Synthesis method of anti-tumor and anti-angiogenesis drug PI-88 and precursor thereof

Abstract

The invention discloses a synthesis method of an anti-tumor and anti-angiogenesis medicine PI-88 and a precursor thereof, which comprises the following steps of: performing glycosylation coupling on a monosaccharide module and a double-chain fluorine carrier by adopting a polytetrafluoroethylene particle assisted purification strategy and a remote acyl and ortho-group participated high-stereoselectivity glycosylation strategy, and then removing a 2-bit protecting group to obtain a monosaccharide receptor; carrying out linear assembly to obtain mannose polysaccharide protected by a double-chain fluorine carrier, and then carrying out non-reducing end deprotection, phosphorylation-oxidation and selective protection group removal to obtain a PI-88 precursor; and carrying out sulfonation on exposed hydroxyl groups in the PI-88 precursor, and then removing other protecting groups to obtain PI-88. According to the method, the anti-tumor and anti-angiogenesis medicine PI-88 and the precursor thereof are rapidly and efficiently synthesized, the average yield of each step can reach 90% or above, only polytetrafluoroethylene particles are needed for purification in the assembly and purification process of target molecules, the product can be rapidly locked, the purification difficulty is reduced, the separation loss is reduced, and the method is suitable for industrial production. And large-scale synthesis is easy to realize.

Description

Technical Field

[0001] The invention belongs to the technical field of drug synthesis, and in particular relates to a method for synthesizing sulfated mannopentaose, sulfated mannotetraose and derivatives thereof, which are anti-tumor and anti-angiogenesis drugs. Background Art

[0002] Tumors are one of the main causes of death in my country and one of the main public health issues in my country. Therefore, people hope to find the genes related to the disease at the molecular level and then develop a series of prevention and treatment measures. With the continuous development of molecular biology, people have discovered the ten major mechanisms of tumor occurrence and development, among which angiogenesis is the key link in the tumor occurrence mechanism. In 1971, Professor Judah Folkman of Harvard Medical School first proposed the tumor angiogenesis hypothesis in the New Zealand Medical Journal, which mainly includes the following points: (1) The growth of early micro-tumors may not rely on the formation of new blood vessels. When the tumor diameter is larger than 1-2 mm, simple diffusion can no longer guarantee the oxygen and nutrient supply required for its growth; (2) When the tumor grows larger than 1-2 mm in diameter, tumor cells can induce the proliferation and migration of the surrounding vascular endothelial cells, forming new capillaries in the tumor mass, which can ensure the continued growth and metastasis of the tumor; (3) Tumor angiogenesis is induced by diffusible chemical signals generated by tumor cells, namely tumor angiogenesis factor (TAF); (4) The purpose of inhibiting tumor growth can be achieved by inhibiting the formation of tumor blood vessels; (5) Anti-tumor angiogenesis therapy does not directly kill tumor cells, but can inhibit the growth of tumor cells by blocking angiogenesis. This treatment method is called "dormancy induction". Therefore, inhibiting the angiogenesis process has become one of the recognized strategies for treating cancer. In terms of inhibiting tumor angiogenesis, VEGF / VEGFR is considered to be the most important anti-tumor therapeutic target. Common anti-angiogenic drugs currently on the market include large molecule single-target angiogenesis inhibitors such as bevacizumab, which is administered by intravenous injection. It was originally developed by Roche and launched in 2004. It is suitable for hepatocellular diseases, non-small cell lung diseases, etc. It is an effective anti-angiogenic treatment plan. It is a type of VEGF monoclonal antibody that targets vascular endothelial growth factor (VEGF) and competitively inhibits the binding of vascular endothelial cell surface receptors to VEGF, thereby exerting an anti-angiogenic effect. Another common anti-angiogenic drug is a small molecule multi-target angiogenesis inhibitor such as sorafenib tosylate, which is an oral drug developed by Bayer Pharmaceuticals in Germany. It acts on tumor blood vessels and tumor cells at the same time, has a dual anti-tumor effect, and can be used to treat hepatocellular carcinoma that cannot be operated on or has distant metastases. Although these drugs have obvious clinical anti-tumor effects, they still have problems such as low survival rate and toxic side effects on the heart. Therefore, the development of angiogenesis inhibitors with low side effects and the ability to inhibit multiple angiogenic factors is the current research direction.

[0003] With the continuous development of sugar chemistry, sugar drugs and sugar vaccines have been used to manufacture and prevent various diseases. The heparin sulfate mimetic PI-88 developed by Vito Ferro's group at Progen in Australia is a complex mixture of highly sulfonated mannooligosaccharides with anti-angiogenesis and anti-tumor metastasis properties. Currently, it is undergoing Phase III clinical trials for the treatment of post-resection hepatocellular carcinoma. As a heparin sulfate mimetic, PI-88 can simultaneously inhibit angiogenesis and tumor metastasis through the following two mechanisms: (1) Direct inhibition of angiogenesis: interfering with the interaction between HS and angiogenic growth factors FGF-1, FGF-2, and VEGF, thereby avoiding the cell signaling cascade that triggers angiogenesis. (2) Blocking tumor metastasis: By strongly inhibiting heparinase, it prevents it from degrading the proteoglycan HS side chains on the extracellular matrix and basement membrane, thereby preventing metastatic tumor cells from escaping into the blood circulation. PI-88 is derived from the extracellular phosphomannan of Pichia pastoris. Its main components are obtained by mild acid-catalyzed hydrolysis. It is mainly pentasaccharide, with very few tetrasaccharides, extremely few disaccharides and hexasaccharides. It is currently difficult to separate them by gel exclusion chromatography. Therefore, obtaining the most active and single-component sulfonated mannotetraose and sulfonated mannopentaose by chemical synthesis is conducive to the development of new single-component tumor anti-angiogenic medicinal molecules, which are expected to be used in future tumor treatment.

[0004] For the sulfonated mannotetraose and sulfonated mannopentose, the main sugar link is α-(1-2)- / α-(1-3)-linked manno-oligosaccharide. In the prior art, the construction method is mainly synthesized by glycosylation reaction with glucosinolate or imidate as donor. For the glycosylation reaction of glucosinolate, glucosinolate has good stability and moderate activity, and NIS is often used as a catalyst. However, the use of glucosinolate synthesis is less, which may be due to the inherent low activity of mannose donors and the inherent low activity of glucosinolate donors. These two factors together lead to the inability of mannose glucosinolate donors to be effectively activated under glycosylation conditions. For imidate donors, the donors are unstable and not suitable for storage. Therefore, they are not suitable for large-scale preparation, but they have high activity and good reaction. During the reaction, the catalyst TMSOTf is pre-mixed with the acceptor, and then the trichloroacetimidate acceptor is added for glycosylation reaction. For the synthesis of non-oligosaccharide glycosides, a very high yield can be obtained. Therefore, rational design of the protecting group of the hydroxyl group in the mannose imidate donor is the main way to regulate its reactivity and stereoselectivity, and is also an important method to realize the application of glycosylation reaction with mannose glucosinolate as donor in the preparation of α-(1-2)- / α-(1-3)-linked manno-oligosaccharide chains.

[0005] In the separation and purification of glycosylation products, the traditional method uses column chromatography technology, which has certain disadvantages, such as long separation time, product loss, limited separation capacity, and limited column life, which add some obstacles to the synthesis process. Chai Yonghai's research group first proposed the strategy and idea of ​​"liquid phase reaction, solid phase separation", and applied the synthesis method of oligosaccharides assisted by polytetrafluoroethylene particles, which can obtain purified products without column chromatography. The advantages of this method are: (1) Simple and efficient: liquid phase reaction, easy to detect the reaction progress, but no large excess of sugar modules are required; solid phase separation, the compound can be purified by simple filtration operation; (2) Rapid locking of target products: due to the hydrophobic characteristics of fluorinated compounds, the target compound can be quickly locked from complex reactants; (3) Easy removal of fluorine carriers: the research group has developed two types of benzyl fluorine carriers, one is a conventional benzyl fluorine carrier, which can be removed by hydrogenolysis at the same time as the benzyl protecting group on the sugar ring; the other is a photosensitive benzyl fluorine carrier, which can be efficiently removed by photolysis without affecting the conventional benzyl protecting group on the sugar ring; these two types of fluorine carriers can meet the linear or convergent synthesis requirements of different types of sugar synthesis; (4) High carrying capacity: So far, the number of sugar units supported by the fluorine carrier developed by the research group can reach 18 saccharides. Although the upper limit is still to be studied, the current results have fully proved that the existing fluorine carriers can be widely used in the synthesis of oligosaccharides and even polysaccharides. In addition, the research group has recently optimized the synthesis of fluorine carriers, simplified the synthesis steps and improved the reaction yield. Summary of the invention

[0006] The purpose of the present invention is to provide a method for highly selectively synthesizing an anti-tumor and anti-angiogenic drug PI-88 and its precursor, wherein the method couples a monosaccharide module with a double-chain fluorine carrier by glycosylation, removes the 2-position protecting group to obtain a monosaccharide receptor, and then linearly assembles to obtain a double-chain fluorine carrier-protected mannopolysaccharide, and then undergoes non-reducing end deprotection, phosphitylation-oxidation, and selectively removes the protecting group to obtain a PI-88 precursor. The exposed hydroxyl group in the PI-88 precursor is then sulfonated and the remaining protecting groups are removed to obtain the anti-tumor and anti-angiogenic drug PI-88. The assembly and purification process of the target molecule only requires purification by polytetrafluoroethylene particles, which can not only quickly lock the product, but also reduce the purification difficulty and separation loss.

[0007] The method of the anti-tumor and anti-angiogenesis drug PI-88 and its precursor provided by the present invention comprises the following steps:

[0008] Step 1: Synthesis of monosaccharide modules

[0009] Using D-mannose as a raw material, compound I is obtained by protecting its hydroxyl position, and after removing the R1 protecting group at position 1 of compound I, it is reacted with trichloroacetonitrile to obtain a monosaccharide module A, as shown in route (1):

[0010]

[0011] Using D-mannose as a raw material, compound II is obtained by protecting its hydroxyl position, and after removing the R1 protecting group at position 1 of compound II, it is reacted with trichloroacetonitrile to obtain a monosaccharide module B, as shown in route (2):

[0012]

[0013] Using D-mannose as a raw material, compound II is obtained by protecting its hydroxyl position, and after removing the R1 protecting group at position 1 of compound II, it is reacted with trichloroacetonitrile to obtain monosaccharide module C, as shown in route (3):

[0014]

[0015] Among them, the R1 protecting group is selected from any one of acetyl (Ac), ethylthio (EtS), p-methylphenylthio (STol), o-iodobenzylthio (SOIB), and benzylthio (PhCH2S); the R2, R3, R4, and R5 protecting groups are each independently selected from any one of acetyl (Ac), benzoyl (Bz), benzyl (Bn), p-methoxybenzyl (PMB), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBS), and trityl (Tr); the R6 protecting group is selected from any one of ester protecting groups such as acetyl and levulinyl (Lev), and the R6 protecting group is different from the R1, R3, R4, and R5 protecting groups.

[0016] Step 2: Synthesis of double-stranded fluorophore-protected mannopolysaccharide

[0017] The double-chain fluorine carrier D and the monosaccharide module C are glycosylated to obtain monosaccharide E, and then the 2-position R6 protecting group is removed to obtain monosaccharide receptor F; the monosaccharide receptor F is repeatedly glycosylated with the monosaccharide module A for n times and then the 3-position Lev protecting group is removed to obtain compound G; compound G is then glycosylated with the monosaccharide module B to obtain double-chain fluorine carrier protected mannopolysaccharide H, as shown in route (4):

[0018]

[0019] Here, n is an integer of 1-3, a is an integer of 0-2, and b is an integer of 4-7.

[0020] Step 3: Synthesis of PI-88 and its precursors

[0021] The Lev protecting group at the 6-position of the terminal sugar unit in the double-stranded fluorine carrier-protected mannopolysaccharide H is removed and phosphorylated, and the remaining protecting groups are selectively removed according to the structure of the target compound to obtain PI-88 precursors with different numbers of exposed hydroxyl groups; the exposed hydroxyl groups in the PI-88 precursor are further sulfonated, and then all the remaining protecting groups are removed to obtain PI-88.

[0022] In the above step 1, it is preferred that the R1 protecting group is p-methylphenylthio, and the R2, R3, R4, and R5 protecting groups are each independently selected from any one of benzoyl, benzyl, p-methoxybenzyl, tert-butyldiphenylsilyl, tert-butyldimethylsilyl, and trityl; and the R6 protecting group is acetyl.

[0023] Furthermore, in the above step 2, the method for glycosylation coupling of the double-chain fluoro carrier D and the monosaccharide module C is: the double-chain fluoro carrier D and the monosaccharide module C are mixed and dissolved in a solvent, an acid catalyst is added, and the reaction is carried out at a temperature of -20 to -40°C for 0.5 to 2h; the solvent is selected from any one of dichloromethane, tetrahydrofuran, toluene, and acetonitrile, or a mixture of two of them; the acid catalyst is trimethylsilyl trifluoromethanesulfonate or trifluoromethanesulfonic acid.

[0024] Furthermore, in the above step 2, the method for glycosylation coupling of the monosaccharide receptor F and the monosaccharide module A is: the monosaccharide receptor F and the monosaccharide module A are mixed and dissolved in a solvent, an acid catalyst is added, and the reaction is carried out at a temperature of -20 to -40°C for 0.5 to 2h; the solvent is selected from any one of dichloromethane, tetrahydrofuran, toluene, and acetonitrile, or a mixture of two thereof; the acid catalyst is trimethylsilyl trifluoromethanesulfonate or trifluoromethanesulfonic acid.

[0025] Furthermore, in the above step 2, the method for removing the 3-position Lev protecting group is: using dichloromethane or a pyridine / acetic acid mixed solution as a solvent, adding a methanol solution of hydrazine acetate, and reacting at room temperature for 2 to 6 hours.

[0026] Furthermore, in the above step 2, the method for glycosylation reaction of the compound G and the monosaccharide module B is: mixing the compound G and the monosaccharide module B and dissolving them in a solvent, adding an acid catalyst, and reacting at a temperature of -20 to -40°C for 0.5 to 2h; the solvent is selected from any one of dichloromethane, tetrahydrofuran, toluene, and acetonitrile or a mixture of two thereof; the acid catalyst is trimethylsilyl trifluoromethanesulfonate or trifluoromethanesulfonic acid.

[0027] Furthermore, in the above step 3, the method for removing the Lev protecting group at the 6-position of the terminal sugar unit in the double-stranded fluorine carrier protected mannopolysaccharide H is: dissolving mannopolysaccharide H in dichloromethane or a pyridine / acetic acid mixed solution, then adding a methanol solution of hydrazine acetate, and reacting at room temperature for 2 to 6 hours.

[0028] Furthermore, in the above step 3, the phosphorylation method is: using dichloromethane as a solvent, adding tetrazole and dibenzyl-N,N-diisopropylphosphoramidite, reacting at room temperature for 0.5 to 2 hours, cooling to -20 to -40°C, adding m-chloroperbenzoic acid and reacting for another 0.5 to 2 hours.

[0029] Furthermore, in the above step 3, the sulfonation method is: mixing the PI-88 precursor and the sulfur trioxide salt complex, dissolving the mixture in a solvent, and subjecting the mixture to a microwave reaction at 100 to 140° C. for 0.3 to 1 h; or mixing the PI-88 precursor and the sulfur trioxide salt complex, dissolving the mixture in a solvent, and subjecting the mixture to a reaction at 50 to 80° C. for 1 to 3 d; the solvent is selected from any one or two of triethylamine, tetrahydrofuran, pyridine, and N,N-dimethylformamide; and the sulfur trioxide salt complex is selected from any one of sulfur trioxide trimethylamine salt complex, sulfur trioxide triethylamine salt complex, and sulfur trioxide pyridine salt complex.

[0030] The beneficial effects of the present invention are as follows:

[0031] The present invention first designs and synthesizes monosaccharide modules A, B and C, connects the monosaccharide module C with a double-chain fluorine carrier, deprotects the monosaccharide module C to obtain a monosaccharide receptor, and then linearly assembles the monosaccharide module C with the monosaccharide modules A and B to obtain mannopolysaccharide protected by a double-chain fluorine carrier, and then undergoes non-reducing end deprotection, phosphitylation-oxidation, and selectively removes the protecting groups to obtain a PI-88 precursor; then, the exposed hydroxyl group in the PI-88 precursor is sulfonated and the remaining protecting groups are removed to obtain the anti-tumor and anti-angiogenesis drug PI-88. The significant advantages of the preparation method of the present invention are: (1) the introduction of fluorine-containing compounds can not only quickly lock the target product, but also make the product easy to separate and purify, and the fluorine compound can be recycled; (2) the protecting group of the hydroxyl group in the donor and acceptor is reasonably designed, and a high stereoselective glycosylation strategy involving remote acyl groups and neighboring groups is adopted to achieve glycosylation to construct α-(1-2)- / α-(1-3)-linked manno-oligosaccharide chains, and finally achieve rapid and efficient preparation of the anti-tumor and anti-angiogenic drug PI-88 and its precursors; (3) by designing the hydroxyl protecting groups of the monosaccharide modules A, B, and C, a PI-88 precursor with exposed hydroxyl groups at different sites is obtained, which is helpful to obtain the anti-tumor and anti-angiogenic drug PI-88 with a determined sulfonation site, and lays a foundation for its research in the biomedical field. DETAILED DESCRIPTION

[0032] The present invention is further described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited to these embodiments.

[0033] Example 1

[0034] Step 1: Synthesis of sugar modules

[0035] (1) D-mannose (10 g, 55.5 mmol), sodium acetate (NaOAc, 2.27 g, 27.75 mmol) and acetic anhydride (AC2O, 52.5 mL, 555 mmol) were added to a 100 mL reaction bottle. After reflux reaction at 140°C for 3 h, the reaction was completed after monitoring by TLC plate. The mixture was washed with saturated NaHCO3 aqueous solution until neutral, extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate and filtered, and concentrated in vacuo to obtain compound 1.

[0036]

[0037] (2) To compound 1 (21.66 g, 55.5 mmol), p-methylthiophenol (TolSH, 10.34 g, 83.25 mmol) was added, dissolved in dry dichloromethane (150 mL), and then boron trifluoride etherate (BF3·Et2O, 21.0 mL, 166.5 mmol) was added dropwise. After reacting at room temperature for 3 h, the reaction was monitored by TLC plate. The mixture was washed with saturated NaHCO3 aqueous solution until neutral, extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate and filtered, concentrated in vacuo, and separated by column chromatography (ethyl acetate / petroleum ether=1 / 2, v / v) to obtain compound 2 (23.21 g, 51.06 mmol, yield 92%), the structural characterization data of which are as follows: 1 H NMR (400MHz, CDCl3) δ7.35(d,J=8.0Hz,2H),7.10(d,J=8.0Hz,2H),5.47(s,1H),5.40(s,1H),5.30( d,J=6.1Hz,2H),4.54(s,1H),4.27(s,1H),4.08(d,J=14.2Hz,1H),2.30(s,3H),2.08–1.97(m,12H); 13 C NMR (100MHz, CDCl3) δ170.01,169.91,138.54,130.05,129.93,128.85,86.08,70.93,69.48,69.44,66.47,62.57,21.2,20.78; HRMS (ESI) m / z C 21 H 26 NaO9S(M+Na) + Theoretical value 477.1190, measured value 477.1194.

[0038]

[0039] (3) Compound 2 (23.21 g, 51.06 mmol) was dissolved in dry methanol (MeOH, 102.1 mL), and freshly prepared sodium methoxide (MeONa, 5.106 mmol Na dissolved in 10 mL dry MeOH) was added. The mixture was reacted at room temperature for 3 h. The complete reaction was monitored by TLC plate. Amberlite (R) IRC 120H resin was added to adjust the pH to 7, and the mixture was concentrated in vacuo to obtain compound 3.

[0040]

[0041] (4) Compound 3 (4.4417 g, 15.51 mmol) was dissolved in dry pyridine (pyridine, 51.7 mL), tert-butyldiphenylsilyl chloride (TBDPSCl, 3.6 mL, 20.32 mmol) was added, and the mixture was reacted at room temperature for 12 h. The reaction was monitored by TLC plate, and the mixture was concentrated in vacuo. 1 mol / L HCl aqueous solution and saturated NaHCO3 aqueous solution were added to adjust the pH to neutral, and the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and then filtered, concentrated in vacuo, and separated by column chromatography (ethyl acetate / petroleum ether = 1 / 1, v / v) to obtain compound 4 (7.65 g, 14.58 mmol, yield 94%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ7.69(d,J=7.9Hz,4H),7.41–7.35(m,8H),7.03(d,J=8.1Hz,2H),4.77(s,1H),4.12(s,1H),3. 92(s,2H),3.79(t,J=9.3Hz,1H),3.54(d,J=8.1Hz,1H),3.44–3.34(m,3H),3.02(s,1H),2.30(s,3H),1.05(s,9H); 13 C NMR (100MHz, CDCl3) δ137.54,135.58,132.83,132.73,131.59,129.83,129.74,127 .79,87.45,79.21,75.04,72.04,69.55,64.83,26.77,21.07,19.15; HRMS(ESI)m / z C 29 H 36 NaO5SSi(M+Na) + Theoretical value is 547.1945, measured value is 547.1944.

[0042]

[0043] (5) Compound 4 (7.65 g, 14.58 mmol) was mixed with dibutyltin oxide (Bu2Sn, 3.9927 g, 16.04 mmol), dissolved in dry toluene (97.2 mL), and refluxed at 140°C for 6 h. Tetrabutylammonium bromide (TBAB, 5.2288 g, 16.22 mmol) and 4-methoxybenzyl chloride (PMBCl, 3.1 mL, 22.75 mmol) were then added and reacted at 50°C for 30 h. The reaction was monitored by TLC plate. After vacuum concentration, the mixture was directly subjected to column chromatography (ethyl acetate / petroleum ether = 1 / 2, v / v) to obtain compound 5 (9.029 g, 14.00 mmol, yield 96%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ7.68(dd,J=7.5,4.0Hz,4H),7.43–7.28(m,10H),7.02(d,J=7.8Hz,2H),6.92(d,J=8.4Hz,2H),5.47(s,1H),4.6 7(s,2H),4.20(d,J=4.1Hz,2H),4.02(t,J=9.2Hz,1H),3.92–3.88(m,2H),3.82(s,3H),3.73–3.68(m,1H),2.29(s,3H),1.05(s,9H); 13 C NMR (100MHz, CDCl3) δ159.20,137.07,135.41,135.30,133.00,132.71,131.76,130.09,129.41,127.4 4,87.74,87.69,79.01,72.64,71.37,69.06,64.33,54.86,26.55,25.89,20.80,18.94; HRMS(ESI)m / zC 37 H 44 NaO6SSi(M+Na) + Theoretical value: 667.2520, measured value: 667.2515.

[0044]

[0045] (6) Compound 5 (9.029 g, 14.00 mol) was dissolved in dry N,N-dimethylformamide (DMF, 70 mL), and NaH (1.96 g, 49 mmol, purity 60%) was quickly added under ice bath. After 10 min, benzyl bromide (BnBr, 5.8 mL) was added dropwise. After 3 h of reaction, the reaction was complete when monitored by TLC plate. The reaction solution was quenched with ethanol under ice bath cooling, diluted with ethyl acetate, poured into water and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and filtered, concentrated in vacuo, and separated by column chromatography (ethyl acetate / petroleum ether = 1 / 4, v / v) to obtain compound 6 (11.32 g, 13.72 mmol, yield 98%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ7.81–7.69(m,4H),7.41(d,J=9.0Hz,5H),7.36(d,J=2.8Hz ,5H),7.36–7.30(m,5H),7.31(s,10H),7.07(dd,J=7.8,4.8Hz,2H),6.94–6.86(m ,2H),5.58(s,1H),5.04–4.92(m,1H),4.20(d,J=5.5Hz,1H),4.09–4.04(m,1H), 4.01(t,J=4.8Hz,1H),3.96–3.88(m,2H),3.84(s,3H),2.34(s,3H),1.08(s,9H); 13 C NMR (100MHz, CDCl3) δ158.97,137.90,135.25,131.31,131.01,129.99,129.41,129.19,128.63,127.97,127.35,113. 48,85.77,79.73,77.00,76.68,74.90,74.41,73.76,71.66,71.44,62.81,54.63,33.10,26.48,18.97; HRMS(ESI)m / z C 51 H 56 NaO6SSi(M+Na) + Theoretical value is 847.3459, measured value is 847.3467.

[0046]

[0047] (7) Compound 6 (11.32 g, 13.72 mmol) was dissolved in dichloromethane (91.5 mL) and water (5.1 mL), and 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ, 4.67 g, 20.58 mmol) was added. After reacting at room temperature for 6 h, the reaction was completed after monitoring by TLC plate. Saturated aqueous NaHCO3 solution was added to neutralize the mixture, and the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and then filtered, concentrated in vacuo, and separated by column chromatography (ethyl acetate / petroleum ether = 1 / 4, v / v) to obtain compound 7 (7.83 g, 11.11 mmol, yield 94%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ8.04(d,J=7.4Hz,2H),7.99(d,J=7.2Hz,2H),7.56(ddt,J=30.0,13.8,7.6Hz,20H),5.91(s,1H),5.22(d,J =11.1Hz,1H),4.94(d,J=5.5Hz,1H),4.91(d,J=4.8Hz,1H),4.79(d,J=11.8Hz,1H),4.37–4.16(m,6H),2.52(s,3H),1.37(s,9H); 13 C NMR (100MHz, CDCl3) δ138.09,137.27,136.72,135.50,135.13,133.35,132.69,131.17,130.67,129.31,128.0 4,127.92,127.41,84.87,79.79,76.48,76.15,72.84,71.94,71.68,62.58,26.39,20.67,18.89; HRMS(ESI)m / z C 43 H 48 NaO5SSi(M+Na) + Theoretical value: 727.2884, measured value: 727.2891.

[0048]

[0049] (8) Compound 7 (7.83 g, 11.11 mmol) was dissolved in anhydrous dichloromethane (37 mL), and N,N'-diisopropylcarbodiimide (DIPC, 2.6 mL, 16.67 mmol), 4-dimethylaminopyridine (DMAP, 1.26 g, 11.11 mmol), and levulinic acid (LevOH, 1.7 mL, 16.67 mmol) were added in sequence. The mixture was stirred at room temperature for 6 h. After vacuum concentration, the mixture was directly purified by column chromatography (ethyl acetate / petroleum ether = 1 / 4, v / v) to obtain compound 8 (6.77 g, 8.44 mmol, yield 76%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ7.49–7.28(m,20H),7.00(d,J=8.5Hz,4H),5.61(s,1H),5.33(dd,J=8.7,3.2Hz,1H),4.84(d,J=11.2Hz,1H),4.72(q,J=12.1Hz,3 H),4.34(d,J=9.1Hz,2H),4.23(s,1H),4.11(dd,J=11.4,3.2Hz,1H),3.97( d,J=11.1Hz,1H),2.75–2.53(m,4H),2.34(s,3H),2.17(s,3H),1.13(s,9H); 13 C NMR (100MHz, CDCl3) δ205.99,171.85,138.04,137.80,136.98,135.71,13 5.32,133.55,132.81,131.68,131.37,130.76,129.66,129.48,129.32,1 28.13,128.10,127.50,127.45,127.40,77.50,74.61,74.09,73.43,73.0 8,72.19,62.67,37.50,29.52,27.80,26.53,20.84,19.08; HRMS(ESI)m / z C 48 H 54 NaO7SSi(M+Na) + Theoretical value is 825.3252, measured value is 825.3258.

[0050]

[0051] (9) Compound 8 (6.77 g, 8.44 mmol) was dissolved in dry tetrahydrofuran (THF, 42.2 mL) and dry pyridine (42.2 mL), and hydrofluoric acid pyridinium salt complex (HF / py, 18.4 mL, 185.7 mmol) was added dropwise under ice bath. After reacting for 10 h, the reaction was completed after monitoring by TLC plate. Saturated NaHCO3 aqueous solution was added to adjust the mixture to neutrality, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and filtered, concentrated in vacuo, and separated by column chromatography (ethyl acetate / petroleum ether = 1 / 3, v / v) to obtain compound 9 (4.0 g, 7.09 mmol, yield 84%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ7.36 (q, J=6.6, 6.1Hz, 12H), 7.13 (d, J=7.9Hz, 2H), 5.45 (s ,1H),5.29(dd,J=9.2,3.1Hz,1H),4.79(d,J=11.4Hz,1H),4.72(s,1H),4.66(s,1 H),4.60(d,J=12.0Hz,1H),4.23(d,J=9.6Hz,1H),4.14(d,J=9.5Hz,2H),3.84(s ,2H),2.68(dt,J=12.0,6.4Hz,2H),2.56–2.43(m,2H),2.35(s,3H),2.16(s,3H); 13 C NMR (100MHz, CDCl3) δ206.12,171.86,137.95,137.73,137.49,132.30,129.70,127.75,127.67,12 7.60,85.91,74.61,73.90,72.93,72.84,72.52,61.58,37.52,29.59,27.74,20.92; HRMS(ESI)m / z C 32 H 36 NaO7S(M+Na) + Theoretical value is 587.2074, measured value is 587.2085.

[0052]

[0053] (10) Compound 9 (4.0 g, 7.09 mmol) was dissolved in dry pyridine (23.6 mL), and benzoyl chloride (BzCl, 1.8 mL, 15.6 mmol) was added dropwise under ice bath. After reacting for 3 h, the mixture was concentrated in vacuo. The concentrated product was dissolved in dichloromethane. The reaction was monitored by TLC plate. 1 mol / L HCl aqueous solution and saturated NaHCO3 aqueous solution were added to neutralize the mixture, and the mixture was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and then filtered, concentrated in vacuo, and separated by column chromatography (ethyl acetate / petroleum ether = 1 / 2, v / v) to obtain compound I-1 (4.68 g, 7.02 mmol, yield 99%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ8.06(d,J=7.8Hz,2H),7.48–7.25(m,15H),7.05(d,J=7.8Hz,2H),5.60(d,J=1.8Hz,1H),5.38(dd,J=9. 5,3.2Hz,1H),4.84(d,J=11.2Hz,1H),4.79–4.58(m,6H),4.25(d,J=8.8Hz,2H),2.77–2.52(m,4H),2.31(s,3H),2.18(s,3H); 13 C NMR (100MHz, CDCl3) δ205.93,171.77,165.93,137.57,137.45,132.72,131.88,129.72,129.57,129.48,128.20,128.1 5,128.06,127.65,127.47,85.37,74.56,74.05,73.10,72.18,70.48,63,47,37.44,29.49,27.72,20.83; HRMS(ESI)m / z C 39 H 40 NaO8S(M+Na) + Theoretical value is 691.2336, measured value is 691.2343.

[0054]

[0055] (11) Compound I-1 (4.68 g, 7.02 mmol) was dissolved in a mixed solution of acetone (70 mL) and H2O (3.7 mL), and N-bromosuccinimide (NBS, 5.0 g, 28.1 mmol) was added under ice bath. After reacting for 3 h, the reaction was monitored by TLC plate and concentrated in vacuo. Saturated aqueous Na2S2O3 solution was added to the concentrated product, and the product was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and filtered, and then azeotroped with toluene to remove water. The product was dissolved in anhydrous dichloromethane (15.3 mL), and Cs2CO3 (686.1 mg, 2.1 mmol) was added. Redistilled trichloroacetonitrile (CNCCl3, 2.5 mL, 25.3 mmol) was added dropwise. After reacting at room temperature for 3 h, the product was filtered and concentrated in vacuo to obtain monosaccharide module A-1.

[0056]

[0057] (12) Compound 7 (1.05 g, 1.5 mmol) was dissolved in dry pyridine (5 mL), and benzoyl chloride (0.4 mL, 1.8 mmol) was added dropwise under ice bath. After reacting for 3 h, the reaction was completed after monitoring by TLC plate, and the product was concentrated in vacuo. The concentrated product was dissolved in dichloromethane, and 1 mol / L HCl aqueous solution and saturated NaHCO3 aqueous solution were added to neutralize the product. The product was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and filtered. After vacuum concentration, the product was separated by column chromatography (ethyl acetate / petroleum ether = 1 / 3, v / v) to obtain compound 10 (1.13 g, 1.39 mmol, yield 93%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ8.07 (d, J=7.8Hz, 2H), 7.72 (dd, J=15.5, 7.3Hz, 4H), 7.48–7.2 9(m,16H),7.17(s,3H),7.06(d,J=7.7Hz,4H),5.59(s,1H),5.49(dd,J=9.3,3.4Hz,1 H),4.77(d,J=10.7Hz,1H),4.73–4.63(m,2H),4.55(d,J=12.5Hz,2H),4.39(t,J=9. 5Hz,1H),4.34–4.22(m,2H),4.06(dd,J=11.2,4.0Hz,1H),2.33(s,3H),1.06(s,9H); 13C NMR (100MHz, CDCl3) δ165.67,137.56,134.73,133.09,130.49,129.72,128.23,127.81,127.62 ,127.43,85.79,77.63,73.41,73.35,72.41,72.18,62.90,26.71,26.49,21.05; HRMS(ESI)m / z C 50 H 52 NaO6SSi(M+Na) + Theoretical value is 831.3146, measured value is 831.3142.

[0058]

[0059] (13) Compound 10 (1.13 g, 1.39 mmol) was dissolved in dry tetrahydrofuran (13.9 mL) and dry pyridine (13.9 mL), and hydrofluoric acid pyridinium salt complex (2.8 mL, 30.6 mmol) was added dropwise under ice bath. After reacting for 10 h, the reaction was completed after monitoring by TLC plate. Saturated NaHCO3 aqueous solution was added to adjust the mixture to neutrality, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and filtered, concentrated in vacuo, and separated by column chromatography (ethyl acetate / petroleum ether = 1 / 3, v / v) to obtain compound 11 (634.1 g, 1.11 mmol, yield 84%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ8.12(d,J=7.7Hz,2H),7.64(t,J=7.5Hz,1H),7.51(t,J=7.7Hz,3H),7.4 3(d,J=7.9Hz,3H),7.28(s,5H),7.18(t,J=8.3Hz,5H),5.60(dd,J=8.9,3.4Hz,1H),5.57(d,J =1.8Hz,1H),4.86(d,J=11.0Hz,1H),4.76(d,J=11.0Hz,1H),4.69(d,J=12.2Hz,1H),4.56(d, J=12.1Hz,1H),4.37(d,J=7.9Hz,2H),4.36–4.29(m,1H),3.93(d,J=2.7Hz,2H),2.38(s,3H); 13C NMR (100MHz, CDCl3) δ165.80,138.15,137.97,133.44,132.73,130.12,129.95,128.68,128. 58,128.53,128.17,128.00,86.24,75.24,74.64,73.39,72.73,62.02,21.32; HRMS(ESI)m / z C 34 H 34 NaO6S(M+Na) + Theoretical value 593.1968, measured value 593.1968.

[0060]

[0061] (14) Compound 11 (634.1 g, 1.11 mmol) was dissolved in anhydrous dichloromethane (7.4 mL), and N,N'-diisopropylcarbodiimide (0.3 mL, 1.67 mmol), 4-dimethylaminopyridine (135.6 mg, 1.11 mmol), and levulinic acid (0.2 mL, 1.67 mmol) were added in sequence. The mixture was stirred at room temperature for 6 h. The reaction was monitored by TLC plate. After vacuum concentration, the mixture was directly purified by column chromatography (ethyl acetate / petroleum ether = 1 / 4, v / v) to obtain compound II-1 (623.1 mg, 0.93 mmol, yield 84%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ8.08(d,J=7.7Hz,2H),7.61(t,J=7.4Hz,1H),7.48(t,J=7.7Hz,3H),7.39(d,J=7.9Hz,3 H),7.24–7.21(m,5H),7.17–7.12(m,5H),5.55(s,1H),5.50(dd,J=9.3,3.0Hz,1H),4.78(d,J=10.7Hz,1H),4 .67(d,J=12.0Hz,1H),4.62(d,J=10.8Hz,1H),4.50(d,J=12.0Hz,1H),4.48–4.38(m,2H),4.33(d,J=10.7Hz, 1H),4.25(s,1H),4.18(t,J=9.4Hz,1H),2.77–2.68(m,2H),2.62(d,J=6.6Hz,2H),2.34(s,3H),2.16(s,3H); 13C NMR (100MHz, CDCl3) δ206.37,172.34,165.41,137.76,137.38,137.26,133.20,132.16,129.75,129.65,128.40,128.28,128.2 1,127.94,127.77,127.67,85.57,76.93,74.91,74.38,74.33,72.21,70.52,63.30,37.72,29.69,27.75,21.01; HRMS(ESI)m / zC 39 H 40 NaO8S(M+Na) + Theoretical value is 691.2336, measured value is 691.2343.

[0062]

[0063] (15) Compound II-1 (623.1 mg, 0.93 mmol) was dissolved in a mixed solution of acetone (9.3 mL) and water (0.5 mL), and N-bromosuccinimide (662.1 mg, 3.72 mmol) was added under ice bath. After reacting for 3 h, the reaction was completed after monitoring by TLC plate. The acetone was removed by vacuum concentration, and a saturated aqueous Na2S2O3 solution was added to the concentrated product, which was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and filtered, and then separated by column chromatography (ethyl acetate / petroleum ether = 1 / 1, v / v) to obtain compound 12 (517.6 mg, 0.92 mmol, yield 99%). The structural characterization data are as follows: 1 H NMR (400MHz, CDCl3) δ8.08(d,J=7.5Hz,2H),7.60(t,J=7.3Hz,2H),7.47(t,J=7.7Hz,2H),7.29–7.27(m,2 H),7.23–7.16(m,8H),5.63(dd,J=9.2,3.2Hz,1H),5.31(s,1H),4.76(d,J=10.8Hz,1H),4.65(d,J=12.0H z,1H),4.60(t,J=11.4Hz,2H),4.43(dd,J=11.8,2.1Hz,1H),4.32(dd,J=11.8,5.2Hz,1H),4.24–4.18(m, 1H),4.13(d,J=9.5Hz,1H),4.03(t,J=2.7Hz,1H),2.77–2.73(m,2H),2.62(t,J=6.4Hz,2H),2.16(s,3H); 13C NMR (100MHz, CDCl3) δ207.33,172.53,165.56,137.70,137.51,133.15,129.80,129.68,128.59,128.42,128.28,128.2 2,127.97,127.73,127.64,92.49,76.24,74.74,74.03,73.43,72.94,69.58,63.47,37.93,29.74,27.93; HRMS(ESI)m / z C 32 H 34 NaO9(M+Na) + Theoretical value is 585.2095, measured value is 585.2098.

[0064]

[0065] (16) Compound 12 (517.6 mg, 0.92 mmol) was dissolved in anhydrous dichloromethane (2 mL), Cs2CO3 (89.9 mg, 0.3 mmol) was added, and redistilled trichloroacetonitrile (0.3 mL, 3.3 mmol) was added dropwise. After reacting at room temperature for 3 h, the reaction was monitored by TLC plate. The mixture was filtered and concentrated in vacuo to obtain monosaccharide module B-1.

[0066]

[0067] (17) Using D-mannose as the raw material, the monosaccharide module C-1 was synthesized according to the method described in the literature “Silvia, V. et al. J. Org. Chem. 2008, 4496–4503.”

[0068]

[0069] Step 2: Synthesis of double-chain fluorinated carrier-protected mannopentaose

[0070] (1) Anhydrous dichloromethane (3 mL) and dichloromethane were added to the monosaccharide module C-1 (480 mg, 0.75 mmol) and the double-chain fluorocarrier D-1 (299 mg, 0.34 mmol, synthesized according to the method in the literature "Feng, YL. Chai, YH. et al. Chin. Chem. Lett. 2017, 1693-1700"). Molecular sieves, and after cooling at -30 ° C for 10 min, a dichloromethane solution of trimethylsilyl trifluoromethanesulfonate (TMSOTf) (0.6 mL, 10 μL trimethylsilyl trifluoromethanesulfonate dissolved in 1 mL dichloromethane) was added dropwise. After reacting for 30 min, the complete reaction was monitored by TLC plate. The reaction solution was quenched with triethylamine (15 μL), filtered, and the filtrate was concentrated. Column chromatography (ethyl acetate / petroleum ether = 1 / 4, v / v) was used to obtain a colorless oily syrup, namely monosaccharide E-1 (395 mg, 0.29 mmol, yield 84%), and the structural characterization data were as follows: 1 H NMR (400MHz, CDCl3) δ7.41–7.19(m,15H),7.15(dd,J=7.1,2.2Hz,2H),6.92(d,J=8.6Hz ,2H),5.40(dd,J=3.2,1.8Hz,1H),4.91(d,J=1.6Hz,1H),4.85(d,J=10.7Hz,1H),4.71( d,J=4.2Hz,1H),4.68(d,J=3.1Hz,1H),4.63(d,J=11.5Hz,1H),4.56–4.40(m,5H),4.01 (dd,J=8.8,3.3Hz,1H),3.93–3.64(m,12H),2.40(tt,J=18.7,6.6Hz,4H),2.14(s,3H); 13 C NMR (100MHz, CDCl3) δ170.45,157.60,138.32,138.23,137.98,129.88,128.37,128.32,128.04,127.93,127.78,127.70,127.66,127 .59,116.11,96.79,78.35,77.20,76.25,75.25,74.38,73.47,71.83,71.58,69.66,68.89,68.79,68.77,63.47(t),31.43(t),21.09.

[0071]

[0072] (2) Monosaccharide E-1 (395 mg, 0.29 mmol) was dissolved in dry tetrahydrofuran (3 mL), and a 0.16 mol / L sodium methoxide methanol solution (0.3 mL) was added to the solution under stirring at room temperature. After reacting for 2 h, the reaction was monitored by TLC plate to complete the reaction. The reaction solution was quenched with an aqueous ammonium chloride solution and extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to obtain a colorless oily viscous syrup, i.e., monosaccharide receptor F-1 (370 mg, 0.28 mmol, yield 98%). The structural characterization data are 1H NMR (400MHz, CDCl3) δ7.41–7.21(m,15H),7.20–7.14(m,2H),6.92(d,J=7.3Hz,2H),4.97(s,1H),4.82(d,J=10.7 Hz,1H),4.72–4.61(m,4H),4.59–4.38(m,4H),4.05(s,1H),3.96–3.64(m,13H),2.49(s,1H),2.48–2.25(m,4H); 13 C NMR (100MHz, CDCl3) δ157.57,138.27,137.94,130.23,129.85,128.51,128.35,128.32,127.96,127.89,127.82,127.78,127.70 ,127.56,116.11,98.10,80.34,77.20,76.30,75.19,74.39,73.48,72.02,71.29,69.68,69.00,68.54,68.45,63.47,31.45(t).

[0073]

[0074] (3) Anhydrous dichloromethane (8.2 mL) and monosaccharide module A-1 (248 mg, 0.42 mmol) and monosaccharide acceptor F-1 (370 mg, 0.28 mmol) were added. Molecular sieves were added, and after cooling at -30°C for 10 min, a dichloromethane solution of trimethylsilyl trifluoromethanesulfonate (0.01 mL, 0.056 mmol) was added dropwise. After reacting for 30 min, the reaction was completed after monitoring by TLC plate. The reaction solution was quenched with triethylamine (20 μL), and the filtrate was concentrated and separated using polytetrafluoroethylene particles to obtain disaccharide 13 (429.3 mg, 0.23 mmol, yield 82%). The structural characterization data are as follows: 1H NMR (400MHz, CDCl3) δ7.94(d,J=7.9Hz,2H),7.30–7.05(m,30H),6.81(d,J=8.4Hz,2H),5.29(d,J=5.7Hz,1H),5.06 (s,1H),4.88(s,1H),4.76(d,J=10.8Hz,1H),4.65(d,J=11.2Hz,1H),4.58(d,J=13.2Hz,2H),4.48(dd,J=19.6,10. 7Hz,5H),4.36(d,J=30.0Hz,5H),4.20(d,J=11.3Hz,1H),4.01(d,J=7.7Hz,2H),3.94(s,1H),3.82(dd,J=21.8,8.0 Hz,2H),3.77–3.74(m,1H),3.73–3.60(m,11H),2.61–2.38(m,4H),2.32(td,J=18.8,16.5,7.1Hz,4H),2.04(s,3H); 13 C NMR (100MHz, CDCl3) δ206.20,171.73,166.26,157.45,138.60,138.32,138.25,138.23,137.8 3,132.90,130.32,130.03,129.71,129.60,128.35,128.24,128.21,128.09,127.84,127.75,1 27.70,127.64,127.48,127.30,116.03,80.04,76.68,76.18,76.01,74.61,73.91,73.37,73.28,72.65,72.60,72.22,70.30,69.57,69.36,68.57,63.70,63.44,37.82,31.41,29.75,28.06.

[0075]

[0076] (4) Disaccharide 13 (429.3 mg, 0.23 mmol) was dissolved in anhydrous dichloromethane (2.9 mL), and then a 1 mol / L methanol solution of hydrazine acetate (CH3COON2H5) (MeOH, 0.35 mL) was added. After reacting at room temperature for 3 h, the reaction was monitored by TLC plate until complete, and the mixture was directly concentrated in vacuo and separated using polytetrafluoroethylene particles to obtain disaccharide receptor 14 (382.3 mg, 0.22 mmol, yield 94%). The structural characterization data are as follows: 1H NMR (400MHz, CDCl3) δ8.04(d,J=7.8Hz,2H),7.36–7.27(m,20H),7.22(q,J=8.2,7.6Hz,10H),6.92(d,J=8.3Hz,2H),5.25(s, 1H),4.97–4.90(m,2H),4.86(d,J=10.7Hz,1H),4.73–4.68(m,2H),4.62(s,2H),4.58(dd,J=7.8,4.1Hz,3H),4.54(d,J=5.3H z,2H),4.49(t,J=4.9Hz,1H),4.41(d,J=11.8Hz,1H),4.29(dd,J=11.5,7.5Hz,2H),4.13(s,2H),4.03(d,J=8.2Hz,1H),3.94 (d,J=2.9Hz,1H),3.90(d,J=9.4Hz,1H),3.84(d,J=3.5Hz,1H),3.81–3.77(m,6H),3.71(q,J=4.8Hz,6H),2.42–2.36(m,4H).

[0077]

[0078] (5) Anhydrous dichloromethane (6.5 mL) and dichloromethane were added to monosaccharide module A-1 (232.7 mg, 0.33 mmol) and disaccharide acceptor 14 (382.3 mg, 0.22 mmol). Molecular sieves were added, and after cooling at -30°C for 10 min, a dichloromethane solution of trimethylsilyl trifluoromethanesulfonate (8 μL, 0.04 mmol) was added dropwise. After reacting for 30 min, the reaction was completed by monitoring the TLC plate. The reaction solution was quenched with triethylamine (20 μL), filtered and the filtrate was concentrated, and then separated using polytetrafluoroethylene particles to obtain trisaccharide 15 (422.3 mg, 0.18 mmol, yield 83%). The structural characterization data are as follows: 1H NMR(400MHz,Chloroform-d)δ8.03(d,J=7.8Hz,2H),7.92(d,J=7.7Hz,2H),7.49(t,J=8.9Hz,2H),7.36–7.26(m,15H),7.20(dd,J=16.2,7.2Hz,26H),6.91(d,J=8.3Hz,2H),5.46(d,J=8.9Hz,1H),5.22(s,2H),4.96(s,1H),4.87(d,J=10.6Hz,2H),4.59(tp,J=31.6,9.6,9.1Hz,14H),4.38(s,2H),4.34(s,1H),4.30(d,J=9.7Hz,2H),4.25(d,J=9.5Hz,2H),4.10(s,2H),4.05–3.99(m,2H),3.94(d,J=13.9Hz,4H),3.82–3.76(m,6H),3.71(s,4H),2.78–2.48(m,4H),2.44–2.33(m,4H),2.13(s,3H); 13 CNMR(100MHz,CDCl3)δ206.22,172.02,166.31,166.09,157.45,138.23,138.11,132.92,132.84,129.69,129.66,128.45,128.38,128.33,128.28,128.23,128.02,127.79,127.70,127.65,127.53,127.41,127.36,127.23,115.99,98.11,79.96,77.00,76.10,75.26,75.07,74.80,74.62,74.29,74.15,73.23,72.72,72.59,72.46,71.93,71.50,70.74,70.44,69.52,69.23,63.42,37.74,31.56,31.34,31.13,29.78,29.68,28.04。

[0079]

[0080] (6) The trisaccharide 15 (422.3 mg, 0.18 mmol) was dissolved in anhydrous dichloromethane (2.3 mL), and then a 1 mol / L methanol solution of hydrazine acetate (0.27 mL) was added. After reacting at room temperature for 3 h, the reaction was monitored by TLC plate until complete, and the mixture was directly concentrated in vacuo and separated using polytetrafluoroethylene particles to obtain the trisaccharide receptor 16 (350.8 mg, 0.16 mmol, yield 88%). The structural characterization data are as follows: 1 H NMR(400MHz,Chloroform-d)δ8.04(d,J=7.3Hz,2H),7.92(d,J=7.5Hz,2H),7.51(q,J=7.3Hz,2H),7.32–7.27(m,15H),7.22(dd,J=1 4.8,8.4Hz,26H),6.91(d,J=8.5Hz,2H),5.29(s,1H),5.23(s,1H),4.98(s,1H),4.88(d,J=4.2Hz,1H),4.85(d,J=4.6Hz,1H),4.80(d ,J=11.5Hz,1H),4.72–4.67(m,2H),4.58(d,J=11.2Hz,5H),4.53(s,1H),4.49–4.43(m,5H),4.38(s,1H),4.36(s,1H),4.31(dd,J=11 .5,4.9Hz,3H),4.10(t,J=18.4Hz,6H),3.92(s,2H),3.83(d,J=5.1Hz,2H),3.79–3.76(m,4H),3.73–3.68(m,8H),2.42–2.34(m,4H).

[0081]

[0082] (7) Anhydrous dichloromethane (4.7 mL) and 1% ethyl acetate were added to the monosaccharide module A-1 (169.2 mg, 0.24 mmol) and the trisaccharide receptor 16 (350.8 mg, 0.16 mmol). Molecular sieves were added, and after cooling at -30°C for 10 min, a dichloromethane solution of trimethylsilyl trifluoromethanesulfonate (6 μL, 0.03 mmol) was added dropwise. After reacting for 30 min, the reaction was completed by monitoring on a TLC plate. The reaction solution was quenched with triethylamine (20 μL), filtered and the filtrate was concentrated, and then separated using polytetrafluoroethylene particles to obtain tetrasaccharide 17 (413.8 mg, 0.15 mmol, yield 93%). The structural characterization data are as follows: 1H NMR(400MHz,Chloroform-d)δ8.03(d,J=7.7Hz,2H),7.95(d,J=8.6Hz,2H),7.52–7.48(m,2H),7.31(q,J=6.6Hz,14H),7.19(dd,J=12.9,6.4Hz,42H),6.90(d,J=8.2Hz,2H),5.44(dd,J=9.6,3.0Hz,1H),5.30(s,1H),5.21(d,J=24.8Hz,2H),4.97(s,1H),4.83(dd,J=20.8,9.3Hz,2H),4.70–4.54(m,10H),4.51–4.35(m,10H),4.32–4.12(m,12H),4.04–3.88(m,8H),3.86–3.66(m,14H),2.70–2.35(m,8H),2.12(s,3H); 13 C NMR(100MHz,CDCl3)δ206.19,172.06,166.27,166.14,166.07,157.44,138.35,138.27,138.21,138.13,137.95,137.81,137.75,132.93,129.69,129.67,128.49,128.46,128.34,128.29,128.20,128.01,127.80,127.69,127.61,127.54,127.48,127.43,127.38,127.25,127.18,127.08,116.00,98.19,80.02,77.98,77.32,76.11,75.29,75.18,74.86,74.53,74.17,73.82,73.23,72.67,72.50,71.92,71.53,70.68,70.63,70.53,69.52,69.25,68.61,63.42,63.14,63.03,37.73,31.35,29.79,29.68,28.03。

[0083]

[0084] (8) Tetrasaccharide 17 (413.8 mg, 0.15 mmol) was dissolved in anhydrous dichloromethane (1.9 mL), and then a 1 mol / L methanol solution of hydrazine acetate (0.23 mL) was added. After reacting at room temperature for 3 h, the reaction was monitored by TLC plate until complete, and the mixture was directly concentrated in vacuo and separated using polytetrafluoroethylene particles to obtain tetrasaccharide receptor G-1 (367.2 mg, 0.14 mmol, yield 92%). The structural characterization data are as follows: 1 H NMR (400MHz, Chloroform-d) δ8.02(d,J=7.4Hz,2H),7.94(t,J=8.4Hz,4H),7.51(dt,J=16.3,7.0Hz,3H),7.36–7.29(m,14H),7.20(dd,J=14. 5,8.6Hz,38H),6.90(d,J=8.4Hz,2H),5.26(s,4H),4.97(s,1H),4.90–4.84(m,2H),4.81(d,J=9.2Hz,1H),4.75(d,J=12.3Hz,2H),4.70(d,J=1 1.3Hz,2H),4.61(d,J=10.3Hz,4H),4.57–4.56(m,2H),4.47(t,J=4.3Hz,4H),4.38(d,J=8.9Hz,4H),4.31–4.24(m,8H),4.16–4.09(m,4H),4. 02(s,2H),3.95(d,J=10.0Hz,2H),3.88(s,2H),3.83(d,J=3.7Hz,2H),3.78(q,J=3.1Hz,4H),3.73–3.68(m,8H),2.37(dd,J=12.4,6.6Hz,4H).

[0085]

[0086] (9) Anhydrous dichloromethane (4.1 mL) was added to the monosaccharide module B-1 (148.1 mg, 0.21 mmol) and the tetrasaccharide acceptor G-1 (367.2 mg, 0.14 mmol). Molecular sieves, and after cooling at -30 ° C for 10 minutes, trifluoromethanesulfonic acid (TfOH, 6 μL, 0.03 mmol) was added dropwise. After reacting for 30 minutes, the complete reaction was monitored by TLC plate. The reaction solution was quenched with triethylamine (20 μL), filtered and concentrated, and then separated using polytetrafluoroethylene particles to obtain double-chain fluorine carrier protected mannopentasaccharide H-1 (376.9 mg, 0.12 mmol, yield 84%). The structural characterization data are as follows: 1H NMR(400MHz,Chloroform-d)δ8.03(d,J=7.8Hz,4H),7.95(d,J=7.8Hz,4H),7.59(t,J=7.5Hz,2H),7.54–7.47(m,2H),7.43(d,J=7.7Hz,2H),7.31(dd,J=12.2,4.5Hz,14H),7.20(dt,J=12.4,6.3Hz,39H),7.16–7.10(m,6H),7.07(s,4H),6.87(dd,J=22.4,8.6Hz,2H),5.60(dd,J=9.6,2.9Hz,1H),5.31(s,2H),5.22(d,J=31.2Hz,2H),4.97(s,1H),4.88(d,J=10.8Hz,1H),4.83(d,J=11.3Hz,2H),4.81–4.73(m,1H),4.71(s,1H),4.68(s,1H),4.65–4.51(m,10H),4.53–4.46(m,2H),4.44–4.34(m,6H),4.31(d,J=11.9Hz,4H),4.23(dd,J=12.8,5.7Hz,10H),4.17–4.10(m,3H),3.99–3.88(m,10H),3.84(d,J=5.8Hz,2H),3.82–3.73(m,4H),3.71(dd,J=10.5,5.5Hz,4H),2.53(d,J=5.6Hz,2H),2.51–2.46(m,2H),2.39(ddt,J=18.5,12.5,6.7Hz,4H),2.03(s,3H); 13C NMR (100MHz, CDCl3) δ206.37,172.20,166.24,166.07,165.52,157.39,138. 28,138.17,138.11,138.06,137.90,137.78,137.73,137.63,133.15,132.9 2,132.87,129.66,128.50,128.43,128.36,128.27,128.20,128.16,128.09,128.00,127.80,127.67,127.56,127.52,127.48,127.44,127.37,127.29,1 27.20,127.09,126.99,115.93,100.09,99.07,98.11,80.44,79.99,79.85,78.08,77.86,76.25,75.98,75.29,75.11,74.67,74.28,74.22,73.95,73.6 8,73.18,72.63,72.55,71.82,71.43,70.73,70.44,70.24,69.42,69.15,68 .56,63.37,62.98,37.63,31.46,31.25,31.03,29.67,27.73; HRMS(ESI)m / zC 166 H 158 F 26 Na2O 35 2+ (M+2Na) 2+ Theoretical value is 1625.4977, measured value is 1625.5006.

[0087]

[0088] Step 3: Synthesis of sulfonated pentasaccharides and their precursors

[0089] (1) The double-chain fluorinated carrier protected mannopentasaccharide H-1 (241.8 mg, 0.075 mmol) was dissolved in anhydrous dichloromethane (0.95 mL), and then a 1 mol / L methanol solution of hydrazine acetate (0.12 mL) was added. After reacting at room temperature for 3 h, the reaction was monitored by TLC plate. The mixture was directly concentrated in vacuo and separated using polytetrafluoroethylene particles to obtain pentasaccharide 18 (230.0 mg, 0.074 mmol, yield 99%). The structural characterization data are as follows: 1H NMR (400MHz, Chloroform-d) δ8.01(t,J=8.3Hz,4H),7.94(t,J=8.6Hz,4H),7.57(t,J=7.3Hz,1H),7.50(d,J=7.1Hz,2H),7.44(d,J=7.0Hz,2H),7. 35–7.26(m,12H),7.25–7.12(m,44H),7.11–7.02(m,7H),6.89(d,J=8.5H z,2H),5.59(dd,J=9.5,3.1Hz,1H),5.31(d,J=5.8Hz,1H),5.25(d,J=7.4H z,2H),5.17(s,1H),4.96(s,1H),4.87(d,J=10.6Hz,1H),4.81–4.76(m,2 H),4.74–4.65(m,3H),4.64–4.49(m,10H),4.48–4.29(m,10H),4.28–4.16 (m,10H),4.15–4.06(m,3H),4.02–3.87(m,8H),3.84–3.74(m,6H),3.70( dd,J=10.4,5.4Hz,7H),3.52(s,2H),2.38(ddt,J=19.4,13.9,6.9Hz,4H).

[0090]

[0091] (2) Pentasaccharide 18 (45.5 mg, 0.015 mmol) was added The molecular sieves were mixed, dissolved in anhydrous dichloromethane (0.95 mL), and tetrazole (CH2N 4, 5.1 mg, 0.073 mmol), dibenzyl-N,N-diisopropylphosphoramidite ((BnO)2PNPr 2, 23μL), reacted at room temperature for 2h, placed at -20℃, and after TLC plate monitoring complete reaction, m-chloroperbenzoic acid (m-CPBA, 15.1mg, 0.066mmol) was added. After reacting for 1h, TLC plate monitoring complete reaction, the reaction solution was dissolved in dichloromethane, filtered, washed with saturated NaCl aqueous solution, extracted with dichloromethane, and the organic phase was separated using polytetrafluoroethylene particles to obtain compound 19 (53.4mg, 0.015mmol, yield 99%), the structural characterization data are as follows: 1H NMR(400MHz,Chloroform-d)δ8.01(dd,J=11.8,7.6Hz,4H),7.94(d,J=7.7Hz,4H),7.58(t,J=7.4Hz,1H),7.54–7.47(m,2H),7.42(d,J=7 .7Hz,2H),7.31(dd,J=7.8,4.0Hz,10H),7.24–7.12(m,48H),7.12–7.04(m,9H),6.97(d,J=6.4Hz,5H),6.89(d,J=8.4Hz,2H),5.60(dd,J =9.9,3.1Hz,1H),5.26(d,J=7.7Hz,2H),5.12(s,1H),4.97(s,1H),4.95–4.85(m,5H),4.82(d,J=10.3Hz,2H),4.72(s,2H),4.63–4.50(m ,11H),4.49–4.28(m,10H),4.27–4.14(m,12H),4.13–3.98(m,6H),3.97–3.81(m,11H),3.8–3.63(m,11H),2.38(dd,J=16.3,9.5Hz,4H).

[0092]

[0093] (3) Compound 19 (49.4 mg, 0.015 mmol) was dissolved in dry methanol (0.15 mL), and freshly prepared sodium methoxide solution (prepared by dissolving 0.34 mg Na in 0.1 mL methanol) was added. After reacting at room temperature for 3 h, the reaction was completed after monitoring by TLC plate. The pH was adjusted to 7 using Amberlite (R) IRC 120H resin, and then separated using polytetrafluoroethylene particles to obtain compound 20 (39 mg, 0.013 mmol, yield 87%). The structural characterization data are as follows: 11H NMR (400 MHz, Chloroform-d) δ 8.10 (d, J = 7.7 Hz, 2H), 7.61 (t, J = 7.5 Hz, 1H), 7.47 (t, J = 7.7 Hz, 2H), 7.37 (d, J = 7.5 Hz, 3H), 7.29 (d, J = 4.7 Hz, 8H), 7.24 (d, J = 3.8 Hz, 8H), 7.22 (s, 8H), 7.21 (d, J = 3.4 Hz, 13H), 7.18 (s, 7H), 7.16 (s, 5H), 7.15 (s, 3H), 7.13–7.06 (m, 7H), 6.91 (d, J = 8.3 Hz, 2H), 5.30 (s, 1H), 5.19 (s, 3H), 4.96–4.88 (m, 5H), 4.80 (dd, J = 10.9, 6.2 Hz, 3H), 4.69 (dd, J = 8.5, 4.9 Hz, 4H), 4.64 (s, 2H), 4.61 (s, 2H), 4.58 (d, J = 12.1 Hz, 2H), 4.52 (d, J = 11.0 Hz, 2H), 4.47 (d, J = 10.2 Hz, 3H), 4.40 (d, J = 11.6 Hz, 2H), 4.36 (s, 1H), 4.28–4.20 (m, 4H), 4.15–4.09 (m, 4H), 4.05 (s, 2H), 4.01 (d, J = 5.2 Hz, 1H), 3.95 (d, J = 6.6 Hz, 2H), 3.91 (d, J = 2.5 Hz, 1H), 3.89 (s, 2H), 3.85 (d, J = 9.9 Hz, 2H), 3.77 (dq, J = 6.8, 3.6 Hz, 8H), 3.70 (d, J = 5.2 Hz, 5H), 3.68–3.64 (m, 7H), 3.61 (d, J = 9.9 Hz, 5H), 3.51 (dd, J = 10.5, 4.5 Hz, 2H), 2.39 (ddd, J = 18.7, 11.8, 6.5 Hz, 4H); HRMS (ESI) m / z C 147 H 149 F 26 Na2O 32 P 2+ (M + 2Na) 2+ Calculated 1498.4570, Found 1498.4569.

[0094]

[0095] (4) Compound 20 (55.8 mg, 0.019 mmol) was dissolved in a mixture of methanol, water, and ethyl acetate in a volume ratio of 4:1:3 (4.4 mL:1.1 mL:3.3 mL), and Pd(OH)2 / C (46.5 mg) was added under ice bath. The mixture was reacted at room temperature for 72 h under H2 atmosphere. After the reaction was completed as monitored by TLC plate, the mixture was extracted with water to obtain phosphomannopentose P-1 (15.2 mg, 0.017 mmol, yield 89%). The structural characterization data are as follows: 1 H NMR(400MHz,D2O)δ5.37(s,1H),5.11(s,2H),5.08(s,1H),5.02(s,1H),4.32(s,1H),4.21(s,3H),4.06(s,2H ), 3.95 (d, J = 8.7Hz, 6H), 3.86 (d, J = 12.2Hz, 6H), 3.78 (d, J = 6.8Hz, 6H), 3.72 (d, J = 9.9Hz, 6H); HRMS (ESI) m / z C 30 H 52 O 29 P - (MH) - Theoretical value is 907.2337, measured value is 907.2334.

[0096]

[0097] (5) Phosphomannopentose P-1 (14.9 mg, 0.005 mmol) and sulfur trioxide pyridinium salt complex (SO3·Et3N, 79.6 mg, 0.5 mmol) were dissolved in anhydrous triethylamine (0.05 mL) and pyridine (0.5 mL), and microwaved at 120°C for 40 min. Methanol (0.3 mL) was added and stirred for 5 min. Amberlite IR 120 (Na) was added and separated and purified by Sephadex G-25 gel exclusion chromatography to obtain sulfonated mannopentose S-1 (8.6 mg, 79%). The structural characterization data are as follows: 1 H NMR(400MHz,D2O)δ5.81(s,1H),5.55(s,1H),5.48(s,1H),5.46–5.38(m,2H),5.23(s,1H),4.99(S,1H ),4.88–4.80(m,2H),4.77–4.73(m,2H),4.70–4.46(m,8H),4.44–4.17(m,14H),4.11(d,J=9.8Hz,1H); 13C NMR (100MHz, CDCl3) δ101.34,99.84,99.80,98.24,97.11,76.99,76.53,76.38,76.10,75.98,75.46,75.00,74.92,74.51,74 .04,72.66,72.38,72.22,71.71,71.53,71.30,70.18,69.59,69.25,69.15,66.86,66.45,66.01,65.87,62.88; HRMS(ESI)m / z C 30 H 36 O 77 Na 11 PS 16 [M+H+11Na] 6- Theoretical value: 403.8841, measured value: 403.8022.

[0098]

[0099] Example 2

[0100] Step 1: Synthesis of monosaccharide modules

[0101] This step is the same as step 1 in Example 1.

[0102] Step 2: Synthesis of double-chain fluorinated carrier-protected mannotetraose

[0103] The trisaccharide receptor 16 was synthesized according to the steps (1) to (6) in step 2 of Example 1.

[0104] (7) Anhydrous dichloromethane (1.1 mL) was added to the monosaccharide module B-1 (37.7 mg, 0.053 mmol) and the trisaccharide acceptor 16 (80.8 mg, 0.036 mmol). Molecular sieves were added, and after cooling at -30°C for 10 min, trifluoromethanesulfonic acid (0.4 μL, 0.005 mmol) was added dropwise. After reacting for 30 min, the reaction was completed after monitoring by TLC plate. The reaction solution was quenched with triethylamine (5 μL), filtered and the filtrate was concentrated, and then separated using polytetrafluoroethylene particles to obtain double-chain fluorine carrier-protected mannotetraose H-2 (132.4 mg, 0.048 mmol, yield 91%). The structural characterization data are as follows: 1H NMR(400MHz,Chloroform-d)δ7.96(d,J=7.8Hz,4H),7.85(d,J=7.7Hz,2H),7.50(t,J=7.5Hz,1H),7.42(dt,J=14.2,7.5Hz,2H),7.36(t,J=7.7Hz,2H),7.23(d,J=8.7Hz,5H),7.12(ddt,J=31.1,14.1,6.7Hz,41H),6.99(d,J=4.4Hz,4H),6.82(d,J=8.1Hz,2H),5.54(d,J=9.5Hz,1H),5.26(s,1H),5.16(d,J=16.8Hz,2H),4.90(s,1H),4.79(dd,J=16.9,11.1Hz,2H),4.72(d,J=10.6Hz,1H),4.62(d,J=10.8Hz,2H),4.53(t,J=11.5Hz,5H),4.48–4.43(m,3H),4.42–4.36(m,4H),4.34–4.28(m,3H),4.27–4.13(m,9H),4.09(d,J=11.2Hz,1H),4.06–4.02(m,2H),3.94(d,J=10.0Hz,1H),3.91(s,2H),3.89–3.82(m,5H),3.75(d,J=6.6Hz,2H),3.70(q,J=6.8Hz,4H),3.62(ddd,J=19.7,10.3,4.3Hz,6H),2.48(t,J=6.6Hz,2H),2.42(t,J=6.6Hz,2H),2.32(tt,J=18.8,6.7Hz,4H),1.98(s,3H); 13C NMR (100MHz, CDCl3) δ206.39,172.27,166.25,166.11,165.57,157.40,138.2 9,138.21,138.15,138.11,137.80,137.70,133.15,132.92,132.85,130.19, 129.94,129.85,129.74,129.67,129.64,128.49,128.44,128.37,128.32,128.27,128.24,128.19,128.10,128.01,127.83,127.79,127.73,127.69,127.5 9,127.56,127.47,127.38,127.29,127.16,115.96,100.03,98.18,80.00,77.85,77.65,76.39,76.04,75.30,75.15,74.77,74.67,74.29,73.99,73.21,7 2.84,72.62,72.57,71.85,71.63,70.60,70.28,69.47,69.44,69.15,68.59, 63.39,63.04,63.01,37.67,31.44,31.30,31.15,29.69,27.78; HRMS(ESI)m / z C 139 H 132 F 26 Na2O 29 P 2+ (M+Na) 2+ Theoretical value: 1402.4112; measured value: 1402.4127.

[0105]

[0106] Step 3: Synthesis of sulfonated tetrasaccharide precursor

[0107] The double-chain fluorinated carrier protected mannopentasaccharide H-2 (169.1 mg, 0.061 mmol) was dissolved in anhydrous dichloromethane (0.8 mL), and then a 1 mol / L methanol solution of hydrazine acetate (0.09 mL) was added. After reacting at room temperature for 3 h, the reaction was monitored by TLC plate, and then directly concentrated in vacuo, and then separated using polytetrafluoroethylene particles to obtain compound 21 (159.6 mg, 0.06 mmol, yield 99%). The structural characterization data are as follows: 1H NMR(400MHz,Chloroform-d)δ8.02(dd,J=7.8,4.5Hz,4H),7.92(d,J=7.7Hz,2H),7.58(t,J =7.4Hz,1H),7.50(d,J=7.3Hz,2H),7.43(t,J=7.7Hz,3H),7.31(t,J=6.9Hz,10H),7.22(q, J=7.4,5.9Hz,23H),7.17(t,J=7.2Hz,12H),7.10–7.04(m,5H),6.87(dd,J=22.1,8.4Hz,2H ),5.61(dd,J=9.6,3.1Hz,1H),5.30(d,J=4.1Hz,1H),5.25(s,1H),5.20(s,1H),4.96(s,1H) ,4.87(d,J=10.8Hz,1H),4.84–4.66(m,5H),4.61–4.56(m,6H),4.53(d,J=6.6Hz,2H),4.46 (p,J=6.7,5.9Hz,5H),4.38(d,J=5.0Hz,3H),4.31(s,1H),4.26(q,J=6.6,5.1Hz,6H),4.12( d,J=9.1Hz,2H),4.03–3.93(m,5H),3.91(d,J=14.5Hz,2H),3.83(d,J=6.9Hz,2H),3.78(tt ,J=6.7,3.4Hz,5H),3.69(td,J=9.9,4.4Hz,6H),3.54(s,2H),2.39(tt,J=19.1,6.8Hz,4H).

[0108]

[0109] (3) Tetrasaccharide 21 (77.2 mg, 0.029 mmol) was added The mixture was mixed with 2-nitrogen-1-yl molecular sieves, dissolved in anhydrous dichloromethane (2.9 mL), tetrazole (10.2 mg, 0.073 mmol) and dibenzyl-N,N-diisopropylphosphoramidite (39 μL) were added, and the mixture was reacted at room temperature for 2 h, and then placed at -20 ° C. After the TLC plate monitored the complete reaction, m-chloroperbenzoic acid (26.7 mg, 0.12 mmol) was added and reacted for 1 h. After the TLC plate monitored the complete reaction, the reaction solution was dissolved in dichloromethane, filtered, washed with saturated NaCl aqueous solution, extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate and filtered. After vacuum concentration, it was separated using polytetrafluoroethylene particles to obtain compound 22 (81.78 mg, 0.028 mmol, yield 99%), and the structural characterization data were as follows: 1H NMR(400MHz,Chloroform-d)δ8.02(dd,J=11.5,7.7Hz,4H),7.91(d,J=7.8Hz,2H),7.58(t,J=7.5 Hz,1H),7.48(t,J=4.1Hz,2H),7.44(t,J=7.8Hz,3H),7.35–7.27(m,10H),7.25–7.21(m,20H),7. 20–7.17(m,15H),7.13(d,J=6.8Hz,5H),7.10–7.03(m,5H),6.98(d,J=4.3Hz,5H),6.86(dd,J=22 .6,8.5Hz,2H),5.62(dd,J=9.9,3.0Hz,1H),5.30(s,1H),5.26(s,1H),5.17(s,1H),4.98–4.91(m, 5H),4.89(d,J=4.2Hz,1H),4.84(d,J=11.7Hz,2H),4.74(d,J=11.3Hz,1H),4.69(d,J=11.1Hz,1H ),4.63(d,J=6.5Hz,1H),4.59(dd,J=7.1,4.4Hz,3H),4.57–4.52(m,5H),4.46(dd,J=9.3,3.8Hz,3 H),4.38(t,J=9.9Hz,4H),4.27(dd,J=10.7,7.7Hz,5H),4.19(d,J=9.6Hz,3H),4.12(s,1H),4.03 (s,2H),3.98–3.91(m,5H),3.88(s,2H),3.84–3.66(m,13H),2.39(ddt,J=18.9,13.7,6.8Hz,4H).

[0110]

[0111] (4) Compound 22 (95.9 mg, 0.033 mmol) was dissolved in dry methanol (0.3 mL), and freshly prepared sodium methoxide (prepared by dissolving 0.8 mg Na in 0.1 mL MeOH) was added. After reacting at room temperature for 3 h, the reaction was completed after monitoring by TLC plate. The pH was adjusted to 7 using Amberlite (R) IRC 120H resin, and the mixture was filtered and concentrated in vacuo. Polytetrafluoroethylene particles were used for separation to obtain compound 23 (60.3 mg, 0.023 mmol, yield 70%). The structural characterization data are as follows: 1H NMR(400MHz,D2O)δ7.37(d,J=7.4Hz,4H),7.28(d,J=10.3Hz,13H),7.25–7.08(m,40H),6.91(d,J=8.0Hz,2H),5.22(s,2H), 5.19(s,1H),4.96(d,J=5.5Hz,3H),4.92(d,J=10.1Hz,2H),4.81(d,J=10.9Hz,2H),4.74–4.59(m,7H),4.59(s,2H),4.59–4. 49(m,4H),4.47(s,2H),4.43(d,J=3.5Hz,2H),4.30–4.19(m,4H),4.09(d,J=15.4Hz,5H),3.99(s,2H),3.96–3.84(m,4H),3. 82–3.74(m,7H),3.74(s,3H),3.73–3.60(m,11H),3.50(dd,J=11.1,4.2Hz,1H),2.39(td,J=18.6,9.2Hz,4H); HRMS(ESI)m / z C 127 H 127 F 26 NaO 27 P + (M+Na) + Theoretical value is 2631.7779, measured value is 2631.7773.

[0112]

[0113] (5) Compound 23 (60.3 mg, 0.023 mmol) was dissolved in a mixture of methanol, water, and ethyl acetate in a volume ratio of 4:1:3 (4.4 mL:1.1 mL:3.3 mL), and 20% Pd(OH)2 / C (46.5 mg) was added under ice bath. After reacting at room temperature for 72 h under H2 atmosphere, the reaction was completed after monitoring by TLC plate, and then extracted with water to obtain phosphomannotetraose P-2 (14.1 mg, 0.019 mmol, yield 83%). The structural characterization data are as follows: 1 H NMR(400MHz,D2O)δ5.36(s,1H),5.10(s,1H),5.06(s,1H),5.00(s,1H),4.30(s,1H),4.1 6(d,J=18.1Hz,2H),4.05(s,1H),3.94–3.81(m,10H),3.78–3.64(m,10H); HRMS(ESI)m / z C 24 H 42 O 24 P - (MH)- Theoretical value: 745.1809, measured value: 745.1814.

[0114]

Claims

1. A method for synthesizing an anti-tumor and anti-angiogenic drug PI-88 and its precursor, characterized in that The steps include: Step 1: Synthesis of monosaccharide modules Using D-mannose as a raw material, compound I is obtained by protecting its hydroxyl position, and after removing the R1 protecting group at position 1 of compound I, it is reacted with trichloroacetonitrile to obtain a monosaccharide module A, as shown in route (1): Using D-mannose as a raw material, compound II is obtained by protecting its hydroxyl position, and after removing the R1 protecting group at position 1 of compound II, it is reacted with trichloroacetonitrile to obtain a monosaccharide module B, as shown in route (2): Using D-mannose as a raw material, compound II is obtained by protecting its hydroxyl position, and after removing the R1 protecting group at position 1 of compound II, it is reacted with trichloroacetonitrile to obtain monosaccharide module C, as shown in route (3): wherein the R1 protecting group is selected from any one of acetyl, ethylthio, p-methylphenylthio, o-iodobenzylthio, and benzylthio; the R2, R3, R4, and R5 protecting groups are each independently selected from any one of acetyl, benzoyl, benzyl, p-methoxybenzyl, tert-butyldiphenylsilyl, tert-butyldimethylsilyl, and trityl; the R6 protecting group is selected from any one of acetyl and acetyl propionyl, and the R6 protecting group is different from the R1, R3, R4, and R5 protecting groups; Step 2: Synthesis of double-stranded fluorophore-protected mannopolysaccharide The double-chain fluorine carrier D and the monosaccharide module C are glycosylated to obtain monosaccharide E, and then the 2-position R6 protecting group is removed to obtain monosaccharide receptor F; the monosaccharide receptor F is repeatedly glycosylated with the monosaccharide module A for n times and then the 3-position Lev protecting group is removed to obtain compound G; compound G is then glycosylated with the monosaccharide module B to obtain double-chain fluorine carrier protected mannopolysaccharide H, as shown in route (4): Wherein, n is an integer from 1 to 3, a is an integer from 0 to 2, and b is an integer from 4 to 7; Step 3: Synthesis of PI-88 and its precursors The Lev protecting group at the 6th position of the terminal sugar unit in the double-stranded fluorine carrier protected mannopolysaccharide H was removed and phosphorylated, and the remaining protecting groups were selectively removed to obtain PI-88 precursors with different numbers of exposed hydroxyl groups; The exposed hydroxyl groups in the PI-88 precursor are sulfonated, and then all other protecting groups are removed to obtain PI-88.

2. The method for synthesizing the anti-tumor and anti-angiogenesis drug PI-88 and its precursor according to claim 1, characterized in that: In step 1, the R1 protecting group is p-methylphenylthio, and the R2, R3, R4, and R5 protecting groups are each independently selected from any one of benzoyl, benzyl, p-methoxybenzyl, tert-butyldiphenylsilyl, tert-butyldimethylsilyl, and trityl; and the R6 protecting group is acetyl.

3. The method for synthesizing the anti-tumor and anti-angiogenesis drug PI-88 and its precursor according to claim 1, characterized in that: In step 2, the method for glycosylation coupling of the double-chain fluoro carrier D and the monosaccharide module C is: the double-chain fluoro carrier D and the monosaccharide module C are mixed and dissolved in a solvent, an acid catalyst is added, and the reaction is carried out at a temperature of -20 to -40°C for 0.5 to 2h; the solvent is selected from any one of dichloromethane, tetrahydrofuran, toluene, and acetonitrile, or a mixture of two of them; the acid catalyst is trimethylsilyl trifluoromethanesulfonate or trifluoromethanesulfonic acid.

4. The method for synthesizing the anti-tumor and anti-angiogenesis drug PI-88 and its precursor according to claim 1, characterized in that: In step 2, the method for glycosylation coupling of the monosaccharide receptor F and the monosaccharide module A is: the monosaccharide receptor F and the monosaccharide module A are mixed and dissolved in a solvent, an acid catalyst is added, and the reaction is carried out at a temperature of -20 to -40°C for 0.5 to 2h; the solvent is selected from any one of dichloromethane, tetrahydrofuran, toluene, and acetonitrile or a mixture of two thereof; the acid catalyst is trimethylsilyl trifluoromethanesulfonate or trifluoromethanesulfonic acid.

5. The method for synthesizing the anti-tumor and anti-angiogenesis drug PI-88 and its precursor according to claim 1, characterized in that: In step 2, the method for removing the 3-position Lev protecting group is: using dichloromethane or a pyridine / acetic acid mixed solution as a solvent, adding a methanol solution of hydrazine acetate, and reacting at room temperature for 2 to 6 hours.

6. The method for synthesizing the anti-tumor and anti-angiogenesis drug PI-88 and its precursor according to claim 1, characterized in that: In step 2, the method for glycosylation reaction of compound G and monosaccharide module B is: compound G and monosaccharide module B are mixed and dissolved in a solvent, an acid catalyst is added, and the reaction is carried out at a temperature of -20 to -40°C for 0.5 to 2h; the solvent is selected from any one of dichloromethane, tetrahydrofuran, toluene, acetonitrile or a mixture of two thereof; the acid catalyst is trimethylsilyl trifluoromethanesulfonate or trifluoromethanesulfonic acid.

7. The method for synthesizing the anti-tumor and anti-angiogenesis drug PI-88 and its precursor according to claim 1, characterized in that: In step 3, the method for removing the Lev protecting group at the 6-position of the terminal sugar unit in the double-stranded fluorine carrier protected mannopolysaccharide H is as follows: dissolving mannopolysaccharide H in dichloromethane or a pyridine / acetic acid mixed solution, then adding a methanol solution of hydrazine acetate, and reacting at room temperature for 2 to 6 hours.

8. The method for synthesizing the anti-tumor and anti-angiogenesis drug PI-88 and its precursor according to claim 1, characterized in that: In step 3, the phosphorylation method is: using dichloromethane as a solvent, adding tetrazole and dibenzyl-N,N-diisopropylphosphoramidite, reacting at room temperature for 0.5 to 2 hours, cooling to -20 to -40°C, adding m-chloroperbenzoic acid and reacting for another 0.5 to 2 hours.

9. The method for synthesizing the anti-tumor and anti-angiogenesis drug PI-88 and its precursor according to claim 1, characterized in that: In step 3, the sulfonation method is: mixing the PI-88 precursor and the sulfur trioxide salt complex, dissolving the mixture in a solvent, and subjecting the mixture to a microwave reaction at 100 to 140° C. for 0.3 to 1 h; or mixing the PI-88 precursor and the sulfur trioxide salt complex, dissolving the mixture in a solvent, and subjecting the mixture to a reaction at 50 to 80° C. for 1 to 3 d; the solvent is selected from any one or two of triethylamine, tetrahydrofuran, pyridine, and N,N-dimethylformamide; and the sulfur trioxide salt complex is selected from any one of sulfur trioxide trimethylamine salt complex, sulfur trioxide triethylamine salt complex, and sulfur trioxide pyridine salt complex.

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