMNB)),7.69(s,1H,Ar(DMNB)),7.58(dd,J=8. 8,2.2Hz,1H,Ar),7.43(d,J=8.8Hz,1H,Ar),6.96(d,J=2.0Hz,1H,Ar(CA4)),6.93(d,J=8.4Hz ,1H,Ar(CA4)),6.89(dd,J=8.4,1.9Hz,1H,Ar(CA4)),6.56(s,2H,Ar(CA4)),6.53-6.44(m,2H ,Ar(CA4)),5.75(d,J=7.6Hz,1H,OH),5.13(d,J=7.7Hz,1H,1-H),5.06(d,J=16.4Hz,1H,ArCH 2 ),4.95(d,J=16.4Hz,1H,ArCH 2 ),4.93-4.86(m,3H,ArCH 2(DMNB),OH),4.74(t,J=5.4Hz,1H,OH),4.12-4.03(m,1H,4-H),3.92(s,3H,ArOCH 3 (DMNB)), 3.89-3.82(m,4H,ArOCH 3 (DMNB), 2-H), 3.74(s, 3H, ArOCH 3 (CA4)), 3.66-3.61(m,4H,ArOCH 3 (CA4),5-H),3.61-3.57(m,7H,ArOCH 3 (CA4)×2,6-H),3.52(dt,J=11.1,5.5Hz,1H,6-H),3.43(dd,J=9.6,3.1Hz,1H,3-H).

[0159] 13 C NMR (151 MHz, (CD 3 ) 2 SO)δ153.8,152.6,149.2,148.6,147.1,147.0,139.8,138.6,136.7,133.3,132.4,131.5,130.7,129.3,128.8,123.9,122.2, 117.1,114.4,112.0,110.4,107.8,105.9,100.7,81.4,75.5,68.8,68.5,66.0,63.8,60.09,60.06,56.3,56.0,55.64,55.60.

[0160] HRMS: m / z calculated [M+Na] + =847.2532, found 847.2536.

[0161] Example 3: Synthesis of Prodrug Compound C

[0162] (1) Synthesis of compound xvi:

[0163]

[0164] Specific operations:

[0165] Toluene (7 mL) and dibutyltin oxide (225 mg, 0.901 mmol, 1.2 equiv.) were added to compound xv (200 mg, 0.751 mmol, 1.0 equiv.) in sequence under inert gas protection, and the solution was heated to reflux. After 6 hours of reflux, the mixture was returned to room temperature and the toluene was evaporated. The evaporated mixture was dissolved in N, N-dimethylformamide (7 mL), and tetrabutylammonium bromide (266 mg, 0.826 mmol, 1.1 equiv.) and 4,5-dimethoxy-2-nitrobenzyl bromide (207 mg, 0.751 mmol, 1.0 equiv.) were added in sequence under inert gas protection. After stirring at 60°C for 12 hours, N, N-dimethylformamide in the reaction solution was removed by reduced pressure distillation, and then diluted with dichloromethane and washed twice with water, saturated ammonium chloride and saturated brine in sequence, and the organic phase was dried over anhydrous sodium sulfate. After filtering off anhydrous sodium sulfate, the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate, 7:3, v / v) to obtain yellow oily compound xvi (194 mg, 0.421 mmol, yield 56%).

[0166] 1 H NMR (400 MHz, CDCl 3 )δ8.08-7.98(m,2H,Ar),7.69(s,1H,Ar(DMNB)),7.63-7.53(m,1H,Ar),7.45(t,J=7.8Hz,2H,Ar), 7.18(s,1H,Ar(DMNB)),5.57(t,J=1.5Hz,1H,1-H),5.14-5.12(m,1H,2-H),5.09-5.07(m,2H,ArCH 2 ),4.66(td,J=4.6,1.3Hz,1H,5-H),4.45(d,J=7.4Hz,1H,6-H),4.32-4.26(m,1H,3-H),4.01(t,J=4.6Hz,1H,4-H),3.98(s,3H,ArOCH 3 ),3.95(s,3H,ArOCH 3 ),3.76(ddd,J=7.4,5.2,1.0Hz,1H,6-H),2.83(s,1H,OH).

[0167] ESI-MS: found 484.1[M+Na] + .

[0168] (2) Synthesis of compound xvii:

[0169]

[0170] Specific operations:

[0171] Under the protection of inert gas at 0°C, compound xvi was dissolved in acetic anhydride solution (2 mL) containing 2% concentrated sulfuric acid. After the reaction was restored to room temperature for 2 hours, TLC detected that the reaction was complete, and the mixture was poured into stirred ice water and neutralized with solid sodium bicarbonate, and extracted with dichloromethane 2-3 times. The dichloromethane phase was then washed twice with water, saturated sodium bicarbonate and saturated brine in sequence, and dried over anhydrous sodium sulfate. After filtering off anhydrous sodium sulfate, the organic phase was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 1: 1, v / v) to obtain yellow foam compound xvii (165 mg, 0.273 mmol, yield 84%, α: β = 5: 1)

[0172] ESI-MS: found 628.2[M+Na] + .

[0173] (3) Synthesis of compound xviii:

[0174]

[0175] Specific operations:

[0176] (a) Compound xvii (200 mg, 0.330 mmol, 1.0 equiv.) was dissolved in dichloromethane (3 mL) at 0°C under inert gas protection. 33% hydrobromic acid acetic acid solution (0.24 mL, 2.31 mmol, 7.0 equiv.) was added dropwise to the mixture at 0°C under inert gas protection. The mixture was allowed to react for 2 hours at room temperature. When TLC showed that the reaction of the raw material was complete, the reaction solution was diluted with dichloromethane and washed twice with saturated sodium bicarbonate and saturated sodium chloride solutions in sequence. The organic phase was collected, dried over anhydrous sodium sulfate, the anhydrous sodium sulfate was filtered, and concentrated by reduced pressure distillation using a rotary evaporator to obtain a crude bromide.

[0177] (b) Under inert gas protection, the crude bromide was dissolved in acetonitrile, and then 4-hydroxy-3-nitrobenzaldehyde (66 mg, 0.396 mmol, 1.2 equiv.), silver oxide (162 mg, 0.759 mmol, 2.3 equiv.), tetrabutylammonium iodide (61 mg, 0.165 mmol, 0.5 equiv.) and Molecular sieve. After reacting at room temperature for 12 hours, the silver oxide and molecular sieve were filtered using diatomaceous earth and the filter cake was washed with dichloromethane. The organic phase was washed twice with sodium carbonate, water and saturated brine in sequence. The organic phase was collected and dehydrated with anhydrous sodium sulfate. After filtering out the sodium sulfate solid, the solvent was concentrated to obtain a crude mixture. The crude product was separated and purified using a silica gel column (petroleum ether: ethyl acetate, 2: 1, v / v) to obtain a light yellow oily compound xviii (111 mg, 0.155 mmol, yield 47%).

[0178] 1 H NMR (600 MHz, CDCl 3 )δ9.94(s,1H,ArCHO),8.24(d,J=2.0Hz,1H,Ar),8.05(dd,J=8.7,2.0Hz,1H,A r),8.03-7.98(m,2H,Ar),7.73(s,1H,Ar(DMNB)),7.60-7.55(m,1H,Ar),7.52 -7.41(m,4H,Ar(DMNB),Ar×3),6.02(dd,J=10.2,7.5Hz,1H,2-H),5.43(d,J=7 .5Hz,1H,1-H),5.39(dd,J=10.2,2.9Hz,1H,3-H),5.23(d,J=14.9Hz,1H,ArCH 2 ),5.13(d,J=15.1Hz,1H,ArCH 2 ),4.52(dd,J=11.4,7.1Hz,1H,6-H),4.28(dd,J=11.4,5.9Hz,1H,6-H),4.24(dd, J=3.0,1.4Hz,1H,4-H),4.19(ddd,J=7.2,5.9,1.4Hz,1H,5-H),4.12(s,3H,ArOCH 3 ),3.97(s,3H,ArOCH 3 ),2.10(s,3H,CH 3 (Ac)),1.97(s,3H,CH 3 (Ac)).

[0179] 13 C NMR (151 MHz, CDCl 3)δ188.6,170.4,170.2,164.9,154.3,153.5,147.9,141.2,138.6,133.9,133.5,131.3,130.0,129.9,12 9.1,128.5,127.0,118.4,109.6,107.9,99.7,75.5,73.2,72.5,72.2,68.8,62.1,56.8,56.4,20.8,20.7.

[0180] ESI-MS: found 735.2[M+Na] + .

[0181] (4) Synthesis of compound xix:

[0182]

[0183] Specific operations:

[0184] Under the protection of inert gas, compound xviii (100 mg, 0.140 mmol, 1.0 equiv.) was dissolved in a mixed solution of dichloromethane and isopropanol (2 mL), the mixed solution was cooled to 0°C in an ice bath, sodium borohydride powder (11 mg, 0.280 mmol, 2.0 equiv.) was added, the reaction solution was slowly heated from 0°C to room temperature and the reaction was continued for 2 hours. After TLC confirmed that the crude raw material had completed the reaction. Then the reaction solution was diluted with dichloromethane, and washed twice in sequence with water, saturated ammonium chloride, saturated sodium bicarbonate, and saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, and the anhydrous sodium sulfate was filtered off and the organic phase was concentrated by vacuum distillation using a rotary evaporator. After concentration, the crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 3:2, v / v) to obtain a light yellow oil xix (89 mg, 0.125 mmol, yield 89%).

[0185] 1 H NMR (600 MHz, CDCl 3<h2 style=";text-align:left;direction:ltr">(δ8.02(dd,J=8.2,1.4Hz,2H,Ar),7.74-7.69(m,2H,Ar(DMNB),Ar),7.59-7.54) m,1H,Ar),7.54(s,1H,Ar(DMNB)),7.50(dd,J=8.7,2.2Hz,1H,Ar),7.43(t,J=7.8 Hz,2H,Ar),7.31(d,J=8.6Hz,1H,Ar),5.98(dd,J=10.4,7.7Hz,1H,2-H),5.34(dd ,J=10.4,2.9Hz,1H,3-H),5.27(d,J=7.7Hz,1H,1-H),5.22(d,J=15.2Hz,1H,ArCH<h2 style=";text-align:left;direction:ltr"> 2 <h2 style=";text-align:left;direction:ltr"> ),5.11(d,J=15.3Hz,1H,ArCH<h2 style=";text-align:left;direction:ltr"> 2 <h2 style=";text-align:left;direction:ltr"> ),4.68(s,2H,ArCH<h2 style=";text-align:left;direction:ltr"> 2 <h2 style=";text-align:left;direction:ltr"> (DMNB)),4.50(dd,J=11.3,6.9Hz,1H,6-H),4.25(dd,J=11.3,6.3Hz,1H,6-H),4.20(dd,J=3.0,1.2Hz,1H,4-H),4.13(s,3H,ArOCH<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> ),4.09(td,J=7.9,7.2,1.9Hz,1H,5-H),3.97(s,3H,ArOCH<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> ),2.09(s,3H,CH<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> (Ac)),1.95(s,3H,CH<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> (Ac)).<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0186] <h2 style=";text-align:left;direction:ltr"> 13 <h2 style=";text-align:left;direction:ltr"> C NMR (151MHz, CDCl<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> )δ170.4,170.3,165.1,154.4,148.4,147.8,141.5,138.4,136.9,133.4,131.7,130.3,129.9,129.3,12 8.5,123.4,119.6,109.6,107.8,100.6,75.7,72.9,72.8,72.2,69.1,63.5,62.1,56.9,56.4,20.8,20.7.<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0187] <h2 style=";text-align:left;direction:ltr"> ESI-MS:found 737.2[M+Na]<h2 style=";text-align:left;direction:ltr"> + <h2 style=";text-align:left;direction:ltr"> .<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0188] <h2 style=";text-align:left;direction:ltr"> (5)More<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0189] <h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0190] Specific operations:

[0191] Under the protection of inert gas, compound xix (35 mg, 0.049 mmol, 1.0 equiv.) and CA4 (15 mg, 0.049 mmol, 1.0 equiv.) were dissolved in a dry tetrahydrofuran solution (1 mL), the mixture was cooled to 0°C in an ice bath and stirred for 15 minutes, and tributylphosphine (13 μL, 0.054 mmol, 1.1 equiv.) and diisopropyl azodicarboxylate (10 μL, 0.049 mmol, 1.0 equiv.) were added in sequence, and the reaction solution was heated from 0°C to room temperature and continued to react for 1 hour. The reaction solution was diluted with dichloromethane and washed twice with saturated ammonium chloride, saturated sodium bicarbonate, and saturated sodium chloride solutions in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by reduced pressure distillation using a rotary evaporator. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate, 3:2, v / v) to give a light yellow oily compound xx (35 mg, 0.034 mmol, yield 70%).

[0192] 1 H NMR (600 MHz, CDCl 3 )δ8.06-8.00(m,2H,Ar),7.75-7.71(m,2H,Ar(DMNB),Ar),7.59-7.54(m,2H,Ar(DMNB),A r),7.47-7.38(m,3H,Ar),7.29-7.26(m,1H,Ar),6.89(dd,J=8.3,2.0Hz,1H,Ar(CA4)),6. 82-6.76(m,2H,Ar(CA4)),6.51-6.38(m,4H,Ar(CA4)),5.99(dd,J=10.4,7.7Hz,1H,2-H), 5.37(dd,J=10.4,3.0Hz,1H,3-H),5.33(d,J=7.7Hz,1H,1-H),5.23(d,J=15.3Hz,1H,ArCH 2 (DMNB)),5.12(d,J=15.3Hz,1H,ArCH 2 (DMNB)),4.87(s,2H,ArCH 2 ),4.49(dd,J=11.2,6.9Hz,1H,6-H),4.27(dd,J=11.3,6.3Hz,1H,6-H),4.22(dd,J=3.0,1.3Hz,1H,4-H),4.14(s,3H,ArOCH 3 (DMNB)),3.97(s,3H,ArOCH 3(DMNB)),3.84(s,3H,ArOCH 3 (CA4)),3.82(s,3H,ArOCH 3 (CA4)),3.65(s,6H,ArOCH 3 (CA4)), 2.10(d, J = 1.2 Hz, 3H, CH 3 (Ac)), 1.96(d, J=1.1Hz, 3H, CH 3 (Ac)).

[0193] ESI-MS: found 1035.7[M+Na] + .

[0194] (6) Synthesis of Compound C:

[0195]

[0196] Specific operations:

[0197] Under inert gas protection, compound xx (35 mg, 0.034 mmol, 1.0 equiv.) was dissolved in a mixture of methanol and dichloromethane (1 mL), followed by the addition of 5 mol / L sodium methoxide methanol solution (4 μL, 0.021 mmol, 0.6 equiv.) and stirring at room temperature for 2 hours. The reaction was terminated when all the raw materials were reacted and a single main point was generated by TLC. H + The reaction solution was neutralized with ion exchange resin, and after the pH of the solution was detected to be neutral to slightly acidic, the resin was filtered off and washed with a mixture of methanol and methanol-dichloromethane (2:1, v / v). The crude product was concentrated by vacuum distillation using a rotary evaporator, and the crude product was purified using a silica gel column (methanol: dichloromethane, 1:25, v / v) to obtain a white solid compound C (23 mg, 0.028 mmol, yield 81%).

[0198] 1 H NMR (600 MHz, (CD 3 ) 2SO)δ7.85(d,J=2.1Hz,1H,Ar),7.67(s,1H,Ar(DMNB)),7.61-7.56(m,2H,Ar,Ar(DMNB)),7.45(d,J=8.8Hz,1H,Ar),6.96(d,J=1.9Hz,1H,Ar(CA4)),6.93(d,J=8.3Hz,1H,Ar(CA4)),6.89(dd,J=8.3,1.9Hz,1H,Ar(CA4)),6.56(s,2H,Ar(CA4)),6.49(q,J=12.2Hz,2H,Ar(CA4)),5.39(d,J=5.2Hz,1H,OH),5.35(d,J=4.8Hz,1H,OH),5.21(d,J=15.4Hz,1H,ArCH 2 (DMNB)),5.15(d,J=7.5Hz,1H,1-H),5.00(d,J=15.5Hz,1H,ArCH 2 (DMNB)),4.91(s,2H,ArCH 2 ),4.86(t,J=5.4Hz,1H,OH),3.91(s,3H,ArOCH 3 (DMNB)),3.88-3.85(m,4H,ArOCH 3 (DMNB),4-H),3.78(t,J=6.7Hz,1H,5-H),3.74(s,3H,ArOCH 3 ),3.69(ddd,J=9.7,7.5,5.2Hz,1H,2-H),3.64-3.59(m,10H,3-H,ArOCH 3 ×3),3.52(ddd,J=10.8,7.1,4.4Hz,2H,6-H).

[0199] 13 C NMR(151MHz,(CD 3 ) 2 SO)δ153.4,152.6,149.1,148.6,147.1,147.0,140.1,138.5,136.7,133.3,132.4,131.3,130.9,129.3,128.8,123.8,122.2,117.3,114.4,112.0,110.4,107.7,105.9,100.8,77.3,75.3,73.7,70.6,68.6,60.1,59.7,56.02,55.98,55.64,55.60,54.9.

[0200] HRMS: m / z calculated [M+Na] + =847.2532, found 847.2540.

[0201] Example 4: Synthesis of Prodrug D

[0202] (1) Synthesis of compound xxiii:

[0203]

[0204] Specific operations:

[0205] Under the protection of inert gas, compound xxii (4.01 g, 8.04 mol, 1.0 equiv.) was dissolved in N, N-dimethylformamide, and then imidazole (1093 mg, 16.08 mol, 2.0 equiv.) was added to the mixed solution system, and the mixed solution was ice-bathed to 0°C, and then TBDPSCl (3.13 mL, 12.06 mol, 1.5 equiv.) was added dropwise to the solution system. After the addition was completed, the reaction solution was raised to room temperature and reacted for 2 hours. TLC detected that the raw materials were completely reacted to generate a main product point, and the reaction solution was cooled to 0°C and sodium bicarbonate solution was added to the reaction solution system to terminate the reaction. The mixture was diluted with dichloromethane, washed twice with saturated sodium bicarbonate and saturated sodium chloride solutions in sequence, the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated by reduced pressure distillation on a rotary evaporator. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 10: 1, v / v) to give white foam xxiii (5.63 g, 7.64 mmol, yield 95%).

[0206] 1 H NMR (600 MHz, CDCl 3 )δ7.72(d,J=2.1Hz,1H,Ar),7.68-7.60(m,4H,Ar),7.50-7.41(m,3H,Ar),7.39(td,J=7.2,1.5Hz,4H,Ar),7.32(d,J=8.6Hz,1H,Ar),5.55(d d,J=10.5,7.9Hz,1H,2-H),5.47(dd,J=3.4,1.1Hz,1H,4-H),5.11(dd,J=10.5,3.4Hz,1H,3-H),5.05(d,J=7.9Hz,1H,1-H),4.73(s,2H,ArCH 2 ),4.27(dd,J=11.4,7.0Hz,1H,6-H),4.17(dd,J=11.4,6.3Hz,1H,6-H),4.10-3.99(m,1H,5-H),2.19(s,3H,CH3 (Ac)),2.14(s,3H,CH 3 (Ac)), 2.06(s, 3H, CH 3 (Ac)),2.02(s,3H,CH 3 (Ac)),1.10(s,9H,C(CH 3 ) 3 (TBDPS)).

[0207] 13 C NMR (151 MHz, CDCl 3 )δ170.3,170.2,170.2,169.5,148.1,141.3,137.4,135.5,132.8,131.0,130.0,127.9,122 .6,119.9,100.9,71.4,70.6,67.9,66.7,64.1,61.3,26.8,20.7,20.66,20.65,20.6,19.3.

[0208] ESI-MS: found 760.2[M+Na] + .

[0209] (2) Synthesis of compound xxiii:

[0210]

[0211] Specific operations:

[0212] (a) Under inert gas protection, compound xxiii (5.5 g, 7.45 mmol, 1.0 equiv.) was dissolved in methanol (40 mL), followed by the addition of 5 mol / L sodium methoxide methanol solution (745 μL, 3.73 mmol, 0.5 equiv.) and stirring at room temperature for 2 hours. The reaction was terminated when all the raw materials were reacted and only one main point was generated according to TLC. H + The reaction solution was neutralized with ion exchange resin, and after the pH of the solution was tested to be neutral to slightly acidic, the resin was filtered off and washed with a mixture of methanol and methanol-dichloromethane (2:1, v / v). The solution was concentrated by vacuum distillation using a rotary evaporator to obtain a crude product of 3-nitrobenzyl tert-butyl diphenyl chlorosilyl ether β-D-galactopyranoside, which was directly used in the next step.

[0213] (b) The crude product was dissolved in dry pyridine. After the reactants were fully dissolved, the reaction system was cooled to 0°C and stirred for 15 minutes. Triphenylmethane (6.23 g, 22.35 mmol, 3.0 equiv.) was then added. After stirring at room temperature for 1 hour, 1.5 mL of triethylamine was added dropwise to the solution. After stirring at room temperature for 5 hours, TLC showed no significant changes in the reactants and products, and the reaction was terminated. The reaction solution was diluted with dichloromethane, washed twice with 1 mol / L hydrochloric acid, saturated sodium bicarbonate, and saturated saline solution in sequence, the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated by reduced pressure distillation on a rotary evaporator. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate: methanol, 1:2:0.05, v / v / v) to finally obtain a transparent flaky solid xxiv (3.75 g, 4.62 mmol, yield 62%).

[0214] 1 H NMR (600 MHz, (CD 3 ) 2 SO)δ7.80(d,J=2.0Hz,1H,Ar),7.68-7.56(m,6H,Ar),7.48-7.32(m,12H,Ar),7.26(t,J=7.7Hz,6H,Ar),7.21-7 .14(m,3H,Ar),5.24(d,J=5.2Hz,1H,OH),5.17(d,J=7.7Hz,1H,1-H),4.95(d,J=5.8Hz,1H,OH),4.76(s,2H,ArCH 2 ),4.63(d,J=4.6Hz,1H,OH),3.98(dd,J=8.3,3.2Hz,1H,5-H),3.62-3.53(m,2H,4-H,6-H),3.43(ddd,J=9. 3,5.7,3.2Hz,1H,6-H),3.29(dd,J=9.8,8.1Hz,1H,2-H),2.96(dd,J=9.9,3.1Hz,1H,3-H),1.00(s,9H,C(CH 3 ) 3 (TBDPS)).

[0215] 13 C NMR (151 MHz, (CD 3 ) 2SO)δ148.6,143.8,139.8,134.98,134.96,134.3,132.6,132.5,131.0,130.0,128.3,128. 0,127.8,126.9,122.0,117.0,100.6,85.8,74.5,73.2,69.9,68.9,63.9,63.7,26.6,18.8.

[0216] ESI-MS: found 834.6[M+Na] + .

[0217] (3) Synthesis of compound xxv:

[0218]

[0219] Specific operation: Under the protection of inert gas, compound xxiv (3.60 g, 4.43 mmol, 1.0 equiv.) was dissolved in pyridine (25 mL), and the reaction system was cooled to 0°C, and then benzoyl chloride (2.32 mL, 20 mmol, 4.5 equiv.) was added dropwise to the solution system. After the addition was complete, the solution was returned to room temperature and stirred for 6 hours. TLC detected that all the raw materials were consumed, and the reaction was terminated. The mixture was concentrated by reduced pressure distillation using a rotary evaporator, and the crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 1:1.5, v / v) to finally obtain white foam xxv (yield 85%).

[0220] 1 HNMR (400MHz, CDCl 3 )δ8.12(ddd,J=11.5,8.4,1.4Hz,2H,Ar),8.04-7.96(m,2H,Ar),7.86-7.7 7(m,2H,Ar),7.68-7.57(m,6H,Ar),7.56-7.30(m,14H,Ar),7.28(d,J=8.0 Hz,2H,Ar),6.17(dd,J=10.4,7.9Hz,1H,2-H),5.95-5.86(m,1H,4-H),5.6 8(dd,J=10.5,3.4Hz,1H,3-H),5.42(d,J=7.9Hz,1H,1-H),4.71(s,2H,ArCH 2 ),4.23-4.16(m,1H,5-H),3.89(dd,J=12.0,6.7Hz,1H,6-H),3.72(dd,J=12.0,6.5Hz,1H,6-H),1.08(s,9H,C(CH 3 ) 3 (TBDPS)).

[0221] 13 C NMR (151 MHz, CDCl 3 )δ166.7,165.6,165.2,148.0,141.5,137.3,135.5,134.0,133.5,133.2,132.9,131.2,130.2,130.1,130.0,129.83,129.82,12 9.3,128.8,128.59,128.56,128.44,128.40,128.3,127.9,122.7,119.4,101.1,74.8,71.6,69.1,68.5,64.1,60.6,26.8,19.3.

[0222] ESI-MS: found 904.3[M+Na] + .

[0223] (4) Synthesis of compound xxvii:

[0224]

[0225] Specific operation: Compound xxv (100 mg, 0.113 mmol, 1.0 equiv.) and xxvi (61 mg, 0.170 mmol, 1.5 equiv.) were dissolved in dichloromethane (1 mL) under inert gas protection. The reaction system was cooled to 0°C and trifluoromethanesulfonic acid (5 μL, 0.056 mmol, 0.5 equiv.) was added to the solution system using a microsyringe. The temperature was slowly raised to room temperature after the addition. TLC detected that the raw material of compound xxvi was completely consumed and a new spot was generated. The temperature was cooled to 0°C and saturated sodium bicarbonate was added to terminate the reaction. The reaction solution was diluted with dichloromethane, and washed twice with saturated sodium bicarbonate and saturated saline solution in sequence. The organic phase was collected and dried with anhydrous sodium sulfate, filtered, and then concentrated by reduced pressure distillation using a rotary evaporator. The crude product was separated and purified by silica gel column chromatography (petroleum ether: ethyl acetate, 3:2, v / v) to finally obtain a light yellow transparent oil xxvii (54.8 mg, 0.051 mmol, yield 45%).

[0226] 1 H NMR (400 MHz, CDCl 3<h2 style=";text-align:left;direction:ltr">)δ8.10-7.93(m,4H,Ar),7.85-7.77(m,2H,Ar),7.72-7.56(m,7H,Ar×6,Ar(DMNB)),7.54-7.34(m,14H,Ar),7.28-7.24(m,3H,Ar ×2,Ar(DMNB)),6.18-6.05(m,2H,2-H,4-H),5.66(dd,J=10.5,3.4Hz,1H,3-H),5.46(d,J=8.0Hz,1H,1-H),5.01-4.85(m,2H,ArCH<h2 style=";text-align:left;direction:ltr"> 2 <h2 style=";text-align:left;direction:ltr"> ),4.71(s,2H,ArCH<h2 style=";text-align:left;direction:ltr"> 2 <h2 style=";text-align:left;direction:ltr"> '),4.49-4.36(m,1H,5-H),4.03-3.95(m,2H,6-H),3.91(d,J=6.5Hz,7H,ArOCH<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> ,6-H),1.08(s,9H,C(CH<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> )<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> (TBDPS)).<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0227] <h2 style=";text-align:left;direction:ltr"> 13 <h2 style=";text-align:left;direction:ltr"> C NMR (101MHz, CDCl<h2 style=";text-align:left;direction:ltr"> 3 <h2 style=";text-align:left;direction:ltr"> )δ165.7,165.5,165.2,154.1,148.1,147.7,141.3,139.0,137.2,135.5 ,133.7,133.4,133.2,132.9,131.1,130.0,129.9,129.84,129.79,129. 4,128.9,128.7,128.4,128.3,127.9,122.8,119.3,109.7,107.8,101.1 ,73.1,71.7,70.3,69.0,68.5,67.9,64.1,56.6,56.4,56.3,26.8,19.3.<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0228] <h2 style=";text-align:left;direction:ltr"> ESI-MS:found 1099.3[M+Na]<h2 style=";text-align:left;direction:ltr"> + <h2 style=";text-align:left;direction:ltr"> .<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0229] <h2 style=";text-align:left;direction:ltr"> (5)More<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0230] <h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr">

[0231] Specific operation: Compound xxvii (128.29 mg, 0.12 mmol, 1.0 equiv.) was added to dry tetrahydrofuran (1 mL) under inert gas protection, and a tetrahydrofuran solution containing 1 M tetrabutylammonium fluoride (0.15 ml, 0.30 mmol, 1.2 equiv.) was added under nitrogen protection at 0°C, and reacted at 0°C for 3 hours. After the reaction was terminated by TLC monitoring, the organic phase was extracted with dichloromethane (4×10 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. Compound xxviii (84.06 mg, yield 83.3%) was obtained by column chromatography purification (methanol: dichloromethane = 4:100) as a light yellow foam.

[0232] 1 H NMR (400 MHz, CDCl 3 )δ8.09-8.02(m,2H,Ar),8.00-7.94(m,2H,Ar),7.83-7.76(m,2H,Ar),7.74(d,J=2.0Hz,1H,Ar ),7.65(s,1H,Ar(DMNB)),7.64-7.57(m,1H,Ar),7.51-7.40(m,6H,Ar),7.35(t,J=7.8Hz,2H,Ar ),7.28-7.21(m,3H,Ar×2,Ar(DMNB)),6.12(dd,J=10.5,7.9Hz,1H,2-H),6.08(dd,J=3.4,1.1Hz ,1H,4-H),5.68(dd,J=10.5,3.4Hz,1H,3-H),5.49(d,J=7.9Hz,1H,1-H),4.98-4.83(m,2H,ArCH 2 ),4.66(s,2H,ArCH 2 ),4.42(td,J=6.3,1.2Hz,1H,5-H),3.91(d,J=1.7Hz,8H,ArOCH 3 ,6-H).

[0233] 13 C NMR (101 MHz, CDCl 3)δ165.7,165.6,165.2,154.0,148.4,147.7,141.3,139.0,137.2,137. 2,133.7,133.4,133.2,131.9,130.0,129.9,129.82,129.77,129.3,12 8.9,128.72,128.67,128.4,128.3,123.4,119.52,119.49,109.8,107. 9,100.9,73.3,71.7,70.3,69.1,68.7,68.0,63.39,63.37,56.6,56.3.

[0234] ESI-MS: found 861.2[M+Na] + .

[0235] (6) Synthesis of prodrug compound D:

[0236]

[0237] Specific operations:

[0238] (a) Under the protection of inert gas, compound xxviii (50 mg, 0.077 mmol, 1.0 equiv.) and Compretin (29 mg, 0.092 mmol, 1.2 equiv.) were dissolved in a dry tetrahydrofuran solution (1 mL), the mixture was cooled to 0°C in an ice bath and stirred for 15 minutes, and triphenylphosphine (22 mg, 0.0847 mmol, 1.1 equiv.) and diisopropyl azodicarboxylate (15 μL, 0.077 mmol, 1.0 equiv.) were added in sequence, and the reaction solution was heated from 0°C to room temperature and continued to react for 1 hour. The reaction solution was diluted with dichloromethane and washed twice with saturated ammonium chloride, saturated sodium bicarbonate, and saturated sodium chloride solutions in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by reduced pressure distillation using a rotary evaporator. The crude product was separated and purified by silica gel column chromatography (petroleum ether:ethyl acetate, 5:2, v / v) to give a light yellow oil xxix (51 mg, 0.054 mmol, yield 70%).

[0239] (b) Under the protection of inert gas, compound xxix (50 mg, 0.053 mmol, 1.0 equiv.) was dissolved in a mixture of methanol and dichloromethane (2:1, v / v) (1.5 mL), and a 5 mol / L sodium methoxide methanol solution (5 μL, 0.025 mmol, 0.5 equiv.) was added, and the mixture was stirred at room temperature for 12 hours. After the reaction of the raw material was completed as monitored by TLC, a hydrogen ion exchange resin was added to neutralize the reaction solution, the resin was filtered, and the resin was washed with a mixture of methanol and dichloromethane (2:1, v / v), and concentrated by reduced pressure distillation using a rotary evaporator. The crude product was separated and purified by silica gel column chromatography (dichloromethane: methanol, 9:1, v / v) to obtain compound D (32 mg, 0.039 mmol, yield 73%).

[0240] 1 H NMR (600 MHz, (CD 3 ) 2 SO)δ7.84(d,J=2.1Hz,1H,Ar),7.66(s,1H,Ar(DMNB)),7.48(dd,J=8.8,2.1Hz,1H,Ar),7 .41(d,J=8.8Hz,1H,Ar),7.22(s,1H,Ar(DMNB)),6.95-6.89(m,2H,Ar(CA4)),6.87(dd,J =8.4,1.9Hz,1H,Ar(CA4)),6.54(s,2H,Ar(CA4)),6.46(q,J=12.2Hz,2H,Ar(CA4)),5.23 (d,J=5.1Hz,1H,OH),5.13(d,J=7.7Hz,1H,1-H),5.02(s,1H,OH),4.92-4.80(m,4H,ArCH 2 ),4.77(d,J=4.5Hz,1H,OH),4.01(dd,J=7.5,4.4Hz,1H,5-H),3.83(s,3H,ArOCH 3 ),3.74-3.71(m,4H,4-H,ArOCH 3 ),3.71-3.67(m,4H,6-H,ArOCH 3 (CA4)),3.65-3.61(m,4H,6-H,ArOCH 3 (CA4)),3.61-3.57(m,7H,2-H,ArOCH 3 (CA4)),3.47(d,J=9.6Hz,1H,3-H).

[0241] 13 C NMR (151 MHz, (CD 3 )2 SO)δ153.9,153.1,149.6,149.1,147.8,147.5,140.3,139.6,137.3,133.5,132.9,131.2,130.4,129.8,129.7,129.2,124.3 ,122.8,117.3,115.2,112.6,110.3,108.6,106.5,101.3,74.4,73.7,70.4,69.4,69.1,69.0,60.5,56.5,56.4,56.14,56.11.

[0242] HRMS: m / z calculated [M+Na] + =847.2532, found 847.2532.

[0243] Example 5

[0244] HPLC verification that compounds A, B, C or D do not respond to β-galactosidase

[0245] The DMSO mother solution of compound A, B, C or D was diluted with 50% acetonitrile aqueous solution (v / v) to 100μM, 800μL solution. After fully dissolved, the 800μL solution was divided into two parts and the dissolved and diluted β-galactosidase (25U / mL) or an equal volume of water was added. After incubation in a 37°C shaker for 24 hours, the components in the solution were detected by HPLC. Mobile phase A was water and mobile phase B was acetonitrile. The gradient started from 50% B and changed to 90% B after 22 minutes. The HPLC ultraviolet detection wavelength was 254nm.

[0246] The experimental results are attached. Fig.10 , Compounds A, B, C or D did not respond to β-galactosidase when not responding to light.

[0247] Example 6

[0248] MTT assay of cytotoxicity of compound A, B, C or D on OVCAR3 cells:

[0249] OVCAR3 cells (3000 cells per well) or SKOV3 cells (2000 cells per well) were planted in the wells of a 96-well plate and incubated for 24 hours to allow the cells to grow stably. The culture medium was aspirated, and a DMEM culture medium solution containing 5% DMSO was added according to the concentration gradient. After incubation in a 37°C incubator containing 5% carbon dioxide for 3 hours, the illumination group was illuminated for 3 minutes, and other groups were not treated. For the double prodrug A culture medium, Cu (II) with a final concentration of 20 μM was also added. After continuing to incubate with the drug for 24 hours, the culture medium was washed out and the cells were washed 2-3 times with PBS buffer. After replacing the new culture medium, a DMEM solution containing 10% MTT solution (5 mg / mL PBS solution) was added to each well. The well plate was placed in a 37°C incubator containing 5% carbon dioxide for incubation for 3 hours. Finally, the culture medium was carefully aspirated and DMSO was added. After shaking on a shaker at 100 rpm for 10 minutes, the absorbance intensity at 490 nm of the well plate was detected, and the cell survival rate was calculated according to the formula:

[0250]

[0251] Among them, OD + OD is the absorbance value of the wells with MTT reagent, cells and drugs. control OD is the absorbance value of the wells with only MTT reagent added. - is the absorbance of the wells added with only MTT reagent and cells.

[0252] The experimental results are attached. Fig.11 , Fig.12 As shown in Table 1, compounds A, B, C and D can activate and controllably eliminate OVCAR3 or SKOV3 cells overexpressing β-galactosidase under light control, while their corresponding single prodrugs are highly toxic and uncontrollable to cells overexpressing β-galactosidase, indicating that compounds A, B, C and D have stronger controllability and lower toxic side effects than their corresponding single prodrugs. It can also be seen that compound A has stronger selectivity.

[0253] Table 1 IC of drugs against OVCAR3 cells 50 (nM)

[0254]

[0255] Example 7

[0256] Detection of the cell cycle inhibition of OVCAR3 cells by compound A:

[0257] 5 × 10 cells were seeded into each well of a 12-well plate. 5Cells were incubated for 24 hours to allow the cells to grow stably in the small dish. The culture medium was aspirated and a culture medium solution containing 0.5% DMSO was added according to the group. After a total incubation of 3 hours, the light group was illuminated for 3 minutes and incubated for another 24 hours. After the cell incubation, the cell culture medium was aspirated into a 1.5mL centrifuge tube, the adherent cells were washed once with PBS, and an appropriate amount of trypsin cell digestion solution was added to digest the cells. Incubate at room temperature until the adherent cells can be blown down by gentle blowing, and then the trypsin cell digestion solution was aspirated. Over-digestion of trypsin should be avoided during the process. Add the previously collected cell culture medium, gently blow down the cells, transfer them to a centrifuge tube, centrifuge at 1000g for 5 minutes, discard the supernatant, collect the cells, gently resuspend the cells with PBS and count them. Take 200,000 cells, centrifuge at 1000g for 5 minutes, and discard the supernatant. The collected cells were fixed with 75% ethanol at -20°C overnight, and the fixed cells were subsequently washed twice with PBS and stained with a staining solution containing RNaseA and propidium iodide for 15 minutes in the dark, and the cell cycle was analyzed by flow cytometry.

[0258] The experimental results are as follows Fig.13 As shown, it is demonstrated that compound A exerts its toxic effect by releasing CA4 drug molecules in terms of cell cycle inhibition, and has strong spatiotemporal controllability.

[0259] Example 8

[0260] Detection of the effect of compound A on apoptosis of OVCAR3 cells:

[0261] 5 × 10 cells were seeded into each well of a 12-well plate. 5Cells were incubated for 24 hours to allow the cells to grow stably in the small dish. The culture medium was aspirated and a medium solution containing 0.5% DMSO was added according to the group. After a total incubation of 3 hours, the light group was illuminated for 3 minutes and incubated for another 24 hours. After the cell incubation, the cell culture medium was aspirated into a 1.5mL centrifuge tube, the adherent cells were washed once with PBS, and an appropriate amount of trypsin cell digestion solution was added to digest the cells. Incubate at room temperature until the adherent cells can be blown down by gentle blowing, and then the trypsin cell digestion solution was aspirated. Over-digestion of trypsin should be avoided during the process. Add the previously collected cell culture medium, gently blow down the cells, transfer to a centrifuge tube, centrifuge at 1000g for 5 minutes, discard the supernatant, collect the cells, gently resuspend the cells with PBS and count them. Take 200,000 resuspended cells, centrifuge at 1000g for 5 minutes, discard the supernatant, add 195μL Annexin V-FITC binding solution and gently resuspend the cells. Add 5 μL Annexin V-FITC and 10 μL propidium iodide staining solution and mix gently. Incubate at room temperature (20-25°C) in the dark for 10-20 minutes, then place in an ice bath and use aluminum foil to protect from light. Resuspend the cells 2-3 times during the incubation process to improve the staining effect. Finally, use it for flow cytometry detection and immediately test it on the machine. Annexin V-FITC is green fluorescence and propidium iodide (PI) is red fluorescence. The effect of flow cytometry detection is shown.

[0262] The experimental results are as follows Fig.14 As shown, it was demonstrated that the cytotoxic effect of compound A in inducing cell apoptosis was dependent on ultraviolet light and had strong controllability.

[0263] Example 9

[0264] Compound A inhibits the scratch healing of OVCAR3 cells:

[0265] 5 × 10 cells were seeded into each well of a 6-well plate. 6 Cells were incubated for 36 hours to allow the cells to grow stably in the small dish, and then a scratch was made in the center of the well plate with a 200μL sterile micropipette tip, and the dropped cells were gently washed with PBS. According to the group, a medium (containing 2% FBS) solution containing 0.5% DMSO was added to the drug, and after incubation for 3 hours, the illumination group was illuminated with 365nm ultraviolet light for 3 minutes and continued to incubate for 48 hours. Finally, the cells were imaged with an optical microscope, and the wound healing rate was quantified using software.

[0266] The experimental results are as follows Fig.15 As shown, it is demonstrated that compound A can controllably release CA4 in inhibiting cell migration, and the cell migration inhibition effect depends on ultraviolet light and has strong controllability.

[0267] Example 10

[0268] According to the similar synthesis method of Examples 1-4, the cytotoxic drug molecules are replaced with other cytotoxic drugs shown in Table 2 to obtain the corresponding prodrug compounds E to Y.

[0269] Table 2

[0270]

[0271]

[0272] Compared with their corresponding single prodrugs, the double prodrug compounds E~Y also have strong spatiotemporal controllability and significantly reduce the risk of toxic side effects caused by their activation and release of drugs in non-tumor sites.

[0273] The embodiments provided above are not intended to limit the scope of the present invention, and the steps described are not intended to limit the execution order thereof. Those skilled in the art may make obvious improvements to the present invention in combination with existing common knowledge, which also fall within the scope of protection defined by the claims of the present invention.

Claims

1. A galactoside prodrug compound or a pharmaceutically acceptable salt thereof having the structure shown below: in, Z is a vacant bond or an ester group; R1 is derived from a cytotoxic drug containing a hydroxyl or amino group; R2 is selected from hydrogen atom, C 1-6 Alkyl, halogen, nitro, amino, carboxyl, C 1-6 Alkoxy, acylamino R a -CONH-, alkoxyacyl R b O-CO-, where R a , R b Independently selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 3-6 Cycloalkyl, halogenated C 3-6 Cycloalkyl, alkenyl, halogenated alkenyl, aromatic group; Any one of R3, R4, R5, and R6 is a photocage group that can break the ether bond and release the galactose hydroxyl group after light irradiation, and the other three are H; The photocage group is selected from R c Selected from hydrogen atoms or C 1-6 Alkyl; R d , R e independently selected from hydrogen, halogen, nitro, amino, C 1-6 alkylamino, substituted or unsubstituted five- to six-membered saturated heterocyclic group, substituted or unsubstituted phenyl, substituted or unsubstituted C 1-6 The substituted groups on the alkoxy, saturated heterocyclic group or phenyl group are selected from halogen, nitro, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylamino; C 1-6 The substituted groups on the alkoxy group are selected from halogen, nitro, C 1-6 Alkyl, halogenated C 1-6 alkyl.

2. The galactoside prodrug compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The galactoside prodrug compound is selected from the following structures:

3. The galactoside prodrug compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The galactoside prodrug compound is selected from the following compounds A, B, C, and D:

4. The galactoside prodrug compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The pharmaceutically acceptable salt includes inorganic salt or organic salt; the inorganic salt is selected from sodium salt, calcium salt, potassium salt, magnesium salt, silver salt, lithium salt; the organic salt is selected from meglumine salt, tromethamine salt, diethylamine salt, lysine salt, choline salt, arginine salt, tert-butylamine salt, N,N-dibenzylethylenediamine salt.

5. A solvate, enantiomer, diastereomer, tautomer, or a mixture thereof in any ratio of the galactoside prodrug compound according to any one of claims 1 to 3.

6. Use of the galactoside prodrug compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, or a solvate, enantiomer, diastereomer, tautomer or a mixture thereof in any proportion of the galactoside prodrug compound according to claim 5 in the preparation of an anti-tumor drug.

7. A composition, characterized in that A galactoside prodrug compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, or a solvent compound, enantiomer, diastereomer, tautomer or a mixture thereof in any proportion of the galactoside prodrug compound according to claim 5.

8. The composition according to claim 7, characterized in that The composition is a pharmaceutical composition, a dietary supplement, a food composition or a health product composition.

9. An anti-tumor drug, characterized in that: Containing the galactoside prodrug compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, or a solvent compound, enantiomer, diastereomer, tautomer or a mixture in any proportion of the galactoside prodrug compound according to claim 5, and a pharmaceutically acceptable carrier, excipient or diluent.

10. The anti-tumor drug according to claim 9, characterized in that: The pharmaceutically acceptable carrier is selected from microcapsules, microspheres, nanoparticles and liposomes; the drug preparation is selected from injections, freeze-dried powder injections, suspensions, implants, embolic agents, capsules, tablets, pills and oral solutions.

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