Fucosan sulfate compound as well as preparation method and application thereof

Abstract

The invention provides a fucosan sulfate compound as well as a preparation method and application thereof, and belongs to the technical field of organic chemistry and medicinal chemistry. The anticoagulant activity of the fucosan sulfate compound is high, the anticoagulant activity of the compound 29, the anticoagulant activity of the compound 30, the anticoagulant activity of the compound 37 and the anticoagulant activity of the compound 58 are high, EC2.0 * multiplied by APTT reaches 13.6 mu g / mL, 14.5 mu g / mL, 16.4 mu g / mL and 12.9 mu g / mL respectively, and the activity of the EC2.0 * multiplied by APTT is close to that of an anticoagulant drug enoxaparin commonly used in clinic. However, different from enoxaparin acting on a common blood coagulation pathway, safety problems such as bleeding, allergy and gastrointestinal tract side reactions exist, and fucoidan only acts on an endogenous blood coagulation pathway, so that the risk of bleeding is greatly reduced from the mechanism. In addition, the fucoidan is derived from invertebrates and cannot be degraded by enzymes in mammals, so that oral administration is expected to be realized, and the compliance of patients is greatly improved.

Description

Technical Field

[0001] The present invention relates to the technical fields of organic chemistry and medicinal chemistry, and particularly relates to a class of fucoidan sulfate compounds, a preparation method thereof, and an application thereof. Background Art

[0002] At present, the incidence of cardiovascular and cerebrovascular diseases induced by thrombosis is increasing worldwide, seriously threatening human health. Research shows that the three main factors for the formation of thrombus are slow blood flow, hypercoagulable state of blood, and vascular injury. According to the mechanism of thrombus formation, antithrombotic drugs are divided into three categories, namely anticoagulants, platelet inhibitors, and fibrinolytic agents, with anticoagulants being the main treatment means. Representative drugs of anticoagulants include: 1) low molecular weight heparin, such as enoxaparin sodium; 2) non-peptide thrombin inhibitors, such as dabigatran; 3) oral small molecule coagulation factor inhibitors, such as rivaroxaban, apixaban, etc. Although these drugs have excellent performance in delaying blood coagulation time, since they all act on the common coagulation pathway, they can induce thrombocytopenia, inhibit coagulation factors, etc., so they are prone to bleeding and it is not easy to stop bleeding after bleeding. In addition, long-term use of anticoagulant drugs by patients can damage the gastrointestinal mucosa, leading to gastrointestinal mucosal erosion, ulcers, and accompanied by gastrointestinal reactions such as nausea, vomiting, abdominal pain, and diarrhea. In summary, the bleeding, allergy, and gastrointestinal side effects brought by traditional anticoagulant drugs need to be solved urgently.

[0003] Related technologies have proved that naturally extracted fucoidan sulfate has good anticoagulant activity, and different from traditional anticoagulant drugs acting on the common coagulation pathway, it mainly exerts its anticoagulant effect by inhibiting the intrinsic coagulation pathway. Therefore, fucoidan sulfate can avoid the bleeding problem of traditional anticoagulant drugs and is an important research direction for the development of new anticoagulant drugs. However, naturally extracted fucoidan sulfate has the problem of low anticoagulant activity. Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a class of fucoidan sulfate compounds, a preparation method thereof, and an application thereof. The fucoidan sulfate compounds of the present invention have high anticoagulant activity.

[0005] In order to achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0006] The present invention provides a class of fucoidan sulfate compounds having the structures shown in Formula I or II:

[0007]

[0008] The fucoidan sulfate compounds having the structure shown in Formula I include the structures shown in any one of Formulas 1 to 30, E, F, or G:

[0009]

[0010] The fucoidan sulfate compounds with the structure shown in Formula II include the structures shown in any one of Formulas 31 to 58:

[0011]

[0012] The present invention also provides a preparation method of the fucoidan sulfate compounds described in the above technical solution, including the following steps:

[0013] Using fucose as a raw material to obtain monosaccharide building blocks 1-4;

[0014] Replacing the protecting group at the 3-position of the monosaccharide building blocks 1-4 with an electron-donating group TBS to obtain monosaccharide building blocks 1-6;

[0015] Performing a deprotection reaction on the monosaccharide building blocks 1-6 to obtain monosaccharide building blocks 1-8;

[0016]

[0017] Using the monosaccharide building blocks 1-4, 1-6 and 1-8, based on the one-pot synthesis reaction with pre-activated glycosyl donors, to obtain Type I fucosylated oligosaccharide fragments;

[0018] Performing polymerization on the Type I fucosylated oligosaccharide fragments to obtain Type I fucoidan fragments, and removing the temporary protecting group TBS on the sugar chain of the Type I fucoidan fragments to obtain Type I fucoidan;

[0019] Performing selective deprotection on the Type I fucoidan to obtain a Type I deprotected product;

[0020] Performing sulfation modification on the 2-position of the sugar ring of the Type I deprotected product to obtain Compounds 1 to 11; performing sulfation modification on the 4-position of the sugar ring of the Type I deprotected product to obtain Compounds 12 to 22; performing sulfation modification on both the 2-position and 4-position of the sugar ring of the Type I deprotected product to obtain Compounds 23 to 30 and Compounds E, F and G;

[0021] Forming PMB groups on the monosaccharide building blocks 1-4 and 1-8 respectively to obtain monosaccharide building blocks 1-10 and 1-12;

[0022]

[0023] Using the monosaccharide building blocks 1-6, 1-4, 1-10 and 1-12, based on the one-pot synthesis reaction with pre-activated glycosyl donors, to obtain Type II fucosylated oligosaccharide fragments;

[0024] Polymerize the Type II fucosooligosaccharide fragment to obtain a Type II fucoidan fragment, and remove the TBS temporary protecting group on the sugar chain of the Type II fucoidan fragment to obtain Type II fucoidan;

[0025] Selectively remove the protecting group from the Type II fucoidan to obtain a Type II deprotected product;

[0026] Perform sulfation modification on the 2- and 3-positions of the sugar ring of the Type II deprotected product simultaneously to obtain Compounds 31-41; perform sulfation modification on the 3- and 4-positions of the sugar ring of the Type II deprotected product simultaneously to obtain Compounds 42-52; perform sulfation modification on the 2-, 3-, and 4-positions of the sugar ring of the Type II deprotected product simultaneously to obtain Compounds 53-58.

[0027] Preferably, the temperature for sulfation modification at the 2-position, sulfation modification at the 4-position, simultaneous sulfation modification at the 2- and 4-positions, simultaneous sulfation modification at the 2- and 3-positions, simultaneous sulfation modification at the 3- and 4-positions, and simultaneous sulfation modification at the 2-, 3-, and 4-positions is independently room temperature, and the time is independently 7-14 h.

[0028] Preferably, the sulfating reagents for sulfation modification at the 2-position, sulfation modification at the 4-position, simultaneous sulfation modification at the 2- and 4-positions, simultaneous sulfation modification at the 2- and 3-positions, simultaneous sulfation modification at the 3- and 4-positions, and simultaneous sulfation modification at the 2-, 3-, and 4-positions include sulfur trioxide pyridine (SO3·Pyr) and / or sulfur trioxide trimethylamine (SO3·NMe3).

[0029] Preferably, when performing sulfation modification at the 2-position, sulfation modification at the 4-position, simultaneous sulfation modification at the 2- and 4-positions, simultaneous sulfation modification at the 2- and 3-positions, simultaneous sulfation modification at the 3- and 4-positions, and simultaneous sulfation modification at the 2-, 3-, and 4-positions, the equivalent ratio of sulfur trioxide pyridine to each naked hydroxyl group on the sugar ring in the raw material is 10:1, and the raw material is the Type I deprotected product or the Type II deprotected product.

[0030] Preferably, after the sulfation modification at the 2-position, the sulfation modification at the 4-position, the simultaneous sulfation modification at the 2- and 4-positions, the simultaneous sulfation modification at the 2- and 3-positions, the simultaneous sulfation modification at the 3- and 4-positions, and the simultaneous sulfation modification at the 2-, 3- and 4-positions are completed, the obtained product is further neutralized, and the basic substance used for neutralization includes sodium hydroxide.

[0031] The present invention also provides the application of the fucan sulfate compounds described in the above technical solution in the preparation of anti-coagulant active drugs.

[0032] Preferably, the dosage form of the anti-coagulant active drug includes pharmaceutically acceptable dosage forms such as tablets, injections, capsules, granules, pills, powders, oral liquids, sustained-release preparations, controlled-release preparations or nano-preparations.

[0033] Preferably, the anti-coagulant active drug contains an effective dose of the fucan sulfate compounds, pharmaceutically acceptable salts and pharmaceutically acceptable carriers, excipients, excipients and diluents.

[0034] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0035] The fucan sulfate compounds of the present invention have high anti-coagulant activity, and the compounds A - D have relatively strong anti-coagulant activity. The EC 2.0× for doubling APTT reaches 13.6 μg / mL, 14.5 μg / mL, 16.4 μg / mL and 12.9 μg / mL respectively, which is similar to the activity of the clinically commonly used anticoagulant drug enoxaparin. However, different from enoxaparin that acts on the common coagulation pathway and has safety problems such as bleeding, allergy, and gastrointestinal side reactions, fucan sulfate only acts on the intrinsic coagulation pathway, significantly reducing the risk of bleeding from the mechanism. In addition, since fucan sulfate is derived from invertebrates and will not be degraded by the enzymes in mammalian bodies, it is expected to achieve oral administration, greatly improving the compliance of patients.

[0036] The present invention also provides a preparation method of the fucan sulfate compounds described in the above technical solution. By applying the pre-activated one-pot synthesis strategy to the synthesis of fucan, the synthesis efficiency is higher, the cumbersome protection / deprotection steps are reduced, and the synthesis route is shorter. Description of the Drawings

[0037] Figure 1 It is a preparation process diagram of monosaccharide building blocks 1-4 and 1-5;

[0038] Figure 2 It is a preparation process diagram of monosaccharide building block 1-6;

[0039] Figure 3Process diagram for the preparation of monosaccharide building blocks 1-8;

[0040] Figure 4 Process diagram for the one-pot synthesis of a tetrasaccharide donor for fucoidan;

[0041] Figure 5 Process diagram for the one-pot synthesis of a fucoidan tetrasaccharide fragment 2-3 with a blocked end;

[0042] Figure 6 Process diagram for the preparation of disaccharide receptors 1-13, 1-14, 2-1, 3-1 and tetrasaccharide receptor 1-16;

[0043] Figure 7 Process diagram for the preparation of octasaccharide 2-7;

[0044] Figure 8 Process diagram for the one-pot synthesis of a dodecaccharide fragment;

[0045] Figure 9 Process diagram for the one-pot synthesis of target trisaccharides 2-2, 3-2;

[0046] Figure 10 Process diagram for the one-pot synthesis of target tetrasaccharides 2-3, 3-3;

[0047] Figure 11 Schematic diagram for obtaining fucoidan sulfate compounds 1-11;

[0048] Figure 12 Schematic diagram for obtaining fucoidan sulfate compounds 12-22;

[0049] Figure 13 Schematic diagram for obtaining fucoidan sulfate compounds 23-30 and formulas E, F and G;

[0050] Figure 14 Process diagram for the preparation of monosaccharide building blocks 1-10, 1-12;

[0051] Figure 15 Process diagram for the preparation of fucotetraose 1-19;

[0052] Figure 16 Process diagram for the preparation of fucotetraose 10-3;

[0053] Figure 17 Process diagram for the preparation of glycosyl receptors 1-18, 1-20, 11-1, 11-3;

[0054] Figure 18 Process diagram for the one-pot synthesis of target trisaccharides 10-2, 11-2;

[0055] Figure 19Process diagram for the one-pot synthesis of the target tetrasaccharides 10-3 and 11-3;

[0056] Figure 20 Process diagram for the preparation of compounds 12-1 to 12-11;

[0057] Figure 21 Process diagram for the removal of the PMB protecting group to obtain compounds 13-1 to 13-7;

[0058] Figure 22 Process diagram for the removal of the PMB protecting group to obtain compounds 13-8 to 13-11;

[0059] Figure 23 Process diagram for the removal of the Bz protecting group to obtain compounds 14-1 to 14-11;

[0060] Figure 24 Process diagram for the simultaneous removal of the Bn and Bz protecting groups to obtain compounds 15-1 to 15-6. Detailed implementation mode

[0061] The present invention provides a class of fucoidan sulfate compounds with the structures shown in Formula I or II. The fucoidan sulfate compounds of the present invention have high anticoagulant activity, and compounds 29, 30, 37, and 58 have relatively strong anticoagulant activity.

[0062] In the present invention, the glycosidic bond of the fucoidan sulfate compound with the structure shown in Formula I is α-1,3 linkage: Series 1: Sulfated at the 2-position (n = 0 to 10), including compounds 1 to 11; Series 2: Sulfated at the 4-position (n = 0 to 10), including compounds 12 to 22; Series 3: Sulfated at the 2,4-positions (n = 0 to 10), including compounds 23 to 30 and Formulas E, F, and G.

[0063] In the present invention, the glycosidic bond of the fucoidan sulfate compound with the structure shown in Formula II is an alternating α-1,3 and α-1,4 linkage. Series 4: Sulfated at the 2,3-positions (n = 0 to 5), including compounds 31 to 41; Series 5: Sulfated at the 3,4-positions (n = 0 to 5), including compounds 42 to 52; Series 6: Sulfated at the 2,3,4-positions (n = 0 to 2), including compounds 53 to 58.

[0064] The present invention also provides a preparation method for the fucoidan sulfate compounds described in the above technical solutions.

[0065] Synthesis of Type I series fucoidan sulfate (Formula I)

[0066] The present invention designs two monosaccharide building blocks 1-4 and 1-5 of fucose. The 2-position of the building block is protected with a benzyl group, and the 4-position is protected with a benzoyl group. The 3-position of the monosaccharide building block 1-5 is protected with a temporary protecting group acetyl, and after removal, the monosaccharide building block 1-4 can be obtained. The specific preparation processes of the monosaccharide building blocks 1-4 and 1-5 are as Figure 1 shown.

[0067] The present invention replaces the 3-position protecting group of the donor with an electron-donating group TBS that can improve the reaction activity to obtain the monosaccharide building block 1-6 (see Figure 2 ). After replacement, there is no problem of aglycone regeneration in the glycosylation reaction. Subsequently, the monosaccharide acceptor 1-8 is obtained through two simple deprotection reactions (see Figure 3 ).

[0068] After obtaining the fucose monosaccharide building blocks (1-4, 1-6, 1-8), a one-pot synthesis strategy based on the pre-activation of the glycosyl donor is used to assemble the fucosooligosaccharide fragments. In the one-pot glycosylation reaction, p-TolSOTf in-situ generated from p-TolSCl / AgOTf is used as a promoter. Since strong acidic TfOH is generated during the reaction, to avoid the detachment of the acid-sensitive TBS silicon protecting group in the donor, an excessive amount of the bulky base TTBP is pre-added to the reaction system as an acid-binding agent to prevent the donor from being damaged.

[0069] The present invention synthesizes a tetrasaccharide donor of fucoidan in one pot. Finally, it is determined that when the equivalent ratio of the four components is 1.0 / 0.95 / 0.85 / 0.85 (1-6 / 1-4 / 1-4 / 1-4), the acceptor can be completely consumed in the first three steps of the glycosylation reaction. At the same time, appropriately increasing the equivalent of the acceptor in the last step can consume as much of the trisaccharide intermediate as possible, thereby improving the separation yield. Finally, the yield of the tetrasaccharide donor 1-15 can be increased to 73% (the reaction yield before optimization was 52%), and the reaction scale can reach the gram level (see Figure 4 ).

[0070] To obtain fucoidan with more sugar components, the present invention synthesizes a fucosylated tetrasaccharide fragment 2-3 with a closed end in one pot, and the yield is 72% (see Figure 5 ).

[0071] The present invention performs a one-step coupling of the monosaccharides 1-4 and 1-6 to obtain the disaccharide 1-13 with a yield of 91%; performs a one-step coupling of the monosaccharides 1-6 and 1-8 to obtain the disaccharide 2-1 with a yield of 93%. Subsequently, the temporary protecting group TBS of the disaccharides 1-13, 2-1, and the tetrasaccharide 1-15 is removed to obtain the corresponding disaccharide acceptor 1-14, 3-1, and tetrasaccharide acceptor 1-16 (see Figure 6 ).

[0072] After completing the one-pot four-component synthesis using monosaccharides as building blocks, the present invention conducts a one-pot glycosylation reaction using disaccharides as building blocks.

[0073] The present invention explores the reaction conditions for the one-pot four-component synthesis of oligosaccharides using disaccharides as building blocks. AgOTf is dissolved in toluene and slowly added in advance at low temperature to ensure the full dispersion of silver ions in the reaction system. Then p-TolSCl is added to ensure that p-TolSCl reacts immediately with AgOTf to generate p-TolSOTf as soon as it is added to the reaction system, activating the donor to obtain an oxonium ion active intermediate, avoiding the formation of chlorinated sugar by-products, and achieving the rapid coupling of 4 disaccharide fragments to obtain the target octasaccharide 2-7 (see Figure 7 ). It should be noted that when conducting a multi-component one-pot reaction using oligosaccharides as building blocks, the system is relatively complex. When using the PE / EA developing system for TLC monitoring to determine whether the donor is fully activated or the acceptor is completely depleted, the R f values of the newly formed sugar and the acceptor used are relatively close, which is prone to misjudgment. Therefore, the Tol / MeCN system is used for monitoring.

[0074] The construction is carried out using the "4 + 4 + 2 + 2" four-component one-pot synthesis method. As Figure 8 shown, the target dodecasaccharide 2-11 is finally obtained in a yield of 60%.

[0075] Through the above experiments, it is proved that the strategy of conducting one-pot glycosylation reactions using monosaccharides, disaccharides, and tetrasaccharides as core sugar building blocks is feasible. Based on this, in the synthesis of the remaining fucoidan fragments, the present invention flexibly combines the above three sugar building blocks to improve the synthesis efficiency of glycosylation. After synthesizing fucoidan fragments of different lengths, the temporary TBS protecting groups on the sugar chain are removed to obtain fucoidans 3-1 to 3-11. The process of one-pot synthesizing the target trisaccharides 2-2 and 3-2 is as Figure 9 shown, and the process of one-pot synthesizing the target tetrasaccharides 2-3 and 3-3 is as Figure 10As shown, similarly, using compounds 1-13, 1-4, and 3-1 as synthetic building blocks, the target pentasaccharide was synthesized using a one-pot three-component "2+1+2" glycosylation strategy; using compounds 1-13, 1-14, and 3-1 as synthetic building blocks, the target hexasaccharide was synthesized using a one-pot three-component "2+2+2" glycosylation strategy; using compounds 1-15, 1-4, and 3-1 as synthetic building blocks, the target heptasaccharide was synthesized using a one-pot three-component "4+1+2" glycosylation strategy; using compounds 1-13, 1-14, and 3-1 as synthetic building blocks, the target octasaccharide was synthesized using a one-pot four-component "2+2+2+2" glycosylation strategy; using compounds 1-15, 1-14, and 1-8 as synthetic building blocks, the target nonasaccharide was synthesized using a one-pot four-component "4+2+2+1" glycosylation strategy; using compounds 1-15, 1-14, and 3-1 as synthetic building blocks, the target decasaccharide was synthesized using a one-pot four-component "4+2+2+2" glycosylation strategy; using compounds 1-15, 1-16, 1-14, and 1-8 as synthetic building blocks, the target undecasaccharide was synthesized using a one-pot four-component "4+4+2+1" glycosylation strategy; using compounds 1-15, 1-16, 1-14, and 3-1 as synthetic building blocks, the target dodecasaccharide was synthesized using a one-pot four-component "4+4+2+2" glycosylation strategy.

[0076] After obtaining a series of fucan fragments with different sugar chain lengths, the protecting groups on the sugar ring were selectively removed, including the following steps:

[0077] Only the benzyl group was removed at the 2-position: For fucans with 2-8 sugar chains, Pd(OH)2 / C of equal mass was used as the catalyst, and the solvent was a mixed solution of MeOH / EA with a volume ratio of 1:1. Stir at room temperature for 6 h under a H2 atmosphere with a pressure of 0.4 MPa; for fucans with 9-12 sugar chains, Pd(OH)2 / C of equal weight was used as the catalyst, and the solvent was a mixed solvent of MeOH / EA with a volume ratio of 1.5:1. Stir at room temperature for 6 h under a H2 atmosphere with a pressure of 0.4 MPa;

[0078] Only the benzoyl group was removed at the 4-position: Using a mixed solution of THF / MeOH with a volume ratio of 3:1 as the solvent, after adjusting the pH value to 10 with solid KOH powder, react at room temperature for 4 h;

[0079] After removing the benzyl group at the 2-position, the benzoyl group was removed at the 4-position: Using anhydrous methanol as the solvent, after adjusting the pH value to 10 with MeONa / MeOH, heat and react at 50 °C for 4 h.

[0080] The present invention carried out sulfation of Type I series fucans, including the following steps:

[0081] 1) Sulfation modification was carried out at the 2-position or 4-position on the fucan sugar ring

[0082] Using trimethylamine trioxide (SO3·NMe3) as the sulfating agent, anhydrous DMF as the reaction solvent, and with 15 equivalents of SO3·NMe3 corresponding to each naked hydroxyl group on the sugar ring, reacting at 50 °C for 48 h can obtain a fucoidan product with hydroxyl groups completely modified by sulfate groups. After obtaining the products with sulfation modification at the 2- or 4-position on the fucoidan sugar ring, further remove the excess protecting groups on the sugar ring to obtain the final product, fucoidan sulfate.

[0083] Sulfation at the 2-position: After the crude product obtained by sulfation is separated by a gel column, it is reacted with 3M NaOH solution at room temperature for 4 h. While removing the benzoyl group, replace NMe3 salted with sulfate with a more stable sodium salt to obtain fucoidan sulfate compounds 1 - 11 with sulfation at the 2-position. Figure 11 Schematic diagram for obtaining fucoidan sulfate compounds 1 - 11.

[0084] Sulfation at the 4-position: After the crude product obtained by sulfation is separated by a gel column, use 10 times the weight of Pd(OH)2 / C as the catalyst, with the solvent being a mixed solution of MeOH / H2O with a volume ratio of 1:1. Stir at room temperature for 16 h under a H2 atmosphere with a pressure of 0.4 MPa, and then treat with 3M NaOH solution for 30 min to replace NMe3 salted with sulfate with a more stable sodium salt to obtain fucoidan sulfate compounds 12 - 22 with sulfation at the 4-position. Figure 12 Schematic diagram for obtaining fucoidan sulfate compounds 12 - 22.

[0085] Sulfation modification at both the 2- and 4-positions on the fucoidan sugar ring: When using pyridine trioxide (SO3·Pyr) with 15 equivalents corresponding to each naked hydroxyl group on the sugar ring as the sulfating agent, the obtained product is closer to the target product compared to other sulfating agents, that is, the proportion of hydroxyl groups sulfated is higher. React at room temperature for 14 h, while appropriately reducing the volume of the reaction solvent, and after treatment with 3M NaOH solution, successfully obtain fully sulfated fucoidan sulfate compounds 23 - 30 and formulas E, F, and G. Figure 13 Schematic diagram for obtaining fucoidan sulfate compounds 23 - 30 and formulas E, F, and G.

[0086] Synthesis of type II series fucoidan sulfate (formula II)

[0087] Based on the monosaccharide building blocks of the 1,3-linkage series, two building blocks 1 - 10 and 1 - 12 were designed, as Figure 14 shown.

[0088] After obtaining the monosaccharide building blocks 1-10 and 1-12, the oligosaccharide fragments were assembled using a one-pot reaction strategy. Since the PMB group is more sensitive to acidic environments, the amount of the acid scavenger TTBP was increased in the reaction, from 1.2 eq of TTBP corresponding to each portion of the p-TolSCl / AgOTf promoter to 1.3 eq. The processes of obtaining fucotetraose 1-19 and 10-3 are shown respectively as Figure 15 and 16 shown.

[0089] The disaccharide 1-17 was obtained by one-step coupling of the monosaccharides 1-8 and 1-10; the disaccharide 10-1 was obtained by one-step coupling of the monosaccharides 1-8 and 1-12. Subsequently, when using HF·Pyr to remove the TBS protecting group, since the PMB protecting group contained in Type II fucoidan is not very stable and sensitive to acidic environments, too high a reaction temperature or too long a reaction time would both cause the PMB to fall off, resulting in a significant reduction in yield. Based on this, the reaction temperature was lowered, and the glycosyl acceptors 1-18, 1-20, 11-1, and 11-3 were successfully obtained (see Figure 17 ).

[0090] After obtaining the glycosyl donors and acceptors, the above sugar building blocks were flexibly combined to synthesize fucosylan fragments with different lengths and 1,3-, 1,4-alternating linkages. Finally, the temporary TBS protecting group on the sugar chain was removed to obtain fucosylated oligosaccharides. Using the compound 1-6, 1-4, and 1-12 as synthetic building blocks and the one-pot three-component "1+1+1" glycosylation strategy, the target trisaccharide 10-2 was synthesized, as shown in Figure 18 , and using the compound 1-6, 1-10, 1-4, and 1-12 as synthetic building blocks and the one-pot four-component "1+1+1+1" glycosylation strategy, the target tetrasaccharide 10-3 was synthesized, as shown in Figure 19, using compounds 1-6, 1-18, and 11-1 as synthetic building blocks, the target pentasaccharide was synthesized using a one-pot three-component "1+2+2" glycosylation strategy. Using compounds 1-17, 1-18, and 11-1 as synthetic building blocks, the target hexasaccharide was synthesized using a one-pot three-component "2+2+2" glycosylation strategy. Using compounds 1-6, 1-18, and 11-1 as synthetic building blocks, the target heptasaccharide was synthesized using a one-pot four-component "1+2+2+2" glycosylation strategy. Using compounds 1-19, 1-18, and 11-1 as synthetic building blocks, the target octasaccharide was synthesized using a one-pot three-component "4+2+2" glycosylation strategy. Using compounds 1-6, 1-18, and 11-3 as synthetic building blocks, the target nonasaccharide was synthesized using a one-pot four-component "1+2+2+4" glycosylation strategy. Using compounds 1-19, 1-18, and 11-3 as synthetic building blocks, the target decasaccharide was synthesized using a one-pot three-component "4+2+4" glycosylation strategy. Using compounds 1-6, 1-20, 1-18, and 11-3 as synthetic building blocks, the target undecasaccharide was synthesized using a one-pot four-component "1+4+2+4" glycosylation strategy. Using compounds 1-19, 1-20, and 11-3 as synthetic building blocks, the target dodecasaccharide was synthesized using a one-pot three-component "4+4+4" glycosylation strategy.

[0091] After obtaining a series of fucoidan fragments with different sugar chain lengths, the protecting groups on the sugar ring were selectively removed. Different from Type I series fucoidan, there are three sites on the sugar ring of Type II series fucoidan that can be sulfated, namely the 2nd, 3rd, and 4th positions.

[0092] Removing the PMB protecting group at the 3rd position on the sugar ring

[0093] First, the PMB protecting group on the sugar ring was removed. Using a mixed solution with a DCM / TfOH volume ratio of 10:1 as the solvent, the reaction was carried out at room temperature for 30 min to obtain compounds 12-1 to 12-11 (see Figure 20 ).

[0094] On the fucoidan from which the PMB protecting group has been removed, the benzyl group at the 2nd position on the sugar ring was further removed. For fucoidan with 2 to 8 sugar chains, Pd(OH)2 / C with an equal weight was used as the catalyst, and a mixed solution with a MeOH:EA volume ratio of 1:1 was used as the solvent. The reaction was stirred at room temperature for 6 h under a H2 atmosphere with a pressure of 0.4 MPa, and the benzyl group at the 2nd position on the sugar ring could be completely removed. For fucoidan with 9 to 12 sugar chains, a mixed solution with a MeOH:EA volume ratio of 1.5:1 was used as the solvent instead, and finally compounds 13-1 to 13-11 were obtained, see Figures 21 - 22 .

[0095] On the fucan from which the PMB protecting group has been removed, the benzoyl group at the 4-position on the sugar ring is further removed. The reaction conditions are as follows: using a mixed solution with a volume ratio of THF / MeOH = 3:1 as the solvent, adjusting the pH value to 10 with solid KOH powder, and then reacting at room temperature for 4 h. Finally, compounds 14-1 to 14-11 are obtained (see Figure 23 ).

[0096] On the fucan from which the PMB protecting group has been removed, the benzyl group at the 2-position and the benzoyl group at the 4-position on the sugar ring are further removed. The reaction conditions are as follows: using anhydrous methanol as the solvent, adjusting the pH value to 10 with MeONa / MeOH, and then heating and reacting at 50 °C for 4 h. Finally, compounds 15-1 to 15-6 are obtained (see Figure 24 ).

[0097] The present invention performs sulfation of Type II series fucans, including the following steps:

[0098] Using sulfur trioxide pyridine (SO3·Pyr) as the sulfating agent and anhydrous DMF as the reaction solvent, when each naked hydroxyl group on the sugar ring corresponds to 10 equivalents of (SO3·Pyr), reacting at room temperature for 7 h can obtain a fucan intermediate in which the hydroxyl groups are completely modified by sulfate groups.

[0099] Sulfation modification at the 2,3-position: After obtaining the fucan intermediate, further remove the excess protecting groups on the sugar ring through post-treatment. After the obtained crude product is separated by a gel column, it is reacted with 3M NaOH solution at room temperature for 4 h. While removing the benzoyl group, the pyridinium salt formed with the sulfate group is replaced with a more stable sodium salt. Finally, fucan sulfate compounds 31 to 41 with sulfation at the 2,3-position are obtained.

[0100] Sulfation modification at the 3,4-position: After obtaining the fucan intermediate, further remove the excess protecting groups on the sugar ring through post-treatment. After the obtained crude product is separated by a gel column, 10 times the weight of Pd(OH)2 / C is used as the catalyst, and the solvent is a mixed solution with a volume ratio of MeOH / H2O = 1:1. Stir at room temperature for 16 h under a H2 atmosphere with a pressure of 0.4 MPa, and then treat with 3M NaOH solution for 30 min to replace the pyridinium salt formed with the sulfate group with a more stable sodium salt. Finally, fucan sulfate compounds 42 to 52 with sulfation at the 3,4-position are obtained.

[0101] 2,3,4-Sulfation modification: Using pyridine sulfur trioxide (SO3·Pyr) as the sulfating reagent and anhydrous DMF as the reaction solvent, with 10 equivalents of SO3·Pyr corresponding to each naked hydroxyl group on the sugar ring, reacting at room temperature for 14 h, and then treating with 3M NaOH solution for 30 min, sulfated fucoidan compounds 53-58 at the 2,3,4-positions can be obtained.

[0102] The present invention also provides the use of the fucoidan compounds described in the above technical solution in the preparation of anticoagulant active drugs.

[0103] In the present invention, the dosage form of the anticoagulant active drug preferably includes pharmaceutically acceptable dosage forms such as tablets, injections, capsules, granules, pills, powders, oral liquids, sustained-release preparations, controlled-release preparations or nano-preparations.

[0104] In the present invention, the anticoagulant active drug preferably contains an effective dose of the fucoidan compounds, pharmaceutically acceptable salts and pharmaceutically acceptable carriers, excipients, excipients and diluents.

[0105] Next, the technical solution in the present invention will be clearly and completely described in conjunction with the examples in the present invention. Obviously, the described examples are only a part of the examples of the present invention, rather than all the examples. Based on the examples in the present invention, all other examples obtained by those of ordinary skill in the art without creative efforts belong to the protection scope of the present invention.

[0106] All chemical reagents used in the experimental process are of reagent grade and do not need to be purified again before use; anhydrous dichloromethane (DCM), acetonitrile (MeCN), and toluene (Tol) are all obtained by redistilling commercially available solvents (analytical grade) with calcium hydride. The mixed liquid system after the reaction is monitored by TLC (Thin-Layer Chromatography, manufacturer: E.Merck F254), that is, after the eluent develops the plate with the reaction solution, the spots are first observed under ultraviolet light, then stained with a color-developing agent (cerium molybdate), and then baked to develop the color; the silica gel used in the column chromatography operation is 200-300 mesh silica gel (Qingdao Ocean Chemical Co., Ltd.) or 300-400 mesh silica gel (Merck); the nuclear magnetic data is recorded using a Bruke AVANCE III-600 nuclear magnetic resonance spectrometer. 1 The chemical shift of 1H NMR is based on tetramethylsilane as the internal standard or the chemical shift of the residual hydrogen signal of the deuterated solvent as the standard (when the deuterated solvent itself does not contain tetramethylsilane). 13The chemical shifts of 13C NMR were referenced to the chemical shifts of the deuterated solvents; high-resolution mass spectrometry was recorded using a Thermo Scientific LTQ Orbitrap Discovery or a Waters Xevo G2 QTof mass spectrometer.

[0107] Example 1

[0108] To prepare the compound of the structure shown in Formula 29, the following steps are included:

[0109] Compound 1-4

[0110]

[0111] L-Fucose (5.00 g, 30.36 mmol) was added to a 100 mL round-bottom flask containing acetic anhydride (20 mL), and the round-bottom flask was stirred in an ice bath. After the temperature of the reaction solution dropped from room temperature to 0 °C, perchloric acid (0.2 mL, 3.00 mmol) was slowly added dropwise to the round-bottom flask. After waiting for about 5 min, the ice bath was removed, and the reaction solution was continuously stirred at room temperature. After 1 h, TLC was used for monitoring. After it was shown that the reaction was complete, triethylamine (1 mL) was added dropwise to the reaction solution to quench the reaction, ethyl acetate (20 mL) was added to dilute the reaction solution, and then the reaction solution was extracted three times with water (60 mL × 3), saturated sodium bicarbonate solution (60 mL × 3), and saturated brine (60 mL × 3) in sequence. The aqueous phase was discarded, the organic phase was retained, and it was dried over anhydrous sodium sulfate. After filtering off the desiccant, the filtrate was concentrated to obtain the crude product 1-1.

[0112] Compound 1-1 and p-toluenethiol (4.54 g, 36.43 mmol) were added to a 100 mL round-bottom flask containing anhydrous dichloromethane (20 mL), and the round-bottom flask was stirred in an ice bath. After the temperature of the reaction solution dropped from room temperature to 0 °C, boron trifluoride etherate solution (15 mL) was slowly added to the round-bottom flask. After waiting for about 10 min, the ice bath was removed, and the reaction solution was continuously stirred at room temperature. After 1 h, TLC was used for monitoring. After it was shown that the reaction was complete, saturated sodium bicarbonate solution (20 mL) was added to the reaction solution to quench the reaction. Then, the reaction solution was extracted with dichloromethane (50 mL × 3), and the organic phases were combined. The combined organic phase was washed three times with water (60 mL × 3), saturated sodium bicarbonate solution (60 mL × 3), and saturated brine (60 mL × 3) in sequence. The aqueous phase was discarded, the organic phase was retained, and it was dried over anhydrous sodium sulfate. After filtering off the desiccant, the filtrate was concentrated to obtain the crude product 1-2.

[0113] Compound 1-2 was added to a 100 mL round-bottom flask containing anhydrous methanol (20 mL). After the compound was completely dissolved, the round-bottom flask was placed in an oil bath with the temperature set at 50 °C. A solution of sodium methoxide / methanol (2 mL) was added dropwise to the round-bottom flask, and the reaction was stirred thoroughly. After 1 h, TLC was used for monitoring, indicating that the reaction was complete. The heating was stopped and the oil bath was removed. After the reaction solution was cooled to room temperature, cation exchange resin was added to the reaction solution until the pH of the reaction solution was neutral. Then, the adjusted reaction solution was filtered through diatomaceous earth to remove the resin, and the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (petroleum ether / ethyl acetate, volume ratio 2:1) to obtain white solid 1-3 (6.80 g, 80% for 3 steps).

[0114] Compound 1-3 (3.70 g, 13.7 mmol) was added to a 100 mL round-bottom flask containing anhydrous acetonitrile (40 mL). After the compound was completely dissolved, PhC(OMe)3 (7.10 mL, 41.1 mmol) and camphorsulfonic acid (330 mg, 1.40 mmol) were added, and the reaction was stirred at room temperature. After 3 h, TLC was used for monitoring. After indicating that the reaction was complete, triethylamine (5 mL) was added dropwise to the reaction solution to quench the reaction, and the color of the reaction solution changed from dark red to light yellow. Subsequently, the reaction solution was concentrated, and anhydrous DMF (30 mL) was added to the round-bottom flask. The round-bottom flask was placed in an ice bath and stirred. After the temperature of the reaction solution dropped from room temperature to 0 °C, BnBr (4.70 mL, 27.4 mmol) was first added to the round-bottom flask, and then NaH (1.10 g, 27.4 mmol) was added slowly in portions. After waiting for about 30 min, the ice bath was removed, and the reaction solution was continuously stirred at room temperature. After 3 h, TLC was used for monitoring again. After indicating that the reaction was complete, the reaction solution was diluted with dichloromethane (20 mL), and the round-bottom flask was placed in an ice bath. 1N HCl (10 mL) was slowly added dropwise until no more bubbles were generated in the flask. Subsequently, the ice bath was removed, and the reaction was continuously stirred at room temperature. After 1 h, TLC was used for monitoring again, indicating that the reaction was complete. Then, the reaction solution was extracted with dichloromethane (50 mL × 3), and the organic phases were combined. The organic phase was washed three times successively with water (60 mL × 3), saturated sodium bicarbonate solution (60 mL × 3), and saturated brine (60 mL × 3). The aqueous phases were discarded, and the organic phase was retained and dried over anhydrous sodium sulfate. After filtering off the desiccant, the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (petroleum ether / ethyl acetate, volume ratio 6:1) to obtain colorless syrup 1-4 (5.08 g, 80% for 3 steps). R f = 0.40 (PE / EA = 3:1, volume ratio); 11H NMR (400 MHz, CDCl3) δ 8.07 - 8.02 (m, 2H), 7.60 (m, 3H), 7.47 (m, 2H), 7.43 - 7.39 (m, 2H), 7.37 - 7.27 (m, 3H), 7.18 (d, J = 7.9 Hz, 2H), 5.38 - 5.32 (m, 1H), 4.93 (d, J = 10.7 Hz, 1H), 4.69 (d, J = 10.7 Hz, 1H), 4.61 (d, J = 9.6 Hz, 1H), 3.89 (dt, J = 9.3, 3.2 Hz, 1H), 3.80 - 3.73 (m, 1H), 3.64 (t, J = 9.4 Hz, 1H), 2.70 (d, J = 3.7 Hz, 1H), 2.40 (s, 3H), 1.30 - 1.26 (m, 3H); 13 13C NMR (101 MHz, CDCl3) δ 166.77, 138.23, 137.86, 133.39, 133.10, 130.12, 129.77, 129.64, 129.45, 128.58, 128.45, 128.29, 128.05, 86.95, 77.48, 75.32, 74.22, 73.68, 73.51, 21.33, 16.91; HRMS (ESI) Calcd for C 27 H 28 O5SNa [M + Na] + : 487.1555, found: 487.1549.

[0115] Compounds 1 - 6

[0116]

[0117] Compounds 1 - 4 (2.80 g, 6.00 mmol) were added to a 100 mL round - bottom flask containing anhydrous DMF (40 mL). After the compounds were completely dissolved, imidazole (1.02 g, 15.0 mmol) and tert - butyldimethylchlorosilane (TBSCl) (2.27 g, 15.0 mmol) were added in sequence. Then the round - bottom flask was placed in an oil bath at 70 °C and stirred vigorously for reaction. After 4 h, TLC was used to monitor and showed that the reaction was complete. The heating was stopped and the oil bath was removed. After the reaction solution was cooled to room temperature, ethyl acetate (20 mL) was added to dilute the reaction solution, and then it was extracted three times with water (60 mL × 3), saturated sodium bicarbonate solution (60 mL × 3), and saturated brine (60 mL × 3) in sequence. The aqueous phase was discarded, and the organic phase was retained and dried over anhydrous sodium sulfate. After filtering off the desiccant, the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (petroleum ether / ethyl acetate, 35:1) to obtain white solid 1 - 6 (3.31 g, 95%). Rf = 0.70 (PE / EA volume ratio = 6:1); 1 HNMR (400 MHz, CDCl3) δ 8.05 - 8.00 (m, 2H), 7.63 - 7.54 (m, 3H), 7.49 - 7.42 (m, 4H), 7.38 - 7.27 (m, 3H), 7.17 - 7.12 (m, 2H), 5.34 (dd, J = 3.5, 1.0 Hz, 1H), 4.80 (d, J = 10.3 Hz, 1H), 4.69 (d, J = 10.3 Hz, 1H), 4.62 (d, J = 9.7 Hz, 1H), 3.91 (dd, J = 8.8, 3.4 Hz, 1H), 3.85 - 3.79 (m, 1H), 3.63 (dd, J = 9.7, 8.8 Hz, 1H), 2.39 (s, 3H), 1.28 (d, J = 6.4 Hz, 3H), 0.76 (s, 9H), 0.11 (s, 3H), 0.03 (s, 3H); 13 C NMR (101 MHz, CDCl3) δ 166.28, 138.47, 137.70, 133.12, 133.02, 130.19, 130.03, 129.87, 129.77, 128.42, 128.38, 128.11, 127.72, 87.47, 77.74, 75.54, 75.11, 74.29, 73.51, 25.77, 21.39, 17.84, 17.04, -4.53, -4.71; HRMS (ESI) Calcd for C 33 H 46 NO5SSi [M+NH4] + : 596.2866, found: 596.2878.

[0118] Compound 1 - 7

[0119]

[0120] Take a 100 mL two - neck reaction flask, add a magnetic stir bar, evacuate the flask together with the stir bar to vacuum, then bake the reaction flask with a high - temperature torch until no water mist appears on its inner wall. After the flask cools to room temperature, add the pre - baked Molecular sieve (2.00 g) was evacuated, and then the molecular sieve was baked and activated with a high-temperature spray gun. After the molecular sieve showed a phenomenon of being dispersed and sticking to the bottle wall, the baking was stopped. After waiting for the reaction flask to cool to room temperature again, protective gas argon was introduced. Subsequently, anhydrous dichloromethane (20 mL) was added. Then, glycosyl donor 1-6 (0.579 g, 1.00 mmol) and TTBP (0.300 g, 1.20 mmol) were dissolved together in anhydrous dichloromethane (20 mL) and added to the reaction flask. After stirring at room temperature for 20 min, the reaction temperature was lowered to -78 °C. After 5 min, AgOTf (0.514 g, 2.00 mmol) pre-dissolved in anhydrous toluene (8 mL) was slowly added dropwise. Subsequently, a stoichiometric amount of p-TolSCl (133.3 μL, 1.00 mmol) was added, and the mixture was stirred for 5 min. Then TLC was used for monitoring, and it showed that glycosyl donor 1-6 completely disappeared, indicating that the pre-activation reaction was complete. Immediately, anhydrous methanol (1 mL) was slowly added dropwise to the reaction solution. After the temperature slowly rose to room temperature, TLC was continued to be used for monitoring, and it showed that the reaction ended. Finally, triethylamine (0.5 mL) was added to quench the reaction. The quenched reaction solution was filtered through diatomaceous earth to remove the molecular sieve, and the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (petroleum ether / ethyl acetate, 30:1) to obtain colorless syrup 1-7 (0.448 g, 92%). R f = 0.45 (volume ratio of PE / EA = 2:1); 1 1H NMR (400 MHz, CDCl3) δ 8.11 (d, J = 7.6 Hz, 2H), 7.57 (m, 1H), 7.46 (m, 2H), 7.33 (m, 5H), 5.32 (d, J = 3.7 Hz, 1H), 4.93 (d, J = 10.8 Hz, 1H), 4.71 (d, J = 10.8 Hz, 1H), 4.35 (d, J = 7.8 Hz, 1H), 3.86 (dd, J = 9.3, 3.7 Hz, 1H), 3.79 (q, J = 6.4 Hz, 1H), 3.61 (s, 3H), 3.57 (m, 1H), 1.27 (d, J = 6.3 Hz, 3H), 0.77 (s, 9H), 0.10 (s, 3H), 0.04 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 166.45, 138.82, 133.02, 130.25, 130.03, 128.39, 128.28, 128.13, 127.52, 105.21, 79.94, 75.14, 74.07, 73.06, 69.35, 57.44, 25.71, 17.94, 16.56, -4.65, -4.74; HRMS (ESI) Calcd for C 27 H 42NO6Si[M+NH4] + : 504.2781, found: 504.2782.

[0121] Compounds 1 - 8

[0122]

[0123] Compounds 1 - 7 (0.487 g, 1.00 mmol) were added to a 50 mL plastic reaction tube and dissolved in anhydrous tetrahydrofuran (10 mL). Subsequently, the reaction tube was placed in an ice bath and stirred. After the temperature dropped from room temperature to 0 °C, 70 wt% HF - Pyridine (2 mL) was slowly added dropwise. After waiting for about 5 min, the ice bath was removed, and the plastic reaction tube was placed in an oil bath pot set at 50 °C and stirred thoroughly for the reaction. After 4 h, TLC was used to monitor, indicating that the reaction was complete. The heating was stopped and the oil bath pot was removed. After the reaction environment temperature cooled to room temperature, ethyl acetate (10 mL) was added for dilution. Subsequently, the reaction tube was placed in an ice bath and stirred, and triethylamine (5 mL) was slowly added dropwise to quench the reaction. Then the reaction solution was concentrated to obtain the crude product. The crude product was further separated by column chromatography (petroleum ether / ethyl acetate, volume ratio 8:1) to obtain colorless syrup - like substance 1 - 8 (0.354 g, 95%). R f = 0.30 (PE / EA volume ratio = 2:1); 1 1H NMR (400 MHz, CDCl3) δ 8.12 - 8.04 (m, 2H), 7.61 - 7.54 (m, 1H), 7.44 (m, 2H), 7.39 - 7.26 (m, 5H), 5.41 (dd, J = 3.6, 1.1 Hz, 1H), 4.98 (d, J = 11.3 Hz, 1H), 4.69 (d, J = 11.3 Hz, 1H), 4.35 (d, J = 7.7 Hz, 1H), 3.86 (dt, J = 9.7, 3.1 Hz, 1H), 3.80 (qd, J = 6.4, 1.1 Hz, 1H), 3.64 - 3.56 (m, 4H), 2.49 (d, J = 3.0 Hz, 1H), 1.26 (d, J = 6.3 Hz, 3H); 13 13C NMR (101 MHz, CDCl3) δ 166.69, 138.44, 133.30, 130.13, 129.72, 128.59, 128.46, 128.23, 127.96, 104.86, 79.23, 74.78, 72.89, 72.21, 69.66, 57.33, 16.49; HRMS (ESI) Calcd for C 21 H 28 NO6[M+NH4] +: 390.1917, found: 390.1911.

[0124] Compounds 1 - 13

[0125]

[0126] Take a 100 mL two - necked reaction flask, add a magnetic stir bar, and evacuate the flask together with the stir bar to vacuum. Then bake the reaction flask with a high - temperature blowtorch until no water mist appears on the inner wall of the flask. After the flask cools to room temperature, add the pre - baked molecular sieve (2.00 g). Evacuate the flask to vacuum and then bake and activate the molecular sieve with a high - temperature blowtorch. Stop baking when the molecular sieve shows a phenomenon of dispersing and sticking to the inner wall of the flask. Wait again for the reaction flask to cool to room temperature, then introduce the protective gas argon. Subsequently, add anhydrous dichloromethane (20 mL). Then dissolve the glycosyl donor 1 - 6 (0.579 g, 1.00 mmol) and TTBP (0.300 g, 1.20 mmol) together in anhydrous dichloromethane (20 mL) and add them to the reaction flask. Stir at room temperature for 20 min, then lower the reaction temperature to - 78 °C. After 5 min, slowly add dropwise AgOTf (0.514 g, 2.00 mmol) pre - dissolved in anhydrous toluene (8 mL), and then add a stoichiometric amount of p - TolSCl (133.3 μL, 1.00 mmol), and stir for 5 min. Then monitor the reaction using TLC. When it shows that the glycosyl donor 1 - 6 has completely disappeared, indicating that the pre - activation reaction is complete, immediately slowly add dropwise the glycosyl acceptor 1 - 4 (0.465 g, 1.00 mmol) pre - dissolved in anhydrous dichloromethane (2 mL). After the reaction temperature slowly rises to room temperature, monitor the reaction using TLC. When it shows that the reaction is over, then add triethylamine (0.5 mL) to quench the reaction. Filter the quenched reaction solution through diatomaceous earth to remove the molecular sieve, concentrate the filtrate to obtain the crude product. The crude product is further separated by column chromatography (petroleum ether / ethyl acetate, volume ratio 8:1) to obtain white solid 1 - 13 (0.827 g, 90%). R f = 0.40 (PE / EA volume ratio = 2:1); 11H NMR (400 MHz, CDCl3) δ 7.97 (m, 4H), 7.62 - 7.53 (m, 4H), 7.47 - 7.32 (m, 9H), 7.21 - 7.11 (m, 7H), 5.67 (dd, J = 3.1, 0.9 Hz, 1H), 5.32 (d, J = 3.4 Hz, 1H), 5.14 (dd, J = 3.5, 1.3 Hz, 1H), 5.02 (d, J = 10.4 Hz, 1H), 4.78 (d, J = 10.4 Hz, 1H), 4.68 (d, J = 9.6 Hz, 1H), 4.51 (d, J = 12.0 Hz, 1H), 4.38 (d, J = 11.9 Hz, 1H), 4.25 - 4.17 (m, 2H), 4.03 (dd, J = 9.4, 3.1 Hz, 1H), 3.87 (t, J = 9.5 Hz, 1H), 3.82 - 3.74 (m, 2H), 2.42 (s, 3H), 1.29 (d, J = 6.4 Hz, 3H), 0.97 (d, J = 6.5 Hz, 3H), 0.70 (s, 9H), 0.01 (s, 3H), -0.06 (s, 3H); 13 13C NMR (101 MHz, CDCl3) δ 166.43, 166.37, 138.34, 138.27, 137.89, 133.23, 133.22, 132.91, 130.49, 130.18, 129.88, 129.85, 129.81, 129.73, 128.64, 128.44, 128.40, 128.07, 128.03, 127.85, 127.79, 127.25, 93.86, 87.69, 76.51, 76.06, 75.55, 75.28, 75.02, 73.78, 72.19, 69.61, 69.56, 65.45, 25.75, 21.40, 17.90, 17.12, 16.26, -4.65, -4.84; HRMS (ESI) Calcd for C 53 H 66 NO 10 SSi [M + NH4] + : 936.4177, found: 936.4187.

[0127] Compound 1 - 14

[0128]

[0129] Compound 1-13 (0.919 g, 1.00 mmol) was added to a 50 mL plastic reaction tube and dissolved in anhydrous tetrahydrofuran (10 mL). Subsequently, the reaction tube was placed in an ice bath and stirred. After the temperature dropped to 0 °C, 70 wt% HF-Pyridine (2 mL) was slowly added dropwise. After waiting for about 5 min, the ice bath was removed, and the plastic reaction tube was placed in an oil bath pot with a temperature set at 50 °C and stirred thoroughly for the reaction. After 4 h, TLC monitoring showed that the reaction was complete, and the heating was stopped and the oil bath pot was removed. After the reaction environment temperature cooled to room temperature, ethyl acetate (10 mL) was added for dilution. Subsequently, the reaction tube was placed in an ice bath and stirred, and triethylamine (5 mL) was slowly added dropwise to quench the reaction. Then the reaction solution was concentrated to obtain the crude product. The crude product was further separated by column chromatography (petroleum ether / ethyl acetate, volume ratio 5:1) to obtain white solid 1-14 (0.724 g, 90%). R f = 0.30 (PE / EA volume ratio = 2:1); 1 H NMR (600 MHz, CDC l3 ) δ 8.03 - 8.00 (m, 2H), 7.92 - 7.87 (m, 2H), 7.66 - 7.62 (m, 2H), 7.60 (m, 1H), 7.57 - 7.53 (m, 1H), 7.45 - 7.38 (m, 8H), 7.37 - 7.33 (m, 1H), 7.22 - 7.16 (m, 5H), 7.11 (m, 2H), 5.70 (dd, J = 3.2, 1.0 Hz, 1H), 5.43 (d, J = 3.4 Hz, 1H), 5.07 (d, J = 10.4 Hz, 1H), 5.02 (dd, J = 3.4, 1.3 Hz, 1H), 4.68 (d, J = 9.6 Hz, 1H), 4.58 (d, J = 10.4 Hz, 1H), 4.52 (d, J = 12.0 Hz, 1H), 4.35 (d, J = 12.0 Hz, 1H), 4.17 (qd, J = 6.4, 1.4 Hz, 1H), 4.08 (m, 2H), 3.90 - 3.82 (m, 2H), 3.74 (dd, J = 10.1, 3.4 Hz, 1H), 2.43 (s, 3H), 1.98 (d, J = 2.9 Hz, 1H), 1.32 (d, J = 6.4 Hz, 3H), 0.90 (d, J = 6.5 Hz, 3H); 1313C NMR (151 MHz, CDCl3) δ 166.49, 166.45, 138.20, 138.11, 137.59, 133.59, 133.52, 133.11, 130.28, 129.94, 129.89, 129.87, 129.38, 129.10, 128.66, 128.58, 128.52, 128.43, 128.23, 128.15, 127.94, 127.86, 91.95, 87.30, 75.78, 75.73, 75.41, 75.14, 73.62, 73.39, 72.03, 68.69, 67.85, 65.19, 21.42, 17.10, 16.00; HRMS (ESI) Calcd for C 47 H 52 NO 10 S [M+NH4]+: 822.3312, found: 822.3323.

[0130] Compounds 1 - 15

[0131]

[0132] Take a 50 mL two-necked reaction flask, add a magnetic stir bar, evacuate the flask together with the stir bar to vacuum, and then bake the reaction flask with a high-temperature blowtorch until no water mist appears on the inner wall of the flask. After the flask cools to room temperature, add the pre-baked molecular sieve (2.00 g). Evacuate the flask to vacuum, and then bake and activate the molecular sieve with a high-temperature blowtorch. Stop baking when the molecular sieve shows a phenomenon of being dispersed and adhering to the inner wall of the flask. Wait again for the reaction flask to cool to room temperature, then introduce the protective gas argon, and then add anhydrous dichloromethane (10 mL). Then dissolve the glycosyl donor 1 - 6 (57.9 mg, 100.0 μmol) and TTBP (80.0 mg, 320.0 μmol) together in anhydrous dichloromethane (10 mL) and add them to the reaction flask.

[0133] After stirring at room temperature for 20 min, the reaction temperature was lowered to -78 °C. After 5 min, AgOTf (51.4 mg, 200.0 μmol) dissolved in anhydrous toluene (1 mL) was slowly added dropwise. Subsequently, a stoichiometric amount of p-TolSCl (13.4 μL, 100.0 μmol) was added, and the reaction was stirred for 5 min. Then, TLC monitoring was used, showing that the glycosyl donor 1-6 completely disappeared, indicating that the pre-activation reaction was complete. Immediately, the glycosyl acceptor 1-4 (41.8 mg, 90.0 μmol) dissolved in anhydrous dichloromethane (1 mL) was slowly added dropwise to the reaction solution. The temperature condition of -78 °C was maintained, and the reaction was carried out for 20 min. Then, the reaction temperature was slowly raised to room temperature and maintained at room temperature for 20 min. TLC monitoring was used, showing that the glycosyl acceptor 1-4 completely disappeared, and a new spot, namely the disaccharide product, was formed.

[0134] Again, the reaction temperature was lowered to -78 °C. After 5 min, AgOTf (46.3 mg, 180.0 μmol) dissolved in anhydrous toluene (1 mL) was slowly added dropwise. Subsequently, a stoichiometric amount of p-TolSCl (12.0 μL, 90 μmol) was added, and the reaction was stirred for 5 min. Then, TLC monitoring was used, showing that the generated disaccharide product completely disappeared, indicating that the pre-activation reaction was complete. Immediately, the glycosyl acceptor 1-4 (37.2 mg, 80.0 μmol) dissolved in anhydrous dichloromethane (1 mL) was slowly added dropwise to the reaction solution. The temperature condition of -78 °C was maintained, and the reaction was carried out for 20 min. Then, the reaction temperature was slowly raised to room temperature and maintained at room temperature for 20 min. TLC monitoring was used, showing that the glycosyl acceptor 1-4 completely disappeared, and a new spot, namely the trisaccharide product, was formed.

[0135] Again, the reaction temperature was lowered to -78 °C. After 5 min, AgOTf (41.1 mg, 160.0 μmol) dissolved in anhydrous toluene (1 mL) was slowly added dropwise. Subsequently, a stoichiometric amount of p-TolSCl (10.7 μL, 80 μmol) was added, and the reaction was stirred for 5 min. Then, TLC monitoring was used, showing that the generated trisaccharide product completely disappeared, indicating that the pre-activation reaction was complete. Immediately, the glycosyl acceptor 1-4 (37.2 mg, 80.0 μmol) dissolved in anhydrous dichloromethane (1 mL) was slowly added dropwise to the reaction solution. After the reaction temperature was slowly raised to room temperature, TLC monitoring was used, showing that the reaction ended. Then, triethylamine (0.5 mL) was added dropwise to quench the reaction. The quenched reaction solution was filtered through diatomaceous earth to remove the molecular sieve, and the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (toluene / acetonitrile, 8:1) to obtain white solid 1-15 (107.2 mg, 67%). R f = 0.50 (volume ratio of toluene / MeCN = 7:1); 11H NMR (400 MHz, CDCl3) δ 8.06 - 8.01 (m, 2H), 7.94 - 7.85 (m, 6H), 7.64 - 7.58 (m, 3H), 7.57 - 7.48 (m, 3H), 7.39 (m, 14H), 7.25 - 7.06 (m, 18H), 5.67 (d, J = 3.1 Hz, 1H), 5.31 (d, J = 3.5 Hz, 1H), 5.22 (d, J = 3.2 Hz, 1H), 5.14 (dd, J = 6.6, 3.6 Hz, 2H), 5.09 (d, J = 10.2 Hz, 1H), 5.01 (d, J = 3.4 Hz, 1H), 4.96 - 4.93 (m, 1H), 4.70 (d, J = 9.6 Hz, 1H), 4.64 - 4.55 (m, 3H), 4.40 (d, J = 12.1 Hz, 1H), 4.34 (t, J = 11.0 Hz, 2H), 4.27 (dd, J = 10.3, 3.2 Hz, 1H), 4.21 - 4.05 (m, 5H), 4.02 (m, 2H), 3.94 (ddd, J = 17.8, 10.3, 3.4 Hz, 2H), 3.89 - 3.83 (m, 2H), 3.63 (dd, J = 9.7, 3.4 Hz, 1H), 2.41 (s, 3H), 1.31 (d, J = 6.3 Hz, 3H), 0.86 (d, J = 6.5 Hz, 3H), 0.77 (dd, J = 12.8, 6.4 Hz, 6H), 0.63 (s, 9H), -0.13 (s, 3H), -0.22 (s, 3H); 13CNMR (101 MHz, CDCl3) δ 166.61, 166.43, 166.42, 138.49, 138.28, 138.22, 137.96, 137.88, 133.41, 133.21, 133.15, 132.90, 132.84, 130.53, 130.22, 130.18, 130.09, 129.97, 129.92, 129.86, 129.66, 128.65, 128.53, 128.43, 128.35, 128.31, 128.26, 128.17, 128.09, 127.98, 127.96, 127.94, 127.61, 127.52, 127.11, 93.23, 93.16, 93.14, 87.74, 76.12, 75.90, 75.84, 75.22, 74.49, 73.68, 73.58, 72.92, 72.20, 71.75, 70.63, 70.60, 70.36, 69.95, 69.57, 69.07, 65.29, 65.17, 64.92, 25.73, 21.41, 17.86, 17.14, 16.18, 16.15, 16.10, -4.84, -4.94; HRMS (ESI) Calcd for C 93 H 106 NO 20 SSi [M + NH4] + : 1616.6798, found: 1616.6796.

[0136] Compound 2-1

[0137]

[0138] Take a 100 mL two-necked reaction flask, add a magnetic stir bar, evacuate the flask together with the stir bar to vacuum, and then bake the reaction flask with a high-temperature blowtorch until no water mist appears on the inner wall of the flask. After the flask cools down to room temperature, add the pre-baked Molecular sieve (2.00 g), after being evacuated, the molecular sieve was baked and activated with a high-temperature spray gun. After the molecular sieve showed a phenomenon of being dispersed and adhering to the bottle wall, the baking was stopped. After waiting for the reaction flask to cool to room temperature again, the protective gas argon was introduced. Subsequently, anhydrous dichloromethane (20 mL) was added. Then, the glycosyl donor 1-6 (0.579 g, 1.00 mmol) and 2,4,6-tri-tert-butylpyrimidine (TTBP) (0.300 g, 1.20 mmol) were dissolved together in anhydrous dichloromethane (20 mL) and added to the reaction flask. After stirring at room temperature for 20 min, the reaction environment temperature was lowered to -78 °C. After 5 min, AgOTf (0.514 g, 2.00 mmol) pre-dissolved in anhydrous toluene (8 mL) was slowly added dropwise. Subsequently, a stoichiometric amount of p-TolSCl (133.3 μL, 1.00 mmol) was added, and the reaction was stirred for 5 min. Then, TLC monitoring was used, and it showed that the glycosyl donor 1-6 completely disappeared, indicating that the pre-activation reaction was complete. Immediately, the glycosyl acceptor 1-8 (0.373 g, 1.00 mmol) pre-dissolved in anhydrous dichloromethane (2 mL) was slowly added dropwise to the reaction solution. After the reaction environment temperature slowly rose to room temperature, TLC monitoring was used, and it showed that the reaction ended. Then, triethylamine (0.5 mL) was added dropwise to quench the reaction. The quenched reaction solution was filtered through diatomaceous earth to remove the molecular sieve, and the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (petroleum ether / ethyl acetate, volume ratio 5:1) to obtain the white solid 2-1 (0.761 g, 92%). R f = 0.30 (PE / EA volume ratio = 2:1); 1 H NMR (400 MHz, CDCl3) δ 8.06 - 8.02 (m, 2H), 7.95 - 7.91 (m, 2H), 7.58 - 7.51 (m, 2H), 7.43 - 7.35 (m, 8H), 7.34 - 7.30 (m, 1H), 7.19 - 7.14 (m, 5H), 5.61 - 5.57 (m, 1H), 5.18 (d, J = 3.4 Hz, 1H), 5.10 (dd, J = 3.7, 1.3 Hz, 1H), 5.02 (d, J = 10.7 Hz, 1H), 4.73 (d, J = 10.8 Hz, 1H), 4.48 (d, J = 12.2 Hz, 1H), 4.40 (d, J = 7.7 Hz, 1H), 4.33 - 4.24 (m, 2H), 4.18 (dd, J = 9.8, 3.4 Hz, 1H), 3.93 (dd, J = 9.9, 3.4 Hz, 1H), 3.81 - 3.74 (m, 2H), 3.72 (dd, J = 9.7, 3.4 Hz, 1H), 3.64 (s, 3H), 1.27 (d, J = 6.4 Hz, 3H), 0.91 (d, J = 6.5 Hz, 3H), 0.67 (s, 9H), -0.01 (s, 3H), -0.11 (s, 3H);13 C NMR (101 MHz, CDCl₃) δ 166.68, 138.52, 138.46, 133.13, 132.91, 130.46, 130.19, 129.89, 128.60, 128.40, 128.09, 128.02, 127.99, 127.96, 127.25, 105.17, 94.23, 78.23, 75.43, 75.19, 74.68, 74.45, 72.10, 69.65, 69.51, 65.32, 57.50, 25.77, 17.92, 16.70, 16.15, -4.73, -4.90; HRMS (ESI) Calcd for C 47 H 58 O 11 SiNa [M+Na] + : 849.3646, found: 849.3641.

[0139] Compound 3-1

[0140]

[0141] Compound 2-1 (0.828 g, 1.00 mmol) was added to a 50 mL plastic reaction tube and dissolved in anhydrous tetrahydrofuran (10 mL). Subsequently, the reaction tube was placed in an ice bath and stirred. After the reaction temperature dropped from room temperature to 0 °C, 70 wt% HF-Pyridine (2 mL) was slowly added dropwise. After waiting for about 5 min, the ice bath was removed, and the plastic reaction tube was placed in an oil bath pot set at 50 °C and stirred thoroughly for the reaction. After 4 h, TLC monitoring showed that the reaction was complete, and heating was stopped and the oil bath pot was removed. After the reaction temperature cooled to room temperature, ethyl acetate (10 mL) was added for dilution, and then the reaction tube was placed in an ice bath and stirred, and triethylamine (5 mL) was slowly added dropwise to quench. Subsequently, the reaction solution was concentrated to obtain a crude product. The crude product was further separated by column chromatography (petroleum ether / ethyl acetate, volume ratio 4:1) to obtain white solid 3-1 (0.642 g, 90%). R f = 0.40 (toluene / MeCN volume ratio = 7:1); 11H NMR (400 MHz, CDCl3) δ 8.12 - 8.07 (m, 2H), 7.94 - 7.90 (m, 2H), 7.59 - 7.52 (m, 2H), 7.44 - 7.33 (m, 9H), 7.21 - 7.11 (m, 5H), 5.65 (dd, J = 3.5, 1.0 Hz, 1H), 5.40 (d, J = 3.3 Hz, 1H), 5.07 (dd, J = 3.4, 1.3 Hz, 1H), 5.02 (d, J = 10.7 Hz, 1H), 4.65 (d, J = 10.7 Hz, 1H), 4.57 (d, J = 12.1 Hz, 1H), 4.43 (d, J = 7.7 Hz, 1H), 4.36 (d, J = 12.0 Hz, 1H), 4.33 - 4.27 (m, 1H), 4.11 (dd, J = 10.3, 3.4 Hz, 1H), 4.01 (dd, J = 9.9, 3.4 Hz, 1H), 3.86 - 3.77 (m, 2H), 3.77 (d, J = 3.3 Hz, 1H), 3.67 (s, 3H), 2.08 - 1.98 (m, 1H), 1.29 (d, J = 6.4 Hz, 3H), 0.92 (d, J = 6.5 Hz, 3H); 13 13C NMR (101 MHz, CDCl3) δ 166.62, 166.48, 138.43, 137.73, 133.37, 133.06, 130.21, 129.96, 129.87, 129.43, 128.54, 128.44, 128.39, 128.15, 128.12, 128.05, 127.78, 105.29, 92.28, 78.31, 73.56, 73.36, 71.92, 69.48, 68.73, 67.72, 65.06, 57.46, 16.60, 15.96; HRMS (ESI) Calcd for C 41 H 48 NO 11 [M + NH4] + : 730.3227, found: 730.3214.

[0142] Compound 2 - 7

[0143]

[0144] Take a 100 mL two - necked reaction flask, add a magnetic stir bar, evacuate the flask together with the stir bar to vacuum, then bake the reaction flask with a high - temperature blowtorch until no water mist appears on its inner wall. After the flask cools down to room temperature, add the pre - baked Molecular sieve (2.00 g), after being evacuated, the molecular sieve was baked and activated with a high-temperature spray gun. After the molecular sieve showed a phenomenon of dispersing and sticking to the bottle wall, the baking was stopped. After waiting for the reaction flask to cool to room temperature again, the protective gas argon was introduced. Subsequently, anhydrous dichloromethane (10 mL) was added. Then, the glycosyl donor 1-15 (160.0 mg, 100.0 μmol) and TTBP (60 mg, 240 μmol) were dissolved in anhydrous dichloromethane (10 μmol) and added to the reaction flask together.

[0145] After stirring at room temperature for 20 min, the reaction temperature was lowered to -78 °C. After 5 min, AgOTf (51.4 mg, 200.0 μmol) dissolved in anhydrous toluene (1 mL) was slowly added dropwise. Subsequently, a stoichiometric amount of p-TolSCl (13.3 μL, 100.0 μmol) was added, and the reaction was stirred for 5 min. Then, TLC monitoring was used, and it showed that the glycosyl donor 1-15 completely disappeared, indicating that the pre-activation reaction was complete. Immediately, the glycosyl acceptor 1-14 (72.4 mg, 90.0 μmol) dissolved in anhydrous dichloromethane (1 mL) was slowly added dropwise to the reaction solution. The temperature condition of -78 °C was maintained, and the reaction was carried out for 20 min. Then, the reaction temperature was slowly raised to room temperature and the reaction was maintained at room temperature for 20 min. TLC monitoring was used, and it showed that the glycosyl acceptor 1-14 completely disappeared, and a new spot, namely the hexasaccharide product, was formed.

[0146] Again, the reaction temperature was lowered to -78 °C. After 5 min, AgOTf (46.3 mg, 180.0 μmol) dissolved in anhydrous toluene (1 mL) was slowly added dropwise. Subsequently, a stoichiometric amount of p-TolSCl (12.0 μL, 90.0 μmol) was added, and the reaction was stirred for 5 min. Then, TLC monitoring was used, and it showed that the generated hexasaccharide product completely disappeared, indicating that the pre-activation reaction was complete. Immediately, the glycosyl acceptor 3-1 (64.2 mg, 90.0 μmol) dissolved in anhydrous dichloromethane (1 mL) was slowly added dropwise to the reaction solution. After the reaction temperature was slowly raised to room temperature, TLC monitoring was used, and it showed that the reaction ended. Then, triethylamine (0.5 mL) was added to quench the reaction. The quenched reaction solution was filtered through diatomaceous earth to remove the molecular sieve, and the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (toluene / acetonitrile, volume ratio 7:1) to obtain white solid 2-7 (229.5 mg, 80%). R f = 0.40 (Toluene / MeCN volume ratio = 7:1); 11H NMR (400 MHz, CDCl3) δ 8.10 (d, J = 7.2 Hz, 2H), 7.98 - 7.80 (m, 14H), 7.62 - 7.28 (m, 30H), 7.25 - 7.00 (m, 34H), 5.61 (d, J = 3.2 Hz, 1H), 5.26 (d, J = 2.6 Hz, 1H), 5.20 (dd, J = 7.4, 3.5 Hz, 2H), 5.15 (d, J = 2.4 Hz, 1H), 5.09 - 4.98 (m, 8H), 4.95 (dd, J = 8.5, 3.4 Hz, 2H), 4.92 (d, J = 2.9 Hz, 1H), 4.66 - 4.48 (m, 7H), 4.44 - 4.35 (m, 2H), 4.35 - 4.28 (m, 4H), 4.27 - 4.20 (m, 4H), 4.19 - 4.07 (m, 6H), 4.06 - 3.90 (m, 10H), 3.88 - 3.76 (m, 6H), 3.65 (s, 3H), 3.61 (dd, J = 9.7, 3.4 Hz, 1H), 1.29 (d, J = 6.4 Hz, 3H), 0.83 (d, J = 6.4 Hz, 6H), 0.73 (d, J = 6.5 Hz, 3H), 0.61 (q, J = 6.0 Hz, 21H), -0.16 (s, 3H), -0.25 (s, 3H); 1313C NMR (151 MHz, CDCl3) δ 166.58, 166.54, 166.47, 166.44, 166.43, 166.41, 138.47, 138.43, 138.25, 138.07, 138.04, 138.02, 137.97, 133.32, 133.17, 133.11, 133.08, 133.02, 132.83, 132.82, 130.21, 130.16, 130.08, 129.96, 129.93, 129.86, 129.84, 128.62, 128.49, 128.45, 128.33, 128.30, 128.28, 128.24, 128.21, 128.05, 127.92, 127.57, 127.53, 127.50, 127.45, 127.07, 105.15, 93.77, 93.05, 93.03, 92.95, 92.88, 92.86, 78.52, 75.84, 75.20, 74.45, 73.99, 73.85, 73.78, 73.54, 73.12, 72.81, 72.72, 72.69, 72.62, 72.09, 71.68, 70.58, 70.52, 70.45, 70.38, 70.30, 70.20, 70.17, 70.05, 69.58, 69.52, 69.10, 65.23, 64.98, 64.93, 64.86, 64.82, 57.43, 25.70, 17.83, 16.68, 16.13, 16.07, 16.05, 15.97, 15.95, -4.88, -4.98; HRMS (ESI) Calcd for C 167 H 186 N2O 41 Si[M + 2NH4] 2+ : 1452.1162, found: 1452.1168.

[0147] Compound 3 - 7

[0148]

[0149] Compound 2-7 (287.0 mg, 100.0 μmol) was added to a 50 mL plastic reaction tube and dissolved in anhydrous tetrahydrofuran (10 mL). Subsequently, the reaction tube was placed in an ice bath and stirred. After the reaction environment temperature dropped from room temperature to 0 °C, 70% HF-Pyridine (2 mL) was slowly added dropwise. After waiting for about 5 min, the ice bath was removed, and the plastic reaction tube was placed in an oil bath pot with a temperature set at 50 °C and stirred thoroughly for the reaction. After 4 h, TLC monitoring showed that the reaction was complete, and the heating was stopped and the oil bath pot was removed. After the reaction environment temperature cooled to room temperature, ethyl acetate (10 mL) was added for dilution. Subsequently, the reaction tube was placed in an ice bath and stirred, and triethylamine (5 mL) was slowly added dropwise to quench the reaction. Then the reaction solution was concentrated to obtain the crude product. The crude product was further separated by column chromatography (toluene / acetonitrile, volume ratio 7:1) to obtain white solid 3-7 (231.4 mg, 84%). R f = 0.30 (toluene / MeCN volume ratio = 7:1); 1 H NMR (400 MHz, CDCl3) δ 8.11 (d, J = 7.3 Hz, 2H), 7.99 - 7.85 (m, 14H), 7.63 - 7.48 (m, 10H), 7.46 - 7.30 (m, 22H), 7.25 - 7.01 (m, 30H), 6.97 (d, J = 7.3 Hz, 2H), 5.62 (d, J = 3.1 Hz, 1H), 5.26 (s, 1H), 5.21 (dd, J = 5.9, 3.5 Hz, 2H), 5.18 - 5.12 (m, 2H), 5.07 (s, 5H), 5.02 (t, J = 8.3 Hz, 4H), 4.82 (d, J = 2.9 Hz, 1H), 4.67 - 4.56 (m, 3H), 4.53 (d, J = 11.4 Hz, 4H), 4.49 - 4.40 (m, 2H), 4.37 - 4.30 (m, 3H), 4.24 (t, J = 12.9 Hz, 6H), 4.14 (d, J = 7.1 Hz, 3H), 4.10 (d, J = 3.5 Hz, 3H), 4.07 - 3.99 (m, 5H), 3.99 - 3.96 (m, 1H), 3.96 - 3.92 (m, 2H), 3.89 - 3.78 (m, 7H), 3.66 (s, 3H), 3.61 (dd, J = 10.1, 3.1 Hz, 1H), 1.84 (s, 1H), 1.30 (s, 3H), 0.84 (d, J = 6.4 Hz, 6H), 0.69 - 0.58 (m, 15H); 1313C NMR (101 MHz, CDCl3) δ 171.27, 166.59, 166.54, 166.53, 166.48, 166.43, 166.42, 138.45, 138.16, 138.09, 138.08, 138.07, 138.04, 137.99, 133.32, 133.17, 133.11, 133.03, 130.27, 130.26, 130.22, 130.17, 130.11, 130.10, 130.08, 129.98, 129.94, 129.91, 129.88, 129.84, 128.63, 128.50, 128.45, 128.40, 128.35, 128.31, 128.28, 128.24, 128.13, 128.05, 127.97, 127.85, 127.63, 127.57, 127.53, 105.17, 93.80, 93.04, 92.94, 92.87, 92.79, 91.64, 78.54, 75.85, 75.47, 74.25, 74.02, 73.82, 73.60, 73.53, 72.75, 72.71, 72.67, 72.64, 71.57, 70.60, 70.47, 70.40, 70.22, 70.19, 70.16, 70.08, 69.89, 69.74, 69.60, 69.12, 67.77, 65.25, 64.99, 64.89, 64.87, 64.71, 64.65, 57.44, 16.68, 16.14, 16.08, 15.98, 15.94, 15.92, 15.90; HRMS (ESI) Calcd for C 161 H 172 N2O41 [M + 2NH4] 2+ : 1395.0729, found: 1395.0753.

[0150] Compounds 4 - 7

[0151]

[0152] Compound 3-7 (96.4 mg, 35.0 μmol) was dissolved in a 50 mL volumetric flask containing a mixed solvent of MeOH / EtOAc (V / V = 1:1, 10 mL). Pd(OH)2 / C (96.4 mg) powder was added, and the mixture was stirred at room temperature for 6 h under a H2 atmosphere with a pressure of 0.4 MPa. TLC monitoring showed that the reaction was complete. Then the reaction solution was filtered through diatomaceous earth to remove the Pd(OH)2 / C powder, and the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (dichloromethane / methanol, volume ratio 20:1) to obtain white solid 4-7 (68.7 mg, 95%). R f = 0.40 (DCM / MeOH volume ratio = 10:1); 1 1H NMR (400 MHz, CDCl3) δ 8.23 - 8.17 (m, 2H), 8.15 - 8.02 (m, 14H), 7.67 - 7.39 (m, 24H), 5.52 (d, J = 3.1 Hz, 1H), 5.42 (d, J = 2.2 Hz, 1H), 5.40 - 5.37 (m, 2H), 5.36 (s, 2H), 5.32 (s, 2H), 5.21 (d, J = 3.9 Hz, 1H), 5.03 - 4.93 (m, 6H), 4.52 (q, J = 6.4 Hz, 1H), 4.46 (d, J = 7.1 Hz, 2H), 4.41 (s, 1H), 4.39 - 4.34 (m, 1H), 4.33 (d, J = 3.2 Hz, 1H), 4.31 (d, J = 2.3 Hz, 2H), 3.99 (td, J = 10.6, 3.7 Hz, 1H), 3.95 - 3.74 (m, 16H), 3.66 (s, 3H), 3.27 (s, 1H), 3.04 (d, J = 10.9 Hz, 1H), 2.81 (d, J = 9.8 Hz, 1H), 2.72 - 2.58 (m, 5H), 2.23 (s, 1H), 1.35 (s, 3H), 1.18 (d, J = 6.5 Hz, 3H), 1.14 (d, J = 6.5 Hz, 3H), 0.92 (d, J = 6.5 Hz, 3H), 0.88 (t, J = 5.8 Hz, 10H), 0.83 (d, J = 6.5 Hz, 3H); 1313C NMR (101 MHz, CDCl3) δ 167.85, 167.74, 167.67, 167.56, 167.52, 167.48, 166.63, 133.96, 133.90, 133.73, 133.69, 133.67, 133.64, 133.18, 132.47, 131.05, 130.33, 130.09, 130.06, 130.01, 129.88, 129.63, 129.62, 129.57, 129.37, 129.31, 128.98, 128.79, 128.76, 128.71, 128.50, 104.19, 100.81, 99.85, 98.94, 98.91, 98.79, 98.61, 82.45, 77.67, 77.04, 76.94, 73.70, 73.12, 72.93, 72.90, 72.78, 72.72, 71.93, 70.76, 69.92, 69.78, 69.47, 68.00, 67.95, 67.92, 66.55, 66.01, 65.89, 65.71, 65.67, 65.60, 65.52, 57.65, 16.41, 16.27, 16.01, 15.96, 15.91, 15.88; HRMS (ESI) Calcd for C 105 H 120 NO 41 [M+NH4] + : 2050.7336, found: 2050.7343.

[0153] Compound 6-7

[0154]

[0155] Compound 4-7 (51.7 mg, 25.0 μmol) was added to a 25 mL round-bottom flask containing anhydrous methanol (10 mL). After the compound was completely dissolved, a small amount of sodium methoxide powder was added until the pH of the reaction solution was approximately 10. Then, the round-bottom flask containing the reaction solution was placed in an oil bath at a temperature set to 50 °C, and the reaction was stirred thoroughly. After 2 h, TLC was used for monitoring, showing that the reaction was complete. The heating was stopped and the oil bath was removed. After the reaction solution was cooled to room temperature, cation exchange resin was added to the reaction solution until the pH of the reaction solution was neutral. Then, the adjusted reaction solution was filtered through diatomaceous earth to remove the resin, and the filtrate was concentrated to obtain the crude product. The crude product was further separated by column chromatography (dichloromethane / methanol, volume ratio 5:1) to obtain white solid 6-7 (31 mg, 91%). R f f = 0.30 (DCM / MeOH volume ratio = 3:1); 1HNMR(600MHz, MeOD) δ 5.04 - 4.96(m, 7H), 4.43 - 4.35(m, 7H), 4.18(d, J = 7.3Hz, 1H), 4.02 - 3.88(m, 18H), 3.86(dd, J = 10.1, 3.3Hz, 1H), 3.83(dd, J = 2.9, 1.1Hz, 1H), 3.80(dd, J = 10.2, 3.8Hz, 1H), 3.70(dd, J = 3.4, 1.2Hz, 1H), 3.67 - 3.58(m, 3H), 3.53(s, 3H), 1.32(d, J = 6.5Hz, 3H), 1.28 - 1.22(m, 21H); 13 C NMR(151MHz, MeOD) δ 105.75, 97.86, 97.73, 97.58, 80.11, 77.52, 77.44, 77.42, 77.39, 73.81, 71.63, 71.49, 70.46, 70.28, 70.21, 69.96, 69.28, 68.23, 68.18, 67.89, 67.41, 67.39, 67.33, 57.16, 16.77, 16.64, 16.62, 16.59; HRMS(ESI) Calcd for C 49 H 88 NO 33 [M + NH4] + : 1218.5239, found: 1218.5242.

[0156] Compound of formula 29

[0157] The washed reaction tube dedicated for the sulfation reaction was placed in an infrared drying oven and dried for 30 min. After removing the moisture remaining on the tube wall, the heating of the drying oven was turned off. After the temperature of the reaction tube dropped to room temperature, it was taken out. A magnetic stir bar was added to the reaction tube. After pumping to vacuum, argon, the protective gas, was introduced. Compound A’ (5 mg, 4.16 μmol) and sulfur trioxide-pyridine complex (168 mg, 1.06 mmol) were added to the reaction tube, and anhydrous DMF (2 mL) was quickly added, followed by stirring the reaction at room temperature. After 14 h, the reaction tube was placed in an ice bath, and triethylamine (0.2 mL) and methanol (0.2 mL) were successively added to quench the reaction. The quenched reaction solution was directly separated by a gel column (Sephadex LH-20, (V / V) H2O / MeOH = 1:1) to obtain the crude product of the sulfation intermediate. Then, this crude product was added to a 10 mL round-bottom flask containing 3 M NaOH (2 mL), and stirred at room temperature for 30 min. The reaction solution was separated and purified by a gel column (Sephadex LH-20, (V / V) H2O / MeOH = 1:1) to obtain the white solid of Formula 29 (11.0 mg, 91%). 1 HNMR(600MHz,D2O)δ5.45 - 5.42(m,6H),5.40(d,J=3.4Hz,1H),5.22(s,1H),4.97 - 4.93(m,8H),4.77 - 4.77(m,1H),4.67(dd,J=6.6,5.1Hz,1H),4.57 - 4.51(m,8H),4.46 - 4.35(m,10H),4.32(d,J=4.8Hz,1H),4.30 - 4.25(m,2H),4.20(t,J=5.0Hz,1H),3.45(s,3H),1.49(d,J=6.5Hz,3H),1.35(m,24H); 13 C NMR(151MHz,D2O)δ106.36,97.43,84.95,84.76,82.69,79.79,75.58,74.46,74.14,73.95,72.55,72.30,67.22,66.73,54.68,16.86,15.88,15.82,15.75;HRMS(ESIOrbitrap,negative)calcd for C 49 H 67 Na 14 O 84 S 17 3- [M - 3Na] 3- m / z 954.8269,found 954.8279.

[0158] Example 2

[0159] Compound 6-9 was prepared by a method similar to that of Example 1

[0160] Compound of formula 30

[0161] The washed reaction tube dedicated to the sulfation reaction was placed in an infrared drying oven and dried for 30 min. After removing the water remaining on the tube wall, the heating of the drying oven was turned off. After the temperature of the reaction tube dropped to room temperature, it was taken out. A magnetic stir bar was added to the reaction tube. After pumping to vacuum, the protective gas argon was introduced. Compound 6-9 (5 mg, 3.35 μmol) and sulfur trioxide-pyridine complex (166 mg, 1.05 mmol) were added to the reaction tube, and anhydrous DMF (2 mL) was quickly added, and the reaction was stirred at room temperature. After 14 h, the reaction tube was placed in an ice bath, and triethylamine (0.2 mL) and methanol (0.2 mL) were added in sequence to quench the reaction. The quenched reaction solution was directly separated by a gel column (Sephadex LH-20, (V / V) H2O / MeOH = 1:1) to obtain the crude sulfated intermediate product. Then the crude product was added to a 10 mL round bottom flask containing 3M NaOH (2 mL), and stirred at room temperature for 30 min. The reaction solution was separated and purified by a gel column (Sephadex LH-20, (V / V) H2O / MeOH = 1:1) to obtain the white solid of formula 30 (10.8 mg, 90%).

[0162] 1 H NMR (600 MHz, D2O) δ 5.48 - 5.39 (m, 9H), 5.23 (s, 1H), 4.98 - 4.92 (m, 10H), 4.78 - 4.77 (m, 1H), 4.68 (dd, J = 6.6, 5.1 Hz, 1H), 4.58 - 4.51 (m, 10H), 4.47 - 4.25 (m, 17H), 4.20 (t, J = 5.0 Hz, 1H), 3.46 (s, 3H), 1.50 (d, J = 6.5 Hz, 3H), 1.39 - 1.31 (m, 27H); 13 C NMR (151 MHz, D2O) δ 106.36, 97.42, 84.94, 84.78, 82.67, 80.38, 79.78, 75.56, 74.45, 74.22, 74.13, 73.94, 73.85, 73.80, 72.54, 72.30, 67.85, 67.49, 67.23, 66.73, 54.69, 16.87, 15.89, 15.83, 15.76, 15.71; HRMS (ESI Orbitrap, negative) calcd for C 61 H 83 Na 18O 104 S 21 3- [M-3Na] 3- m / z 1188.1172, found 1188.1188.

[0163] Example 3

[0164] Compound Formula 37

[0165] Put the washed reaction tube dedicated to the sulfation reaction into an infrared drying oven and dry it for 30 min. After removing the water remaining on the tube wall, turn off the heating of the drying oven. After the temperature of the reaction tube drops to room temperature, take it out. Add a magnetic stir bar into the reaction tube. After pumping to vacuum, introduce the protective gas argon. Add compound 13-7 (15 mg, 9.27 μmol) and sulfur trioxide-pyridine complex (287 mg, 1.81 mmol) into the reaction tube, quickly add anhydrous DMF (2 mL), and stir the reaction at room temperature. After 7 h, place the reaction tube in an ice bath, and sequentially add triethylamine (0.2 mL) and methanol (0.2 mL) to quench the reaction. The quenched reaction solution was directly separated by a gel column (Sephadex LH-20, (V / V) DCM / MeOH = 1:1) to obtain a crude product of the sulfation intermediate. Then add this crude product into a 10 mL round-bottom flask containing 3M NaOH (2 mL) and stir at room temperature for 4 h. The reaction solution was separated and purified by a gel column (Sephadex LH-20, (V / V) H2O / MeOH = 1:1) to obtain white solid 37 (22.2 mg, 95%). 1 HNMR(600MHz, D2O) δ 5.45(dd, J = 9.3, 3.8Hz, 3H), 5.38(d, J = 3.7Hz, 1H), 5.32(dd, J = 9.7, 3.8Hz, 3H), 4.86 - 4.80(m, 4H), 4.72 - 4.66(m, 3H), 4.66 - 4.56(m, 8H), 4.51(dq, J = 20.4, 6.7Hz, 2H), 4.47 - 4.37(m, 4H), 4.33 - 4.21(m, 8H), 4.15(d, J = 3.1Hz, 3H), 3.93(q, J = 6.5Hz, 1H), 3.57(s, 3H), 1.48 - 1.40(m, 12H), 1.34 - 1.29(m, 12H); 1313C NMR (151 MHz, D2O) δ 102.11, 98.68, 98.60, 98.58, 98.50, 94.97, 94.69, 94.47, 79.48, 79.30, 78.92, 78.51, 77.69, 75.86, 74.85, 73.88, 73.78, 73.69, 73.42, 73.26, 73.24, 73.11, 72.64, 72.42, 72.40, 72.37, 70.92, 70.78, 69.51, 69.36, 69.23, 67.84, 67.82, 67.75, 67.09, 67.06, 66.79, 57.08, 15.83, 15.59, 15.54, 15.52, 15.36, 15.35, 15.32, 15.19; HRMS (ESI Orbitrap, negative) calcd for C 49 H 71 Na 11 O 72 S 13 2- [M - 2Na] 2- m / z 1239.8575, found 1239.8586.

[0166] Example 4

[0167] Compound of formula 58

[0168] The washed reaction tube dedicated for the sulfation reaction was placed in an infrared drying oven and dried for 30 min. After removing the moisture remaining on the tube wall, the heating of the drying oven was turned off. After the temperature of the reaction tube dropped to room temperature, it was taken out. A magnetic stir bar was added to the reaction tube. After pumping to vacuum, argon gas as the protective gas was introduced. Compound 15 - 6 (5 mg, 4.74 μmol) and sulfur trioxide - pyridine complex (170 mg, 1.07 mmol) were added to the reaction tube, and anhydrous DMF (2 mL) was quickly added, and the reaction was stirred at room temperature. After 14 h, the reaction tube was placed in an ice bath, and triethylamine (0.2 mL) and methanol (0.2 mL) were added successively to quench the reaction. The quenched reaction solution was directly separated by a gel column (Sephadex LH - 20, V / V) H2O / MeOH = 1:1) to obtain a crude product of the sulfation intermediate. Then, this crude product was added to a 10 mL round - bottom flask containing 3M NaOH (2 mL), and stirred at room temperature for 30 min. The reaction solution was separated and purified by a gel column (Sephadex LH - 20, V / V) H2O / MeOH = 1:1) to obtain a white solid of formula 58 (11.6 mg, 95%). 11H NMR (600 MHz, D2O) δ 5.50 - 5.43 (m, 4H), 5.39 (dd, J = 7.1, 3.6 Hz, 2H), 5.00 - 4.92 (m, 7H), 4.69 (dd, J = 10.8, 3.4 Hz, 2H), 4.66 - 4.52 (m, 10H), 4.47 - 4.37 (m, 6H), 4.33 - 4.30 (m, 3H), 3.94 (q, J = 6.5 Hz, 1H), 3.57 (s, 3H), 1.46 (d, J = 6.4 Hz, 3H), 1.43 (dd, J = 6.6, 4.9 Hz, 6H), 1.40 - 1.36 (m, 9H), 1.34 (d, J = 6.6 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 102.15, 98.65, 98.62, 98.53, 96.34, 96.32, 96.23, 79.82, 79.71, 79.65, 79.62, 79.57, 78.78, 77.77, 75.87, 74.00, 73.87, 73.80, 73.65, 73.58, 72.56, 72.46, 72.44, 72.40, 72.34, 72.28, 72.17, 72.06, 70.92, 68.06, 68.00, 67.50, 67.43, 67.39, 66.69, 57.06, 16.04, 16.02, 15.98, 15.87, 15.84, 15.72; HRMS (ESI Orbitrap, negative) calcd for C 43 H 59 Na 12 O 74 S 15 3- [M - 3Na] 3- m / z 838.1818, found 838.1831.

[0169] Examples 5 - 61

[0170] Compound 1

[0171] 11H NMR (400 MHz, D2O) δ 5.35 (d, J = 3.9 Hz, 1H), 4.75 (dd, J = 10.5, 3.2 Hz, 1H), 4.61 - 4.51 (m, 2H), 4.45 (d, J = 7.9 Hz, 1H), 4.32 (dd, J = 9.9, 7.9 Hz, 1H), 4.18 (d, J = 2.2 Hz, 1H), 4.01 (d, J = 3.1 Hz, 1H), 3.85 (dd, J = 9.9, 3.1 Hz, 1H), 3.78 (q, J = 6.4 Hz, 1H), 3.52 (s, 3H), 1.27 (d, J = 6.5 Hz, 3H), 1.20 (d, J = 6.6 Hz, 3H); 13 13C NMR (101 MHz, D2O) δ 102.46, 93.68, 76.23, 76.15, 74.90, 72.39, 70.55, 67.65, 66.57, 57.38, 15.37, 15.20; HRMS (ESI Orbitrap, negative) calcd for C 13 H 21 Na2O 18 S3 - [M - Na] - m / z 606.9691, found 606.9691.

[0172] Compound 2

[0173] 1 1H NMR (400 MHz, D2O) δ 5.41 - 5.39 (m, 2H), 4.75 (dd, J = 10.5, 3.2 Hz, 1H), 4.64 - 4.49 (m, 5H), 4.38 (dd, J = 9.9, 7.9 Hz, 1H), 4.27 - 4.18 (m, 2H), 4.12 (d, J = 2.6 Hz, 1H), 4.07 (d, J = 3.1 Hz, 1H), 3.90 (dd, J = 9.9, 3.1 Hz, 1H), 3.84 (q, J = 6.5 Hz, 1H), 3.58 (s, 3H), 1.33 (d, J = 6.4 Hz, 3H), 1.34 - 1.26 (m, 6H); 13 13C NMR (101 MHz, D2O) δ 102.47, 95.09, 93.50, 76.27, 75.95, 75.05, 73.95, 73.06, 72.34, 70.62, 70.55, 69.31, 67.59, 66.68, 66.47, 57.34, 15.39, 15.27; HRMS (ESI Orbitrap, negative) calcd for C 19 H30 Na3O 25 S4 - [M-Na] - m / z 854.9658, found 854.9663.

[0174] Compound 3

[0175] 1 1H NMR (600 MHz, D2O) δ 5.43 - 5.36 (m, 3H), 4.74 (dd, J = 10.5, 3.2 Hz, 1H), 4.63 - 4.49 (m, 6H), 4.46 (q, J = 6.6 Hz, 1H), 4.38 (dd, J = 9.9, 7.9 Hz, 1H), 4.24 (d, J = 3.3 Hz, 1H), 4.19 (dd, J = 10.4, 3.1 Hz, 1H), 4.16 - 4.10 (m, 3H), 4.07 (d, J = 3.1 Hz, 1H), 3.89 (dd, J = 9.9, 3.1 Hz, 1H), 3.83 (q, J = 6.5 Hz, 1H), 3.57 (s, 3H), 1.33 (d, J = 6.4 Hz, 3H), 1.31 - 1.25 (m, 9H); 13 13C NMR (101 MHz, D2O) δ 102.43, 95.32, 94.86, 93.59, 76.22, 76.07, 74.98, 74.34, 73.60, 73.07, 72.99, 72.27, 70.57, 70.50, 69.43, 69.18, 67.62, 66.66, 66.57, 66.45, 57.26, 15.34, 15.30, 15.25; HRMS (ESI Orbitrap, negative) calcd for C 25 H 39 Na3O 32 S5 2- [M - 2Na] 2- m / z 539.9866, found 539.9866.

[0176] Compound 4

[0177] 11H NMR (400 MHz, D2O) δ 5.43 - 5.33 (m, 4H), 4.72 (dd, J = 10.5, 3.2 Hz, 1H), 4.62 - 4.41 (m, 9H), 4.36 (dd, J = 9.9, 7.9 Hz, 1H), 4.24 - 4.09 (m, 7H), 4.05 (d, J = 3.1 Hz, 1H), 3.88 (dd, J = 9.9, 3.1 Hz, 1H), 3.81 (q, J = 6.5 Hz, 1H), 3.56 (s, 3H), 1.33 - 1.23 (m, 15H); 13 13C NMR (101 MHz, D2O) δ 102.43, 95.33, 95.11, 94.90, 93.62, 76.21, 76.11, 74.98, 74.34, 74.00, 73.66, 73.06, 73.03, 72.96, 72.27, 70.57, 70.48, 69.44, 69.33, 69.20, 67.62, 66.65, 66.58, 66.56, 66.44, 57.24, 15.31, 15.24; HRMS (ESI Orbitrap, negative) calcd for C 31 H 48 Na4O 39 S6 2- [M - 2Na] 2- m / z 663.9849, found 663.9853.

[0178] Compound 5

[0179] 1 1H NMR (400 MHz, D2O) δ 5.39 - 5.36 (m, 5H), 4.73 (dd, J = 10.5, 3.2 Hz, 1H), 4.64 - 4.41 (m, 11H), 4.37 (dd, J = 9.8, 7.9 Hz, 1H), 4.25 - 4.09 (m, 9H), 4.06 (d, J = 3.1 Hz, 1H), 3.88 (dd, J = 9.9, 3.1 Hz, 1H), 3.82 (q, J = 6.5 Hz, 1H), 3.56 (s, 3H), 1.35 - 1.23 (m, 18H); 13¹³C NMR (101 MHz, D₂O) δ 102.46, 95.32, 95.13, 95.11, 94.91, 93.62, 76.25, 76.11, 75.02, 74.32, 74.05, 74.01, 73.68, 73.10, 73.06, 73.05, 73.00, 72.31, 70.60, 70.52, 69.44, 69.35, 69.22, 67.65, 66.68, 66.61, 66.46, 57.28, 15.36, 15.28; HRMS (ESI Orbitrap, negative) calcd for C 37 H 57 Na₅O 46 S₇ 2- [M - 2Na] 2- m / z 787.9833, found 787.9840.

[0180] Compound 6

[0181] 1 ¹H NMR (400 MHz, D₂O) δ 5.38 - 5.32 (m, 6H), 4.70 (dd, J = 10.5, 3.2 Hz, 1H), 4.58 - 4.49 (m, 8H), 4.47 - 4.39 (m, 5H), 4.34 (dd, J = 9.8, 7.9 Hz, 1H), 4.20 (d, J = 3.3 Hz, 1H), 4.17 - 4.06 (m, 10H), 4.03 (d, J = 3.1 Hz, 1H), 3.86 (dd, J = 9.9, 3.1 Hz, 1H), 3.79 (q, J = 6.6 Hz, 1H), 3.53 (s, 3H), 1.30 - 1.22 (m, 21H); 13 ¹³C NMR (101 MHz, D₂O) δ 102.43, 95.32, 95.16, 95.11, 94.92, 93.62, 76.22, 76.10, 74.98, 74.33, 74.07, 74.00, 73.68, 73.06, 73.03, 73.01, 72.96, 72.27, 70.57, 70.49, 69.44, 69.35, 69.21, 67.63, 66.66, 66.59, 66.44, 57.24, 15.33, 15.25, 15.24; HRMS (ESI Orbitrap, negative) calcd for C 43 H 66 Na₆O 53 S₈ 2- [M - 2Na] 2-m / z 911.9816, found 911.9827.

[0182] Compound 7

[0183] 1 H NMR (600 MHz, D2O) δ 5.39 - 5.37 (m, 7H), 4.73 (dd, J=10.5, 3.2 Hz, 1H), 4.62 - 4.52 (m, 9H), 4.51 - 4.43 (m, 6H), 4.37 (dd, J=9.9, 7.9 Hz, 1H), 4.23 (d, J=4.0 Hz, 1H), 4.18 (dd, J=10.3, 3.1 Hz, 1H), 4.17 - 4.08 (m, 11H), 4.06 (d, J=3.1 Hz, 1H), 3.89 (dd, J=9.9, 3.1 Hz, 1H), 3.82 (q, J=6.8 Hz, 1H), 3.56 (s, 3H), 1.33 - 1.25 (m, 24H); 13 C NMR (151 MHz, D2O) δ 102.45, 95.33, 95.18, 95.12, 94.93, 93.63, 76.23, 76.11, 75.00, 74.33, 74.08, 74.00, 73.69, 73.08, 73.05, 73.02, 72.98, 72.29, 70.58, 70.50, 69.45, 69.37, 69.35, 69.22, 67.64, 66.67, 66.60, 66.57, 66.46, 57.26, 15.35, 15.33, 15.27, 15.26; HRMS (ESI Orbitrap, negative) calcd for C 49 H 75 Na7O 60 S9 2- [M - 2Na] 2- m / z 1035.9799, found 1035.9816.

[0184] Compound 8

[0185] 11H NMR (600 MHz, D2O) δ 5.39 - 5.36 (m, 8H), 4.72 (dd, J = 10.5, 3.2 Hz, 1H), 4.60 - 4.51 (m, 10H), 4.50 - 4.41 (m, 7H), 4.37 (dd, J = 9.9, 7.9 Hz, 1H), 4.24 - 4.21 (m, 1H), 4.18 (dd, J = 10.3, 3.1 Hz, 1H), 4.16 - 4.07 (m, 13H), 4.07 - 4.04 (m, 1H), 3.88 (dd, J = 9.9, 3.0 Hz, 1H), 3.84 - 3.79 (m, 1H), 3.56 (s, 3H), 1.33 - 1.24 (m, 27H); 13 13C NMR (151 MHz, D2O) δ 102.44, 95.33, 95.18, 95.11, 94.94, 93.66, 76.22, 76.15, 74.99, 74.35, 74.09, 74.08, 74.00, 73.70, 73.08, 73.04, 73.02, 72.97, 72.28, 70.58, 70.50, 69.45, 69.37, 69.22, 67.65, 66.66, 66.59, 66.46, 57.24, 15.35, 15.33, 15.32, 15.27, 15.25; HRMS (ESI Orbitrap, negative) calcd for C 55 H 84 Na8O 67 S 10 2- [M - 2Na] 2- m / z 1159.9783, found 1159.9796.

[0186] Compound 9

[0187] 1 1H NMR (600 MHz, D2O) δ 5.39 - 5.36 (m, 9H), 4.73 (dd, J = 10.5, 3.2 Hz, 1H), 4.60 - 4.51 (m, 11H), 4.50 - 4.41 (m, 8H), 4.37 (dd, J = 9.9, 7.9 Hz, 1H), 4.24 - 4.21 (m, 1H), 4.19 - 4.09 (m, 16H), 4.06 (d, J = 3.1 Hz, 1H), 3.88 (dd, J = 9.9, 3.1 Hz, 1H), 3.84 - 3.79 (m, 1H), 3.56 (s, 3H), 1.33 - 1.25 (m, 30H); 13¹³C NMR (151 MHz, D₂O) δ 102.44, 95.34, 95.18, 95.12, 94.94, 93.66, 76.22, 76.15, 75.00, 74.35, 74.09, 74.01, 73.70, 73.08, 73.04, 73.02, 72.97, 72.28, 70.58, 70.50, 69.45, 69.37, 69.34, 69.22, 67.65, 66.66, 66.57, 66.46, 57.24, 15.35, 15.32, 15.27, 15.25; HRMS (ESI Orbitrap, negative) calcd for C 61 H 93 Na₉O 74 S 11 2- [M - 2Na] 2- m / z 1283.9766, found 1283.9782.

[0188] Compound 10

[0189] 1 ¹H NMR (600 MHz, D₂O) δ 5.39 - 5.36 (m, 10H), 4.72 (dd, J = 10.5, 3.2 Hz, 1H), 4.61 - 4.43 (m, 21H), 4.37 (dd, J = 9.9, 7.9 Hz, 1H), 4.23 (d, J = 3.4 Hz, 1H), 4.22 - 4.07 (m, 18H), 4.05 (d, J = 3.2 Hz, 1H), 3.88 (dd, J = 9.9, 3.1 Hz, 1H), 3.82 (q, J = 6.6 Hz, 1H), 3.56 (s, 3H), 1.32 - 1.25 (m, 33H); 13 ¹³C NMR (151 MHz, D₂O) δ 102.44, 95.34, 95.18, 95.11, 94.94, 93.67, 76.22, 76.16, 74.99, 74.35, 74.09, 74.00, 73.70, 73.08, 73.04, 73.02, 72.97, 72.28, 70.58, 70.50, 69.45, 69.37, 69.34, 69.22, 67.66, 66.66, 66.59, 66.57, 66.46, 57.24, 15.35, 15.32, 15.27, 15.25; HRMS (ESI Orbitrap, negative) calcd for C 67 H 102 Na₉O 81 S12 3- [M - 3Na] 3- m / z 930.9869, found 930.9884.

[0190] Compound 11

[0191] 1 H NMR(600MHz, D2O) δ 5.38 - 5.36(m, 11H), 4.72(dd, J=10.5, 3.2Hz, 1H), 4.61 - 4.42(m, 23H), 4.36(dd, J=9.8, 8.0Hz, 1H), 4.24 - 4.04(m, 22H), 3.88(dd, J=10.0, 3.0Hz, 1H), 3.82(q, J=6.5Hz, 1H), 3.57 - 3.54(m, 3H), 1.32 - 1.25(m, 36H); 13 C NMR(151MHz, D2O) δ 103.21, 96.11, 95.96, 95.88, 95.71, 94.46, 76.99, 76.95, 75.77, 75.13, 74.86, 74.77, 74.47, 73.85, 73.79, 73.75, 73.05, 71.35, 71.27, 70.23, 70.14, 69.99, 68.43, 67.43, 67.36, 67.23, 58.01, 16.12, 16.09, 16.04, 16.02; HRMS(ESI Orbitrap, negative) calcd for C 73 H 111 Na 10 O 88 S 13 3- [M - 3Na] 3- m / z 1013.6524, found 1013.6536.

[0192] Compound 12

[0193] 11H NMR (400 MHz, D2O) δ 5.24 (d, J = 4.0 Hz, 1H), 4.92 (d, J = 3.0 Hz, 1H), 4.76 (d, J = 3.0 Hz, 1H), 4.66 (dd, J = 10.6, 3.0 Hz, 1H), 4.52 (q, J = 6.6 Hz, 1H), 4.41 (d, J = 7.9 Hz, 1H), 4.00 - 3.90 (m, 2H), 3.86 (dd, J = 10.0, 3.0 Hz, 1H), 3.68 (dd, J = 10.1, 7.9 Hz, 1H), 3.60 (s, 3H), 1.36 (d, J = 6.4 Hz, 3H), 1.29 (d, J = 6.5 Hz, 3H); 13 13C NMR (101 MHz, D2O) δ 103.60, 97.11, 79.12, 77.74, 77.61, 75.42, 69.99, 68.87, 66.55, 66.39, 57.25, 15.92, 15.77; HRMS (ESI Orbitrap, negative) calcd for C 13 H 21 NaO 18 S3 2- [M - 2Na] 2- m / z 291.9899, found 291.9899.

[0194] Compound 13

[0195] 1 1H NMR (400 MHz, D2O) δ 5.09 - 5.06 (m, 2H), 4.81 (d, J = 3.0 Hz, 1H), 4.71 (s, 1H), 4.64 (d, J = 2.9 Hz, 1H), 4.54 (dd, J = 10.5, 3.0 Hz, 1H), 4.45 (q, J = 6.5 Hz, 1H), 4.38 (q, J = 6.5 Hz, 1H), 4.29 (d, J = 7.9 Hz, 1H), 3.98 (dd, J = 10.4, 2.9 Hz, 1H), 3.87 - 3.79 (m, 3H), 3.72 (dd, J = 10.0, 3.0 Hz, 1H), 3.55 (dd, J = 10.0, 7.8 Hz, 1H), 3.49 (s, 3H), 1.25 (d, J = 6.4 Hz, 3H), 1.21 (d, J = 6.5 Hz, 3H), 1.16 (d, J = 6.5 Hz, 3H); 1313C NMR (101 MHz, D2O) δ 103.71, 98.55, 97.63, 79.34, 79.14, 79.08, 78.30, 75.59, 75.46, 69.96, 69.00, 67.08, 66.68, 66.51, 57.25, 15.83, 15.68, 15.65; HRMS (ESI Orbitrap, negative) calcd for C 19 H 30 Na2O 25 S4 2- [M - 2Na] 2- m / z 415.9883, found 415.9882.

[0196] Compound 14

[0197] 1 1H NMR (400 MHz, D2O) δ 5.22 - 5.13 (m, 3H), 4.92 (d, J = 3.0 Hz, 1H), 4.82 - 4.80 (m, 2H), 4.74 (d, J = 2.9 Hz, 1H), 4.65 (dd, J = 10.6, 3.1 Hz, 1H), 4.57 - 4.46 (m, 3H), 4.40 (d, J = 7.9 Hz, 1H), 4.09 (td, J = 10.6, 2.9 Hz, 2H), 3.97 - 3.89 (m, 4H), 3.83 (dd, J = 10.1, 3.0 Hz, 1H), 3.68 - 3.63 (m, 1H), 3.60 (s, 3H), 1.36 - 1.26 (m, 12H); 13 13C NMR (101 MHz, D2O) δ 103.72, 98.59, 98.50, 97.65, 79.52, 79.42, 79.38, 79.03, 78.34, 76.30, 75.67, 75.50, 69.97, 69.01, 67.16, 66.65, 66.61, 66.53, 57.27, 15.84, 15.66, 15.58; HRMS (ESI Orbitrap, negative) calcd for C 25 H 39 Na3O 32 S5 2- [M - 2Na] 2- m / z 539.9866, found 539.9863.

[0198] Compound 15

[0199] 11H NMR (600 MHz, D2O) δ 5.10 - 5.03 (m, 4H), 4.82 (d, J = 3.0 Hz, 1H), 4.71 - 4.70 (m, 3H), 4.65 (d, J = 3.0 Hz, 1H), 4.55 (dd, J = 10.6, 3.1 Hz, 1H), 4.48 - 4.37 (m, 4H), 4.30 (d, J = 7.9 Hz, 1H), 4.02 - 3.94 (m, 3H), 3.83 (m, 5H), 3.73 (dd, J = 10.1, 3.0 Hz, 1H), 3.56 (dd, J = 10.1, 7.9 Hz, 1H), 3.50 (s, 3H), 1.25 (d, J = 6.4 Hz, 3H), 1.22 - 1.20 (m, 9H), 1.17 (d, J = 6.5 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 103.70, 98.58, 98.52, 98.47, 97.68, 79.76, 79.54, 79.52, 79.47, 79.42, 79.37, 79.01, 78.32, 76.33, 76.32, 76.26, 75.73, 75.70, 75.48, 69.95, 68.98, 67.21, 67.15, 66.72, 66.64, 66.59, 66.53, 57.25, 15.82, 15.63, 15.55, 15.52; HRMS (ESI Orbitrap, negative) calcd for C 31 H 48 Na4O 39 S6 2- [M - 2Na] 2- m / z 663.9849, found 663.9845.

[0200] Compound 16

[0201] 11H NMR (600 MHz, D2O) δ 5.18 - 5.11 (m, 5H), 4.90 (d, J = 3.1 Hz, 1H), 4.78 - 4.77 (m, 4H), 4.72 (d, J = 3.0 Hz, 1H), 4.63 (dd, J = 10.5, 3.0 Hz, 1H), 4.57 - 4.43 (m, 5H), 4.38 (d, J = 7.9 Hz, 1H), 4.10 - 4.02 (m, 4H), 3.94 - 3.87 (m, 6H), 3.81 (dd, J = 10.1, 3.0 Hz, 1H), 3.63 (dd, J = 10.1, 7.9 Hz, 1H), 3.57 (s, 3H), 1.33 (d, J = 6.4 Hz, 3H), 1.31 - 1.27 (m, 12H), 1.25 (d, J = 6.5 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 103.70, 98.58, 98.56, 98.53, 98.49, 97.68, 79.80, 79.78, 79.54, 79.49, 79.38, 79.01, 78.32, 76.37, 76.28, 76.26, 75.75, 75.48, 69.95, 68.98, 67.22, 67.16, 66.72, 66.68, 66.64, 66.59, 66.53, 57.25, 15.82, 15.64, 15.55, 15.51; HRMS (ESI Orbitrap, negative) calcd for C 37 H 57 Na5O 46 S7 2- [M - 2Na] 2- m / z 787.9833, found 787.9840.

[0202] Compound 17

[0203] 11H NMR (600 MHz, D2O) δ 5.21 - 5.08 (m, 6H), 4.90 (d, J = 3.1 Hz, 1H), 4.78 - 4.77 (m, 5H), 4.73 (d, J = 3.0 Hz, 1H), 4.63 (dd, J = 10.5, 3.1 Hz, 1H), 4.56 - 4.45 (m, 6H), 4.38 (d, J = 7.9 Hz, 1H), 4.09 - 4.04 (m, 5H), 3.92 - 3.88 (m, 7H), 3.81 (dd, J = 10.1, 3.0 Hz, 1H), 3.63 (dd, J = 10.1, 7.9 Hz, 1H), 3.58 (s, 3H), 1.34 - 1.32 (m, 3H), 1.31 - 1.27 (m, 15H), 1.25 (d, J = 6.5 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 103.70, 98.59, 98.55, 98.53, 98.52, 98.47, 97.66, 79.82, 79.78, 79.54, 79.48, 79.36, 79.02, 78.32, 76.36, 76.32, 76.27, 76.25, 75.72, 75.48, 69.95, 68.98, 67.22, 67.15, 66.72, 66.68, 66.63, 66.59, 66.52, 57.25, 15.82, 15.63, 15.55, 15.51; HRMS (ESI Orbitrap, negative) calcd for C 43 H 66 Na6O 53 S8 2- [M - 2Na] 2- m / z 911.9816, found 911.9818.

[0204] Compound 18

[0205] 1 1H NMR (400 MHz, D2O) δ 5.19 - 5.10 (m, 7H), 4.91 (d, J = 3.1 Hz, 1H), 4.80 - 4.79 (m, 6H), 4.73 (d, J = 3.1 Hz, 1H), 4.64 (dd, J = 10.5, 3.1 Hz, 1H), 4.56 - 4.45 (m, 7H), 4.39 (d, J = 7.8 Hz, 1H), 4.12 - 4.03 (m, 6H), 3.93 - 3.88 (m, 8H), 3.82 (dd, J = 10.1, 3.0 Hz, 1H), 3.64 (dd, J = 10.0, 7.9 Hz, 1H), 3.58 (s, 3H), 1.35 - 1.25 (m, 24H);13 C NMR (101 MHz, D2O) δ 103.71, 98.58, 98.52, 98.47, 97.65, 79.81, 79.77, 79.54, 79.47, 79.37, 79.02, 78.32, 76.37, 76.31, 76.27, 76.25, 75.71, 75.50, 69.96, 69.00, 67.23, 67.16, 66.73, 66.69, 66.64, 66.60, 66.53, 57.26, 15.83, 15.65, 15.57, 15.53; HRMS (ESI Orbitrap, negative) calcd for C 49 H 75 Na7O 60 S9 2- [M - 2Na] 2- m / z 1035.9799, found 1035.9805.

[0206] Compound 19

[0207] 1 H NMR (600 MHz, D2O) δ 5.19 - 5.10 (m, 8H), 4.90 (d, J = 3.2 Hz, 1H), 4.78 - 4.77 (m, 7H), 4.72 (d, J = 3.1 Hz, 1H), 4.63 (dd, J = 10.5, 3.0 Hz, 1H), 4.60 - 4.36 (m, 9H), 4.15 - 4.02 (m, 7H), 3.99 - 3.87 (m, 9H), 3.81 (dd, J = 10.1, 3.0 Hz, 1H), 3.63 (dd, J = 10.0, 7.9 Hz, 1H), 3.57 (s, 3H), 1.33 (d, J = 6.4 Hz, 3H), 1.31 - 1.24 (m, 24H); 13 C NMR (151 MHz, D2O) δ 104.47, 99.33, 99.25, 98.45, 80.59, 80.54, 80.31, 80.26, 80.15, 79.78, 79.08, 77.14, 77.10, 77.04, 76.52, 76.25, 70.72, 69.74, 68.00, 67.93, 67.49, 67.45, 67.41, 67.36, 67.30, 58.01, 16.59, 16.40, 16.31, 16.28; HRMS (ESI Orbitrap, negative) calcd for C 55 H 84 Na7O 67 S 103- [M - 3Na] 3- m / z 765.6558, found 765.6561.

[0208] Compound 20

[0209] 1 H NMR(600 MHz, D2O) δ 5.21 - 5.04(m, 9H), 4.89(d, J = 3.3 Hz, 1H), 4.78 - 4.77(m, 8H), 4.72(d, J = 3.1 Hz, 1H), 4.62(dd, J = 10.5, 3.0 Hz, 1H), 4.56 - 4.45(m, 9H), 4.37(d, J = 7.9 Hz, 1H), 4.09 - 4.01(m, 8H), 3.91 - 3.87(m, 10H), 3.80(dd, J = 10.1, 3.0 Hz, 1H), 3.63(dd, J = 10.0, 7.9 Hz, 1H), 3.57(s, 3H), 1.33(d, J = 6.4 Hz, 3H), 1.33 - 1.24(m, 24H), 1.24(d, J = 6.5 Hz, 3H); 13 C NMR(151 MHz, D2O) δ 103.69, 98.57, 98.55, 98.52, 98.49, 97.72, 79.85, 79.82, 79.76, 79.52, 79.39, 78.97, 78.27, 76.38, 76.34, 76.27, 75.79, 75.48, 69.95, 68.95, 67.22, 67.16, 67.14, 66.72, 66.67, 66.64, 66.58, 66.53, 66.18, 57.22, 15.81, 15.62, 15.53, 15.50; HRMS(ESI Orbitrap, negative) calcd for C 61 H 93 Na8O 74 S 11 3- [M - 3Na] 3- m / z 848.3213, found 848.3222.

[0210] Compound 21

[0211] 11H NMR (400 MHz, D2O) δ 5.19 - 5.07 (m, 10H), 4.88 (d, J = 3.2 Hz, 1H), 4.81 - 4.80 (m, 2H), 4.77 - 4.76 (m, 7H), 4.71 (d, J = 3.0 Hz, 1H), 4.61 (dd, J = 10.6, 3.0 Hz, 1H), 4.53 - 4.42 (m, 10H), 4.36 (d, J = 7.9 Hz, 1H), 4.09 - 3.97 (m, 9H), 3.91 - 3.85 (m, 11H), 3.79 (dd, J = 10.0, 2.9 Hz, 1H), 3.62 (dd, J = 8.3, 1.7 Hz, 1H), 3.56 (s, 3H), 1.33 - 1.22 (m, 33H); 13 13C NMR (101 MHz, D2O) δ 103.71, 98.59, 98.54, 98.48, 97.67, 79.83, 79.77, 79.76, 79.53, 79.49, 79.38, 79.35, 79.00, 78.30, 76.37, 76.32, 76.26, 75.73, 75.49, 69.95, 68.97, 67.22, 67.16, 66.68, 66.64, 66.59, 66.52, 57.23, 15.82, 15.64, 15.54, 15.51; HRMS (ESI Orbitrap, negative) calcd for C 67 H 102 Na9O 81 S 12 3- [M - 3Na] 3- m / z 930.9869, found 930.9879.

[0212] Compound 22

[0213] 1 1H NMR (400 MHz, D2O) δ 5.17 - 5.06 (m, 11H), 4.88 (d, J = 3.2 Hz, 1H), 4.77 - 4.76 (m, 10H), 4.71 (d, J = 2.9 Hz, 1H), 4.64 - 4.59 (m, 1H), 4.58 - 4.39 (m, 11H), 4.36 (d, J = 7.8 Hz, 1H), 4.10 - 3.97 (m, 10H), 3.90 - 3.85 (m, 12H), 3.79 (d, J = 10.1 Hz, 1H), 3.62 (d, J = 8.8 Hz, 1H), 3.56 (s, 3H), 1.34 - 1.21 (m, 36H); 13¹³C NMR (101 MHz, D₂O) δ 103.75, 98.61, 98.54, 98.48, 97.65, 79.84, 79.55, 79.48, 79.41, 79.05, 78.34, 76.33, 76.26, 75.70, 75.54, 70.00, 69.04, 67.26, 67.19, 66.72, 66.64, 66.56, 57.29, 15.87, 15.69, 15.57; HRMS (ESI Orbitrap, negative) for C 73 H 111 Na 10 O 88 S 13 3- [M - 3Na] 3- m / z 1013.6524, found 1013.6528.

[0214] Compound 23

[0215] 1 ¹H NMR (400 MHz, D₂O) δ 5.41 (d, J = 3.8 Hz, 1H), 5.20 (s, 1H), 4.94 (d, J = 3.1 Hz, 1H), 4.76 - 4.63 (m, 3H), 4.53 (dd, J = 10.6, 3.8 Hz, 1H), 4.38 - 4.30 (m, 2H), 4.20 (t, J = 4.9 Hz, 1H), 3.44 (s, 3H), 1.47 (d, J = 6.6 Hz, 3H), 1.31 (d, J = 6.5 Hz, 3H); 13 ¹³C NMR (101 MHz, D₂O) δ 106.31, 96.95, 85.03, 84.66, 81.92, 79.06, 75.25, 72.32, 71.90, 66.60, 54.74, 16.79, 15.71; HRMS (ESI Orbitrap, negative) calcd for C 13 H 19 Na₄O 24 S₅ - [M - Na] - m / z 810.8466, found 810.8461.

[0216] Compound 24

[0217] 11H NMR (600 MHz, D2O) δ 5.44 (d, J = 3.8 Hz, 1H), 5.41 (d, J = 3.6 Hz, 1H), 5.23 (s, 1H), 4.97 - 4.94 (m, 3H), 4.77 (d, J = 1.7 Hz, 1H), 4.68 (qd, J = 6.5, 4.8 Hz, 1H), 4.57 (td, J = 9.8, 3.7 Hz, 2H), 4.52 (q, J = 6.7 Hz, 1H), 4.35 - 4.28 (m, 3H), 4.19 (t, J = 5.1 Hz, 1H), 3.46 (s, 3H), 1.50 (d, J = 6.5 Hz, 3H), 1.34 (d, J = 6.4 Hz, 3H), 1.31 (d, J = 6.5 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 106.35, 97.36, 97.33, 84.90, 84.85, 82.48, 79.77, 79.50, 75.48, 73.71, 73.63, 72.93, 72.34, 67.17, 66.94, 54.69, 16.87, 15.84, 15.79; HRMS (ESI Orbitrap, negative) calcd for C 19 H 32 O 34 S7 2- [M - 7Na + 5H] 2- m / z 513.9415, found 513.9415.

[0218] Compound 25

[0219] 1 1H NMR (600 MHz, D2O) δ 5.45 - 5.40 (m, 3H), 5.23 (s, 1H), 4.97 - 4.94 (m, 4H), 4.78 (d, J = 1.6 Hz, 1H), 4.69 (td, J = 6.5, 5.0 Hz, 1H), 4.60 - 4.54 (m, 4H), 4.47 - 4.40 (m, 2H), 4.34 - 4.29 (m, 3H), 4.20 (t, J = 5.1 Hz, 1H), 3.46 (s, 3H), 1.50 (d, J = 6.5 Hz, 3H), 1.37 - 1.32 (m, 9H); 1313C NMR (151 MHz, D2O) δ 106.35, 98.12, 97.39, 96.94, 84.90, 84.80, 82.73, 80.07, 79.78, 79.71, 75.56, 74.26, 74.17, 74.10, 73.72, 72.41, 72.27, 67.68, 67.23, 66.67, 54.68, 16.87, 16.01, 15.78, 15.75; HRMS (ESI Orbitrap, negative) calcd for C 25 H 41 O 44 S9 3- [M - 9Na + 6H] 3- m / z 444.2825, found 444.2828.

[0220] Compound 26

[0221] 1 1H NMR (600 MHz, D2O) δ 5.47 - 5.40 (m, 4H), 5.23 (s, 1H), 4.99 - 4.93 (m, 5H), 4.78 - 4.77 (m, 1H), 4.69 (qd, J = 6.5, 5.0 Hz, 1H), 4.60 - 4.55 (m, 4H), 4.52 (d, J = 6.5 Hz, 1H), 4.46 - 4.40 (m, 4H), 4.35 - 4.33 (m, 1H), 4.32 - 4.26 (m, 2H), 4.20 (t, J = 5.0 Hz, 1H), 3.46 (s, 3H), 1.50 (d, J = 6.5 Hz, 3H), 1.38 - 1.33 (m, 12H); 13 13C NMR (151 MHz, D2O) δ 106.38, 98.12, 97.40, 97.25, 84.94, 84.79, 82.67, 80.12, 79.77, 75.54, 74.86, 74.44, 74.09, 73.87, 73.77, 72.54, 72.30, 67.89, 67.36, 67.22, 66.77, 54.72, 16.88, 15.90, 15.83, 15.78; HRMS (ESI Orbitrap, negative) calcd for C 31 H 43 Na9O 54 S 11 2- [M - 2Na] 2- m / z 918.8318, found 918.8320.

[0222] Compound 27

[0223] 1 H NMR(600MHz,D2O)δ5.44 - 5.39(m,5H),5.22(s,1H),4.96 - 4.92(m,6H),4.77 - 4.76(m,1H),4.68 - 4.66(m,1H),4.55(dt,J=8.8,3.3Hz,5H),4.51(d,J=6.5Hz,1H),4.45 - 4.38(m,6H),4.33 - 4.31(m,1H),4.30 - 4.25(m,2H),4.19(t,J=5.1Hz,1H),3.45(s,3H),1.49(d,J=6.5Hz,3H),1.36 - 1.30(m,15H); 13 C NMR(151MHz,D2O)δ106.35,98.90,97.44,97.35,84.98,84.74,82.72,80.37,79.79,75.60,74.45,74.18,73.96,72.55,72.30,67.85,67.44,67.22,66.73,54.67,16.85,15.87,15.81,15.74;HRMS(ESI Orbitrap,negative)calcd for C 37 H 51 Na 11 O 64 S 13 2- [M - 2Na] 2- m / z 1093.7996,found 1093.8016.

[0224] Compound 28

[0225] 1 H NMR(400MHz,D2O)δ5.46 - 5.41(m,6H),5.23(s,1H),4.98 - 4.94(m,7H),4.68(s,1H),4.60 - 4.51(m,7H),4.48 - 4.26(m,11H),4.21(t,J=5.0Hz,1H),3.46(s,3H),1.50(d,J=6.5Hz,3H),1.40 - 1.30(m,18H); 1313C NMR (151 MHz, D2O) δ 106.36, 97.43, 84.95, 84.76, 82.69, 79.79, 75.58, 74.46, 74.14, 73.95, 72.55, 72.30, 67.22, 66.73, 54.68, 16.86, 15.88, 15.82, 15.75; HRMS (ESI Orbitrap, negative) calcd for C 43 H 59 Na 13 O 74 S 15 2- [M - 2Na] 2- m / z 1268.7673, found 1268.7684.

[0226] Compound 31

[0227] 1 1H NMR (400 MHz, D2O) δ 5.30 (d, J = 3.6 Hz, 1H), 4.75 (dd, J = 10.6, 3.2 Hz, 1H), 4.59 - 4.52 (m, 2H), 4.49 - 4.37 (m, 3H), 4.24 (d, J = 2.5 Hz, 2H), 3.90 (q, J = 6.5 Hz, 1H), 3.54 (s, 3H), 1.42 (d, J = 6.6 Hz, 3H), 1.29 (d, J = 6.5 Hz, 3H); 13 13C NMR (101 MHz, D2O) δ 102.20, 98.80, 78.74, 77.34, 75.86, 74.88, 72.83, 70.92, 70.79, 67.09, 57.15, 15.60, 15.17; HRMS (ESI Orbitrap, negative) calcd for C 13 H 20 Na3O 21 S4 - [M - Na] - m / z 708.9079, found 708.9074.

[0228] Compound 32

[0229] 11H NMR (600 MHz, D2O) δ 5.41 (d, J = 3.9 Hz, 1H), 5.32 (d, J = 3.6 Hz, 1H), 4.76 (dd, J = 10.5, 3.2 Hz, 1H), 4.62 - 4.58 (m, 4H), 4.47 - 4.41 (m, 3H), 4.29 - 4.19 (m, 3H), 4.15 (d, J = 3.1 Hz, 1H), 3.94 (q, J = 6.5 Hz, 1H), 3.57 (s, 3H), 1.46 (d, J = 6.5 Hz, 3H), 1.33 (d, J = 6.5 Hz, 3H), 1.27 (d, J = 6.6 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 102.11, 98.54, 94.75, 78.06, 77.35, 75.82, 75.01, 73.41, 73.40, 72.32, 70.92, 70.59, 69.44, 67.07, 66.55, 57.03, 15.68, 15.29, 15.23; HRMS (ESI Orbitrap, negative) calcd for C 19 H 29 Na4O 28 S5 - [M - Na] - m / z 956.9045, found 956.9039.

[0230] Compound 33

[0231] 1 1H NMR (400 MHz, D2O) δ 5.41 (d, J = 3.7 Hz, 1H), 5.31 (dd, J = 7.4, 3.7 Hz, 2H), 4.76 (d, J = 2.8 Hz, 2H), 4.67 (dd, J = 10.7, 3.7 Hz, 1H), 4.64 - 4.55 (m, 4H), 4.49 (q, J = 6.6 Hz, 1H), 4.45 - 4.36 (m, 3H), 4.30 - 4.19 (m, 4H), 4.13 (d, J = 2.5 Hz, 1H), 3.91 (q, J = 6.5 Hz, 1H), 3.54 (s, 3H); 1313C NMR (101 MHz, D2O) δ 102.12, 98.78, 98.68, 94.96, 79.67, 78.46, 77.56, 75.83, 74.89, 73.85, 73.58, 72.72, 72.41, 70.95, 70.80, 69.54, 67.74, 67.08, 56.99, 15.79, 15.54, 15.32, 15.21; HRMS (ESI Orbitrap, negative) calcd for C 25 H 37 Na5O 38 S7 2- [M - 2Na] 2- m / z 641.9254, found 641.9250.

[0232] Compound 34

[0233] 1 1H NMR (600 MHz, D2O) δ 5.43 (dd, J = 3.9, 2.0 Hz, 2H), 5.36 - 5.34 (m, 2H), 4.82 (d, J = 2.7 Hz, 1H), 4.78 - 4.75 (m, 1H), 4.70 - 4.57 (m, 7H), 4.48 - 4.39 (m, 4H), 4.32 (d, J = 2.8 Hz, 1H), 4.30 - 4.22 (m, 4H), 4.15 (d, J = 3.2 Hz, 2H), 3.94 (q, J = 6.5 Hz, 1H), 3.57 (s, 3H), 1.47 (d, J = 6.4 Hz, 3H), 1.42 (d, J = 6.6 Hz, 3H), 1.35 - 1.32 (m, 6H), 1.28 (d, J = 6.6 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 102.07, 98.71, 98.65, 94.86, 94.59, 79.43, 78.45, 77.57, 75.80, 75.03, 73.92, 73.57, 73.45, 73.43, 73.20, 72.42, 72.35, 70.93, 70.59, 69.46, 69.37, 67.83, 67.09, 66.56, 56.94, 15.78, 15.63, 15.31, 15.23; HRMS (ESI Orbitrap, negative) calcd for C 31 H 46 Na6O 45 S8 2- [M - 2Na] 2- m / z 765.9237, found 765.9236.

[0234] Compound 35

[0235] 1 H NMR(600MHz,D2O)δ5.45 - 5.43(m,2H),5.37 - 5.33(m,3H),4.83 - 4.80(m,3H),4.73 - 4.57(m,8H),4.56 - 4.51(m,1H),4.48 - 4.38(m,4H),4.33 - 4.23(m,6H),4.18 - 4.13(m,2H),3.94(q,J=6.5Hz,1H),3.57(s,3H),1.46(d,J=6.4Hz,3H),1.42(dd,J=6.7,4.6Hz,6H),1.36 - 1.31(m,9H); 13 C NMR(151MHz,D2O)δ102.12,98.71,98.63,94.94,94.81,79.52,79.44,78.50,77.62,75.84,74.86,73.90,73.80,73.63,73.60,73.41,72.65,72.42,72.40,70.93,70.78,69.50,69.43,67.80,67.78,67.07,66.92,57.05,15.80,15.62,15.52,15.33,15.32,15.20;HRMS(ESI Orbitrap,negative)calcd for C 37 H 54 Na8O 55 S 10 2- [M - 2Na] 2- m / z 940.8914,found 940.8918.

[0236] Compound 36

[0237] 1 H - NMR(600MHz,D2O)δ5.45 - 5.28(m,6H),4.82 - 4.79(m,2H),4.73(dd,J=10.5,3.3Hz,1H),4.68 - 4.54(m,10H),4.48 - 4.33(m,5H),4.32 - 4.19(m,7H),4.13(d,J=3.0Hz,3H),3.91(q,J=6.5Hz,1H),3.55(s,3H),1.46 - 1.36(m,9H),1.30(dd,J=6.6,3.3Hz,9H),1.26(d,J=6.5Hz,3H);13 C NMR(151MHz, D2O) δ 102.10, 98.66, 98.64, 98.61, 94.77, 94.73, 94.58, 79.43, 79.30, 78.52, 77.67, 75.86, 75.02, 73.94, 73.90, 73.67, 73.46, 73.40, 73.36, 73.21, 72.45, 72.40, 72.35, 70.91, 70.57, 69.40, 69.36, 67.87, 67.78, 67.09, 67.04, 66.90, 66.57, 57.04, 15.82, 15.63, 15.33, 15.31, 15.23; HRMS(ESI Orbitrap, negative) calcd for C 43 H 63 Na9O 62 S 11 2- [M - 2Na] 2- m / z 1064.8897, found 1064.8902.

[0238] Compound 38

[0239] 1 H NMR(600MHz, D2O) δ 5.47 - 5.42(m, 4H), 5.38(d, J = 3.7Hz, 1H), 5.36 - 5.30(m, 3H), 4.87 - 4.83(m, 3H), 4.76(d, J = 3.4Hz, 1H), 4.71 - 4.69(m, 3H), 4.66 - 4.58(m, 10H), 4.52 - 4.49(m, 1H), 4.48 - 4.43(m, 4H), 4.42 - 4.39(m, 1H), 4.33 - 4.31(m, 3H), 4.29(d, J = 2.4Hz, 1H), 4.27 - 4.24(m, 5H), 4.17 - 4.15(m, 4H), 3.94(q, J = 6.5Hz, 1H), 3.58(s, 3H), 1.47 - 1.41(m, 12H), 1.33(t, J = 5.7Hz, 12H), 1.29(d, J = 6.7Hz, 3H); 1313C NMR(151MHz,D2O)δ102.13,98.62,98.59,98.49,94.68,94.59,94.36,79.30,79.23,78.88,78.47,77.69,75.87,75.04,73.92,73.90,73.72,73.68,73.46,73.40,73.34,73.26,73.23,73.20,72.96,72.49,72.42,72.41,72.38,70.95,70.61,69.37,69.18,67.94,67.84,67.78,67.12,67.07,66.84,66.80,66.62,57.11,15.85,15.66,15.63,15.57,15.40,15.35,15.27;HRMS(ESI Orbitrap,negative)calcd for C 55 H 80 Na 11 O 79 S 14 3- [M - 3Na] 3- m / z901.5741,found901.5761.

[0240] Compound 39

[0241] 1 1H NMR(600MHz,D2O)δ5.43 - 5.31(m,4H),5.30 - 5.20(m,5H),4.78 - 4.73(m,3H),4.69 - 4.68(m,2H),4.63 - 4.58(m,4H),4.56 - 4.48(m,9H),4.47 - 4.27(m,8H),4.25 - 4.12(m,10H),4.07(d,J=3.6Hz,4H),3.85(q,J=6.2Hz,1H),3.48(s,3H),1.38 - 1.31(m,15H),1.25 - 1.22(m,15H); 1313C NMR(151MHz,D2O)δ102.12,98.85,98.73,98.61,98.52,98.39,95.09,94.71,94.48,94.25,79.55,79.28,78.90,78.84,78.56,78.53,78.53,77.68,77.68,77.35,75.87,74.87,73.87,73.81,73.79,73.69,73.65,73.59,73.43,73.30,73.24,73.19,73.18,73.16,72.85,72.67,72.43,72.41,70.93,70.80,69.57,69.38,69.26,69.13,67.92,67.87,67.78,67.08,66.94,66.79,66.65,57.10,57.08,15.83,15.61,15.57,15.54,15.52,15.40,15.38,15.36,15.33,15.20;HRMS(ESI Orbitrap,negative)calcd for C 61 H 88 Na 13 O 89 S 16 3- [M - 3Na] 3- m / z 1018.2193,found 1018.2201.

[0242] Compound 40

[0243] 1 1H NMR(600MHz,D2O)δ5.38 - 5.28(m,6H),5.24 - 5.19(m,4H),4.69 - 4.64(m,5H),4.62 - 4.47(m,16H),4.43 - 4.28(m,7H),4.25 - 4.09(m,11H),4.05(q,J=3.8Hz,5H),3.84(q,J=6.0Hz,1H),3.47(s,3H),1.39 - 1.29(m,15H),1.27 - 1.22(m,15H),1.18(d,J=6.6Hz,3H); 1313C NMR(151MHz,D2O)δ102.13,98.62,98.41,98.34,94.69,94.63,94.17,94.10,79.28,78.66,78.57,78.51,77.74,75.89,75.04,74.88,73.86,73.74,73.58,73.43,73.38,73.27,73.17,72.81,72.68,72.47,72.38,70.93,70.60,69.56,69.41,69.36,69.09,69.04,67.95,67.86,67.75,67.11,67.02,66.77,66.70,66.60,57.06,15.86,15.62,15.55,15.42,15.39,15.33,15.25; HRMS(ESI Orbitrap, negative) calcd for C 67 H 97 Na 13 O 96 S 17 4- [M - 4Na] 4- m / z 819.9163, found 819.9168.

[0244] Compound 41

[0245] 1 1H NMR(600MHz,D2O)δ5.47 - 5.42(m,5H),5.37(d,J = 3.8Hz,1H),5.34 - 5.25(m,5H),4.88 - 4.81(m,6H),4.71 - 4.56(m,17H),4.54 - 4.47(m,4H),4.46 - 4.35(m,4H),4.33 - 4.20(m,12H),4.14(d,J = 3.3Hz,5H),3.92(q,J = 6.5Hz,1H),3.56(s,3H),1.46 - 1.38(m,18H),1.33 - 1.30(m,18H); 1313C NMR(151MHz,D2O)δ102.03,98.62,98.51,98.41,98.23,94.98,94.61,94.36,94.08,94.05,79.45,79.16,78.79,78.46,78.40,77.61,75.78,74.77,73.77,73.71,73.69,73.62,73.53,73.47,73.43,73.32,73.19,73.14,73.06,72.68,72.57,72.33,72.28,70.83,70.70,69.47,69.27,69.15,68.99,67.82,67.76,67.72,67.66,67.00,66.97,66.68,66.52,56.96,15.75,15.51,15.46,15.44,15.31,15.28,15.26,15.23,15.10; HRMS(ESI Orbitrap,negative) calcd for C 73 H 105 Na 15 O 106 S 19 4- [M - 4Na] 4- m / z 907.4002, found 907.4010.

[0246] Compound 42

[0247] 1 1H NMR(600MHz,D2O)δ5.10(d,J = 3.9Hz,1H),4.92(d,J = 3.0Hz,1H),4.66(dd,J = 10.7,3.0Hz,1H),4.61(q,J = 6.6Hz,1H),4.46(d,J = 7.8Hz,1H),4.37(dd,J = 10.2,3.0Hz,1H),4.19(d,J = 3.0Hz,1H),4.00(dd,J = 10.7,3.9Hz,1H),3.92(q,J = 6.6Hz,1H),3.75(dd,J = 10.2,7.8Hz,1H),3.58(s,3H),1.40(d,J = 6.6Hz,3H),1.35(d,J = 6.5Hz,3H); 13CNMR(151MHz, D2O) δ 103.27, 99.99, 79.45, 79.13, 76.85, 74.89, 70.96, 68.76, 66.74, 66.71, 57.13, 15.94, 15.43; HRMS(ESI Orbitrap, negative) calcd for C 13 H 21 Na2O 18 S3 - [M - Na] - m / z 606.9691, found 606.9687.

[0248] Compound 43

[0249] 1 1H NMR(600MHz, D2O) δ 5.15(d, J = 3.9Hz, 1H), 5.06(d, J = 4.0Hz, 1H), 4.90(d, J = 3.0Hz, 1H), 4.79 - 4.78(m, 1H), 4.61(dd, J = 10.6, 3.1Hz, 1H), 4.56(q, J = 6.5Hz, 1H), 4.49 - 4.45(m, 2H), 4.36(dd, J = 10.3, 3.0Hz, 1H), 4.17(d, J = 3.1Hz, 1H), 4.06(dd, J = 10.7, 2.9Hz, 1H), 3.97(dd, J = 10.6, 3.9Hz, 1H), 3.94 - 3.87(m, 2H), 3.68(dd, J = 10.3, 7.8Hz, 1H), 3.56(s, 3H), 1.40(d, J = 6.6Hz, 3H), 1.33(d, J = 6.4Hz, 3H), 1.26(d, J = 6.5Hz, 3H); 13 CNMR(151MHz, D2O) δ 103.11, 100.33, 99.02, 79.99, 79.35, 79.06, 76.70, 76.59, 75.37, 71.03, 68.83, 67.25, 66.64, 66.58, 66.45, 56.94, 15.87, 15.66, 15.31; HRMS(ESI Orbitrap, negative) calcd for C 19 H 30 Na3O 25 S4 - [M - Na] - m / z 854.9658, found 854.9651.

[0250] Compound 44

[0251] 1 1H NMR (600 MHz, D2O) δ 5.19 (d, J = 4.0 Hz, 1H), 5.11 (d, J = 3.9 Hz, 1H), 5.07 (d, J = 4.0 Hz, 1H), 4.92 (d, J = 3.1 Hz, 1H), 4.80 (s, 1H), 4.68 (dd, J = 10.7, 3.0 Hz, 1H), 4.66 - 4.59 (m, 2H), 4.56 (q, J = 6.5 Hz, 1H), 4.46 (d, J = 7.8 Hz, 1H), 4.41 (q, J = 6.7 Hz, 1H), 4.36 (dd, J = 10.2, 3.0 Hz, 1H), 4.24 (d, J = 3.1 Hz, 1H), 4.18 (d, J = 3.1 Hz, 1H), 4.08 (dd, J = 10.7, 3.0 Hz, 1H), 4.04 - 3.96 (m, 3H), 3.92 (q, J = 6.6 Hz, 1H), 3.69 (dd, J = 10.2, 7.8 Hz, 1H), 3.57 (s, 3H), 1.40 (d, J = 6.6 Hz, 3H), 1.34 (m, 9H); 13 13C NMR (151 MHz, D2O) δ 103.15, 100.37, 99.91, 99.13, 79.96, 79.53, 79.08, 77.90, 76.76, 76.50, 76.46, 74.95, 71.06, 68.84, 67.76, 67.30, 66.77, 66.73, 66.67, 66.61, 56.97, 15.98, 15.91, 15.34, 15.22; HRMS (ESI Orbitrap, negative) calcd for C 25 H 39 Na3O 32 S5 2- [M - 2Na] 2- m / z 539.9866, found 539.9862.

[0252] Compound 45

[0253] 11H NMR (400 MHz, D2O) δ 5.17 - 5.14 (m, 2H), 5.05 (dd, J = 8.9, 4.0 Hz, 2H), 4.88 (d, J = 3.1 Hz, 1H), 4.76 - 4.75 (m, 2H), 4.63 - 4.52 (m, 4H), 4.48 - 4.40 (m, 3H), 4.34 (dd, J = 10.2, 3.0 Hz, 1H), 4.21 (d, J = 3.0 Hz, 1H), 4.15 (d, J = 3.1 Hz, 1H), 4.07 (d, J = 10.8 Hz, 2H), 3.99 - 3.85 (m, 5H), 3.67 (dd, J = 10.3, 7.7 Hz, 1H), 3.54 (s, 3H), 1.38 (d, J = 6.6 Hz, 3H), 1.32 - 1.30 (m, 9H), 1.25 (d, J = 6.4 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 103.92, 101.21, 101.08, 100.04, 99.83, 80.85, 80.76, 80.13, 79.86, 78.58, 77.65, 77.54, 77.44, 77.14, 76.16, 71.82, 69.60, 68.73, 68.12, 68.10, 67.54, 67.42, 67.39, 67.36, 67.31, 57.71, 16.70, 16.67, 16.48, 16.08, 15.86; HRMS (ESI Orbitrap, negative) calcd for C 31 H 48 Na4O 39 S6 2- [M - 2Na] 2- m / z 663.9849, found 663.9846.

[0254] Compound 46

[0255] 11H NMR (600 MHz, D2O) δ 5.20 (dd, J = 7.1, 4.0 Hz, 2H), 5.11 (d, J = 3.9 Hz, 1H), 5.09 - 5.06 (m, 2H), 4.93 - 4.91 (m, 1H), 4.80 - 4.79 (m, 2H), 4.68 (dd, J = 10.7, 3.0 Hz, 1H), 4.65 - 4.60 (m, 3H), 4.57 (dt, J = 12.4, 5.6 Hz, 2H), 4.47 - 4.40 (m, 3H), 4.36 (dd, J = 10.3, 3.0 Hz, 1H), 4.23 (dd, J = 5.5, 3.1 Hz, 2H), 4.17 (d, J = 3.1 Hz, 1H), 4.09 (dt, J = 10.7, 3.4 Hz, 2H), 4.05 - 3.90 (m, 6H), 3.69 (dd, J = 10.2, 7.8 Hz, 1H), 3.56 (s, 3H), 1.40 (d, J = 6.6 Hz, 3H), 1.35 - 1.33 (m, 15H); 13 13C NMR (151 MHz, D2O) δ 103.89, 101.19, 101.08, 100.63, 100.02, 99.86, 80.75, 80.29, 79.83, 78.61, 78.58, 77.63, 77.53, 77.22, 77.11, 75.69, 71.79, 69.57, 68.71, 68.57, 68.11, 68.08, 67.50, 67.47, 67.40, 67.37, 67.33, 57.69, 16.73, 16.69, 16.64, 16.05, 15.99, 15.82; HRMS (ESI Orbitrap, negative) calcd for C 37 H 57 Na5O 46 S7 2- [M - 2Na] 2- m / z 787.9833, found 787.9833.

[0256] Compound 47

[0257] 11H NMR (600 MHz, D2O) δ 5.23 - 5.16 (m, 3H), 5.11 - 5.04 (m, 3H), 4.91 (d, J = 3.1 Hz, 1H), 4.86 - 4.80 (m, 3H), 4.66 - 4.53 (m, 6H), 4.52 - 4.41 (m, 4H), 4.36 (dd, J = 10.2, 2.9 Hz, 1H), 4.24 (dd, J = 8.3, 3.1 Hz, 2H), 4.17 (d, J = 3.1 Hz, 1H), 4.12 - 4.08 (m, 3H), 4.00 - 3.89 (m, 7H), 3.70 (dd, J = 10.2, 7.7 Hz, 1H), 3.57 (s, 3H), 1.40 (d, J = 6.5 Hz, 3H), 1.35 - 1.33 (m, 15H), 1.28 (d, J = 6.4 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 103.89, 101.20, 101.09, 100.90, 100.06, 99.73, 99.72, 80.79, 80.71, 80.07, 79.83, 78.66, 78.27, 77.66, 77.57, 77.34, 77.19, 77.11, 77.04, 76.13, 71.78, 69.57, 68.77, 68.71, 68.12, 68.10, 68.06, 67.51, 67.46, 67.39, 67.36, 67.33, 67.29, 57.70, 16.70, 16.64, 16.48, 16.05, 15.91, 15.80; HRMS (ESI Orbitrap, negative) calcd for C 43 H 66 Na6O 53 S8 2- [M - 2Na] 2- m / z 911.9816, found 911.9815.

[0258] Compound 48

[0259] 11H NMR (600 MHz, D2O) δ 5.25 - 5.15 (m, 3H), 5.12 - 5.05 (m, 4H), 4.94 - 4.90 (m, 1H), 4.80 - 4.79 (m, 3H), 4.69 - 4.53 (m, 8H), 4.48 - 4.41 (m, 4H), 4.36 (dd, J = 10.3, 3.0 Hz, 1H), 4.27 - 4.20 (m, 3H), 4.17 (d, J = 3.1 Hz, 1H), 4.12 - 4.08 (m, 3H), 4.05 - 3.89 (m, 8H), 3.69 (dd, J = 10.2, 7.7 Hz, 1H), 3.56 (s, 3H), 1.40 (d, J = 6.6 Hz, 3H), 1.35 - 1.33 (m, 21H); 13 13C NMR (151 MHz, D2O) δ 103.92, 101.24, 101.12, 100.91, 100.65, 100.09, 99.77, 99.67, 80.80, 80.76, 80.70, 80.31, 79.86, 78.71, 78.56, 78.28, 77.71, 77.60, 77.23, 77.13, 77.11, 77.03, 75.72, 71.81, 69.59, 68.80, 68.73, 68.61, 68.15, 68.13, 68.08, 67.53, 67.51, 67.46, 67.41, 67.39, 67.36, 57.72, 16.76, 16.73, 16.66, 16.07, 16.04, 15.93, 15.83; HRMS (ESI Orbitrap, negative) calcd for C 49 H 75 Na7O 60 S9 2- [M - 2Na] 2- m / z 1035.9799, found 1035.9800.

[0260] Compound 49

[0261] 11H NMR (600 MHz, D2O) δ 5.25 - 5.15 (m, 4H), 5.11 - 5.04 (m, 4H), 4.91 (d, J = 3.1 Hz, 1H), 4.80 - 4.79 (m, 4H), 4.66 - 4.52 (m, 8H), 4.52 - 4.41 (m, 5H), 4.36 (dd, J = 10.3, 3.0 Hz, 1H), 4.24 (dd, J = 10.6, 3.1 Hz, 3H), 4.17 (d, J = 3.0 Hz, 1H), 4.12 - 4.07 (m, 4H), 4.01 - 3.88 (m, 9H), 3.69 (dd, J = 10.2, 7.7 Hz, 1H), 3.56 (s, 3H), 1.40 (d, J = 6.6 Hz, 3H), 1.35 - 1.33 (m, 21H), 1.28 (d, J = 6.5 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 103.13, 100.46, 100.31, 100.06, 100.02, 99.30, 98.96, 98.71, 98.60, 80.04, 80.00, 79.85, 79.82, 79.32, 79.07, 77.93, 77.37, 77.35, 76.93, 76.81, 76.59, 76.33, 76.23, 76.18, 76.12, 76.03, 75.37, 71.01, 68.79, 68.03, 68.00, 67.92, 67.35, 67.25, 66.74, 66.63, 66.59, 66.56, 66.52, 56.92, 15.99, 15.94, 15.86, 15.71, 15.27, 15.19, 15.15, 15.04; HRMS (ESI Orbitrap, negative) calcd for C 55 H 84 Na8O 67 S 10 2- [M - 2Na] 2- m / z 1159.9783, found 1159.9788.

[0262] Compound 50

[0263] 11H NMR (600 MHz, D2O) δ 5.25 - 5.18 (m, 4H), 5.13 - 5.06 (m, 5H), 4.93 (d, J = 3.0 Hz, 1H), 4.81 - 4.80 (m, 4H), 4.68 (dd, J = 10.7, 3.0 Hz, 1H), 4.66 - 4.55 (m, 9H), 4.49 - 4.41 (m, 5H), 4.37 (dd, J = 10.3, 3.0 Hz, 1H), 4.28 - 4.21 (m, 4H), 4.18 (d, J = 3.1 Hz, 1H), 4.13 - 4.09 (m, 4H), 4.06 - 3.91 (m, 10H), 3.70 (dd, J = 10.2, 7.7 Hz, 1H), 3.57 (s, 3H), 1.41 (d, J = 6.6 Hz, 3H), 1.38 - 1.32 (m, 27H); 13 13C NMR (151 MHz, D2O) δ 103.16, 100.44, 100.32, 100.12, 100.09, 99.89, 99.26, 99.01, 98.81, 98.68, 80.02, 79.99, 79.90, 79.87, 79.52, 79.09, 77.90, 77.82, 77.45, 77.43, 76.90, 76.76, 76.48, 76.37, 76.27, 76.23, 76.10, 74.95, 71.05, 68.85, 68.07, 68.03, 67.97, 67.85, 67.35, 67.27, 66.75, 66.75, 66.73, 66.68, 66.66, 66.63, 66.59, 56.97, 16.02, 16.00, 15.97, 15.91, 15.32, 15.28, 15.21, 15.18, 15.09; HRMS (ESI Orbitrap, negative) calcd for C 61 H 93 Na9O 74 S 11 2- [M - 2Na] 2- m / z 1283.9766, found 1283.9773.

[0264] Compound 51

[0265] 11H NMR (600 MHz, D2O) δ 5.27 - 5.15 (m, 5H), 5.13 - 5.04 (m, 5H), 4.92 (d, J = 3.2 Hz, 1H), 4.86 - 4.81 (m, 5H), 4.67 - 4.55 (m, 10H), 4.52 - 4.41 (m, 6H), 4.37 (dd, J = 10.3, 2.9 Hz, 1H), 4.27 - 4.21 (m, 4H), 4.17 (d, J = 3.1 Hz, 1H), 4.10 (m, 5H), 4.03 - 3.88 (m, 11H), 3.70 (dd, J = 10.2, 7.8 Hz, 1H), 3.57 (s, 3H), 1.41 (d, J = 6.6 Hz, 3H), 1.37 - 1.31 (m, 27H), 1.28 (d, J = 6.5 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 103.16, 100.46, 100.32, 100.09, 100.05, 100.04, 99.27, 98.94, 98.76, 98.66, 98.46, 80.05, 80.02, 79.87, 79.79, 79.53, 79.33, 79.09, 77.91, 77.41, 77.40, 77.30, 76.92, 76.78, 76.56, 76.37, 76.28, 76.21, 76.17, 75.85, 75.40, 71.05, 68.84, 68.06, 68.02, 67.96, 67.36, 67.27, 67.24, 66.76, 66.67, 66.64, 66.59, 66.55, 56.96, 16.03, 15.98, 15.90, 15.75, 15.32, 15.23, 15.19, 15.08; HRMS (ESI Orbitrap, negative) calcd for C 67 H 102 Na9O 81 S 12 3- [M - 3Na] 3- m / z 930.9869, found 930.9881.

[0266] Compound 52

[0267] 11H NMR (600 MHz, D2O) δ 5.24 - 5.20 (m, 5H), 5.14 - 5.05 (m, 6H), 4.93 (d, J = 3.1 Hz, 1H), 4.83 - 4.80 (m, 5H), 4.66 - 4.55 (m, 12H), 4.48 - 4.42 (m, 6H), 4.37 (dd, J = 10.3, 3.0 Hz, 1H), 4.25 (dd, J = 6.2, 3.1 Hz, 5H), 4.18 (d, J = 3.1 Hz, 1H), 4.13 - 4.09 (m, 5H), 4.06 - 3.90 (m, 12H), 3.70 (dd, J = 10.2, 7.7 Hz, 1H), 3.57 (s, 3H), 1.41 (d, J = 6.6 Hz, 3H), 1.36 - 1.34 (m, 33H); 13 13C NMR (151 MHz, D2O) δ 103.16, 100.46, 100.32, 100.11, 100.05, 100.02, 99.89, 99.28, 99.01, 98.76, 98.62, 98.47, 80.02, 79.99, 79.87, 79.85, 79.79, 79.54, 79.10, 77.91, 77.82, 77.42, 77.30, 76.92, 76.78, 76.48, 76.37, 76.27, 76.22, 76.17, 76.04, 75.87, 74.96, 71.05, 68.84, 68.06, 68.02, 67.97, 67.85, 67.36, 67.27, 67.27, 67.23, 66.74, 66.69, 66.66, 66.63, 66.59, 56.96, 16.03, 16.00, 15.97, 15.91, 15.32, 15.28, 15.24, 15.22, 15.19, 15.09; HRMS (ESI Orbitrap, negative) calcd for C 73 H 111 Na 10 O 88 S 13 3- [M - 3Na] 3- m / z 1013.6524, found 1013.6536.

[0268] Compound 53

[0269] 11H NMR (600 MHz, D2O) δ 5.33 (d, J = 3.6 Hz, 1H), 4.99 (d, J = 3.1 Hz, 1H), 4.84 (dd, J = 10.7, 3.1 Hz, 1H), 4.60 - 4.54 (m, 3H), 4.48 - 4.39 (m, 2H), 4.27 (d, J = 2.5 Hz, 1H), 3.92 (q, J = 6.5 Hz, 1H), 3.57 (d, J = 0.8 Hz, 3H), 1.45 (d, J = 6.5 Hz, 3H), 1.36 (d, J = 6.4 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 102.17, 98.74, 79.72, 78.91, 77.24, 75.82, 72.68, 72.25, 70.86, 66.90, 57.11, 15.70, 15.52; HRMS (ESI Orbitrap, negative) calcd for C 13 H 19 Na4O 24 S5 - [M - Na] - m / z 810.8466, found 810.8469.

[0270] Compound 54

[0271] 1 1H NMR (400 MHz, D2O) δ 5.40 (d, J = 3.7 Hz, 1H), 5.35 (d, J = 3.5 Hz, 1H), 4.93 (d, J = 6.0 Hz, 3H), 4.61 - 4.50 (m, 4H), 4.47 (t, J = 6.4 Hz, 1H), 4.42 - 4.33 (m, 3H), 4.24 (s, 1H), 3.89 (q, J = 6.5 Hz, 1H), 3.51 (s, 3H), 1.40 (d, J = 6.5 Hz, 3H), 1.33 (d, J = 6.4 Hz, 3H), 1.28 (d, J = 6.5 Hz, 3H); 13 13C NMR (101 MHz, D2O) δ 102.19, 98.47, 96.30, 79.61, 79.54, 78.40, 77.55, 75.84, 74.04, 72.45, 72.37, 72.31, 70.93, 67.48, 66.68, 56.98, 16.00, 15.92, 15.85; HRMS (ESI Orbitrap, negative) calcd for C 19 H 27 Na5O 34 S7 2-[M - 2Na] 2- m / z 568.8964, found 568.8955.

[0272] Compound 55

[0273] 1 H NMR (400 MHz, D2O) δ 5.45 (d, J = 3.6 Hz, 1H), 5.38 (d, J = 3.6 Hz, 1H), 5.31 (d, J = 3.6 Hz, 1H), 4.97 (dd, J = 7.6, 2.8 Hz, 2H), 4.90 (dd, J = 10.8, 2.8 Hz, 1H), 4.84 (d, J = 8.1 Hz, 1H), 4.74 - 4.70 (m, 1H), 4.63 - 4.52 (m, 5H), 4.48 - 4.36 (m, 4H), 4.28 (dd, J = 12.4, 2.5 Hz, 2H), 3.91 (q, J = 6.4 Hz, 1H), 3.54 (s, 3H), 1.44 (d, J = 6.5 Hz, 3H), 1.40 (d, J = 6.6 Hz, 3H), 1.35 (t, J = 7.3 Hz, 6H); 13 C NMR (101 MHz, D2O) δ 102.11, 98.66, 98.56, 96.43, 79.90, 79.78, 79.67, 78.60, 77.57, 75.79, 73.88, 73.41, 72.62, 72.37, 72.29, 70.90, 67.93, 67.49, 66.92, 56.94, 15.98, 15.86, 15.78, 15.55; HRMS (ESI Orbitrap, negative) calcd for C 25 H 35 Na7O 44 S9 2- [M - 2Na] 2- m / z 743.8641, found 743.8641.

[0274] Compound 56

[0275] 11H NMR (600 MHz, D2O) δ 5.47 - 5.35 (m, 4H), 4.99 - 4.91 (m, 5H), 4.69 - 4.67 (m, 1H), 4.62 - 4.52 (m, 6H), 4.46 - 4.36 (m, 6H), 4.32 - 4.27 (m, 2H), 3.93 (q, J = 6.4 Hz, 1H), 3.55 (s, 3H), 1.45 (d, J = 6.4 Hz, 3H), 1.41 (d, J = 6.6 Hz, 3H), 1.37 (t, J = 6.5 Hz, 6H), 1.32 (d, J = 6.5 Hz, 3H); 13 13C NMR (151 MHz, D2O) δ 102.10, 98.77, 98.57, 96.42, 79.97, 79.66, 79.59, 78.79, 77.69, 75.82, 74.05, 73.97, 73.67, 72.53, 72.41, 72.30, 72.25, 70.91, 68.02, 67.51, 67.45, 66.68, 56.91, 15.99, 15.93, 15.85, 15.79; HRMS (ESI Orbitrap, negative) calcd for C 31 H 43 Na9O 54 S 11 2- [M - 2Na] 2- m / z 918.8318, found 918.8321.

[0276] Compound 57

[0277] 1 1H NMR (600 MHz, D2O) δ 5.49 - 5.43 (m, 3H), 5.39 (d, J = 3.7 Hz, 1H), 5.34 (d, J = 3.7 Hz, 1H), 5.00 - 4.97 (m, 3H), 4.93 (td, J = 8.1, 4.1 Hz, 2H), 4.86 (dd, J = 10.7, 3.0 Hz, 1H), 4.74 (dd, J = 10.8, 3.5 Hz, 1H), 4.68 (dd, J = 10.7, 3.5 Hz, 1H), 4.64 - 4.60 (m, 3H), 4.57 - 4.50 (m, 4H), 4.47 - 4.36 (m, 6H), 4.31 (dd, J = 11.1, 2.6 Hz, 3H), 3.93 (q, J = 6.5 Hz, 1H), 3.56 (s, 3H), 1.46 (d, J = 6.4 Hz, 3H), 1.42 (dd, J = 6.7, 2.3 Hz, 6H), 1.39 - 1.36 (m, 9H); 1313C NMR (151 MHz, D2O) δ 102.15, 98.76, 98.63, 98.57, 96.84, 96.52, 80.02, 79.84, 79.82, 79.69, 78.87, 77.79, 75.85, 74.14, 73.98, 73.66, 73.44, 72.85, 72.61, 72.53, 72.41, 72.30, 70.92, 68.02, 67.52, 67.42, 66.92, 56.99, 16.00, 15.97, 15.82, 15.53; HRMS (ESI Orbitrap, negative) calcd for C 37 H 51 Na 11 O 64 S 13 2- [M - 2Na] 2- m / z 1093.7996, found 1093.7999.

[0278] The present invention does not elaborate on the preparation methods of the compounds in Example 5, and they can be prepared by using preparation methods similar to those in Examples 1 - 4 of the present invention.

[0279] Anticoagulant Activity Test

[0280] 1) Experimental instruments: Hemagglutination analyzer (model MC - 4000, TECO GmbH, Germany); Vortex oscillator (model Vortex Genie, Scientific Industrie, USA); Electronic balance (model ME104E, Mettler Toledo, Switzerland).

[0281] 2) Experimental materials: Human coagulation control plasma (Coagulation Control Plasma), Lot 093B - N103A; APTT reagent, Lot 20003262, Lot 20003461; CaCl2 solution, Lot 10353256, Lot 210123; PT reagent, Lot 10003512; TT reagent, Lot 30003454, Lot 30003400; MDC Hemostasis, product of TECO GmbH (Germany); Enoxaparin sodium injection (Enoxaparin, Enox), 0.4 mL: 4000 AXa IU, Lot AS735A, Sanofi (Beijing) Pharmaceutical Co., Ltd.

[0282] 3) Experimental method:

[0283] ① Solution preparation: Coagulation quality control plasma, PT reagent, and TT reagent solution.

[0284] Sample solution: Weigh an appropriate amount of the compound, dissolve it in pure water to prepare a stock solution of 12.8 mg / mL, and then dilute it with pure water to a 1.28 mg / mL solution. For active compounds, continue to dilute it in a 2-fold gradient to prepare solutions with lower concentrations.

[0285] Enoxaparin solution: Pipette 12.8 μL of enoxaparin sodium injection (100 mg / mL), add 987.2 μL of pure water to dilute it to 1280 μg / mL, and then dilute it with pure water to the required concentration.

[0286] ② Activated partial thromboplastin time (APTT) test: Add 45 μL of human coagulation quality control plasma and 5 μL of the sample solution or solvent to a test tube pre-warmed at 37°C, and incubate at 37°C for 2 min; Pipette 50 μL of APTT reagent pre-warmed at 37°C into the test tube and incubate at 37°C for 3 min; Transfer the test tube to the detection well, add 50 μL of CaCl2 solution pre-warmed at 37°C, start timing, and record the clotting time.

[0287] ③ Prothrombin time (PT) test: Add 45 μL of human coagulation quality control plasma and 5 μL of the sample solution or solvent to a test tube pre-warmed at 37°C, and incubate at 37°C for 2 min; Transfer the test tube to the detection well, add 100 μL of PT reagent pre-warmed at 37°C, start timing, and record the clotting time.

[0288] ④ Thrombin time (TT) test: Add 90 μL of human coagulation quality control plasma and 10 μL of the sample solution or solvent to a test tube pre-warmed at 37°C, and incubate at 37°C for 2 min; Transfer the test tube to the detection well, add 50 μL of TT reagent pre-warmed at 37°C, start timing, and record the clotting time.

[0289] 4) Data processing: Input the experimental data into an Excel document and save it. Use GraphPad Prism 8.0 to perform linear fitting on the compound concentration - mean clotting time, and calculate the drug concentration that doubles the clotting time (EC 2.0× ).

[0290] The results of the APTT experiment are shown in Table 1, indicating that at a concentration of 128 μg / mL, compounds 25, 26, 27, 28, 29, 30, 35, 36, 37, 38, 39, 40, 41, 56, 57, and 58 can extend the human plasma APTT to 2 times or more of the control group. Among them, compounds 28, 29, 30, 37, and 58 have strong activities and can extend the APTT to more than 12 times of the control group.

[0291] Table 1 Effects of fucoidan sulfate compounds on APTT of human coagulation quality control plasma (mean±SD, n = 3)

[0292]

[0293]

[0294] Furthermore, a series of concentrations of the active compounds were detected, and the compound concentrations required to double APTT were calculated (EC 2.0× )(as shown in Table 2). It can be seen that compounds 29, 30, 37, and 58 have strong activities, and the EC 2.0× are 13.6 μg / mL, 14.5 μg / mL, 16.4 μg / mL, and 12.9 μg / mL, respectively. The EC 2.0× of the positive control enoxaparin is 7.0 μg / mL.

[0295] Table 2 Plasma APTT prolonging activities of some fucoidan sulfate compounds

[0296] Compound Concentration - APTT Fitting Equation <![CDATA[R 2 > <![CDATA[EC 2.0× (μg / mL)]]> 25 y = 0.3786x + 37.58 0.997 65.6 26 y = 0.7922x + 36.52 0.990 32.7 27 y = 1.785x + 24.41 0.992 21.3 28 y = 2.760x + 2.063 0.979 21.9 29 y = 4.627x - 0.7116 0.966 13.6 30 y = 2.950x + 19.75 0.977 14.5 35 y = 0.6462x + 34.25 0.997 43.6 36 y = 0.3843x + 41.41 0.981 54.6 37 y = 2.787x + 16.55 0.981 16.4 38 y = 0.9919x + 37.73 1.000 24.9 39 y = 0.7431x + 38.04 0.999 32.8 40 y = 1.022x + 33.15 0.993 28.6 41 y = 1.137x + 33.32 0.990 28.2 56 y = 0.7326x + 34.94 1.000 37.5 57 y = 1.402x + 30.34 0.996 22.9 58 y = 5.331x - 6.109 0.976 12.9 Enox y = 5.331x + 25.10 0.988 7.0

[0297] The results of the PT experiment are shown in Table 3. It can be seen that the fucoidan sulfate compounds have no obvious effect on human plasma PT, indicating that they have no obvious inhibitory activity on the extrinsic coagulation pathway.

[0298] Table 3 Effects of fucoidan sulfate compounds on PT of human coagulation quality control plasma (mean±SD, n = 3)

[0299]

[0300] The results of the TT experiment are shown in Table 4. It can be seen that the fucoidan sulfate compounds have no obvious effect on human plasma TT, indicating that they have no obvious inhibitory activity on the common coagulation pathway.

[0301] Table 4 Effects of fucoidan sulfate compounds on TT of human coagulation quality control plasma (mean±SD, n = 3)

[0302]

[0303]

[0304] In summary, some fucoidan sulfate compounds can prolong human plasma APTT and have the activity of selectively inhibiting endogenous coagulation. Among them, fucoidan sulfates 29, 30, 37, and 58 (i.e., Formulas A - D of the present invention) have prominent anticoagulant activities, and the EC 2.0×They reached 13.6 μg / mL, 14.5 μg / mL, 16.4 μg / mL and 12.9 μg / mL respectively, which are comparable to the activities of existing clinical anticoagulants. In addition, all fucoidan sulfates had no obvious effect on PT and TT, indicating that the compounds mainly act on the intrinsic coagulation pathway and have no obvious effect on the extrinsic pathway and the common pathway.

[0305] The above are only the preferred embodiments of the present invention and do not impose any formal restrictions on the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A type of fucoidan sulfate compound, characterized in that: Having the structure shown in formula I or II: The fucoidan sulfate compound of the structure shown in Formula I includes the structure shown in any one of Formulas 1 to 30, E, F or G: The fucoidan sulfate compound of the structure shown in Formula II includes the structure shown in any one of Formulas 31 to 58:

2. The method for preparing the fucoidan sulfate compound according to claim 1, characterized in that: The following steps are involved: Using fucose as raw material, monosaccharide building blocks 1-4 were obtained; Replacing the 3-position protecting group of the monosaccharide building block 1-4 with an electron-donating group TBS to obtain a monosaccharide building block 1-6; Performing a deprotection reaction on the monosaccharide building block 1-6 to obtain a monosaccharide building block 1-8; Using the monosaccharide building blocks 1-4, 1-6 and 1-8, a one-pot synthesis reaction based on glycosyl donor pre-activation is performed to obtain Type I fucoidan fragments; polymerizing the Type I fucoidan oligosaccharide fragment to obtain a Type I fucoidan fragment, and removing the TBS temporary protecting group on the sugar chain of the Type I fucoidan fragment to obtain Type I fucoidan; Selectively removing the protecting group of the Type I fucoidan to obtain a Type I deprotected product; The 2-position on the sugar ring of the Type I deprotected product is subjected to sulfation modification to obtain the compounds 1 to 11; the 4-position on the sugar ring of the Type I deprotected product is subjected to sulfation modification to obtain the compounds 12 to 22; the 2-position and 4-position on the sugar ring of the Type I deprotected product are simultaneously subjected to sulfation modification to obtain the compounds 23 to 30 and compounds E, F and G; The monosaccharide building blocks 1-4 and 1-8 are respectively formed into PMB groups to obtain monosaccharide building blocks 1-10 and 1-12; Using the monosaccharide building blocks 1-6, 1-4, 1-10 and 1-12, a one-pot synthesis reaction based on glycosyl donor pre-activation is performed to obtain Type II fucoidan oligosaccharide fragments; Polymerizing the Type II fucoidan oligosaccharide fragment to obtain a Type II fucoidan fragment, and removing the TBS temporary protecting group on the sugar chain of the Type II fucoidan fragment to obtain Type II fucoidan; Selectively removing the protecting group of the Type II fucoidan to obtain a Type II deprotected product; The 2-position and 3-position on the sugar ring of the Type II deprotected product are simultaneously subjected to sulfation modification to obtain compounds 31 to 41; the 3-position and 4-position on the sugar ring of the Type II deprotected product are simultaneously subjected to sulfation modification to obtain compounds 42 to 52; the 2-position, 3-position and 4-position on the sugar ring of the Type II deprotected product are simultaneously subjected to sulfation modification to obtain compounds 53 to 58.

3. The preparation method according to claim 2, characterized in that: The temperatures for sulfation modification at the 2-position, sulfation modification at the 4-position, sulfation modification at both the 2-position and the 4-position, sulfation modification at both the 2-position and the 3-position, sulfation modification at both the 3-position and the 4-position, and sulfation modification at both the 2-position, the 3-position and the 4-position are independently room temperature, and the times are independently 7 to 14 hours.

4. The preparation method according to claim 2, characterized in that: The sulfation reagents for sulfation modification at the 2-position, sulfation modification at the 4-position, sulfation modification at both the 2-position and the 4-position, sulfation modification at both the 2-position and the 3-position, sulfation modification at both the 3-position and the 4-position, and sulfation modification at both the 2-position, the 3-position and the 4-position include pyridine sulfur trioxide and / or trimethylamine sulfur trioxide.

5. The preparation method according to claim 4, characterized in that: When the 2-position is sulfated, the 4-position is sulfated, the 2-position and 4-position are simultaneously sulfated, the 2-position and 3-position are simultaneously sulfated, the 3-position and 4-position are simultaneously sulfated, and the 2-position, 3-position and 4-position are simultaneously sulfated, the equivalent ratio of the sulfur trioxide pyridine to each naked hydroxyl group on the sugar ring in the raw material is 10:1, and the raw material is the Type I deprotection product or the Type II deprotection product.

6. The preparation method according to claim 2, characterized in that: After the sulfation modification at the 2-position, the sulfation modification at the 4-position, the sulfation modification at both the 2-position and the 4-position, the sulfation modification at both the 2-position and the 3-position, the sulfation modification at both the 3-position and the 4-position, and the sulfation modification at both the 2-position, the 3-position and the 4-position are completed, the obtained product is neutralized, and the alkaline substance used for neutralization includes sodium hydroxide.

7. Use of the fucoidan sulfate compound according to claim 1 in the preparation of anticoagulant active drugs.

8. The use according to claim 7, characterized in that: The dosage form of the anticoagulant active drug includes tablets, injections, capsules, granules, pills, powders, oral liquids, sustained-release preparations, controlled-release preparations or nano-preparations that are pharmaceutically acceptable.

9. The use according to claim 7, characterized in that: The anticoagulant active drug comprises an effective dose of the fucoidan sulfate compound, a pharmaceutically acceptable salt and a pharmaceutically acceptable carrier, auxiliary material, excipient and diluent.

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