Saponin conjugates and vaccines or pharmaceutical compositions containing same

Abstract

The present invention relates to novel compounds in which a lipophilic moiety, such as a lipid, fatty acid, polyethylene glycol, or terpene, is covalently attached to a non-acylated or de-acylated triterpene saponin via the carboxyl group present in the 3-O-glucuronic acid of the triterpene saponin. The attachment of a lipophilic moiety to the 3-O-glucuronic acid of a saponin, such as Quillaja de-acylated saponin, lucyoside P, or saponins from Gypsophila, Saponaria, and Acanthophyllum, enhances the adjuvant effect on humoral and cell-mediated immunity. Furthermore, the saponin analogs obtained by attaching a lipophilic moiety to the 3-O-glucuronic acid residue of non-acylated or de-acylated saponins are easier to purify, less toxic, and chemically stable than the original saponins, and have comparable or even superior adjuvant properties.

Description

[Technical field]

[0001] The present invention relates to saponin conjugates, their synthesis, and intermediates thereof. The present invention also provides pharmaceutical compositions comprising the saponin conjugates of the invention, and methods of using said saponin conjugates or compositions in the treatment of infectious diseases, cancer, and immunological disorders. [Background technology]

[0002] Adjuvants have proven their effectiveness in current vaccine regimens. The current remaining challenge is that for therapeutic vaccines, the antigen-adjuvant combination must be effective in providing both humoral and cellular immunity to treat complex diseases such as HIV, malaria, tuberculosis and cancer. Providing pathogen-specific T cell responses is fundamental for developing new therapeutic vaccines, where adjuvants play a role. However, few adjuvants are potent enough to induce cellular immunity and non-toxic for clinical use.

[0003] Quillaja saponaria (or Q. saponin) is a triterpene glycoside isolated from the Chilean Quillaja Saponaria Molina. Q. saponin is a potent stimulator of mucus production in the respiratory tract and causes inflammation in the digestive tract. Four major triterpenoid glucosides were isolated from Quillaja saponaria extracts and identified as QS-7, QS-17, QS-18, and QS-21 (Quillaja saponin fractions-7, 17, 18, and 21). [4]Their structures were subsequently assigned as shown below. All these saponins share the same triterpene backbone quilacic acid and adjacent branched trisaccharide β-D-Gal-(1→2)-[β-D-Xyl-(1→3)]-β-D-GlcA at the 3-O position. QS-21 is a stereochemically rich fatty acyl chain with Fuc and fucose-linked 4-O-acyl at the 28-O position containing a linear tetrasaccharide moiety β-D-Apif / Xylp-(1→3)-β-D-Xyl-(1→4)-α-L-Rha-(1→2)-β-D-. The structures of QS-21 and its purified analogues are shown. [ka] TIFF0007680966000002.tif36143 Apif: apiofuranos; Xylp: xylopyranose; Glcp: glucopyranose; Rhap: Rhamnopyranose

[0004] QS-21 has been established as a promising adjuvant for immune response enhancement and dose sparing due to its potency and favorable toxicity profile in hundreds of recent and ongoing vaccine clinical trials (malaria, herpes, Alzheimer's disease, HIV-1, melanoma, breast cancer, small cell lung cancer, prostate cancer, etc.). However, it remains problematic due to four major issues: dose-limiting toxicity, inadequate stability, poor understanding of its molecular mechanism of action, and limited availability of high-quality products.

[0005] GPI-0100 is a semi-synthetic saponin mixture derived from Quillaja extract. Crude bark extract was treated under mild basic hydrolysis and then coupled with aliphatic dodeacyl chains via hydrolytically stable amide bonds to obtain GPI-0100. This modification certainly gave these molecules more resistance at high temperatures. Furthermore, the toxicity inherent to Quillaja extract was uncoupled from its immunological stimulating ability. However, its adjuvant activity was reduced. Thus, there remains a need for adjuvants that enhance cellular immunity and reduce toxicity. The present invention develops a new generation of saponin-based adjuvants with improved efficacy in cellular immunity, which are more suitable for combination with therapeutic vaccines than existing ones. Summary of the Invention

[0006] The present invention relates to novel compounds referred to herein as saponin conjugates.

[0007] In one aspect, the present invention provides a saponin conjugate of formula (I) or a pharma- ceutically acceptable salt thereof: [ka] (I) [where: TIFF0007680966000004.tif39 is a single or double bond, W is a methyl group (Me), -CHO, TIFF0007680966000005.tif1630,-CH 2 OR 1 ,-C(O)R x , or C.H. 2 OR x and V is hydrogen or -OR 1 and Y is CH 2 , -O-, -S-, -NR-, or -NH-; Q is CH 2 , C=O, C=N-OH, or C=N-OMe; X is CH 2, -O-, -NR-, -NH-(C=O)-, -S-, or O-(C=O)-; R is a cyclic or acyclic, optionally substituted moiety selected from the group consisting of acyl, aliphatic, heteroaliphatic, aryl, arylaliphatic, cycloaliphatic, heterocyclic aliphatic, heteroarylaliphatic, alkyloxyaliphatic, and aryloxyaliphatic, or C 1 ~C 18 an optionally substituted moiety selected from the group consisting of aliphatic, 5-10 membered arylaliphatic, 5-10 membered heteroarylaliphatic having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and 4-7 membered heterocyclylaliphatic having 1-2 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur; R 1 are independently hydrogen, an oxygen protecting group selected from the group consisting of alkyl ether, benzyl ether, silyl ether, acetal, ketal, ester, carbamate and carbonate, or a carbohydrate having the monosaccharide structure set forth below, such as glucose, mannose, galactose, N-acetylglucosamine, N-acetylgalactosamine, altrose, allose, fucose, rhamnose, etc. [ka] (where: a, b, and c, independently in each occurrence, are 0 or 1; R 0 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R a , R b , R c , and R d is independently in each occurrence hydrogen, halogen, OH, OR, OR x and R xis independently in each occurrence hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates. Z is hydrogen, a cyclic or acyclic, optionally substituted moiety selected from the group consisting of acyl, aliphatic, heteroaliphatic, aryl, arylalkyl, heterocyclyl and heteroaryl, or a carbohydrate domain having the structure: [ka] Where: R 2 is, at each occurrence, H or a carbohydrate domain having the structure: [ka] Where: a, b and c are independently in each occurrence 0, 1, or 2; d is an integer of 1 to 5, and the structures enclosed in the brackets d may be the same or different, provided that the structures enclosed in the brackets d represent furanose or pyranose moieties, and the sum of b and c is 1 or 2; R 0 is hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R a , R b , R c and R d is independently in each occurrence hydrogen, halogen, OH, OR, OR x , N.R. 2 , NHCOR or acyl, C 1 ~C 10 aliphatic, C 1 ~C 6An optionally substituted group selected from the group consisting of heteroaliphatic, 6- to 10-membered aryl, arylaliphatic, 5- to 10-membered heteroaryl having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and 4- to 7-membered heterocyclyl having 1 to 2 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. R 3 is hydrogen, halogen, OH, OR x and R 4 is hydrogen, halogen, CH 2 OR x or acyl, C 1 ~C 10 an optionally substituted group selected from the group consisting of aliphatic R x is independently in each occurrence hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates.

[0008] The present invention encompasses the recognition that the clinical use of GPI-0100 as a mixture of adjuvants is limited due to its structural complexity and difficulty in isolating it in a pure form.The present invention provides compounds that are analogs of GPI-0100.

[0009] According to another aspect, the compounds of the present invention are shown to be useful as adjuvants.Therefore, in certain embodiments, a vaccine is provided that comprises one or more bacterial antigens, virus-associated antigens, protozoa-associated antigens or tumor-associated antigens and one or more compounds of the present invention.In certain embodiments, one or more antigens are non-covalently associated with a pharmaceutically acceptable excipient.In some embodiments, one or more antigens are covalently bound to a pharmaceutically acceptable excipient.

[0010] In another aspect, the present invention provides a method of enhancing an immune response to an antigen comprising administering to a subject an effective amount of a provided vaccine to enhance the immune response to the antigen in said subject.

[0011] In another embodiment, the present invention provides saponin agents that induce immune responses towards humoral and cellular immunity.

[0012] In another embodiment, the present invention provides a method of stimulating or enhancing cytokine production in a subject, the method comprising, inter alia, administering to the subject any one of the compounds of the present invention, whereby immune cells secrete cytokines.

[0013] In another aspect, the invention provides a method of vaccinating a subject, comprising administering to the subject a provided vaccine. In some embodiments, the subject is a human. In some embodiments, the vaccine is administered orally. In other embodiments, the vaccine is administered intramuscularly. In other embodiments, the vaccine is administered subcutaneously. In certain embodiments, the dose of the adjuvant compound is 10-1000 μg. In certain embodiments, the dose of the adjuvant compound is 500-1000 μg. In certain embodiments, the dose of the adjuvant compound is 100-500 μg. In certain embodiments, the dose of the adjuvant compound is 50-250 μg. In certain embodiments, the dose of the adjuvant compound is 50-500 μg. In certain embodiments, the dose of the adjuvant compound is 250-500 μg. The antigen against which the subject is vaccinated may be a cancer-associated antigen, a bacterial antigen, a virus-associated antigen, a protozoan-associated antigen, or a self-antigen.

[0014] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention and a pharma- ceutical acceptable excipient. In certain embodiments, the pharmaceutical composition is a vaccine comprising an antigen and an adjuvant of the present invention.

[0015] In another aspect, the invention provides kits comprising pharmaceutical compositions of the compounds of the invention. In some embodiments, the kits comprise prescribing information. In some embodiments, such kits comprise a combination of the adjuvant compounds of the invention and another immunotherapeutic agent (e.g., vaccine, antibody). These agents may be packaged separately or together. The kits optionally comprise instructions for prescribing the drug. In certain embodiments, the kits comprise multiple doses of each agent. The kits may comprise a sufficient amount of each component to treat a subject for one week, two weeks, three weeks, four weeks, or several months. In certain embodiments, the kits comprise one cycle of immunotherapy. In certain embodiments, the kits comprise a sufficient amount of the pharmaceutical composition to immunize a subject against an antigen for an extended period of time.

[0016] In one embodiment, the present invention provides a process for preparing a compound represented by the structure of formula (II) or a pharma- ceutically acceptable salt thereof. [ka] (II) [where: TIFF0007680966000010.tif39 is a single bond or a double bond, W is Me, -CHO, TIFF0007680966000011.tif1732,-CH 2 OR 1 , -C(O)R, or CH 2 OR x and V is hydrogen or -OR 1 and Y is CH 2 , -O-, -S-, -NR-, -NH-, or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; Q is CH 2 , C=O, C=N-OH, or C=N-OMe; X is CH 2, -O-, -NR-, -NH-(C=O)-, -S-, or O-(C=O)-; R 1 is independently hydrogen, an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates and carbonates, or a carbohydrate having the structure: [ka] (where: a, b, and c are independently in each occurrence 0 or 1; R 0 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R a , R b , R c , and R d is independently in each occurrence hydrogen, halogen, OH, OR, OR x and R x is independently in each occurrence hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates. R is a cyclic or acyclic, optionally substituted moiety selected from the group consisting of acyl, aliphatic, heteroaliphatic, aryl, arylaliphatic, cycloaliphatic, heterocyclic aliphatic, heteroarylaliphatic, aryloxyaliphatic, and aryloxyaliphatic, or C 1-18 an optionally substituted moiety selected from the group consisting of aliphatic, 5-10 membered arylaliphatic, 5-10 membered heteroarylaliphatic having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and 4-7 membered heterocyclylaliphatic having 1-2 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur.

[0017] In one embodiment of the present invention, the compound of formula (II) may be obtained by a process comprising, inter alia, the following steps: A compound represented by the formula (III) is reacted with a compound represented by the formula (IV) or a pharma- ceutically acceptable salt thereof. [ka] (III) R 1 is independently hydrogen, an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates and carbonates, or a carbohydrate having the structure: [ka] , Where: a, b, and c, independently in each occurrence, are 0 or 1; R 0 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R a , R b , R c , and R d is independently in each occurrence hydrogen, halogen, OH, OR, OR x and R x is independently in each occurrence hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; LG is a leaving group, inter alia TIFF0007680966000015.tif2036 or It could also be TIFF0007680966000016.tif2035. [ka] (IV) Where: TIFF0007680966000018.tif310 is a single bond or a double bond, W is Me, -CHO, TIFF0007680966000019.tif1731,-CH 2 OR or -C(O)R; V is hydrogen or -OR 1 and Y is CH 2 , -O-, -NR-, -NH-, or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R, R x or R 1 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates.

[0018] In one embodiment of the present invention, a type of compound of formula (I) can be obtained by a process comprising, inter alia, the following steps: The compound of formula (II) is reacted with a compound of formula (V) or a pharma- ceutically acceptable salt thereof. [ka] (V), Where: PG and PG 1 is an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R 1 is independently hydrogen, an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates and carbonates, or a carbohydrate having the structure: [ka] Where: a, b, and c, independently in each occurrence, are 0 or 1; R 0 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R a , R b , R c , and R d is independently in each occurrence hydrogen, halogen, OH, OR, OR x and R x is independently in each occurrence hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates. R 2 are independently hydrogen, halogen, CH 2 OH, or a lower alkyl group, LG is a leaving group, inter alia TIFF0007680966000022.tif2036 or It could also be TIFF0007680966000023.tif2035. (definition) As used herein, the following definitions apply unless otherwise stated.

[0019] "Stereoisomer" or "stereoisomers" refers to a compound that differs in chirality at one or more stereocenters. Stereoisomers include enantiomers and diastereomers.

[0020] "Subject" refers to mammals, including human and non-human mammals.

[0021] As used herein, the term "aliphatic" or "aliphatic group" or "aliphatic moiety" refers to a linear (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more units of unsaturation, or a non-aromatic monocyclic or bicyclic hydrocarbon (also referred to herein as "carbocycle," "aliphatic," or "cycloalkyl") that is fully saturated or contains one or more units of unsaturation and has a single point of attachment to the remainder of the molecule. Unless otherwise specified, an aliphatic group contains aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-12 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-11 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-10 aliphatic carbon atoms. In still other embodiments, an aliphatic group contains 1-9 aliphatic carbon atoms, and in still other embodiments, an aliphatic group contains 1-8 aliphatic carbon atoms. In still other embodiments, an aliphatic group contains 1-7 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-6 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms. In other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms.

[0022] In some embodiments, cycloaliphatic (also referred to as "carbocycle" or "cycloalkyl") refers to a non-aromatic monocyclic C-C hydrocarbon that is fully saturated or contains one or more units of unsaturation and has a single point of attachment to the remainder of the molecule. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and complexes thereof (e.g., (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl), and the like.

[0023] The term "heteroatom" means one or more oxygen, sulfur, nitrogen, phosphorus, or silicon (including any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen, or a substitutable nitrogen of a heterocycle, such as N (e.g., in 3,4-dihydro-2H-pyrrolyl), NH (e.g., in pyrrolidinyl), or NR'' (e.g., in N-substituted pyrrolidinyl)).

[0024] As used herein, the term "unsaturated" means a moiety that has one or more double bonds.

[0025] The term "halogen" means F, Cl, Br, or I.

[0026] The term "acyl" used alone or as part of a larger moiety means a group formed by removal of the hydroxy group from a carboxylic acid.

[0027] The terms "arylkyl" and "arylaliphatic" are used interchangeably and refer to an aliphatic group in which a hydrogen atom is replaced with an aryl group, including, but not limited to, phenyl, biphenyl, naphthyl, cinnamyl, and dihydrocinnamyl.

[0028] The term "aryl" is used alone or as part of a larger moiety such as "arylaliphatic" or "heteroarylaliphatic."

[0029] The term "aryloxyaliphatic" (or "aralkoxy, or arylkoxy, or "aryloxyalkyl") refers to a monocyclic or bicyclic ring system having a total of 5 to 14 ring members, where at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. The term "aryl" can be used interchangeably with the term "aryl ring."

[0030] In certain embodiments of the invention, "aryl" refers to an aromatic ring system, which may bear one or more substituents, including, but not limited to, benzyl, phenyl, biphenyl, naphthyl, anthracyl, etc. Also included within the scope of the term "aryl" as used herein are groups in which an aromatic ring is fused with zero or more non-aromatic rings, such as indanyl, phthalimidyl, naphthymidyl, phenanthridinyl, or tetrahydronaphthyl.

[0031] The term "heteroaryl" used alone or as part of a larger moiety, for example, "heteroaryloxy" or "heteroarylaliphatic" or "heteroarylalkyl" refers to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; groups having 6, 10, or 14 electrons shared in a cyclic array; and groups having 1 to 5 heteroatoms in addition to the carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl.

[0032] The terms "heteroaryl" and "heteroar" as used herein also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, and the attachment point is on the heteroaromatic ring.Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and 2H-pyrido[2,3-b]-1,4-oxazin-3(4H)-one.Heteroaryl groups may be monocyclic or bicyclic. The term "heteroaryl" can be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which includes rings that are optionally substituted. The terms "heteroarylaliphatic" and "heteroarylalkyl" refer to an aliphatic group substituted by a heteroaryl moiety, where the aliphatic and heteroaryl moieties independently are optionally substituted.

[0033] As used herein, the term "heteroaliphatic" refers to an aliphatic group in which one or two carbon atoms are independently replaced by one or more of oxygen, sulfur, nitrogen, or phosphorus. Heteroaliphatic groups can be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and include "heterocycle," "heterocyclyl," "heterocycloaliphatic," or "heterocyclic" groups.

[0034] As used herein, the terms "heterocycle", "heterocyclyl" and "heterocycle" are used interchangeably and refer to a saturated or partially unsaturated stable 5-7 membered monocyclic or 7-10 membered bicyclic heterocyclyl moiety having, in addition to carbon atoms, one or more, preferably one to four heteroatoms as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen can be N (e.g., in 3,4-dihydro-2H-pyrrolyl), NH (e.g., in pyrrolidinyl), or . + It may also be NR (eg, as in N-substituted pyrrolidinyl).

[0035] A heterocycle can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any ring atom can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl.

[0036] The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," and "heterocyclic moiety" are used interchangeably herein and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings, such as, for example, indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. Heterocyclic groups may be monocyclic or bicyclic.

[0037] The term "heterocyclylaliphatic" refers to a heterocycle-substituted alkyl group, in which the aliphatic and heterocycle portions independently are optionally substituted.

[0038] As used herein, the term "partially unsaturated" refers to a ring moiety that contains at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings having multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties as defined herein.

[0039] In another aspect, the present invention provides a "pharmaceutical acceptable" composition comprising a therapeutically effective amount of one or more compounds described herein, formulated together with one or more pharma- ceutically acceptable carriers (additives) and / or diluents. As described in detail, the pharmaceutical compositions of the present invention may be specially formulated for administration in solid or liquid form, including those adapted for "oral administration, e.g., drench (aqueous or non-aqueous solution or suspension), tablet (e.g., buccal, sublingual, and systemic absorption), bolus, powder, granule, paste for application to the tongue", "parenteral administration, e.g., sterile solution or suspension or sustained release formulation by intramuscular, subcutaneous, intravenous, or epidural injection", "topical application, e.g., cream, ointment, or sustained release spray or patch applied to the lung, skin, or oral cavity", "vaginal or rectal application, e.g., pessary, cream, or foam", "sublingual", "ocular", "transdermal", or "nasal, pulmonary, and other mucosal surface".

[0040] The term "pharmacologically acceptable" is used herein to refer to such compounds, compositions, materials, and / or dosage forms, which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings or animals without undue toxicity, irritation, allergic response, or other problem or complication, and are commensurate with a reasonable benefit / risk ratio.

[0041] The term "pharmaceutical acceptable carrier" as used herein means a pharmaceutical acceptable material, composition, or vehicle, such as a liquid or solid filler, excipient, diluent, or solvent encapsulating material that is involved in the transport or delivery of the subject compound from one part of the body to another part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that can be used as pharma- ceutically acceptable carriers include sugars such as glucose, lactose, sucrose, etc.; starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, etc.; powdered tragacanth; malt, gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate, agar, and buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water, isotonic saline: Ringer's solution: ethyl alcohol; pH buffers; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic compatible substances used in the formulation.

[0042] The term "pharmaceutically acceptable salts" as used herein refers to salts that are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, or allergic reaction, within the scope of sound medical judgment, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al., J. Pharmaceutical Sciences, 1977, 66, 1-19, provide a detailed description of pharmaceutically acceptable salts, which are incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed with amino groups and inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, and malonic acid, or by using other methods used in the art, such as ion exchange. Other pharma- ceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, and the like. These salts include but are not limited to carboxylate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, and valerate.

[0043] In other cases, the compounds of the present invention may contain one or more acidic functional groups, which can form pharma- ceutically acceptable salts with pharma-ceutically acceptable bases. In these cases, the term "pharma-ceutically acceptable salts" refers to the relatively non-toxic inorganic and organic base addition salts of the compounds of the present invention. These salts can likewise be prepared in situ during the administration vehicle or dosage form manufacturing process, or by separately reacting the purified compound in free acid form with a suitable base (e.g., hydroxide, carbonate or bicarbonate of a pharma-ceutically acceptable metal cation) with ammonia, or a pharma-ceutically acceptable organic primary, secondary, tertiary, or quaternary amine, and the like. Salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N(C 1-4 Representative pharmaceutically acceptable salts include base addition salts, such as ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl and aryl sulfonates, and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like.

[0044] Unless otherwise stated, structures depicted herein also include all isomers, e.g., enantiomers, diastereomers, and geometric conformational forms of the structure, e.g., R and S configurations of each stereocenter, Z and E double bond isomers, and Z and E conformational isomers. Thus, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the compounds of the invention are within the scope of the invention. Unless otherwise stated, all tautomeric forms of the compounds of the invention are within the scope of the invention.

[0045] The compounds provided may contain one or more sugar moieties. Unless otherwise specified, both D and L configurations and mixtures thereof are within the scope of the present invention. Unless otherwise specified, both C-linked and S-linked embodiments, as well as mixtures thereof, are contemplated by the present invention.

[0046] Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, the replacement of hydrogen by deuterium or tritium, or 13 C or 14 Any compound having a structure of the invention that includes a carbon substitution with a C-rich carbon is within the scope of the invention. Such compounds are useful, for example, as analytical tools, probes in biological assays, or as therapeutic agents according to the invention.

[0047] According to an embodiment of the present invention, the term "protecting group" as used herein refers to a temporary modification of a potentially reactive functional group that protects it from undesired chemical transformations. Examples of such protecting groups include esters of carboxylic acids, silyl ethers of alcohols, and acetals and ketals of aldehydes and ketones, respectively. Of course, other suitable protecting groups can also be used. Further, various protecting groups are described by Greene and Wuts (supra).

[0048] In one embodiment of the present invention, the protecting group may be, inter alia, a hydroxy protecting group. In one embodiment of the present invention, the hydroxy protecting group may be, inter alia, an alkyl, aryl, aralkyl, silyl or acyl radical. In another embodiment, the protecting group may be, inter alia, a trimethylsilyl, triethylsilyl, t-butyldimethylsilyl (TBS), triisopropylsilyl (TIPS) or t-butyldiphenylsilyl. Of course, other suitable protecting groups may be used.

[0049] In one embodiment, the aralkyl may be unsubstituted or substituted. In another embodiment, the aralkyl may be, inter alia, arylmethyl. In another embodiment, the protecting group may be, inter alia, benzyl. In another embodiment, the protecting group may be, inter alia, methoxybenzyl. In another embodiment, the methoxybenzyl may be, inter alia, paramethoxybenzyl.

[0050] In one embodiment of the present invention, the protecting group may be, inter alia, an amino protecting group. In one embodiment of the present invention, the amino protecting group may be, inter alia, a carbamate, an amide or an N-sulfonyl amide. In another embodiment, the amino protecting group may be, inter alia, a benzyloxycarbonyl (Cbz), a 9-fluorenylmethyloxycarbonyl (Fmoc), a t-butyloxycarbonyl (tBoc), a biphenylisopropyloxycarbonyl, a t-amyloxycarbonyl, an isobornyloxycarbonyl, an α-dimethyl-3,5-dimethoxybenzyloxycarbonyl or a 2-cyano-t-butyloxycarbonyl.

[0051] Furthermore, in one embodiment, the present invention provides a method for stimulating, inhibiting, suppressing or modulating an immune response in a subject, which may comprise, inter alia, administering to the subject any one of the compounds of the present invention or any combination thereof.

[0052] Furthermore, in one embodiment, the present invention provides a method for stimulating, inhibiting, suppressing or modulating an immune response in a subject, the method comprising administering to the subject a pharmaceutical composition comprising, inter alia, any one of the compounds of the present invention, or any combination thereof, and one or more pharma- ceutically acceptable excipients.

[0053] Further, in one embodiment, a "pharmaceutical composition" may refer to a therapeutically effective amount of one or more compounds of the present invention and suitable excipients and / or carriers useful for stimulating, inhibiting, suppressing or modulating an immune response in a subject.

[0054] In one embodiment, a "therapeutically effective amount" may refer to an amount that provides a therapeutic effect for a given condition and administration regimen. In one embodiment, the composition may be administered by any method known in the art.

[0055] As described herein, the compounds of the invention may include "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when one or more positions in any given structure may be substituted with one or more substituents selected from a specified group, the substituents at all positions may be the same or different. Combinations of substituents envisioned by the present invention are preferably those that result in the formation of stable or chemically feasible compounds.

[0056] As used herein, the terms "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0057] As used herein, the terms "systemic administration," "administered peripherally," "peripheral administration," and "administered peripherally" refer to administration of a compound, drug, or other substance other than by direct administration into the central nervous system and which is subject to metabolic and other similar processes as it enters the patient's system, e.g., subcutaneous administration.

[0058] The term "pure" refers to a compound that is substantially free of related non-target structures or chemical precursors (if chemically synthesized). This quality may be measured or expressed as "purity." In some embodiments, the target compound has less than about 30%, 20%, 10%, 5%, 2%, 1%, 0.5%, and 0.1% of non-target structures or chemical precursors.

[0059] The term "carbohydrate" refers to a sugar or a polymer of sugar. The terms "sugar", "polysaccharide", "carbohydrate" and "oligosaccharide" may be used interchangeably. Most carbohydrates are aldehydes or ketones with many hydroxyl groups, usually one for each carbon atom of the molecule. Carbohydrates are generally represented by the molecular formula C n H 2n O n The carbohydrates may be monosaccharides, disaccharides, trisaccharides, oligosaccharides, or polysaccharides. The most basic carbohydrates are monosaccharides, such as glucose, galactose, sucrose, ribose, mannose, arabinose, xylose, and fructose. Disaccharides are two linked monosaccharides. Exemplary disaccharides include sucrose, lactose, cellobiose, and maltose. Typically, oligosaccharides contain 3-6 monosaccharide units (e.g., raffinose, stachyose), and polysaccharides contain 6 or more monosaccharide units. Exemplary polysaccharides include starch, glycogen, and cellulose. Carbohydrates may also contain modified sugar units, such as 2'-deoxyribose, where the hydroxyl group has been removed, 2'-fluororibose, where the hydroxyl group has been replaced with a fluorine, or N-acetylglucosamine, which is a nitrogen-containing form of glucose. (e.g., 2'-fluororibose, deoxyribose, and hexose). Carbohydrates may exist in many different forms, including conformers, cyclic forms, acyclic forms, stereoisomers, tautomers, anomers, and isomers. [Brief description of the drawings]

[0060] [Figure 1]FIG. 1 shows the secretion profiles of IFNγ (left two groups) and IL-2 (right two groups) detected by ELISpot one week after the third administration with saponin and PEK antigen or without PEK antigen.

[0061] [Diagram 2] FIG. 2: Flow cytometry analysis of splenic IFNγ+ (x-axis), CD4+ or CD8+ (y-axis) within the total CD3+ T cell population one week after the third administration of saponin of the invention.

[0062] [Diagram 3] FIG. 2 shows flow cytometry analysis of splenic IL-2+ (x-axis), CD4+ or CD8+ (y-axis) within the total CD3+ T cell population one week after the third administration of saponin of the invention.

[0063] [Figure 4] FIG. 2 shows flow cytometry analysis of splenic TNFα+ (x-axis), CD4+ or CD8+ (y-axis) within the total CD3+ T cell population one week after the third dose of saponin of the invention.

[0064] [Diagram 5] Flow cytometric analysis of T cell populations, representative scatter plots of splenic CD62L+ (x-axis) CD44+ (y-axis) within the total T cell population 1 week after the third dose of sponin of the invention. CD62L low and CD44 high populations were classified as memory T cells and the frequency of viable CD8+ splenocytes expressing IFN-γ, TNF-α or IL-2 is shown. Cytokine positivity was determined when the frequency of positive events exceeded the mean ± SEM of the control group.

[0065] [Figure 6]Figure 1 shows T cell activation 1 week after the third dose of saponins 46-49, 53-56, 56α, 57-62, 64, 66, 77α, 77β, 78, 79, 83, 92, 95. Figure 1 shows the average of spots forming PEK-specific IFNγ+ or IL-2 or TNFα cells in four wells from pooled splenocytes. Cytokine positivity was determined when the frequency of positive events exceeded the mean ± SEM of the control group.

[0066] [Figure 7] Figure 1 shows E7-specific IgG antibody. Serum was collected after each immunization of C57BL / 6 mice immunized with PEK / saponin, and E7 protein-specific IgG antibody in the serum was measured by ELISA. The OD450 value of each serum diluted 10000 times was recorded. Values ​​are expressed as mean ± SEM (n=3).

[0067] [Figure 8] Figure 8(A) shows the median percentage body weight change in mice over 5 days. Values ​​are expressed as mean ± SEM (n=5). Figure 8(B) shows the liver somatic index and Figure 8(C) shows the spleen somatic index. Values ​​are expressed as mean ± SEM (n=5). Mice receiving increasing doses of saponin adjuvant 56 all had a median percentage body weight change of less than 5%. Spleen somatic index and liver somatic index in all experimental groups were unchanged compared to the control group. These data suggest that saponin 56 is a potent and safe candidate as a vaccine adjuvant.

[0068] [Figure 9] FIG. 1 shows the effect of a signal dose of OVA vaccine containing saponin conjugate 56 on E.G7-OVA tumors in female C57BL / 6 mice.

[0069] [Figure 10]FIG. 1 shows survival rates of mice challenged with murine influenza (PR8) after subcutaneous (sc) and intranasal administration of influenza vaccine in combination with compound 56.

[0070] [Figure 11] FIG. 1 shows antibody titers in mice subcutaneously administered SARS-CoV-2 (2 μg or 10 μg) and adjuvants (alum and compound 56). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0071] Saponin conjugates

[0072] The present invention relates to saponin conjugates of formula (I) as defined above, their synthesis, and intermediates thereof.

[0073] The saponin conjugate of formula (I) can be synthesized by the following synthetic steps:

[0074] Step 1-1: Synthesis of trisaccharide donor

[0075] Trisaccharide synthesis was carried out by reacting xylosylimidate 1 and rhamnose acceptor 2 with a catalytic amount of BF 3 ·OEt 2The glycosylation of disaccharide 3 was initiated by glycosylation with NIS to give the disaccharide 3 in 58% yield (Scheme 1). The thiodisaccharide 3 was then subsequently coupled with fucose 4α to provide the desired trisaccharide 5 in 58% yield. Furthermore, these two-step glycosylations can also be carried out in a one-pot manner by first coupling the disaccharide 3 and then sequentially adding fucose 4α and NIS to the reaction mixture to give the trisaccharide 5. The trisaccharide 5 was selectively deprotected using morpholine at room temperature and then acetylated with acetic anhydride. The resulting pentaacetylated trisaccharide 6 was allowed to proceed under hydrogenolysis and imidate formation to provide the trisaccharide donor 10. The optimized trisaccharide approach began with the coupling of xylose 1 with rhamnose 7 to give a quantitative yield of the disaccharide 8, followed by hydrolysis and acetylation to give 9. Glycosylation of disaccharide 9 with fucose 4α gave the trisaccharide 6 in 96% yield. The arabinose-containing trisaccharide 12 and its imidate derivative 13 were also synthesized by glycosylation of the disaccharide 9 and arabinose 11α and the following hydrogenolysis and imidate reactions.

[0076] Scheme 1: shows the synthesis of trisaccharides and their analogues. [ka] (Scheme 1)

[0077] Step 1-2: Synthesis of tetrasaccharide donor

[0078] The synthesis of the tetrasaccharide was achieved by treating glucosyl imidate 15 and rhamnoside 14 with TMSOTf to give disaccharide 16 with the correct β-(1→3) linkage in 51% yield (Scheme 2). After hydrolysis of the thio group and imidate formation, the disaccharide imidate donor 17 was obtained, which was subsequently reacted with fucose 4α to give trisaccharide 18, which was treated with DDQ to remove the PMB function. The resulting trisaccharide acceptor 19 was further coupled with xylosyl donor 1 to achieve tetrasaccharide 20. After confirming the structure by NMR spectroscopy, tetrasaccharide 20 was proceeded under hydrogenolysis and imidate formation to give tetrasaccharide imidates 21 and 22.

[0079] Scheme 2: Synthesis of tetrasaccharides and their analogues. [ka] (Scheme 2)

[0080] Step 2: Synthesis of triterpene building blocks

[0081] The allyl group was first introduced to the C-28 carboxylic acid to give the chelate ester 20 and the echinocyst ester 21 (Scheme 3). To improve the selectivity of the 3-O glycosylation, the 16-OH group of diol 20 was further protected with a triethylsilyl (TES) group by the following three-step synthesis: selective 3-O acetylation, installation of TES at the 16-OH, and then deacetylation to give 60% of the alcohol 22 in three steps.

[0082] Scheme 3: Synthesis of triterpene building blocks and their analogues. [ka] (Scheme 3)

[0083] Step 3: Synthesis of protected bisdesmosidic saponins For the glucuronic acid building block, glucuronic acids bearing benzoyl (Bz), isobutyryl (iPrCO), and pivaloyl (Piv) groups were synthesized (Scheme 4). Reaction of bromide glucoronate 26 with thiotoluene produced thio-glucuronide 27. Compound 27 was then deacetylated and acylated with benzoyl, isobutyryl, and pivaloyl chloride to give 28–30. Following oxidative removal of the thio group and subsequent formation of trichloroacetimidates, glucuronic acid imidates 31–33 were readily available for coupling with chiral acids. Coupling of benzoylated donor 31 with chiral acid ester 23 led to a predominantly orthoester outcome. The increased amount of orthoesters could be due to the flat form of the 2-O-benzoyl group. Therefore, isobutyrylated glucuronic acid 32 was introduced to build a barrier adjacent to the carbonyl position. As a result, the orthoester still predominated at 47%, and the product was isolated at 21%. Nevertheless, this result encouraged us to use the bulkier pivaloyl group. Finally, by using the pivaloyl donor 33, we successfully obtained the conjugated product 34 in 48% yield, and the chiral acid ester 23 was recovered in 29%. The reaction of the echinocyst ester 24 with the benzoylated glucoronate 31 successfully gave the product 36 in 72% yield.

[0084] To deprotect the C-28 carboxylic acid, the benzoyl, pivaloyl and methoxy groups were first hydrolyzed at high temperature under basic conditions. The resulting intermediates were then subjected to benzylation and triethylsilylation to give the fully protected chiral ester 37 and echinocytic ester 38. 1 Based on the H NMR coupling constant analysis, 4 C 1 The form 1 C 4 The original coupling constant between H-1' and H-2' in glucuronide 34 is J H-1′-H-2′ = 7.8 Hz to TES-protected compound 37 J H-1′-H-2′= 4.2 Hz. This reduction in coupling constant was also observed for the other hydrogens on the glucuronide. To further achieve the glucuronide acceptor, the O-allyl ester was hydrolyzed by Pd(OAc)2 catalysis in mildly acidic environment to give compounds 39 and 40. The coupling of oligosaccharide 13 with glucuronide 39 was carried out at -75 °C under the promotion of BF3·OEt2, achieving 41β in excellent yield of 94%. Coupling of arabinose-containing trisaccharide 13 with queratic acid 39 afforded 42β and 42α in 75% and 16%, respectively. Similarly, echinocyst ester 43(β) was also obtained in excellent yield of 96% by glycosylation of imidate 10 with echinocyst acid 40. General coupling conditions for tetrasaccharide donor 21 with queratic acid 39 were applied to give product 44 with anomeric ratio β / α ~ 1 / 1 by TLC analysis. Then, N-phenyltrifluoroacetimidate 22 was utilized, and as a result, saponin 44β was successfully obtained as the major product in 46% yield.

[0085] Scheme 4: Preparation of compounds 41-44. [ka] (Scheme 4)

[0086] Step 4: Synthesis of the amide bond and fully deprotected saponin

[0087] After hydrogenolysis of 41β, sequential amide bond formation was performed using the HBTU / DIPEA coupling system to give a series of conjugated amides. The products then proceeded with acid hydrolysis and methanolysis to provide the desired saponins. Representative saponins in Scheme 5 included aliphatic carbon chains of various lengths, from methyl to octadecyl, and a variety of arylaliphatic, heteroarylaliphatic, and heterocyclic aliphatic compounds.

[0088] Scheme 5: illustrates the preparation of compounds of formula (I) according to an embodiment of the present invention. [ka] (Scheme 5)

[0089] Step 5: Synthesis of saponin analogues

[0090] The synthesis of saponin contained an α-oriented trisaccharide moiety (compound 56α, Scheme 6). Substitution of D-fucose with L-arabinose afforded saponin 77α / β anomers. Echinocyst ester 78 was synthesized, and the b-linked tetrasaccharide ester 79 was also carried out.

[0091] Scheme 6: Shows the preparation of saponin analogs according to embodiments of the present invention. [ka] (Scheme 6)

[0092] Vaccine Compositions

[0093] Another aspect of the present application relates to a vaccine composition comprising an antigen and a saponin analog of the present application as an adjuvant. In some embodiments, the vaccine composition further comprises an additional adjuvant.

[0094] The vaccine compositions of the present application are useful as vaccines to induce active immunity against an antigen in a subject. Any animal that can experience the beneficial effects of the compositions of the present invention is within the scope of subjects that can be treated. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0095] The administration of a vaccine (or the antisera it elicits) may be for either a "prophylactic" or "therapeutic" purpose. Prophylactic administration of a vaccine serves to prevent or reduce subsequent disease symptoms. When provided prophylactically, the vaccine is provided prior to disease symptoms. When provided therapeutically, the vaccine is provided at or after detection of symptoms indicating that an animal may be infected with a pathogen or have a particular cancer. Therapeutic administration of a vaccine serves to reduce actual disease symptoms. Thus, a vaccine can be provided either before the onset of disease proliferation or after the onset of actual proliferation.

[0096] Thus, in one aspect, the invention provides a vaccine comprising one or more bacterial, viral, protozoan, or tumor-associated antigens in combination with one or more compounds of the invention. In some embodiments, the vaccine comprises a single bacterial, protozoan, viral, or tumor-associated antigen in combination with one compound of the invention. In some embodiments, the vaccine comprises two or more bacterial, viral, protozoan, or tumor-associated antigens in combination with a single compound of the invention. In some embodiments, the vaccine comprises a single bacterial, viral, protozoan, or tumor-associated antigen in combination with two or more compounds of the invention.

[0097] In some embodiments, one or more antigens of the provided vaccines are bacterial antigens. In certain embodiments, the bacterial antigen is selected from the group consisting of Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Borrelia burgdorferis, Borrelia spp., Chlamydia trachomatis, Helicobacter pyloris, Chlamydia pneumoniaes, Ureaplasma urealyticums, Mycoplasma pneumoniaes, Staphylococcus spp., Staphylococcus aureus, Streptococcus pyogenes, pyogenes, Streptococcus spp., Streptococcus pneumoniae, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis, Neisseria gonorrhoeae, Bacillus anthracis, Salmonella spp., Salmonella typhi, Vibrio cholera, Pasteurella pestis, Campylobacter spp., Campylobacter jejuni, Clostridium spp. spp.), Clostridium difficile, Corynebacterium diphtheria, Mycobacterium spp.), Mycobacterium tuberculosis, Pseudomonas aeruginosa, Treponema spp., Leptospria spp., Hemophilus ducreyi, Hemophilus influenza, Escherichia coli, Shigella spp., Erlichia spp., Rickettsia spp., and combinations thereof.

[0098] In certain embodiments, one or more antigens of the vaccine provided are virus-associated antigens. In certain embodiments, the virus-associated antigens are antigens associated with a virus selected from the group consisting of influenza virus, parainfluenza virus, mumps virus, adenovirus, respiratory syncytial virus, Epstein-Barr virus, rhinovirus, poliovirus, coxsackievirus, echovirus, measles virus, rubella virus, varicella-zoster virus, herpes virus, herpes simplex virus, parvovirus, cytomegalovirus, hepatitis virus, human papillomavirus, alphavirus, flavivirus, bunyavirus, rabies virus, arenavirus, filovirus, HIV1, HIV2, HTLV-1, HTLV-II, FeLV, bovine LV, FeIV, canine distemper virus, canine infectious hepatitis virus, feline calicivirus, feline rhinotracheitis virus, TGE virus, foot and mouth disease virus, coronavirus, dengue virus, fabivirus, and combinations thereof.

[0099] In certain embodiments, one or more antigens of the vaccines provided are tumor associated antigens. In some embodiments, the tumor associated antigens are selected from the group consisting of killed tumor cells and their lysates, MAGE-1, MAGE-3 and their peptide fragments, human chorionic gonadotropin and its peptide fragments, carcinoembryonic antigen and its peptide fragments, alpha-fetoprotein and its peptide fragments, pancreatic carcinoembryonic antigen and its peptide fragments, prostate specific antigen and its peptide fragments, MUC-1 and its peptide fragments, CA125, CA15-3, CA19-9, CA549, CA195 and its peptide fragments, prostate specific membrane antigen and its peptide fragments, squamous cell carcinoma antigen and its peptide fragments, ovarian cancer antigen and its peptide fragments, pancreatic cancer associated antigen and its peptide fragments, Her1 / neu and its peptide fragments, gp-100 and its peptide fragments, mutant K-ras protein and its peptide fragments, mutant p53 and its peptide fragments, truncated epidermal growth factor receptor, chimeric protein p210 BCR-ABL ,STn,Tn,Lewis x , Lewis y , TF, GM1, GM2, GD2, GD3, Gb3, KH-1, Globo-H, SSEA-4; and mixtures thereof.

[0100] As described above, the provided compounds can be used in cancer vaccines as adjuvants in combination with tumor-associated antigens. In certain embodiments, the vaccines can be used in the treatment or prevention of tumors. In certain embodiments, the tumor is a benign neoplasm. In other embodiments, the tumor is a malignant neoplasm. Any cancer can be treated using the antigen and the compounds of the present invention.

[0101] Another aspect of the present application relates to a method for immunizing a subject with the vaccine composition of the present application.

[0102] formulation

[0103] The saponin analogs of the present application can be combined with a pharma- ceutically acceptable excipient to form a pharmaceutical composition. In certain embodiments, the pharmaceutical composition comprises a pharma- ceutically acceptable amount of the compound of the present invention. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form can vary depending on the host treated and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form is generally that amount of the compound that produces a therapeutic effect. Generally, this amount will range from about 1% to about 99% of the active ingredient, preferably from about 5% to about 70%, and most preferably from about 10% to about 30%.

[0104] Wetting agents, emulsifying agents and lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants may also be present in the composition.

[0105] The formulations of the present invention include those suitable for oral, nasal, topical, rectal, vaginal and / or parenteral administration. The formulations may be presented in convenient unit dosage forms and may be prepared by any method well known in the art of pharmacy. In certain embodiments, the formulations of the present invention comprise an excipient selected from the group consisting of cyclodextrins, liposomes, micelle forming agents (e.g., bile acids), and polymeric carriers (e.g., polyesters and polyanhydrides) and a compound of the present invention. In certain embodiments, the formulations described above render the compound of the present invention orally bioavailable.

[0106] Formulations of the present invention suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavor base, usually sucrose and acacia or tragacanth), powder, granules, or in the form of a solution or suspension in an aqueous or non-aqueous liquid, or in the form of an oil-in-water or water-in-oil liquid emulsion, or in the form of an elixir or syrup, or in the form of a troche (using an inert base, such as gelatin or glycerin, or sucrose and acacia), and / or in the form of a mouthwash, each containing a predetermined amount of a compound of the present invention as an active ingredient. The compounds of the present invention may also be administered as a bolus, electical, or paste.

[0107] The preparations of the present application can be given orally, parenterally, topically, or rectally. Naturally, they are administered in a form suitable for each administration route. For example, they are administered in the form of tablets or capsules, by injection, inhalation, eye lotion, ointment, suppository, etc., by injection, infusion or inhalation, topically by lotion or ointment, rectally by suppository, etc.

[0108] Regardless of the route of administration selected, the saponin analogs of the present application may be used in a suitable hydrated form and / or the pharmaceutical compositions of the present invention may be prepared into a pharma- ceutically acceptable dosage form by conventional methods known to those of skill in the art. Actual dosage levels of the active ingredient in the pharmaceutical compositions of the present invention may be varied to obtain an amount of the active ingredient effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without causing toxicity to the patient.

[0109] The dosage level selected will depend on a variety of factors, including the activity of the particular compound of the invention being used, or an ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound being used, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound being used, or factors well known in the medical arts, such as the age, sex, weight, condition, general health and prior medical history of the patient being treated.

[0110] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required, for example, the physician or veterinarian can start the dosage of the compound of the present invention used in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect, and then gradually increase the dosage until the desired effect is achieved. (Example)

[0111] The following examples are provided to illustrate certain aspects of the present invention and to aid those of skill in the art in practicing the invention, and are not intended to limit the scope of the invention.

[0112] All reagents and solvents were reagent grade and used without further purification unless otherwise noted. Molecular sieves were activated at 200 °C prior to use. Reaction progress was monitored by analytical TLC on 0.25 mm Merck Millipore silica gel 60F254 using p-anisaldehyde, ninhydrin, and ceric ammonium molybdate as visualization agents. Flash column chromatography was performed using 230-400 mesh silica gel.

[0113] Equipment

[0114] NMR spectra were obtained using a Bruker AV-400 (400 MHz) and a Bruker AV-600 (600 MHz). Chemical shifts (δ) are given in ppm and are calculated using CDCl. 3 In the case of 1 H: 7.26 ppm, 13 C: 77.0 ppm; 3 In the case of O.D. 1 H: 3.31 ppm, 13C: 49.0 ppm. Splitting patterns are reported as s (singlet), d (doublet), t (triplet), q (quartet), and m (multiplet). Coupling constants (J) are expressed in Hertz (Hz). Reversed-phase HPLC purification and analysis were performed on a HITACHI D-2000 Elite HPLC system equipped with an autosampler L-2200, UV detector L-2420, and pump L-2130, or a SHIMADZU HPLC system equipped with a system controller CBM-20A, photodiode array detector SPD-M20A, pump LC-20AT, and autosampler SIL-20AHT. Accurate mass measurements were performed on a VG platform electrospray ESI / MS or BioTOFII.

[0115] Synthesis example I [ka] p-Methylphenyl 2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2-O-acetyl-3-O-fluorenylmethyloxycarbonyl-1-thio-α-L-rhamnopyranoside (3).

[0116] 1 (137 mg, 0.33 mmol), 2 (87 mg, 0.16 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in (1.6 mL) 3 ·OEt 2 (approximately 48%, 11 μL, 0.08 mmol) 2 The reaction was completed after 3 hours, and saturated NaHCO 3 The mixture was quenched by the addition of MgSO and allowed to warm to room temperature. 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=1 / 4 to 1 / 2) to give 3 (75 mg, 58%) as a colorless syrup: R f 0.36 (EtOAc / Hexane=1 / 2); 11H NMR (600 MHz, CDCl 3 ) δ 7.78 (d, J = 7.5 Hz, 2H), 7.64 (dd, J = 19.0, 7.4 Hz, 2H), 7.45 - 7.41 (m, 2H), 7.39 - 7.34 (m, 4H), 7.12 (d, J = 8.0 Hz, 2H), 5.54 (dd, J = 3.4, 1.4 Hz, 1H, H-2), 5.33 (d, J = 1.4 Hz, 1H, H-1), 5.21 (t, J = 9.4 Hz, 1H, H-3′), 5.08 (dd, J = 9.6, 3.4 Hz, 1H, H-3), 4.99 (td, J = 9.4, 5.4 Hz, 1H, H-4′), 4.95 (dd, J = 9.4, 7.6 Hz, 1H, H-2′), 4.79 (d, J = 7.6 Hz, 1H, H-1′), 4.57 (dd, J = 13.4, 10.1Hz, 1H, Fmoc C H 2 ), 4.35 - 4.31 (m, 2H, Fmoc C H 2 and C H ), 4.28 - 4.25 (m, 1H, H-5), 4.14 (dd, J = 11.8, 5.4 Hz, 1H, H-5 a ′), 3.84 (t, J = 9.5 Hz, 1H, H-4′), 3.40 (dd, J = 11.8, 9.5 Hz, 1H, H-5 b ′), 2.33 (s, 3H), 2.16 (s, 3H), 2.05 (s, 3H), 2.03 (s, 3H), 1.91 (s, 3H), 1.35 (d, J = 6.2 Hz, 3H, H-6); 13 13C NMR (151 MHz, CDCl 3 ) δ 167.0, 169.9, 169.8, 169.5, 154.0, 143.4, 143.1, 141.3, 141.2, 138.2, 132.6, 129.9, 129.3, 128.0, 127.3, 125.2, 125.0, 120.1, 120.0, 100.8 (C-1′), 85.8 (C-1), 76.1 (C-3), 75.9 (C-4), 72.2 (C-3′), 71.3 (C-2′), 71.2 (C-2), 70.6 (Fmoc C H 2 ), 69.3 (C-4′), 68.1 (C-5), 62.5 (C-5′), 46.6 (Fmoc CH),21.1,20.9,20.7,20.7,20.5,17.5(C-6)ppm; HRMS(ESI-TOF)C 41 H 44 O 14 SNa [M+Na] + The theoretical value was 815.2349 and the measured value was 815.2352. [ka] Benzyl 2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2-O-acetyl-3-O-fluorenylmethyloxycarbonyl-α-L-rhamnopyranosyl-(1→2)-3,4-O-isopropylidene-α-D-fucopyranoside (5).

[0117] 4α (1.05 g, 3.57 mmol), 3 (3.40 g, 4.28 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension of NIS (1.12 g, 4.99 mmol) and BF 3 ·OEt 2 (0.38 mL, 1.42 mmol) 2 The reaction was completed after 1 h, and saturated NaHCO 3 and 10% Na 2 S 2 O 3 The reaction was quenched by the addition of aqueous solution of 1.25 ml of ethyl acetate. The reaction mixture was allowed to warm to room temperature, stirred for 1 h, and filtered. The filtrate was purified by CH 2 Cl 2 Diluted with 10% Na 2 S 2 O 3(aq.) , saturated NaHCO 3 , washed with brine, and MgSO 4 The residue was purified by column chromatography (silica gel; EtOAc / CH 2 Cl 2 The mixture was purified by hexane (hexane = 1 / 1 / 6 to 1 / 1 / 4) to give 5 (2.00 g, 58%) as a white foam: R f 0.44(EtOAc / CH 2 Cl2 / ヘキサン=1 / 1 / 2); 1 H NMR(600 MHz,CDCl 3 )δ 7.78(d,J=7.5 Hz,2H),7.67(d,J=7.4 Hz,1H),7.63(d,J=7.5 Hz,1H),7.42(t,J=7.4 Hz,2H),7.42(t,J=7.4 Hz,2H),7.39-7.27(m,5H),5.50-5.46(m,1H,H-2′),5.19(t,J=9.2 Hz,1H,H-3′′),5.12-5.10(m,2H,H-1′,H-3′),4.99(td,J=9.2,5.5 Hz,1H,H-4′′),4.94(dd,J=9.2,7.6 Hz,1H,H-2′′),4.82(d,J=3.5 Hz,1H,H-1),4.77(d,J=7.5 Hz,1H,H-1′′),4.71(d,J=12.3 Hz,1H,Bn C H 2 ),4.60-4.51(m,2H,Bn C H 2 ,Fmoc C H 2 ),4.37-4.31(m,2H,H-3,Fmoc C H ),4.28(dd,J=10.0,8.1Hz,1H,Fmoc C H 2 ),4.17-4.09(m,2H,H-5,H-5 a ′′),4.02(dd,J=5.3,2.4 Hz,1H,H-4),3.76(dd,J=8.1,3.5 Hz,1H,H-2),3.72(t,J=9.6 Hz,1H,H-4′),3.65-3.60(m,1H,H-5′),3.40(dd,J=11.6,9.7 Hz,1H,H-5 b ′′),2.18(s,3H),2.05(s,3H),2.01(s,3H),1.86(s,3H),1.50(s,3H),1.34(d,J=6.7 Hz,3H,H-6),1.32(s,3H),1.18(d,J=6.2 Hz,3H,H-6′);; HRMS(ESI-TOF)C 50 H 58 O 19 Na [M+Na] +The theoretical value was 985.3463 and the measured value was 985.3476. [ka] Benzyl 2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamnopyranosyl-(1→2)-3,4-O-isopropylidene-α-D-fucopyranoside (6).

[0118] 5 (100 mg, 0.10 mmol) 2 Cl 2 To a stirred suspension in 1000 mL of ethyl acetate (5 mL) was added morpholine (0.5 mL) at room temperature. When the reaction was complete after 1.5 h, the reaction mixture was diluted with CH 2 Cl 2 Dilute with saturated NH 4 Cl, saturated NaHCO 3 , washed with brine, and MgSO 4 The mixture was dried over 1000 ml of ethyl acetate and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane = 1 / 1 to 3 / 2) to give the Fmoc-deprotected product (72 mg, 95%). The Fmoc-deprotected product (462 mg, 0.48 mmol) was dissolved in anhydrous CH 2 Cl 2 To a stirred solution in (10 mL) 2 O (91 μL, 0.96 mmol), Et 3 N (201 μL, 1.4 mmol) and DMAP (6 mg, 0.048 mmol) were added to N 2 The reaction was added at room temperature under atmospheric pressure. After 2 hours, the reaction was complete and the mixture was washed with CH 2 Cl 2 Dilute with H 2 o, washed with brine, and MgSO 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=2 / 3) to give 6 (391 mg, 96%) as a white foam: R f 0.48 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl 3)δ 7.38-7.27(m,5H),5.32(dd,J=3.5,1.6 Hz,1H,H-2′),5.26(dd,J=9.3,3.5 Hz,1H,H-3′),5.13(t,J=9.3 Hz,1H,H-3′′),5.05(d,J=1.5 Hz,1H,H-1′),4.96(td,J=9.3,5.4 Hz,1H,H-4′′),4.91-4.87(dd,J=9.3,7.6 Hz,1H,H-2′′),4.80(d,J=3.6 Hz,1H,H-1),4.71(d,J=12.3 Hz,1H,Bn C H 2 ),4.63(d,J=7.6 Hz,1H,H-1′′),4.54(d,J=12.3 Hz,1H,Bn C H 2 ),4.34(dd,J=8.1,5.4 Hz,1H,H-3),4.14(qd,J=6.7,2.5 Hz,1H,H-5),4.10(dd,J=11.7,5.4 Hz,1H,H-5 a ′′),4.02(dd,J=5.4,2.5 Hz,1H,H-4),3.74(dd,J=8.1,3.6 Hz,1H,H-2),3.62-3.54(m,2H,H-4′,H-5′),3.33(dd,J=11.7,9.5 Hz,1H,H-5 b ′′),2.13(s,3H),2.08(s,3H),2.03(s,3H),2.01(s,6H),1.49(s,3H),1.34(d,J=6.7 Hz,3H,H-6),1.32(s,3H),1.14(d,J=5.6 Hz,3H,H-6′)ppm; HRMS(ESI-TOF)C 37 H 50 O 18 Na [M+Na] + The theoretical value was 805.2889 and the measured value was 805.2898. [ka] Trichloroacetimidoyl 2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamnopyranosyl-(1→2)-3,4-O-isopropylidene-α / β-D-fucopyranoside (10).

[0119] A solution of 6 (1.26 g, 1.6 mmol) and 10% Pd / C (0.2 g) in phosphate buffer (100 mM Na 2 HPO 4 / NaH 2 PO 4 (aq.), pH = 7.0) / THF / MeOH = 1 / 1 / 4 (30 mL) was added to 100 mL of HCl at room temperature. 2 The mixture was stirred under a (balloon) atmosphere. After stirring for 3 days, the mixture was filtered through Celite and concentrated under reduced pressure. The residue was dissolved in CH 2 Cl 2 Dilute with H 2 o, washed with brine, and MgSO 4 The mixture was dried over 500 ml of ethyl acetate, concentrated, and purified by column chromatography (silica gel; EtOAc / hexane = 1 / 1 to 3 / 2) to give the hemiacetal (0.79 g, 71%) as a colorless syrup. 2 Cl 2 To a stirred solution of Cl 3 CCN (16 µL, 0.16 mmol) and DBU (3 µL, 0.021 mmol) were stirred at room temperature with N 2 The mixture was added under atmospheric pressure. After stirring for 1.5 h, the reaction was found to be complete by TLC analysis, and the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; EtOAc / hexane = 1 / 1 to 3 / 2. 0.5% Et 3 N) to give 10 (34 mg, 78%) as a yellow syrup. f 0.46 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl 3)δ 8.67(s,1H),6.30(d,J=3.5 Hz,1H,H-1),5.24(dd,J=3.3,1.8 Hz,1H,H-2′),5.14-5.08(m,2H,H-3′,H-3′′),5.03(d,J=1.8 Hz,1H,H-1′),4.93(td,J=9.4,5.4 Hz,1H,H-4′′),4.86(dd,J=9.4,7.7 Hz,1H,H-2′′),4.58(d,J=7.7 Hz,1H,H-1′′),4.40(dd,J=7.8,5.4 Hz,1H,H-3),4.32(qd,J=6.7,2.5 Hz,1H,H-5),4.12-4.07(m,2H,H-5 a ′′,H-4),3.91(dd,J=7.8,3.5 Hz,1H,H-2),3.81(dq,J=9.6,6.2 Hz,1H,H-5′),3.60(t,J=9.6 Hz,1H,H-4′),3.31(dd,J=11.7,9.7 Hz,1H,H-5 b ′′),2.13(s,3H),2.03(s,3H),2.02(s,3H),2.00(s,3H),1.95(s,3H),1.52(s,3H),1.38(d,J=6.7 Hz,3H,H-6),1.34(s,3H),1.28(d,J=6.2 Hz, 3H, H-6′). [ka] p-Methylphenyl 2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-1-thio-α-L-rhamnopyranoside (9).

[0120] A stirred solution of 8 (4.6 g, 8.1 mmol) in 80% AcOH (100 mL) was heated to 60 °C for 12 h. The resulting mixture was evaporated and azeotroped twice with toluene (50 mL) under reduced pressure. After drying under high vacuum, Ac 2 O (2.2 mL, 23 mmol), Et 3 CH of N (5.2 mL, 38 mmol) and DMAP (9 mg, 0.074 mmol) 2 Cl 2 The solution contains N 2The crude syrup was treated at room temperature under atmospheric pressure. After 2 hours, the reaction was complete and the mixture was washed with CH 2 Cl 2 Dilute with H 2 o, washed with brine, and MgSO 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=2 / 3) to give 9 (3.8 g, 76%) as a white solid: R f 0.19 (EtOAc / Hexane=1 / 2); 1 H NMR (400 MHz, CDCl 3 )δ 7.34(d,J=7.9 Hz,2H),7.11(d,J=7.8 Hz,2H),5.39(brs,1H,H-2),5.26(brs,1H,H-1),5.22(dd,J=9.6,3.1Hz,1H,H-3),5.14(t,J=9.2 Hz,1H,H-3′),5.00-4.93(m,1H,H-4′),4.90(dd,J=9.2,7.6 Hz,1H,H-2′),4.66(d,J=7.6 Hz,1H,H-1′),4.24(dq,J=9.6,6.1Hz,1H,H-5),4.12(dd,J=11.6,5.3 Hz,1H,H-5 a ′),3.72(t,J=9.6 Hz,1H,H-4),3.39-3.30(m,1H,H-5 b ′),2.31(s,3H),2.11(s,3H),2.09(s,3H),2.03(s,3H),2.02(s,6H,Ac×2),1.31(d,J=6.1Hz,3H)ppm;HRMS(ESI-TOF)C 28 H 36 O 13 SNa [M+Na] + The theoretical value was 635.1769 and the measured value was 635.1774. [ka] Benzyl 2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamnopyranosyl-(1→2)-3,4-O-isopropylidene-β-L-arabinopyranoside (12).

[0121] 9 (500 mg, 0.82 mmol), 11α (190 mg, 0.68 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in 1000 mL of 500 sulphuric acid (7 mL), NIS (0.23 g, 1.0 mmol) and TMSOTf (12 µL, 0.066 mmol) were added at -50 °C for 2 h. 2 The reaction was complete after 0.5 h when Et 3 N, saturated NaHCO 3 and 10% Na 2 S 2 O 3 The reaction was quenched by the addition of aqueous solution of 1.25 ml of ethyl acetate. After warming and stirring at room temperature for 1 h, the reaction mixture was filtered and purified by HCl. 2 Cl 2 Dilute with 10% Na 2 S 2 O 3 Aqueous solution, saturated NaHCO 3 , washed with brine, and MgSO 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=2 / 3 to 1 / 1) to give 12 (412 mg, 79%) as a white foam: R f 0.25 (EtOAc / Hexane=2 / 3); 1 H NMR (600 MHz, CDCl 3 )δ 7.38-7.28(m,5H),5.32(dd,J=3.4,1.4 Hz,1H,H-2′),5.26(dd,J=9.2,3.4 Hz,1H,H-3′),5.13(t,J=9.2 Hz,1H,H-3′′),5.05(s,1H,H-1′),4.96(td,J=9.2,5.5 Hz,1H,H-4′′),4.90(dd,J=9.2,7.7 Hz,1H,H-2′′),4.82(d,J=3.4 Hz,1H,H-1),4.73(d,J=12.3 Hz,1H,Bn C H 2 ),4.63(d,J=7.7 Hz,1H,H-1′′),4.53(d,J=12.3 Hz,1H,Bn C H 2),4.36(dd,J=7.9,5.6 Hz,1H,H-3),4.20(d,J=5.6 Hz,1H,H-4),4.10(dd,J=11.7,5.5 Hz,1H,H-5 a ′′),3.98(brs,2H,H-5),3.75(dd,J=7.9,3.4 Hz,1H,H-2),3.61-3.54(m,2H,H-4′,H-5′),3.33(dd,J=11.7,9.6 Hz,1H),2.14(s,3H),2.08(s,3H),2.03(s,3H),2.02-2.01(m,6H,Ac C H 3 ×2),1.50(s,3H),1.32(s,3H),1.14(d,J=5.7 Hz,3H)ppm; HRMS(ESI-TOF)C 36 H 48 O 18 Na [M+Na] + The theoretical value was 791.2733 and the measured value was 791.2735. [ka] Trichloroacetimidoyl 2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamnopyranosyl-(1→2)-3,4-O-isopropylidene-α / β-L-arabinopyranoside (13).

[0122] A solution of 12 (300 mg, 0.39 mmol) and 10% Pd / C (150 mg) in a buffer (100 mM Na 2 HPO 4 / NaH 2 PO 4 (aq.), pH = 7.0) / THF / MeOH = 1 / 1 / 4 (30 mL) was added to 100 mL of HCl at room temperature. 2 The mixture was stirred under a balloon atmosphere. After stirring for 3.5 days, the mixture was filtered through Celite and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 1 to 3 / 2) to give the hemiacetal (166 mg, 63%) as a white foam. The hemiacetal (50 mg, 0.074 mmol) was dissolved in anhydrous CH 2 Cl2 To a stirred solution of Cl 3 CCN (22 µL, 0.22 mmol) and DBU (4.3 µL, 0.029 mmol) were stirred at room temperature with N 2 After stirring for 16 h, the mixture was concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (silica gel; EtOAc / hexane=1 / 1. 0.5% Et 3 N) to give 13 (60 mg, 98%) as a colorless syrup. f 0.43 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl 3 ) δ 8.69(s,1H,N H ),6.30(d,J=3.4 Hz,1H,H-1),5.24(dd,J=3.2,1.3 Hz,1H,H-2′),5.14-5.08(m,2H,H-3′,H-3′′),5.02(d,J=1.3 Hz,1H,H-1′′),4.93(td,J=9.4,5.4 Hz,1H,H-4′′),4.85(dd,J=9.5,7.7 Hz,1H,H-2′′),4.58(d,J=7.7 Hz,1H,H-1′′),4.42(dd,J=7.6,5.7 Hz,1H,H-3),4.31-4.26(m,1H,H-4),4.12(d,J=1.8 Hz,2H,H-5),4.09(dd,J=11.7,5.4 Hz,1H,H-5 a ′′),3.91(dd,J=7.6,3.4 Hz,1H,H-2),3.80(dq,J=9.5,6.2 Hz,1H,H-5′),3.60(t,J=9.5 Hz,1H,H-4′),3.31(dd,J=11.7,9.4 Hz,1H,H-5 b ′′),2.13(s,3H),2.03(s,3H),2.02(s,3H),2.00(s,3H),1.95(s,3H),1.52(s,3H),1.35(s,3H),1.28(d,J=6.2 Hz,3H,H-6′)ppm. [ka] p-Methylphenyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→3)-2-O-acetyl-4-O-(p-methoxybenzyl)-1-thio-α-L-ramopyranoside (16).

[0123] 15 (130 mg, 0.26 mmol), 14 (114 mg, 0.26 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension of 10 µL (0.052 mmol) TMSOTf in 2.5 mL of ethyl acetate was added at -50 °C for 2 h at 4 °C. 2 The reaction was complete after 0.5 h when Et 3 The reaction was quenched by the addition of N, warmed to room temperature, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=1 / 2) to give 16 (102 mg, 51%) as a white foam: R f 0.24 (EtOAc / Hexane=1 / 2); 1 H NMR (600 MHz, CDCl 3)δ 7.29-7.27(m,4H),7.08(d,J=8.0 Hz,2H),6.89(d,J=8.7 Hz,2H),5.39(dd,J=3.4,1.4 Hz,1H,H-2),5.30(d,J=1.4 Hz,1H,H-1),5.18(t,J=9.7 Hz,1H,H-3′),5.06(dd,J=9.7,8.0 Hz,1H,H-2′),5.03(t,J=9.7 Hz,1H,H-4′),4.79(d,J=8.0 Hz,1H,H-1′),4.71(d,J=10.5 Hz,1H),4.46(d,J=10.5 Hz,1H),4.19-4.13(m,2H),4.09(dd,J=12.2,2.2 Hz,1H′),4.06(dd,J=9.5,3.4 Hz,1H),3.79(s,3H,3.70(ddd,J=9.7,5.7,2.2 Hz,1H,H-5′),3.48(t,J=9.5 Hz,1H,H-4),2.29(s,3H),2.10(s,3H),2.09(s,3H),2.01(s,3H),1.98(s,3H),1.83(s,3H),1.25(d,J=6.2 Hz,3H,H-6)ppm; HRMS(ESI-TOF)C 37 H 46 O 15 SNa [M+Na] + The theoretical value was 785.2450 and the measured value was 785.2457. [ka] Trichloroacetimidoyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→3)-2-O-acetyl-4-O-(p-methoxybenzyl)-α / β-L-ramopyranoside (17).

[0124] 16 (37 mg, 0.049 mmol) in acetone / H 2 To a stirred solution in 20O (1 mL) was added NBS (35 mg, 0.19 mmol) at room temperature. After stirring for 2 h, saturated NaHCO 3 and 10% Na 2 S 2 O 3(aq.)The mixture was quenched by the addition of 100 ml of ethyl acetate. The resulting mixture was stirred at room temperature for 1 h and the solvent was removed under reduced pressure. The residue was diluted with CH 2 Cl 2 Dilute with 10% Na 2 S 2 O 3(aq.) , saturated NaHCO 3 , washed with brine, and MgSO 4 The mixture was dried over 1000 ml of ethyl acetate and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=3 / 2) to give the hemiacetal (26 mg, 82%) as a colorless syrup. 2 Cl 2 To a stirred solution of Cl 3 CCN (12 µL, 0.12 mmol) and DBU (2.4 µL, 0.016 mmol) were stirred at room temperature with N 2 After stirring for 1 h, the mixture was concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; EtOAc / hexane=1 / 2. 0.5% Et 3 N) to give 17 (25 mg, 79%) as a white foam. f 0.44 (EtOAc / Hexane=1 / 1); 1 H NMR (400 MHz, CDCl 3 )δ 8.68(s,1H),7.29(d,J=8.4 Hz,2H),6.91(d,J=8.4 Hz,2H),6.15(s,1H,H-1),5.34(s,1H,H-2),5.20(t,J=9.4 C H 2 ), 4.49(d, J=10.3 Hz, 1H, PMBC H 2 ), 4.21-4.13(m,2H,H-3,H-6 a ′), 4.09(t, J=11.0 Hz, 1H, H6 b′),3.89(dq,J=9.6,6.1Hz,1H,H-5),3.81(s,3H,PMB OC H 3 ),3.66(d,J=9.7 Hz,1H,H-5′),3.52(t,J=9.6 Hz,1H,H-4),2.15(s,3H),2.07(s,3H),2.01(s,3H),2.00(s,3H),1.88(s,3H),1.29(d,J=6.1Hz,3H,H-6)ppm. [ka] Benzyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→3)-2-O-acetyl-4-O-(p-methoxybenzyl)-α-L-ramopyranosyl-(1→2)-3,4-O-isopropylidene-α-D-fucopyranoside (18).

[0125] 17 (25 mg, 0.31 mmol), 4α (9 mg, 0.31 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension of 1.5 mL of TMSOTf (1 μL, 0.0062 mmol) was added at -50 °C with N 2 The reaction was completed after 1 h, and Et 3 The reaction was quenched by the addition of N, warmed to room temperature, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexanes=2 / 3) to give 18 (24 mg, 82%) as a white foam: R f 0.43 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl 3)δ 7.34(d,J=7.2 Hz,2H),7.29(t,J=7.5 Hz,2H),7.26-7.24(m,3H),6.89(d,J=8.7 Hz,2H),5.30(dd,J=3.6,1.7 Hz,1H,H-2′),5.16(t,J=9.5 Hz,1H,H-3′′),5.09-5.04(m,H-2′′,H-4′′),5.03(d,J=1.7 Hz,1H,H-1′),4.85(d,J=3.6 Hz,1H,H-1),4.77(d,J=7.9 Hz,1H,H-1′′),4.70(d,J=10.8 Hz,1H,PMB C H 2 ),4.69(d,J=12.2 Hz,1H,Bn C H 2 ),4.51(d,J=12.2 Hz,1H,Bn C H 2 ),4.45(d,J=10.8 Hz,1H,PMB C H 2 ),4.29(dd,J=8.1,5.4 Hz,1H,H-3),4.21(dd,J=12.2,4.9 Hz,1H,H-6 a ′′),4.14-4.09(m,1H,H-5),4.08(dd,J=9.5,3.6 Hz,1H,H-3′),4.04-3.99(m,2H,H-4,H-6 b ′′),3.81(s,3H,PMB OC H 3 ),3.74(dd,J=8.1,3.6 Hz,1H,H-2),3.70-3.61(m,2H,H-5′′,H-5′),3.40(t,J=9.5 Hz,1H,H-4′),2.10(s,3H),2.07(s,3H),2.01(s,3H),1.98(s,3H),1.77(s,3H),1.48(s,3H),1.33(d,J=6.7 Hz,3H,H-6),1.31(s,3H),1.14(d,J=6.2 Hz,3H,H-6′);HRMS(ESI-TOF)C 46 H 60 O 20 Na [M+Na] + The theoretical value is 955.3570 and the measured value is 955.3579.

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[0126] CH of 18 (24 mg, 0.026 mmol) 2 Cl 2 / H 2 To a stirred solution in 1 mL of 18 / O (10 mL) was added DDQ (9 mg, 0.039 mmol) at room temperature. The reaction mixture was stirred for 3 h and saturated NaHCO 3 The resulting mixture was quenched with CH 2 Cl 2 Dilute with saturated NaHCO 3 , washed with brine, and MgSO 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, EtOAc / hexane=1 / 1) to give 19 (17 mg, 81%) as a colorless syrup. f 0.30 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl 3 )δ 7.37-7.29(m,5H),5.24(dd,J=3.6,1.7 Hz,1H,H-2′),5.20(t,J=9.6 Hz,1H,H-3′′),5.04-4.99(m,3H,H-2′′,H-1′,H-4′′),4.87(d,J=3.6 Hz,1H,H-1),4.72(d,J=12.1Hz,1H,Bn C H 2 ),4.70(d,J=7.8 Hz,1H,H-1′′),4.53(d,J=12.1Hz,1H,Bn C H 2 ),4.30(dd,J=8.2,5.3 Hz,1H,H-3),4.22(dd,J=12.3,5.0 Hz,1H,H-6 a ′′),4.12(qd,J=6.7,2.6 Hz,1H,H-5),4.05-4.03(m,2H,H-4,H-6 b′′),3.85(dd,J=9.0,3.6 Hz,1H,H-3′),3.74(dd,J=8.2,3.6 Hz,1H,H-2),3.71(ddd,J=10.1,5.0,2.2 Hz,1H,H-5′′),3.63-3.56(m,2H,H-4′,H-5′),2.46(d,J=2.2 Hz,1H,O H ),2.08(s,3H),2.07(s,3H),2.05(s,3H),2.02(s,3H),2.00(s,3H),1.48(s,3H),1.33(d,J=6.7 Hz,3H,H-6),1.32(s,3H),1.19(d,J=5.7 Hz,3H,H-6′)ppm; HRMS(ESI-TOF)C 38 H 52 O 19 Na [M+Na] + The theoretical value was 835.2995 and the measured value was 835.3000. [ka] Benzyl 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→3)-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4))-2-O-acetyl-α-L-ramopyranosyl-(1→2)-3,4-O-isopropylidene-α-D-fucopyranoside (20).

[0127] 1 (13 mg, 0.031 mmol), 19 (17 mg, 0.021 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in 1 mL of 500 ml of 500 ml of 10 ... TMSOTf (0.4 μL, 0.0021 mmol) was added at -50 °C with N 2 The reaction was complete after 0.5 h when Et 3 The reaction was quenched by the addition of N, warmed to room temperature, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=1 / 1) to give 20 (20 mg, 89%) as a colorless syrup: R f 0.27 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl3 )δ 7.39-7.30(m,5H),5.30(dd,J=3.6,1.8 Hz,1H,H-2′),5.16(t,J=9.5 Hz,1H,H-4′′),5.11(t,J=8.2 Hz,1H,H-3′′′),5.04-4.98(m,2H,H-2′′,H-3′′),4.96(d,J=1.8 Hz,1H,H-1′),4.91(td,J=8.3,4.9 Hz,1H,H-4′′′),4.86(dd,J=8.2,6.3 Hz,1H,H-2′′′),4.82(d,J=3.6 Hz,1H,H-1),4.80(d,J=6.3 Hz,1H,H-1′′′),4.73(d,J=7.7 Hz,1H,H-1′′),4.69(d,J=12.1Hz,1H,Bn C H 2 ),4.50(d,J=12.1Hz,1H,Bn C H 2 ),4.25(dd,J=8.2,5.3 Hz,1H,H-3),4.16-4.09(m,2H,H-6 a ′′,H-5),4.09-4.03(m,2H,H-5 a ′′′,H-3′),4.03-3.99(m,2H,H-4,H-6 b ′′),3.75(t,J=9.5 Hz,1H,H-4′),3.71-3.66(m,2H,H-2,H-5′′),3.54(dq,J=9.5,6.2 Hz,1H,H-5′),3.37(dd,J=11.9,8.3 Hz,1H,H-5 b ′′′),2.19(s,3H),2.08(s,3H),2.08(s,3H),2.08(s,3H),2.04(s,3H),2.01(s,3H),1.99(s,3H),1.97(s,3H),1.46(s,3H),1.33(d,J=6.7 Hz,3H,H-6),1.29(s,3H),1.10(d,J=6.2 Hz,3H,H-6′)ppm; HRMS(ESI-TOF)C 49 H 66 O 26 Na [M+Na] + The theoretical value is 1093.3735 and the measured value is 1093.3734.

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[0128] 20 (255 mg, 0.24 mmol) and 20% Pd(OH) 2 A suspension of 25 mg of 1H2O in 5 mL of THF was heated at room temperature for 1 h. 2 The mixture was stirred under a balloon atmosphere. After stirring for 24 h, the mixture was filtered through Celite and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexanes = 3 / 2 to 2 / 1) to give the hemiacetal (180 mg, 77%) as a white foam. The hemiacetal (20 mg, 0.020 mmol) was dissolved in anhydrous CH 2 Cl 2 To a stirred solution in 1 mL of 500 ml of 500 ml of 1000 ml of N-phenyl-2,2,2-trifluoroacetimidoyl chloride (19 μL, 0.12 mmol) and DBU (3.7 μL, 0.024 mmol) were added at room temperature with N 2 The mixture was stirred for 1.5 h and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel; EtOAc / hexanes = 1 / 1 to 3 / 2. 0.5% Et 3 N) to give 22 (16 mg, 68%) as a colorless syrup. f 0.61 (EtOAc / Hexane=3 / 2); 1 H NMR (600 MHz, CDCl 3)δ 7.33(t,J=7.8 Hz,2H),7.12(t,J=7.4 Hz,1H),6.85(d,J=7.6 Hz,2H),5.58(brs,1H,H-1),5.20(s,1H,H-1′),5.19-5.15(m,2H),5.10(t,J=9.7 Hz,1H),5.06(t,J=7.0 Hz,1H),4.99(dd,J=9.3,7.9 Hz,1H),4.92(d,J=5.1Hz,1H,H-1′′′),4.86(td,J=6.9,4.2 Hz,1H),4.80(dd,J=6.7,5.3 Hz,1H),4.72(d,J=7.9 Hz,1H,H-1′′),4.23-4.08(m,5H),4.00-3.98(m,2H),3.89(brs,1H),3.84-3.77(m,2H),3.66-3.63(m,1H),3.45(dd,J=12.1,7.0 Hz,1H),2.16(s,3H),2.11(s,3H),2.09(s,3H),2.06(s,3H),2.04(s,4H),2.02(s,3H),1.99(s,3H),1.99(s,3H),1.53(s,3H),1.39(d,J=6.3 Hz,3H),1.32(s,3H),1.27(d,J=5.6 Hz,3H)ppm. [ka] 28-O-Allyl-3-O-(methyl 2,3,4-tri-O-pivaloyl-β-D-glucopyranosyluronate) chelate (34).

[0129] 33 (500 mg, 0.83 mmol), 23 (435 mg, 0.83 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension of B(PhF 5 ) 3 (42 mg, 0.083 mmol) at room temperature with N 2 The reaction was complete after 0.5 h when Et 3 The reaction was quenched by the addition of N. It was filtered and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / CH 2 Cl2 The mixture was purified by hexane (hexane = 1 / 1 / 6) to give 34 (385 mg, 48%) as a white solid: R f 0.57 (EtOAc / Hexane = 1:2); 1 H NMR (600 MHz, CDCl 3 )δ 9.40(s,1H,H-23),5.88-5.81(m,1H,total internal alkenyl C H ),5.37(t,J=3.3 Hz,1H,H-12),5.31-5.27(m,2H,H-3′,All terminal alkenyl C H a ), 5.20-5.16 (m, 2H, H-4′, all terminal alkenyl C H b ), 4.98(t,J=8.0 Hz,H-2′), 4.52-4.44(m,4H,H-1′,H-16,allylic C H 2 ),3.99(d,J=10.0 Hz,1H,H-5′),3.84(dd,J=11.8,4.7 Hz,1H,H-3),3.72(s,3H,OC H 3 ,3.05(dd,J=14.3,4.1Hz,1H,H-18),2.15(t,J=13.6 Hz,1H,H-19 a ),1.92-1.85(m,4H),1.80-1.70(m,4H),1.68-1.63(m,3H),,1.50-1.41(m,2H),1.35-1.28(m,4H),1.25-1.17(m,3H),1.14 (s,9H),1.12-1.10(m,10H),1.09-1.08(m,12H),1.05-0.97(m,2H),0.96(s,3H),0.94(s,3H),0.89(s,3H),0.70(s,3H)ppm; HRMS(ESI-TOF)C 55 H 84 O 14 Na [M+Na] + The theoretical value was 991.5753 and the measured value was 991.5758. [ka] 28-O-Allyl-3-O-(benzyl 2,3,4-tri-O-triethylsilyl-β-D-glucopyranosyluronate)-16-O-triethylsilyl chelate (37).

[0130] To a stirred solution of 34 (358 mg, 0.37 mmol) in THF (15 mL) was added 1.0 N KOH (3 mL) and heated at reflux at 66° C. The reaction mixture was stirred for 24 h and cooled to room temperature. The reaction mixture was purified by Amberlyst IR-120H + The residue was neutralized with 1,000 cc of 1 ... 2 CO 3 (102 mg, 0.73 mmol) at room temperature. The reaction mixture was stirred for 2 h and then diluted with CH 2 Cl 2 Dilute with H 2 o, washed with brine, and MgSO 4 The residue was purified by column chromatography (silica gel, MeOH / CH 2 Cl 2 =1 / 15) to give the benzyl ester as a yellow solid. 2 Cl 2 To a stirred solution in 5 mL of N 2 The reaction mixture was stirred for 2 h and saturated NaHCO 3 The mixture was quenched by the addition of CH 2 Cl 2 Dilute with saturated NaHCO 3 , washed with brine, and MgSO 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, EtOAc / hexane=1 / 40) to give 37 (310 mg, 67%, 3 steps) as a colorless syrup: R f 0.71 (EtOAc / Hexane=1 / 10); 1 H NMR (400 MHz, CDCl 3)δ 9.34(s,1H,H-23),7.39-7.30(m,5H),5.90-5.80(m,1H,Total internal alkenyl C H ), 5.35(s,1H,H-12), 5.29(d,J=17.2 Hz,1H,All terminal alkenyl C H a ), 5.20(d, J = 10.7 Hz, 1H, all terminal alkenyl C H b ), 5.17-5.15(m,2H,Bn C H 2 ), 4.58-4.57 (m, 2H, H-1′, H-16), 4.53-4.40 (m, 2H, allyl ic C H 2 ),4.27(dd,J=6.2,4.9 Hz,1H,H-4′),4.22(d,J=6.2 Hz,1H,H-5′),3.81(dd,J=11.4,3.8 Hz,1H,H-3),3.61(d,J=4.9 Hz,1H,H-3′),3.57(d,J=3.4 Hz,1H,H-2′),3.02(d,J=11.6 Hz,1H,H-18),2.23(t,J=13.5 Hz,1H,H-19),1.99(d,J=10.8 Hz,1H),1.91-1.79(m,4H),1.75-1.53(m,5H),1.48-1.31(m,5H),1.30-1.22(m,1H) ,1.20-1.23(m,3H),1.11-1.06(m,4H),1.03-0.85(m,47H),0.72-0.52(m,27H)ppm; HRMS(ESI-TOF)C 70 H 120 O 11 S 4 Na [M+Na] + The theoretical value was 1271.7800 and the measured value was 1271.7829. [ka] 3-O-(benzyl 2,3,4-tri-O-triethylsilyl-β-D-glucopyranosyluronate)-16-O-triethylsilyl chiral acid (39).

[0131] 37 (0.60 g, 0.48 mmol) and PPh3 To a stirred solution of (0.31 g, 1.2 mmol) in 1,4-dioxane (5 mL) was added a solution of formic acid (0.38 mL, 10 mmol) in 1,4-dioxane (2.5 mL) premixed with Et 3 Pd(OAc) in N (1.3 mL, 9.6 mmol) and 1,4-dioxane (2.5 mL) 2 (54 mg, 0.24 mmol) was added at room temperature. The reaction mixture was stirred for 12 h and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, EtOAc / hexane=1 / 6) to give 39 (0.48 g, 83%) as a white foam: R f 0.25 (EtOAc / Hexane=1 / 6); 1 H NMR (600 MHz, CDCl 3 )δ 9.36(s,1H,H-23),7.37-7.30(m,5H),5.34(t,J=3.4 Hz,1H,H-12),5.16(s,2H,Bn C H 2 ),4.59(d,J=4.4 Hz,1H,H-1′),4.53(s,1H,H-16),4.28(dd,J=6.2,5.3 Hz,1H,H-4′),4.22(d,J=6.2 Hz,1H,H-5′),3.82(dd,J=11.8,4.6 Hz,1H,H-3),3.61(d,J=5.3 Hz,1H,H-3′),3.58(d,J=4.4,1H,H-2′),2.95(dd,J=14.3,4.0 Hz,1H,H-18),2.21(t,J=13.6 Hz,1H,H-19 a ),2.03-1.95(m,1H),1.90-1.80(m,4H),1.78-1.61(m,4H),1.57(d,J=13.4 Hz,1H),1.47-1.37(m,2H),1.35(s,3H),1.27(d,J=13.6 HRMS(ESI-TOF)C 67 H 116 O 11 S 4 Na [M+Na] +The theoretical value was 1231.7487 and the measured value was 1231.7507. [ka] 28-O-Allyl-3-O-(methyl 2,3,4-tri-O-benzoyl-β-D-glucopyranosyluronate) echinocyst ester (36).

[0132] 31 (20 mg, 0.030 mmol), 24 (7.7 mg, 0.015 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 (0.6 mL) of B(PhF 5 ) 3 (1.5 mg, 0.0030 mmol) at room temperature with N 2 The reaction was complete after 0.5 h when Et 3 The reaction was quenched by the addition of N, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / CH 2 Cl 2 The mixture was purified by hexane (hexane = 1 / 1 / 6) to give 36 (11 mg, 72%) as a white solid: R f 0.42 (EtOAc / Hexane = 1:2); 1 H NMR (600 MHz, CDCl 3 )δ 7.95-7.91(m,4H),7.85(dd,J=8.4,1.3 Hz,2H),7.53-7.49(m,2H),7.45-7.42(m,1H),7.40-7.35(m,4H),7.31-7.28(m,2H),5.91(t,J=9.7 Hz,1H,H-3′),5.89-5,82(m,1H,Total internal alkenyl C H ),5.64(t,J=9.7 Hz,1H,H-4′),5.59(dd,J=9.7,7.8 Hz,1H,H-2′),5.39(t,J=3.6 Hz,1H,H-12),5.29(dt,J=17.2,1.5 Hz,1H,All terminal alkenyl C H a ), 5.20(dt, J = 10.5, 1.3 Hz, 1H, all terminal alkenyl C Hb ), 4.89 (d, J = 7.8 Hz, 1H, H-1′), 4.53 - 4.45 (m, 3H, H-16, allylic C H 2 ), 4.30 (d, J = 9.7 Hz, 1H, H-5′), 3.69 (s, 3H, OC H 3 ), 3.16 (dd, J = 11.7, 4.6 Hz, 1H, H-3), 3.06 (dd, J = 14.4, 4.3 Hz, 1H, H-18), 2.14 (dd, J = 13.7 Hz, 1H, H-19 a ), 1.90 - 1.85 (m, 4H, H-2 a , H-11 ab , H-22 a ), 1.82 - 1.71 (m, 4H, H-2 b , H-15 a , H-21 a , H-22 b ), 1.65 - 1.61 (m, 1H, H-1 a ), 1.55 - 1.52 (m, 2H, H-9, 16 - O H ), 1.44 - 1.34 (m, 2H, H-6 a , H-7 a ), 1.34 - 1.30 (m, 4H, H-15 b , H-27), 1.26 - 1.19 (m, 3H, H-6 b , H-7 b , H-21 b ), 1.14 - 1.10 (m, 1H, H-19 b ), 0.97 (s, 3H, H-30), 0.94 - 0.92 (m, 1H, H-1 b ), 0.90 (s, 3H, H-29), 0.88 (s, 3H, H-25), 0.71 (s, 3H, H-23), 0.70 - 0.65 (m, 4H, H-5, H-26), 0.62 (s, 3H, H-24) ppm; HRMS(ESI - TOF) C 61 H 74 O 13 Na [M + Na] + The theoretical value at [M+Na] was 1037.5022, and the measured value was 1037.5026.

Chemical formula

[0133] To a stirred solution of 36 (0.45 g, 0.41 mmol) in THF (25 mL) was added 1.0 N KOH (5 mL) and heated at 45° C. The reaction mixture was stirred for 12 h and cooled to room temperature. The reaction mixture was purified by Amberlyst IR-120H + The residue was neutralized with 1,000 cc of 1 ... 2 CO 3 (113 mg, 0.82 mmol) at room temperature. After stirring for 12 h, the reaction mixture was diluted with CH 2 Cl 2 Dilute with H 2 o, washed with brine, and MgSO 4 The residue was purified by column chromatography (silica gel, MeOH / CH 2 Cl 2 = 1 / 20) to obtain the benzyl ester. 2 Cl 2 To a stirred solution of TESOTf (0.74 mL, 3.3 mmol) and 2,6-lutidine (0.48 mL, 4.1 mmol) in 1000 mL of N 2 The reaction mixture was stirred for 2 h and saturated NaHCO 3 The mixture was quenched by the addition of CH 2 Cl 2 Dilute with saturated NaHCO 3 , washed with brine, and MgSO 4 The mixture was dried at 40° C. and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, EtOAc / hexane=1 / 40) to give 38 (0.21 g, 41%) as a colorless syrup: R f 0.41 (EtOAc / Hexane=1 / 20); 1 H NMR (400 MHz, CDCl 3) δ 7.39-7.28 (m, 5H), 5.92-5.82 (m, 1H, all internal alkenyl C H ), 5.35(s,1H,H-12), 5.33-5.27(m,1H,all terminal alkenyl C H ), 5.20(d, J = 10.3 Hz, 1H, all terminal alkenyl C H ),5.16(s,2H,Bn C H 2 ),4.77(d,J=4.2 Hz,1H,H-1′),4.59(s,1H,H-16),4.53-4.43(m,2H All C H 2 ),4.34(dd,J=6.1,5.1Hz,1H,H-4′),4.27(d,J=6.1Hz,1H,H-5′),3.73(d,J=4.2 Hz,1H,H-2′),3.65(d,J=5.1Hz,1H,H-3′),3.06-2.98(m,2H,H-3,H-18),2.23(t,J=13.5 Hz,1H,H-19 a ),1.96(d,J=10.8 Hz,1H),1.89-1.80(m,4H),1.77-1.62(m,3H),1.58-1.40(m,4H),1.34(s,3H),1.31-1.23(m,3H) ,1.16-1.04(m,2H),1.04-0.90(m,42H),0.89-0.84(s,7H),0.78(s,3H),0.72-0.55(m,28H)ppm; HRMS(ESI-TOF)C 70 H 122 O 10 S 4 Na [M+Na] + The theoretical value was 1257.8007 and the measured value was 1257.8019. [ka] 3-O-(benzyl 2,3,4-tri-O-triethylsilyl-β-D-glucopyranosyluronate)-16-O-triethylsilylechinocystic acid (40).

[0134] 38 (194 mg, 0.16 mmol) and PPh 3To a stirred solution of (1.03 mg, 0.39 mmol) in 1,4-dioxane (4 mL) was added a solution of premixed formic acid (150 μL, 3.3 mmol) / Et 3 Pd(OAc) in N (430 μL, 3.1 mmol) and 1,4-dioxane (2 mL) 2 (17 mg, 0.078 mmol) was added at room temperature. The reaction mixture was stirred for 12 h and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, EtOAc / hexane=1 / 6) to give 40 (168 mg, 89%) as a white foam: R f 0.58 (EtOAc / Hexane=1 / 6); 1 H NMR (400 MHz, CDCl 3 )δ 7.40-7.28(m,5H),5.34(s,1H,H-12),5.16(s,2H,Bn C H 2 ),4.78(d,J=4.3 Hz,1H,H-1′),4.55(s,1H,H-16),4.34(t,J=5.5 Hz,1H,H-4′),4.27(d,J=5.8 Hz,1H,H-5′),3.73(d,J=3.9 Hz,1H,H-2′),3.66(d,J=5.0 Hz,1H,H-3′),3.02(dd,J=11.5,3.7 Hz,1H,H-3),2.95(d,J=11.3 Hz,1H,H-18),2.21(t,J=13.5 Hz,1H,H-19 a ),1.96(d,J=11.1Hz,1H),1.92-1.62(m,7H),1.62-1.40(m,4H),1.34(s,3H),1.31-1.23(m,3H) ,1.17-1.04(m,2H),1.02-0.91(m,42H),0.90-0.84(s,7H),0.79(s,3H),0.73-0.55(m,28H)ppm; HRMS(ESI-TOF)C 67 H 118 O 10 S 4 Na [M+Na] + The theoretical value was 1217.7694 and the measured value was 1217.7703. [ka] 3-O-(benzyl 2,3,4-tri-O-triethylsilyl-β-D-glucopyranosyluronate)-28-O-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamnopyranosyl-(1→2)-3,4-O-isopropylidene-β / α-D-fucopyranosyl)-16-O-triethylsilyl chiral acid ester (41β / α)

[0135] 10 (276 mg, 0.33 mmol), 39 (266 mg, 0.22 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in (11 mL) 3 ·OEt 2 (approximately 48%, 12 μL, 0.044 mmol) at -75 °C with N 2 The reaction was complete after 0.5 h when Et 3 The reaction was quenched by the addition of N, warmed to room temperature, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / CH 2 Cl 2 The mixture was purified by hexane (hexane = 2 / 1 / 5) to give 41β (391 mg, 94%) as a colorless syrup. f 0.64 (EtOAc / Hexane = 1:1); 1 H NMR (600 MHz, CDCl 3 )δ 9.35(s,1H,H-23),7.40-7.28(m,5H),5.41(d,J=7.5 Hz,1H,H-1′),5.34(t,J=3.5 Hz,1H,H-12),5.25(dd,J=3.5,1.6 Hz,1H,H-2′′),5.21(dd,J=9.8,3.5 Hz,1H,H-3′′),5.16(s,2H,Bn C H 2),5.13(t,J=9.2 Hz,1H,H-3′′′),4.98(d,J=1.6 Hz,1H,H-1′′),4.96(td,J=9.2,5.5 Hz,1H,H-4′′′),4.85(dd,J=9.2,7.7 Hz,1H,H-2′′′),4.63(d,J=7.7 Hz,1H,H-1′′′),4.58(d,J=4.3 Hz,1H,H-1′′′′),4.49(s,1H,H-16),4.27(t,J=6.3 Hz,1H,H-4′′′′),4.22(d,J=6.3 Hz,1H,H-5′′′′),4.18(t,J=6.0 Hz,1H,H-3′),4.13(dd,J=11.7,5.4 Hz,1H,H-5 a ′′′),4.01(dd,J=6.0,2.0 Hz,1H,H-4′),3.87(qd,J=6.5,2.0 Hz,1H,H-5′),3.84-3.79(m,2H,H-3,H-5′′),3.66(dd,J=7.5,6.3 Hz,1H,H-2′),3.64-3.60(m,2H,H-3′′′′,H-4′′),3.57(dd,J=4.3,0.9 Hz,1H,H-2′′′′),3.34(dd,J=11.7,9.3 Hz,1H,H-5 b ′′′),2.94(dd,J=14.2,4.0 Hz,1H,H-18),2.23(t,J=13.6 Hz,1H,H-19 a ),2.13(s,3H),2.06(s,3H),2.03(s,3H),2.00(s,3H,),1.99-1.97(m,4H,H-2,Ac C H 3 ),1.88-1.75(m,5H,H-11 ab ,H-21 a ,H-22 ab ),1.73-1.65(m,2H,H-2 a ,H-15 a ),1.63-1.60(m,1H,H-9),1.57-1.55(m,1H,H-1 a ),1.53(s,3H,イソプロピリデン C H 3 ),1.47-1.40(m,2H,H-6 a ,H-7 a), 1.35(s,3H,H-27), 1.34(s,3H,isopropylidene C H 3 ),1.29(d,J=6.5 Hz,4H,H-6′),1.27-1.24(m,H-6′′,H-15 b ), 1.19-1.16(m,2H,H-5,H-6 b ), 1.12(d, J=7.8 Hz, 1H, H-21 b ),1.09(s,3H,H-24),1.05(dd,J=12.1,4.0 Hz,1H,H-19 b ), 1.01-0.89(m,44H,TES C H 3 ,H-1 b ,H-7 b ,H-25,H-30),0.89(s,3H,H-29),0.73(s,3H,H-26),0.69-0.55(m,24H,TES C H 2 )ppm; HRMS(ESI-TOF)C 97 H 158 O 28 S 4 Na [M+Na] + The theoretical value was 1906.9937 and the measured value was 1906.9959. 41α:R f 0.69 (EtOAc / Hexane=1:1), 1 H NMR (600 MHz, CDCl 3 )δ 9.34(s,1H,H-23),7.37-7.30(m,5H),6.02(d,J=3.7 Hz,1H,H-1′′),5.39(t,J=3.6 Hz,1H,H-12),5.21(dd,J=3.5,1.7 Hz,1H,H-2′′′),5.16(s,2H,Bn C- H 2),5.11(t,J=9.0 Hz,1H,H-3′′′′),5.08-5.06(m,2H,H-1′′′,H-3′′′),4.93(td,J=9.0,5.3 Hz,1H,H-4′′′′),4.80(dd,J=9.0,7.4 Hz,1H,H-2′′′′),4.66(d,J=7.4 Hz,1H,H-1′′′′),4.58(d,J=4.4 Hz,1H,H-1′),4.56(s,1H,H-16),4.27(t,J=6.3 Hz,1H,H-4′),4.21(d,J=6.3 Hz,1H,H-5′),4.19(dd,J=8.0,5.1Hz,1H,H-3′′),4.13-4.09(dd,J=11.8,5.2 Hz,1H,H-5 a ′′′′),4.06-4.01(m,2H,H-4′′,H-5′′),3.87(dd,J=8.0,3.7 Hz,1H,H-2′′),3.81(dd,J=11.8,4.7 Hz,1H,H-3),3.70-3.64(m,1H,H-5′′′),3.63 - 3.58(m,2H,H-4′′′,H-3′),3.57(dd,J=4.4,1.1Hz,1H,H-2′),3.35(dd,J=11.8,9.0 Hz,1H,H-5 b ′′′′),3.00(dd,J=14.3,4.1Hz,1H,H-18),2.19(t,J=13.2 Hz,1H,H-19 a ),2.13(s,3H),2.03(s,3H),2.02(s,3H),2.02(s,3H),2.00-1.97(m,1H,H-2 a ),1.96(s,3H),1.95-1.83(m,3H,H-22 a ,H-11 ab ),1.81-1.65(m,3H,H-21 a ,H-22 b ,H-2 b ),1.62-1.52(m,3H,H-9,H-1 a ,H-15 a ),1.51(s,3H,イソプロピリデン C H 3 ),1.48-1.40(m,2H,H-6 a ,H-7 a), 1.35(s,3H,H-27), 1.34(s,3H,isopropylidene C H 3 ),1.32(d,J=6.5 Hz,3H,H-6′′),1.29-1.27(m,4H,H-15 b ,H-6′′′),1.23-1.11(m,3H,H-21 b ,H-5,H-6 b ), 1.09-1.05(m,4H,H-24,H-19 b ), 1.01(m,TES C H 3 ×3), 0.96-0.90(m, 35H, H-1 b ,H-7 b ,H-25,H-30,TES C H 3 ×9),0.88(s,3H,H-29),0.73(s,3H,H-26),0.67(m,6H,TES C H 2 ×3), 0.62-0.54(m, 18H, TES C H 2 ×9)ppm; HRMS(ESI-TOF)C 97 H 158 O 28 S 4 Na [M+Na] + The theoretical value was 1906.9937 and the measured value was 1906.9967. [ka] 3-O-(benzyl 2,3,4-tri-O-triethylsilyl-β-D-glucopyranosyluronate)-28-O-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamnopyranosyl-(1→2)-3,4-O-isopropylidene-β / α-L-arabinopyranosyl)-16-O-triethylsilyl chiral acid ester (42β / α).

[0136] 13 (67 mg, 0.081 mmol), 39 (65 mg, 0.054 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2To a stirred suspension in (6 mL) 3 ·OEt 2 (approximately 48%, 1.5 μL, 0.0058 mmol) at -75 °C with N 2 The reaction was complete after 0.5 h when Et 3 The reaction was quenched with N and allowed to warm to room temperature. The resulting mixture was diluted with CH 2 Cl 2 Dilute with saturated NaHCO 3 , washed with brine, and MgSO 4 The residue was purified by column chromatography (silica gel; EtOAc / CH 2 Cl 2 / hexane = 2 / 1 / 5) to give 101 (101β: 76 mg, 75%, 101α: 15 mg, 16%) as a colorless syrup. f 0.24(EtOAc / CH 2 Cl 2 / Hexane=2 / 1 / 5); 1 H NMR (600 MHz, CDCl 3 )δ 9.36(s,1H,H-23),7.39-7.29(m,5H),5.76(d,J=3.8 Hz,1H,H-1′),5.35(s,1H,H-12),5.21(s,1H,H-2′′),5.19-5.15(m,3H,H-3′′,Bn C H 2 ),5.13(t,J=9.2 Hz,1H,H-3′′′),4.97-4.92(m,2H,H-1′′,H-4′′′),4.86(dd,J=9.2,7.6 Hz,1H,H-2′′′),4.63(d,J=7.6 Hz,1H,H-1′′′),4.58(d,J=4.2 Hz,1H,H-1′′′′),4.55(s,1H,H-16),4.35(q,J=6.2 Hz,1H,H-4′),4.27(t,J=5.8 Hz,1H,H-4′′′′),4.21(d,J=6.3 Hz,1H,H-5′′′′),4.17(t,J=5.4 Hz,1H,H-3′),4.12(dd,J=11.5,5.1Hz,1H,H-5 a ′′′),3.85-3.76(m,4H,H-3,H-2′,H-5 a′,H-5′′),3.71(dd,J=11.7,8.2 Hz,1H,H-5 b ′),3.64-3.60(m,2H,H-4′′,H-3′′′′),3.57(d,J=4.2 Hz,1H,H-2′′′′),3.33(t,J=10.6 Hz,1H,H-5 b ′′′),2.98(d,J=11.8 Hz,1H,H-18),2.24(t,J=13.5 Hz,1H,H-19 a ),2.13(s,3H),2.07(s,3H),2.03(s,3H),2.01(s,3H),2.00-1.97(m,4H,Ac C H 3 ,H-2 a ),1.89-1.65(m,7H,H-2 b ,H-11 ab ,H-15 a ,H-21 a ,H-22 ab ),1.63-1.60(m,1H,H-9),1.56(d,J=13.5 Hz,1H,H-1 a ),1.51(s,3H,C H 3 )1.48-1.40(m,2H,H-6 a ,H-7 a ),1.35(s,6H,H-27,C H 3 ),1.27-1.24(m,4H,H-15 b ,H-6′′),1.20-1.12(m,3H,H-5,H-6 b ,H-21 b ),1.09(s,3H,H-24),1.06(d,J=13.2 Hz,1H,H-19 b ),1.01-0.90(m,44H,H-1 b ,H-7 b ,H-25,H-30,TES C H 3 ),0.89(s,3H,H-29),0.72(s,3H,H-26),0.69-0.53(m,24H)ppm; HRMS(ESI-TOF)C 96 H 156 O 28 Si 4 Na [M+Na]+ The theoretical value was 1892.9793 and the measured value was 1892.9791. 42α:R f 0.29 (EtOAc / CH 2 Cl 2 / hexane = 2 / 1 / 5); 1 1H NMR (600 MHz, CDCl 3 ) δ 9.35 (s, 1H, H-23), 7.39 - 7.30 (m, 5H), 6.07 (d, J = 3.3 Hz, 1H, H-1′′), 5.39 (t, J = 3.5 Hz, 1H, H-12), 5.22 (dd, J = 3.5, 1.3 Hz, 1H, H-2′′′), 5.16 (s, 2H, Bn C H 2 ), 5.11 (t, J = 8.9 Hz, 1H, H-3′′′′), 5.07 (dd, J = 9.7, 3.5 Hz, 1H, H-3′′′), 5.05 (d, J = 1.3 Hz, 1H, H-1′′′), 4.94 (td, J = 8.9, 5.3 Hz, 1H, H-4′′′′), 4.81 (dd, J = 8.9, 7.4 Hz, 1H, H-2′′′′), 4.67 (d, J = 7.4 Hz, 1H, H-1′′′′), 4.58 (d, J = 4.4 Hz, 1H, H-1′), 4.57 (s, 1H, H-16), 4.27 (t, J = 5.8 Hz, 1H, H-4′), 4.25 - 4.20 (m, 3H, H-3′′, H-4′′, H-5′), 4.11 (dd, J = 11.8, 5.3 Hz, 1H, H-5 a ′′′′), 4.05 - 4.01 (d, J = 13.4 Hz, 1H, H-5 a ′′), 3.91 - 3.84 (m, 2H, H-2′′, H-5 b ′′), 3.81 (dd, J = 11.8, 4.6 Hz, 1H, H-3), 3.69 (dq, J = 9.5, 6.1Hz, 1H, H-5′′′), 3.64 - 3.59 (m, 2H, H-4′′′, H-3′), 3.57 (d, J = 4.4 Hz, 1H, H-2′), 3.36 (dd, J = 11.8, 9.1Hz, 1H, H-5 b ′′′′), 2.99 (dd, J = 14.4, 4.0 Hz, 1H, H-18), 2.20 (t, J = 13.6 Hz, 1H, H-19 a),2.14(s,3H),2.04(s,3H),2.03(s,3H),2.02(s,3H),2.00-1.99(m,1H,H-2 a ),1.97(s,3H),1.95-1.83(m,3H,H-22 a ,H-11 ab ), 1.81-1.65(m,3H,H-21 a ,H-22 b ,H2 b ),1.65-1.62(m,1H,H-9),1.58-1.53(m,2H,H-1 a ,H-15 a ), 1.51(s,3H,isopropylidene C H 3 ), 1.45-1.40(m,2H,H-6 a ,H-7 a ), 1.36-1.35(m,6H,H-27,isopropylidene C H 3 ),1.32(dd,J=14.5,1.6 Hz,1H,H-15 b ),1.29(d,J=6.1Hz,3H,H-6′′′),1.23-1.15(m,3H,H-5,H-6 b ,H-21 b ), 1.10-1.05(m,4H,H-19 b ,H-24),1.04-0.89(m,44H,H-1 b ,H-7 b ,H-25,H-30,TES C H 3 ×12),0.88(s,3H,H-29),0.73(s,3H,H-26),0.71-0.54(m,24H,TES C H 2 ×12)ppm; HRMS(ESI-TOF)C 96 H 156 O 28 S 4 Na [M+Na] + The theoretical value was 1892.9793 and the measured value was 1892.9780. [ka] 3-O-(benzyl 2,3,4-tri-O-triethylsilyl-β-D-glucopyranosyluronate)-28-O-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamnopyranosyl-(1→2)-3,4-O-isopropylidene-β-D-fucopyranosyl)-16-O-triethylsilyl echinocyst ester (43β).

[0137] 10 (166 mg, 0.20 mmol), 40 (158 mg, 0.13 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 (6.5 mL) 3 ·OEt 2 (approximately 48%, 7.0 μL, 0.026 mmol) at -75 °C with N 2 The reaction was complete after 0.5 h when Et 3 The reaction was quenched with N and allowed to warm to room temperature. The resulting mixture was diluted with CH 2 Cl 2 Dilute with saturated NaHCO 3 , washed with brine, and MgSO 4 The residue was purified by column chromatography (silica gel; EtOAc / CH 2 Cl 2 / hexane = 2 / 1 / 5) to give 43β (237 mg, 96%) as a colorless syrup. f 0.23(EtOAc / CH 2 Cl 2 / Hexane=2 / 1 / 5); 1 H NMR (600 MHz, CDCl 3 )δ 7.37-7.29(m,5H),5.42(d,J=7.5 Hz,1H,H-1′),5.34(s,1H,H-12),5.26(dd,J=3.4,1.5 Hz,1H,H-2′′),5.21(dd,J=9.8,3.4 Hz,1H,H-3′′),5.16(d,J=3.2 Hz,2H,Bn C H 2),5.13(t,J=9.3 Hz,1H,H-3′′′),4.99(d,J=1.5 Hz,1H,H-1′′),4.95(td,J=9.3,5.4 Hz,1H,H-4′′′),4.85(dd,J =9.3,7.6 Hz,1H,H-2′′′),4.77(d,J=4.4 Hz,1H,H-1′′′′),4.64(d,J=7.6 Hz,1H,H-1′′′),4.50(s,1H,H-16),4.33(dd,J=6.1,5.8 Hz,1H,H-4′′′′),4.26(d,J=6.1Hz,1H,H-5′′′′),4.18(t,J=6.0 Hz,1H,H-3′),4.12(dd,J=11.7,5.4 Hz,1H,H-5 a ′′′),4.01(dd,J=6.0,1.8 Hz,1H,H-4′),3.89-3.81(m,2H,H-5′,H-5′′),3.72(d,J=4.4 Hz,1H,H-2′′′′),3.68-3.61(m,3H,H-2′,H-3′′′′,H-4′′),3.34(dd,J=11.9,9.3 Hz,1H,H-5 b ′′′),3.01(dd,J=11.7,4.3 Hz,1H,H-3),2.93(dd,J=14.2,3.7 Hz,1H,H-18),2.23(t,J=13.5 Hz,1H,H-19 a ),2.13(s,3H),2.07(s,3H),2.03(s,3H),2.00(s,3H),1.98(s,3H),1.97-1.93(m,1H,H-2 a ),1.86-1.76(m,6H,H-11 ab ,H-22 a ,H-21 a ,H-22 b ),1.76-1.71(m,1H,H-15 a ),1.67-1.65(m,1H,H-2 b ),1.54(s,3H,イソプロピリデン C H 3 ),1.53-1.42(m,4H,H-9,H-1 a ,H-6 a ,H-7 a ),1.34-1.33(m,6H,H-27,イソプロピリデン C H3 ),1.31-1.24(m,9H,H-6′,H-6′′,H-6 b ,H-7 b ,H-15 b ), 1.11-1.10(m,1H,H-21 b ), 1.07-1.02(m,2H,H-19 b ),1.01-0.91(m,42H,H-23,H-30,TES C H 3 ),0.89(s,3H,H-25),0.88(s,5H,H-29),0.79(s,3H,H-24),0.73(s,3H,H-26),0.70-0.55(m,25H,H-5,TES C H 2 )ppm; HRMS(ESI-TOF)C 97 H 160 O 27 S 4 Na [M+Na] + The theoretical value was 1893.0144 and the measured value was 1893.0150. [ka] 3-O-(benzyl 2,3,4-tri-O-triethylsilyl-β-D-glucopyranosyluronate)-28-O-(2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl-(1→3)-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4))-2-O-acetyl-α-L-ramopyranosyl-(1→2)-3,4-O-isopropylidene-β-D-fucopyranosyl)-16-O-triethylsilyl chiral acid ester (44β)

[0138] 22 (22 mg, 0.019 mmol), 39 (23 mg, 0.019 mmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in (1 mL) 3 ·OEt 2 (approximately 48%, 2.0 μL, 0.0076 mmol) at -75 °C with N 2 The reaction was completed after 0.5 h when saturated NaHCO3 The reaction was quenched by the addition of and allowed to warm to room temperature. The resulting mixture was diluted with CH 2 Cl 2 Dilute with saturated NaHCO 3 , washed with brine, and MgSO 4 The residue was purified by column chromatography (silica gel; EtOAc / CH 2 Cl 2 / hexane = 4 / 1 / 7) to give 103 (44β: 19 mg, 46%; 44α: 2 mg, 5%) as a colorless syrup. f 0.60 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl 3 )δ 9.36(s,1H,H-23),7.38-7.30(m,5H),5.39(d,J=7.3 Hz,1H,H-1'),5.32(t,J=3.6 Hz,1H,H-12),5.17(dd,J=3.5,1.8 Hz,1H,H-2′′),5.16(d,J=1.8 Hz,2H,Bn C H 2 ),5.15-5.09(m,3H,H-3′′′,H-4′′′,H-3′′′′),5.02(m,J=7.8 Hz,1H,H-2′′′),4.95(d,J=1.8 Hz,1H,H-1′′),4.91(td,J=8.0,4.9 Hz,1H,H-4′′′′),4.85-4.81(m,2H,H-2′′′′,H-1′′′′),4.68(d,J=7.8 Hz,1H,H-1′′′),4.58(d,J=4.4 Hz,1H,H-1′′′′′′),4.47(s,1H,H-16),4.28-4.25(m,2H,H-4′′′′′′,H-6 a ′′′),4.21(d,J=6.3 Hz,1H,H-5′′′′′′),4.16(t,J=6.0 Hz,1H,H-3′),4.14-4.09(m,2H,H-6 b ",H-5 a′′′′),4.03(dd,J=6.0,1.9 Hz,1H,H-4′),3.99(dd,J=9.4,3.5 Hz,1H,H-3′′),3.86(qd,J=6.5,1.9 Hz,1H,H-5′),3.81(dd,J=11.8,4.6 Hz,1H,H-3),3.76(t,J=9.4 Hz,1H,H-4′′),3.71-3.62(m,3H,H-5′′,H-2′,H-5′′′),3.61(dd,J=5.4,1.2 Hz,1H,H-3′′′′′),3.57(dd,J=4.4,1.2 Hz,1H,H-2′′′′′),3.40(dd,J=11.9,8.1Hz,1H,H-5 b ′′′′),2.91(dd,J=14.4,4.2 Hz,1H,H-18),2.22(t,J=13.5 Hz,1H,H-19 a ),2.18(s,3H),2.11(s,3H),2.09(s,6H),2.04(s,3H),2.03(s,3H),2.01(s,3H),2.00(s,3H),1.99-1.97(m,1H,H-2 a ),1.87-1.84(m,2H,H-11 ab ),1.80(s,3H,H-21 a ,H-22 ab ),1.70-1.68(m,2H,H-2 b, H-15 a ),1.59-1.56(m,2H,H-9,H-1 a) ,1.54(s,3H,イソプロピリデン C H 3 ),1.45-1.41(m,2H,H-6 b ,H-7 a ),1.34(s,6H,H-27,イソプロピリデン C H 3 ),1.28(d,J=6.5 Hz,3H,H-6′),1.26-1.24(m,1H,H-15 b ),1.22(d,J=6.1Hz,3H,H-6′′),1.18-1.16(m,2H,H-5,H-6 b ),1.13-1.11(m,1H,H-21 b ),1.08(s,3H,H-24),1.06-1.02(m,1H,H-19 b), 1.00-0.92(m,44H,H-1 b ,H-7 b ,H-25,H-30,TES C H 3 ),0.88(s,3H,H-29),0.74(s,3H,H-26),0.68-0.55(m,24H,TES C H 2 )ppm; HRMS(ESI-TOF)C 109 H 174 O 36 S 4 Na [M+Na] + The theoretical value was 2195.0788 and the measured value was 2195.0785.

[0139] General procedure for amide bond formation: [ka]

[0140] A suspension of the benzyl ester starting material (1 equiv.) and 10% Pd / C (10% w / v) in THF (50 mM) was heated at room temperature for 2 h. 2 The reaction mixture was stirred under atmospheric pressure (balloon). The reaction mixture was stirred for 12-24 hours. The resulting mixture was filtered through Celite, concentrated, and dried under reduced pressure to give the crude acid intermediate. The crude acid intermediate, HBTU (3 equiv.) and DIPEA (3 equiv.) were dissolved in anhydrous CH 2 Cl 2 To a stirred solution of alkylamine (3 equiv.) in N 2 The mixture was stirred at room temperature under atmospheric pressure for 4 hours. The resulting mixture was concentrated and purified by column chromatography (silica gel, EtOAc / CH 2 Cl 2 The amide products were obtained in two-step yields of 61-95%.

[0141] In some embodiments, the silyl groups are randomly deprotected under hydrogenolysis conditions, resulting in an inseparable mixture in the subsequent amide coupling step. Therefore, a short column (silica, EtOAc / CH 2 Cl2 After filtration with hexane (hexane=1 / 1 / 2), the mixture is carried forward directly to the global deprotection step without structural characterization.

[0142] General procedure for overall deprotection: [ka]

[0143] Starting material CH 2 Cl 2 (10 mM) in pre-chilled TFA / H 2 O=4 / 1 solution (CH 2 Cl 2 (50% v / v relative to 100 mM NaOH) was added at 0° C. and stirred for 30 min. The solvent was evaporated under reduced pressure (<1 torr) at 0° C. and dried under high vacuum at room temperature for 1 h. A solution of the residue in MeOH (10 mM) was added with K 2 CO 3 (20 equiv.) was added and stirred at room temperature for 12 h. The suspension was centrifuged and the liquid was purified by HPLC to give the product in two-step yields of 30-75%. (HPLC column: SUPELCO Ascentis C18 25 cm × 10 mm, 5 μm; mobile phase: 20% ACN / H 2 O gradient ~ 90%ACN / H 2 O for 20 min, then 90% ACN / H 2 O isocratic for 15 min; flow rate: 5 mL / min). [ka] 3-O-(N-((dodecyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (46).

[0144] Following the general procedure for overall deprotection, 46 was obtained as a white solid in 68% yield: [α] D 20 -68.0(c 0.15,MeOH); 11H NMR (600 MHz, MeOD) δ 9.43 (s, 1H, H-23), 8.55 (s, 1H, amide N H ), 5.41 (d, J = 1.7 Hz, 1H, H-1′′), 5.31 (t, J = 3.5 Hz, 1H, H-12), 5.28 (d, J = 8.2 Hz, 1H, H-1′), 4.49 - 4.48 (m, 2H, H-1′′′, H-16), 4.26 (d, J = 7.8 Hz, 1H, H-1′′′′), 3.92 - 3.79 (m, 6H, H-2′′, H-3, H-5′′′, H-3′′, H-2′, H-5′′), 3.69 - 3.64 (m, 3H, H-5′, H-3′, H-5′′′′), 3.57 - 3.53 (m, 2H, H-4′, H-4′′), 3.49 - 3.42 (m, 2H, H-4′′′, H-4′′′′), 3.33 - 3 - 30 (m, 2H, H-3′′′, H-3′′′′), 3.28 - 3.26 (m, 1H, -NHC H a - ), 3.24 - 3.17 (m, 3H, -NHC H b - , H-2′′′, H-5′′′), 3.14 (dd, J = 9.2, 7.8 Hz, 1H, H-2′′′′), 2.94 (dd, J = 14.1, 4.3 Hz, 1H, H-18), 2.30 (t, J = 13.6 Hz, 1H, H-19a), 1.97 - 1.89 (m, 5H, H-2 a , H-11 ab , H-21 a , H-22 a ), 1.84 - 1.64 (m, 5H, H-2 b , H-22 b , H-9, H-1 a , H-15 a ), 1.58 - 1.48 (m, 4H, H-6 a , H-7 a , carbon chain C H 2 ), 1.45 (dd, J = 14.8, 2.7 Hz, 1H, H-15 b ), 1.40 (s, 3H, H-27), 1.37 - 1.27 (m, 23H, H-5, H-6 b , H-6′′, carbon chain C H 2 ×9), 1.22 (d, J = 6.4 Hz, 3H, H-6′), 1.19 - 1.16 (m, 1H, H-21 b ), 1.14 (s, 3H, H-24), 1.12 - 1.09 (m, 1H, H-1 b ), 1.07 - 1.04 (m, 1H, H-19 b ), 1.02 (s, 3H, H-25), 0.98 - 0.94 (m, 4H, H-7b, H-30), 0.91 (t, J = 7.0 Hz, 3H, carbon chain C H 3 ), 0.88 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13 C NMR (151 MHz, MeOD) δ 209.2 (C-23), 177.2 (C-28), 171.5, 144.9 (C-13), 123.1 (C-12), 107.0 (C-1′′′), 105.1 (C-1′′′′), 101.1 (C-1′′), 95.2 (C-1′), 84.0 (C-4′′), 83.5 (C-3), 78.2 (C-3′′′), 77.6 (C-3′′′′), 76.7 (C-3′), 76.6 (C-5′′′′), 76.1 (C-2′′′), 74.7 (C-2′′′′), 74.6 (C-16), 74.0 (C-2′), 73.6 (C-4′), 73.2 (C-4′′′′), 72.7 (C-5′′), 72.2 (C-3′′), 71.9 (C-2′′), 71.1 (C-4′′′), 68.8 (C-5′), 67.3 (C-5′′′), 56.1 (C-4), 50.0 (C-17), 49.6 (C-5), 48.1 (C-9), 48.0 (C-19), 42.8 (C-14), 42.4 (C-18), 41.1 (C-8), 40.1 (carbon chain C H 2 ), 39.5 (C-1), 37.1 (C-10), 36.5 (C-21), 36.5 (C-15), 33.6 (C-6), 33.4 (C-29), 33.1 (carbon chain C H 2 ), 32.0 (C-22), 31.3 (C-20), 30.9 (carbon chain C H 2 ), 30.9 (carbon chain C H 2 ), 30.8 (carbon chain C H 2 ), 30.8 (carbon chainC H 2 ), 30.6 (carbon chain C H 2 ), 30.5 (carbon chain C H 2 ), 30.3 (carbon chain C H 2 ), 28.0 (carbon chain C H 2 ),27.2(C-27),26.0(C-2),24.8(C-30),24.5(C-11),23.8(Carbon chain C H 2 ),21.6(C-7),18.3(C-6′′),17.7(C-26),16.5(C-6′),16.4(C-25),14.5(Carbon chain C H 3 ),10.6(C-24)ppm; HRMS(ESI-TOF)C 65 H 107 NO 22 Na [M+Na] + The theoretical value was 1276.7177 and the measured value was 1276.7209. [ka] 3-O-(N-(methyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (47).

[0145] Following the general procedure for overall deprotection, 47 was obtained as a white solid in 36% yield: [α] D 20 -26.8(c 0.22,MeOH); 1 H NMR (600 MHz, MeOD) δ 9.42 (s, 1H, H-23), 8.52 (s, 1H, amide N H),5.40(d,J=1.7 Hz,1H,H-1′′),5.31(t,1H,J=3.5 Hz,H-12),5.28(d,J=8.2 Hz,1H,H-1′),4.49-4.47(m,2H,H-1′′′,H-16),4.24(d,J=7.9 Hz,1H,H-1′′′′),3.90(dd,J=3.3,1.7 Hz,1H,H-2′′),3.89-3.79(m,5H,H-3,H-5 a ′′′,H-3′′,H-2′,H-5′′),3.69-3.64(m,3H,H-5′,H-3′,H-5′′′′),3.56-3.53(m,2H,H-4′,H-4′′),3.46(ddd,J=10.4,8.8,5.4 Hz,1H,H-4′′′),3.41(t,J=9.1Hz. 1H,H-4′′′′),3.33-3.30(m,2H,H-3′′′,H-3′′′′)3.23-3.17(m,2H,H-2′′′,H-5 b ′′′),3.13(dd,J=9.2,7.9 Hz,1H,H-2′′′′),2.94(dd,J=14.6,4.1Hz,1H,H-18),2.80(s,3H,-NHC H 3 ),2.30(t,J=13.7 Hz,1H,H-19 a ),1.94-1.91(m,5H,H-2 a ,H-11 ab ,H-21 a ,H-22 a ),1.81-1.66(m,5H,H-2 b ,H-22 b ,H-9,H-1 a ,H-15 a ),1.55-1.43(m,3H,H-6 a ,H-7 a ,H-15 b ),1.40(s,3H,H-27),1.35-1.31(m,5H,H-5,H-6 b ,H-6′′),1.22(d,J=6.4 Hz,3H,H-6′),1.18(d,J=10.3,2.5 Hz,1H,H-21 b ),1.13(s,3H,H-24),1.10(dd,J=13.2,4.02 Hz,1H,H-1 b), 1.05 (dd, J = 13.4, 3.5 Hz, 1H, H-19 b ), 1.02 (s, 3H, H-25), 0.97 - 0.95 (m, 4H, H-7 b , H-30), 0.88 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13 C NMR (151 MHz, MeOD) δ 209.3 (C-23), 177.2 (C-28), 172.3, 144.8 (C-13), 123.2 (C-12), 107.0 (C-1′′′), 104.8 (C-1′′′′), 101.1 (C-1′′), 95.2 (C-1′), 84.1 (C-4′′), 83.3 (C-3), 78.2 (C-3′′′), 77.5 (C-3′′′′), 76.7 (C-3′), 76.5 (C-5′′′′), 76.1 (C-2′′′), 74.7 (C-2′′′′), 74.6 (C-16), 74.0 (C-2′), 73.6 (C-4′), 73.4 (C-4′′′′), 72.7 (C-5′), 72.2 (C-3′′), 71.9 (C-2′′), 71.1 (C-4′′′), 68.8 (C-5′′), 67.3 (C-5′′′), 56.1 (C-4), 50.0 (C-17), 49.6 (C-5), 48.0 (C-9), 48.0 (C-19), 42.8 (C-14), 42.4 (C-18), 41.1 (C-8), 39.3 (C-1), 37.1 (C-10), 36.5 (C-21), 36.5 (C-15), 33.6 (C-6), 33.4 (C-29), 32.0 (C-22), 31.3 (C-20), 27.2 (C-27), 26.2 (-NH C H 3 ), 25.8 (C-2), 24.8 (C-30), 24.5 (C-11), 21.5 (C-7), 18.3 (C-6′′), 17.7 (C-26), 16.5 (C-6′), 16.3 (C-25), 10.5 (C-24) ppm; HRMS (ESI-TOF) C 54 H 86 NO 22 [M + H] + The theoretical value at [M + H] was 1100.5636, and the measured value was 1100.5667. [Chemical formula] 3-O-(N-(hexyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (48).

[0146] Following the general procedure for overall deprotection, 48 was obtained as a white solid in 45% yield: [α] D 20 -17.35(c 0.34,MeOH); 1 H NMR (600 MHz, MeOD) δ 9.42 (s, 1H, H-23), 8.55 (s, 1H, amide N H ),5.40(d,J=1.6 Hz,1H,H-1′′),5.31(t,J=3.4 Hz,1H,H-12),5.28(d,J=8.2 Hz,1H,H-1′),4.49-4.48(m,2H,H-16,H-1′′′),4.26(d,J=7.9 Hz,1H,H-1′′′′),3.92-3.77(m,6H,H-2′′,H-3,H-5 a ′′′,H-3′′,H-2′,H-5′′),3.70-3.62(m,3H,H-5′,H-3′,H-5′′′′),3.56-3.53(m,2H,H-4′,H-4′′) ,3.49-3.41(m,2H,H-4′′′,H-4′′′′),3.33-3.30(m,2H,H-3′′′,H-3′′′′),3.29-3.17(m,4H,-NHC H 2 - ,H-2′′′,H-5 b ′′′),3.14(dd,J=9.2,7.9 Hz,1H,H-2′′′′),2.94(dd,J=14.1,4.0 Hz,1H,H-18),2.30(t,J=13.8 Hz,1H,H-19 a ), 1.96-1.90(m,5H,H-2 a ,H-11 ab ,H-21 a ,H-22 a ), 1.84-1.64(m,5H,H-2 b ,H-22 b ,H-9,H-1 a ,H-15 a), 1.55 - 1.49 (m, 4H, H-6 a , H-7 a , carbon chain C H 2 ), 1.45 (dd, J = 14.8, 2.7 Hz, 1H, H-15 b ), 1.40 (s, 3H, H-27), 1.36 - 1.29 (m, 13H, H-5, H-6 b , H-6′′, carbon chain C H 2 ×4), 1.22 (d, J = 6.4 Hz, 3H, H-6′), 1.18 - 1.17 (m, 1H, H-21 b ), 1.13 (s, 3H, H-24), 1.12 - 1.09 (m, 1H, H-1 b ), 1.05 (dd, J = 12.6, 3.0 Hz, 1H, H-19 b ), 1.01 (s, 3H, H-25), 0.95 (s, 4H, H-7 b , H-30), 0.92 (t, J = 7.0 Hz, 3H, carbon chain C H 3 ), 0.88 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13 C NMR (151 MHz, MeOD) δ 209.3 (C-23), 177.2 (C-28), 171.5, 144.8 (C-13), 123.2 (C-12), 106.9 (C-1′′′), 104.9 (C-1′′′′), 101.1 (C-1′′), 95.2 (C-1′), 84.0 (C-4′′), 83.3 (C-3), 78.2 (C-3′′′), 77.6 (C-3′′′′), 76.7 (C-3′), 76.6 (C-5′′′′), 76.1 (C-2′′′), 74.7 (C-2′′′′), 74.6 (C-16), 74.1 (C-2′), 73.6 (C-4′), 73.2 (C-4′′′′), 72.7 (C-5′), 72.2 (C-3′′), 71.9 (C-2′′), 71.1 (C-4′′′), 68.8 (C-5′′), 67.3 (C-5′′′), 56.1 (C-4), 50.0 (C-17), 49.6 (C-5), 48.0 (C-9, C-19), 42.8 (C-14), 42.4 (C-18), 41.1 (C-8), 40.1 (carbon chain C H 2),39.4(C-1),37.1(C-10),36.5(C-21),36.5(C-15),33.6(C-6),33.4(C-29),32.8(Carbon chain C H 2 ),32.0(C-22),31.3(C-20),30.3(carbon chain C H 2 ), 27.6 (carbon chain C H 2 ),27.2(C-27),25.9(C-2),24.8(C-30),24.5(C-11),23.8(Carbon chain C H 2 ),21.5(C-7),18.3(C-6′′),17.7(C-26),16.5(C-6′),16.3(C-25),14.5(Carbon chain C H 3 ),10.6(C-24)ppm; HRMS(ESI-TOF)C 59 H 96 NO 22 [M+H] + The theoretical value was 1170.6418 and the measured value was 1170.6448. [ka] 3-O-(N-(octadecyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (49).

[0147] Following the general procedure for global deprotection (HPLC column: Alltima C8 150 mm × 4.6 mm, 5 μm, flow rate: 1 mL / min), a white solid 49 was obtained in 30% yield: [α] D 20 -27.4 (c 0.27, MeOH); 1 H NMR (600 MHz, MeOD) δ 9.43 (s, 1H, H-23), 8.55 (s, 1H, amide N H), 5.41 (d, J = 1.6 Hz, 1H, H-1′′), 5.31 (t, J = 3.4 Hz, 1H, H-12), 5.28 (d, J = 8.2 Hz, 1H, H-1′), 4.49 (d, J = 7.6 Hz, 2H, H-16, H-1′′′), 4.26 (d, J = 7.9 Hz, 1H, H-1′′′′), 3.92 - 3.77 (m, 6H, H-2′′, H-3, H-5 a ′′′, H-3′′, H-2′, H-5′′), 3.69 - 3.63 (m, 3H, H-5′, H-3′, H-5′′′′), 3.57 - 3.53 (m, 2H, H-4′, H-4′′), 3.49 - 3.41 (m, 2H, H-4′′′, H-4′′′′), 3.33 - 3.27 (m, 3H, H-3′′′, H-3′′′′, -NHC H a - ), 3.24 - 3.17 (m, 3H, H-2′′′, H-5 b ′′′, -NHC H b - ), 3.14 (dd, J = 9.2, 7.9 Hz, 1H, H-2′′′′), 2.94 (dd, J = 14.3, 4.1 Hz, 1H, H-18), 2.30 (t, J = 13.6 Hz, 1H, H-19 a ), 1.98 - 1.89 (m, 5H, H-2 a , H-11 ab , H21 a , H-22 a) , 1.84 - 1.64 (m, 5H, H-2 b , H-9, H-22 b , H-1 a , H-15 a ), 1.55 - 1.49 (m, 4H, H-6 a , H-7 a , carbon chain C H 2 ), 1.45 (dd, J = 14.8, 2.5 Hz, 1H, H-15 b ), 1.40 (s, 3H, H-27), 1.37 - 1.25 (m, 35H, H-5, H-6 b , H-6′′, carbon chain C H 2 ×15), 1.22 (d, J = 6.4 Hz, 3H, H-6′), 1.20 - 1.15 (m, 1H, H-21b ), 1.14 (s, 3H, H-24), 1.13 - 1.08 (m, 1H, H-1 b ), 1.06 (dd, J = 12.6, 3.0 Hz, 1H, H-19 b ), 1.02 (s, 3H, H-25), 0.97 - 0.95 (m, 4H, H-7 b , H-30), 0.90 (t, J = 7.0 Hz, 3H, carbon chain C H 3 ), 0.88 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13 C NMR (151 MHz, MeOD) δ 209.2 (C-23), 177.2 (C-28), 171.5, 144.9 (C-13), 123.1 (C-12), 107.0 (C-1′′′), 105.1 (C-1′′′′), 101.1 (C-1′′), 95.7 (C-1′), 84.0 (C-4′′), 83.5 (C-3), 78.2 (C-3′′′), 77.6 (C-3′′′′), 76.7 (C-3′), 76.6 (C-5′′′′), 76.1 (C-2′′′), 74.7 (C-2′′′′), 74.6 (C-16), 74.0 (C-2′), 73.6 (C-4′), 73.2 (C-4′′′′), 72.7 (C-5′), 72.2 (C-3′′), 71.9 (C-2′′), 71.1 (C-4′′′), 68.7 (C-5′′), 67.3 (C-5′′′), 56.1 (C-4), 50.0 (C-17), 49.6 (C-5), 48.1 (C-9), 48.0 (C-19), 42.8 (C-14), 42.3 (C-18), 41.1 (C-8), 40.0 (carbon chain C H 2 ), 39.5 (C-1), 37.1 (C-10), 36.5 (C-21), 36.5 (C-15), 33.6 (C-6), 33.4 (C-29), 33.1 (carbon chain C H 2 ), 32.0 (C-22), 31.4 (C-20), 30.9 (carbon chain C H 2 ), 30.9 (carbon chain C H 2 ), 30.8 (carbon chain C H 2 ), 30.8 (carbon chain C H 2),30.6(carbon chain C H 2 ),30.5(carbon chain C H 2 ),30.3(carbon chain C H 2 ),28.0(carbon chain C H 2 ),27.2(C-27),26.0(C-2),24.9(C-30),24.5(C-11),23.8(Carbon chain C H 2 ),21.6(C-7),18.3(C-6′′),17.7(C-26),16.5(C-6′),16.4(C-25),14.5(Carbon chain C H 3 ),10.6(C-24)ppm; HRMS(ESI-TOF)C 71 H 120 NO 22 [M+H] + The theoretical value was 1338.8297 and the measured value was 1338.3327. [ka] 3-O-(N-(8-(4-Methoxyphenyl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (53).

[0148] Following the general procedure for overall deprotection, 53 was obtained as a white solid in 54% yield: [α] D 20 -123.3(c 0.06,MeOH); 1 H NMR (600 MHz, MeOD) δ 9.42 (s, 1H, H-23), 8.55 (s, 1H, amide N H), 7.07 (d, J = 8.7 Hz, 2H), 6.82 (d, J = 8.7 Hz, 2H), 5.41 (d, J = 1.8 Hz, 1H, H-1′′), 5.30 - 5.27 (m, 2H, H-12, H-1′), 4.49 - 4.48 (m, 2H, H-16, H-1′′′), 4.26 (d, J = 7.8 Hz, 1H, H-1′′′′), 3.91 (dd, J = 3.3, 1.8 Hz, 1H, H-2′′), 3.90 - 3.79 (m, 5H, H-3, H-5 a ′′′, H-3′′, H-2′, H-5′′), 3.76 (s, 3H, OC H 3 ), 3.69 - 3.66 (m, 2H, H-5′, H-3′), 3.64 (d, J = 9.7 Hz, 1H, H-5′′′′), 3.58 - 3.53 (m, 2H, H-4′, H-4′′), 3.49 - 3.42 (m, 2H, H-4′′′, H-4′′′′), 3.33 - 3 - 30 (m, 2H, H-3′′′, H-3′′′′), 3.28 - 3.25 (m, 1H, -NHC H a - ), 3.24 - 3.17 (m, 3H, H-2′′′, H-5 b ′′′, -NHC H b - ), 3.14 (dd, J = 9.2, 7.8 Hz, 1H, H-2′′′′), 2.94 (dd, J = 14.3, 4.1 Hz, 1H, H-18), 2.54 (t, J = 7.4 Hz 2H, carbon chain C H 2 Ph), 2.30 (t, J = 13.6 Hz, 1H, H-19 a ), 1.97 - 1.88 (m, 5H, H-2 a , H-11 ab , H-21 a , H-22 a ), 1.84 - 1.72 (m, 3H, H-2 b , H-9, H-22 b ), 1.71 - 1.66 m, 2H, H-1 a , H-15 a ), 1.62 - 1.47 (m, 6H, carbon chain C H 2 ×2, H-6 a , H-7 a), 1.47 - 1.43 (m, 1H, H-15 b ), 1.40 (s, 3H, H-27), 1.37 - 1.30 (m, 13H, H-5, H-6 b , H-6′′, carbon chain C H 2 ×4), 1.22 (d, J = 6.4 Hz, 3H, H-6′), 1.18 - 1.16 (m, 1H, H-21 b ), 1.13 (s, 3H, H-24), 1.10 - 1.08 (m, 1H, H-1 b ), 1.05 (dd, J = 12.1, 3.6 Hz, 1H, H-19 b ), 1.00 (s, 3H, H-25), 0.97 - 0.94 (m, 4H, H-7 b , H-30), 0.87 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13 C NMR (151 MHz, MeOD) δ 209.2 (C-23), 177.2 (C-28), 171.5, 159.2, 144.8 (C-13), 135.9, 130.3, 123.1 (C-12), 114.8, 107.0 (C-1′′′), 105.0 (C-1′′′′), 101.1 (C-1′′), 95.2 (C-1′), 84.0 (C-4′′), 83.5 (C-3), 78.2 (C-3′′′), 77.6 (C-3′′′′), 76.7 (C-3′), 76.6 (C-5′′′′), 76.1 (C-2′′′), 74.7 (C-2′′′′), 74.6 (C-16), 74.0 (C-2′), 73.6 (C-4′), 73.2 (C-4′′′′), 72.7 (C-5′), 72.2 (C-3′′), 71.9 (C-2′′), 71.1 (C-4′′′), 68.8 (C-5′′), 67.3 (C-5′′′), 56.1 (C-4), 55.7 (O C H 3 ), 50.0 (C-17), 49.6 (C-5), 48.1 (C-19), 48.0 (C-9), 42.8 (C-14), 42.3 (C-18), 41.1 (C-8), 40.0 (carbon chain C H 2 ), 39.4 (C-1), 37.1 (C-10), 36.5 (C-21), 36.5 (C-15), 36.7 (carbon chain C H 2),33.6(C-6),33.4(C-29),33.0(carbon chain C H 2 ),32.0(C-22),31.3(C-20),30.7(carbon chain C H 2 ), 30.5 (carbon chain C H 2 ), 30.3 (carbon chain C H 2 ), 30.2 (carbon chain C H 2 ), 27.9 (carbon chain C H 2 ),27.2(C-27),26.0(C-2),24.8(C-30),24.5(C-11),21.6(C-7),18.3(C-6′′),17.7(C-26),16.5(C-6′),16.4(C-25),10.6(C-24)ppm; HRMS(ESI-TOF)C 68 H 106 NO 23 [M+H] + The theoretical value was 1326.6970 and the measured value was 1326.6978. [ka] 3-O-(N-(8-(4-fluorophenyl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (54).

[0149] Following the general procedure for overall deprotection, 54 was obtained as a white solid in 56% yield: [α] D 20 -28.3(c 0.18,MeOH); 1 H NMR (600 MHz, MeOD) δ 9.42 (s, 1H, H-23), 8.55 (s, 1H, amide N H), 7.17 (dd, J = 8.8, 5.4 Hz, 2H), 6.98 (t, J = 8.8 Hz, 2H), 5.41 (d, J = 1.7 Hz, 1H, H-1′′), 5.29 - 5.27 (m, 2H, H-12, H-1′), 4.49 - 4.48 (m, 2H, H-16, H-1′′′), 4.26 (d, J = 7.9 Hz, 1H, H-1′′′′), 3.92 - 3.78 (m, 6H, H-2′′, H-3, H-5 a ′′′, H-3′′, H-2′, H-5′′), 3.70 - 3.63 (m, 3H, H-5′, H-3′, H-5′′′′), 3.57 - 3.52 (m, 2H, H-4′, H-4′′), 3.48 - 3.41 (m, 2H, H-4′′′, H-4′′′′), 3.33 - 3.30 (m, 2H, H-3′′′, H-3′′′′), 3.28 - 3.25 (m, 1H, -NHC H a - ), 3.24 - 3.16 (m, 3H, H-2′′′, H-5 b ′′′, -NHC H b - ), 3.13 (dd, J = 9.2, 7.9 Hz, 1H, H-2′′′′), 2.94 (dd, J = 14.3, 4.5 Hz, 1H, H-18), 2.59 (t, J = 7.6 Hz, 2H, carbon chain C H 2 Ph), 2.29 (t, J = 13.6 Hz, 1H, H-19 a ), 1.96 - 1.89 (m, 5H, H-2 a , H-11 ab , H-21 a , H-22 a ), 1.83 - 1.72 (m, 3H, H-2 b , H-9, H-22 b ), 1.72 - 1.64 (m, 2H, H-1 a , H-15 a ), 1.63 - 1.58 (m, 2H, carbon chain C H 2 ), 1.56 - 1.48 (m, 4H, H-6 a , H-7 a ), 1.45 (dd, J = 14.9, 2.6 Hz, 1H, H-15 b), 1.40 (s, 3H, H-27), 1.36 - 1.31 (m, 13H, H-5, H-6 b , H-6′′, carbon chain C H 2 × 4), 1.22 (d, J = 6.4 Hz, 3H, H-6′), 1.18 - 1.16 (m, 1H, H-21 b ), 1.13 (s, 3H, H-24), 1.11 - 1.08 (m, 1H, H-1 b ), 1.05 - 1.03 (m, 1H, H-19 b ), 1.00 (s, 3H, H-25), 0.96 - 0.94 (m, 4H, H-7 b , H-30), 0.86 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13 C NMR (151 MHz, MeOD) δ 209.2 (C-23), 177.2 (C-28), 171.5, 162.6 (J = 241Hz), 144.9 (C-13), 139.8, 130.9 (J = 8 Hz), 123.1 (C-12), 115.8 (J = 21Hz), 107.0 (C-1′′′), 105.0 (C-1′′′′), 101.1 (C-1′′), 95.2 (C-1′), 84.0 (C-4′′), 83.4 (C-3), 78.2 (C-3′′′), 77.6 (C-3′′′′), 76.7 (C-3′), 76.6 (C-5′′′′), 76.1 (C-2′′′), 74.7 (C-2′′′′), 74.6 (C-16), 74.0 (C-2′), 73.6 (C-4′), 73.2 (C-4′′′′), 72.7 (C-5′), 72.2 (C-3′′), 71.9 (C-2′′), 71.1 (C-4′′′), 68.8 (C-5′′), 67.3 (C-5′′′), 56.1 (C-4), 50.0 (C-17), 49.6 (C-5), 48.1 (C-9), 48.0 (C-19), 42.8 (C-14), 42.4 (C-18), 41.1 (C-8), 40.0 (carbon chain C H 2 ), 39.4 (C-1), 37.1 (C-10), 36.5 (C-21), 36.5 (C-15), 36.1 (carbon chain C H 2),33.6(C-6),33.4(C-29),32.9(C-),32.0(C-22),31.3(C-20),30.7(Carbon chain C H 2 ), 30.5 (carbon chain C H 2 ), 30.3 (carbon chain C H 2 ), 30.3 (carbon chain C H 2 ), 30.2 (carbon chain C H 2 ), 27.9 (carbon chain C H 2 ),27.2(C-27),26.0(C-2),24.8(C-30),24.5(C-11),21.5(C-7),18.3(C-6′′),17.7(C-26),16.5(C-)6′,16.4(C-25),10.6(C-24)ppm; HRMS(ESI-TOF)C 67 H 102 FNO 22 Na [M+Na] + The theoretical value was 1314.6770 and the measured value was 1314.6794. [ka] 3-O-(N-(8-(4-fluorophenyl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (55).

[0150] Following the general procedure for global deprotection, 55 was obtained as a white solid in 75% yield: [α] D 20 -32.5(c 0.24,MeOH); 1 H NMR (600 MHz, MeOD) δ 9.42 (s, 1H, H-23), 8.55 (s, 1H, amide N H), 7.29 (d, J = 8.3 Hz, 2H), 7.08 (d, J = 8.3 Hz, 2H), 5.41 (d, J = 1.7 Hz, 1H, H-1′′), 5.31 (t, J = 3.6 Hz, 1H, H-12), 5.29 (d, J = 8.2 Hz, 1H, H-1′), 4.49 - 4.48 (m, 2H, H-1′′′, H-16), 4.26 (d, J = 7.8 Hz, 1H, H-1′′′′), 3.91 - 3.78 (m, 6H, H-2′′, H-3, H-5 a ′′′, H-3′′, H-2′, H-5′′), 3.69 - 3.63 (m, 3H, H-5′, H-3′, H-5′′′′), 3.57 - 3.53 (m, 2H, H-4′, H-4′′), 3.48 - 3.42 (m, 2H, H-4′′′, H-4′′′′), 3.33 - 3.30 (m, 2H, H-3′′′, H-3′′′′), 3.29 - 3.25 (m, 1H, -NHC H a - ), 3.23 - 3.17 (m, 3H, H-2′′′, H-5 b ′′′, -NHC H a - ), 3.14 (dd, J = 9.2, 7.8 Hz, 1H, H-2′′′′), 2.94 (dd, J = 14.5, 4.3 Hz, 1H, H-18), 2.57 (m, J = 7.6 Hz, 2H, carbon chain C H 2 Ph), 2.30 (t, J = 13.6 Hz, 1H, H-19 a ), 1.97 - 1.89 (m, 5H, H-2 a , H-11 ab , H-21 a , H-22 a ), 1.83 - 1.72 (m, 3H, H-2 b , H-9, H-22 b ), 1.72 - 1.64 (m, 2H, H-1 a , H-15 a ), 1.63 - 1.57 (m, 2H, carbon chain C H 2 ), 1.56 - 1.47 (m, 4H, H-6 a , H-7 a , carbon chain C H 2), 1.47 - 1.43 (m, 1H, H-15 b ), 1.40 (s, 3H, H-27), 1.36 - 1.28 (m, 22H, H-5, H-6 b , H-6′′, carbon chain C H 2 ×4, tBu C H 3 ×3), 1.22 (d, J = 6.4 Hz, 3H, H-6′), 1.19 - 1.15 (m, 1H, H-21 b ), 1.13 (s, 3H, H-24), 1.11 - 1.08 (m, 1H, H-1 b ), 1.08 - 1.03 (m, 1H, H-19 b ), 1.00 (s, 3H, H-25), 0.96 - 0.94 (m, 4H, H-7 b , H-30), 0.86 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13 C NMR (151 MHz, MeOD) δ 209.2 (C-23), 177.2 (C-28), 171.5, 149.5, 144.8 (C-13), 140.8, 129.1, 126.1, 123.1 (C-12), 107.0 (C-1′′′), 105.0 (C-1′′′′), 101.1 (C-1′′), 95.2 (C-1′), 84.1 (C-4′′), 83.5 (C-3), 78.2 (C-3′′′), 77.6 (C-3′′′′), 76.7 (C-3′), 76.6 (C-5′′′′), 76.1 (C-2′′′), 74.7 (C-2′′′′), 74.6 (C-16), 74.0 (C-2′), 73.6 (C-4′), 73.2 (C-4′′′′), 72.7 (C-5′), 72.2 (C-3′′), 71.9 (C-2′′), 71.1 (C-4′′′), 68.8 (C-5′′), 67.1 (C-5′′′), 56.1 (C-4), 50.0 (C-17), 49.6 (C-5), 48.1 (C-9), 48.0 (C-19), 42.8 (C-14), 42.4 (C-18), 41.1 (C-8), 40.0 (carbon chain C H 2 ), 39.4 (C-1), 37.1 (C-10), 36.4 (C-15, C-21, carbon chain C H 2),35.2(tBu 4° C),33.6(C-6),33.4(C-29),32.8(Carbon chain C H 2 ), 31.9(C-22,tBu C H 3 ×3),31.3(C-20),30.7(carbon chain C H 2 ), 30.5 (carbon chain C H 2 ), 30.4 (carbon chain C H 2 ), 30.2 (carbon chain C H 2 ), 27.9 (carbon chain C H 2 ),27.2(C-27),26.0(C-2),24.8(C-30),24.5(C-11),21.6(C-7),18.3(C-6′′),17.7(C-26),16.5(C-6′),16.4(C-25),10.6(C-24)ppm; HRMS(ESI-TOF)C 71 H 111 NO 22 Na [M+Na] + The theoretical value was 1352.7490 and the measured value was 1352.7515. [ka] 3-O-(N-(8-(4-phenoxyphenyl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (56).

[0151] Following the general procedure for overall deprotection, 56 was obtained as a white solid in 46% yield: [α] D 20 -71.4(c 0.07,MeOH); 1 H NMR (600 MHz, MeOD) δ 9.42 (s, 1H, H-23), 8.55 (s, 1H, amide N H), 7.32 (dd, J = 8.6, 7.5 Hz, 2H), 7.17 (dd, J = 8.6, 1.0 Hz, 2H), 7.07 (tt, J = 7.5, 1.0 Hz, 1H), 6.95 (d, J = 8.6 Hz, 2H), 6.91 (d, J = 8.6 Hz, 2H), 5.41 (d, J = 1.7 Hz, 1H, H-1′′), 5.30 (s, 1H, H-12), 5.27 (d, J = 8.2 Hz, 1H, H-1′), 4.49 - 4.47 (m, 2H, H-16, H-1′′′), 4.26 (d, J = 7.8 Hz, 1H, H-1′′′′), 3.91 - 3.79 (m, 6H, H-2′′, H-3, H-5 a ′′′, H-3′′, H-2′, H-5′′), 3.67 - 3.63 (m, 3H, H-5′, H-3′, H-5′′′′), 3.56 - 3.53 (m, 2H, H-4′, H-4′′), 3.49 - 3.42 (m, 2H, H-4′′′, H-4′′′′), 3.33 - 3.25 (m, 3H, H-3′′′, H-3′′′′, -NHC H a - ), 3.23 - 3.17 (m, 3H, H-2′′′, H-5 b ′′′, -NHC H b - ), 3.14 (dd, J = 9.2, 7.8 Hz, 1H, H-2′′′′), 2.94 (dd, J = 14.2, 4.4 Hz, 1H, H-18), 2.60 (t, J = 7.5 Hz, 2H, carbon chain C H 2 Ph), 2.29 (t, J = 13.1Hz, 1H, H-19 a ), 1.98 - 1.87 (m, 5H, H-2 a , H-11 ab , H-21 a , H-22 a ), 1.82 - 1.76 (m, 3H, H-2 b , H-9, H-22 b ), 1.71 - 1.65 (m, 2H, H-1 a , H-15 a ), 1.64 - 1.60 (m, 2H, carbon chain C H 2 ), 1.56 - 1.48 (m, 4H, H-6 a , H-7 a), 1.45 (dd, J = 14.9, 2.6 Hz, 1H, H-15 b ), 1.40 (s, 3H, H-27), 1.37 - 1.31 (m, 13H, H-5, H-6 b , H-6′′, carbon chain C H 2 ), 1.21 (d, J = 6.4 Hz, 3H, H-6′), 1.18 - 1.15 (m, 1H, H-21 b ), 1.13 (s, 3H, H-24), 1.11 - 1.08 (m, 1H, H-1 b ), 1.06 - 1.03 (m, 1H, H-19 b ), 1.00 (s, 3H, H-25), 0.97 - 0.93 (m, 4H, H-7 b , H-30), 0.85 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13 C NMR (151 MHz, MeOD) δ 209.2 (C-23), 177.1 (C-28), 171.5, 159.2, 156.5, 144.9 (C-13), 139.2, 130.8, 130.7, 124.0, 123.1 (C-12), 120.0, 119.4, 107.0 (C-1′′′), 105.0 (C-1′′′′), 101.1 (C-1′′), 95.2 (C-1′), 84.1 (C-4′′), 83.5 (C-3), 78.2 (C-3′′′), 77.6 (C-3′′′′), 76.7 (C-3′), 76.6 (C-5′′′′), 76.1 (C-2′′′), 74.7 (C-2′′′′), 74.6 (C-16), 74.0 (C-2′), 73.6 (C-4′), 73.2 (C-4′′′′), 72.7 (C-5′), 72.2 (C-3′′), 71.9 (C-2′′), 71.1 (C-4′′′), 68.7 (C-5′′), 67.3 (C-5′′′), 56.1 (C-4), 50.0 (C-17), 49.6 (C-5), 48.1 (C-9), 48.0 (C-19), 42.8 (C-14), 42.4 (C-18), 41.1 (C-8), 40.0 (carbon chain C H 2 ), 39.4 (C-1), 37.1 (C-10), 36.5 (C-21), 36.5 (C-15), 36.2 (carbon chain C H 2),33.6(C-6),33.4(C-29),32.9(carbon chain C H 2 ),32.0(C-22),31.3(C-20),30.7(carbon chain C H 2 ), 30.5 (carbon chain C H 2 ), 30.4 (carbon chain C H 2 ), 30.3 (carbon chain C H 2 ), 27.9 (carbon chain C H 2 ),27.2(C-27),26.0(C-2),24.8(C-30),24.5(C-11),21.6(C-7),18.3(C-6′′),17.7(C-26),16.5(C-6′),16.4(C-25),10.6(C-24)ppm; HRMS(ESI-TOF)C 73 H 107 NO 23 Na [M+Na] + The theoretical value was 1388.7126 and the measured value was 1388.7172. [ka] 3-O-(N-(4-phenoxyphen-1-yl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (57). Following the general procedure for overall deprotection, 57 was obtained as a white solid in 23% yield: 1 H NMR (600 MHz, CD 3OD)δ 9.45(s,1H),8.55(s,1H),7.61(d,J=8.9 Hz,2H),7.34(t,J=7.7 Hz,2H),7.09(t,J=7.1Hz,1H),7.00-6.95(m,4H),5.41(d,J=1.6 Hz,1H),5.30(t,J=3.6 Hz,1H),5.28(d,J=8.2 Hz,1H),4.50-4.47(m,2H),4.32(d,J=7.8 Hz,1H),3.93(dd,J=11.8,4.6 Hz,1H),3.90(dd,J=3.2,1.8 Hz,1H),3.87 - 3.77(m,5H),3.67-3.65(m,2H),3.60-3.53(m,3H),3.46(ddd,J=10.4,9.0,5.4 Hz,1H),3.36(t,J=9.0 Hz,1H),3.33-3.30(m,1H)3.24-3.17(m,3H),2.93(dd,J=14.4,4.3 Hz,1H),2.29(t,J=13.6 Hz,1H),2.00-1.90(m,5H),1.86-1.64(m,5H),1.61-1.48(m,2H),1.45(dd,J=14.8,2.6 Hz,1H),1.39(s,3H),1.37-1.33(m,2H),1.31(d,J=6.2 Hz,3H),1.21(d,J=6.4 Hz,3H),1.20-1.16(m,1H),1.15(s,3H),1.13-1.08(m,1H),1.06-1.02(m,1H),1.01(s,3H),0.97-0.94(m,4H),0.87(s,3H),0.77(s,3H); 13 C NMR(151 MHz,CD 3OD)δ 209.4,177.2,170.3,169.4,159.0,155.2,144.7,134.8,130.9,124.3,123.2,120.3,119.5, 106.9,105.2,101.1,95.2,84.0,83.4,78.2,77.6,77.6,77.6,76.7,76.1,74.7,74.6,74.0,7 3.6,73.0,72.7,72.2,71.9,71.1,68.7,67.3,56.2,50.0,49.6,48.0,48.0,42.8,42.3,41.1, 39.3,37.1,36.5,36.5,36.5,33.4,32.0,31.3,27.2,24.8,21.6,18.3,17.7,16.5,16.3,10.6 ppm; HRMS(ESI-TOF)C 65 H 92 NO 23 [M+H] + The theoretical value was 1254.6055 and the measured value was 1254.6060. [ka] 3-O-(N-(2-(4-phenoxyphen-1-yl)ethyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (58). Following the general procedure for overall deprotection, 58 was obtained as a white solid in 58% yield: 1 H NMR (600 MHz, CD 3OD)δ 9.40(s,1H),8.55(s,1H),7.33(dd,J=8.5,7.5 Hz,2H),7.23(d,J=8.5 Hz,2H),7.09(t,J=7.5 Hz,1H),6.95(d,J=8.5 Hz,2H),6.93(d,J=8.6 Hz,2H),5.41(d,J=1.7 Hz,1H),5.30-5.27(m,2H),4.50-4.47(m,2H),4.23(d,J=7.9 Hz,1H),3.91(dd,J=3.2,1.7 Hz,1H),3.87-3.77(m,5H),3.70-3.65(m,2H),3.63(d,J=9.7 Hz,1H),3.57-3.50(m,3H),3.50-3.43(m,2H),3.40(t,J=9.3 Hz,1H),3.33-3.29(m,2H),3.23-3.17(m,2H),3.11(dd,J=9.2,7.9 Hz,1H),2.94(dd,J=14.3,4.1Hz,1H),2.83(t,J=6.8 Hz,2H),2.30(t,J=13.6 Hz,1H),1.98 - p1.79(m,8H),1.77(dd,J=13.8,4.3 Hz,1H),1.74-1.64(m,4H),1.58-1.41(m,3H),1.39(s,3H),1.35(d,J=11.7 Hz,1H),1.34-1.28(m,5H),1.22(d,J=6.4 Hz,3H),1.18(d,J=12.8 Hz,1H),1.11(s,3H),1.08-1.02(m,2H),0.97(s,3H),0.96-0.92(m,4H),0.89(s,3H),0.76(s,3H); BBD 13C NMR(151 MHz,MeOD)δ 209.3,177.2,171.7,159.0,157.0,144.7,135.6,131.3,130.9,124.2,123.2,120.2,119.5,106. 9,104.9,101.1,95.2,84.0,83.5,78.2,77.5,76.7,76.4,76.1,74.7,74.6,74.0,73.6,73.4,72.7 ,72.2,71.9,71.1,68.7,67.3,56.1,50.0,49.6,48.0,48.0,42.8,42.3,41.5,41.1,39.4,37.0,3 6.5,36.5,35.4,33.6,33.4,32.0,31.3,27.2,25.9,24.8,24.6,21.5,18.3,17.7,16.5,16.3,10.6 ppm; HRMS(ESI-TOF)C 67 H 96 NO 23 [M+H] + The theoretical value was 1282.6368 and the measured value was 1282.6370. [ka] 3-O-(N-(16-(4-phenoxyphen-1-yl)hexadecyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester. (59) Following the general procedure for global deprotection (HPLC column: Alltima C8 150 mm×4.6 mm, 5 μm, flow rate: 1 mL / min), a white solid 59 was obtained in 34% yield: 1 H NMR (600 MHz, CD 3OD)δ 9.42(s,1H),8.55(s,1H),7.32(dd,J=8.5,7.5 Hz,2H),7.16(d,J=8.5 Hz,2H),7.07(t,J=7.5 Hz,1H),6.94(d,J=8.5 Hz,2H),6.89(d,J=8.5 Hz,2H),5.41(d,J=1.6 Hz,1H),5.31(brs,1H),5.28(d,J=8.2 Hz,1H),4.50 - 4.47(m,2H),4.26(d,J=7.8 Hz,1H),3.92-3.78(m,6H),3.68-3.63(m,3H),3.57-3.53(m,2H),3.48-3.42(m,2H),3.35-3.27(m,3H),3.24-3.16(m,3H),3.14(dd,J=9.2,7.8 Hz,1H),2.97-2.92(m,1H),2.60(t,J=7.7 Hz,2H),2.30(t,J=13.6 Hz,1H),1.97-1.90(m,5H),1.83-1.65(m,5H),1.63-1.59(m,2H),1.56-1.43(m,5H),1.40(s,3H),1.38-1.27(m,29H),1.21(d,J=6.4 Hz,3H),1.17(d,J=12.6 Hz,1H),1.13(s,3H),1.09(d,J=13.4 Hz,1H),1.07-1.03(m,1H),1.01(s,3H),0.97-0.92(m,4H),0.87(s,3H),0.77(s,3H); BBD 13 C NMR(151 MHz,CD 3OD)δ 209.2,177.1,171.5,159.2,156.4,144.9,139.3,130.8,130.7,124.0,123.1,112.0,119.4,106.9,105.1,101.1,9 5.2,84.0,83.5,78.2,77.8,76.7,76.7,76.6,76.1,74.7,74.6,73.9,73.6,73.2,72.7,72.2,71.9,71.2,68.7,67.3 ,56.1,50.0,49.6,48.1,48.0,42.8,42.3,41.1,40.0,37.1,36.5,36.5,36.2,33.6,33.4,32.8,31.4,30.9,30.9,3 0.9,30.8,30.8,30.7,30.7,30.6,30.6,30.5,30.3,28.0,27.2,26.0,24.9,24.5,21.6,18.3,17.7,16.5,16.4,10.6 ppm; HRMS(ESI-TOF)C 81 H 124 NO 23 [M+H] + The theoretical value was 1478.8559 and the measured value was 1478.8560. [ka] 3-O-(N-(8-(4-(4-fluorophenoxy)phen-1-yl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (60). Following the general procedure for amide bond formation and global deprotection, 60 was obtained as a white solid in 40% yield: 1 H NMR (600 MHz, CD 3OD)δ 9.42(s,1H),7.16(d,J=8.5 Hz,2H),7.09-7.04(m,2H),6.99-6.95(m,2H),6.88(d,J=8.5 Hz,2H),5.41(d,J=1.6 Hz,1H),5.29-5.27(m,2H),4.50-4.48(m,2H),4.26(d,J=7.8 Hz,1H),3.92-3.77(m,6H),3.67-3.63(m,3H),3.57-3.52(m,2H),3.48-3.43(m,2H),3.33-3.30(m,2H),3.29-3.26(m,1H),3.23-3.17(m,3H),3.14(dd,J=9.2,7.8 Hz,1H),2.93(dd,J=14.3,4.1Hz,1H),2.60(t,J=7.6 Hz,2H),2.29(t,J=13.6 Hz,1H),1.97-1.88(m,5H),1.83-1.64(m,5H),1.63-1.59(m,2H),1.57-1.48(m,4H),1.45(dd,J=14.7,2.4 Hz,1H),1.39(s,3H),1.38-1.30(m,13H),1.21(d,J=6.4 Hz,3H),1.19-1.15(m,1H),1.13(s,3H),1.11-1.07(m,1H),1.04(dd,J=13.6,3.7 Hz,1H),0.99(s,3H),0.96(d,J=11.6 Hz,1H),0.92(s,3H),0.85(s,3H),0.76(s,3H); 13 C NMR(151 MHz,CD 3OD)δ 209.2,177.1,171.5,160.8,159.2,156.9,155.1,144.9,139.1,130.8,123.1,121.2,121.2,119.5,117.3,117. 1,106.9,105.1,101.0,95.2,84.0,83.5,78.2,77.6,76.7,76.6,76.1,74.7,74.6,73.9,73.6,73.2,72.7,72.2, 71.9,71.1,68.7,67.3,56.1,50.0,49.6,48.1,48.0,42.8,42.3,41.1,40.0,39.4,37.1,36.5,36.5,36.2,33.6 ,33.4,32.9,32.0,31.3,30.7,30.5,30.4,30.2,27.9,27.2,26.0,24.8,24.5,21.5,18.3,17.7,16.5,16.4,10.6 ppm; HRMS(ESI-TOF)C 73 H 107 FNO 23 [M+H] + The theoretical value was 1384.7212 and the measured value was 1384.7224. [ka] 3-O-(N-(8-(4-hydroxyphen-1-yl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (61). Following the general procedure for overall deprotection, 61 was obtained as a white solid in 23% yield: 1 H NMR (600 MHz, CD 3OD)δ 9.42(s,1H),8.55(s,1H),6.97(d,J=8.5 Hz,2H),6.69(d,J=8.5 Hz,2H),5.41(d,J=1.7 Hz,1H),5.30(t,J=3.1Hz,1H),5.29(d,J=8.2 Hz,1H),4.49(d,J=7.7 Hz,2H),4.26(d,J=7.8 Hz,1H),3.93-3.77(m,6H),3.69-3.63(m,3H),3.58-3.52(m,2H),3.49-3.41(m,2H),3.33-3.25(m,3H),3.24-3.16(m,3H),3.14(dd,J=9.2,7.8 Hz,1H),2.94(dd,J=14.2,4.2 Hz,1H),2.50(t,J=7.5 Hz,2H),2.30(t,J=13.6 Hz,1H),1.98-1.88(m,5H),1.83-1.64(m,5H),1.60-1.48(m,6H),1.47-1.43(m,1H),1.40(s,3H),1.37-1.27(m,13H),1.22(d,J=6.4 Hz,3H),1.18-1.16(m,1H),1.13(s,3H),1.11-1.03(m,2H),1.00(s,3H),0.96-0.95(m,4H),0.87(s,3H),0.77(s,3H); BBD 13 C NMR(151 MHz,CD 3 OD)δ 209.2,177.2,171.5,156.3,144.8,134.8,130.3,123.2,116.1,106.9,105.0,101.1,95.2,84.0,83.4,78.2,77.6,76.7,76.6,76.1,74.7,74.6,74.0,73.6,73.2,72.7,72.2,71.9,71.1,68.7,67.3,56.1,50.0,49.6,48.0,42.8,42.3,41.1,40.0,39.4,37.1,36.5,36.5,36.1,33.6,33.4,33.1,32.0,31.3,30.7,30.5,30.3,30.3,27.9,27.2,26.0,24.8,24.5,21.5,18.3,17.7,16.5,16.4,10.6 ppm; HRMS(ESI-TOF)C 67 H 104 NO 23[M+H] + The theoretical value was 1290.6994 and the measured value was 19690.7008. [ka] 3-O-(N-(8-(4-morpholinophen-1-yl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (62). Following the general procedure for overall deprotection, 62 was obtained as a white solid in 43% yield: 1 H NMR(600 MHz,MeOD)δ 9.42(s,1H),8.55(s,1H),7.07(d,J=8.5 Hz,2H),6.90(d,J=8.5 Hz,2H),5.41(d,J=1.1Hz,1H),5.31-5.27(m,2H),4.49(d,J=7.6 Hz,1H),4.48(brs,1H)4.26(d,J=7.8 Hz,1H),3.92-3.90(m,1H),3.89 - 3.77(m,9H),3.69-3.63(m,3H),3.57-3.52(m,2H),3.49-3.42(m,2H),3.33-3.26(m,3H),3.24-3.16(m,3H),3.13(dd,J=9.2,7.8 Hz,1H),3.11-3.08(m,4H),2.94(dd,J=14.3,4.0 Hz,1H),2.53(t,J=7.6 Hz,2H),2.29(t,J=13.6 Hz,1H),1.98-1.86(m,5H),1.82-1.63(m,5H),1.62-61.47(m,6H),1.47-1.43(m,1H),1.40(s,3H),1.37-1.29(m,13H),1.22(d,J=6.4 BBD 13 C NMR (151 MHz, CD 3OD)δ 209.2,177.1,171.5,150.9,144.8,135.9,130.0,123.1,117.4,106.9,105.1,101.1,95.1,84.0,83.6 ,78.2,77.6,76.7,76.6,76.1,74.7,74.6,74.0,73.6,73.2,72.7,72.2,71.9,71.1,68.7,68.0,67.3,5 6.1,51.4,50.0,49.6,48.1,48.0,42.8,42.3,41.1,40.0,39.4,37.1,36.5,36.5,36.1,33.6,33.4,33 .0,32.0,31.3,30.8,30.5,30.3,30.2,27.9,27.2,26.0,24.9,24.1,21.5,18.3,17.8,16.5,16.4,10.6 ppm; HRMS(ESI-TOF)C 71 H 111 N 2 O 23 [M+H] + The theoretical value was 1359.7572 and the measured value was 1359.7580. [ka] 3-O-(N-(8-(furan-2-yl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (66). Following the general procedure for overall deprotection, 66 was obtained as a white solid in 23% yield: 1 H NMR (600 MHz, CD 3OD)δ 9.42(s,1H),8.55(s,1H),7.33(d,J=1.1Hz,1H),6.30-6.26(m,1H),5.99(d,J=3.1Hz,1H),5.41(d,J=1.4 Hz,1H),5.30-5.27(m,2H),4.50-4.47(m,2H),4.26(d,J=7.8 Hz,1H),3.93-3.76(m,6H),3.71-3.62(m,3H),3.58-3.50(m,2H),3.48-3.42(m,2H),3.33-3.25(m,3H),3.23-3.16(m,3H),3.13(dd,J=9.0,7.8 Hz,1H),2.94(dd,J=14.3,4.2 Hz,1H),2.62(d,J=7.5 Hz,2H),2.31(d,J=13.4 Hz,1H),1.99-1.87(m,5H),1.84-1.59(m,7H),1.59-1.48(m,4H),1.45(dd,J=14.8,2.5 Hz,1H),1.40(s,3H),1.37-1.30(m,13H),1.22(d,J=6.4 Hz,3H),1.18(d,J=11.0 Hz,1H),1.13(s,3H),1.12-1.08(m,1H),1.07-1.03(m,1H),1.01(s,3H),0.97-1.94(m,4H),0.87(s,3H),0.77(s,3H); BBD 13 C NMR(151 MHz,CD 3 OD)δ 209.2,177.2,171.5,157.5,144.8,141.9,123.2,111.0,106.9,105.7,105.0,101.1,95.2,84.0,83.4,78.2,77.6,76.7,76.6,76.1,74.7,74.6,74.0,73.6,73.2,72.7,72.2,71.9,71.1,68.7,67.3,56.1,50.0,49.8,48.0,42.8,42.3,41.1,40.0,39.4,37.1,36.5,36.5,33.6,33.4,32.0,31.3,30.6,30.5,30.2,29.3,28.9,27.9,27.2,26.0,24.8,24.5,21.5,18.3,17.7,16.5,16.3,10.6 ppm; HRMS(ESI-TOF)C 65 H 102NO 23 [M+H] + The theoretical value was 1264.6837 and the measured value was 1264.6846. [ka] 3-O-(N-(8-(pyridin-3-yl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (68).

[0152] Following the general procedure for amide bond formation and global deprotection, 68 was obtained as a white solid in 42% yield: 1 H NMR (600 MHz, CD 3 OD)δ 9.42(s,1H),8.55(s,1H),8.38(d,J=1.8 Hz,1H),8.36(dd,J=4.8,1.8 Hz,1H),7.70(dt,J=7.8,1.8 Hz,1H),7.37(dd,J=7.7,4.8 Hz,1H),5.41(d,J=1.6 Hz,1H),5.29-5.27(m,2H),4.50-4.47(m,2H),4.26(d,J=7.8 Hz,1H),3.92-3.77(m,6H),3.69-3.63(m,3H),3.56-3.53(m,2H),3.49- 3.42(m,2H),3.33-3.25(m,3H),3.23-3.17(m,3H),3.14(dd,J=9.2,7.8 Hz,1H),2.93(dd,J=14.3,4.2 Hz,1H),2.67(t,J=7.6 Hz,2H),2.29(t,J=13.6 Hz,1H),1.98-1.87(m,5H),1.83-1.62(m,7H),1.56-1.48(m,4H),1.45(dd,J=14.7,2.4 Hz,1H),1.39(s,3H),1.38-1.28(m,13H),1.22(d,J=6.4 Hz,3H),1.19-1.15(m,1H),1.13(s,3H),1.10(d,J=13.5 Hz,1H),1.06-1.01(m,1H),1.00(s,3H),0.96-0.94(m,4H),0.86(s,3H),0.76(s,3H);13 C NMR (151 MHz, CD 3 OD)δ 209.2,177.2,171.5,170.3,150.0,147.5,144.8,140.3,138.3,130.9,125.2,123.1,106.9,105.0,101.1,9 5.2,84.08,83.4,78.2,77.6,76.7,76.6,76.1,74.7,74.6,74.0,73.6,73.2,72.7,72.2,71.9,71.2,68.7,6 7.3,56.1,50.0,49.6,48.1,48.0,42.8,42.3,41.1,40.0,39.4,37.1,36.5,36.4,33.8,33.6,33.4,32.4,32 .0,31.3,30.8,30.8,30.6,30.4,30.3,30.2,27.9,27.2,26.0,24.8,24.5,21.5,18.3,17.7,16.5,16.4,10.6 ppm; HRMS(ESI-TOF)C 66 H 103 N 2 O 22 [M+H] + The theoretical value was 1275.6997 and the measured value was 1275.7031. [ka] 3-O-(N-(8-(4-phenoxyphenyl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-α-D-fucopyranosyl) chiral acid ester (56α).

[0153] Following the general procedure for overall deprotection, 56α was obtained as a white solid in 47% yield: 1 H NMR (600 MHz, MeOD) δ 9.41 (s, 1H, H-23), 8.55 (s, 1H, amide N H), 7.32 (dd, J = 8.6, 7.5 Hz, 2H), 7.16 (d, J = 8.6 Hz, 2H), 7.07 (t, J = 7.4 Hz, 1H), 6.95 (d, J = 8.5 Hz, 2H), 6.90 (d, J = 8.5 Hz, 2H), 6.06 (d, J = 3.7 Hz, 1H, H-1′), 5.33 (t, J = 3.4 Hz, 1H, H-12), 4.87 (d, J = 1.7 Hz, 1H, H-1′′) 4.49 (s, 1H, H-16), 4.42 (d, J = 7.7 Hz, 1H, H-1′′′), 4.26 (d, J = 7.8 Hz, 1H, H-1′′′′), 3.97 (dd, J = 3.2, 1.7 Hz, 1H, H-2′′), 3.92 - 3.86 (m, 3H, H-3, H-2′, H-5′), 3.85 - 3.80 (m, 2H, H-3′, H-5 a ′′′), 3.77 (dd, J = 9.4, 3.2 Hz, 1H, H-3′′), 3.72 (d, J = 3.0 Hz, 1H, H-4′), 3.65 (d, J = 9.7 Hz, 1H, H-5′′′′), 3.53 (dq, J = 9.4, 6.0 Hz, 1H, H-5′′), 3.49 (t, J = 9.4 Hz, 1H, H-4′′), 3.45 - 3.41 (m, 2H, H-4′′′, H-4′′′′), 3.33 - 3.25 (m, 3H, H-3′′′, H-3′′′′, -NHC H a - ), 3.23 - 3.16 (m, 3H, H-2′′′, H-5 b ′′′, -NHC H b - ), 3.14 (dd, J = 9.2, 7.8 Hz, 1H, H-2′′′′), 2.98 (dd, J = 14.3, 4.1Hz, 1H, H-18), 2.60 (t, J = 7.7 Hz, 2H, carbon chain C H 2 Ph), 2.26 (t, J = 13.7 Hz, 1H), 2.01 - 1.88 (m, 5H, H-2 a , H-11 ab , H-21 a , H-22 a ), 1.83 - 1.74 (m, 3H, H-2 b , H-22 b , H-9), 1.70 (d, J = 13.4 Hz, 1H, H-1a ), 1.65 - 1.59 (m, 3H, H-15 a , carbon chain C H 2 ), 1.58 - 1.47 (m, 4H, H-6 a , H-7 a , carbon chain C H 2 ), 1.41 - 1.38 (m, 4H, H-15 b , H-27), 1.37 - 1.31 (m, 9H, H-5, carbon chain C H 2 ×4), 1.24 (d, J = 6.0 Hz, 3H, H-6′′), 1.23 - 1.19 (m, 2H, H-6 b , H-21 b ), 1.17 (d, J = 6.5 Hz, 3H, H-6′), 1.12 (s, 3H, H-24), 1.11 - 1.08 (m, H-1 b ), 1.05 (dd, J = 11.7, 4.3 Hz, 1H, H-19 b ), 1.01 (s, 3H, H-25), 0.95 - 0.91 (m, 4H, H-7 b , H-30), 0.87 (s, 3H, H-29), 0.79 (s, 3H, H-26); 1313C NMR (151 MHz, MeOD) δ 209.1 (C-23), 176.8 (C-28), 171.5, 159.2, 156.5, 144.7 (C-13), 139.1, 130.8, 130.7, 124.0, 123.4 (C-12), 120.0, 119.4, 107.1 (C-1‴), 105.0 (C-1⁗), 104.5 (C-1″), 93.0 (C-1′), 84.5 (C-4″), 83.3 (C-3), 78.4 (C-3‴), 77.6 (C-3⁗), 76.9 (C-2′), 76.6 (C-5⁗), 76.1 (C-2‴), 75.0 (C-16), 74.7 (C-2⁗), 73.5 (C-4′), 73.2 (C-4⁗), 72.5 (C-3″), 71.6 (C-2″), 71.0 (C-4‴), 71.0 (C-3′), 70.5 (C-5′), 68.8 (C-5″), 67.2 (C-5‴), 56.1 (C-4), 50.6 (C-17), 49.6 (C-5), 48.0 (C-9), 48.0 (C-19), 42.8 (C-14), 42.1 (C-18), 41.0 (C-8), 40.0, 39.4 (C-1), 37.1 (C-10), 36.3 (C-21, C-15), 36.2, 33.6 (C-6), 33.4 (C-29), 32.9, 32.4 (C-22), 31.4 (C-20), 30.7, 30.4, 30.3, 30.2, 27.9, 27.2 (C-27), 26.0 (C-2), 25.3 (C-30), 24.5 (C-11), 21.4 (C-7), 18.0 (C-26), 17.9 (C-6″), 16.8 (C-6′), 16.3 (C-25), 10.5 (C-29) ppm; HRMS (ESI-TOF) C 73 H 108 NO 23 [M + H] + The theoretical value in was 1366.7307 and the measured value was 1366.7318.

Chemical Structure

[0154] Following the general procedure for overall deprotection, 77β was obtained as a white solid in 67% yield: 1 H NMR (600 MHz, MeOD) δ 9.42 (s, 1H, H-23), 8.55 (s, 1H, amide N H ),7.32(dd,J=8.5,7.4 Hz,2H),7.16(dd,J=8.5,1.0 Hz,2H),7.07(tt,J=7.6,1.0 Hz,1H),6.95(d,J=8.5 Hz,2H),6.90(d,J=8.5 Hz,2H),5.61(d,J=3.7 Hz,1H,H-1′),5.35(t,J=3.5 Hz,1H,H-12),5.02(s,1H,H-1′′),4.51(d,J=7.7 Hz,1H,H-1′′′),4.49(s,1H,H-16),4.26(d,J=7.8 Hz,1H,H-1′′′′),3.92-3.79(m,7H,H-5 a ′,H-3,H-3′,H-2′′,H-3′′,H-5 a ′′′,H-4′),3.77(dd,J=5.2,3.7 Hz,1H,H-2′),3.73-3.67(m,1H,H-5′′),3.65(d,J=9.7 Hz,1H,H-5′′′′),3.57(t,J=9.1Hz,1H,H-4′′),3.50-3.41(m,3H,H-5 b ′,H-4′′′,H-4′′′′),3.33-3.26(m,3H,H-3′′′,H-3′′′′,-NHC H a - ),3.23-3.16(m,3H,H-2′′′,H-5 b ''',-NHC H b -), 3.14 (dd, J = 9.2, 7.8 Hz, 1H, H-2′′′′), 3.05 (dd, J = 14.3, 4.2 Hz, 1H, H-18), 2.60 (t, J = 7.6 Hz, 2H, carbon chain C H 2 Ph), 2.28 (t, J = 13.6 Hz, 1H, H-19 a ), 1.97 - 1.87 (m, 5H, H-2 a , H-11 ab , H-21 a , H-22 a ), 1.84 - 1.66 (m, 5H, H-2 a , H-22 b , H-9, H-15 a , H-1 a ), 1.66 - 1.59 (m, 2H, carbon chain C H 2 ), 1.59 - 1.50 (m, 3H, H-6a, carbon chain C H 2 ), 1.42 - 1.37 (m, 4H, H-15 b , H-27), 1.36 - 1.32 (m, 9H, H-5, carbon chain C H 2 ×4), 1.31 - 1.27 (m, 5H, H-6 b , H-7 a , H-6′′), 1.17 - 1.12 (m, 4H, H-21 b , H-24), 1.12 - 1.07 (m, 1H, H-1 b ), 1.04 (dd, J = 12.7, 3.1 Hz, 1H, H-19 b ), 1.00 (s, 3H, H-25), 0.97 - 0.92 (m, 4H, H-30, H-7 b ), 0.86 (s, 3H, H-29), 0.77 (s, 3H, H-26); 13C NMR(151 MHz,MeOD)δ 209.1(C-23),176.9(C-28),171.5,159.2,156.5,144.9(C-13),139.1,130.8,130.7,124.0,123.4(C-12),120.0,119.4,106.6(C-1′′′) ,105.0(C-1′′′′),101.3(C-1′′),94.0(C-1′),83.4(C-3),83.3(C-4 ′′),78.1(C-3′′′),77.6(C-3′′′′),76.6(C-5′′′′),76.0(C-2′′′), 75.5(C-2′),74.7(C-2′′′′),74.6(C-16),73.2(C-4′′′′),72.3(C-3′′),72.1(C-2′′),71.1(C-3′,C-4′′′),69.0(C-5′′),67.2(C-5′′′) ),67.0(C-4′),63.7(C-5′),56.1(C-4),50.2(C-17),49.6(C-5),48.1(C-9),47.7(C-19),42.8(C-14),42.1(C-18),41.1(C-8),40.0(carbon lock C H 2 ),39.4(C-1),37.1(C-10),36.4(C-21),36.3(C-15),36.2(Carbon lock C H 2 ),33.5(C-6),33.4(C-29),32.9(Carbon lock C H 2 ),32.0(C-22),31.4(C-20),30.7(Carbon lock C H 2 ),30.5(carbon lock C H 2 ),30.3(carbon lock C H 2 ),30.3(carbon lock C H 2 ),27.9(carbon lock C H 2 ),27.3(C-27),26.0(C-2),25.1(C-30),24.5(C-11),21.5(C-7),18.1(C-6′′),17.9(C-26),16.4(C-25),10.6(C-24)ppm; HRMS(ESI-TOF)C 72 H 105NO 23 [M+H] + The theoretical value was 1352.7150 and the measured value was 1352.7167. [ka] 3-O-(N-(8-(4-phenoxyphenyl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl) chiral acid ester (77α).

[0155] Following the general procedure for overall deprotection, 77α was obtained as a white solid in 35% yield: 1 H NMR (600 MHz, MeOD) δ 9.41 (s, 1H, H-23), 8.55 (s, 1H, amide N H ),7.32(dd,J=8.6,7.4 Hz,2H),7.16(d,J=8.6 Hz,2H),7.07(t,J=7.4 Hz,1H),6.95(d,J=8.6 Hz,2H),6.90(d,J=8.6 Hz,2H),6.09(d,J=3.6 Hz,1H,H-1′),5.34(t,J=3.5 Hz,1H,H-12),4.89(d,J=1.7 Hz,H-1′′)4.49(s,1H,H-16),4.43(d,J=7.7 Hz,1H,H-1′′′),4.26(d,J=7.8 Hz,1H,H-1′′′′),3.96(dd,J=3.1,1.7 Hz,1H,H-2′′),3.94(dd,J=10.1,3.6 Hz,1H,H-2′),3.92-3.90(m,H-4′),3.89(dd,J=11.8,4.7 Hz,1H,H-3),3.85(dd,J=10.1,3.3 Hz,1H,H-3′),3.83(dd,J=11.4,5.4 Hz,1H,H-5 a ′′′), 3.80-3.76(m,2H,H-5 a ′,H-3′′),3.67(dd,J=12.4,1.8 Hz,1H,H-5 b′), 3.65 (d, J = 9.7 Hz, 1H, H - 5′′′′), 3.58 - 3.53 (m, 1H, H - 5′′), 3.49 (t, J = 9.4 Hz, 1H, H - 4′′), 3.46 - 3.41 (m, 2H, H - 4′′′, H - 4′′′′), 3.33 - 3.26 (m, 3H, H - 3′′′, H - 3′′′′, -NHC H a - ), 3.23 - 3.12 (m, 4H, H - 2′′′, H - 5 b ′′′, H - 2′′′′, -NHC H b - ), 2.98 (dd, J = 14.4, 4.2 Hz, 1H, H - 18), 2.60 (t, J = 7.6 Hz, 2H, carbon chain C H 2 Ph), 2.26 (t, J = 13.6 Hz, 1H, H - 19 a ), 2.00 - 1.87 (m, 5H, H - 2 a , H - 11 ab , H - 21 a , H - 22 a ), 1.83 - 1.78 (m, 2H, H - 2 b , H - 22 b ), 1.75 (dd, J = 10.9, 6.7 Hz, 1H, H - 9), 1.73 - 1.65 (m, 2H, H - 1 a , H - 15 a ), 1.65 - 1.60 (m, 2H, carbon chain C H 2 ), 1.60 - 1.46 (m, 4H, H - 6 a , H - 7 a , carbon chain C H 2 ), 1.43 - 1.38 (m, 4H, H - 15 b , H - 27), 1.37 - 1.32 (m, 9H, H - 5, carbon chain C H 2 ×4), 1.26 - 1.24 (m, 4H, H - 6 b , H - 6′′), 1.21 (d, J = 12.5 Hz, 1H, H - 21 b ), 1.12 (s, 3H, H - 24), 1.10 (dd, J = 13.7, 3.6 Hz, 1H, H - 1 b),1.05(dd,J=12.4,3.7 Hz,1H,H-19 b ),1.01(s,3H,H-25),0.95-0.91(m,4H,H-7 b ,H-30),0.87(s,3H,H-29),0.77(s,3H,H-26); 13 C NMR(151 MHz,MeOD)δ 209.0(C-23),176.8(C-28),171.5,170.3,159.2,156.5,144.5(C-13) ,139.2,130.8,130.7,124.0,123.5(C-12),120.0,119.4,107.1(C-1′ ′′),105.0(C-1′′′′),104.3(C-1′′),93.5(C-1′),84.4(C-4′′),83.3 (C-3),78.4(C-3′′′),77.6(C-3′′′′),77.0(C-2′),76.6(C-5′′′′),76 .1(C-2′′′),74.9(C-16),74.7(C-2′′′′),73.2(C-4′′′′),72.5(C-3′ ′),71.6(C-2′′),71.0(C-4′′′),70.7(C-4′),70.1(C-3′),68.8(C-5′′ ),67.2(C-5′′′),66.5(C-5′),56.1(C-4),50.7(C-17),49.6(C-5),48 .0(C-9),47.9(C-19),42.8(C-14),42.2(C-18),41.1(C-8),40.0(Carbon Lock C H 2 ),39.4(C-1),37.1(C-10),36.4(C-21),36.3(C-15),36.2(Carbon lock C H 2 ),33.7(C-6),33.3(C-29),32.9(Carbon lock C H 2 ),32.3(C-22),31.4(C-20),30.7(Carbon lock C H 2 ),30.4(Carbon lock C H 2 ),30.3(Carbon lock C H 2 ),30.2(Carbon lock C H 2 ),27.9(Carbon lock CH 2 ),27.2(C-27),26.0(C-2),25.3(C-30),24.5(C-11),21.4(C-7),17.9(C-6′′),17.9(C-26),16.3(C-25),10.5(C-24)ppm; HRMS(ESI-TOF)C 72 H 106 NO 23 [M+H] + The theoretical value was 1352.7150 and the measured value was 1352.7159. [ka] 3-O-(N-(8-(4-phenoxyphenyl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) echinocyst ester (78).

[0156] Following the general procedure for overall deprotection, 78 was obtained as a white solid in 69% yield: 1 H NMR(600 MHz,MeOD) δ 8.55(s,1H,amide N H ),7.32(dd,J=8.6,7.4 Hz,2H),7.16(dd,J=8.6,1.0 Hz,2H),7.06(tt,J=7.4,1.0 Hz,1H),6.95(d,J=8.6 Hz,2H),6.90(d,J=8.6 Hz,2H),5.41(d,J=1.7 Hz,1H,H-1′′),5.30-5.27(m,2H,H-12,H-1′),4.51-4.48(m,2H,H-1′′′,H-16),4.40(d,J=7.8 Hz,1H,H-1′′′′),3.92(dd,J=3.3,1.7 Hz,1H,H-2′′),3.88-3.79(m,4H,H-5 a′′′, H-3′′, H-2′, H-5′′), 3.69 - 3.62 (m, 3H, H-5′′′′, H-3′, H-5′), 3.58 - 3.53 (m, 2H, H-4′′, H-4′), 3.50 - 3.45 (m, 2H, H-4′′′, H-4′′′′), 3.38 (t, J = 9.1 Hz, 1H, H-3′′′′), 3.35 - 3.28 (m, 2H, H-3′′′, -NHC H a - ), 3.27 - 3.24 (m, 2H, H-2′′′, H-2′′′′), 3.23 - 3.17 (m, 3H, H-3, H-5 b ′′′, -NHC H b - ), 2.93 (dd, J = 14.3, 4.2 Hz, 1H, H-18), 2.60 (m, J = 7.5 Hz, 2H, carbon chain C H 2 Ph), 2.29 (t, J = 13.6 Hz, 1H, H-19 a ), 1.96 - 1.82 (m, 5H, H-2 a , H-11 ab , H-21 a , H-22 a ), 1.80 - 1.66 (m, 3H, H-22 b , H-2 b , H-15 a ), 1.66 - 1.55 (m, 5H, H-1 a , H-7 a , H-9, carbon chain C H 2 ), 1.55 - 1.49 (m, 3H, H-6 a , carbon chain C H 2 ), 1.48 - 1.41 (m, 2H, H-15 b , H-6 b ), 1.40 - 1.36 (m, 4H, H-7b, H-27), 1.36 - 1.32 (m, 11H, H-6′′, carbon chain C H 2 ×4), 1.21 (d, J = 6.4 Hz, 3H, H-6′), 1.16 (dd, J = 10.8, 3.8 Hz, 1H, H-21 b),1.06(s,3H,H-23),1.03(dd,J=12.1,8.9 Hz,1H,H-19 b ),0.98(dd,J=13.4,3.5 Hz,1H,H-1 b ),0.95(s,3H,H-25),0.92(s,3H,H-30),0.86(s,3H,H-24),0.85(s,3H,H-29),0.78(d,J=12.0 Hz,1H,H-5),0.76(s,3H,H-26); 13 C NMR(151 MHz,MeOD)δ 177.1(C-28),171.7,159.2,156.5,144.8(C-13),139.1,130.8,130.7,124.0,123.4(C-12),120.1,119.4,107.0(C-1′′′),106.8(C-1′′′′),101.1(C-1′′),95.2(C-1′),91.0(C-3),84.1(C-4′′),78.1(C-3′′′),77.8(C-3′′′′),76.7(C-3′),76.5(C-5′′′′),76.1(C-2′′′),75.2(C-2′′′′),74.7(C-16),74.0(C-2′),73.6(C-4′),73.4(C-4′′′′),72.7(C-5′),72.2(C-3′′),71.9(C-2′′),71.1(C-4′′′),68.7(C-5′′),67.3(C-5′′′),57.2(C-5),50.0(C-17),48.1(C-9),48.1(C-19),42.7(C-14),42.3(C-18),40.8(C-8),40.2(C-4),40.0(C-1),39.9,37.9(C-10),36.5(C-21),36.5(C-15),36.4,34.3(C-6),33.4(C-29),32.9,32.0(C-22),31.3(C-20),30.7,30.6,30.4,30.3,28.5(C-23),27.9,27.3(C-2),27.2(C-27),24.8(C-30),24.6(C-11),19.4(C-7),18.3(C-6′′),17.8(C-26),17.0(C-24),16.5(C-6′),16.3(C-25)ppm; HRMS(ESI-TOF)C 73 H 110 NO 22[M+H] + The theoretical value was 1352.7514 and the measured value was 1352.7532. [ka] 3-O-(N-(8-(4-phenoxyphenyl)octyl)-β-D-glucopyranosyluronamide)-28-O-(β-D-glucopyranosyl-(1→3)-(β-D-xylopyranosyl-(1→4))-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (79).

[0157] Following the general procedure for global deprotection, 79 was obtained as a white solid in 33% yield: 1H NMR (600 MHz, MeOD) δ 9.42 (s, 1H, H-23), 8.55 (s, 1H, amide N H ),7.32(dd,J=8.5,7.5 Hz,2H),7.17(d,J=8.5 Hz,2H),7.07(t,J=7.5 Hz,1H),6.95(d,J=8.5 Hz,2H),6.90(d,J=8.5 Hz,2H),5.29(s,1H,H-12),5.26(d,J=8.1Hz,1H,H-1′),5.23(s,1H,H-1′′),4.70(d,J=7.9 Hz,1H,H-1′′′′),4.53(d,J=7.3 Hz,1H,H-1′′′),4.46(s,1H,H-16),4.27-4.25(m,2H,H-2′′,H-1′′′′′),3.95(dd,J=9.5,3.0 Hz,1H,H-3′′),3.90-3.81(m,4H,H-5′′,H-3,H-6 a ",H-5 a ′′′′),3.74(dd,J=10.2,8.1Hz,1H,H-2′),3.71-3.60(m,5H,H-4′′,H-6 b ′′′,H-5′′′′′,H-5′,H-3′),3.55(d,J=2.6 Hz,1H,H-4′),3.49-3.41(m,2H,H-4′′′′,H-4′′′′′′),3.33-3.25(m,7H,H-2′′′,H-3′′′,H-4′′′,H-5′′′,H-3′′′′,H-3′′′′′′,-NHC H a- ), 3.23 - 3.08 (m, 4H, H - 2′′′′, H - 5 b ′′′′, H - 2′′′′′), 2.95 - 2.88 (m, 1H, H - 18), 2.60 (t, J = 7.6 Hz, 2H, carbon chain C H 2 Ph), 2.28 (t, J = 13.6 Hz, 1H, H - 19 a ), 1.98 - 1.86 (m, 5H, H - 2 a , H - 21 a , H - 22 a , H - 11 ab ), 1.86 - 1.77 (m, 2H, H - 22 b , H - 2 b ), 1.76 - 1.67 (m, 3H, H - 9, H - 15 a , H - 1 a ), 1.65 - 1.59 (m, 2H, carbon chain C H 2 ), 1.58 - 1.48 (m, 4H, H - 6 a , H - 7 a , carbon chain C H 2 ), 1.41 - 1.38 (m, 4H, H - 15 b , H - 27), 1.37 - 1.31 (m, 9H, H - 5, carbon chain C H 2 ×4), 1.30 - 1.26 (m, 4H, H - 6 b , H - 6′′), 1.21 (d, J = 6.3 Hz, 3H, H - 6′), 1.16 (d, J = 11.2 Hz, 1H, H - 21 b ), 1.14 (s, 3H, H - 24), 1.10 (d, J = 16.7 Hz, 2H, H - 1 b ), 1.04 (d, J = 10.1 Hz, 1H, H - 19 b ), 1.00 (s, 3H, H - 25), 0.96 - 0.94 (m, 1H, H - 7 b ), 0.93 (s, 3H, H - 30), 0.85 (s, 3H, H - 29), 0.79 (s, 3H, H - 26); 13C NMR(151 MHz,MeOD)δ 209.4(C-23),177.2(C-28),171.5,159.2,156.5,144.8(C-13),139.2,130.8,130.7,124.0,123.1(C-12),120.0,119 .4,105.4(C-1′′′),105.1(C-1′′′′′),105.0(C-1′′′′),101.5(C-1′′),95.4(C-1′),83.5(C-3),83.0(C-3′′),78.7( C-4′′),78.6(C-3′′′),78.3(C-3′′′′),77.8(C-2′′′),77.6(C-3′′′′′),76.6(C-5′′′′′),75.9(C-3′),75.7(C-2′′′ ′),75.3(C-4′′′),74.9(C-2′),74.8(C-16),74.7(C-2′′′′′),73.5(C-4′),73.2(C-4′′′′′),72.7(C-5′),71.5(C-4′ ′′′),71.3(C-2′′),71.1(C-5′′′),69.1(C-5′′),67.0(C-5′′′′),62.4(C-6′′′),56.1(C-4),49.9(C-17),49.5(C-5) ,48.1(C-19),48.0(C-9),42.8(C-14),42.4(C-18),41.1(C-8),40.0,39.4(C-1),37.1(C-10),36.5(C-15,C-21),36. 2,33.8(C-6),33.4(C-29),32.9,31.8(C-22),31.3(C-20),30.7,30.5,30.4,30.3,27.9,27.3(C-27),26.0(C-2),24. 9(C-30),24.5(C-11),21.4(C-7),18.6(C-6′′),17.8(C-26),16.5(C-6′),16.4(C-25),10.6(C-24);HRMS(ESI-TOF)C 79 H 118 NO 28 [M+H] + The theoretical value is 1528.7835 and the measured value is 1528.7847.

[0158] Synthesis Example II

[0159] Coupling of trisaccharides with prosapogenins Considering the long route to conjugate glucuronic acid ester and quinoline acid ester, a semisynthetic approach was further applied to achieve the saponin core. The starting material Quillaja Ultra Dry 100-Q (Desert King, batch: QDU-100-121213-2) was treated under basic conditions to hydrolyze the C-28 linked oligosaccharides. Following triethylsilylation and selective benzylation, the prosapogenin core was obtained in three steps (Scheme 7). [ka] (Scheme 7)

[0160] Scheme 7: Isolation and selective protection of branched trisaccharide-triterpene saponins

[0161] Amide bond formation of diverse linkers on prosapogenin

[0162] Coupling of the trisaccharide with the prosapogenin can be easily accomplished to give the saponin core. Following deprotection and amide bond formation, the target saponins with diverse carbon chains were provided. [ka] (Scheme 8)

[0163] Scheme 8: Shows the preparation of saponin analogs according to embodiments of the present invention.

[0164] Experimental details [ka] 3-O-(2,3,4,6-tetra-O-triethylsilyl-β-D-galactopyranosyl-(1→2)-(2,3,4-tri-O-triethylsilyl-β-D-xylopyranosyl-(1→3))-3-O-triethylsilyl-β-D-glucopyranosyluronic acid))-16-O-triethylsilylchiric acid (81).

[0165] To a stirred suspension of prosapogenin (1.72 g) in anhydrous pyridine (25 mL), TESOTf (5.0 mL, 22.1 mmol) was added at room temperature with N 2 The reaction mixture was stirred for 2 days and TESOTf (1.3 mL, 5.8 mmol) was added, followed by one more addition after 24 h (1.0 mL, 4.4 mmol). The reaction mixture was stirred for 5 days in total. The resulting mixture was concentrated and passed through a short plug of silica gel eluted with hexanes / EtOAc (2:1). The eluent was concentrated and dried under reduced pressure to give a yellow oil. The resulting yellow oil was dissolved in MeOH / THF (1:1) (80 mL) and the solution was stirred at room temperature for 3 days. The reaction mixture was concentrated under reduced pressure and purified by column chromatography (silica gel, EtOAc / hexanes = 1 / 6 to 1 / 4) to give 81 (0.66 g, ~19%) as a white solid foam. R f 0.47 (EtOAc / benzene=1 / 4); 1 H NMR (600 MHz, CDCl 3 )δ 9.68(s,1H),5.35(br. s,1H,H-12),4.56(br. s,1H,H-16),4.54(d,J=7.4 Hz,1H,H-1'''),4.42(d,J=7.4 Hz,1H,H-1''),4.41(d,J=6.4 Hz,1H,H-1'),3.96-3.88(m,4H,H-4'',H-3',H-5',H-3''),3.84-3.82(m,2H,H-5 a ''',H-2'),3.77(t,J=9.2 Hz,1H,H-6 a ''),3.65-3.61(m,3H,H-3,H-2'',H-6 b''),3.52-3.49(m,1H,H-4'''),3.53-3.51(m,1H,H-4'),3.42-3.35(m,2H,H-3''',H-5''),3.27(t,J=7.8 Hz,1H,H-2'''),3.12(t,J=10.7 Hz,1H,H-5'''),2.96(dd,J=13.3 Hz,J=3.1Hz,1H,H-18),2.22(t,J=13.8 Hz,1H,H-19),1.92-1.86(m,4H),1.84-1.71(m,4H),1.68(t,J=8.9 Hz,1H,H-19 a ),1.63-1.31(m,1H,H-1),1.57-1.50(m,1H),1.49-1.41(m,2H,H-6),1.39-1.36(m ,5H,H-5,H-27),1.29-1.25(m,5H,H-15,H-24),1.12-1.15(m,2H,H-21), 1.11-1.07(m,1H,H-9),1.04-0.94(m,94H),0.91(s,3H,H-29),0.75-0.62 (m,54H)ppm; HRMS(ESI-TOF)C 101 H 199 O 20 S 9 [M+H] + The theoretical value was 1986.2504 and the measured value was 1986.3361. [ka] 3-O-(benzyl 2,3,4,6-tetra-O-triethylsilyl-β-D-galactopyranosyl-(1→2)-(2,3,4-tri-O-triethylsilyl-β-D-xylopyranosyl-(1→3))-3-O-triethylsilyl-β-D-glucopyranosyluronate))-16-O-triethylsilylchiric acid (82).

[0166] CH of 81 (253 mg, 127 μmol), TBP (319 mg, 1.29 mmol) and anhydrous pyridine (94 μL, 1.2 mmol). 2 Cl 2 To a stirred suspension of CBzCl (47 μL, 0.33 mmol) in N 2The reaction was added under atmospheric pressure. After 14 hours, when the reaction was complete, the reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=1 / 20-1 / 10) to give 82 (207 mg, 65%) as a white solid foam. f 0.74 (EtOAc / benzene=1 / 9); 1 H NMR (400 MHz, CDCl 3 )δ 9.68(s,1H),7.33-7.29(m,5H),5.31(br. s,1H,H-12),5.23(d,J=12.4 Hz,1H Bn C H 2 ), 5.07(d, J=12.0 Hz, 1H Bn C H 2 ),4.53(d,J=7.6 Hz,1H,H-1'''),4.51(br. s,1H,H-16),4.40(d,J=7.2 Hz,1H,H-1''),4.12(d,J=7.2 Hz,1H,H-1'),3.93-3.79(m,4H,H-4'',H-3',H-5',H-3''),3.878-3.74(m,2H,H-5 a ''',H-2'),3.72(t,J=9.1Hz,1H,H-6 a ''),3.61-3.52(m,3H,H-3,H-2'',H-6 b ''),3.49-3.42(m,1H,H-4'''),3.40-3.35(m,1H,H-4'),3.39-3.29(m,2H,H-3''',H-5''),3.23(t,J=7.7 Hz,1H,H-2'''),3.11(t,J=11.0 Hz,1H,H-5 b'''),2.91(dd,J=13.8 Hz,J=3.6 Hz,1H,H-18),2.19(t,J=13.6 Hz,1H,H-19),1.89-1.79(m,4H),1.55-1.45(m,4H),1.42-1.30(m ,5H,H-5,H-27),1.30-1.23(m,5H,H-15,H-24),1.16-1.09(m,2H,H-21),1.08-1 .01(m,1H,H-9),1.00-0.90(m,94H),0.88(s,3H,H-29),0.73-0.57(m,54H)ppm; HRMS(ESI-TOF)C 108 H 204 O 20 S 9 Na [M+Na] + The theoretical value was 2099.2803 and the measured value was 2099.3005. [ka] 3-O-(N-(8-(4-phenoxyphenyl)octyl)-β-D-galactopyranosyl-(1→2)-(β-D-xylopyranosyl-(1→3))-β-D-glucopyranosyluramido)-quilacic acid (83). To a stirred suspension of saponin diacid (26 mg, 13 μmol) and HBTU (7 mg, 25 μmol) in anhydrous THF (1 mL) was added DIPEA (5 μL, 25 μmol) and 8-(4-phenoxyphenyl)octan-1-amine (4 mg, 14 μmol) sequentially in N 2 The reaction was completed after 1 h and the reaction mixture was concentrated under reduced pressure. 2 Cl 2 Dilute with H 2 Wash twice with HO, then with brine and MgSO. 4 The mixture was dried over 1000 ml of THF and concentrated under reduced pressure. The residue was purified by flash column (silica gel, EtOAc / hexane = 1 / 20 to 1 / 10). The crude product was dissolved in 1 mL of THF and stirred for 6 h under acidic conditions at pH 1. NaHCO 3After neutralization with , the mixture was filtered through a 0.22 μmm filter plate, and the filtrate was concentrated and purified by HPLC to give product 83 (2 mg) as a white solid in 80% yield (HPLC column: SUPELCO Ascentis C18 25 cm × 10 mm, 5 μm; mobile phase: 20% ACN / H 2 O gradient ~ 90%ACN / H 2 O for 20 min, then 90% ACN / H 2 O isocratic for 15 min; flow rate: 4 mL / min): 1 H NMR (600 MHz, CD 3 OD)δ 9.44(s,1H,H-23),7.32(t,J=7.7 Hz,2H),7.16(d,J=8.3 Hz,2H),7.07(t,J=7.2 Hz,1H),6.95(d,J=8.2 Hz,2H),6.90(d,J=8.2 Hz,1H),5.28(br.s,1H,H-12),4.79(d,J=7.1Hz,1H,H-1''),4.45(s,1H,H-16),4.57(d,J=7.7 Hz,1H,H-1'''),4.43(d,J=7.4 Hz,1H,H-1'),3.89(dd,J=11.5 & 5.5 Hz,1H,H-3),3.85(dd,J=11.8 & 4.4 Hz,1H,H-5 a '''),3.81(d,J=2.5 Hz,1H,H-4'')3.75(d,J=6.2 Hz,2H,H-6''),3.70-3,63(m,4H,H-2''',H-2',H-5',H-3'),3.56-3.41(m,5H,H-2'',H-3',H-4',H-5'',H-4'''),3.26-3.19(m,4H,H-3''',H-5 b ''',-NHC H 2 -),3.00(dd,J=14.1Hz & 4.1Hz,1H,H-18),2.60(t,J=7.6 Hz,2H,carbon chain C H 2 Ph), 2.29(t, J=13.4 Hz, 1H, H-19 a),1.99-1.87(m,5H),1.80-1.72(m,3H),1.71-1.66(m,1H),1.65-1.60(m,2H),1.5 5-1.49(m,3H),1.38(s,1H,H-27),1.36-1.29(m,14H),1.15(s,3H,H-23),1.02(m= HRMS + (ESI-TOF)C 67 H 97 NO 20 Na [M+Na] + The theoretical value was 1258.6496 and the measured value was 1258.6510. [ka] 3-O-(benzyl 2,3,4,6-tetra-O-triethylsilyl-β-D-galactopyranosyl-(1→2)-(2,3,4-tri-O-triethylsilyl-β-D-xylopyranosyl-(1→3))-(3-O-triethylsilyl-β-D-glucopyranosyluronate))-28-O-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamneuranosyl-(1→2)-3,4-O-isopropylidene-β-D-fucopyranosyl)-16-O-triethylsilyl chiral ester (84).

[0167] 10 (68.8 mg, 83.6 μmol), 82 (130 mg, 62.3 μmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in (3.0 mL) 3 ·OEt 2 (approximately 48%, 4 μL, 24 μmol) at -75 °C with N 2 The reaction was complete after 0.5 h when Et 3 The reaction was quenched with N and allowed to warm to room temperature. The resulting mixture was diluted with CH 2 Cl 2The mixture was diluted with 5 μm filter paper and filtered through a 5 μm filter paper. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / CH 2 Cl 2 / hexane = 1 / 1 / 6 to 1 / 1 / 4) to give 84 (160 mg, 93%) as a white solid foam: R f 0.63 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl 3 )δ 9.67(s,1H),7.32-7.29(m,5H),5.38(d,J=7.4 Hz,1H,H-1''''),5.30(t,J=3.7 1H,H-12),5.26(d,J=12.0 Hz,1H Bn C H 2 ),5.23-5.21(m,1H,H-2'''''),5.19(dd,J=9.8 Hz,J=3.5 Hz,1H,H-3'''''),5.07(d,J=12.4 Hz,1H Bn C H 2 ),4.97(d,J=0.8 Hz,1H,H-1'''''),4.93(dt,J=9.2 Hz,J=5.5 Hz,1H,H-4''''''),4.83(dd,J=9.2 Hz,J=5.5 Hz,1H,H-2''''''),4.61(d,J=7.8 Hz,1H,H-1''''''),4.53(d,J=7.8 Hz,1H,H-1'''),4.46(s,1H,H-16),4.39(d,J=7.2 Hz,1H,H-1''),4.17-4.14(m,2H,H-1',H-3''''),4.12-4.07(m,2H,H-3',H-5 a ''''''),3.99(dd,J=5.8 Hz,J=1.9 Hz,1H,H-4''''),3.91-3.88(m,2H,H-4',H-5'),3.86-3.81(m,3H,H-4'',H-5'''',H-3''),3.81-3.75(m,3H,H-5''',H-5 a ''',H-2'),3.72(t,J=9.2 Hz,1H,H-6 a ''),3.66-3.60(m,2H,H-2'''',H-5'''''),3.59-3.53(m,3H,H-2'',H-6b '',H-3),3.47-3.42(m,1H,H-4'''),3.36(dd,J=9.4 Hz,J=2.2 Hz,1H,H-4''''),3.34-3.29(m,3H,H-3''',H-5'',H-5 b ''''''),3.22(t,J=7.4 Hz,1H,H-2'''),3.10,(t,J=11.0 Hz,1H,H-5 b '''),2.90(dd,J=14.1Hz,J=3.7 Hz,1H,H-18),2.21(t,J=13.7 Hz,1H,H-19),2.11(s,3H),2.04(s,3H),2.01(s,3H),1.99(s,3H),1.95(s,3H),1.83-1.77(m,4H,H-11,H-22),1. 76-1.62(m,4H),1.62-1.55(m,3H),1.52(s,3H,H-27),1.50-1.47(m,1H),1.33(s,1H,H-5),1.31(s,6H,isopropylidene C H 3 ),1.27(s,3H,H-24),1.26-1.25(m,3H,H-6''''),1.25-1.24(d,3H,H-6'''''),1.24-1. 22(m,4H),0.98-0.91(m,94H),0.91-0.89(m,10H,H-1,H-7,H-15,H-20),0.69-0.56(m,54 H)ppm; HRMS(ESI-TOF)C 138 H 246 O 37 S 9 Na [M+Na] + The theoretical value was 2772.5224 and the measured value was 2772.5586. [ka]

[0168] 3-O-{β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-[N-(8-(4-methoxyphenyl)octyl)-β-D-glucuropyranosyluramide]}-28-O-[β-D-xylopyranosyl-(1→4)-α-L-rhamneuranosyl-(1→2)-β-D-fucopyranosyl] chiral acid ester (92). To a stirred suspension of 50 (8 mg, 6 μmol), 8-(4-methoxyphenyl)octan-1-amine (13 mg, 58 μmol) and HBTU (22 mg, 58 μmol) in anhydrous DMA (0.5 mL) was added DIPEA (10 μL, 58 μmol) and N 2 The reaction was added under atmospheric pressure. When the reaction was complete after 24 h, the reaction mixture was concentrated under reduced pressure, diluted with MeOH, and filtered through a 5 μm filter paper. The filtrate was concentrated and purified by HPLC to give the product 51b (2 mg) as a white solid in 30% yield: (HPLC column: SUPELCO Ascentis C18 25 cm×10 mm, 5 μm; mobile phase: 30% ACN / H 2 O gradient ~ 80%ACN / H 2 O for 20 min, then 90% ACN / H 2 O isocratic for 15 min; flow rate: 5 mL / min); HRMS (ESI-TOF) C 79 H 123 NO 32 Na [M+Na] + The theoretical value was 1620.7920 and the measured value was 1620.7920. [ka] 3-O-{β-D-galactopyranosyl-(1→2)-[β-D-xylopyranosyl-(1→3)]-[N-(8-(4-phenoxyphenyl)octyl)-β-D-glucuropyranosyluramide]}-28-O-[β-D-xylopyranosyl-(1→4)-α-L-rhamneuranosyl-(1→2)-β-D-fucopyranosyl] chiral acid ester (95). To a stirred suspension of 50 (5 mg, 4 μmol), 8-(4-phenoxyphenyl)octan-1-amine (10 mg, 40 μmol) and HBTU (12 mg, 40 μmol) in anhydrous DMA (0.5 mL) was added DIPEA (6 μL, 40 μmol) and N 2 The reaction was added under atmospheric pressure. When the reaction was complete after 24 h, the reaction mixture was concentrated under reduced pressure, diluted with MeOH, and filtered through a 5 μm filter paper. The filtrate was concentrated and purified by HPLC to give the product 51a (2 mg) as a white solid in 80% yield (HPLC column: SUPELCO Ascentis C18 25 cm×10 mm, 5 μm; mobile phase: 20% ACN / H 2 O gradient ~ 90%ACN / H 2 O for 20 min, then 90% ACN / H 2 O isocratic for 15 min; flow rate: 2.4 mL / min): 1 H NMR (600 MHz, CD 3 OD) δ 9.45(s,1H,H-23), 8.54(s,1H,amide N H ),7.33(dd,J=8.5 Hz &7.6 Hz,2H),7.18(d,J=8.5 Hz,2H),7.08(t,J=7.4 Hz,1H),6.95(d,J=7.8 Hz,2H),6.91(d,J=8.5 Hz,1H),5.37(d,J=1.5 Hz,1H,H-1'''''),5.28(t,J=3.4 Hz,1H,H-12),5.27(d,J=8.2 Hz,1H,H-1''''),4.79(d,J=7.1Hz,1H,H-1''),4.62(s,1H,H-16),4.57(d,J=7.7 Hz,1H,H-1'''),4.47(d,J=7.7 Hz,1H,H-1''''''),4.43(d,J=7.4 Hz,1H,H-1'),3.93(m,1H,H-2'''''),3.89(m,1H,H-5 a '''),3.82(m,6H,H-4'',H-2'''',H-3'''',H-3''''',H-5''''',H-5 a''''''),3.75(m,2H,H-6''),3.69(m,1H,H-2'''),3.65(m,5H,H-2',H-5',H-3'',H-4'''',H-5''''),3.49(m,9 H,H-3,H-3',H-4',H-5'',H-4''',H-3''''',H-4''''',H-2'''''',H-4''''''),3.22(m,6H,H-2'',H-3''',H-5 b ''',H-3''',H-5 b -NHC H a -), 2.95(m, 2H, H-18, -NHC H b -),2.60(t,J=7.6 Hz,2H,carbon chain C H 2 Ph), 2.29(t, J=13.4 Hz, 1H, H-19 a ),1.95(m,2H),1.90(m,4H),1.76(m,3H),1.69(m,2H),1.63(m,4H),1.53(m,3 H),1.46(m,3H),1.38(s,1H,H-27),1.34(m,13H),1.30(m,11H),1.20(d,J=6.4 Hz,3H,H-6'''''),1.16(s,3H,H-23),1.15(m,1H),1.08(m,3H),0.98(s,3H, H-25),0.92(s,3H,H-30),0.90(m,2H),0.86(s,3H,H-24),0.74(s,3H,H-26); HRMS(ESI-TOF)C 84 H 125 NO 32 Na [M+Na] + The theoretical value was 1682.8077 and the measured value was 1682.8079. 6-N-Glycosyl chelate

[0169] Coupling of the chelate with azido-glucose was successfully achieved to give product 117 in 70% yield. Interestingly, the results revealed a preference for 3-O glycosylation over the 16-O position. [ka]

[0170] With glycoside 117 in hand, further modifications were carried out to unmask the C-28 carboxylic acid. First, the benzoyl group was hydrolyzed at elevated temperature under basic conditions. Surprisingly, the 28-O-allyl ester was unaffected under this harsh environment. After triethylsilylation of the resulting azido glycoside, the azide group was reduced to an amine and coupled with a lipophilic long-chain acid to form an amide, affording the fully protected chiral acid esters 119a / b / c. The O-allyl esters 119a / b / c were then converted to chiral esters 119b / c by the addition of Pd(OAc) under mildly acidic conditions. 2 The monoacids 120a / b / c were coupled with the trisaccharide 10 under Lewis acid catalysis at -78 °C to give 121a / b / c in 56% yield. The fully protected saponins 121a / b / c were then hydrolyzed by H 2 Pd(OAc) in THF / MeOH under atmosphere 2 The benzyl groups on the 3-O and 4-O of glucose were hydrolyzed by suspending the 122a / b / c in 13%, 16%, and 30% yields after acidic hydrolysis and basic methanolysis. [ka] 3-O-(6-azido-2-O-benzoyl-3,4-di-O-benzyl-6-deoxy-β-D-glucopyranosyl)-28-O-allyl-chiral acid ester (117): 1 H NMR (600 MHz, CDCl 3 )δ 9.17(s,1H,H-23),8.04-8.01(m,2H,Bz),7.61-7.56(m,1H,Bz),7.48-7.44( m,2H,Bz),7.36-7.24(m,5H,Bn),7.14-7.09(m,5H,Bn),5.88-5.80(m,1H,all internal alkenyl C H ), 5.33(t,J=3.5 Hz,1H,H-12), 5.28(d,J=17.6 Hz,1H,all terminal alkenyl C Ha ),5.21-15(m,2H,H-2′,Full end アルケニルC H b ),4.85(d,J=11.2 Hz,1H,Bn C H a ),4.68(d,J=11.1Hz,1H,Bn C H b ),4.62(d,J=11.1Hz,1H,Bn C H a ),4.57(d,J=11.2 Hz,1H,Bn C H b ),4.52-4.45(m,3H,H-16,アリルic C H 2 ),4.43(d,J=7.9 Hz 1H,H-1′),3.80-3.73(m,2H,H-3,H-3′),3.53-3.50(m,2H,H-4′,H-5′),3.41-3.38(m,1H,H-6 a ′), 3.32-3.28(m,1H,H-6 b ′),3.05(dd,J=14.4,4.4 Hz,1H,H-18),2.13(t,J=13.4 Hz,1H,H-19),1.93-1.82(m,4H),1.80-1.69(m,4H),1.66-1.60(m,3H),1.59(br. s,3H),1.43-1.33(m,2H),1.31(s,3H),1.30-1.26(m,1H),1.21-1.14(m,3H),1.10(dd,J=12.8,3.6 Hz,1H),1.03-0.99(m,1H),0.97-0.94(m,6H),0.90-0.88(m,6H)ppm; HRMS(ESI-TOF)C 60 H 75 N 3 O 10 Na [M+Na] + The theoretical value is 1020.5345 and the measured value is 1020.5350.

change

[0171] 3-O-(3,4-di-O-benzyl-6-deoxy-6-(11-(4-(4-fluorophenoxyl)phenyl)undecanamido)-2-O-triethylsilyl-β-D-glucopyranosyl)-16-O-triethylsilyl-28-O-allyl-chloyl ester (119a)

[0172] To a stirred solution of 118 (282 mg, 0.25 mmol) in THF (15 mL) was added PPh 3 (200 mg, 0.76 mmol) was added. The mixture was stirred for 12 h, then 0.5 mL of H 2 O was added and the THF was removed under reduced pressure at 35 °C. The resulting residue was 2 Cl 2 Dilute with H 2 o, washed with brine, and MgSO 4 The mixture was dried over 500 ml and concentrated under reduced pressure. A stirred solution of the resulting mixture in THF (7 mL) was treated with a premixed suspension of 11-(4-(4-fluorophenoxy)phenyl)undecanoic acid (187 mg, 0.50 mmol), HBTU (286 mg, 0.75 mmol), DIPEA (132 μL, 0.75 mmol) and THF (7 mL). After stirring at 30° C. for 2 h, the reaction was complete and the residue was concentrated under reduced pressure to remove THF. The residue was purified by hexane distillation (HCl), 100%. 2 Cl 2 Dilute with H 2 o, washed with brine, and MgSO 4 The mixture was dried at 40° C. and concentrated under pressure. The residue was purified by column chromatography (silica gel, EtOAc / hexane=1 / 8) to give 119a (349 mg, 96%) as a white solid foam: R f 0.29 (EtOAc / Hexane=1 / 5); 1 H NMR (600 MHz, CDCl 3)δ 9.45(s,1H,H-23),7.33-7.26(m,4H),7.22-7.17(m,2H),7.10(d,J = 8.5 Hz,2H),7.01-6.96(m,2H),6.95-6.91(m,2H),6.86(d,J = 8.5 Hz,2H),5.87-5.81(m,1H,internal alkenyl C H ),5.32(t,J = 3.7 Hz,1H,H-12),5.28(dq,J = 17.0 Hz & 1.2 Hz,1H,terminal alkenyl C H a ),5.18(dt,J = 10.6 Hz & 1.2 Hz,1H,terminal alkenyl C H b ),4.83(q,J = 10.5 Hz,2H,Bn C H 2 ),4.67(d,J = 10.5 Hz,1H,Bn C H a ),4.56(br. s,1H,H-16),4.51(d,J = 10.6 Hz,1H,Bn C H b ),4.45(dt,J = 19.9 Hz & 1.4 Hz,2H,allylic C H 2 ),4.02(d,J = 6.8 Hz,1H,H-1′),3.93(dd,J = 11.2 Hz & 5.2 Hz,1H,H-3),3.59(dt,J = 13.9 Hz & 5.9 Hz,1H,H-6 a ′),3.47(dt,J = 13.9 Hz & 5.9 Hz,1H,H-6 b ′),3.43-3.37(m,2H,H-2′,H-4′),3.36-3.22(m,2H,H-3′,H-5′),3.00(dd,J = 14.3 Hz & 4.0 Hz,1H,H-18),2.54(t,J = 6.7 Hz,2H,C H 2 PhOPhF),2.32(t,J = 7.6 Hz,1H,NHC H a ),2.20(m,1H,H-19 a ),2.15(td,J = 13.6 Hz & 3.1Hz,1H,H-19 b),1.90-1.85(m,2H),1.84-1.76(m,3H),1.72-1.65(m,3H),1.64-1.53(m,9H),1.48-1.40(m,2H),1.34(s,2H),1.31-1.23(m,22H) ,1.16(s,2H),1.13-1.08(m,2H),1.06-1.02(m,2H),1.00-0.95(m,10H),0.95-0.92(m,4H),0.92-0.91(m,2H),0.91-0.90(m,3H), 0.89-0.88(m,2H),0.87-0.85(m,4H),0.69-0.63(m,8H),0.63-0.57(m,6H)ppm; HRMS + (ESI-TOF)C 88 H 129 FNO 11 S 2 [M+Na] + The theoretical value was 1451.9113 and the measured value was 1451.9095. [ka]

[0173] 3-O-(3,4-di-O-benzyl-6-decanoic acid-6-deoxy-2-O-triethylsilyl-β-D-glucopyranosyl)-16-O-triethylsilyl-28-O-allyl-chloate (119b)

[0174] Following the azide reduction and amide formation procedure as described above, 119b was obtained as a white solid in 56% yield: 1 H NMR (600 MHz, CDCl 3 ) δ 9.40(s,1H,H-23),7.33-7.24(m,5H),7.24-7.15(m,5H),5.87-5.80(m,1H,internal alkenyl C H ), 5.31 (t, J = 3.4 Hz, 1H, H-12), 5.30-5.25 (m, 2H, terminal alkenyl C H a ,N H CH 2 ), 5.18(dd, J=10.5 Hz & 1.1Hz, 1H, terminal alkenyl CH b ),4.87-4.80(m,2H,Bn C H 2 ),4.65(d,J=10.4 Hz,1H,Bn C H a ),4.58-4.52(m,2H,H-16,Bn C H b ),4.46(qd,J=13.6 Hz & 1.1Hz,2H,アリルic C H 2 ),4.07(d,J=6.8 Hz,1H,H-1′),3.98-3.91(m,1H,H-3),3.65-3.54(m,1H,H-4′),3.50-3.30(m,4H,H-2′,H-5′,H-6′),3.29-3.22(m,1H,H-3′),3.00(dd,J=14.2 Hz & 3.8 Hz,1H,H-18),2.31(t,J=7.4 Hz,1H,NHC H a ),2.20(t,J=12.8 Hz,1H,H-19 a ),1.90-1.75(m,7H),1.72-1.63(m,4H),1.63-1.58(m,3H),1.45-1.40(m,2H),1.39(s,3 H),1.31-1.21(m,16H),1.78-1.15(m,1H),1.12(s,3H),1.06-1.02(m,2H),0.98(t,J=3.7 Hz,3H),0.97(s,3H),0.96(br. s,2H),0.95-0.92(m,6H),0.91(br. HRMS + (ESI-TOF)C 75 H 120 NO 10 Si 2 [M+H] + The theoretical value is 1251.8475 and the measured value is 1251.8426.

change

[0175] 3-O-{2-O-triethylsilyl-3,4-di-O-benzyl-6-[9-(4-phenoxyl-phenyl)nonanamide]-6-deoxy-β-D-glucopyranosyl}-16-O-triethylsilyl-28-O-allyl-chloate (119c)

[0176] Following the azide reduction and amide formation procedure as described above, 119b was obtained as a white solid in 56% yield: 1 H NMR (400 MHz, CDCl 3 )δ 9.45(s,1H,H-23),7.31-7.21(m,2H),7.10(d,J=8.4 Hz,2H),7.04(t,J=7.4 Hz,2H),6.96(dd,J=8.7 Hz & 1.0 Hz,2H),6.90(d,J=8.5 Hz,2H),5.89-5.80(m,2H,Amide N H ,internal alkenyl C H ), 5.32(t,J=3.7 Hz,1H,H-12), 5.28(d,J=18.2 Hz,1H,terminal alkenyl C H a ), 5.17(dd, J = 10.4 Hz & 1.3 Hz, 1H, terminal alkenyl C H b ), 4.83(q,J=11.7 Hz,2H,Bn C H 2 ), 4.67(d, J=10.4 Hz, 1H, Bn C H a ),4.57(br. s,1H,H-16),4.51(d,J=10.4 Hz,1H,Bn C H b ), 4.45(ddt,J=11.4 Hz & 5.6 Hz & 1.1Hz,2H,allylic C H 2),4.02(d,J=6.8 Hz,1H,H-1′),3.94(dd,J=10.2 Hz & 5.8 Hz,1H,H-3),3.63-3.56(m,1H),3.51-3.39(m,3H),3.32- 3.21(m,2H),3.00(dd,J=14.2 Hz & 3.9 Hz,1H,H-18),2.55(t,J=7.5 Hz,2H,C H 2 PhOPh), 2.33-2.13(m,3H,H-19 a ,NHCOC H 2 ),1.91-1.83(m,3H),1.83-1.76(m,3H),1.75-1.52(m,11H),1.49-1.40(m,2H),1.34(s,3H),1.29(br. BBD 13 C NMR (100 MHz, CDCl 3 )δ 207.5(C-23),176.4(C-28),173.1(NH C O),157.7,154.8,143.5(C-13),138.5,137.8,137.5,132.2,129.6 129.4,128.4,128.2,128.1,127.9,127.2,126.8,122.7,121.7(C-12),118.9,118.4,117.8(Full end アルケニル C H 2 ),101.0(C-1′),85.5(C-3),79.3,79.0,75.2(Bn C H 2 ),75.0,75.0(Bn C H 2),74.9(C-16),73.1,65.0(C-6′),54.5,48.9,48.8,46.6,46.3,41.3,40.4,40.0,39.5,38.1,36.7,36.0,35.2,35.1,34.5,33.8,32 .7,32.3,31.5×2,30.5,29.6,29.3×2,29.2,29.1,29.0,26.3,25.7,24.7,24.6,24.2,23.2,20.1,16.9,15.5,10.4,7.1,6.9,5.0,4.9 ppm; HRMS + (ESI-TOF)C 86 H 125 NO 11 S 2 [M+H] + The theoretical value was 140.8894 and the measured value was 1405.8984.

[0177] [ka]

[0178] 3-O-(3,4-di-O-benzyl-6-deoxy-6-(11-(4-(4-fluorophenoxyl)phenyl)undecanamido)-2-O-triethylsilyl-β-D-glucopyranosyl)-16-O-triethylsilyl-chiric acid (120a)

[0179] 119a (237 mg, 0.16 mmol) and PPh 3 To a stirred solution of (107 mg, 0.41 mmol) in 1,4-dioxane (4 mL) was added a solution of formic acid (129 μL, 3.4 mmol) in 1,4-dioxane (2 mL) premixed with Et 3 Pd(OAc) in N (456 μL, 3.2 mmol) and 1,4-dioxane (2 mL) 2 (18 mg, 0.08 mmol) was added at room temperature. The reaction mixture was stirred for 12 hours and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel: EtOAc / hexane = 1 / 4 to 1 / 2) to give 120a (186 mg, 81%) as a white solid: R f 0.36 (EtOAc / Hexane=1 / 2);1 H NMR(600 MHz,CDCl 3 )δ 9.41(s,1H,H-23),7.27-7.26(m,1H),7.25(br. s,1H),7.34-7.25(m,6H),7.06(d,J=8.4 Hz,1H),6.98-6.64(m,6H),6.82(d,J=8.4 Hz,1H),5.82(t,J=5.2 Hz,アミド N H ),5.30(br. s,1H,H-12),4.83-4.76(m,2H,Bn C H 2 ),4.63(d,J=10.3 Hz,1H,Bn C H a ),4.50-4.53(m,2H,H-16,Bn C H b ),3.98(d,J=6.6 Hz,1H,H-1′),3.90(dd,J=11.2 Hz & 4.9 Hz,1H,H-3),3.60-3.53(m,1H,H-6 a ′),3.48-3.41(m,1H,H-6 b ′),3.41-3.34(m,2H,H-2′,H-4′),3.27-3.19(m,2H,H-3′,H-5′),2.90(dd,J=14.1Hz & 3.6 Hz,1H,H-18),2.50(t,J=7.6 Hz,2H,C H 2 PhOPhF),2.31(t,J=7.4 Hz,1H,NHC H a ),2.19-2.09(m,3H),1.87-1.68(m,7H),1.68-1.59(m,3H),1.59-1.49(m,4H),1.44-1.36(m,2H),1.30(s,3H,H-27),1.28-1.16(m,17H),1.11(s,3H,H-29),1.09-1.08(m,1H),1.03-0.97(m,2H),0.94(s,2H),0.93(s,3H),0.92(s,3H),0.89-0.87(m,5H),0.87(s,3H),0.85(s,2H),0.82(s,3H),0.66-0.53(m,17H)ppm; HRMS + (ESI-TOF)C 85 H125 FNO 11 S 2 [M+H] + The theoretical value was 1411.8800 and the measured value was 1411.8742. [ka]

[0180] 3-O-(2-O-triethylsilyl-3,4-di-O-benzyl-6-decanoic acid-6-deoxy-β-D-glucopyranosyl)-16-O-triethylsilyl-28-O-allyl-quinoline acid (120b)

[0181] 119b (200 mg, 0.16 mmol) and PPh 3 To a stirred solution of (107 mg, 0.41 mmol) in 1,4-dioxane (4 mL) was added a solution of formic acid (129 μL, 3.4 mmol) in 1,4-dioxane (2 mL) premixed with Et 3 Pd(OAc) in N (456 μL, 3.2 mmol) and 1,4-dioxane (2 mL) 2 (18 mg, 0.08 mmol) was added at room temperature. The reaction mixture was stirred for 12 h and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, EtOAc / hexane=1 / 4-1 / 2) to give 120b (154 mg, 80%) as a white solid: R f 0.36 (EtOAc / Hexane=1 / 2); 1 H NMR (600 MHz, CDCl 3 ) δ 9.45(s,1H,H-23), 7.32-7.24(m,6H), 7.24-7.19(m,4H), 5.85(t,J=5.0 Hz, amide N H ),5.30(t,J=3.7 Hz 1H,H-12),4.86-4.80(m,2H,Bn C H 2 ), 4.66(d, J=10.4 Hz, 1H, Bn C H a ), 4.54-4.49(m,2H,H-16,Bn C H b),4.03(d,J=6.8 Hz,1H,H-1′),3.94(dd,J=11.2 Hz & 4.9 Hz,1H,H-3),3.59-3.55(m,1H),3.52-3.46(m,1H),3.44-3.38(m,2H),3.31-3.24(m,2H),2.94(dd,J=14.2 Hz & 4.0 Hz,1H,H-18),2.31(t,J=7.5 Hz,1H,NHC H a ),2.19-2.09(m,3H),1.87-1.68(m,7H),1.68-1.59(m,3H),1.59-1.49(m,4H),1.44-1.36(m,2H),1.30(s,3H,H-27),1.28-1.16(m,17H),1.11(s,3H),1.09-1.08(m,1H),1.03-0.97(m,2H),0.94(s,2H),0.93(s,3H),0.92(s,3H),0.89-0.87(m,5H),0.87(s,3H),0.85(s,2H),0.82(s,3H),0.66-0.53(m,17H); BBD 13 C NMR(150 MHz,CDCl 3 )δ 207.4(C-23),182.7(C-28),173.3(NH C O),143.3(C-13),138.6,137.6,128.4,128.2,126.9,121.8(C-12),101.1(C-1′),85.5,79.5,79.2(C-3),75.2(Bn C H 2 ),75.0(Bn C H 2 ),74.9,74.8(C-16),73.2,54.5,49.0,48.6,46.6,46.3,41.3,40.1(C-6′),39.5(C-8),38.2,36.8(-NHCO C H 2 -),36.1 ,35.1,34.6,34.0,32.6,32.,31.8,31.6,30.5,29.5,29.4,29.3,29.2,29.0,26.4,25.8,24.7,24.2,23.2,22.7,20.1,16.9,15.5,14.1,10.4,7.1,7.0,5.1,5.0 ppm; HRMS+ (ESI-TOF)C 72 H 116 NO 10 S 2 [M+H] + The theoretical value was 1210.8132 and the measured value was 1210.8108. [ka]

[0182] 3-O-{2-O-triethylsilyl-3,4-di-O-benzyl-6-[9-(4-phenoxyl-phenyl)nonanamide]-6-deoxy-β-D-glucopyranosyl}-16-O-triethylsilyl-28 quilacic acid (120c)

[0183] 119c (224 mg, 0.16 mmol) and PPh 3 To a stirred solution of (107 mg, 0.41 mmol) in 1,4-dioxane (4 mL) was added a solution of formic acid (129 μL, 3.4 mmol) in 1,4-dioxane (2 mL) premixed with Et 3 Pd(OAc) in N (456 μL, 3.2 mmol) and 1,4-dioxane (2 mL) 2 (18 mg, 0.08 mmol) was added at room temperature. The reaction mixture was stirred for 12 h and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, EtOAc / hexane=1 / 4-1 / 2) to give 120c (154 mg, 80%) as a white solid: R f 0.36 (EtOAc / Hexane=1 / 2) 1 H NMR (400 MHz, CDCl 3 )δ 9.45(s,1H,H-23),7.31-7.21(m,2H),7.10(d,J=8.4 Hz,2H),7.04(t,J=7.4 Hz,2H),6.96(dd,J=8.7 Hz & 1.0 Hz,2H),6.90(d,J=8.5 Hz,2H),5.89-5.80(m,2H,Amide N H ,internal alkenyl C H),5.32(t,J=3.7 Hz,1H,H-12),4.83(q,J=11.7 Hz,2H,Bn C H 2 ),4.67(d,J=10.4 Hz,1H,Bn C H a ),4.57(br. s,1H,H-16),4.51(d,J=10.4 Hz,1H,Bn C H b ),4.02(d,J=6.8 Hz,1H,H-1′),3.94(dd,J=10.2 Hz & 5.8 Hz,1H,H-3),3.63-3.56(m,1H),3.51-3.39(m,3H),3.32- 3.21(m,2H),3.00(dd,J=14.2 Hz & 3.9 Hz,1H,H-18),2.55(t,J=7.5 Hz,2H,C H 2 PhOPh),2.33-2.13(m,3H,H-19 a ,NHCOC H 2 ),1.91-1.83(m,3H),1.83-1.76(m,3H),1.75-1.52(m,11H),1.49-1.40(m,2H),1.34(s,3H),1.29(br. s,11H),1.23(s,3H),1.15(s,3H),1.14-1.00(m,4H),0.98-0.86(m,31H),0.69-0.57(m,16H); BBD 13 C NMR(100 MHz,CDCl 3 )δ 207.5(C-23),182.0(C-28),173.2(NH C O),157.7,154.8,143.5(C-13),138.5,137.5,132.2,129.6 129.4,128.4,128.2,128.1,127.9,127.2,126.8,122.7,121.7(C-12),118.9,118.4,101.0(C-1′),85.5(C-3),79.3,79.0,75.2(Bn C H 2 ),75.0,75.0(Bn C H 2),74.9(C-16),73.1,65.0(C-6′),54.5,48.9,48.8,46.6,46.3,41.3,40.4,40.0,39.5,38.1,36.7,36.0,35.2,35.1,34.5,33.8,32 .7,32.3,31.5×2,30.5,29.6,29.3×2,29.2,29.1,29.0,26.3,25.7,24.7,24.6,24.2,23.2,20.1,16.9,15.5,10.4,7.1,6.9,5.0,4.9 ppm; HRMS + (ESI-TOF)C 83 H 122 NO 11 S 2 [M+H] + The theoretical value was 1364.8551 and the measured value was 1364.8567. [ka]

[0184] 3-O-(3,4-di-O-benzyl-6-deoxy-6-(11-(4-(4-fluorophenoxyl)phenyl)undecanamido)-2-O-triethylsilyl-β-D-glucopyranosyl)-28-O-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamneuranosyl-(1→2)-3,4-O-isopropylidene-β-D-fucopyranosyl)-16-O-triethylsilyl chiral acid ester (121a)

[0185] 10 (40 mg, 48 μmol), 120a (50 mg, 35 μmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in (0.5 mL) 3 ·OEt 2 (approximately 48%, 4 μL, 24 μmol) at -75 °C with N 2 The reaction was complete after 0.5 h, and Et 3 The reaction was quenched with N and allowed to warm to room temperature. The resulting mixture was diluted with CH 2 Cl 2The mixture was diluted with 1,000 ml of ethyl acetate and filtered through a 5 μm filter paper. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=1 / 5-1 / 2) to give 121a (37 mg, 50%) as a white solid foam. f 0.56 (EtOAc / Hexane=1 / 1); 1 H NMR (600 MHz, CDCl 3 )δ 9.49(s,1H,H-23),7.33-7.26(m,6H),7.26-7.21(m,4H),7.12(d,J=8.4 Hz,1H),7.02-7.00(m,2H),6.96-6.93(m,2H),6.88(d,J=8.6 Hz,1H),6.82(d,J=8.4 Hz,1H),5.86(t,J=5.1Hz,amide N H ),5.41(d,J=7.6 Hz,1H,H-1′′),5.33(t,J=3.4 Hz,1H,H-12),5.25(dd,J=3.4 Hz & 1.3 Hz,1H,H-2′′′),5.20(dd,J=9.8 Hz & 3.5 Hz,1H,H-3′′′′),5.13(t,J=9.4 Hz,1H,H-3′′′′),4.98(d,J=1.3 Hz,1H,H-1′′′),4.97-4.94(m,1H,H-4′′′′),4.88-4.82(m,3H,H-2′′′′′,Bn C H 2 ), 5.68(d, J=10.4 Hz, 1H, Bn C H a ),4.63(d,J=7.7 Hz,1H,H-1′′′′′),4.53(d,J=10.4 Hz,1H,Bn C H b ),4.50(br. s,1H,H-16),4.17(t,J=6.0 Hz,1H,H-3′′),4.12-4.10(m,1H,H-5 a ′′′′),4.04-4.00(m,2H,H-1′,H-4′′),3.98-3.94(m,1H,H-3),3.89-3.84(m,1H,H -5′′),3.84-3.79(m,1H,H-5′′′),3.68-3,64(m,1H,H-2′′),3.64-3.59(m,2H,H-6 a′,H-4′),3.49(dt,J=13.9 Hz,& 3.8 Hz,1H H-6 b ′),3.45-3.39(m,2H,H-2′,H-4′),3.36-3.24(m,3H,H-3′,H-5′,H-5 b ′′′′),2.93(dd,J=14.2 Hz & 3.8 Hz,1H,H-18),2.56(t,J=7.6 Hz,2H,C H 2 PhOPhF),2.22(m,1H,H-19 a ),2.17(td,J=7.5 Hz & 3.4 Hz,2H,-NHCOC H 2 -),2.13(s,3H),2.06(s,3H),2.03(s,3H),2.01(s,3H),1.98(s,3H),1.90-1.86(m,2H),1.86-1.82(m,1H),1.83-1.76(m,3H),1.74-1.67(m,4H),1.67-1.56(m,5H),1.52(s,3H,イソプロピルidne C H 3 ),1.52-1.49(m,1H),1.34(s,3H,H-27),1.33(s,3H,イソプロピリデン C H 3 ),1.30-1.25(m,24H),1.23-1.32(m,1H),1.19(s,3H,H-24),1.13-1.10(m,1H),1.05-1.03(m,1H),1.03-1.01(m,1H), 1.01-0.96(m,15H,H-25,TES CH 3 ×4),0.92(s,H,H-30),0.91(s,3H,TES CH 3 ),0.90(s,3H,TES CH 3 ),0.88(s,3H,H-29),0.74(s,3H,H-26),0.68-0.58(m,12H,TES CH 2 ×6)ppm; HRMS + (ESI-TOF)C 115 H 167 FNO 28 Si 2 [M+H] +The theoretical value was 2086.1223 and the measured value was 2086.1222. [ka]

[0186] 3-O-(3,4-di-O-benzyl-6-decanamido-6-deoxy-β-D-glucopyranosyl)-28-O-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamneuranosyl-(1→2)-3,4-O-isopropylidene-β-D-fucopyranosyl)-16-O-triethylsilyl chrylate (121b)

[0187] 10 (40 mg, 48 μmol), 120b (42 mg, 35 μmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in (0.5 mL) 3 ·OEt 2 (approximately 48%, 4 μL, 24 μmol) at -75 °C with N 2 When the reaction was complete after 0.5 h, Et 3 The reaction was quenched with N and allowed to warm to room temperature. The resulting mixture was diluted with CH 2 Cl 2 The mixture was diluted with 1,2-dichloromethane and filtered through a 5 μm filter paper. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=1 / 5-1 / 2) to give 121b (37 mg, 50%) as a white solid foam. f 0.56 (EtOAc / Hexane=1 / 1); Obtained 121bα / β mixture as a white solid foam. [ka]

[0188] 3-O-(2-O-triethylsilyl-3,4-di-O-benzyl-6-[9-(4-phenoxyl-phenyl)nonanamide}-28-O-(2,3,4-tri-O-acetyl-β-D-xylopyranosyl-(1→4)-2,3-di-O-acetyl-α-L-rhamneuranosyl-(1→2)-3,4-O-isopropylidene-β-D-fucopyranosyl)-16-O-triethylsilyl xylate (121c)

[0189] 10 (40 mg, 48 μmol), 120c (42 mg, 35 μmol) and activated 4 Å molecular sieve powder were dissolved in anhydrous CH 2 Cl 2 To a stirred suspension in (0.5 mL) 3 ·OEt 2 (approximately 48%, 4 μL, 24 μmol) at -75 °C with N 2 When the reaction was complete after 0.5 h, Et 3 The reaction was quenched with N and allowed to warm to room temperature. The resulting mixture was diluted with CH 2 Cl 2 The mixture was diluted with 5 μm filter paper and filtered through a 5 μm filter paper. The filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexane=1 / 5-1 / 2) to give 121c (37 mg, 50%) as a white solid foam. f 0.56 (EtOAc / Hexane=1 / 1); 121cα / β mixture was obtained. [ka]

[0190] 3-O-(6-deoxy-6-(11-(4-(4-fluorophenoxyl)phenyl)undecanamido)-β-D-glucopyranosyl)-28-O-(β-D-xylopyranosyl-(1→4)-α-L-rhamneuranosyl-(1→2)-β-D-fucopyranosyl) chiral acid ester (122a)

[0191] 121a (32 mg, 15 μmol) and 10% Pd(OH) 2A suspension of 5 mg / C (4 μmol) in 1.5 mL of THF / MeOH (4 / 1) was heated at 1 atm of H 2 The reaction mixture was stirred under atmospheric pressure for 12 hours. 2 Cl 2 To a stirred solution in (0.5 mL) of pre-cooled TFA / H 2 0=4 / 1 (0.5 mL) was added at 0° C. and stirred for 30 min. The solvent was evaporated under reduced pressure (<1 torr) at 0° C. and dried under high vacuum at room temperature for 1 h. A stirred solution of the residue in MeOH (1 mL) was added with K 2 CO 3 (40 mg, 300 μmol) was added and stirred for 12 h. The suspension was filtered, concentrated, and purified by HPLC to give the product 122a (3.6 mg) as a white solid in 16% yield (HPLC column: SUPELCO Ascentis C18 25 cm×10 mm, 5 μm; mobile phase: 20% ACN / H 2 O gradient ~ 90%ACN / H 2 O for 25 min, then 90% ACN / H 2 O isocratic for 15 min; flow rate: 5 mL / min): 1 H NMR (600 MHz, CD 3 OD)δ 9.41(s,1H,H-23),7.16(d,J=8.4 Hz,2H),7.07(t,J=8.8 Hz,2H),6.98-6.95(m,4H),6.87(d,J=8.6 Hz,2H),5.40(d,J=1.4 Hz,1H,H-1′′′),5.31(br. s,1H,H-12),5.29(d,J=8.2 Hz,1H,H-1′′),4.50-4.47(m,2H,H-16,H-1′′′′),4.14(d,J=7.7 Hz,1H,H-1′),3.91-3.89(m,1H,H-2′′′),3.86-3.78(m,5H,H-3,H-2′′,H-3′′′,H-5′′′,H-5 a ′′′′),3.68-3.64(m,2H,H-3′′,H-5′′),3.60-3.56(m,1H,H-6 a′),3.56-3.53(m,2H,H-4′′,H-4′′′),3.49-3.42(m,2H,H-5′,H-4′′′′),3.29-3.25(m,3H,H-3′,H-6 b ′,H-3′′′′),3.23-3.16(m,2H,H-2′′′′,H-5 b ′′′′),3.09-3.05(m,2H,H-2′,H-4′),2.45(dd,J=13.4 Hz & 3.2 Hz,H-18),2.60(t,J=7.6 Hz,2H,C H 2 PhOPhF), 2.30(m,1H,H-19 a ),2.21(t,J=7.6 Hz,2H,-NHCOC H 2 -), 1.99-1.95(m,1H), 1.95-1.91(m,1H), 1.80-1.71(m,4H), 1.70-1.65(m,1H), 1.64-1.59(m,4H), 1.55-1.50(m,2H)1.49-1.44(m,1H), 1.40(s,3H,H-27), 1.36-1.29(m,19H,H-6′′′, carbonyl C H 2 ×8),1.20(d,J=6.5 Hz,3H,H-6′′),1.12(s,3H,H-24),1.10-1.06(m,1H),1.01(s,3H,25),0.99-0.96( m,1H),0.93(s,3H,H-30),0.92-0.89(m,1H),0.87(s,3H,H-29),0.77(s,3H,H-26); BBD 13 C NMR (150 MHz, CD 3 OD)δ 209.3(C-23),177.1(C-28),176.7(アミドNH CO), 160 (d, J = 240 Hz), 156.8, 155.1, 144.9 (C-13), 139.2, 130.8, 123.1 (C-12), 121.2 (d, J = 8 Hz), 119.5, 117.2 (d, J = 23 Hz), 107.0, (C-1′′′′), 105.0 (C-1′), 101.1 (C-1′′′), 95.2 (C-1′′), 84.0 (C-4′′′), 83.4 (C-3), 78.2 (C-3′′′′), 77.6 (C-3′), 76.7 (C-3′′), 76.1 (C-2′′′′), 75.6 (C-2′), 75.2 (C-5′), 74.6 (C-16), 74.0 (C-2′′), 73.6 (C-4′′), 73.3 (C-4′), 72.7 (C-5′′), 72.2 (C-3′′′), 71.9 (C-2′′′), 71.1 (C-4′′′′), 68.7 (C-5′′′), 67.3 (C-5′′′′), 56.1 (C-5), 50.0 (C-17), 48.1 (C-19, C-9), 42.8 (C-14), 42.4 (C-18), 41.9 (C-6′), 41.1 (C-8), 39.6 (C-1), 37.3 (C-1), 36.8 (-NHCO C H 2 -), 37.1 (C-10),, 36.5 (C-21), 36.2 (- C H 2 PhOPhF), 33.6 (C-16), 3.4 (C-29), 32.9 (C-6), 32.0 (C-22), 31.4 (C-20), 30.8, 30.7, 30.6, 30.4, 27.3 (C-2), 20.2 (C-27), 26.0, 25.9 (C-2), 24.9 (C-30), 24.6 (C-11), 21.6, (C-7), 18.3 (C-6′′′), 17.7 (C-26), 16.5 (C-6′′), 16.4 (C-25), 10.6 (C-24) ppm; HRMS + (ESI-TOF)C 76 H 112 FNO 23 Si 2 Na [M + H] + The theoretical value was 1448.7501 and the measured value was 1448.7558 at

Chemical Structure

[0192] 3-O-{6-decanamido-6-deoxy-β-D-glucopyranosyl}-28-O-[β-D-xylopyranosyl-(1→4)-α-L-rhamneuranosyl-(1→2)-β-D-fucopyranosyl] chiral acid ester (122b)

[0193] 121b (28 mg, 15 μmol) and 10% Pd(OH) 2 A suspension of 5 mg / C (4 μmol) in 1.5 mL of THF / MeOH (4 / 1) was heated at 1 atm of H 2 The reaction mixture was stirred under atmospheric pressure for 12 hours. 2 Cl 2 To a stirred solution in MeOH (0.5 mL) was added pre-cooled TFA / H2O = 4 / 1 (0.5 mL) at 0 °C and stirred for 30 min. The solvent was evaporated under reduced pressure (<1 torr) at 0 °C and dried under high vacuum at room temperature for 1 h. To a stirred solution of the residue in MeOH (1 mL) was added K 2 CO 3 (40 mg, 300 μmol) was added and stirred for 12 h. The suspension was filtered, concentrated, and purified by HPLC to give the product 122b (2.3 mg) as a white solid in 13% yield (HPLC column: SUPELCO Ascentis C18 25 cm×10 mm, 5 μm; mobile phase: 20% ACN / H 2 O gradient ~ 90%ACN / H 2 O for 25 min, then 90% ACN / H 2 O isocratic for 15 min; flow rate: 5 mL / min): 1 H NMR (600 MHz, CD 3OD)δ 9.42(s,1H,H-23),5.40(s,1H,H-1′′′),5.31(br. s,1H,H-12),5.29(d,J=8.2 Hz,1H,H-1′′),4.51-4.46(m,2H,H-16,H-1′′′′),4.15(d,J=7.7 Hz,1H,H-1′),3.93-3.90(m,1H),3.87-3.78(m,5H),3.71-3.66(m,2H,),3.60-3.53(m,3H),3.49-3.41(m,2H),3.29-3.25(m,3H),3.24-3.17(m,2H),3.09-3.04(m,2H),2.95(d,J=14.1Hz,1H,H-18),2.30(m,1H,H-19a),2.21(t,J=7.6 Hz,2H,-NHCOCH2-),2.04-1.88(m,3H),1.69-1.60(m,3H),1.55-1.44(m,4H),1.40(s,3H,H-27),1.35-1.29(m,18H),1.20(d,J=6.5 Hz,3H,H-6′′),1.12(s,3H),1.10-1.06(m,1H),1.01(s,3H),0.99-0.96(m,1H),0.95(s,3H),0.93-0.89(m,3H),0.88(s,3H),0.77(s,3H); BBD 13C NMR(150 MHz,CD3OD)δ 209.4(C-23),177.2(C-28),176.7(アミド NHCO),144.9(C-13),123.1(C-12),106.9,(C-1′′′′),105.0(C-1′),101.1(C-1′′′),95.2(C-1′′),84.0(C-4′′′),83.3(C-3),78.1(C-3′′′′),77.6 ,76.6 ,76.1,75.6,75.,74.6(C-16),74.0,73.6,73.3,72.7,72.2,71.9,71.0,68.7,67.3,56.1,50.0,48.1,42.8,42.3(C-18),41.8(C-6′),41.1,39.6,37.3,37.1,36.8(-NHCOCH2-),36.5,36.4,33.6,33.4,33.1,32.0,31.3,30.8,30.7,30.6,30.5,27.3,27.2,26.0,24.8,24.5,23.8,21.6,18.3(C-6′′′),17.7,16.5(C-6′′),16.3,14.6(carbon chain terminal -CH3),10.6 ppm; HRMS. + (ESI-TOF)C 63 H 104 NO 22 [M+H] + The theoretical value was 1226.7059 and the measured value was 1226.7059. [ka]

[0194] 3-O-{[9-(4-phenoxyl-phenyl)nonanamido]-6-deoxy-β-D-glucopyranosyl}-28-O-[β- D-xylopyranosyl-(1→4)-α-L-rhamneuranosyl-(1→2)-β-D-fucopyranosyl] chelate. (122c)

[0195] 121c (20 mg, 15 μmol) and 10% Pd(OH) 2 A suspension of 5 mg / C (4 μmol) in 1.5 mL of THF / MeOH (4 / 1) was heated at 1 atm of H 2 The reaction mixture was stirred at room temperature for 12 hours. 2 Cl 2 To a stirred solution in MeOH (0.5 mL) was added pre-cooled TFA / H2O = 4 / 1 (0.5 mL) at 0 °C and stirred for 30 min. The solvent was evaporated under reduced pressure (<1 torr) at 0 °C and dried under high vacuum at room temperature for 1 h. To a stirred solution of the residue in MeOH (1 mL) was added K 2 CO 3 (40 mg, 300 μmol) was added and stirred for 12 h. The suspension was filtered, concentrated and purified by HPLC to give the product 122c (6.2 mg) as a white solid in 30% yield (HPLC column: SUPELCO Ascentis C18 25 cm×10 mm, 5 μm; mobile phase: 20% ACN / H 2 O gradient ~ 90%ACN / H 2 O for 25 min, then 90% ACN / H 2 O isocratic for 15 min; flow rate: 5 mL / min): 1H NMR(600 MHz,CD 3 OD)δ 9.41(s,1H,H-23),7.32(t,J=8.2 Hz,2H),7.17(d,J=8.4 Hz,2H),7.06(t,J=7.5 Hz,1H),6.94(d,J=8.2 Hz,2H),6.90(d,J=8.2 Hz,2H),5.40(br.s,1H,H-1′′′),5.31(br. s,1H,H-12),5.28(d,J=8.2 Hz,1H,H-1′′),4.50-4.46(m,2H,H-16,H-1′′′′),4.15(d,J=7.8 Hz,1H,H-1′),3.92-3.90(m,1H,H-2′′′),3.87-3.79(m,5H),3.67-3.64(m,2H),3.60-3.56(m,1H,H-6 a ′),3.56-3.53(m,2H),3.49-3.42(m,1H,H-4′′′′),3.36-3.33(m,1H),3.30-3.16(m,5H),3.09-3.05(m,2H),2.97 -2.92(m,1H,H-18),2.61(t,J=7.8 Hz,2H,C H 2 PhOPh),2.30(m,1H,H-19 a ),2.21(t,J=7.7 Hz,2H,-NHCOC H 2 -),1.98-1.88(m,5H),1.80-1.69(m,4H),1.67-1.59(m,4H),1.55-1.43(m,3H),1.40(s,3H,H-27),1.39-1.33(m,10H),1.31(d,J=6.2 Hz,3H,H-6′′′,),1.29(s,1H),1.20(d,J=6.4 Hz,3H,H-6′′),1.13(s,3H),1.10-1.03(m,2H),1.00(s,3H),0.98-0.94(m,1H),0.92(s,3H),0.85(s,3H),0.77(s,3H); BBD 13 C NMR(150 MHz,CD 3 OD)δ 209.4(C-23),177.2(C-28),176.7(アミド NH CO),159.3,156.5,155.1,144.9(C-13),139.2,130.8,124.0,123.1(C-12),120.0,119.4,10 7.0,(C-1′′′′),104.9(C-1′),101.1(C-1′′′),95.2(C-1′′),84.2,83.3(C-3),78.2,77.7( C-3′),76.6,76.2,75.6,75.3,74.6(C-16),74.0,73.6,73.3,72.7,72.3,71.9,71.1,68.8, 67.2,56.1,50.1,48.1,42.8,42.4(C-18),41.9(C-6′),41.2,39.6,37.1,37.3,36.5(-NHCO C H 2 -), 36.5, 36.2(- C H 2 PhOPh),33.6,33.4,32.8,31.9,31.3,30.7,30.5,30.3,27.3,27.2,26.0,25.0,24.6,21.6,,18.3(C-6′′′),17.8,16.5(C-6′′),16.4,10.6 ppm; HRMS + (ESI-TOF)C 74 H 110 NO 23 [M+H] + The theoretical value was 1380.7463 and the measured value was 1380.7476.

[0196] Experimental Example I: Immunological evaluation of saponins

[0197] Materials and Methods

[0198] Adjuvant Stock

[0199] Sample powder was dissolved in DMSO to 20 mg / mL. Prior to administration, stocks were diluted to 0.5 mg / mL with 0.5% (w / w) Tween 20 and filtered through PTFE (0.1 μm). PEK lyophilized cake (1 mg / mL PEK and 0.5 mg / mL adjuvant) or OVA (100 mg) were reconstituted, with PBS as placebo.

[0200] Animals and inoculations

[0201] C57BL / 6 mice were obtained from NLAC Taiwan. Mice aged 4-8 weeks were vaccinated with 100 μL via the subcutaneous (SC) route, once a week for 3 weeks. One week after the third vaccination, mice were sacrificed and serum and splenocytes were collected.

[0202] Splenocyte sample preparation and flow cytometry analysis

[0203] Spleen tissue was isolated from mice and processed into a single cell suspension using a sample pestle in a PP microcentrifuge. Splenocytes were cultured in 6-well plates at 2 × 10 7 / 2mL inoculation, HPV 16 Stimulated with or without E7 peptide and CO 2 The cells were then cultured in an incubator for 2 hours. After 2 hours, the cells were treated with the protein transport inhibitors monensin (Invitrogen, Catalog No. 00-4505-51) and brefeldin (Invitrogen, Catalog No. 00-4506-51) for 4 hours at 37°C. The cells were then harvested, washed twice with PBS, and stained for surface CD3 (BioLegend, Catalog No. 100290), CD4 (Invitrogen, Catalog No. 56-0041-82), and CD8 (Invitrogen, Catalog No. 12-0081-82) for 30 minutes at 4°C. After washing, cells were fixed using IC fixation buffer (Invitrogen, Cat. No. 00-8222-49) for 30 min at room temperature, then cells were washed with permeabilization buffer (Invitrogen, Cat. No. 00-8333-56) and stained with IFNγ antibody (Invitrogen, Cat. No. 53-7311-82). 20 million cell events were acquired on a Backman Coulter Gallions. Flow data were analyzed using Kaluza software (ver. 1.2). Populations were first divided into CD3 + The cells were gated on T cells and then on viable mononuclear cells using forward and side scatter. + / IFNγ+ Double positive cells or CD8 + / IFNγ + Double positive cells were subgated into either one.

[0204] ELISpot

[0205] IFNγ and IL2 production were assessed using MabTech Mouse IFNγ ELISpot PLUS Kit (3321-4HPW-2) and IL2 ELISpot PLUS Kit (3441-4HPW-2). Cells were pre-plated overnight with capture antibodies according to the manufacturer's instructions. Spleen cells were isolated from vaccinated animals and subjected to red blood cell lysis. Cells were then cultured at 2 × 10 6 Cells were resuspended at 100 μL / mL and 100 μL of cells were combined with 100 μL of stimulation master mix. The master mix contained 10 μg / mL HPV16E7-pET32a, 2 μg / ml HPV16E7-peptide. Cells were incubated in ELISpot plates at 37°C for 24 h and ELISpot assays were performed according to the manufacturer's instructions. Plates were analyzed using AID vSpot Spectrum. Values ​​were calculated by averaging triplicate wells.

[0206] ELISA

[0207] PEK was plated in 96-well plates (1 μg / well, Nunc Maxisorb) in 100 mM carbonate buffer overnight at 4°C. Plates were blocked with blocking buffer (5% milk in PBS) for at least 1 h at 37°C and then washed with PBS+0.05% Tween20. Serial two-fold dilutions of serum samples were added to the plates. After 1 h, plates were washed with PBS+0.05% Tween20 and secondary antibodies were added. Both peroxide-labeled goat anti-mouse IgG1 (Southernbiotech, Cat. No. 1070-05) and goat anti-mouse IgG2b (Southernbiotech, Cat. No. 1090-05), diluted 1:4000 in 1% milk-PBS, were added separately for 1 h. Plates were washed with PBS + 0.05% Tween 20 and developed with TMB chromogen solution (Invitrogen, Cat. No. 00-2023) for 15 min, followed by stop solution (0.2 N H 2 SO 4 ) and the absorbance at 405 nm was recorded.

[0208] Preparation of splenocyte samples and flow cytometry analysis (memory T cells)

[0209] Spleen tissue was isolated from mice and processed into a single cell suspension using a sample pestle in a PP microcentrifuge. Splenocytes were cultured in 6-well plates at 2 × 10 7 Seed with 2 mL of CO 2The cells were then cultured in an incubator for 2 hours. After 2 hours, they were stimulated with or without E7-peptide for 2 hours at 37°C, and the cells were treated with protein transport inhibitors monensin (Invitrogen, Cat. No. 00-4505-51) and brefeldin (Invitrogen, Cat. No. 00-4506-51) for 4 hours at 37°C. The cells were then harvested, washed twice with PBS, and stained for surface CD3 (BioLegend, Cat. No. 100222), CD4 (BioLegend, Cat. No. 100540), CD8 (Invitrogen, Cat. No. 11-0081-86), CD44 (BioLegend, Cat. No. 103008), and CD62L (BioLegend, Cat. No. 104412) for 30 minutes at 4°C. After washing, cells were fixed using IC Fixation Buffer (Invitrogen, Cat. No. 00-8222-49) for 30 min at room temperature, then cells were washed with Permeabilization Buffer (Invitrogen, Cat. No. 00-8333-56), IFNγ antibody (Invitrogen, Cat. No. 48-7311-82), IL-2 antibody (Invitrogen, Cat. No. 25-7021-82) and TNFα antibody (Invitrogen, Cat. No. 48-7321-82). 15 million cell events were acquired on a Backman Coulter Gallions. Flow data were analyzed using Kaluza software (ver. 1.2). Populations were first gated on viable mononuclear cells using forward and side scatter, then CD3 + / CD4 + or CD3 + / CD8 + Gated on T cells. Then sub-gated CD62L- / CD44 + Memory T cells. Next, CD4 + / IFNγ + , CD8 + / IFNγ + , CD4 + / IL-2 + , CD8 + / IL-2 + , CD4 + / TNFα + and CD8 + / TNFα+ Double positive cells were subgated into either one.

[0210] result

[0211] A mouse vaccination model was applied with the antigen PE-E7-K3 (PEK) and a fusion protein consisting of Pseudomonas exotoxin, human papillomavirus protein E7 (HPV16 E7) and KDEL3 peptide sequences, which were used to evaluate the adjuvant effect. Five mice per group were immunized three times at 3-week intervals with 50 μg of saponin and PEK (100 μg). The ability of saponin and GPI-0100 as a positive control to modulate the immune response was then analyzed by flow cytometry, ELISpot, and ELISA.

[0212] Specific T cell activation

[0213] One week after the third dose, splenocytes were harvested from the mice, and the effects of these saponin adjuvants containing the PEK antigen on the production of cytokines (IFNγ and IL-2) were measured by ELISpot (Figure 1). Saponin adjuvants 56, 63, and 79 significantly enhanced the secretion of the cytokine IFNγ, which was 3-4 times higher than GPI0100, and the induction of IL-2 was also moderate (Figure 1).

[0214] T cell activation

[0215] Antigen-specific T cell activation was analyzed by flow cytometry. One week after the third dose, splenocytes were harvested from the mice and 20 million cell events were acquired on Backman Coulter Gallions. The populations were initially CD3 + The cells were gated on T cells and then on viable mononuclear cells using forward and side scatter. + / IFNγ + (or TNFα) double positive cells or CD8 + / IFNγ + (or TNFα) double positive cells were subgated.

[0216] Based on the flow cytometry results (Figures 2-4), the aliphatic chain-modified saponins inhibited PEK-specific CD4 expression in comparison with GPI-0100. + and CD8 + The terminal aryl-substituted saponins 56, 62, and 79 potently stimulated PEK-specific IFN-γ-secreting CD8 T cells, but not GPI-0100. + and TNF-α secreted CD8 + Induced a 4-8 fold increase in T cell proliferation. However, saponin-induced CD4 + The induction of T cells was not significant. These results indicate that these saponins of the present invention are mainly CD8 + These results suggest that it mediates T cell immunity.

[0217] Memory T cell stimulation

[0218] Naive and activated T cells are known to express different adhesion molecules that are believed to display different migration patterns resulting from the expression of distinct adhesion molecules. Two adhesion molecules associated with the differentiation of naive and activated / memory T cells are CD62L (L-selectin) and CD44 (H-CAM). It has been demonstrated that naive T cells express a high CD62L and low CD44 phenotype, whereas memory T cells exhibit a low CD62L and high CD44 phenotype. Viable CD8 T cells expressing IFN-γ, TNF-α, or IL-2 are + or CD4 + Flow cytometric analysis of T cells as the frequency of splenocytes (individual mice). Cytokine positivity was determined. By flow cytometric analysis, sapoins 56, 62, and 79 significantly reduced the number of CD8 cells positive for IFN-γ or TNF-α compared to mice vaccinated with GPI-0100. + A high frequency of T cells was confirmed (Figures 5 and 6). Of note, cytokine-positive CD4 + No T cells were detected. These results indicated that 56, 62, and 79 could provide long-term cellular immune protection against the E7 antigen.

[0219] Antibody production assay

[0220] Serum PEK (coated with E7)-specific IgG antibody titers were determined using ELISA after each administration. As shown in Figure 7, PEK / GPI-0100 induced the highest level of antibody production in C57BL / 6 mice. Among them, compounds 53-56 could induce moderate E7-specific antibody production. Because cellular immunity and humoral immunity are mutually inhibitory, it was reasonable that the compounds induced higher cytotoxic T cell immunity with lower antibody production.

[0221] Immunological analysis of our compounds after vaccination suggested that compounds 46–62, 64, 66, 77–79, 83, 92, and 95 are potent saponin-based adjuvants for developing cellular immunity against the host. Adjuvants with these properties would be advantageous in combination with therapeutic vaccines against cancer, bacteria (tuberculosis), viruses (HIV, herpes), protozoa (malaria), etc.

[0222] toxicity

[0223] Acute toxicity was investigated with increasing doses of Saponin 56 from 100 μg to 1000 μg. Results were presented as the number of surviving animals per group of 5 mice (female BALB / c mice, 9 weeks) over a 7-day period. After this test, all mice survived and had no obvious abnormalities in their activity and feeding behavior. (Figure 8) [Table 1]

[0224] result

[0225] The results are shown in Figure 8, which shows that the median percentage body weight change in mice receiving increasing doses of saponin adjuvant 56 was all less than 5%. Spleen somatic cell index and liver somatic cell index in all experimental groups were unchanged compared to the control group. These data suggest that saponin 56 is a potent and safe candidate as a vaccine adjuvant.

[0226] Experimental Example II: Tumor challenge with OVA peptide vaccine

[0227] Materials and Methods

[0228] Adjuvant Stock

[0229] The sample powder was dissolved in DMSO to a concentration of 20 mg / mL.

[0230] Animals and inoculations

[0231] Six- to eight-week-old female C57BL / 6 mice were obtained from NLAC Taiwan. Mice were inoculated with 200 μL of 1.5 × 10 6 The tumor volumes were measured periodically using a caliper and calculated by the following formula:

number

[0232] result

[0233] To evaluate the antitumor effect, the antigen OVA was applied in a mouse vaccination model. Five mice per group were immunized twice with 50 μg saponin and 100 μg OVA at 3-week intervals. The antitumor effect of compound 56 and the positive control groups (alum, Qs-21 and GPI-0100) were analyzed by caliper.

[0234] Antitumor effects

[0235] E.G7-OVA tumor-bearing mice were vaccinated intradermally twice with different formulations on days 7 and 14. Compared with the control group (PBS), mice treated with OVA and OVA+Alum initially showed slight tumor growth inhibition, but the treatment was not effective enough and rapid tumor growth resumed later. In contrast, mice treated with OVA+Compound 56, OVA+Qs-21, and OVA+GPI0100 showed significant tumor growth inhibition effects. Moreover, OVA+Compound 56 had the highest survival rate (Figure 9).

[0236] Experimental Example III: Influenza challenge with OVA peptide vaccine.

[0237] Materials and Methods

[0238] Adjuvant Stock

[0239] The sample powder was dissolved in DMSO to a concentration of 20 mg / mL.

[0240] Animals and inoculations

[0241] Female C57BL / 6 mice aged 6-8 weeks were either immunized by subcutaneous injection with 100 μl of vaccine or by intranasal administration with 30 μl of vaccine. All vaccine solutions were freshly prepared and diluted in 0.5% Tween 20 + PBS. The vaccine used for subcutaneous injection contained the immunogen NP 366-374 / NP 311-325The peptide was included alone or in combination with Compound 56 (50 μg). Vaccines were administered intranasally and included the immunogen alone or with Compound 56 (30 μg). After two vaccinations, mice were intranasally infected with 110 plaque-forming units (PFU) of live PR8 virus.

[0242] result

[0243] A mouse vaccination model was applied with the antigen OVA to evaluate the anti-influenza efficacy. Five mice per group were vaccinated with compound 56 and NP. I / II After the second vaccination, mice were intranasally infected with 110 plaque-forming units (PFU) of live PR8 virus. I and NP II (NP I / II ) compared to mice immunized with peptide alone, NP I / II and compound 56 immunized mice had increased survival rates after PR8 infection (Figure 10).

[0244] Experimental Example IV: Immunological evaluation of the combination of SARS-CoV-2 antigens and Compound 56 immune adjuvant

[0245] Materials and Methods

[0246] Adjuvant Stock

[0247] Sample powder was dissolved in DMSO to a concentration of 20 mg / mL. Before administration, the stock was diluted to 0.5 mg / mL with 0.5% (w / w) Tween 20 and filtered through PTFE (0.1 μm).

[0248] Animals and inoculations

[0249] Female C57BL / 6 mice aged 6–8 weeks were vaccinated by the subcutaneous (SC) route with SARS-CoV-2 spike protein and alum or compound 56, administered three times at 6-week intervals. Blood was collected from the tail artery of the mice 10 days after each immunization to measure IgG levels.

[0250] ELISA

[0251] The levels of specific serum IgG against the SARS-CoV-2 spike protein in each group were measured using borate buffered saline (BBS; 100 mM NaCl, 50 mM boric acid, 1.2 mM Na 2 B 4 O 7 The serum levels were measured by ELISA using Maxisorp microtiter plates (NUNC International, Roskilde, Denmark) coated with SARS-CoV-2 spike RBD His protein (0.5 μg / well) in 1% BSA-PBS (pH 8.2) overnight at 4 °C. Plates were blocked with blocking buffer (5% milk in PBS) for at least 1 h at 37 °C and then washed with PBS + 0.05% Tween 20. Serial 5-fold dilutions of serum samples were added to the plate. After 1 h, plates were washed with PBS + 0.05% Tween 20. Peroxide-labeled goat anti-mouse IgG (Invitrogen, Cat. No. 81-6520) diluted 1:3000 in 1% BSA-PBS was added for 1 h. Plates were washed with PBS + 0.05% Tween 20 and developed with TMB chromogen solution (Invitrogen, Cat. No. 00-2023) for 15 min followed by stop solution (0.2 N H 2 SO 4 ) and the absorbance at 450 nm was recorded.

[0252] result

[0253] Antigen SARS-CoV-2 spike RBD His protein was administered to a mouse vaccination model to evaluate its anti-COVID19 efficacy. Mice were divided into three groups: Compound 56 (50 μg), SARS-CoV-2 (2 μg) + Compound 56 (50 μg), and SARS-CoV-2 (10 μg) + Alum (10 μg).

[0254] Antibody production assay

[0255] Serum SARS-CoV-2-specific IgG antibody titers were determined using ELISA after each dose. As shown in Figure 11, the SARS-CoV-2 + Compound 56 group induced the highest level of antibody production in C57BL / 6 mice. Among them, the SARS-CoV-2 / Compound 56 group showed a 1000-fold increase in antibody titers with only 20% of the antigen. Yet another aspect of the present invention may be as follows. [1] A saponin conjugate represented by formula (I) or a pharma- ceutically acceptable salt thereof. TIFF0007680966000101.tif3389 (I) (where: TIFF0007680966000102.tif39 is a single bond or a double bond, W is a methyl group (Me), -CHO, TIFF0007680966000103.tif1932 , -CH 2 OH or -CH 3 and V is H or OH; Y is CH 2 , -O-, -S-, -NR-, or -NH-; Q is CH 2 , C=O, C=N-OH, or C=N-OMe; X is CH 2 , -O-, -NR-, -NH-(C=O)-, -S-, or O-(C=O)-; R is a cyclic or acyclic, optionally substituted moiety selected from the group consisting of acyl, aliphatic, heteroaliphatic, aryl, arylaliphatic, cycloaliphatic, heterocyclic aliphatic, heteroarylaliphatic, alkyloxyaliphatic, and aryloxyaliphatic, or C 1~C 18 an optionally substituted moiety selected from the group consisting of aliphatic, 5-10 membered arylaliphatic, 5-10 membered heteroarylaliphatic having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and 4-7 membered heterocyclylaliphatic having 1-2 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur; R 1 are independently hydrogen, an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates and carbonates, or a carbohydrate having a monosaccharide structure; Z is a linear or branched oligosaccharide or an optionally substituted group selected from the group consisting of amine, amide, acyl, arylalkyl, aryl, heteroaryl, aliphatic, heteroaliphatic, cycloaliphatic, and heterocyclyl. [2] The saponin conjugate or its pharma- ceutically acceptable salt according to [1], wherein R is a heteroaliphatic, an arylaliphatic, a heterocyclic aliphatic, a heteroarylaliphatic, an alkyloxyaliphatic, or an aryloxyaliphatic. [3] The saponin conjugate or pharma- ceutically acceptable salt thereof according to [1] above, wherein Z is a linear tetrasaccharide or a linear trisaccharide, and the first sugar residue is directly linked to Y. [4] The saponin conjugate or a pharma- ceutically acceptable salt thereof according to [1] above, wherein W is CHO and V is OH. [5] The saponin conjugate or a pharma- ceutically acceptable salt thereof according to [1] above, wherein Q is C=O and X is -NH-. [6] Q is CH 2 and X is OC(=O), or a pharma- ceutically acceptable salt thereof, according to [1] above. [7] The saponin conjugate or a pharma- ceutically acceptable salt thereof according to [2] above, wherein R is alkyl, e.g., dodecyl, methyl, hexyl, octadecyl, ethyl, propyl, pentyl, etc. TIFF0007680966000104.tif85170 [8] The saponin conjugate according to [1] above, having the following structure: TIFF0007680966000105.tif80165 [9] A vaccine composition comprising an antigen and the saponin conjugate according to [1] or a pharma- ceutically acceptable salt thereof.

[10] The vaccine composition according to [9] above, further comprising an additional adjuvant.

[11] The vaccine composition of [9] above, further comprising a pharma- ceutically acceptable carrier or diluent.

[12] The vaccine composition according to [9], wherein the antigen is selected from the group consisting of a bacterial antigen, a virus-associated antigen, and a tumor-associated antigen.

[13] The bacterial antigen is selected from the group consisting of Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Borrelia burgdorferi, Borrelia spp., Chlamydia trachomatis, Helicobacter pyloris, Chlamydia pneumoniaes, Ureaplasma urealyticums, Mycoplasma pneumoniaes, Staphylococcus spp., Staphylococcus aureus, Streptococcus pyogenes, and the like. pyogenes, Streptococcus spp., Streptococcus pneumoniaes, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis, Neisseria gonorrhoeae, Bacillus anthracis, Salmonella spp., Salmonella typhi, Vibrio cholera, Pasteurella pestis, Campylobacter spp., Campylobacter jejuni, Clostridium spp. spp.), Clostridium difficile, Corynebacterium diphtheria, Mycobacterium spp.), Mycobacterium tuberculosis, Pseudomonas aeruginosa, Treponema spp., Leptospria spp., Hemophilus ducreyi, Hemophilus influenza, Escherichia coli, Shigella spp., Erlichia spp., Rickettsia spp., and combinations thereof. The vaccine composition according to

[12] above, wherein the antigen is associated with a bacterium selected from the group consisting of Escherichia coli, Shigella spp., Erlichia spp., Rickettsia spp., and combinations thereof.

[14] The vaccine composition according to

[12] , wherein the virus-related antigen is an antigen related to a virus selected from the group consisting of influenza virus, parainfluenza virus, mumps virus, adenovirus, respiratory syncytial virus, Epstein-Barr virus, rhinovirus, poliovirus, coxsackievirus, echovirus, measles virus, rubella virus, varicella-zoster virus, herpes virus, herpes simplex virus, parvovirus, cytomegalovirus, hepatitis virus, human papillomavirus, alpha virus, flavivirus, bunyavirus, rabies virus, arenavirus, filovirus, HIV1, HIV2, HTLV-1, HTLV-II, FeLV, bovine LV, FeIV, canine distemper virus, canine infectious hepatitis virus, feline calicivirus, feline rhinotracheitis virus, TGE virus, foot and mouth disease virus, coronavirus, dengue virus, fabivirus, and combinations thereof.

[15] The tumor-associated antigen is selected from the group consisting of killed tumor cells and their lysates, MAGE-1, MAGE-3 and their peptide fragments, human chorionic gonadotropin and its peptide fragments, carcinoembryonic antigen and its peptide fragments, α-fetoprotein and its peptide fragments, pancreatic carcinoembryonic antigen and its peptide fragments, prostate-specific antigen and its peptide fragments, MUC-1 and its peptide fragments, CA125, CA15-3, CA19-9, CA549, CA195 and their peptide fragments, prostate-specific membrane antigen and its peptide fragments, squamous cell carcinoma antigen and its peptide fragments, ovarian cancer antigen and its peptide fragments, pancreatic cancer-associated antigen and its peptide fragments, Her1 / neu and its peptide fragments, gp-100 and its peptide fragments, mutant K-ras protein and its peptide fragments, mutant p53 and its peptide fragments, truncated epidermal growth factor receptor, and chimeric protein p210. BCR-ABL ,STn,Tn,Lewis x , Lewis y The vaccine composition according to claim 12, wherein the antigen is selected from the group consisting of TF, GM1, GM2, GD2, GD3, Gb3, KH-1, Globo-H, SSEA-4, and mixtures thereof.

[16] A pharmaceutical composition comprising one or more of the saponin conjugates according to [1] above or pharma- ceutically acceptable salts thereof.

[17] The pharmaceutical composition according to

[16] above, further comprising a pharma- ceutically acceptable excipient.

[18] A saponin conjugate intermediate represented by formula (II) or a pharma- ceutically acceptable salt thereof. TIFF0007680966000106.tif3179 (II), [where: TIFF0007680966000107.tif39 is a single bond or a double bond, W is a methyl group (Me), -CHO, TIFF0007680966000108.tif1732 , -CH 2 OR 1 , -C(O)R, or CH 2 OR x and V is hydrogen or -OR 1 and Y is CH 2 , -O-, -NR-, -NH-, or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; Q is CH 2 , C=O, C=N-OH, or C=N-OMe; X is CH 2 , -O-, -NR-, -NH-(C=O)-, -S-, O-(C=O)-, CH 2 , -O-, -NR-, or -NH-(C=O)-, -S-, or O-(C=O)-; R 1 is independently hydrogen, an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates and carbonates, or a carbohydrate having the structure: TIFF0007680966000109.tif3173 (where: a, b, and c, independently in each occurrence, are 0 or 1; R 0 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R a 、R b 、R c , and R d is independently in each occurrence hydrogen, halogen, OH, OR, OR x and R x is independently in each occurrence hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R is a cyclic or acyclic, optionally substituted moiety selected from the group consisting of acyl, aliphatic, heteroaliphatic, aryl, arylaliphatic, cycloaliphatic, heterocyclic aliphatic, heteroarylaliphatic, alkyloxyaliphatic, and aryloxyaliphatic, or C 1 ~C 18 an optionally substituted moiety selected from the group consisting of aliphatic, 5-10 membered arylaliphatic, 5-10 membered heteroarylaliphatic having 1-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and 4-7 membered heterocyclylaliphatic having 1-2 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur.

[19] A saponin conjugate intermediate of formula (II) according to

[18] above or a pharma- ceutically acceptable salt thereof, obtained by reacting a compound represented by the structure of formula (III) with a compound represented by the structure of formula (IV) or a pharma- ceutically acceptable salt thereof. TIFF0007680966000110.tif2749 (III) [where: R 1 is independently hydrogen, an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates and carbonates, or a carbohydrate having the structure: TIFF0007680966000111.tif2966 (where: a, b, and c, independently in each occurrence, are 0 or 1; R 0 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R a 、R b 、R c , and R d is independently in each occurrence hydrogen, halogen, OH, OR, OR x and R x is independently in each occurrence hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; LG is a leaving group, TIFF0007680966000112.tif1629 or TIFF0007680966000113.tif1728 )] TIFF0007680966000114.tif3561 (IV) [where: TIFF0007680966000115.tif38 is a single bond or a double bond, W is a methyl group (Me), -CHO, TIFF0007680966000116.tif1834 , -CH 2 OR or -C(O)R; V is hydrogen or -OR 1 and Y is CH 2 , -O-, -S-, -NR-, -NH-, -S-, or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R, R x or R 1 are independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates.

Claims

1. A saponin conjugate of formula (I) or a pharma- ceutically acceptable salt thereof. 【Chemistry 1】 (where: 【change】 is a single bond or a double bond, W is a methyl group (Me), -CHO, 【change】 , -CH 2 OR x or -C(=O)R y where R x are independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R y is selected from the group consisting of alkyl, benzyl, silyl, alkoxy and alkyl carbonate; V is —H or —OH; Y is -CH 2 -, -O-, -S-, -NR-, or -NH-; Q is -CH 2 -, -C(=O)-, -C(=N-OH)-, or -C(=N-OMe)-, X is -CH 2 -, -O-, -NH-, -NH-C(=O)-, -S-, or -O-C(=O)-, R is 【Chemistry 2】 【change】 【change】 【change】 and 【change】 Selected from the group consisting of: where Rz is alkyl. In addition, Q is -CH 2 -, X is -O-(C=O)-, and R belongs to an arylaliphatic group or aryl, then Q-X-R is a configuration represented by the formula: Q-(C=O)-O-R, R 1 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; Z is a linear or branched oligosaccharide optionally substituted with a group selected from the group consisting of amine, amide, acyl, arylalkyl, aryl, heteroaryl, aliphatic, heteroaliphatic, cycloaliphatic, and heterocyclyl.

2. 2. The saponin conjugate of claim 1, or a pharma- ceutically acceptable salt thereof, wherein Z is a linear tetrasaccharide or a linear trisaccharide, and the first sugar residue is directly linked to Y.

3. 2. The saponin conjugate of claim 1, or a pharma- ceutically acceptable salt thereof, wherein W is -CHO and V is -OH.

4. 2. The saponin conjugate of claim 1, or a pharma- ceutically acceptable salt thereof, wherein Q is -C(=O)- and X is -NH-.

5. Q is -CH 2 - and X is -O-C(=O)-, or a pharma- ceutically acceptable salt thereof.

6. 2. The saponin conjugate of claim 1 having the structure: 【Chemistry 3】

7. 10. A vaccine composition comprising an antigen and the saponin conjugate of claim 1 or a pharma- ceutically acceptable salt thereof.

8. The vaccine composition of claim 7, further comprising an additional adjuvant.

9. 8. The vaccine composition of claim 7, further comprising a pharma- ceutically acceptable carrier or diluent.

10. 8. The vaccine composition of claim 7, wherein the antigen is selected from the group consisting of a bacterial antigen, a virus-associated antigen and a tumor-associated antigen.

11. The bacterial antigens include Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Borrelia burgdorferi, Borrelia spp., Chlamydia trachomatis, Helicobacter pylori, Chlamydia pneumoniae, Ureaplasma urealyticum, and the like. urealyticums, Mycoplasma pneumoniae, Staphylococcus spp., Staphylococcus aureus, Streptococcus pyogenes, Streptococcus spp., Streptococcus pneumoniae, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis meningitidis, Neisseria gonorrhoeae, Bacillus anthracis, Salmonella spp., Salmonella typhi, Vibrio cholera, Pasteurella pestis, Campylobacter spp., Campylobacter jejuni, Clostridium spp., Clostridium difficile, difficile, Corynebacterium diphtheria, Mycobacterium spp., Mycobacterium tuberculosis, Pseudomonas aeruginosaaeruginosa, Treponema spp., Leptospura spp., Hemophilus ducreyi, Hemophilus influenza, Escherichia coli, Shigella spp., Erlichia spp., Rickettsia spp., and any combination thereof.

12. The virus-related antigens include influenza virus, parainfluenza virus, mumps virus, adenovirus, respiratory syncytial virus, Epstein-Barr virus, rhinovirus, poliovirus, coxsackievirus, echovirus, measles virus, rubella virus, varicella-zoster virus, and the like.

11. The vaccine composition of claim 10, wherein the antigen is an antigen associated with a virus selected from the group consisting of a virus, a herpes virus, a herpes simplex virus, a parvovirus, a cytomegalovirus, a hepatitis virus, a human papilloma virus, an alpha virus, a flavivirus, a bunyavirus, a rabies virus, an arenavirus, a filovirus, HIV1, HIV2, HTLV-1, HTLV-II, FeLV, bovine LV, FeIV, canine distemper virus, infectious canine hepatitis virus, feline calicivirus, feline rhinotracheitis virus, TGE virus, foot and mouth disease virus, a coronavirus, a dengue virus, a fabivirus, and any combination thereof.

13. The tumor-associated antigens include killed tumor cells and their lysates, MAGE-1, MAGE-3 and their peptide fragments, human chorionic gonadotropin and its peptide fragments, carcinoembryonic antigen and its peptide fragments, alpha-fetoprotein and its peptide fragments, pancreatic carcinoembryonic antigen and its peptide fragments, prostate-specific antigen and its peptide fragments, MUC-1 and its peptide fragments, CA125, CA15-3, CA19-9, CA549, CA195 and their peptide fragments, prostate-specific membrane antigen and its peptide fragments, squamous cell carcinoma antigen and its peptide fragments, ovarian cancer antigen and its peptide fragments, pancreatic cancer associated antigen and its peptide fragments, Her1 / neu and its peptide fragments, gp-100 and its peptide fragments, mutant K-ras protein and its peptide fragments, mutant p53 and its peptide fragments, truncated epidermal growth factor receptor, chimeric protein p210 BCR-ABL , STn, Tn, Lewis x , Lewis y 11. The vaccine composition of claim 10, wherein the antigen is selected from the group consisting of TF, GM1, GM2, GD2, GD3, Gb3, KH-1, Globo-H, SSEA-4, and any mixture thereof.

14. 10. A pharmaceutical composition comprising one or more saponin conjugates of claim 1 or pharma- ceutically acceptable salts thereof.

15. 15. The pharmaceutical composition of claim 14, further comprising a pharma- ceutically acceptable excipient.

16. A saponin conjugate intermediate represented by formula (II) or a pharma- ceutically acceptable salt thereof. 【Chemistry 4】 [where: 【change】 is a single bond or a double bond, W is a methyl group (Me), -CHO, 【change】 , -C(=O)R y , or -CH 2 OR x where R x are independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R y is selected from the group consisting of alkyl, benzyl, silyl, alkoxy and alkyl carbonate; V is -H or -OR x and Y is -CH 3 , -OH, -SH, -OR 5 or -NH 2 where OR 5 is selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; Q is -CH 2 -, -C(=O)-, -C(=N-OH)-, or -C(=N-OMe)-, X is -CH 2 -, -O-, -NH-, -NH-C(=O)-, -S-, or -O-C(=O)-, R 1 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates. R is 【Chemistry 5】 【change】 【change】 【change】 and 【change】 and Rz is alkyl; Here, Q is -CH 2 -, X is -O-(C=O)-, and R belongs to an arylaliphatic group or aryl, then Q-X-R is a configuration represented by the formula: Q-(C=O)-O-R.

17. 17. A method for preparing the saponin conjugate intermediate of formula (II) or a pharma- ceutically acceptable salt thereof according to claim 16, comprising reacting a compound represented by the structure of formula (III) with a compound represented by the structure of formula (IV) or a pharma- ceutically acceptable salt thereof to obtain the saponin conjugate intermediate of formula (II) or a pharma- ceutically acceptable salt thereof. 【Chemistry 6】 [where: R 1 is independently hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates. L.G. leaving group, 【change】 or 【change】 )] 【Chemistry 7】 [where: 【change】 is a single bond or a double bond, W is a methyl group (Me), -CHO, 【change】 , -CH 2 OR x or -C(=O)R y and V is -H or -OR x and Y is -CH 3 , -OH, -SH, -OR 5 or -NH 2 and Here, R x is hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; R y is selected from the group consisting of alkyl, benzyl, silyl, alkoxy and alkyl carbonate; R 2 is hydrogen or an oxygen protecting group selected from the group consisting of alkyl ethers having at least 5 carbon atoms, benzyl ethers, silyl ethers, acetals, ketals, esters, carbamates, and carbonates; OR 5 is a group selected from the group consisting of alkyl ether, benzyl ether, silyl ether, acetal, ketal, ester, carbamate, and carbonate.

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