RG-i type pectin lycium barbarum 63-saccharide and fragment thereof, preparation method therefor, and use thereof

Abstract

Provided in the present invention are a RG-I type pectin Lycium barbarum 63-saccharide and a fragment thereof, a preparation method therefor, and the use thereof. Specifically, the present invention provides a polysaccharide composition comprising a polysaccharide as shown in formula I or II or a salt thereof, a drug or health care product composition comprising the polysaccharide composition, and the use of the polysaccharide composition. In addition, the present invention also provides various intermediates for preparing the polysaccharide composition, a synthesis method for glycosidic bonds required for preparing the polysaccharide, and a method for preparing the polysaccharide composition.

Description

RG-I type pectin wolfberry 63 sugar and its fragments, their preparation methods and uses Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to RGI type pectin wolfberry 63 sugar and its fragments, as well as their preparation methods and uses. Background Technology

[0002] Chronic liver disease is a major global health concern, affecting over 800 million people. 70% of chronic liver disease patients also have liver fibrosis, approximately 20% of non-alcoholic steatohepatitis (NAHH) patients progress to fibrosis, and 15%-25% of liver fibrosis patients eventually develop irreversible cirrhosis. This trend has been increasing in recent years, making liver fibrosis a key focus and hot topic in contemporary medical research. However, there is a lack of safe and effective drugs for liver fibrosis globally, making the development of new drugs to effectively control the progression of liver fibrosis extremely necessary and important for the treatment of various liver diseases.

[0003] In conclusion, there is an urgent need in this field to develop safe and effective drugs for the treatment and prevention of liver fibrosis-related diseases. Summary of the Invention

[0004] The objective of this invention is to develop an efficient method for preparing RG-I type pectin wolfberry trisaccharide and its fragment oligosaccharides. This trisaccharide and the main chain oligosaccharide have anti-liver fibrosis activity and are expected to be developed into a candidate glycoside drug for the treatment of liver fibrosis.

[0005] In a first aspect of the invention, a polysaccharide composition is provided, wherein the polysaccharide composition contains a polysaccharide as shown in Formula I or a salt thereof;

[0006] in,

[0007] R 1 It is hydrogen, galactose, galactomannan or rhamnose galactomannan;

[0008] R 2 It is hydrogen, arabinose, or arabinogalactan;

[0009] R 3 It is hydrogen, galactose, galactan, or arabinogalactan;

[0010] R is hydrogen or -M 1 -(M 2 ) o -M 3 Where o is 0 or an integer between 1 and 30, M 1 The value is either "none" (does not exist) or C. 1-2 Alkylene, M 2Each is independently a divalent group selected from the following group: -OC 1-2 Alkylene-, C 1-2 Alkylene and M 3 Selected from the following groups: hydrogen, -NH2, -NH(C) 1-4 alkyl) and -NH(C 1-4 Alkyl)2.

[0011] In another preferred embodiment, the polysaccharide composition contains a polysaccharide of Formula I with a purity of ≥90%, preferably ≥95%, and more preferably ≥98%.

[0012] In another preferred embodiment, the polysaccharide composition contains ≥90% (preferably ≥95%; more preferably ≥98%) of a polysaccharide or a salt thereof as shown in Formula I, based on the total mass of the polysaccharide composition.

[0013] In another preferred embodiment, R 1 In this study, the degree of polymerization of galactomannan ranged from 1 to 11.

[0014] In another preferred embodiment, R 1 In this study, the degree of polymerization of rhamnose galactomannan ranged from 2 to 11.

[0015] In another preferred embodiment, R 1 Selected from the following group:

[0016] Where the subscript n11 is an integer from 1 to 11, the subscript n12 is an integer from 0 to 9, and the subscript n13 is an integer from 0 to 5.

[0017] In another preferred embodiment, R 2 In this study, the degree of polymerization of galactomannan ranged from 1 to 11.

[0018] In another preferred embodiment, R 2 In this study, the degree of polymerization of arabinogalactan ranged from 1 to 9.

[0019] In another preferred embodiment, R 2 for

[0020] Where the subscript n21 is an integer from 1 to 9, and the subscript n22 is an integer from 1 to 4.

[0021] In another preferred embodiment, R 3 In this study, the degree of polymerization of galactomannan ranged from 1 to 9.

[0022] In another preferred embodiment, R 3 In this study, the degree of polymerization of arabinogalactan ranged from 1 to 33.

[0023] In another preferred embodiment, R 3 for

[0024] Where n3 is an integer from 0 to 8.

[0025] In another preferred example, R is selected from the following group:

[0026] hydrogen, methyl,

[0027] Where o is an integer from 0 to 30.

[0028] In another preferred embodiment, o is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0029] In another preferred embodiment, R is

[0030] hydrogen or

[0031] In another preferred embodiment, the polysaccharide as shown in Formula I is polysaccharide 26 (main-chain decasaccharide compound) as shown below:

[0032] In another preferred embodiment, the polysaccharide as shown in Formula I is polysaccharide 1 as shown below (i.e., RG-I type pectin goji berry 63 sugar):

[0033] In a second aspect of the invention, a polysaccharide composition is provided, wherein the polysaccharide composition contains a polysaccharide as shown in Formula II or a salt thereof;

[0034] in,

[0035] R a for

[0036] hydrogen or

[0037] R b for

[0038] hydrogen or

[0039] Furthermore, R a and R b At least one of them is not hydrogen;

[0040] R represents hydrogen or -M1-(M2) o -M3: where o is 0 or an integer from 1 to 30, and M1 is none (does not exist) or C. 1-2 The alkylene groups M2 are each independently a divalent group selected from the group consisting of: -OC 1-2 Alkylene-, C 1-2 Alkyl groups and M3 are selected from the following group: hydrogen, -NH2, -NH(C 1-4alkyl) and -NH(C 1-4 Alkyl)2;

[0041] n can be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0042] In another preferred embodiment, R is defined as in Equation I.

[0043] In another preferred embodiment, n is 1, 2, 3, or 4.

[0044] In another preferred embodiment, R a for And R b It is hydrogen, or R a It is hydrogen and R b for R a for And R b for

[0045] In another preferred embodiment, R a For hydrogen or R b for

[0046] In another preferred embodiment, the polysaccharide composition contains a polysaccharide or a salt thereof as shown in Formula II-1;

[0047] In another preferred embodiment, the polysaccharide composition contains a polysaccharide as shown in Formula II-2 or a salt thereof;

[0048] In another preferred embodiment, the polysaccharide composition contains a polysaccharide of Formula II with a purity of ≥90%, preferably ≥95%.

[0049] In another preferred embodiment, the polysaccharide composition contains ≥90% (preferably ≥95%; more preferably ≥98%) of the polysaccharide or its salt as shown in Formula II, based on the total mass of the polysaccharide composition.

[0050] In another preferred embodiment, the polysaccharide as shown in Formula II is selected from polysaccharide 26 (main-chain decasaccharide compound), polysaccharide 36, and polysaccharide 37 as shown below:

[0051] In a third aspect of the invention, a pharmaceutical or health product composition is provided, wherein the pharmaceutical composition comprises:

[0052] (a) the polysaccharide composition as described in the first or second aspect; and

[0053] (b) A pharmaceutically or health product-acceptable carrier.

[0054] In a fourth aspect of the invention, the use of the polysaccharide composition as described in the first or second aspect is provided in the preparation of a medicament or health product for the treatment or prevention of liver fibrosis or complications arising from liver fibrosis.

[0055] In a fifth aspect of the invention, a polysaccharide intermediate is provided, wherein the intermediate is as shown in Formula III;

[0056] in,

[0057] R I a for

[0058] Hydrogen, R I 1 R I 8 or

[0059] R I b for

[0060] -OR I 1 R I 2 or

[0061] R I c For R I 1 Or R I 3 ;

[0062] Each R I 1 They are the same alcohol hydroxyl protecting groups;

[0063] R I 2 It is an -O-ol hydroxyl protecting group or leaving group;

[0064] Each R I 3 Each independently is different from R I 1 The alcohol hydroxyl protecting group;

[0065] Each R I 4 For the same carboxyl-hydroxy protecting group (such as benzyl (Bn), methyl (CH3), p-methoxybenzyl (PMB) or naphthylmethyl (Nap));

[0066] RI 5 Selected from the following groups: hydrogen, alcohol hydroxyl protecting group, -M I 1 -(M I 2 ) o -M I 3 Where o is 0 or an integer between 1 and 30, M 1 The value is either "none" (does not exist) or C. 1-2 Alkylene, M 2 Each is independently a divalent group selected from the following group: -OC 1-2 alkylene-, -C 1-2 alkylene- and M 3 It is hydrogen or -N(R) P )2; where R P C 1-4 Alkyl or amino protecting groups;

[0067] R I 8 It is hydrogen or different from R I 1 Protecting groups of alcohol hydroxyl groups (such as Lev);

[0068] R I 7 It is an alcohol hydroxyl protecting group.

[0069] In another preferred embodiment, R I 1 It is benzyl (Bn).

[0070] In another preferred embodiment, the -O-ol hydroxyl protecting group is selected from the group consisting of -O-allyl (All), -O-p-methoxyphenyl (MP), -O-p-methoxybenzyl (PMB), or -O-naphthylmethyl (Nap).

[0071] In another preferred embodiment, the leaving group is selected from the group consisting of: -SC 1-6 Alkyl groups (such as -SEt), -S-unsubstituted or with one or more C atoms 1-6 Alkyl-substituted phenyl groups (such as -SPh, -STol), -OTCAI, -OTBAI, and -OPTFAI.

[0072] In another preferred embodiment, the leaving group is selected from the group consisting of: -SC 1-6 Alkyl, -S-unsubstituted or with one or more C 1-6 Alkyl-substituted phenyl, -OTCAI, -OTBAI, or -OPTFAI.

[0073] In another preferred embodiment, each R I 3and R I 6 Each of them is independently selected from the following groups: Lev, Nap, and Fmoc.

[0074] In another preferred embodiment, the amino protecting groups are each independently Bn or Cbz.

[0075] In another preferred embodiment, R I 5 Selected from the following group:

[0076] methyl,

[0077] Where s is an integer from 0 to 30.

[0078] In another preferred embodiment, s is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0079] In another preferred embodiment, R I 5 for

[0080] hydrogen or

[0081] In another preferred embodiment, R I 5 -(CH2) s -N(Bn)Cbz; where s is an integer from 1 to 10 (preferably s is 2, 3, 4, 5, 6 or 7).

[0082] In another preferred embodiment, R I 7 For Bz.

[0083] In another preferred embodiment, depending on whether the branched sugars to be linked are the same, each R I 3 It can be the same as or different from R. I 1 R I 7 Alcohol hydroxyl protecting groups; that is, each corresponding R I 3 When the required glycosyl groups are consistent at the positions, then R I 3 It is the same as, but different from, R. I 1 R I 7 The alcohol hydroxyl protecting group; each corresponding R I 3 When the required glycosyl groups are different at different positions, then R I 3 It is different from RI 1 R I 7 The alcohol hydroxyl protecting group; each corresponding R I 3 When the sugar groups required to be attached at different positions are the same or different, then R I 3 It is also partially the same as, partially different from, R. I 1 R I 7 The alcohol hydroxyl protecting group.

[0084] In another preferred embodiment, the polysaccharide intermediate (as shown in Formula III) is shown in Formula III-1;

[0085] Among them, R I 1 R I 4 R I 5 R I 7 and R I c As defined in Equation III.

[0086] In another preferred example, in formula III-1, R I 1 It is Bn.

[0087] In another preferred embodiment, in formula III-1, R I 4 It is Bn.

[0088] In another preferred embodiment, in formula III-1, R I 5 For R I 5 -(CH2) s -N(Bn)Cbz; where s is an integer from 1 to 10 (preferably s is 2, 3, 4, 5, 6 or 7).

[0089] In another preferred embodiment, in formula III-1, R I 7 For Bz.

[0090] In another preferred embodiment, in equation III-1, each R I c Independently for R I 1 Or R I 3 And R I1 and R I 3 As defined in Equation III.

[0091] In another preferred embodiment, in equation III-1, each R I c The 0, 1, 2, 3, 4, or all of them (preferably 2 or 3, more preferably 3) in the range represent R. I 3 And the rest of R I c For R I 1 And R I 1 and R I 3 As defined in Equation III.

[0092] In another preferred embodiment, in formula III-1, R I 3 It is an alcohol hydroxyl protecting group, and R I 3 Unlike R I 1 R I 4 and R I 7 .

[0093] In another preferred embodiment, in equation III-1, each R I 3 They can be the same as or different from each other; preferably, each R... I 3 They are different from each other.

[0094] In another preferred embodiment, in formula III-1, R I 3 Each is an independent one of Lev, Nap, and Fmoc.

[0095] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III or III-1) is shown in Formula III-1a;

[0096] Among them, R I 1 R I 4 R I 5 and R I 7 As defined in Equation III-1; R I 31 R I32 and R I 33 The definition of R is the same as in equation III-1 I 3 Definition.

[0097] In another preferred embodiment, in formula III-1a, R I 31 R I 32 and R I 33 Each is an independent protecting group for a different alcohol hydroxyl group, and R I 31 R I 32 and R I 33 Both are different from R I 1 R I 4 and R I 7 .

[0098] In another preferred embodiment, in formula III-1a, R I 31 R I 32 and R I 33 Represent one of Lev, Nap, and Fmoc (preferably, R) I 31 For Lev, R I 32 For Nap and R I 33 For Fmoc).

[0099] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III, III-1 or III-1a) is compound 25;

[0100] In another preferred embodiment, the polysaccharide intermediate (as shown in Formula III) is shown in Formula III-2;

[0101] Among them, R I 1 R I 2 R I 4 and R I c As defined in Equation III; R I 8 The definition of R in Equation III is the same.I 3 Definition.

[0102] In another preferred embodiment, in formula III-2, R I c For R I 3 Or R I 1 And R I 1 and R I 3 As defined in Equation III; preferably, R I c For R I 3 .

[0103] In another preferred embodiment, in formula III-2, R I 3 It is an alcohol hydroxyl protecting group, and R I 3 Both are different from R I 1 R I 2 R I 4 and R I 8 .

[0104] In another preferred embodiment, in formula III-2, R I 1 It is Bn.

[0105] In another preferred embodiment, in formula III-2, R I 2 is -O-All, -O-PTFAI, or -O-TCAI.

[0106] In another preferred embodiment, in formula III-2, R I 4 It is Bn.

[0107] In another preferred embodiment, in formula III-2, R I 8 It is an alcohol hydroxyl protecting group, and R I 8 Unlike R I 1 R I 2 R I 4 and R I c .

[0108] In another preferred embodiment, in formula III-2, R I 8 It is one of Lev, Nap, and Fmoc; preferably, R I 8 For Lev.

[0109] In another preferred embodiment, in formula III-2, R I c It is one of Bn or Lev, Nap, and Fmoc; preferably, R I c One of Lev, Nap, and Fmoc; more preferably, R I c For Nap or Fmoc.

[0110] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III or III-2) is shown in Formula III-2a or III-2b;

[0111] Among them, R I 1 R I 2 R I 4 and R I 8 As defined in equation III-2; R I 32 and R I 33 As shown in equation III-2, R I 3 Definition.

[0112] In another preferred embodiment, in formula III-2a, R I 32 and R I 33 All are alcohol hydroxyl protecting groups, and R I 32 and R I 33 Both are different from R I 1 R I 2 R I 4 and R I 8 .

[0113] In another preferred embodiment, in formula III-2a, R I 32 and R I 33Are they the same or different (preferably, R) I 32 and R I 33 They are different.

[0114] In another preferred embodiment, in formula III-2a, R I 32 One of Lev, Nap, and Fmoc (preferably, R) I 32 For Nap).

[0115] In another preferred embodiment, in formula III-2a, R I 32 and R I 8 Represent one of Lev, Nap, and Fmoc (preferably, R) I 32 For Nap and R I 8 For Lev).

[0116] In another preferred embodiment, in formula III-2a, R I 33 One of Lev, Nap, and Fmoc (preferably, R) I 33 For Fmoc).

[0117] In another preferred embodiment, the polysaccharide intermediate (such as the polysaccharide intermediate shown in Formula III or III-2) is selected from the group consisting of: compound 7, compound 8, compound 16, and compound 17.

[0118] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III-2a) is compound 7 or compound 8.

[0119] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III-2b) is compound 16 or compound 17.

[0120] In another preferred embodiment, the polysaccharide intermediate (as shown in Formula III) is shown in Formula III-3;

[0121] Among them, R I 1 R I 2 R I 4 R I 7 and R I cAs defined in Equation III.

[0122] In another preferred embodiment, in equation III-3, each R I c 0, 1, or all (preferably 1) of the value is R. I 3 And the rest of R I c -for R I 1 And R I 1 and R I 3 As defined in Equation III.

[0123] In another preferred embodiment, R I 3 It is an alcohol hydroxyl protecting group, and R I 3 Unlike R I 1 R I 2 R I 4 and R I 7 .

[0124] In another preferred embodiment, in formula III-3, R I 1 It is Bn.

[0125] In another preferred embodiment, in formula III-3, R I 2 For -O-All, -O-PTFAI, or -O-TCAI; preferably, R I 2 For -O-All or -O-PTFAI.

[0126] In another preferred embodiment, in formula III-3, R I 4 It is Bn.

[0127] In another preferred embodiment, in formula III-3, R I 3 Each is an independent one of Lev, Nap, and Fmoc.

[0128] In another preferred embodiment, in formula III-3, R I c One of them is Lev, Nap, and Fmoc, and the other is Bn, or R. I c All are one of Lev, Nap, and Fmoc, or R Ic All are Bn.

[0129] In another preferred embodiment, in formula III-3, R I c One of them is Lev, Nap, and Fmoc, and the other is Bn.

[0130] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III or III-3) is shown in Formula III-3a;

[0131] Among them, R I 1 R I 2 R I 4 and R I 7 As defined in Equation III-3, R I 31 The definition of R is the same as in equation III-1 I 3 Definition.

[0132] In another preferred embodiment, in formula III-3a, R I 31 It is an alcohol hydroxyl protecting group, and R I 31 Both are different from R I 1 R I 2 R I 4 and R I 7 .

[0133] In another preferred embodiment, in formula III-3a, R I 31 One of Lev, Nap, and Fmoc (preferably, R) I 31 For Lev).

[0134] In another preferred embodiment, the polysaccharide intermediate (such as the polysaccharide intermediate shown in formula III, III-3 or III-3a) is compound 14 or 15;

[0135] In another preferred embodiment, the polysaccharide intermediate (as shown in Formula III) is shown in Formula III-4;

[0136] Among them, R I 1 RI 4 R I 5 and R I c As defined in Equation III; R I 8 The definition of R in Equation III is the same. I 3 Definition.

[0137] In another preferred embodiment, in formula III-4, R I c For R I 3 Or R I 1 (R is the preferred location) I 3 ); and R I 1 and R I 3 As defined in Equation III.

[0138] In another preferred embodiment, in formula III-4, R I 1 It is Bn.

[0139] In another preferred embodiment, in formula III-4, R I 4 It is Bn.

[0140] In another preferred embodiment, in formula III-4, R I 5 -(CH2) s -N(Bn)Cbz; where s is an integer from 1 to 10 (preferably s is 2, 3, 4, 5, 6 or 7).

[0141] In another preferred embodiment, in formula III-4, R I 8 It is an alcohol hydroxyl protecting group, and R I 8 Unlike R I 1 R I 2 R I 4 and R I c .

[0142] In another preferred embodiment, in formula III-4, R I 8 It is one of Lev, Nap, and Fmoc; preferably, R I 8 For Lev.

[0143] In another preferred embodiment, in formula III-4, R I 3 It is an alcohol hydroxyl protecting group, and R I 3 Both are different from R I 1 R I 2 R I 4 and R I 8 .

[0144] In another preferred embodiment, in formula III-4, R I 3 Each is an independent one of Lev, Nap, and Fmoc.

[0145] In another preferred embodiment, in formula III-4, R I c It is one of Bn or Lev, Nap, and Fmoc; preferably, R I c One of Lev, Nap, and Fmoc; more preferably, R I c For Nap or Fmoc.

[0146] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III or III-4) is shown in Formula III-4a;

[0147] Among them, R I 1 R I 4 R I 5 and R I 8 As defined in equation III-4; R I 33 As shown in equation III-4, R I 3 Definition.

[0148] In another preferred embodiment, in formula III-4a, R I 33 It is an alcohol hydroxyl protecting group, and R I 33 Unlike R I 1 R I 2 R I 4 and R I8 .

[0149] In another preferred embodiment, in formula III-4a, R I 33 One of Lev, Nap, and Fmoc (preferably, R) I 33 For Fmoc).

[0150] In another preferred embodiment, the polysaccharide intermediate (such as the polysaccharide intermediate shown in formula III, III-4 or III-4a) is compound 19;

[0151] In another preferred embodiment, the polysaccharide intermediate (as shown in Formula III) is shown in Formula III-5;

[0152] Among them, R I 1 R I 2 R I 4 and R I c As defined in Equation III; R I 8 The definition of R in Equation III is the same. I 3 Definition.

[0153] In another preferred embodiment, in equation III-5, each R I c Independently for R I 1 Or R I 3 And R I 1 and R I 3 As defined in Equation III.

[0154] In another preferred embodiment, in formula III-5, R I c 0, 1, or all (preferably 1) of the values ​​are R. I 3 And the rest of R I c For R I 1 And R I 1 and R I 3 As defined in Equation III.

[0155] In another preferred embodiment, in formula III-5, RI 1 It is Bn.

[0156] In another preferred embodiment, in formula III-5, R I 2 For -O-All, -O-PTFAI, or -O-TCAI; preferably, R I 2 For -O-All or -O-PTFAI.

[0157] In another preferred embodiment, in formula III-5, R I 4 It is Bn.

[0158] In another preferred embodiment, in formula III-5, R I 8 It is an alcohol hydroxyl protecting group, and R I 8 Unlike R I 1 R I 2 R I 4 and R I c .

[0159] In another preferred embodiment, in formula III-5, R I 8 It is one of Lev, Nap, and Fmoc; preferably, R I 8 For Lev.

[0160] In another preferred embodiment, in formula III-5, R I 3 It is an alcohol hydroxyl protecting group, and R I 3 Unlike R I 1 R I 2 R I 4 and R I 8 .

[0161] In another preferred embodiment, in formula III-5, R I 3 Each is independently one of Lev, Nap, and Fmoc; preferably, R I 3 Each can be either Nap or Fmoc.

[0162] In another preferred embodiment, in formula III-5, R Ic Each is independently one of Bn or Lev, Nap, and Fmoc; preferably, R I c One or all of them (preferably one) is one of Lev, Nap, and Fmoc (preferably Nap or Fmoc) and the rest are R I c It is Bn.

[0163] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III or III-5) is shown in Formula III-5a;

[0164] Among them, R I 1 R I 2 R I 4 and R I 8 As defined in Equation III-5; R I 32 The definition of R is the same as in equation III-5. I 3 Definition.

[0165] In another preferred embodiment, in formula III-5a, R I 32 It is an alcohol hydroxyl protecting group, and R I 3 Unlike R I 1 R I 2 R I 4 and R I 8 .

[0166] In another preferred embodiment, in formula III-5a, R I 32 It is one of Lev, Nap, and Fmoc; preferably, R I 32 For Nap.

[0167] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III, III-5 or III-5a) is compound 21 or 22;

[0168] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III) is shown in Formula III-6;

[0169] Among them, RI 1 R I 4 R I 5 and R I c As defined in Equation III; R I 8 The definition of R in Equation III is the same. I 3 Definition.

[0170] In another preferred embodiment, in equation III-6, each R I c Independently for R I 1 Or R I 3 And R I 1 and R I 3 As defined in Equation III.

[0171] In another preferred embodiment, in equation III-6, each R I c 0, 1, 2, or all of them (preferably 2 or 3, more preferably 2) represent R. I 3 And the rest of R I c For R I 1 And R I 1 and R I 3 As defined in Equation III.

[0172] In another preferred embodiment, in formula III-6, R I 1 It is Bn.

[0173] In another preferred embodiment, in formula III-6, R I 4 It is Bn.

[0174] In another preferred embodiment, in formula III-6, R I 5 -(CH2) s -N(Bn)Cbz; where s is an integer from 1 to 10 (preferably s is 2, 3, 4, 5, 6 or 7).

[0175] In another preferred embodiment, in formula III-6, R I 8 It is an alcohol hydroxyl protecting group, and R I8 Unlike R I 1 R I 4 and R I c .

[0176] In another preferred embodiment, in formula III-6, R I 8 It is one of Lev, Nap, and Fmoc; preferably, R I 8 For Lev.

[0177] In another preferred embodiment, in formula III-6, R I 3 It is an alcohol hydroxyl protecting group, and R I 3 Unlike R I 1 R I 4 and R I 8 .

[0178] In another preferred embodiment, in equation III-6, each R I 3 They can be the same as or different from each other; preferably, each R... I 3 They are different from each other.

[0179] In another preferred embodiment, in formula III-6, R I 3 Each can be independently one of Lev, Nap, and Fmoc; preferably Nap or Fmoc.

[0180] In another preferred embodiment, in equation III-6, each R I c 0, 1, 2, or all (preferably 2 or 3, more preferably 2) of R are one of Lev, Nap, and Fmoc (preferably Nap or Fmoc) and the rest are R I c Let Bn be the number of nodes; and R be the number of nodes. I 1 and R I 3 As defined in Equation III.

[0181] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III or III-6) is shown in Formula III-6a;

[0182] Among them, RI 1 R I 4 R I 5 and R I 8 As defined in Equation III-6; R I 32 and R I 33 In formula III-6, R I 3 Definition.

[0183] In another preferred embodiment, in formula III-6a, R I 32 and R I 33 It is an alcohol hydroxyl protecting group, and R I 32 and R I 33 Unlike R I 1 R I 2 R I 4 and R I 8 .

[0184] In another preferred embodiment, in formula III-6a, R I 32 and R I 33 Each of Lev, Nap, and Fmoc is a one of them; preferably, R I 32 For Nap and R I 33 For Fmoc.

[0185] In another preferred embodiment, the polysaccharide intermediate (such as the intermediate shown in Formula III, III-6 or III-6a) is compound 23;

[0186] In another preferred embodiment, at least one R I d For R I e .

[0187] In another preferred embodiment, when there are two or more R I d For R I e Each R I eThe hydroxyl protecting groups may be the same or different; preferably, each R I e These can be the same or different hydroxyl protecting groups.

[0188] In another preferred embodiment, R I a R I b R I c R I 1 R I 2 R I 3 R I 4 R I 5 R I 7 R I 8 M I 1 M I 2 M I 3 Subscript n and subscript o are each independently the corresponding groups in the intermediate compounds (e.g., compounds 25, 7, 8, 16, 17, 14, 15, 19, 21, 22, 23, etc.) prepared in the examples (e.g., Examples 1-6).

[0189] In another preferred embodiment, the polysaccharide backbone intermediate is used to prepare the polysaccharide composition as described in the first or second aspect.

[0190] In a sixth aspect of the invention, there is provided the use of a polysaccharide intermediate as described in the fifth aspect for preparing polysaccharide compositions as described in the first or second aspect, or for preparing polysaccharides containing α-GalA-(1→2)-Rha glycosidic bonds in disaccharide modules as shown in Formula IV.

[0191] In a seventh aspect of the invention, a method is provided for synthesizing α-GalA-(1→2)-Rha glycosidic bonds in a disaccharide module as shown in Formula IV.

[0192] Its characteristic is that it includes the following steps:

[0193] Among the various types,

[0194] R I 1 R I 2 RI 4 and R I c As defined in Equation III;

[0195] R I L It is a leaving group;

[0196] (S1) Glycosylate intermediates IV-1 and IV-2 to obtain intermediate IV-3;

[0197] (S2) The intermediate of formula IV-3 is subjected to a deprotection reaction to remove the >Si(tBu)2 protecting group, thereby obtaining the intermediate of formula IV-4;

[0198] (S3) Oxidize the IV-4 intermediate to obtain the IV-5 intermediate containing an α-GalA-(1→2)-Rha glycosidic bond.

[0199] In another preferred embodiment, R I L Selected from the following group: -SC 1-6 Alkyl groups (such as -SEt), -S-unsubstituted or with one or more C atoms 1-6 Alkyl-substituted phenyl groups (such as -SPh, -STol), -O-TCAI, -O-TBAI, and -O-PTFAI.

[0200] In another preferred embodiment, R I L -SC 1-6 Alkyl (e.g., -SEt) or -S- unsubstituted or with one or more C atoms 1-6 Alkyl-substituted phenyl groups (e.g., -SPh, -STol).

[0201] In another preferred embodiment, R I L -S- is not replaced or is occupied by one or more Cs 1-6 Alkyl-substituted phenyl groups (e.g., -SPh, -STol).

[0202] In another preferred embodiment, R I L -STol (i.e.) ).

[0203] In another preferred embodiment, R I 2 It is an -O-ol hydroxyl protecting group.

[0204] In another preferred embodiment, R I 2Selected from the group consisting of: -O-allyl (-O-All), -O-p-methoxyphenyl (-O-MP), -O-p-methoxybenzyl (-O-PMB), and -O-naphthylmethyl (-O-Nap).

[0205] In another preferred embodiment, R I 2 It is -O-allyl (-O-All).

[0206] In another preferred embodiment, R I 1 It is Bn.

[0207] In another preferred embodiment, R I c For R I 3 R I 3 As defined in Equation III.

[0208] In another preferred embodiment, R I c Selected from the following group: benzyl (Bn), naphthylmethyl (Nap), fluorenemethyloxycarbonyl (Fmoc), and 3-O-acetylpropionyl (Lev).

[0209] In another preferred embodiment, R I c Selected from the following group: naphthylmethyl (Nap), fluorenylmethoxycarbonyl (Fmoc), and 3-O-acetylpropionyl (Lev).

[0210] In another preferred embodiment, R I c It is naphthylmethyl (Nap).

[0211] In another preferred embodiment, R I 4 Selected from the following groups: benzyl (Bn), methyl (CH3), p-methoxybenzyl (PMB), naphthylmethyl (Nap).

[0212] In another preferred embodiment, the intermediate of formula IV-1 is shown below:

[0213] In another preferred embodiment, in step (S1), the glycosylation reaction is carried out in the presence of AgOTf.

[0214] In another preferred embodiment, in step (S1), the glycosylation reaction is carried out in the presence of Ar-SC1, wherein Ar is unsubstituted or converted by one or more (e.g., 1, 2, 3, or 4) R. S The C that was replaced 6-10 aryl or 5 to 10-membered heteroaryl; of which, R SEach of them independently selected from the following group: C 1-4 Alkyl, nitro.

[0215] In another preferred embodiment, Ar is phenyl.

[0216] In another preferred embodiment, Ar-SCl is selected from the group consisting of p-TolSCl, p-nitrophenylthiochloride, and phenylthiochloride.

[0217] In another preferred embodiment, in step (S1), the glycosylation reaction is carried out in the presence of a sterically hindered weak base.

[0218] In another preferred embodiment, the sterically hindered weak base comprises TTBP.

[0219] In another preferred embodiment, in step (S1), the glycosylation reaction is carried out in the presence of TTBP.

[0220] In another preferred embodiment, in step (S1), the glycosylation reaction is carried out in the presence of AgOTf and / or Ar-SCl and / or a sterically hindered weak base.

[0221] In another preferred embodiment, in step (S1), the glycosylation reaction is also carried out in the presence of a molecular sieve.

[0222] In another preferred embodiment, in step (S1), the glycosylation reaction is carried out in the presence of AgOTf, p-TolSCl, TTBP and molecular sieve.

[0223] In another preferred embodiment, in step (S2), the deprotection reaction is carried out in the presence of a fluoride.

[0224] In another preferred embodiment, the fluoride comprises: an HF / Py complex (hydrogen fluoride-pyridine complex), tetrabutylammonium fluoride, boron trifluoride-diethyl ether complex, or a combination thereof.

[0225] In another preferred embodiment, the fluoride is an HF / Py complex (hydrogen fluoride-pyridine complex).

[0226] In another preferred embodiment, in step (S2), the deprotection reaction is carried out in the presence of the HF / Py complex.

[0227] In another preferred embodiment, in step (S3), the oxidation reaction is carried out in the presence of an oxidant selected from the group consisting of TEMPO (2,2,6,6-tetramethylpiperidine-nitrogen oxide), BAIB (iodophenyldiacetic acid), or combinations thereof.

[0228] In another preferred embodiment, in step (S3), the oxidant is a combination of TEMPO and BAIB.

[0229] In another preferred embodiment, in step (S1), the glycosylation reaction is carried out in an inert solvent.

[0230] In another preferred embodiment, in step (S1), the inert solvent is DCM.

[0231] In another preferred embodiment, in step (S1), the equivalent ratio of intermediate of formula IV-1 to intermediate of formula IV-2 is 1 to 1.5:1; more preferably, it is 1.2:1.

[0232] In another preferred embodiment, in step (S1), the equivalence ratio of Ar-SCl (such as p-TolSCl) to the intermediate of formula IV-1 is 0.8 to 1.2:1; more preferably, 1:1.

[0233] In another preferred embodiment, in step (S1), the equivalence ratio of AgOTf to the intermediate of formula IV-2 is 2 to 4:1; more preferably, 3:1.

[0234] In another preferred embodiment, in step (S1), the equivalence ratio of the sterically hindered weak base (such as TTBP) to the intermediate of formula IV-2 is 0.8 to 1.2:1; more preferably, 1:1.

[0235] In another preferred embodiment, in step (S1), the reaction time of the glycosylation reaction is about 1 to 10 hours, preferably 3 to 5 hours.

[0236] In another preferred embodiment, in step (S2), the deprotection reaction is carried out in an inert solvent.

[0237] In another preferred embodiment, in step (S2), the deprotection reaction is tetrahydrofuran.

[0238] In another preferred embodiment, in step (S2), the equivalence ratio of the fluoride (such as the HF / Py complex) to the intermediate of formula IV-3 is 10 to 30:1; more preferably, 20:1.

[0239] In another preferred embodiment, in step (S2), the reaction time of the deprotection reaction is about 1 to 24 hours, more preferably 5 to 15 hours; and even more preferably about 10 hours.

[0240] In another preferred embodiment, in step (S3), the oxidation reaction is carried out in a mixed solvent of an inert solvent and water.

[0241] In another preferred embodiment, in step (S3), the inert solvent is DCM.

[0242] In another preferred embodiment, in step (S3), the equivalence ratio of TEMPO, BAIB and intermediate of formula IV-4 is 0.1 to 0.3:1 to 3:1; more preferably, it is 0.2:2:1.

[0243] In another preferred embodiment, in step (S3), the reaction time of the oxidation reaction is about 1 to 10 hours, more preferably 2 to 7 hours; and even more preferably about 5 hours.

[0244] In another preferred embodiment, step (S1) includes the following steps: adding the molecular sieve of formula IV-1 into an inert solvent and stirring (preferably, stirring for 10-60 min, such as 30 min), then adding AgOTf at -78°C and reacting (preferably, reacting for 1-20 min, more preferably, reacting for 2-7 min, best preferably, about 5 min), adding Ar-SCl and reacting (preferably, reacting for 1-20 min, more preferably, reacting for 2-7 min, best preferably, about 5 min) until the intermediate of formula IV-1 is completely consumed, adding the intermediate of formula IV-2 and a sterically hindered weak base to the obtained reaction system and reacting (1-10 hours, preferably 3-5 hours), gradually restoring to room temperature, and after post-treatment, obtaining the intermediate of formula IV-3.

[0245] In another preferred embodiment, in step (S1), the post-processing includes: quenching, filtration, washing, drying, rotary drying, and silica gel column chromatography.

[0246] In another preferred embodiment, step (S2) includes the steps of: dissolving the intermediate of formula IV-3 obtained in step (S1) in an inert solvent, adding fluoride, stirring at room temperature until the intermediate of formula IV-3 completely disappears (1 to 24 hours), and then post-processing to obtain intermediate of formula IV-4.

[0247] In another preferred embodiment, step (S2) includes: dilution with an inert solvent, washing the organic phase, evaporating under reduced pressure, and purification by silica gel column chromatography.

[0248] In another preferred embodiment, step (S3) includes the following steps: dissolving the intermediate of formula IV-4 obtained in step (S2) in a mixed solvent of an inert solvent and water, adding an oxidant, stirring at 0°C for 1 to 10 hours (preferably 2 to 7 hours; more preferably, about 5 hours), gradually restoring the reaction system to room temperature, and after post-treatment, obtaining intermediate of formula IV-5.

[0249] In another preferred embodiment, in step (S3), the post-processing includes: quenching, extraction, drying, vacuum drying, redissolving in a solvent (such as anhydrous DMF), adding benzyl bromide and potassium carbonate and stirring overnight, vacuum drying the reaction solution, and silica gel column chromatography purification.

[0250] In another preferred embodiment, the intermediate shown in Formula IV-5 can be formed into a polysaccharide backbone intermediate (such as the polysaccharide intermediate described in the fifth aspect of the present invention) by forming various glycosylation bonds with other monosaccharides or polysaccharides through glycosylation reaction methods known in the art or glycosylation reaction methods described in this application.

[0251] In another preferred embodiment, the intermediate shown in Formula IV-5 is used to prepare the polysaccharide intermediate shown in Formula III.

[0252] In another preferred embodiment, the intermediate shown in Formula IV-5 is used to prepare the polysaccharide composition as described in the first or second aspect.

[0253] In an eighth aspect of the invention, a method for preparing the polysaccharide composition as described in the first aspect is provided, comprising the steps of:

[0254] (1) Provide intermediate of formula III-1a

[0255] Among them, R I 1 R I 4 R I 5 and R I 7 As defined in Equation III-1; R I 31 R I 32 and R I 33 The definition of R is the same as in equation III-1 I 3 Definition;

[0256] (2) Make the intermediate R of formula III-1a 32 Positional deprotection; and optionally, the deprotected product is reacted with R S 1 -R S L The reaction yields branched polysaccharide intermediate 1.

[0257] (3) R of the branched polysaccharide intermediate 1 obtained in step (2) 31 Positional deprotection; and optionally, the deprotected product is reacted with R S 2 -R S L The reaction yields branched polysaccharide intermediate 2;

[0258] (4) R of the branched polysaccharide intermediate 2 obtained in step (3) 33Positional deprotection; and optionally, the deprotected product is reacted with R S 3 -R S L The reaction yields branched polysaccharide intermediate 3;

[0259] (5) Deprotection reaction is performed on branched polysaccharide intermediate 3 to obtain the polysaccharide composition as described in the first aspect;

[0260] Among them, R S L R is a leaving group; S 1 R S 2 and R S 3 R, where the hydroxyl group is protected. 1 R 2 and R 3 ;R 1 R 2 and R 3 As defined in Equation I.

[0261] In another preferred embodiment, the hydroxyl group being protected means that the hydroxyl group is protected by a protecting group selected from the group consisting of Bz, Bn, or a combination thereof.

[0262] In another preferred embodiment, R S L Selected from the following group: -SC 1-6 Alkyl, -S-unsubstituted or with one or more C 1-6 Alkyl-substituted phenyl; preferably, R S L -S- is not replaced or is occupied by one or more Cs 1-6 Alkyl-substituted phenyl groups.

[0263] In another preferred embodiment, R S L for

[0264] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0265] Figure 1. Compound 1 of trisaccharide, main chain decasaccharide 26 and polysaccharide LBP1C-2 inhibit the expression of fibrosis marker proteins induced by TGF-β in LX2 cells. Detailed Implementation

[0266] Through long-term and in-depth research, the inventors developed a sterically controlled glycosylation strategy with large steric hindrance protecting groups, combined with a post-oxidation strategy, to efficiently and selectively construct α-GalA-(1→2)-Rha glycosidic bonds. This led to the preparation of various disaccharide modules containing α-GalA-(1→2)-Rha glycosidic bonds. Furthermore, based on these disaccharide modules, other main-chain disaccharides, trisaccharides, tetrasaccharides, pentasaccharides, hexasaccharides, heptasaccharides, octasaccharides, and nonasaccharides were obtained by constructing other types of glycosidic bonds. As a result, for the first time, high-purity RG-I type pectin Lycium barbarum polysaccharide main-chain decasaccharide (i.e., polysaccharide 26) and other main-chain oligosaccharide fragments containing α-GalA-(1→2)-Rha glycosidic bonds (such as polysaccharides 36 and 37) were obtained. Furthermore, the disaccharide module containing α-GalA-(1→2)-Rha glycosidic bonds constructed in this application, and other polysaccharide modules prepared from it, can easily incorporate other protecting groups. This makes it highly suitable for introducing different protecting groups at specific positions, thereby enabling the introduction of side-chain oligosaccharides onto the main chain oligosaccharide of RG-I type pectin-goji berry polysaccharide, and ultimately obtaining high-purity RG-I type pectin-goji berry 63 sugar (i.e., polysaccharide 1) in high yield. In addition, the inventors have discovered that the main chain oligosaccharide of RG-I type pectin-goji berry polysaccharide exhibits superior activity compared to RG-I type pectin-goji berry 63 sugar. Based on these findings, the inventors have completed this invention.

[0267] the term

[0268] In this document, the abbreviations or terms have meanings well known to those skilled in the art. For example, some of the abbreviations used in this application can be found in the table below:

[0269] As used herein, when referring to a specific enumerated value, the term “about” means that the value can vary by no more than 1% from the enumerated values. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0270] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “composed of”.

[0271] Unless otherwise stated, the term "alkyl," either on its own or as part of another substituent, refers to a straight-chain or branched hydrocarbon group having a specified number of carbon atoms (i.e., C64 ... 1-6 (Indicates 1-4 carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, etc.

[0272] Unless otherwise stated, the term "alkylene", either on its own or as part of another substituent, refers to a divalent group derived from an alkane, such as -CH2-, -CH2CH2-, or -CH(CH3)-.

[0273] Unless otherwise stated, the term "aryl" refers to a polyunsaturated (usually aromatic) hydrocarbon group, which can be a monocyclic or fused together or covalently linked polycyclic ring. Generally, aryl groups have 6-10 ring atoms. The term "heteroaryl" refers to an aryl group (or ring) containing 1 to 5 heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. Generally, heteroaryl groups have 5-10 ring atoms, i.e., 5-10-membered heteroaryl groups, preferably 5-6 ring atoms, i.e., 5-6-membered heteroaryl groups, and contain 1, 2, 3, or 4 heteroatoms (such as N). Examples of aryl groups include phenyl, while examples of heteroaryl groups include pyridyl, etc.

[0274] As used in this article, chemical bonds marked with wavy lines or represented by dashed lines in the structural formula represent bonds that connect the group to other parts.

[0275] As used in this article, when a group or bracket subscript of 0 is in a chemical structural formula, it means that the group or the group within the bracket is absent (does not exist). For example, -(CH2)0-CH3 is equivalent to -CH3.

[0276] RG-I type pectin wolfberry polysaccharide

[0277] Pectin is a polysaccharide rich in galacturonic acid and has wide applications in the biomedical field. Lycium barbarum polysaccharide LBP1C-2 is an RG-I type pectin polysaccharide with the potential to develop anti-hepatic fibrosis drugs. The structure of polysaccharide LBP1C-2 consists of a highly branched repeating unit of an acidic 63-saccharide. Its main chain structure is composed of alternating α-galacturonic acid and α-rhamnose, with three different branched oligosaccharides attached to the 4-hydroxyl group of the rhamnose. Due to the complexity of this polysaccharide structure, there are currently no reports on its synthesis or related anti-hepatic fibrosis activity studies. Its chemical synthesis mainly faces the following challenges: 1) stereoselectively constructing α-GalA-(1→2)-Rha glycosidic bonds to assemble the main chain oligosaccharide; 2) assembling the polysaccharide using a rational glycosylation sequence; 3) stereoselectively and regioselectively introducing the three side-chain oligosaccharides.

[0278] This application develops a sterically hindered protecting group-controlled glycosylation method combined with a post-oxidation strategy to construct α-GalA-(1→2)-Rha glycosidic bonds in a highly efficient and specific manner, thereby preparing the main chain backbone. Furthermore, this application also develops a site-directed branched glycosylation method for the efficient synthesis of RG-I type pectin Lycium barbarum 63 sugar and related fragment compounds.

[0279] In one aspect, this application provides a polysaccharide composition containing a polysaccharide or a salt thereof as shown in Formula I (preferably, the polysaccharide composition contains ≥90% or ≥95% of a polysaccharide or a salt thereof as shown in Formula I, based on the total mass of the polysaccharide composition);

[0280] R, R1, R2, and R3 are defined as previously.

[0281] In some implementation schemes, R 1 Galactose, galactomannan, or rhamnose-galactomannan; and / or, R 2 It is arabinose or arabinogalactan; and / or, R 3 It is galactose, galactan, and arabinogalactan; and / or, R is a hydrogen atom or a linking arm such as an amino alkane chain or an amino PEG chain, or an alkyl group.

[0282] In one implementation, R 1 for

[0283] In one implementation, R 2 for

[0284] In one implementation, R 3 for

[0285] In one implementation, R is...

[0286] In another preferred embodiment, R, R 1 R 2 and R 3 Each of these is an independent corresponding group in the polysaccharide synthesized in the embodiments of this application.

[0287] In another aspect, this application provides a polysaccharide composition containing a polysaccharide as shown in Formula II or a salt thereof (preferably, the polysaccharide composition contains ≥90% or ≥95% of the polysaccharide as shown in Formula II or a salt thereof, based on the total mass of the polysaccharide composition);

[0288] Among them, R a R b And n is as defined above.

[0289] In another preferred embodiment, R a R b 'n' and 'n' are each independently the corresponding groups in the polysaccharides synthesized in the embodiments of this application.

[0290] In some embodiments, the polysaccharide shown in Formula II is as shown in Formula II-1.

[0291] The number of n is 0 to 9; R is a hydrogen atom or a linker arm such as an amino alkane chain or an amino PEG chain.

[0292] The anti-hepatic fibrosis activity test in this application showed that the 63 sugar (compound 1) and its main chain backbone oligosaccharide (compound 26) exhibited good activity and are expected to become candidate sugar drugs for the treatment of liver fibrosis-related diseases.

[0293] Synthesis method of Lycium barbarum polysaccharides

[0294] To address the significant challenges in synthesizing RG-I type pectin polysaccharides, this patent develops a convergent and stereocontrolled method to synthesize RG-I type pectin 63 sugar and its fragment derivatives modified with reduced terminal linker arms.

[0295] Among them, R 1 R 2 and R 3 As defined above.

[0296] Furthermore, addressing the challenge of constructing α-GalA-(1→2)-Rha glycosidic bonds in RG-I type pectin polysaccharides, this patent developed a sterically controlled glycosylation strategy with a large steric hindrance protecting group, combined with a post-oxidation strategy, to construct α-GalA-(1→2)-Rha glycosidic bonds in an efficient and specific manner, and prepared a multifunctional disaccharide module for the synthesis of main-chain oligosaccharides and their fragments.

[0297] In one aspect, a method is provided for synthesizing α-GalA-(1→2)-Rha glycosidic bonds in disaccharide modules as shown in Formula IV.

[0298] This includes the following steps:

[0299] Among the various types,

[0300] R I 1 R I 2 R I 4 and R I c As defined in Equation III;

[0301] R I L It is a leaving group;

[0302] (S1) Glycosylate intermediates IV-1 and IV-2 to obtain intermediate IV-3;

[0303] (S2) The intermediate of formula IV-3 is subjected to a deprotection reaction to remove the >Si(tBu)2 protecting group, thereby obtaining the intermediate of formula IV-4;

[0304] (S3) Oxidize the IV-4 intermediate to obtain the IV-5 intermediate containing an α-GalA-(1→2)-Rha glycosidic bond.

[0305] In some implementation schemes, R I 1 R I 2 R I 4 R I c and R I L And each step is as defined above.

[0306] In some implementations, the synthesis method is as follows:

[0307] In the above formula,

[0308] R 1 It can be a thioglycoside (such as SEt, SPh, STol, etc.) or a leaving group (such as -OTCAI, -OTBAI, or -OPTFAI, etc.).

[0309] R 2 It can be allyl (All), p-methoxyphenyl (MP), p-methoxybenzyl (PMB), or naphthylmethyl (Nap), etc.

[0310] R 3 It can be benzyl (Bn), naphthylmethyl (Nap), fluorenemethyloxycarbonyl (Fmoc), or 3-O-acetylpropionyl (Lev), etc.

[0311] R 4 It can be benzyl (Bn), methyl (CH3), p-methoxybenzyl (PMB), or naphthylmethyl (Nap), etc.

[0312] In other respects, this application also provides polysaccharide intermediates as shown in Formula III.

[0313] Among them, R I a R I b R I c R I 1 R I 2 R I3 R I 4 R I 5 R I 7 and R I 8 As defined above.

[0314] In other implementations, R I a R I b R I c R I 1 R I 2 R I 3 R I 4 R I 5 R I 7 and R I 8 Each of these refers independently to the specific functional group corresponding to each reaction intermediate or raw material in the examples.

[0315] RG-I type pectin wolfberry 63 sugar and main chain oligosaccharide fragments and their applications

[0316] As used herein, the terms “compound of the present invention,” “sugar of the present invention,” “polysaccharide of the present invention,” etc., refer to the polysaccharide represented by Formula I (including Lycium barbarum 63 sugar). The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates, or solvates of compounds of Formula (I).

[0317] The term "pharmaceutically acceptable salt" refers to a salt formed by the compounds of the present invention with an acid or base that is suitable for use as a medicine. Pharmaceutically acceptable salts include both inorganic and organic salts. A preferred class of salts are those formed by the compounds of the present invention with an acid. Acids suitable for forming salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid. Another preferred class of salts are salts formed by the compounds of the present invention with a base, such as alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., magnesium or calcium salts), ammonium salts (such as lower alkanol ammonium salts and other pharmaceutically acceptable amine salts), such as methylamine salts, ethylamine salts, propylamine salts, dimethylamine salts, trimethylamine salts, diethylamine salts, triethylamine salts, tert-butylamine salts, ethylenediamine salts, hydroxyethylamine salts, dihydroxyethylamine salts, trihydroxyethylamine salts, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0318] The RG-I type pectin wolfberry 63 sugar and main chain oligosaccharide fragment prepared in this patent exhibited good anti-liver fibrosis activity in a TGFβ-induced hepatic stellate cell-activated fibrosis model.

[0319] Since the polysaccharides of the present invention exhibit good anti-hepatic fibrosis activity in a TGFβ-induced hepatic stellate cell activation fibrosis model, the polysaccharides of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the polysaccharides of the present invention as the main active ingredient can be used to treat, prevent and alleviate liver fibrosis and other diseases caused by liver fibrosis.

[0320] The pharmaceutical compositions of the present invention comprise, within a safe and effective range, the compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, more preferably, 1-5000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0321] "Pharmaceutically acceptable carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0322] There are no particular limitations on the administration of the compounds or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and local administration.

[0323] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0324] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0325] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0326] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0327] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0328] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0329] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0330] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0331] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–5000 mg, preferably 20–500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skill of a skilled physician.

[0332] The main advantages of this invention include:

[0333] (1) The method of the present invention is very suitable for high-efficiency RG-I type pectin wolfberry 63 sugar and its fragment oligosaccharides.

[0334] (2) The high-purity RG-I type pectin wolfberry 63 sugar and main chain oligosaccharide prepared by the present invention have anti-liver fibrosis activity and are expected to be developed into a candidate sugar drug for the treatment of liver fibrosis.

[0335] (3) The method of the present invention is very suitable for synthesizing disaccharides and polysaccharide modules containing α-GalA-(1→2)-Rha glycosidic bonds.

[0336] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight.

[0337] Example 1

[0338] Step 1: First, take 1.2 equivalents of glucosinolate donor 2 and an equal mass of activated... Molecular sieves were dissolved in anhydrous DCM and stirred for 30 min under argon protection. Then, 3 equivalents of AgOTf were added at -78°C, and the reaction was allowed to proceed for 5 min. Next, 1.2 equivalents of p-TolSCl (p-toluenethiochloride) were added, and the reaction was allowed to proceed for another 5 min. TLC showed complete disappearance of the donor. One equivalent of acceptor 3 and one equivalent of TTBP (2,4,6-tri-tert-butylpyrimidine) were added to the reaction flask. The reaction was gradually allowed to return to room temperature over 3-5 hours. After TLC showed complete reaction, triethylamine was added to quench the reaction. The reaction solution was filtered through diatomaceous earth, washed with saturated sodium bicarbonate aqueous solution, and the organic layer was dried and then evaporated to dryness under reduced pressure. The resulting mixture was purified by silica gel column chromatography (eluent: petroleum ether and acetone), finally yielding compound 4 in 88% yield (purity >95%). 11H NMR (500 MHz, CDCl3) δ 7.85 - 7.74 (m, 4H), 7.49 - 7.42 (m, 7H), 7.38 - 7.27 (m, 11H), 5.90 (m, 1H, All - OCH2CHCH2), 5.31 - 5.24 (m, 1H, All - OCH2CHCH2), 5.21 - 5.17 (m, 1H, All - OCH2CHCH2), 5.03 (d, J = 11.1 Hz, 1H), 4.92 (d, J = 3.5 Hz, 1H, H1 - Gal), 4.89 - 4.76 (m, 4H, including H1 - Rha), 4.74 - 4.61 (m, 4H), 4.45 (d, J = 2.9 Hz, 1H, H4 - Gal), 4.20 - 4.15 (m, 1H, All - OCH2CHCH2), 4.12 (dd, J = 3.3, 1.8 Hz, 1H, H2 - Rha), 4.06 (dd, J = 10.0, 3.5 Hz, 1H, H2 - Gal), 4.00 - 3.89 (m, 4H, including All - OCH2CHCH2, H3 - Rha, H3 - Gal, H5 - Gal,), 3.85 (dd, J = 12.6, 1.7 Hz, 1H, H6 - Gal), 3.81 - 3.75 (m, 1H, H5 - Rha), 3.69 (dd, J = 12.6, 2.1 Hz, 1H, H6 - Gal), 3.61 (t, J = 9.4 Hz, 1H, H4 - Rha), 1.41 (d, J = 6.2 Hz, 3H, H6 - Rha), 1.03 (d, J = 7.1 Hz, 18H, tBu2Si). 13 13C NMR (125 MHz, CDCl3) δ 139.13, 138.6, 136.12, 134.09, 133.42, 133.09, 128.48, 128.39, 128.34, 128.21, 128.17, 128.01, 127.89, 127.8, 127.57, 127.54, 127.53, 127.11, 126.7, 126.16, 125.97, 117.35, 96.94 (C1 - Gal), 96.42 (C1 - Rha), 80.6 (C4 - Rha), 79.19 (C3 - Rha), 76.65, 75.45, 74.23 (C2 - Gal), 73.01 (C2 - Rha), 72.49, 72.03, 71.08 (C4 - Gal), 70.53, 68.4, 68.05, 67.38, 67.18 (C6 - Gal), 27.77, 27.46, 23.48, 20.78, 18.2 (C6 - Rha). HRMS (ESI) Calcd for C 55 H68 O 10 SiNH4 + [M+NH4] + 934.4920, measured value: 934.4919.

[0339] Step 2: Compound 4 was dissolved in an appropriate amount of tetrahydrofuran, and 20 equivalents of the HF / Py complex were added. The mixture was stirred at room temperature until the starting material was completely eliminated (approximately 10 hours). The reaction solution was diluted with an appropriate amount of DCM, and the organic phase was washed with a saturated sodium bicarbonate aqueous solution. The mixture after the organic phase was evaporated under reduced pressure was purified by silica gel column chromatography (eluent: petroleum ether and acetone). Compound 5 was finally obtained in 97% yield (purity greater than 95%). 1 H NMR(600MHz, CDCl3)δ7.85-7.76(m,4H),7.50-7.45(m,3H),7.43-7.27(m,15H),5.94-5 .85(m,1H,All-OCH2CHCH2),5.31-5.24(m,All-OCH2CHCH2),5.21-5.17(m,1H,All-OCH 2CHCH2),5.07(d,J=11.1Hz,1H),5.02(d,J=3.5Hz,1H,H1-Gal),4.87(d,J=1.9Hz,1H,H 1-Rha),4.85-4.64(m,7H),4.21-4.15(m,1H,All-OCH2CHCH2),4.09-4.05(m,2H,H5-Gal and H2-Rha),4.03(dd,J=3.3,1.4Hz,1H,H4-Gal),3.97-3.93(m,3H,All-OCH2CHCH2,H3-Gal,H3-Rha),3.90(dd,J=9.7,3.5Hz,1H,H2-Gal),3.82-3.78(m, 1H,H5-Rha),3.66(dd,J=11.8,5.4Hz,1H,H6-Gal),3.61(t,J=9.5Hz,1H,H4 -Rha), 3.58 (dd, J=12.1, 4.9Hz, 1H, H6-Gal), 1.40 (d, J=6.3Hz, 3H, H6-Rha). 13C NMR (150MHz, CDCl3) δ138.68,138.46,138.15,136.02,133.91,133.34,133.00,129.07,128.51,128.44,128.3 6,128.27,128.11,127.93,127.91,127.83,127.71,127.69,127.64,127.47,126.65,126.09,126.07,125.88, 117.29,96.83(C1-Gal),96.42(C1-Rha),80.41(C4-Rha),79.21(C3-Rha),76.92,75.74,75.29,74.27(C2-Rha ),72.64,72.57,72.18,69.27,69.15(C5-Gal),68.32,67.92,63.09(C6-Gal),18.17(C6-Rha).HRMS(ESI) calculated value: C 47 H 52 O 10 NH4 + [M+NH4] + 794.3899, Measured value: 794.3901.

[0340] Step 3: Compound 5 was dissolved in a DCM / water mixture (v / v = 2 / 1). 0.2 equivalents of TEMPO and 2 equivalents of BAIB were added under ice bath conditions. The mixture was stirred at 0°C for approximately 5 hours, and then the reaction system was gradually brought to room temperature. After TLC showed complete reaction, an appropriate amount of sodium thiosulfate was added to quench the reaction. After DCM extraction, the organic phase was dried over anhydrous sodium sulfate and evaporated under reduced pressure. The crude product was dissolved in anhydrous DMF, and 3 equivalents of benzyl bromide and 5 equivalents of potassium carbonate were added, followed by stirring overnight. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone), ultimately yielding compound 6 in 64% yield (purity > 95%). 11H NMR (500 MHz, CDCl3) δ 7.84 - 7.72 (m, 4H), 7.49 - 7.26 (m, 20H), 7.23 (dd, J = 5.0, 2.1 Hz, 3H), 5.91 - 5.83 (m, 1H, All - OCH2CHCH2), 5.27 - 5.22 (m, 1H, All - OCH2CHCH2), 5.20 - 5.15 (m, 2H, CH2 - COOBn - GalA and All - OCH2CHCH2), 5.06 - 5.01 (m, 2H, including H1 - GalA), 4.98 - 4.95 (m, 2H, H5 - GalA and CH2 - COOBn - GalA), 4.82 - 4.76 (m, 3H, including H1 - Rha), 4.74 - 4.61 (m, 4H), 4.56 (d, J = 12.3 Hz, 1H), 4.42 (dd, J = 3.2, 1.8 Hz, 1H, H4 - GalA), 4.17 - 4.13 (m, 1H, All - OCH2CHCH2), 4.10 (dd, J = 3.2, 1.9 Hz, 1H, H2 - Rha), 3.99 - 3.90 (m, 4H, All - OCH2CHCH2, H2 - GalA, H3 - GalA and H3 - Rha), 3.79 - 3.73 (m, 1H, H5 - Rha), 3.53 (t, J = 9.5 Hz, 1H, H4 - Rha), 1.38 (d, J = 6.2 Hz, 3H, H6 - Rha). 13 13C NMR (125 MHz, CDCl3) δ 168.81 (C6 - GalA), 138.58, 138.31, 138.03, 136.03, 135.51, 133.94, 133.39, 133.07, 128.60, 128.58, 128.47, 128.42, 128.39, 128.16, 128.01, 127.95, 127.80, 127.55, 127.53, 126.78, 126.19, 126.15, 125.96, 117.34, 97.32 (C1 - GalA), 96.53 (C1 - Rha), 79.93 (C4 - Rha), 79.01 (C3 - Rha), 76.57, 75.20, 74.96 (C2 - Rha), 74.68, 72.92, 72.63, 71.86, 70.26 (C5 - GalA), 68.65 (C4 - GalA), 68.34 (C5 - Rha), 67.94, 67.02, 18.20 (C6 - Rha). HRMS (ESI) Calcd: C 54 H 56 O 11 NH4+ [M+NH4] + 898.4161, Measured value: 898.4162.

[0341] Step 4: Dissolve compound 6 in anhydrous pyridine, add 20 equivalents of Lev2O and stir overnight. After TLC showed that the reaction was complete, evaporate the reaction solution under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone), and compound 7 was finally obtained in 90% yield (purity greater than 95%). 1 ¹H NMR (500MHz, CDCl₃) δ 7.84–7.74 (m, 4H), 7.50–7.41 (m, 3H), 7.40–7.26 (m, 20H), 5.94–5.84 (m, 2H, All-OCH₂CHCH₂ and H₄-GalA), 5.24–5.28 (m, 1H, All-OCH₂CHCH₂), 5.22–5.18 (m, 1H, All-OCH₂CHCH₂), 5.11–5.08 (m, 2H, including H₅-GalA), 5.06–5.03 (m, 2H, including H₁-GalA), 4.89–4.80 (m, 4H, including H₁-Rha). 4.73-4.67(m,2H),4.64-4.56(m,2H),4.45(d,J=11.1Hz,1H),4.19-4.15(m,1H,All-OCH 2CHCH2),4.11(dd,J=3.2,1.8Hz,1H,H2-Rha),4.03-3.92(m,3H,H3-GalA,All-OCH2CHCH 2, H3-Rha),3.82(dd,J=10.1,3.5Hz,1H,H2-GalA),3.80-3.74(m,1H,H5-Rha),3.51(t,J=9.4Hz,1H,H4-Rha), 2.65-2.43(m,4H,Lev-OCOCH2CH2COCH3),2.12(s,3H,Lev-OCOCH2CH2COCH3),1.39(d,J=6.3Hz,3H,H6-Rha). 13C NMR(125MHz, CDCl3)δ206.13(CO-Lev),171.46(CO-Lev),167.78(C6-GalA),138.68,138.23,138.03,135.90,135.11,133.88,133.32,133.02,129 .17,128.56,128.40,128.35,128.27,128.13,128.06,127.95,127.85,12 7.75,127.64,127.60,127.54,127.45,126.80,126.18,126.16,125.95,1 17.32,97.95(C1-GalA),96.56(C1-Rha),79.66(C4-Rha),79.17(C3-Rha) ,75.21(C2-Rha),75.15(C3-GalA),75.08(C2-GalA),74.42,72.84,71.94 ,71.85,69.44(C5-GalA),68.94(C4-GalA),68.32,67.84(C5-Rha),67.51,38.06(Lev),29.77(Lev),28.04(Lev),18.15(C6-Rha).HRMS(ESI) calculated value: C 59 H 62 O 13 NH4 + [M+NH4] + 996.4529, measured value: 996.4526.

[0342] Step 5: Compound 7 was dissolved in a THF / MeOH mixture (v / v = 1 / 1), and 0.5 equivalents of PdCl2 were added. The mixture was stirred at room temperature for 5 hours. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone). The resulting hemiacetal intermediate was dissolved in anhydrous DCM, and 2 equivalents of PTFAICl and 3 equivalents of cesium carbonate were added. The mixture was stirred at room temperature overnight. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to obtain compound 8, which was directly used in the subsequent glycosylation reaction.

[0343] Example 2

[0344] Step 1: First, add 1.2 equivalents of glucosinolate donor 2 and an equal mass of freshly activated... Molecular sieves were dissolved in anhydrous DCM and stirred for 30 min under argon protection. Then, 3 equivalents of AgOTf were added at -78°C, and the reaction was allowed to proceed for 5 min. Next, 1.2 equivalents of p-TolSCl were added, and the reaction was allowed to proceed for another 5 min. TLC showed complete disappearance of the donor. After adding 1 equivalent of acceptor 9 and 1 equivalent of TTBP to the reaction flask, the reaction was gradually allowed to return to room temperature. TLC showed complete reaction, and then triethylamine was added to quench the reaction. The reaction solution was filtered through diatomaceous earth, washed with saturated sodium bicarbonate aqueous solution, dried the organic layer, and then evaporated under reduced pressure. The resulting mixture was purified by silica gel column chromatography (eluent: petroleum ether and acetone), finally yielding compound 10 in 86% yield. 1 H NMR (600MHz, CDCl3) δ7.51-7.46 (dd, J=11.0, 7.5Hz, 4H), 7.40-7.28 (m, 16H), 5.96 -5.89(m,1H,OCH2CHCH2-All),5.33-5.28(m,1H,OCH2CHCH2-All),5.24-5.21(m,1 H,OCH2CHCH2-All),4.95-4.89(m,3H,including H1-Gal),4.87(d,J=1.7Hz,1H,H1-Rha),4 .85-4.75(m,2H),4.75-4.63(m,4H),4.49(d,J=2.9Hz,1H,H4-Gal),4.23-4.18(m, 1H),4.14(t,J=2.6Hz,1H,H2-Rha),4.10(dd,J=10.0,3.5Hz,1H,H2-Gal),3.99(td ,J=10.1,5.1Hz,3H),3.93(dd,J=9.4,3.2Hz,1H),3.87(dd,J=12.7,1.7Hz,1H,H6- Gal),3.81-3.76(m,1H,H5-Rha),3.71(dd,J=12.6,2.1Hz,1H,H6-Gal),3.59(t,J= 9.4Hz, 1H, H4-Rha), 1.42 (d, J=6.1Hz, 3H, H6-Rha), 1.06 (d, J=6.1Hz, 18H, tBu2Si). 13C NMR (150MHz, CDCl3) δ139.08,138.60,134.04,128.48,128.43,128.36,128.32,128. 17,128.04,127.86,127.78,127.52,127.04,117.31,96.82(C1-Gal),96.34(C1-Rha) ,80.57,79.16,76.60,75.41,74.22,72.89,72.51,72.03,71.10,70.57,68.33,68.01,67.33,67.11,27.74,27.44,23.45,22.78,20.75,18.07(C6-Rha).HRMS(ESI) calculated value: C 51 H 66 O 10 SiNH4 + [M+NH4] + 884.4764, Measured value: 884.4761.

[0345] Step 2: Compound 10 was dissolved in an appropriate amount of tetrahydrofuran solution, and 20 equivalents of HF / Py complex were added. The mixture was stirred at room temperature until the starting material was completely eliminated. The reaction solution was diluted with an appropriate amount of DCM, and the organic phase was washed with saturated sodium bicarbonate aqueous solution. The mixture after the organic phase was evaporated under reduced pressure was purified by silica gel column chromatography (eluent: petroleum ether and acetone). Compound 11 was finally obtained in 97% yield. 1 H NMR (600MHz, CDCl3) δ7.43-7.27(m,20H),5.91-5.85(m,1H,OCH2CHCH2-All),5.26-5.25(m,1H,OCH2CHCH2-All),5.20-5.17 (m,1H,OCH2CHCH2-All),5.01(d,J=3.5Hz,1H,H1-Gal),4.91(d,J=10.8Hz,1H),4.85(d,J=1.9Hz,1H,H1-Rha),4.81(d,J=11 .5Hz,1H),4.76(d,J=11.6Hz,1H),4.74-4.68(m,3H),4.65(dd,J=11.3,2.9Hz,2H),4.18-4.14(m,1H),4.08-4.00(m,3H),3. 97-3.87(m,4H),3.77-3.73(m,1H,H5-Rha),3.63(dd,J=11.7,5.2Hz,1H),3.59-3.52(m,2H),1.37(d,J=6.2Hz,3H,H6-Rha). 13C NMR (150MHz, CDCl3) δ138.76,138.61,138.57,138.25,134.00,128.59,128.51 ,128.44,128.09,128.00,127.93,127.80,127.75,127.72,127.52,117.36,96 .77(C1-Gal), 96.44(C1-Rha), 80.56, 79.24, 75.83, 75.37, 74.20, 72.71, 72.67, 72.27, 69.32, 69.26, 68.37, 67.99, 63.15, 18.16(C6-Rha). HRMS(ESI) calculated values: C 43 H 50 O 10 NH4 + [M+NH4] + 744.3742, Measured value: 744.3740.

[0346] Step 3: Compound 11 was dissolved in a DCM / water mixture (v / v = 2 / 1). Under ice bath conditions, 0.2 equivalents of TEMPO and 2 equivalents of BAIB were added, and the reaction system was gradually brought to room temperature. After TLC showed complete reaction, an appropriate amount of sodium thiosulfate was added to quench the reaction. After DCM extraction, the organic phase was dried over anhydrous sodium sulfate and evaporated under reduced pressure. The crude product was dissolved in anhydrous DMF, and 3 equivalents of benzyl bromide and 5 equivalents of potassium carbonate were added, followed by stirring overnight. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone), finally yielding compound 12 in 73% yield. 11H NMR (500 MHz, CDCl3) δ 7.43 - 7.26 (m, 18H), 7.25 - 7.21 (m, 2H), 5.92 - 5.84 (m, 1H, OCH2CHCH2-All), 5.27 - 5.23 (m, 1H, OCH2CHCH2-All), 5.22 - 5.14 (m, 2H, OCH2CHCH2-All and CH2-COOBn-GalA), 5.05 (d, J = 3.5 Hz, 1H, H1-GalA), 4.98 - 4.93 (m, 2H, CH2-COOBn-GalA, H5-GalA), 4.90 (d, J = 10.8 Hz, 1H, CH2-Bn), 4.83 - 4.72 (m, 4H, including H1-Rha and CH2-Bn), 4.64 (m, 3H, CH2-Bn), 4.55 (d, J = 12.2 Hz, 1H, CH2-Bn), 4.44 (dd, J = 3.4, 1.8 Hz, 1H, H4-GalA), 4.17 - 4.13 (m, 1H, All-OCH2CHCH2), 4.10 (dd, J = 3.3, 1.8 Hz, 1H, H2-Rha), 4.03 (dd, J = 9.8, 3.4 Hz, 1H, H3-GalA), 3.97 - 3.88 (m, 3H, H2-GalA, All-OCH2CHCH2, H3-Rha), 3.77 - 3.71 (m, 1H, H5-Rha), 3.49 (t, J = 9.5 Hz, 1H, H4-Rha), 1.37 (d, J = 6.3 Hz, 3H, H6-Rha). 13 13C NMR (125 MHz, CDCl3) δ 168.75 (C6-GalA), 138.52, 138.27, 138.01, 135.47, 133.89, 128.55, 128.45, 128.43, 128.36, 128.33, 128.30, 128.06, 128.02, 127.99, 127.95, 127.79, 127.74, 127.49, 127.43, 117.30, 97.16 (C1-GalA), 96.43 (C1-Rha), 80.18 (C4-Rha), 78.83 (C3-Rha), 76.50 (C3-GalA), 75.24 (C2-GalA), 74.99, 7,4.52 (C2-Rha), 72.90, 72.69, 71.84, 70.21 (C5-GalA), 68.68 (C4-GalA), 68.26 (C5-Rha), 67.89, 66.95 (CH2-COOBn-GalA), 18.06 (C6-Rha). HRMS (ESI) Calcd: C50 H 54 O 11 NH4 + [M+NH4] + 848.4004, measured value: 848.4009.

[0347] Step 4: First, add 1.5 equivalents of glucosinolate donor 13 and an equal mass of newly activated... Molecular sieves were dissolved in anhydrous DCM and stirred for 30 min under argon protection. Then, 5 equivalents of AgOTf were added at -50°C, and the reaction was allowed to proceed for 5 min. Next, 1.5 equivalents of p-TolSCl were added, and the reaction was allowed to proceed for another 5 min. TLC showed complete disappearance of the donor. After adding 1 equivalent of acceptor 12 and 1 equivalent of TTBP to the reaction flask, the reaction was gradually allowed to return to room temperature. TLC showed complete reaction, and then triethylamine was added to quench the reaction. The reaction mixture was filtered through diatomaceous earth, washed with saturated sodium bicarbonate aqueous solution, and evaporated under reduced pressure. The resulting mixture was purified by silica gel column chromatography (eluent: petroleum ether and acetone), finally yielding compound 14 in 69% yield. 11H NMR (600 MHz, CDCl3) δ 8.08 - 8.05 (m, 2H), 7.60 - 7.55 (m, 1H), 7.48 - 7.26 (m, 17H), 7.25 - 7.10 (m, 15H), 5.90 - 5.82 (m, 1H, All-OCH2CHCH2), 5.74 (t, J = 2.6 Hz, 1H, H2-Rha'), 5.28 - 5.21 (m, 2H, H1-Rha' and All-OCH2CHCH2), 5.19 - 5.16 (m, 1H, All-OCH2CHCH2), 5.10 (t, J = 9.8 Hz, 1H, H4-Rha'), 5.03 (d, J = 3.6 Hz, 1H, H1-GalA), 4.93 - 4.91 (m, 3H, H5-GalA and CH2-COOBn-B), 4.87 - 4.77 (m, 5H, including H1-Rha), 4.69 - 4.57 (m, 4H), 4.48 - 4.46 (m, 2H, including H4-GalA), 4.32 (d, J = 12.2 Hz, 1H), 4.15 - 4.12 (m, 1H, All-OCH2CHCH2), 4.04 - 4.02 (m, 2H, H3-GalA and H2-Rha), 3.97 - 3.91 (m, 2H, All-OCH2CHCH2 and H2-GalA), 3.89 (dd, J = 9.8, 3.2 Hz, 1H, H3-Rha'), 3.87 - 3.79 (m, 2H, H5-A and H3-Rha), 3.73 - 3.68 (m, 1H, H5-Rha), 3.43 (t, J = 9.5 Hz, 1H, H4-Rha), 2.74 - 2.69, (m, 1H, Lev), 2.62 - 2.49 (m, 2H, Lev), 2.41 - 2.37 (m, 1H, Lev), 2.14 (s, 3H, CH3-Lev), 1.34 (d, J = 6.2 Hz, 3H, H6-Rha), 1.21 (d, J = 6.2 Hz, 3H, CH3-Rha'). 13CNMR(150MHz, CDCl3)δ206.46(CO-Lev),172.07(CO-Lev),168.40(C6-GalA),165.44(C O-Bz-Rha'),138.53,138.48,138.29,138.22,138.04,134.83,133.87,133.21,130.07, 130.00,128.65,128.60,128.55,128.48,128.43,128.38,128.32,128.07,128.02,127. 92,127.84,127.80,127.77,127.71,127.58,127.51,127.37,117.36,99.63(C1-Rha'), 97.93(C1-GalA),96.62(C1-Rha),80.16(C4-Rha),78.81(C3-Rha),77.04,76.73,75.2 6,75.19,74.69,74.65,73.11(C4-Rha'),72.87,72.83,71.65,71.19,70.68,68.97(C2- Rha'),68.30(C5-Rha),67.84,67.46,67.42,37.94(Lev-OCOCH2CH2COCH3),29.91(CH3- Lev),28.07(Lev-OCOCH2CH2COCH3),18.09(C6-Rha),17.91(C6-Rha').HRMS(ESI) calculated value: C 75 H 80 O 18 NH4 + [M+NH4] + 1286.5683, measured value: 1286.5681.

[0348] Step 5: Compound 14 was dissolved in a mixed solution of THF / MeOH (v / v = 1 / 1), and 0.5 equivalents of PdCl2 were added. The mixture was stirred at room temperature for 5 hours. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone). The resulting hemiacetal intermediate was dissolved in anhydrous DCM, and 2 equivalents of PTFAICl and 3 equivalents of cesium carbonate were added. The mixture was stirred at room temperature overnight. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to obtain compound 15 (83%), which was directly used for subsequent glycosylation reactions.

[0349] Example 3

[0350] Step 1: Compound 7 was dissolved in a mixed solution of DCM / water (v / v = 20 / 1), and 1.1 equivalents of DDQ were added under ice bath conditions. The reaction system was gradually brought to room temperature. After 5 hours, TLC showed that the reaction was complete. The reaction solution was diluted with DCM, washed with saturated sodium bicarbonate, and the organic phase was evaporated to dryness. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to give an intermediate. The above intermediate was dissolved in a mixed solution of pyridine / DCM (v / v = 1 / 1), and 2 equivalents of FmocCl were added. The mixture was stirred overnight at room temperature. After TLC showed that the reaction was complete, the solvent was evaporated to dryness under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to give compound 16 in 79% yield. 1 H NMR (600MHz, CDCl3) δ7.80-7.76(m,2H),7.63-7.57(m,2H),7.41(t,J=7.5Hz,2H),7.37-7. 34(m,2H),7.33-7.26(m,9H),7.26-7.14(m,11H),5.93-5.83(m,2H,(m,2H,All-OCH2CHCH2 and H4-GalA),5.29-5.25(m,1H,All-OCH2CHCH2),5.23-5.20(m,All-OCH2CHCH2),5.07(d,J=1.7Hz,1H,H5-GalA),5.02-4.97(m,2H,including H1-GalA),4. 94(t,J=9.8Hz,1H,H4-Rha),4.86-4.80(m,2H,including H1-Rha),4.73(dd,J=11.3,9.7Hz,2H),4.61(dd,J=22.2,12.1Hz,2H),4.53(d,J=10.6Hz,1H),4. 49-4.38(m,3H),4.24(t,J=7.2Hz,1H),4.18-4.14(m,1H),4.13-4.08(m, 2H),3.98-3.94(m,1H),3.90(dd,J=9.9,3.3Hz,1H),3.86-3.82(m,1H,H5 -Rha),3.78(dd,J=10.1,3.6Hz,1H),2.58-2.34(m,4H,Lev-OCOCH2CH2COCH3),2.07(s,3H,Lev-OCOCH2CH2COCH3),1.28(d,J=6.2Hz,3H,H6-Rha). 13C NMR(150MHz, CDCl3)δ206.16(CO-Lev),171.36(CO-Lev),167.74(C6-GalA),154.95(CO-Fmoc),143.55,143.34,141.44,141.40,138.81,138 .11,137.82,135.16,133.70,129.01,128.50,128.42,128.38,128.36 ,128.30,128.21,128.01,127.99,127.86,127.64,127.61,127.58,127 .30,127.28,127.16,125.21,125.20,120.18,117.58,98.27(C1-GalA ),96.71(C1-Rha),77.44,76.19,75.40,74.96,74.39,72.95,72.19,71 .83,70.02,69.44,69.00,68.11,67.33(C5-Rha),66.76,46.95,38.10(Lev),29.80(CH3-Lev),28.04(Lev),17.49(C6-Rha).HRMS(ESI) calculated value: C 63 H 64 O 15 Na + [M+Na] + 1083.4137, measured value: 1083.4140.

[0351] Step 2: Compound 16 was dissolved in a THF / MeOH mixture (v / v = 1 / 1), and 0.5 equivalents of PdCl2 were added. The mixture was stirred at room temperature for 5 hours. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone). The resulting hemiacetal intermediate was dissolved in anhydrous DCM, and 2 equivalents of PTFAICl and 3 equivalents of cesium carbonate were added. The mixture was stirred at room temperature overnight. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to obtain compound 17 (73%), which was directly used for subsequent glycosylation reactions.

[0352] Step 3: Combine 1.5 equivalents of donor 17, 1 equivalent of acceptor 18, and an equal mass of newly activated... Molecular sieves were dissolved in anhydrous toluene and stirred for 30 min under argon protection. Then, 0.3 equivalents of TBSOTf were added at -20°C, and the reaction solution was gradually brought back to room temperature. After TLC showed complete reaction, an appropriate amount of pyridine was added to quench the reaction. The reaction solution was diluted with DCM, filtered, and the organic phase was washed with saturated sodium bicarbonate aqueous solution and concentrated. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to give compound 19 in 93% yield. 1 H NMR (600MHz, CDCl3) δ7.78 (dd, J=7.6, 2.4Hz, 2H), 7.60 (dd, J=18.3, 7.5Hz, 2H ),7.46-7.27(m,35H),7.25-7.12(m,14H),5.89(dd,J=3.5,1.7Hz,1H,H4-Gal A'),5.36(d,J=1.8Hz,1H,H1-Rha),5.34(d,J=12.2Hz,1H),5.20(d,J=13.5Hz ,2H),5.14-5.09(m,2H),5.03(d,J=12.0Hz,1H),4.99-4.89(m,3H, including H1-GalA and H1-GalA'),4.83(dd,J=19.6,11.9Hz,2H),4.74-4.61(m,6H),4.54-4.36(m,9H),4.30-4 .26(m,1H,H2-Rha),4.22(t,J=7.4Hz,1H),4.11(dd,J=10.1,3.4Hz,1H,H3-GalA'),3.99- 3.87 (m, 3H, including H3-Rha), 3.85-3.81 (m, 1H, H5-Rha), 3.68-3.54 (m, 2H), 3.47-3.37 (m, 1H), 3.31-3.15 (m, 2H), 2.59-2.39 (m, 4H), 2.10 (s, 3H), 1.66-1.46 (m, 4H), 1.36-1.20 (m, 5H). 13C NMR(150MHz, CDCl3)δ206.12(CO-Lev),171.31(CO-Lev),168.39(C6-GalA),167.68(C6-G alA),154.95,143.54,143.29,141.35,141.29,138.94,138.24,138.02,137.96,135.16,1 34.89, 128.95, 128.79, 128.71, 128.61, 128.57, 128.43, 128.34, 128.27, 128.22, 128.21, 128.11, 128.04, 128.00, 127.94, 127.91, 127.83, 127.78, 127.52, 127.37, 127.29, 127.2 5,127.21,126.86,125.24,120.07,98.83(C1-Rha),97.67(C1-GalA),97.49(C1-GalA),7 7.63,76.40,75.70,75.53,75.39,74.52,74.14,73.81,73.21,72.01,71.96,70.24,69.93 ,69.31,68.89,67.51,67.30,67.18,50.59,46.83,38.03(Lev-OCOCH2CH2COCH3),29.77(CH3-Lev),29.09,27.97(Lev-OCOCH2CH2COCH3),23.37,17.72(2×C6-Rha). HRMS(ESI) calculated values: C 107 H 109 O 23 NNa + [M+Na] + 1798.7283, Measured value: 1798.7286.

[0353] Step 4: Compound 19 was dissolved in a mixed solution of pyridine / acetic acid (v / v = 2 / 1), and 1.5 equivalents of hydrazine monohydrate were added. The mixture was stirred at room temperature. After 8 hours, TLC showed that the reaction was complete. The reaction solution was diluted with DCM, washed with saturated sodium bicarbonate aqueous solution, and the organic phase was concentrated by rotary evaporation. The solution was then purified by silica gel column chromatography (eluent: petroleum ether and acetone) to give compound 20 in 86% yield. 11H NMR (600 MHz, CDCl3) δ 7.74 (dd, J = 7.6, 2.4 Hz, 2H), 7.56 (dd, J = 17.4, 7.5 Hz, 2H), 7.40 - 7.26 (m, 22H), 7.25 - 7.06 (m, 27H), 5.31 (d, J = 1.8 Hz, 1H, H1-Rha), 5.29 (d, J = 12.3 Hz, 1H), 5.16 (d, J = 14.2 Hz, 2H), 5.08 (t, J = 12.4 Hz, 2H), 4.95 - 4.86 (m, 4H, including 2×H1-GalA), 4.81 - 4.73 (m, 3H), 4.67 - 4.56 (m, 5H), 4.50 - 4.30 (m, 10H), 4.24 (t, J = 2.5 Hz, 1H), 4.18 (t, J = 7.5 Hz, 1H), 4.00 (dd, J = 9.8, 3.3 Hz, 1H), 3.94 - 3.82 (m, 3H), 3.78 (dd, J = 9.9, 6.1 Hz, 1H, H5-Rha), 3.73 (dd, J = 9.8, 3.6 Hz, 1H), 3.61 - 3.49 (m, 1H), 3.42 - 3.32 (m, 1H), 3.25 - 3.13 (m, 2H), 1.61 - 1.44 (m, 4H), 1.30 - 1.20 (m, 5H, including H6-Rha). 13C NMR(150MHz,CDCl3)δ168.69(C6-GalA),168.44(C6-GalA),156.79(CO-Cbz),156.25(CO-Cbz ),155.00(CO-Fmoc),143.55,143.30,141.37,141.31,138.81,138.22,138.06,137.98,137. 95,136.94,135.50,134.91,128.81,128.73,128.64,128.59,128.52,128.49,128.46,128.26,128.20,128.17,128.10,128.07,128.04,127.98,127.93,127.88,127.72,127.44,127.38, 127.35,127.27,127.24,126.92,125.27,125.25,120.10,98.73(C1-Rha),97.66(C1-GalA), 96.87(C1-GalA),77.69,77.22,76.84,76.33,75.64,75.54,74.57,74.05,73.89,73.20,72. 67,71.99,71.86,70.26,70.09,69.95,68.87,68.44,67.53,67.30,67.26,66.71,50.59,50.30,47.17,46.84,46.18,29.77,29.12,28.01,27.59,23.39,17.75(C6-Rha). HRMS(ESI) calculated value: C 102 H 103 NO 21 NH4 + [M+NH4] + 1695.7361, Measured value: 1695.7358.

[0354] Example 4

[0355] Step 1: Combine 1.3 equivalents of donor 8, 1 equivalent of acceptor 12, and an equal mass of newly activated... Molecular sieves were dissolved in anhydrous DCM and stirred for 30 min under argon protection. Then, 0.3 equivalents of TBSOTf were added at -40°C, and the reaction solution was gradually brought back to room temperature. After TLC showed complete reaction, an appropriate amount of pyridine was added to quench the reaction. The reaction solution was diluted with DCM, filtered, and the organic phase was washed with saturated sodium bicarbonate aqueous solution and concentrated. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to give compound 21 in 58% yield.1 1H NMR (600 MHz, CDCl3) δ 7.84 - 7.70 (m, 4H), 7.48 - 7.44 (m, 4H), 7.41 - 7.27 (m, 24H), 7.25 - 7.13 (m, 20H), 5.91 - 5.82 (m, 2H, All - OCH2CHCH2 and H4 - GalA'), 5.39 (d, J = 1.8 Hz, 1H, H1 - Rha'), 5.25 - 5.21 (m, 1H, All - OCH2CHCH2), 5.19 - 5.17 (m, 1H, All - OCH2CHCH2), 5.13 - 5.08 (m, 2H, including H5 - GalA'), 5.07 (d, J = 3.5 Hz, 1H, H1 - GalA), 5.01 (d, J = 11.9 Hz, 2H), 4.98 (d, J = 3.7 Hz, 1H, H1 - GalA'), 4.96 - 4.92 (m, 2H, including H5 - GalA), 4.85 - 4.77 (m, 5H, including H1 - Rha), 4.74 (d, J = 11.9 Hz, 1H), 4.71 - 4.66 (m, 3H), 4.65 - 4.50 (m, 6H), 4.47 (d, J = 12.4 Hz, 1H), 4.38 (dd, J = 11.4, 2.2 Hz, 2H), 4.27 - 4.24 (m, 1H, H2 - GalA), 4.15 - 4.11 (m, 1H, All - OCH2CHCH2), 4.06 - 4.00 (m, 2H, including H2 - Rha), 3.98 - 3.88 (m, 4H), 3.85 (dd, J = 9.5, 3.1 Hz, 1H), 3.73 - 3.65 (m, 3H), 3.46 (t, J = 9.5 Hz, 1H, H4 - Rha'), 3.40 (t, J = 9.5 Hz, 1H, H4 - Rha), 2.64 - 2.45 (m, 4H), 2.13 (s, 3H), 1.34 (d, J = 6.2 Hz, 3H), 1.31 (d, J = 6.2 Hz, 3H). 13C NMR(150MHz,CDCl3)δ206.27(Lev),171.50(Lev),168.61(C6-GalA),167.86(C6-GalA'),138.87,138.53, 138.50,138.29,138.00,136.32,135.19,134.95,133.89,133.33,132.93,129.22,129.13,128.74,128.60 ,128.56,128.55,128.52,128.44,128.41,128.38,128.27,128.19,128.16,128.14,128.09,128.02,127.98,127.95,127.90,127.84,127.81,127.76,127.73,127.62,127.59,127.50,127.45,127.40,127.36,127. 33,127.26,126.40,126.15,126.02,125.86,117.33,98.31(C1-Rha'),97.54(C1-GalA),97.18(C1-GalA'),96.55(C1-Rha),80.22(C4-Rha),79.46(C4-Rha'),79.24,78.68,75.28,75.22,75.06,74.94,74.77,74.7 4,74.35,74.21,73.12,72.73,72.19,72.02,71.73,71.59,70.86,69.38,68.95,68.92,68.27,67.92,67.8 3,67.50,67.39,38.12(Lev),29.87(Lev),28.06(Lev),18.45(C6-Rha'),18.10(C6-Rha).HRMS(ESI) calculation value:C 106 H 110 O 23 NH4 + [M+NH4] + 1768.7776, actual measured value:1768.7771.

[0356] Step 2: Compound 21 was dissolved in a mixed solution of THF / MeOH (v / v = 1 / 1), and 0.5 equivalents of PdCl2 were added. The mixture was stirred at room temperature for 5 hours. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone). The resulting hemiacetal intermediate was dissolved in anhydrous DCM, and 2 equivalents of PTFAICl and 3 equivalents of cesium carbonate were added. The mixture was stirred at room temperature overnight. After TLC showed complete reaction, the reaction mixture was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to obtain compound 22 (68%), which was directly used for subsequent glycosylation reactions.

[0357] Step 3: Combine 1.4 equivalents of donor 22, 1 equivalent of acceptor 20, and an equal mass of newly activated... Molecular sieves were dissolved in anhydrous toluene and stirred for 30 min under argon protection. Then, 0.3 equivalents of TBSOTf were added at -40°C, and the reaction solution was gradually brought back to room temperature. After TLC showed complete reaction, a suitable amount of pyridine was added to quench the reaction. The reaction solution was diluted with DCM, filtered, and the organic phase was washed with saturated sodium bicarbonate aqueous solution and concentrated. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone), yielding compound 23 in 85% yield. 1 ¹H NMR (500MHz, CDCl₃) δ 7.84–7.76 (m, 4H), 7.72 (s, 2H), 7.60 (dd, J = 10.3, 7.6Hz, 2H), 7.48–7.26 (m, 49H), 7.25–7.04 (m, 48H), 5.88–5.83 (m, 1H), 5.41–5.29 (m, 4H, including 3×H₁-Rha), 5.19 (d, J = 9.3Hz, 2H), 5.12–4.77 (m, 21H, including 4×H₁-Gal). A),4.72-4.20(m,33H),4.04(dd,J=10.1,2.6Hz,1H),3.96-3.88(m,4H),3.87-3.56(m,10H),3.46-3.33(m,2H),3. 29-3.14(m,2H),2.64-2.44(m,4H,Lev),2.13(s,3H,CH3-Lev),1.63-1.45(m,4H),1.34-1.19(m,11H, including 3×H6-Rha). 13C NMR(125MHz,CDCl3)δ206.22(CO-Lev),171.47(CO-Lev),168.66(C6-GalA),168.49(C6-GalA),168.42(C6-GalA),167.85(C6-GalA),154.93(CO-Fmoc),143.60,143.35,141.39,141.33,138.89,138.85,138.81,138.59,138.52,138.48,138.29,138.09,138.01,137.99,137.92,136.33,135.19, 134.98,134.92,133.32,132.93,129.20,128.82,128.75,128.67,128.65,128.61,128.58,128.54,128.50,128.46,128.40,128.38,128.32,128. 28,128.25,128.22,128.16,128.14,128.11,128.02,127.95,127.89,127.82,127.79,127.73,127.69,127.60,127.56,127.53,127.49,127.44,12 7.38,127.34,127.30,127.26,127.20,126.89,126.42,126.12,126.05, 125.84,125.31,120.12,98.86(C1-Rha),98.19(C1-Rha),97.80(C1-Rha) ,97.69(C1-GalA),97.14(C1-GalA),97.06(C1-GalA),96.33(C1-GalA),80.11,79.52,79.36,78.93,77.76,76.29,75.77,75.68,75.02,74.93,74. 79,74.72,74.22,74.10,73.80,73.37,73.17,72.91,72.16,71.98,71.8 7,71.77,71.73,70.71,70.25,69.94,69.37,68.96,68.87,67.54,67.48, 67.29,67.09,53.54,50.92,50.31,47.18,46.85,38.11(Lev-OCOCH2CH2COCH3),29.84(CH3-Lev),29.12,28.07(Lev-OCOCH2CH2COCH3),23.40,18.43(C6-Rha), 18.26(C6-Rha), 17.81(C6-Rha). HRMS(ESI) calculated values: C. 205 H 207 NO 43 Na2 + [M+2Na] 2+ 1708.1913, Measured value: 1708.1913.

[0358] Step 4: Compound 23 was dissolved in a mixed solution of pyridine / acetic acid (v / v = 2 / 1), and 1.5 equivalents of hydrazine monohydrate were added. The mixture was stirred at room temperature. After 8 hours, TLC showed that the reaction was complete. The reaction solution was diluted with DCM, washed with saturated sodium bicarbonate aqueous solution, and the organic phase was concentrated by rotary evaporation. The solution was then purified by silica gel column chromatography (eluent: petroleum ether and acetone) to give compound 24 in 93% yield. 1 ¹H NMR (600MHz, CDCl₃) δ 7.87–7.75 (m, 6H), 7.64 (dd, J = 13.3, 7.5Hz, 2H), 7.50–7.28 (m, 61H), 7.26–7.11 (m, 36H), 5.45–5.33 (m, 4H, including 3×H₁-Rha), 5.24 (dd, J = 13.7, 5.6Hz, 3H), 5.10 (dd, J = 21.8, 11.8Hz, 2H), 5.05–4.92 (m, 11H, including 4×H₁-GalA), 4.89–4. 81(m,6H),4.76-4.63(m,11H),4.60-4.46(m,11H),4.46-4.31(m,9H),4.30-4.23(m,3H),4.09(dd,J=10.1,2.7Hz,1H),3.97 (m,4H),3.91-3.73(m,8H),3.69-3.52(m,3H),3.48-3.37(m,2H),3.32-3.20(m,2H),1.66-1.50(m,4H),1.37-1.26(m,11H). 13C NMR(150MHz,CDCl3)δ168.83(C6-GalA),168.66(C6-GalA),168.47(C6-GalA),168.40(C6-GalA),156.77(CO-Cbz),156.23(CO-Cbz),154.89(CO-Fm oc),143.55,143.31,141.34,141.29,138.85,138.77,138.65,138.52,13 8.48,138.24,138.06,137.97,137.93,137.89,137.86,136.92,136.80,13 6.36,135.48,134.94,134.92,134.87,133.29,132.89,128.80,128.72,128.64,128.62,128.60,128.58,128.55,128.46,128.43,128.41,128.38, 128.35,128.31,128.29,128.25,128.22,128.20,128.19,128.11,128.04,128.01,127.99,127.95,127.93,127.92,127.88,127.84,127.79,127.74 ,127.72,127.70,127.66,127.63,127.59,127.50,127.48,127.39,127.35,127.32,127.29,127.27,127.23,127.17,126.84,126.34,126.02,125.78,125.28,120.09,98.82(C1-Rha),98.05(C1-Rha),97.76(C1-Rha),97.66(C1-GalA),97.05(C1-GalA),96.51(C1-GalA),96.26(C1-GalA),80.06, 79.60,79.20,78.91,77.73,77.57,76.37,76.28,75.71,75.63,74.98,74.90,74.72,74.61,74.23,74.10,74.03,73.77,73.68,73.35,73.14,72.9 7,72.37,71.98,71.94,71.91,71.83,71.73,71.71,70.71,70.68,70.21,70.12,69.90,68.80,68.76,68.54,67.52,67.32,67.28,67.07,66.92,50.56, 50.27, 47.15, 46.80, 46.15, 29.09, 27.98, 27.56, 23.38, 18.41 (C6-Rha), 18.23 (C6-Rha), 17.79 (C6-Rha). HRMS (ESI) calculated value: C. 200 H 201 NO 41 Na2 + [M+2Na] 2+ 1659.1729, Measured value: 1659.1745.

[0359] Step 5: Combine 1 equivalent of donor 15, 0.7 equivalent of acceptor 24, and an equal mass of newly activated [substance / material]. Molecular sieves were dissolved in anhydrous toluene and stirred for 30 min under argon protection. Then, 0.2 equivalents of TBSOTf were added at -40°C, and the reaction solution was gradually brought back to room temperature. After TLC showed complete reaction, a suitable amount of pyridine was added to quench the reaction. The reaction solution was diluted with DCM, filtered, and the organic phase was washed with saturated sodium bicarbonate aqueous solution and concentrated. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone), yielding compound 25 in 90% yield. 11H NMR (600 MHz, CDCl3) δ 8.16 - 8.12 (m, 2H), 7.87 - 7.80 (m, 4H), 7.79 - 7.74 (m, 2H), 7.66 - 7.61 (m, 3H), 7.55 - 7.47 (m, 5H), 7.45 - 7.27 (m, 72H), 7.25 - 7.12 (m, 52H), 5.80 (t, J = 2.6 Hz, 1H), 5.49 (d, J = 1.8 Hz, 1H, H1 - Rha), 5.46 - 5.42 (m, 1H, H1 - Rha), 5.39 - 5.34 (m, 3H, including 2×H1 - Rha), 5.29 (d, J = 2.0 Hz, 1H), 5.26 - 5.17 (m, 3H), 5.12 (d, J = 12.2 Hz, 1H), 5.06 - 4.77 (m, 25H, including 5×H1 - GalA), 4.74 - 4.63 (m, 13H), 4.61 - 4.51 (m, 14H), 4.51 - 4.38 (m, 13H), 4.34 (d, J = 11.3 Hz, 1H), 4.31 - 4.21 (m, 5H), 4.09 - 4.03 (m, 2H), 4.00 - 3.72 (m, 16H), 3.69 - 3.38 (m, 5H), 3.33 - 3.21 (m, 2H), 2.83 - 2.46 (m, 4H, CH3COCH2CH2CO - Lev), 2.20 (s, 3H, CH3 - Lev), 1.66 - 1.50 (m, 4H), 1.41 - 1.20 (m, 17H, including 5×H6 - Rha). 13C NMR(150MHz,CDCl3)δ206.37(CO-Lev),172.06(CO-Lev),168.66(C6-GalA),168.62(C6-GalA),168.47(C6-GalA),168.40(C6-GalA),168.36(C6-Ga lA),165.46(CO-Bz),156.78(CO-Cbz),156.24(CO-Cbz),154.90(CO-Fmoc),143.58,143.32,141.36,141.31,138.87,138.80,138.72,138.69,138.6 1,138.54,138.50,138.27,138.17,138.07,137.99,137.94,137.88,136.82,136.37,134.97,134.95,134.90,134.85,133.31,133.19,132.90,130 .04,129.99,128.80,128.71,128.66,128.65,128.62,128.59,128.53,128.51,128.46,128.42,128.40,128.38,128.36,128.34,128.29,128.26,12 8.24,128.19,128.11,128.01,127.98,127.93,127.87,127.79,127.76,127.74,127.72,127.70,127.67,127.63,127.61,127.57,127.55,127.50, 127.46,127.44,127.42,127.38,127.36,127.33,127.27,127.23,127.16,126.86,126.32,126.02,126.01,125.78,125.29,120.09,99.61(C1-Rha) ,98.83(C1-Rha),98.04(C1-Rha),97.92(C1-Rha),97.77(C1-Rha),97.68(C1-GalA),97.03(C1-GalA),96.78(C1-GalA),96.59(C1-GalA),96.28(C1-GalA),80.09,79.97,79.70,79.15,79.10,78.92,77.73,77.59,76.76,76.27,75.75,75.66,75.10,75.06,74.90,74.79,74.62,74.44,74.40,74.35,74.24,74.00,73.79,73.74,73.60,73.35,73.14,72.85,72.82,72.78,71.93,71.89,71.84,71.82,71.74,70.99,70.77,70.72,70.68,70.50,70.23,69.91,68.93,68.81,67.51,67.44,67.37 ,67.25,67.07,50.59,50.30,47.17,46.83,46.17,37.91,29.89,29.10,28.06,27.58,23.38,18. 43(C6-Rha),18.37(C6-Rha),18.25(C6-Rha),17.87(C6-Rha),17.79(C6-Rha).HRMS(ESI) calculated value: C. 272 H 275 NO 58 Na2 + [M+2Na] 2+ 2264.4192, Measured value: 2264.4214.

[0360] Step 6: Dissolve compound 25 in a mixed solution of 1,4-dioxane / methanol / 1M sodium hydroxide (v / v / v = 3:1:1), maintain pH 10–11, and stir overnight at room temperature. After mass spectrometry analysis shows complete removal of all ester groups from the raw material, add Amberlite IR120 H + The reaction solution was neutralized with resin, filtered, and concentrated. The crude product was dissolved in a mixed solution of tetrahydrofuran / tert-butanol / water (v / v / v = 1:2:2), and Pd / C (10%) and Pd(OH)2 / C (50%), in equal masses to those of the starting material 25, were added. The mixture was stirred overnight under a hydrogen atmosphere. After mass spectrometry confirmed the completeness of the reaction, the mixture was filtered, and the filtrate was lyophilized. The crude product was then purified by C18 column chromatography (elution: water) to obtain the main-chain decasaccharide compound 26 (purity > 95%). 1H NMR(600MHz,D2O)δ5.27(m,5H,5×H1-Rha),5.01(s,4H,4×H1-GalA),4.96(s,1H ,H1-GalA),4.67(s,4H,H5-GalA),4.42(s,5H,H4-GalA),4.24(s,1H),4.12(s, 8H),4.06(s,2H),3.91(s,9H),3.80(s,6H),3.71(s,1H),3.58(s,1H),3.44-3. 33(m,5H),3.00(s,2H),1.68(s,4H),1.46(s,2H),1.30-1.21(m,15H,H6-Rha). 13 C NMR(150MHz,D2O)δ176.38(C6-GalA),176.17(C6-GalA),101.96(C1-Rha),99.86(4×C1-Rha),99.7 4(C1-GalA),98.85(4×C1-GalA),78.65,78.60,77.69,77.46,77.33,73.57,73.30,72.82,72.73,72 .22,71.99,71.81,71.79,71.63,71.45,70.76,70.42,70.15,69.41,69.31,69.26,40.80 (linker),29.49 (linker),27.77 (linker),23.81 (linker),18.09 (C6-Rha),18.00 (C6-Rha). HRMS (ESI) calculated values: C 65 H 103 NO 51 H + [M+H] + 1714.5570, Measured value: 1714.5573.

[0361] Example 5

[0362] Step 1: Compound 25 was dissolved in a DCM / water mixture (v / v = 20 / 1). 1.1 equivalents of DDQ were added under ice bath conditions. The reaction system was gradually brought to room temperature, and TLC showed complete reaction after 5 hours. The reaction solution was diluted with DCM, washed with saturated sodium bicarbonate, and the crude product obtained after rotary evaporation was purified by silica gel column chromatography (eluent: toluene and acetone) to give compound 27 in 77% yield. 11H NMR (500 MHz, CDCl3) δ 8.13 - 8.09 (m, 2H), 7.78 (dd, J = 7.6, 2.9 Hz, 2H), 7.63 - 7.59 (m, 3H), 7.49 (t, J = 7.7 Hz, 2H), 7.44 - 7.38 (m, 13H), 7.38 - 7.35 (m, 6H), 7.35 - 7.33 (m, 6H), 7.33 - 7.29 (m, 14H), 7.29 - 7.26 (m, 20H), 7.25 - 7.22 (m, 21H), 7.22 - 7.19 (m, 18H), 7.19 - 7.15 (m, 16H), 7.15 - 7.07 (m, 10H), 5.76 (t, J = 2.6 Hz), 5.47 - 5.44 (s, 1H, H1 - Rha), 5.42 (d, J = 1.7 Hz, 1H, H1 - Rha), 5.37 (s, 1H, H1 - Rha), 5.35 - 5.30 (m, 2H, including H1 - Rha), 5.26 (s, 1H, H1 - Rha), 5.22 - 5.15 (m, 3H), 5.09 (d, J = 12.2 Hz, 1H), 5.03 - 4.98 (m, 5H), 4.94 (dd, J = 9.2, 5.0 Hz, 6H), 4.92 - 4.88 (m, 3H), 4.88 - 4.85 (m, 3H), 4.84 - 4.80 (m, 3H), 4.79 - 4.77 (m, 2H), 4.70 - 4.66 (m, 7H), 4.63 (d, J = 10.4 Hz, 4H), 4.59 (d, J = 2.7 Hz, 3H), 4.57 - 4.54 (m, 3H), 4.53 - 4.45 (m, 10H), 4.43 - 4.29 (m, 11H), 4.27 - 4.19 (m, 5H), 4.08 - 4.02 (m, 2H), 3.99 - 3.90 (m, 5H), 3.87 (dt, J = 7.7, 4.7 Hz, 4H), 3.83 - 3.74 (m, 6H), 3.69 (dd, J = 9.3, 6.1 Hz, 1H), 3.66 - 3.52 (m, 6H), 3.48 - 3.34 (m, 3H), 3.30 - 3.16 (m, 2H), 2.81 - 2.41 (m, 4H), 2.16 (s, 3H), 1.62 - 1.48 (m, 4H), 1.32 - 1.20 (m, 17H). 13C NMR(125MHz,CDCl3)δ206.35(CO-Lev),172.04(CO-Lev),168.52(C6-GalA),168.50(C6-GalA),168.43(C6-GalA),168.37(C6-GalA),168.31(C6-Ga lA),165.43(CO-Bz),156.74(CO-Cbz),156.19(CO-Cbz),154.88(CO-Fmoc),143.53,143.29,141.33,141.28,138.80,138.76,138.64,138.58,138. 48,138.44,138.23,138.13,138.04,138.02,137.98,137.92,137.89,137.86,136.79,134.94,134.92,134.87,134.81,133.17,130.01,129.95,12 8.77,128.69,128.63,128.60,128.56,128.51,128.49,128.45,128.40,128.33,128.31,128.27,128.24,128.21,128.17,128.15,128.09,127.99,1 27.95,127.89,127.84,127.82,127.77,127.69,127.65,127.62,127.58,127.54,127.48,127.42,127.39,127.34,127.32,127.29,127.24,127.21,127.18,127.14,126.84,125.25,120.07,99.57(C1-Rha),98.80(C1-Rha),98.33(C1-Rha),98.06(C1-Rha),97.71(C1-Rha),97.64(C1-GalA),96. 97(C1-GalA),96.84(C1-GalA),96.30(C1-GalA),80.05,79.89,79.01,78.83,78.75,77.70,76.23,75.72,74.94,74.89,74.84,74.63,74.41,74.21,73.99,73.78,73.61,73.31,73.10,72.96,72.87,72.79,71.96,71.90,71.81,71.72,71.43,71.25,70.95,70.67,70.46,70.20,69.88,69.27,68.90,68.78,67.49,67.41,67.33,67.21,67.05,50.56,50.26,47.12,46.79,46.13,37.87,29.85,29.06,28.02,27 .54,23.34,18.27(C6-Rha),18.21(C6-Rha),18.06(C6-Rha),17.83(C6-Rha),17.76(C6-Rha).HRMS(ESI) calculated value: C. 261 H 267 O 58 NNa2 2+ [M+2Na] 2+ 2194.3879, measured value: 2194.3853.

[0363] Step 2: Combine 1.5 equivalents of donor 28, 1 equivalent of acceptor 27, and an equal mass of newly activated... Molecular sieves were dissolved in anhydrous toluene and stirred for 30 min under argon protection. Then, 1.3 equivalents of NIS and 0.2 equivalents of TBSOTf were added at -40°C, and the reaction solution was gradually brought back to room temperature. After TLC showed complete reaction, an appropriate amount of pyridine was added to quench the reaction. The reaction solution was diluted with DCM, filtered, and the organic phase was washed with saturated sodium bicarbonate aqueous solution and concentrated. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone, containing 20% ​​DCM), yielding compound 29 in 69% yield. 1H NMR(600MHz,CDCl3)δ8.17-8.14(m,2H),8.08-8.05(m,9H),8.04-8.01(m,12H),8.00-7.96(m,8H),7.84(dd,J=7.6,3.5Hz,2H),7.68-7.60(m,8H),7.58-7.51(m,7H),7.48-7.45(m,22H),7.44-7.41(m,22H),7.40-7.36(m,17H),7.36-7.33(m,15H),7.33-7.30(m,22H),7.29-7.27(m,19H),7.25-7.23(m,17H),7.23-7.21(m,16H),7.20-7.18(m,11H),7.17-7.16(m,7H),7.15-7.11(m,9H),6.97(t,J=7.6Hz,2H),6.89(t,J=7.3Hz,1H),5.83-5.79(m,2H),5.74(s,1H),5.65(dd,J=5.0,1.5Hz,1H),5.62-5.59(m,6H),5.56(d,J=2.2Hz,1H),5.53-5.51(m,3H),5.46-5.44(m,3H),5.43-5.41(m,3H),5.40(s,2H),5.37(d,J=12.5Hz,3H),5.28-5.23(m,6H),5.21(t,J=9.8Hz,1H),5.15(d,J=12.2Hz,1H),5.09-5.02(m,3H),4.99(dd,J=10.3,4.8Hz,6H),4.96-4.91(m,8H),4.89(dd,J=10.8,4.2Hz,4H),4.86-4.81(m,6H),4.77-4.71(m,8H),4.70-4.66(m,9H),4.64-4.60(m,8H),4.59-4.54(m,8H),4.52-4.49(m,9H),4.47-4.42(m,12H),4.40-4.37(m,2H),4.33-4.25(m,5H),4.21-4.15(m,3H),4.12-4.06(m,9H),4.04-3.97(m,6H),3.95-3.86(m,14H),3.83-3.77(m,3H),3.75-3.71(m,2H),3.69-3.62(m,5H),3.50-3.40(m,2H),3.35-3.22(m,2H),2.83-2.47(m,4H),2.21(s,3H),1.69-1.52(m,4H),1.45-1.23(m,17H). 13C NMR(150MHz,CDCl3)δ206.32(CO-Lev),172.04(CO-Lev),168.61(C6-GalA),168.51(C6-GalA),168.46(C6-GalA),168.38(C6-GalA),168.32(C6-GalA),166.18(CO-Bz),166.01(CO-Bz),165.99(CO-Bz),165.63(CO-Bz),165.59(CO-Bz),165.46(CO-Bz),165.42(CO-Bz),165.25(CO-Bz),165.18(CO -Bz),165.11(CO-Bz),165.08(CO-Bz),156.77(CO-Cbz),156.21(CO-Cbz),154.88(CO-Fmoc),143.56,143.31,141.34,141.29,138.90,138.87,138 .80,138.72,138.65,138.52,138.50,138.26,138.15,138.08,138.02,13 7.95,137.90,137.87,137.73,136.81,135.04,134.96,134.86,133.52,13 3.49,133.46,133.36,133.34,133.16,133.04,133.00,132.95,130.01,129.98,129.91,129.89,129.87,129.83,129.80,129.77,129.72,129.70, 129.63,129.61,129.56,129.54,129.44,129.25,129.20,129.08,128.98,128.95,128.78,128.70,128.65,128.63,128.61,128.57,128.55,128.52 ,128.49,128.47,128.44,128.36,128.34,128.32,128.28,128.25,128.23,128.19,128.17,128.14,128.09,128.03,128.00,127.98,127.95,127.9 0,127.85,127.77,127.74,127.72,127.69,127.67,127.65,127.58,127.55,127.48,127.41,127.38,127.34,127.28,127.24,127.21,127.14,126.85,126.74,125.27,120.07,107.12 (C1 - Ara),106.29 (C1 - Ara),106.21 (C1 - Ara),106.19 (C1 - Ara),106.14 (C1 - Ara),106.06 (C1 - Ara),105.49 (C1 - Ara),105.14 (C1 - Ara),99.57 (C1 - Rha),98.84 (C1 - Rha),97.82 (C1 - Rha),97.67 (C1 - Rha),97.04 (C1 - GalA),96.64 (C1 - GalA),96.54 (C1 - GalA),96.22 (C1 - GalA),83.51,83.42,83.10,82.95,82.76,82.67,82.55,82.10,82.03,82.00,81.76,81.71,81.64,81.56,81.49,80.54,80.41,80.09,79.94,79.25,79.02,78.83,77.78,77.74,77.71,76.70,76.26,75.80,75.72,75.10,74.90,74.76,74.69,74.41,74.30,74.22,73.98,73.89,73.78,73.60,73.34,73.13,72.84,72.79,72.11,72.00,71.96,71.91,71.82,71.80,71.76,71.71,71.56,70.95,70.71,70.62,70.41,70.22,69.88,68.91,68.77,68.18,67.50,67.39,67.31,67.22,67.06,66.02,65.61,65.52,65.35,65.26,63.70,63.56,53.52,50.57,50.28,47.15,46.81,46.16,37.88,29.86,29.74,29.08,28.05,27.56,23.36,18.38 (C6 - Rha),18.24 (C6 - Rha),18.21 (C6 - Rha),17.84 (C6 - Rha),17.78 (C6 - Rha). MS (MALDI - TOF) calculated value: C. 439 H 423 O 109 NNa + [M + Na] + 7474.7480, measured value: 7474.9510.

[0364] Step 3: Compound 29 was dissolved in a mixed solution of pyridine / acetic acid (v / v = 2 / 1), and 1.5 equivalents of hydrazine monohydrate were added. The mixture was stirred at room temperature. After 8 hours, TLC showed that the reaction was complete. The reaction solution was diluted with DCM, washed with saturated sodium bicarbonate aqueous solution, and the organic phase was concentrated by rotary evaporation. The solution was then purified by size exclusion chromatography (LH-20) (eluent: DCM and methanol) to give compound 30 in 93% yield. 1 H NMR (600MHz, CDCl3) δ8.16-8.12(m,2H),8.09-8.02(m,23H),7.99-7.86(m,10H),7.84(dd,J=7.6,3.4 Hz,2H),7.68-7.61(m,7H),7.58-7.52(m,6H),7.50-7.42(m,29H),7.41-7.36(m,25H),7.36-7.31(m, 31H),7.31-7.27(m,29H),7.25-7.20(m,37H),7.20-7.17(m,13H),7.17-7.10(m,11H),6.97(t,J=7.6 Hz,2H),6.91-6.86(m,1H),5.83-5.81(m,1H),5.80(s,1H),5.74(s,1H),5.65(dd,J=5.0,1.5Hz,1H), 5.60(dd,J=4.4,2.2Hz,6H),5.56(d,J=2.1Hz,1H),5.54-5.50(m,3H),5.47-5.44(m,2H),5.43-5.35( m,10H),5.29-5.24(m,6H),5.17-5.04(m,3H),5.02-4.93(m,12H),4.92-4.80(m,14H),4.76-4.68(m, 11H),4.66-4.60(m,19H),4.58-4.49(m,14H),4.47-4.36(m,14H),4.31-4.15(m,8H),4.13-4.04(m,9 H),4.02-3.89(m,10H),3.86-3.65(m,16H),3.49-3.22(m,4H),1.71-1.53(m,4H),1.44-1.24(m,17H). 13C NMR(150MHz,CDCl3)δ167.62(C6-GalA),167.52(C6-GalA),167.47(C6-GalA),167.41(C6-GalA),167.39(C6-GalA),165.20(CO-Bz),165.03(CO-Bz),165.01(CO-Bz),164.65(CO-Bz),164.60(CO-Bz),164.47(CO-Bz),164.43(CO-Bz),164.38(CO-Bz),164.27(CO-Bz),164.19(CO-Bz),164.09(CO -Bz),155.77(CO-Cbz),155.23(CO-Cbz),153.89(CO-Fmoc),142.58,142.33,140.36,140.30,137.92,137.75,137.63,137.53,137.28,137.10,13 6.99,136.91,136.88,136.82,136.75,134.06,133.95,133.90,132.53,1 32.51,132.47,132.38,132.36,132.30,132.16,132.05,132.02,131.96, 129.10,128.99,128.95,128.92,128.91,128.88,128.85,128.82,128.79,128.76,128.74,128.72,128.65,128.62,128.57,128.46,128.27,128. 22,128.10,127.99,127.96,127.80,127.71,127.67,127.65,127.62,127.59,127.56,127.53,127.50,127.48,127.45,127.42,127.40,127.38,12 7.36,127.33,127.30,127.27,127.25,127.19,127.16,127.04,127.01,126.99,126.92,126.91,126.86,126.83,126.79,126.75,126.71,126.69 ,126.66,126.60,126.58,126.56,126.51,126.49,126.41,126.37,126.34,126.30,126.25,126.23,126.13,125.86,125.75,124.29,119.09,106.13(C1-Ara),105.30(C1-Ara),105.23(C1-Ara),105.21(C1-Ara),105.16(C1-Ara),105.09(C1-Ara),104.51(C1-Ara),1 04.15(C1-Ara),98.31(C1-Rha),97.86(C1-Rha),96.91(C1-Rha),96.82(C1-Rha),96.68(C1-Rha),96.06(C1-GalA),95.7 8(C1-GalA),95.58(C1-GalA),95.24(C1-GalA),82.53,82.43,82.12,81.97,81.78,81.68,81.56,81.12,81.04,81.02,8 0.78,80.73,80.66,80.58,80.51,79.56,79.43,79.11,78.95,78.27,78.02,77.85,76.80,76.75,76.72,75.28,74.82,74 .74,74.30,74.10,73.91,73.77,73.44,73.40,73.23,72.94,72.79,72.60,72.35,72.15,72.11,71.80,71.71,71.12,71.02,70.97,70.93,70.81,70.73,70.56,70.30,69.72,69.65,69.54,69.24,68.90,68.00,67.79,67.42,67.19,66.51,66. 41,66.33,66.24,66.07,65.03,64.63,64.54,64.37,64.27,62.72,62.58,49.59,49.29,46.16,45.83,45.18,28.10,27.0 0,26.58,22.39,17.40(C6-Rha),17.26(C6-Rha),17.23(C6-Rha),17.17(C6-Rha),16.80(C6-Rha).MS(MALDI-TOF) calculated value: C. 434 H 417 O 107 NNa + [M+Na] + 7376.7112, measured value: 7376.8645.

[0365] Step 4: Combine 1.5 equivalents of donor 31, 1 equivalent of acceptor 30, and an equal mass of newly activated... Molecular sieves were dissolved in anhydrous toluene and stirred for 30 min under argon protection. Then, 10 equivalents of silver trifluoromethanesulfonate and 2.5 equivalents of iodine bromide were added at -40°C. The reaction solution was gradually restored to -30°C. After TLC showed complete reaction, a suitable amount of pyridine was added to quench the reaction. The reaction solution was diluted with DCM, filtered, and the organic phase was washed with saturated sodium bicarbonate aqueous solution and concentrated. The crude product was purified by silica gel column chromatography (eluent: toluene and acetone) to give compound 32 in 93% yield. 1 H NMR(500MHz, CDCl3)δ8.08(d,J=7.6Hz,2H),8.03-7.92(m,30H),7.87-7.71(m,4H),7.66-7.55(m ,8H),7.53-7.43(m,22H),7.41-7.28(m,122H),7.25-7.15(m,118H),7.14-7.04(m,55H),7.03-6 .95(m,19H),6.92-6.80(m,13H),5.74(s,1H),5.69(s,1H),5.60(dd,J=4.9,1.5Hz,1H),5.57-5. 51(m,10H),5.51-5.46(m,5H),5.41-5.35(m,9H),5.33-5.26(m,10H),5.24-5.19(m,6H),5.04-4. 95(m,9H),4.95-4.85(m,23H),4.83-4.73(m,9H),4.72-4.63(m,17H),4.61-4.52(m,31H),4.50- 4.41(m,21H),4.39-4.32(m,10H),4.32-4.22(m,23H),4.18-4.10(m,5H),4.08-4.02(m,7H),3.9 9-3.93(m,6H),3.91-3.82(m,15H),3.80-3.72(m,14H),3.70-3.61(m,15H),3.59-3.50(m,13H), 3.49-3.39(m,5H),3.37-3.26(m,16H),3.03-2.95(m,4H),1.62-1.48(m,4H),1.35-1.16(m,20H). 13C NMR(125MHz,CDCl3)δ168.65(C6-GalA),168.54(C6-GalA),168.49(C6-GalA),168.28(C6-GalA),168.05(C6-GalA),166.02(CO-Bz),165.61(CO-B z),165.38(CO-Bz),165.26(CO-Bz),165.13(CO-Bz),165.10(CO-Bz),165.03(CO-Bz),163.95(CO-Bz),163.85(CO-Bz),163.79(CO-Bz),163.66(CO -Bz),156.22(CO-Cbz),154.90(CO-Fmoc),138.95,138.74,138.71,138.59,138.52,138.49,138.09,138.06,137.90,137.80,137.73,137.49,135 .00,134.89,133.51,133.37,133.17,133.06,133.02,132.97,132.33,13 2.03,131.83,130.00,129.93,129.91,129.88,129.85,129.82,129.79,1 29.77,129.73,129.65,129.63,129.57,129.37,129.26,129.22,129.10,129.08,128.81,128.73,128.68,128.63,128.57,128.54,128.51,128.4 8,128.46,128.43,128.40,128.38,128.34,128.30,128.27,128.21,128.15,128.13,128.09,128.03,128.00,127.93,127.90,127.87,127.82,127 .79,127.76,127.71,127.69,127.64,127.60,127.57,127.49,127.46,127.28,127.13,126.87,126.74,126.67,121.09,120.10,119.82,107.86(C1-Ara),107.15(C1-Ara),106.31(C1-Ara),106.22(C1-Ara),106.16(C1-Ara),105.51(C1-Ara),105.15(C1-Ara),101.97,101.74(C1-Gal),101.28(C1-Gal),100.60(C1-β-Rha),98.55(C1-α-Rha),97.99(C1-α-Rha),97.82(C1-α-Rha),97.69 (C1-α-Rha),97.18(C1-GalA),97.12(C1-GalA),96.91(C1-GalA),95.62(C1-GalA),96.34(C1-G alA),83.43,83.11,82.94,82.78,82.56,82.11,82.03,81.74,81.66,81.58,81.51,80.55,80.4 3,79.90,79.72,79.28,78.93,78.58,78.36,78.19,78.08,77.80,77.75,75.81,75.40,75.16,74 .91,74.80,74.67,74.62,74.10,73.91,73.80,73.71,73.57,73.34,73.18,73.11,72.87,72.28,72.12,71.93,71.65,71.51,71.40,71.25,71.07,70.70,69.93,69.75,69.18,68.73,68.54,68 .00,67.41,67.24,67.06,66.03,65.63,65.37,65.26,63.72,63.58,47.16,29.10,28.00,27.59 ,23.38,18.41(C6-Rha),18.26(C6-Rha),18.18(C6-Rha),17.99(C6-Rha).MS(MALDI-TOF) calculated value: C. 731 H 703 O 172 NNa + [M+Na] + 12268.6186, measured value: 12268.9930.

[0366] Step 5: Compound 32 was dissolved in a dichloromethane solution containing 20% ​​triethylamine and stirred overnight at room temperature. After the reaction was confirmed to be complete by TLC, the reaction solution was concentrated, and the crude product was purified by size exclusion chromatography S-X1 (eluent: toluene and acetone) to give compound 33 in 85% yield. 1H NMR(800MHz,CDCl3)δ8.06(d,J=7.6Hz,2H),8.02-7.87(m,33H),7.86-7.82(m,2H),7.65-7.45(m,6H),7.44-7.27(m,97H),7.25-6.93(m,195H),6.93-6.77(m,40H),5.71(s,1H),5.66(s,1H),5.57(dd,J=5.0,1.5Hz,1H),5.55-5.43(m,12H),5.41-5.26(m,15H),5.26-5.14(m,9H),5.04-4.73(m,35H),4.71-4.16(m,98H),4.13-4.05(m,5H),4.04-3.40(m,81H),3.38-2.93(m,25H),1.63-1.45(m,4H),1.37-1.13(m,20H). 13C NMR(200MHz,CDCl3)δ168.67(C6-GalA),168.56(C6-GalA),168.51(C6-GalA),168.32(C6-GalA),168.08(C6-GalA),166.24(CO-Bz),166.05(CO-Bz),165.68(CO-Bz),165.64(CO-Bz),165.50(CO-Bz),165.46(CO-Bz),165.41(CO-Bz),165.29(CO-Bz),165.16(CO-Bz),165.13(CO-Bz),165.06(CO-Bz),163.98(CO-Bz),163.88(CO-Bz),163.82(CO-Bz),163.69(CO-Bz),139.17,138.98,138.80,138.74,138.63,138.56,138.53,138.50,138.36,138.13,138.10,138.08,137.99,137.93,137.86,137.83,137.79,137.77,137.53,135.05,135.00,134.92,133.56,133.54,133.50,133.40,133.20,133.09,133.05,132.99,132.36,132.06,131.85,130.03,129.96,129.94,129.92,129.88,129.86,129.83,129.80,129.77,129.76,129.69,129.66,129.60,129.41,129.37,129.30,129.26,129.14,129.10,129.00,128.76,128.72,128.66,128.62,128.60,128.57,128.54,128.51,128.49,128.47,128.43,128.41,128.38,128.36,128.33,128.30,128.27,128.24,128.23,128.18,128.17,128.12,128.07,128.06,128.03,127.99,127.96,127.93,127.91,127.89,127.85,127.82,127.80,127.78,127.76,127.73,127.72,127.67,127.63,127.59,127.52,127.49,127.40,127.37,127.31,127.14,126.92,126.77,126.70,107.19(C1-Ara),106.34(C1-Ara),106.26(C1-Ara),106.20(C1-Ara),106.17(C1-Ara),106.12(C1-Ara),105.54(C1-Ara),105.18(C1-Ara),102.05(C1-Gal),101.99(C1-Gal),101.94(C1-Gal),101.76(C1-Gal),101.31(C1-Gal),100.63(C1-β-Rha),98.59(C1-α-Rha ),98.06(C1-α-Rha),97.89(C1-α-Rha),97.71(2×C1-α-Rha),97.25(C1-GalA),97.20(C1-GalA),96.94(C1-GalA),96.71(C1-GalA),96.43(C1-GalA),83.45,83.13,82.96,82.81,82.72,82.59,82.14,82.07,82.05,81.79,81.76,81.68,81.61,81.54,80.59,80.46,79.93,79.76,79.32,79.23,78. 97,78.86,78.68,78.60,78.38,78.20,78.09,77.83,77.78,77.76,75.92,75.84,75.60,75.44,75.18,74.97,74.92,74.82,74.70,74.65,74.52,74 .33,74.27,74.18,74.12,74.03,73.94,73.83,73.74,73.60,73.57,73.43,73.36,73.22,73.14,72.93,72.90,72.85,72.78,72.61,72.31,72.15,7 2.02,71.97,71.92,71.72,71.67,71.63,71.55,71.43,71.28,71.22,71.11,70.93,70.79,70.73,70.60,70.33,69.97,69.78,69.36,69.22,68.74, 68.56,68.03,67.66,67.44,67.27,67.07,66.06,65.65,65.56,65.40,65.27,63.75,63.61,47.22,29.14,28.04,27.61,23.40,18.45(C6-Rha),18.35(C6-Rha),18.28(C6-Rha),18.21(C6-Rha),18.16(C6-Rha),18.02(C6-Rha).MS(MALDI-TOF) calculated value: C. 716 H 693 O 170 NNa + [M+Na] + 12046.5505, Measured value: 12046.8297.

[0367] Step 6: Combine 1.5 equivalents of donor 34, 1 equivalent of acceptor 33, and an equal mass of newly activated [substance / material]. Molecular sieves were dissolved in anhydrous toluene and stirred for 30 min under argon protection. Then, 10 equivalents of silver trifluoromethanesulfonate and 2.5 equivalents of iodine bromide were added at -40°C. The reaction solution was gradually restored to -30°C. After TLC showed complete reaction, a suitable amount of pyridine was added to quench the reaction. The reaction solution was diluted with DCM, filtered, and the organic phase was washed with saturated sodium bicarbonate aqueous solution and concentrated. The crude product was purified by silica gel column chromatography (eluent: petroleum ether and acetone) to give compound 35 in 33% yield. 1 H NMR(600MHz, CDCl3)δ8.08-7.86(m,110H),7.85-7.78(m,16H),7.63-7.5 2(m,27H),7.51-7.43(m,43H),7.42-7.32(m,91H),7.31-7.26(m,57H),7. 25-7.18(m,112H),7.17-7.05(m,222H),7.04-6.78(m,87H),5.72-5.41(m ,40H),5.40-5.26(m,51H),5.25-5.11(m,25H),5.08-4.91(m,12H),4.89- 4.79(m,16H),4.78-4.69(m,11H),4.69-4.52(m,61H),4.51-4.42(m,31H) ,4.41-4.30(m,39H),4.29-4.13(m,41H),4.11-4.01(m,37H),4.00-3.81( m,62H),3.79-3.69(m,29H),3.68-3.51(m,43H),3.49-3.38(m,21H),3.36 -3.20(m,22H),3.18-2.93(m,7H),1.61-1.44(m,4H),1.37-1.10(m,20H). 13C NMR(150MHz,CDCl3)δ168.09(C6-GalA),166.27(CO-Bz),166.07(CO-Bz),165.69(CO-Bz),165.63(CO-Bz),165.56(CO-Bz),165.50(CO-Bz),165.47 (CO-Bz),165.30(CO-Bz),165.26(CO-Bz),165.16(CO-Bz),165.13(CO-Bz),165.07(CO-Bz),164.87(CO-Bz),164.57(CO-Bz),163.98(CO-Bz),163.8 9(CO-Bz),163.81(CO-Bz),163.70(CO-Bz),139.47,139.31,138.94,138.76,138.69,138.61,138.55,138.52,138.45,138.43,138.18,138.11,138.09,138.07,138.05,137.98,137.91,137.85,137.83,137.81,137.78,137.75,137.52,135.39,135.05,134.94,134.84,133.61,133.56,133.52,13 3.31,133.18,133.10,133.06,133.01,132.87,132.39,131.92,130.04,129.96,129.92,129.88,129.86,129.83,129.80,129.78,129.73,129.67, 129.64,129.58,129.53,129.44,129.40,129.32,129.30,129.17,129.13,129.01,128.98,128.95,128.73,128.64,128.61,128.58,128.55,128.52 ,128.49,128.46,128.41,128.37,128.33,128.30,128.28,128.20,128.18,128.15,128.09,128.05,128.00,127.96,127.93,127.92,127.88,127.8 6,127.84,127.80,127.77,127.73,127.71,127.69,127.66,127.64,127.60,127.57,127.55,127.50,127.38,127.34,127.27,127.15,126.94,126.80,126.70,125.34,107.65 (C1 - Ara),107.14 (C1 - Ara),106.25 (C1 - Ara),106.20 (C1 - Ara),105.93 (C1 - Ara),105.52 (C1 - Ara),105.16 (C1 - Ara),102.00 (C1 - Gal),101.95 (C1 - Gal),101.86 (C1 - Gal),101.76 (C1 - Gal),101.66 (C1 - Gal),101.60 (C1 - Gal),101.31 (C1 - Gal),101.11 (C1 - Gal),101.05 (C1 - Gal),100.60 (C1 - β - Rha),98.45 (C1 - α - Rha),97.89 (C1 - α - Rha),97.42 (2×C1 - α - Rha),97.33 (C1 - α - Rha),96.57 (C1 - GalA),96.19 (C1 - GalA),95.98 (C1 - GalA),95.98 (C1 - GalA),95.90 (C1 - GalA),83.42,83.08,83.00,82.80,82.48,82.36,82.07,81.98,81.76,81.69,81.62,81.50,81.28,80.55,80.43,79.73,79.27,78.97,78.82,78.56,78.41,78.21,77.91,77.82,77.75,77.37,77.16,76.95,75.83,75.59,75.46,75.14,74.92,74.72,74.70,74.65,74.26,73.91,73.84,73.75,73.61,73.19,73.13,72.86,72.63,72.49,72.27,72.12,72.00,71.95,71.79,71.73,71.63,71.54,71.46,71.21,71.06,70.52,69.99,69.79,69.18,68.71,68.57,68.04,67.80,67.48,67.27,67.07,66.72,66.05,65.79,65.64,65.52,65.39,65.26,64.86,63.75,63.58,46.26,29.82,28.04,23.39,18.45 (C6 - Rha),18.31 (C6 - Rha),18.26 (C6 - Rha),18.18 (2×C6 - Rha),18.03 (C6 - Rha). MS (MALDI - TOF) calculated value: C.1422 H 1337 O 358 NNa + [M+Na] + 24174.6338, measured value: 24174.8914.

[0368] Step 7: Dissolve compound 35 in a mixed solution of dioxane / methanol / 1M sodium hydroxide aqueous solution (v / v / v = 5:2:2), maintaining the pH at 10–11. After stirring overnight at room temperature, add Amberlite IR120 H + The resin-neutralized reaction solution was filtered, concentrated, and the crude product was filtered through size exclusion chromatography (LH-20) (eluent: methanol and DCM) to generate an intermediate. At -78°C, approximately 300 mg of metallic sodium was added to 10 mL of liquid ammonia. After 5 minutes, a tetrahydrofuran solution of the above intermediate was slowly added dropwise, and the mixture was slowly brought back to room temperature. After approximately 4 hours, 0.5 mL of isoprene and 5 g of ammonium chloride solid were added to quench the reaction. The resulting crude product was purified by graphite column chromatography to remove salt and generate an intermediate (eluent: acetonitrile / 10 mmol ammonium bicarbonate aqueous solution, v / v = 1:1). The obtained intermediate was redissolved in a mixed solution of dioxane / methanol / 1 M sodium hydroxide aqueous solution (v / v / v = 1:1:1), maintaining the pH at 10–11. After stirring overnight at room temperature, Amberlite IR120 H was added. + The reaction solution was neutralized with resin, filtered, and concentrated. The crude product was dissolved in pure water, and an appropriate amount of Pd / C (10%) was added. The mixture was stirred overnight under a hydrogen atmosphere. After the reaction was confirmed to be complete by mass spectrometry, the mixture was filtered, and the filtrate was lyophilized. The crude product was then purified by a Bio-Gel P10 column (elution: 0.1M NH4HCO3) to obtain 63 sugar compound 1 (purity greater than 95%). 1H NMR(800MHz,D2O)δ5.24(s,2H,2×H1-Ara),5.18(s,6H,4×H1-Ara and 2×H1-Rha),5.16(s,3H,3×H1-Rha),5.14(s,1H,H1-Ara),5.09-5.07(m,3H,3×H1-Ara),5.05(s,1H,H1-Ara),5.03(s,1H,H1-Ara),5.02-5.00(m,21H,21×H1-Ara),4.97-4.92(m,5H,4×H1-GalA and H1-Rha-β),4.89(s,1H,H1-GalA),4.63-4.59(m,12H,7×H1-Gal and 5×H5-GalA),4.54(d,J=7.7Hz,2H,2×H1-Gal),4.48-4.44(m,5H,5×H4-GalA),4.42-4.33(m,10H,10×H1-Gal),4.26(td,J=5.3,3.0Hz,2H),4.24-4.20(m,5H),4.18-4.11(m,36H),4.09-4.00(m,48H),3.99-3.91(m,37H),3.90-3.62(m,157H),3.61-3.54(m,13H),3.54-3.42(m,10H),3.36-3.26(m,7H),2.92(t,J=7.8Hz,2H,linker),1.56-1.68(4H,linker),1.40-1.34(m,2H,linker),1.26-1.16(m,18H,6×CH3-Rha). 13C NMR(200MHz,D2O)δ174.79(C6-GalA),109.99(C1-Ara),108.11(C1-Ara),108.01(C1-Ara), 107.78(C1-Ara),107.67(C1-Ara),104.94(C1-Gal),104.56(C1-Gal),104.07(C1-Gal),104 .03(C1-Gal),103.82(C1-Gal),103.77(C1-Gal),103.72(C1-Gal),100.26(C1-β-Rha),99.1 7(C1-α-Rha),98.94(C1-GalA),98.22(C1-GalA),84.55,84.49,82.94,82.54,82.39,82.11, 81.85, 81.80, 81.53, 81.50, 81.43, 81.40, 81.38, 79.82, 79.62, 77.32, 77.12, 75.73, 75.66, 75.28, 74.35, 73.92, 73.34, 73.20, 72.79, 72.63, 71.62, 71.35, 71.15, 70.84, 70.46, 69.82 ,69.22,69.00,68.46,67.47,67.35,61.77,61.73,61.69,61.62,61.56,61.49,39.97,28.62,26.92,22.96,17.60(C6-Rha),17.28(C6-Rha),17.17(C6-Rha).HRMS(ESI,negative) calculated values: C 350 H 563 O 282 N 4- [M-4H] 4- 2322.9931, measured value: 2322.9893.

[0369] Example 6

[0370] Step 1: Dissolve compound 20 in a mixed solution of 1,4-dioxane / methanol / 1M sodium hydroxide (v / v / v = 3:1:1), maintain pH 10–11, and stir overnight at room temperature. After mass spectrometry analysis shows complete removal of all ester groups from the raw material, add Amberlite IR120 H... +The reaction solution was neutralized with resin, filtered, and concentrated. The crude product was dissolved in a mixed solution of tetrahydrofuran / tert-butanol / water (v / v / v = 1:2:2), and Pd / C (10%) and Pd(OH)₂ / C (50%), with an equal mass to that of the starting material 20, were added. The mixture was stirred overnight under a hydrogen atmosphere. After mass spectrometry confirmed the reaction was complete, the mixture was filtered, and the filtrate was lyophilized. The crude product was purified by C18 column chromatography (elution: water) to give the main-chain trisaccharide compound 36 in 44% yield (purity > 95%). 1 H NMR(600MHz,D2O)δ5.14(t,J=1.9Hz,1H),4.88(dd,J=3.9,2.6Hz,1H),4.83(t,J=3.4Hz,1H),4.66(s,56H),4. 54(dd,J=2.8,1.5Hz,1H),4.28-4.24(m,1H),4.18-4.15(m,1H),4.11(dd,J=2.7,1.4Hz,1H),4.01-3.98(m,1H) ,3.93-3.88(m,1H),3.87-3.82(m,1H),3.82-3.76(m,2H),3.72-3.64(m,2H),3.60–3.54(m,1H),3.49–3.40(m ,1H),3.32-3.26(m,1H),2.90-2.83(m,2H),1.59–1.46(m,4H),1.38–1.27(m,2H),1.13(dd,J=6.3,3.1Hz,3H). 13 C NMR (150MHz, D2O) δ 175.93, 174.70, 160.27, 98.36, 98.30, 97.25, 77.13, 75.81, 71.96, 71.85, 70.77, 70.57, 69.34, 68.97, 67.94, 67.85, 39.34, 28.02, 26.29, 22.33, 16.53. HRMS (ESI) calculated values: C 23 H 39 NO 17 H + [M+H] + 602.2291, measured value: 602.2292.

[0371] Step 2: Dissolve compound 23 in a mixed solution of 1,4-dioxane / methanol / 1M sodium hydroxide (v / v / v = 3:1:1), maintain pH 10–11, and stir overnight at room temperature. After mass spectrometry analysis shows complete removal of all ester groups from the raw material, add Amberlite IR120 H... +The reaction solution was neutralized with resin, filtered, and concentrated. The crude product was dissolved in a mixed solution of tetrahydrofuran / tert-butanol / water (v / v / v = 1:2:2), and Pd / C (10%) and Pd(OH)₂ / C (50%), in equal mass to that of starting material 23, were added. The mixture was stirred overnight under a hydrogen atmosphere. After mass spectrometry confirmed the reaction was complete, the mixture was filtered, and the filtrate was lyophilized. The crude product was purified by C18 column chromatography (elution: water) to give the main-chain heptasaccharide compound 37 in 51% yield (purity > 95%). 1 H NMR(800MHz,D2O)δ5.17–5.15(m,3H),4.93(d,J=3.8Hz,3H),4.87(d,J=3.9Hz,1H),4.69–4.65(m,4H),4. 34(dd,J=3.1,1.6Hz,2H),4.31–4.30(m,1H),4.22–4.20(m,2H),4.06-4.01(m,5H),3.95(dd,J=10.3,3.1H z,1H),3.91-3.86(m,1H),3.84–3.78(m,6H),3.75-3.71(m,1H),3.69–3.59(m,4H),3.51-3.46(m,1H),3. 36-3.33(m,3H),2.90(t,J=7.5Hz,2H),1.62–1.51(m,4H),1.40–1.30(m,2H),1.16(dd,J=6.4,1.7Hz,9H). 13 C NMR (200MHz, D₂O) δ 174.17, 98.58 (d, J = 15.3Hz), 98.32, 97.49, 97.39, 77.35 (d, J = 14.0Hz), 76.05 (d, J = 23.9Hz), 71.74 (d, J = 20.6Hz), 70.87, 70.59, 70.37, 69.94, 69.23 (d, J = 22.9Hz), 69.07 (d, J = 5.7Hz), 68.01, 67.90–67.59 (m), 39.33, 28.01, 26.31, 22.34, 16.51. HRMS (ESI) calculated values: C 47 H 75 NO 37 H + [M+H] + 1246.4091, Measured value: 1246.4093

[0372] Test Example 1: Western blot assay to detect the inhibitory effect of cellular-level compounds on TGFβ-induced activation of hepatic stellate cells.

[0373] Human hepatic stellate cell line LX-2 (purchased from Haling Biotechnology) was cultured in 1640 medium (purchased from Thermo Fisher) containing 10% fetal bovine serum FBS (purchased from Gibco, USA) and 1% penicillin antibiotics (purchased from Meilun Biotechnology). The incubation temperature was 37℃ and the CO2 content was 5%. When the cell density reached 80%, LX-2 cells were digested and seeded into 6-well plates. After 24 hours of adherent culture, the original medium was removed, and TGF-β (10 ng / mL, purchased from Sinocare Biotechnology Co., Ltd.) was added, along with trisaccharide compound 1 (100 μM), main chain decasaccharide 26 (100 μM), side chain compounds R1 (i.e., the deprotected product of compound 28 in Example 5), R2 (i.e., the deprotected product of compound 31 in Example 5), and R3 (i.e., the deprotected product of compound 33 in Example 5) (100 μM), and the positive compound OCA (obeticholic acid). The cells were treated with 10 μM of citric acid (purchased from Sigma-Aldrich) medium for 48 hours, followed by cell protein extraction and Western blotting experiments [the main antibodies used in the experiment were: anti-FN1 (#26836S, Cell Signaling Technology), anti-collagen (bs-10423R, Bioss), anti-αSMA (A17910, ABclonal), and anti-GAPDH antibody (AC002, ABclonal)] to examine the expression of liver fibrosis marker proteins by compounds 1 (63 glucose), 26 (10 glucose), and side chain compounds R1-R3 (as shown in Figure 1). After the addition of TGFβ, FN1, Collagen I, and αSMA were all upregulated, indicating successful activation of hepatic stellate cells and successful establishment of the liver fibrosis cell model. However, the expression of FN1, Collagen I, and αSMA was not detected after the addition of compounds 1, 26, and side chain compounds R1-R3.

[0374] The results showed that compound 1 of the hexatrisaccharide and decasaccharide 26 could effectively inhibit TGF-β-induced activation of hepatic stellate cells and reduce the expression of TGF-β-induced fibrosis markers α-SMA, Fibronectin, and Collagen I, which may serve as potential therapeutic agents for liver fibrosis or complications caused by liver fibrosis. Furthermore, the results also showed that the main chain portion of RG-I type pectin-lycium barbarum 63-saccharide (i.e., main chain decasaccharide 26) exhibited superior activity compared to RG-I type pectin-lycium barbarum 63-saccharide.

[0375] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A polysaccharide composition, characterized in that, The polysaccharide composition contains a polysaccharide or a salt thereof as shown in Formula I (preferably, the polysaccharide composition contains ≥90% of a polysaccharide or a salt thereof as shown in Formula I, based on the total mass of the polysaccharide composition); in, R 1 is hydrogen, galactose, galactan or rhamnogalactan; R 2 is hydrogen, arabinose or arabinan; R 3 is hydrogen, galactose, galactan or arabinogalactan; R is hydrogen or -M 1 -(M 2 ) o -M 3 Where o is 0 or an integer between 1 and 30, M 1 For none or C 1-2 Alkylene, M 2 Each is independently a divalent group selected from the following group: -OC 1-2 Alkylene-, C 1-2 Alkylene and M 3 Selected from the following groups: hydrogen, -NH2, -NH(C) 1-4 alkyl) and -NH(C 1-4 Alkyl)2.

2. The polysaccharide composition according to claim 1, characterized in that, The polysaccharide shown in Formula I is polysaccharide 26 as shown below: Alternatively, the polysaccharide shown in Formula I is polysaccharide 1 as shown below:

3. A polysaccharide composition, characterized in that, The polysaccharide composition contains a polysaccharide or a salt thereof as shown in Formula II (preferably, the polysaccharide composition contains ≥90% of a polysaccharide or a salt thereof as shown in Formula I, based on the total mass of the polysaccharide composition); in, R a To hydrogen or R b To hydrogen or Furthermore, R a and R b At least one of them is not hydrogen; R represents hydrogen or -M1-(M2) o -M3: where o is 0 or an integer from 1 to 30, and M1 is none or C. 1-2 The alkylene groups M2 are each independently a divalent group selected from the group consisting of: -OC 1-2 Alkylene-, C 1-2 Alkyl groups and M3 are selected from the following group: hydrogen, -NH2, -NH(C 1-4 alkyl) and -NH(C 1-4 Alkyl)2; n can be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

4. The polysaccharide composition of claim 3, wherein, The polysaccharide shown in Formula II is selected from polysaccharide 26, polysaccharide 36 and polysaccharide 37 as shown below:

5. A pharmaceutical or health product composition, characterized in that, The pharmaceutical composition comprises: (a) the polysaccharide composition according to any one of claims 1-4; and (b) A pharmaceutically or health product-acceptable carrier.

6. Use of the polysaccharide composition according to any one of claims 1-4 in the preparation of a medicament or health product for treating or preventing liver fibrosis or complications caused by liver fibrosis.

7. A polysaccharide intermediate, characterized in that, The intermediate is shown in Formula III; in, R I a To Hydrogen, R I 1 R I 8 or R I b To -OR I 1 、R I 2 or R I c R I 1 R I 3 ; each R I 1 are identical alcohol hydroxyl protecting groups; R I 2 is a -O- alcohol hydroxyl protecting group or leaving group; Each R I 3 Each independently is different from R I 1 The alcohol hydroxyl protecting group; each R I 4 are identical carboxyl protecting groups; R I 5 Selected from the following groups: hydrogen, alcohol hydroxyl protecting group, -M I 1 -(M I 2 ) o -M I 3 Where o is 0 or an integer between 1 and 30, M 1 For none or C 1-2 Alkylene, M 2 Each is independently a divalent group selected from the following group: -OC 1-2 Alkylene-, C 1-2 Alkylene and M 3 It is hydrogen or -N(R) P )2; where R P C 1-4 Alkyl or amino protecting groups; R I 8 is hydrogen or an alcohol hydroxyl protecting group different from R I 1 ; R I 7 is an alcohol hydroxyl protecting group.

8. The polysaccharide intermediate as described in claim 7, characterized in that, (a) The polysaccharide intermediate is shown in Formula III-1; Among them, R I 1 R I 4 R I 5 R I 7 and R I c As defined in Equation III; preferably, where R I 1 For Bn, R I 4 For Bn, R I 5 -(CH2) s -N(Bn)Cbz, where s is an integer from 1 to 10, R I 7 For Bz; each R I c The 0, 1, 2, 3, 4 or all of them are R I 3 And the rest of R I c For R I 1 And R I 3 As defined in Equation III; Preferably, the polysaccharide intermediate is as shown in Formula III-1a; Among them, R I 1 R I 4 R I 5 and R I 7 As defined in Equation III-1; R I 31 R I 32 and R I 33 The definition of R is the same as in equation III-1 I 3 Definition; preferably, where R I 1 For Bn, R I 4 For Bn, R I 5 For R I 5 -(CH2) s -N(Bn)Cbz, where s is an integer from 1 to 10, R I 7 For Bz, R I 31 R I 32 and R I 33 Each is an independent protecting group for a different alcohol hydroxyl group, and R I 31 R I 32 and R I 33 Both are different from R I 1 R I 4 and R I 7 ; More preferably, the polysaccharide intermediate is compound 25; or, (b) The polysaccharide intermediate is shown in Formula III-2; Among them, R I 1 R I 2 R I 4 and R I c As defined in Equation III; R I 8 The definition of R in Equation III is the same. I 3 Definition; preferably, where R I 1 For Bn, R I 2 for -O-All, -O-PTFAI, or -O-TCAI, R I 4 For Bn, R I 8 It is an alcohol hydroxyl protecting group, and R I 8 Unlike R I 1 R I 2 R I 4 and R I c R I c For R I 3 or R I 1 And R I 3 As defined in Equation III; Preferably, the polysaccharide intermediate is as shown in formula III-2a or III-2b; Among them, R I 1 R I 2 R I 4 and R I 8 As defined in Equation III-2; R I 31 R I 32 and R I 33 The definition of R is the same as in equation III-2. I 3 Definition; preferably, where R I 1 For Bn, R I 2 for -O-All, -O-PTFAI, or -O-TCAI, R I 4 For Bn, R I 8 R is one of Lev, Nap, and Fmoc. I 32 and R I 33 All are alcohol hydroxyl protecting groups, and R I 32 and R I 33 Both are different from R I 1 R I 2 R I 4 and R I 8 ; More preferably, the polysaccharide intermediate is selected from the group consisting of: compound 7, compound 8, compound 16, and compound 17; or, (c) The polysaccharide intermediate is shown in Formula III-3; Among them, R I 1 R I 2 R I 4 R I 7 and R I c As defined in Equation III; preferably, where R I 1 For Bn, R I 2 for -O-All, -O-PTFAI, or -O-TCAI, R I 4 For Bn, each R I c 0, 1, or all of them are R I 3 And the rest of R I c For R I 1 And R I 3 As defined in Equation III; Preferably, the polysaccharide intermediate is as shown in Formula III-3a; Among them, R I 1 R I 2 R I 4 and R I 7 As defined in Equation III-3, R I 31 The definition of R is the same as in equation III-1 I 3 Definition; preferably, where R I 1 For Bn, R I 2 for -O-All, -O-PTFAI, or -O-TCAI, R I 4 For Bn, R I 31 It is an alcohol hydroxyl protecting group, and R I 31 Both are different from R I 1 R I 2 R I 4 and R I 7 ; More preferably, the polysaccharide intermediate is compound 14 or 15; or, (d) The polysaccharide intermediate is shown in Formula III-4; Among them, R I 1 R I 4 R I 5 and R I c As defined in Equation III; R I 8 The definition of R in equation III is the same. I 3 Definition; preferably, where R I 1 For Bn, R I 4 For Bn, R I 5 -(CH2) s -N(Bn)Cbz, where s is an integer from 1 to 10, R I 8 It is an alcohol hydroxyl protecting group, and R I 8 Unlike R I 1 R I 2 R I 4 and R I c R I c For R I 3 or R I 1 And R I 1 and R I 3 As defined in Equation III; Preferably, the polysaccharide intermediate is as shown in formula III-4a; Among them, R I 1 R I 4 R I 5 and R I 8 As defined in equation III-4; R I 33 As shown in equation III-4, R I 3 The definition of; preferably, where, I 1 For Bn, R I 4 For Bn, R I 5 -(CH2) s -N(Bn)Cbz, where s is an integer from 1 to 10, R I 8 R is one of Lev, Nap, and Fmoc. I 33 It is an alcohol hydroxyl protecting group, and R I 33 Unlike R I 1 R I 2 R I 4 and R I 8 ; More preferably, the polysaccharide intermediate is compound 19; or, (e) the polysaccharide intermediate is as shown in formula III-5; Among them, R I 1 R I 2 R I 4 and R I c As defined in Equation III; R I 8 The definition of R in equation III is the same. I 3 Definition; preferably, where R I 1 For Bn, R I 2 for -O-All, -O-PTFAI, or -O-TCAI, R I 4 For Bn, R I 8 It is an alcohol hydroxyl protecting group, and R I 8 Unlike R I 1 R I 2 R I 4 and R I c R I c 0, 1, or all of them are R I 3 And the rest of R I c For R I 1 And R I 3 As defined in Equation III; Preferably, the polysaccharide intermediate is as shown in formula III-5a; Among them, R I 1 R I 2 R I 4 and R I 8 As defined in Equation III-5; R I 32 The definition of R is the same as in equation III-5. I 3 Definition; preferably, where R I 1 For Bn, R I 2 for -O-All, -O-PTFAI, or -O-TCAI, R I 4 For Bn, R I 8 R is one of Lev, Nap, and Fmoc. I 32 It is an alcohol hydroxyl protecting group, and R I 3 Unlike R I 1 R I 2 R I 4 and R I 8 ; More preferably, said polysaccharide intermediate is compound 21 or 22; or, (f) the polysaccharide intermediate is represented by Formula III-6; Among them, R I 1 R I 4 R I 5 and R I c As defined in Equation III; R I 8 The definition of R in equation III is the same. I 3 Definition; preferably, where R I 1 For Bn, R I 4 For Bn, R I 5 -(CH2) s -N(Bn)Cbz, where s is an integer R from 1 to 10. I 8 It is an alcohol hydroxyl protecting group, and R I 8 Unlike R I 1 R I 4 and R I c Each R I c 0, 1, 2 or all of them are R I 3 And the rest of R I c For R I 1 And R I 3 As defined in Equation III; Preferably, the polysaccharide intermediate is as shown in formula III-6a; Among them, R I 1 R I 4 R I 5 and R I 8 As defined in Equation III-6; R I 32 and R I 33 In formula III-6, R I 3 Definition; preferably, where R I 1 For Bn, R I 4 For Bn, R I 5 -(CH2) s -N(Bn)Cbz, where s is an integer from 1 to 10, R I 7 For Bz, R I 8 R is one of Lev, Nap, and Fmoc. I 32 and R I 33 It is an alcohol hydroxyl protecting group, and R I 32 and R I 33 Unlike R I 1 R I 2 R I 4 and R I 8 ; More preferably, said polysaccharide intermediate is compound 23; 9. A method for synthesizing an a-GalA-(1→2)-Rha glycosidic bond in a disaccharide module as shown in Formula IV, characterized in that comprising the steps of: Among the various types, R I 1 R I 2 R I 4 and R I c As defined in Formula III of claim 6; R I L It is a leaving group; preferably, R I L Selected from the following group: -SC 1-6 Alkyl, -S-unsubstituted or with one or more C 1-6 Alkyl-substituted phenyl, trichloroacetamide imine, tribromoacetamide imine, and trifluoroacetylimine esters; more preferably, R I L -SC 1-6 Alkyl or -S- unsubstituted or with one or more Cs 1-6 Alkyl-substituted phenyl; more preferably, R I L -S- is not replaced or is occupied by one or more Cs 1-6 Alkyl-substituted phenyl groups (e.g., -SPh, -STol); preferably, R I L -STol; (S1) Glycosylate intermediates of formula IV-1 and IV-2 to obtain intermediate IV-3; preferably, the glycosylation reaction is carried out in the presence of AgOTf and / or Ar-SCl and / or TTBP; more preferably, the glycosylation reaction is carried out in the presence of AgOTf, p-TolSCl, TTBP and molecular sieve; (S2) The intermediate of formula IV-3 is subjected to a deprotection reaction to remove the >Si(tBu)2 protecting group, thereby obtaining the intermediate of formula IV-4; preferably, the deprotection reaction is carried out in the presence of a fluoride; more preferably, the deprotection reaction is carried out in the presence of an HF / Py complex; (S3) The intermediate of formula IV-4 is oxidized to obtain the intermediate of formula IV-5 containing an α-GalA-(1→2)-Rha glycosidic bond; preferably, the oxidation reaction is carried out in the presence of an oxidizing agent selected from the group consisting of TEMPO (2,2,6,6-tetramethylpiperidine-nitrogen oxide), BAIB (iodophenyldiacetic acid), or a combination thereof.

10. A method of preparing a polysaccharide composition as claimed in claim 1, characterised in that, Including the following steps: (1) providing an intermediate of Formula III-1a Among them, R I 1 R I 4 R I 5 and R I 7 As defined in Formula III-1 of claim 8; R I 31 R I 32 and R I 33 The definition is the same as R in formula III-1 as claimed in claim 8. I 3 Definition; (2) Make the intermediate R of formula III-1a 32 Positional deprotection; and optionally, the deprotection product is reacted with R. S 1 -R S L The reaction yields branched polysaccharide intermediate 1. (3) R of the branched polysaccharide intermediate 1 obtained in step (2) 31 Positional deprotection; and optionally, the deprotection product is reacted with R. S 2 -R S L The reaction yields branched polysaccharide intermediate 2; (4) R of the branched polysaccharide intermediate 2 obtained in step (3) 33 Positional deprotection; and optionally, the deprotection product is reacted with R. S 3 -R S L The reaction yields branched polysaccharide intermediate 3; (5) Deprotection reaction is performed on branched polysaccharide intermediate 3 to obtain the polysaccharide composition as described in the first aspect; Among them, R S L R is a leaving group; S 1 R S 2 and R S 3 R, where the hydroxyl group is protected. 1 R 2 and R 3 ;R 1 R 2 and R 3 As defined in Equation I.

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