CN114805454A - Alpha-galactose ceramide compound and preparation method and application thereof - Google Patents

Abstract

The invention relates to an alpha-galactosyl ceramide compound shown in a general formula (I) or a pharmaceutically acceptable salt or solvate thereof, and also relates to a preparation method of the compound and application of the compound in preparing a medicament capable of inducing Th1 selective immune response.

Description

Alpha-galactose ceramide compound and preparation method and application thereof

Technical Field

The invention relates to an alpha-galactosylceramide (for example, alpha-GalCer, also known as KRN7000) compound capable of inducing selective immune response of Th1, and a preparation method and application thereof, belonging to the technical field of chemistry and medicine.

Background

Immune responses are a complex process involving a variety of cells such as B cells, T cells, Natural Killer (NK) cells, Dendritic Cells (DCs), NKT cells, macrophages, and the like in organs such as bone marrow, spleen, thymus, and the like. The lack of immune function can cause the invasion of foreign microorganisms into the matrix to cause inflammatory reaction or the canceration of self cells to cause cancer, and the over-strong immune function can cause multiple sclerosis, rheumatism, type II diabetes and other autoimmune diseases. Therefore, it is important to intervene in immune response timely to keep its normal function.

Natural killer T cells (NKT cells) are a special subset of T cells that have both Natural Killer (NK) cell and T cell receptors on their surface. Studies have shown that iNKT cells, a species of NKT cells, play a versatile role in the regulation of the immune system. First, such cells are stimulated to produce large amounts of a range of cytokines such as IFN- γ, IL-4, IL-5, IL-6, IL-17, etc. Second, iNKT cells produce cytokines that modulate other immune cells, such as macrophages, B cells, and some conventional T cells, among others. Furthermore, iNKT cells were able to express granzyme B, FasL, perforin, etc. at high levels and thus demonstrated cytotoxic ability.

Morita et al found that alpha-galactosylceramide compounds extracted from Agelas mauritianus sponge have anticancer activity. Tsuji et al found that α -galactosylceramide compounds, upon entering the body, were recognized by Dendritic Cells (DCs), bound to CD1d thereon, and presented to iNKT cells, bound to TCRs thereon, to produce Th 1-type cytokines and Th 2-type cytokines by activating iNKT cells. Wherein Th1 cytokines (such as IFN-gamma, TNF-alpha, IL-2 and IL-12) have pro-inflammatory effect, and correspondingly have the effects of resisting pathogenic microorganisms such as bacteria, tuberculosis, virus and the like, resisting cancers and tumors and the like; th2 cytokines (such as IL-4, IL-10, IL-5 and IL-6) have immunoregulation effect, and have relieving and treating effects on autoimmune diseases.

However, in the iNKT cells activated by α -GalCer, the Th1 cytokine and the Th2 cytokine are simultaneously produced in a large amount, and there is a mutual antagonism between the Th1 cytokine and the Th2 cytokine, for example, the Th2 cytokine can weaken the tumor cell inhibitory ability of α -GalCer, so that α -GalCer cannot be successfully applied to clinic. Research shows that by selectively inducing iNKT cells to secrete Th1 or Th2 cytokines, antagonism caused by simultaneous secretion of a large amount of the two cytokines can be avoided, and excellent effects of Th1 or Th2 cytokines on immunity can be ensured. However, how to achieve high secretion selectivity of iNKT cells for both Th1 and Th2 cytokines, thereby generating a desired immune response based on the high selectivity of Th1 or Th2 cytokines, is a technical problem to be solved in the art.

Chinese patent application No.201510765501.3 discusses the introduction of a single hydroxyl group on the amide chain of α -GalCer or two consecutive hydroxyl groups on adjacent carbons to obtain some analogs of α -GalCer that are potentially biologically active. However, the patent application does not relate to the exact position of hydroxyl groups introduced to obtain truly biologically active, even significantly superior (but not potentially biologically active) analogs of α -GalCer. Moreover, it is not easy to design and synthesize a new compound which does not exist in the prior art, and the patent application does not give corresponding confirmation data of preparation and activity, and it is not known whether the hydroxyl group can be successfully introduced into α -GalCer, and it is more difficult to know the biological activity. Therefore, there is no prior art how to structurally modify α -GalCer with hydroxyl group to obtain α -GalCer analogs with remarkably superior biological activity.

In addition, the thia sugar is a sugar compound formed by substituting an oxygen atom in a sugar ring with a sulfur atom. The replacement of the oxygen atom on the sugar ring of the alpha-GalCer with the sulfur atom is a research direction for obtaining the alpha-GalCer analogue with potential biological acquisition, however, the synthesis of the thiogalactose is difficult, the preparation process is complex and tedious, and the yield is extremely low, thereby greatly limiting the application and research of the sugar derivative.

Based on the above, the problem to be solved by the present invention is that how to specifically modify α -GalCer with hydroxyl group (for example, at which specific position of amide chain of α -GalCer the hydroxyl group is introduced) can obtain α -GalCer analog with excellent activity of inducing selective immune response of Th1, thereby enabling iNKT cells to achieve the purpose of excellent selective secretion of Th1 and Th2 cytokines. The problem to be solved by the present invention is also to provide an α -GalCer analog, such as a thiogalactoside-based 5-Thio- α -GalCer analog, respectively, which has significantly superior activity of inducing a selective immune response to Th1 compared to existing α -GalCer analogs. The present invention is an improvement over patent application No. 201510765501.3.

Disclosure of Invention

In order to solve the above technical problems, the inventors have conducted a simulation analysis of the interaction between CD1d protein and ligand by computer-aided method, and unexpectedly found that three specific polar residues of GLN-14, SER-28 and TYR-73 present in the binding domain of CD1d protein have a key effect on the binding between protein and ligand. With respect to the three specific polar residues, the inventors introduced a polar group such as a monohydroxy group or a sulfuric acid group (i.e., R) at any one of the specific positions C11, C12, or C13 of the amide chain of α -GalCer (and 5-Thio- α -GalCer) ₁ 、R ₂ And R ₃ Either of which is a hydroxyl or a sulfate group, the remaining two being hydrogen), and the introduced polar group can interact with the above-mentioned polar residues of the CD1d protein, thereby significantly enhancing the binding effect of the obtained α -GalCer analog and CD1 d. Thereby, a class capable of being remarkably increased is obtainedTh1 selectivity for strong induction of immune response, alpha-GalCer analogue with improved anticancer activity.

Specifically, the invention is realized by the following aspects:

in a first aspect, the present invention provides an α -galactosylceramide compound represented by the general formula (I) or a pharmaceutically acceptable salt or solvate thereof:

wherein, X ₁ And X ₂ Each is independently selected from O or S;

R ₁ 、R ₂ and R ₃ Either one of them is hydroxyl or sulfate, and the other two are H; and is

R ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ And R ₉ Each independently selected from hydroxyl or sulfate.

In a second aspect, the present invention provides a pharmaceutical composition comprising: a compound represented by the general formula (I) according to the first aspect or a pharmaceutically acceptable salt or solvate thereof; and pharmaceutically acceptable auxiliary materials.

In a third aspect, the present invention provides a use of the compound represented by the general formula (I) in the first aspect or a pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition in the second aspect, in the preparation of a medicament for inducing a Th1 selective immune response, wherein the medicament for inducing a Th1 selective immune response comprises an anti-tumor medicament, an anti-viral medicament, an antibacterial medicament, an anti-tuberculosis medicament, and a medicament for regulating autoimmune activity.

In a fourth aspect, the present invention provides a method for preparing a compound represented by the general formula (I) in the first aspect, or a pharmaceutically acceptable salt or solvate thereof.

Drawings

FIG. 1 shows alpha-GalC with introduction of polar groups at different positions of the amide chain of alpha-GalCer obtained by computer-assisted drug designThe fraction of binding of the er analog to the CD1d binding domain. As shown in fig. 1, the inventors found that the docking score of GalCer analogs having a hydroxyl group or a sulfate group at C11, C12, or C13 was significantly higher (more than 21 points) compared to other positions of the amide chain, suggesting that such GalCer analogs have significantly stronger binding effect with CD1 d. Wherein "GC _ 8R" represents an alpha-GalCer-OH analogue with a hydroxyl group at the C8 position of the amide chain and an R type configuration, and "GC _ 8S" represents an alpha-GalCer-OH analogue with a hydroxyl group at the C8 position of the amide chain and an S type configuration, and so on; "GCs _ 11R" represents a 5-Thio- α -GalCer-OH analog having a hydroxyl group at the C11 position of the amide chain and the configuration R; "GCs _ 11S" represents a 5-Thio- α -GalCer-OH analog having a hydroxyl group at the C11 position of the amide chain and an S-type configuration, and so on; "GC-12 rso 3" represents a-GalCer-SO having the C12 position of the amide chain being a sulfate group and the configuration being R type ₃ ^- (ii) an analog; "GC-12 sso 3" represents a-GalCer-SO in which the C12 position of the amide chain is a sulfate group and the configuration is S type ₃ ^- (ii) an analog; "GCs-12 rso 3" represents 5-Thio-alpha-GalCer-SO in which C12 position of the amide chain is a sulfate group and the configuration is R type ₃ ^- (ii) an analog; "GCs-12 sso 3" represents 5-Thio-alpha-GalCer-SO wherein the C12 position of the amide chain is sulfuric acid group and the configuration is S type ₃ ^- And the like.

FIG. 2 shows a schematic representation of the docking and interaction of α -GalCer analogs with proteins, in which polar residues such as GLN-14, SER-28 and TYR-73, present in the binding domain of CD1d protein, interact with hydroxyl or sulfuric acid introduced at specific positions (C11, C12 or C13) on the α -GalCer amide chain via hydrogen bonding, enhancing the binding between the protein and the ligand.

FIG. 3 shows the ratio of the IFN- γ and IL-4 secretion induced by PBS, α -GalCer, and exemplary compounds of the invention, the maximum secretion (i.e., peak) of IFN- γ/IL-4, and the area under the line (AUC) of the FN- γ/IL-4 secretion curve.

FIG. 4 is the results of anti-murine vaccinia using PBS, α -GalCer, and exemplary compounds of the invention. Wherein PFU represents a plaque forming unit (plaque forming unit) for use in a method for quantifying an animal virus; DPI indicates the number of infection days (days post-infection).

FIG. 5 is the results of anti-melanoma treatment with PBS, α -GalCer, and exemplary compounds of the present invention.

FIG. 6 is the results of anti-breast cancer with PBS, α -GalCer, and exemplary compounds of the present invention.

Detailed Description

It will be understood by those skilled in the art that the following embodiments are given by way of example only and are not intended to limit the scope of the present application in any way.

As used herein, unless otherwise indicated, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. For example, the term "pharmaceutically acceptable salt" as used herein refers to an acid addition salt or a base addition salt of a compound of formula (I) with a pharmaceutically acceptable free acid or free base. The acid addition salts are obtained from the following acids: such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, tartaric acid, fumaric acid, malic acid, oxalic acid, succinic acid, and the like. The base addition salt includes sodium salt, potassium salt, calcium salt, ammonium salt, magnesium salt, etc.

The solid and dashed bonds of the wedge shape are used herein, unless otherwise indicated (

And

) Indicating the absolute configuration of a stereocenter. Unless otherwise indicated, the stereoisomers mentioned herein include geometric isomers and enantiomers, all of which are within the scope of the present application.

In this context, unless otherwise indicated, use is made ofThe term "sulfate group" means-OSO ₃ H and its free form-OSO ₃ ^- 。

Herein, unless otherwise indicated, the use of the term "GalCer" or "GC" refers to the half lactose ceramide, such as alpha GalCer refers to alpha galactosyl ceramide, also known as KRN 7000. The term "GalCer analogs" is used to refer to a class of compounds that structurally modify GalCer. The term "GalCer-OH analog" or "GalCer-SO" is used ₃ ^- The analog refers to a compound of GalCer modified by hydroxyl or sulfate.

As used herein, unless otherwise indicated, the term "5-Thio-GalCer" or "GCs" refers to a thiogalactosyl ceramide analogue in which the oxygen atom of the α -GalCer sugar ring is replaced with a sulfur atom. The term "5-Thio-GalCer-OH analog" or "5-Thio-GalCer-SO" is used ₃ ^- The analog "refers to a compound of which 5-Thio-GalCer is modified by hydroxyl or sulfate.

In one embodiment, the present invention provides α -galactosylceramide compounds represented by the general formula (I):

wherein, X ₁ And X ₂ Each is independently selected from O or S;

R ₁ 、R ₂ and R ₃ Either one of them is hydroxyl or sulfate, the other two are both H; and is

R ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ And R ₉ Each independently selected from hydroxyl or sulfate.

In some embodiments, the α -galactosylceramide compound of formula (I) has the structure of formula (II) as follows:

wherein, X ₁ And X ₂ And R ₁ -R ₉ As defined above.

In some embodiments, the α -galactosylceramide compound of formula (I) has the structure shown in formula (III) below:

wherein, X ₁ And X ₂ Each is independently selected from O or S; and is

R ₁ 、R ₂ And R ₃ Either one of them is a hydroxyl group or a sulfate group, and the remaining two are both H.

In some embodiments, R ₁ 、R ₂ And R ₃ One of the following three situations A, B and C is satisfied:

(A)R ₁ is hydroxy or sulfuric acid radical, and R ₂ And R ₃ Is H;

(B)R ₂ is hydroxy or sulfuric acid radical, and R ₁ And R ₃ Is H; and

(C)R ₃ is hydroxy or sulfuric acid radical, and R ₁ And R ₂ Is H;

wherein R is a hydroxyl group ₁ 、R ₂ And R ₃ The chirality of the carbon on which either is located is R or S.

In some embodiments, the α -galactosylceramide compound of formula (I) has the structure of formula (IV) or formula (V) as follows:

wherein, X ₁ And X ₂ As defined above.

In some embodiments, the compound is selected from the group consisting of:

(2S,3S,4R) -1-O- (α -D-galactosyl) -2-N- ((S) -12-hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol;

(2S,3S,4R) -1-O- (α -D-galactosyl) -2-N- ((R) -12-hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol;

(2S,3S,4R) -1-O- (5-thio- α -D-galactosyl) -2-N- ((S) -12-hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol; and

(2S,3S,4R) -1-O- (5-thio-. alpha. -D-galactosyl) -2-N- ((R) -12-hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol.

In some embodiments, the present application is directed to a pharmaceutical composition comprising: alpha-galactosyl ceramide compound shown in general formula (I) or pharmaceutically acceptable salt or solvate thereof; and pharmaceutically acceptable auxiliary materials.

The pharmaceutical composition of the present application may be formulated into any form of preparations, such as capsules, tablets, aerosols, solutions, suspensions, dragees, syrups, emulsions, ointments, pastes, injections, powders, granules, pastes, sustained-release preparations, foams. The drug of the present application may be formulated into an oral administration formulation, a nasal administration formulation, a pulmonary administration formulation, an buccal formulation, a subcutaneous administration formulation, an intradermal administration formulation, a transdermal administration formulation, a parenteral administration formulation, a rectal administration formulation, a depot administration formulation, an intravenous administration formulation, an intraurethral administration formulation, an intramuscular administration formulation, an intranasal administration formulation, an ophthalmic administration formulation, an epidural administration formulation or a topical administration formulation according to the administration route.

The adjuvants described herein may be any pharmaceutically acceptable adjuvant, such as, but not limited to, solvents, propellants, solubilizers, solubilizing agents, emulsifiers, colorants, disintegrants, fillers, lubricants, wetting agents, tonicity adjusting agents, stabilizers, glidants, flavoring agents, preservatives, suspending agents, antioxidants, permeation enhancers, pH adjusting agents, surfactants, diluents, and the like. For other pharmaceutically acceptable pharmaceutical excipients that can be used, see for example the handbook of pharmaceutical excipients (4 th edition), monograph on r.c. ro, zheng folk translation, 2005, chemical industry press.

In one embodiment, the application relates to the use of an α -galactosylceramide compound represented by the above general formula (I) or a pharmaceutically acceptable salt or solvate thereof or the above pharmaceutical composition for the preparation of a medicament for inducing a Th1 selective immune response.

In one embodiment, the present application relates to the use of an α -galactosylceramide compound of the above general formula (I) or a pharmaceutically acceptable salt or solvate thereof or the above pharmaceutical composition for the preparation of one or more of the following medicaments: an antitumor agent such as an agent for treating or ameliorating melanoma, breast cancer, non-small cell lung cancer, liver cancer, stomach cancer, cervical cancer, colon cancer, leukemia, prostate cancer, brain cancer, skin cancer, bone cancer, lymph cancer, nasopharyngeal cancer, laryngeal cancer, esophageal cancer, duodenal cancer, small intestine cancer, large intestine cancer, pancreatic cancer, renal cancer, genital cancer and/or a thyroid cancer; antiviral drugs, such as drugs for treating or alleviating diseases associated with murine pox virus, influenza virus and/or hepatitis virus; antibacterial drugs; antituberculotic agents; and agents that modulate autoimmune activity.

The cytokine detection result of the invention shows that the alpha-GalCer analogue can induce the selective immune response of Th1 by increasing the yield of the Th1 cytokines or reducing the yield of the Th2 cytokines, and has stronger selective secretion inductivity of Th 1. The research on the anti-virus activity and the anti-cancer activity shows that the alpha-GalCer analogue has obviously better anti-virus and anti-cancer activity, and can be used as an anti-tumor medicament, an anti-virus medicament, an antibacterial medicament, an anti-tuberculosis medicament and a medicament for regulating the autoimmune activity.

In one embodiment, the present application relates to a method for preparing an α -galactosylceramide compound represented by the above general formula (I) or a pharmaceutically acceptable salt or solvate thereof, the method comprising the steps of:

(1) condensing a compound represented by formula (M) with a compound represented by formula (N) in the presence of a catalyst to produce a compound represented by formula (Q); wherein the catalyst is selected from one or more of N-iodosuccinimide (NIS), N-bromosuccinimide (NBS) and boron trifluoride diethyl etherate, trifluoromethanesulfonic acid, silicon trifluoromethanesulfonate and tert-butyldimethylsilyl trifluoromethanesulfonate, preferably the catalyst is selected from one or more of trimethylsilyl trifluoromethanesulfonate and NIS;

(2) removing the hydroxy protecting group from the compound of formula (Q), and optionally sulfating the removed hydroxy protecting group to produce an alpha-galactosylceramide compound of formula (I);

wherein, X ₁ And X ₂ And R ₁ -R ₉ As defined above;

y is-Br, -SCH (CH) ₃ ) ₂ -SPh, -OAc or-OC (NH) CCl ₃ (ii) a Z is-OH or-SH.

Preferably, the compounds of the formula (N) are prepared starting from commercial phytosphingosines.

The advantages of this route are: the reaction condition is mild, the operation is convenient, and the method can be used for industrial preparation. Synthetic routes can be used to prepare different types of α -GalCer analogs.

Examples

The present invention will be described in further detail with reference to examples, but the present invention is not limited to these examples. The experimental methods used in the following examples are conventional methods unless otherwise specified; reagents, materials, devices and the like used in the following examples are commercially available or can be prepared by those skilled in the art according to the ordinary skill in the art, unless otherwise specified.

Example 1: synthesis of alpha-GalCer-OH analogs

alpha-GalCer-12R-OH and alpha-GalCer-12S-OH were synthesized by the following scheme.

Reagents and conditions: (a) LHMDS, THF/HMPA 6/1, -78 ℃ to rt, 57%; (b) 80% AcOH aq.60 ℃; (c) TBDPSCl, pyr.91%; (d) BnOC (NH) Cl ₃ ,TMSOTf,DCM,84％；(e)HF-pyr.,DCM, rt,95％；(f)(i)Dess-Martin periodinane,DCM,rt；(ii)C ₁₃ H ₂₇ Ph ₃ P ⁺ Br ^- LHMDS, THF, -78 ℃ to rt, 54%, two steps; (g) LiOH. H ₂ O,THF/MeOH/H ₂ O(1:1:1)；1N HCl aq.,92％。

Reagents and conditions: (a) (i)12, EDCI, HOBt, Et ₃ N, DMF; (ii) TBDPSCl, pyr, rt; (iii) BzCl,66 percent, and three steps; (b)4bar H ₂ ,Pd(OH) ₂ /C,EtOAc,94％；(c)HF-pyr.,DCM,rt,95％；(d)TMSOTf,NIS,DCM,

Molecular sieves, N ₂ ,0℃,64％；(e)4bar H ₂ ,Pd(OH) ₂ MeOH/EtOAc (4: 1); (f) NaOMe, MeOH, 88%, two steps.

(R, Z) -11- (2, 2-dimethyl-1, 3-dioxan-4-yl) -10-undecenoic acid methyl ester (6a)

The phosphonium salt 5a (9.13g,20mmol) was dissolved in 140mL of anhydrous THF/HMPA (V/V:6/1) and the reaction cooled to-78 ℃. LHMDS (2.5M in THF,8ml, 20mmol) was added under nitrogen and the reaction stirred for 30min under these conditions. 4(2g, 10mmol) was dissolved in 2mL THF and slowly added to the reaction system of 5a, after stirring for 30min the reaction system slowly rose to 0 ℃. The reaction solution was quenched with saturated aqueous ammonium chloride solution. THF was then evaporated off and the aqueous phase was extracted with ethyl acetate (3X 50 mL). The organic solvent was evaporated under pressure and the resulting concentrate was isolated by column chromatography (EtOAc/petroleum ether: 1/15) to give 1.7g of the desired product 6a as a colorless oil in 57% yield.

(S, Z) -11- (2, 2-dimethyl-1, 3-dioxan-4-yl) -10-undecenoic acid methyl ester (6b)

The operation and the charge ratio refer to the synthesis method of the compound 6 a.

(R, Z) -11- (12, 13-dihydroxy) -10-tridecanoic acid methyl ester (7a)

6a (2g, 6.7mmol) was added to 20mL of 80% AcOH in water and heated at 60 deg.C, the starting material was detected by TLC and after completion of the reaction, it was distilled under reduced pressure, and the concentrate was separated by column chromatography (EtOAc/petroleum ether: 1/1) to give 1.61g of the title product 7a as a colorless oil in 93% yield.

(S, Z) -11- (12, 13-dihydroxy) -10-tridecanoic acid methyl ester (7b)

The operation and the charge ratio refer to the synthesis method of the compound 7 a.

(R, Z) -11- [13- (tert-butyldiphenylsilyl) oxy ] -12-hydroxy-10-tridecanoic acid methyl ester (8a)

Compound 7a (2g, 7.7mmol) was dissolved in 30mL pyridine, TBDPSCl (2.4mL, 9.3mmol) was added under ice bath, and the reaction was allowed to warm to room temperature for 12 h. The reaction was checked by TLC and starting material disappeared. The pyridine in the reaction was distilled off, and the resulting concentrate was subjected to column chromatography (EtOAc/petroleum ether: 1/15) to give 3.5g of 8a in 91% yield.

(S, Z) -11- [13- (tert-butyldiphenylsilyl) oxy ] -12-hydroxy-10-tridecanoic acid methyl ester (8b)

The operation and the charge ratio refer to the synthesis method of the compound 8 a.

(R, Z) -11- (12-benzyloxy) - [13- (tert-butyldiphenylsilyl) oxy ] -10-tridecanoic acid methyl ester (9a)

8a (1g, 2mmol) is dissolved in anhydrous 30mL dichloromethane, and 2mg 4A molecular sieve, BnO (CNH) CCl are added in sequence at 0 ℃ under the protection of nitrogen ₃ (0.5mL, 2.6mmol) and TMSOTf (18. mu.L, 1. mu. mol), then maintained at 0 ℃ for 3h and quenched by the addition of 3 drops of triethylamine. The reaction mixture was extracted with 10mL of saturated brine, and the organic phase was dried over anhydrous sodium sulfate. The distillation was carried out under pressure, and the concentrate was subjected to column chromatography (EtOAc/petroleum ether: 1/30) to give 9a (0.99g, 84%).

(S, Z) -11- (12-benzyloxy) - [13- (tert-butyldiphenylsilyl) oxy ] -10-tridecanoic acid methyl ester (9b)

The operation and the charge ratio refer to the synthesis method of the compound 9 a.

(R, Z) -11- (12-benzyloxy) -13-hydroxy-10-tridecanoic acid methyl ester (10a)

Compound 9a (1g, 1.7mmol) was dissolved in 15mL of dichloromethane and pyridine hydrofluoric acid solution (313. mu.L, 2.1mmol) was added in an ice bath. The reaction was carried out for about 5 h. And adding saturated sodium bicarbonate solution into the reaction solution for quenching. The reaction was then extracted with dichloromethane (2X 10 mL). The organic phase was separated and dried over anhydrous sodium sulfate. The concentrate obtained by distilling off the organic solvent was subjected to column chromatography (EtOAc/petroleum ether: 1/3) to obtain 10a (0.56g, 95%).

(S, Z) -11- (12-benzyloxy) -13-hydroxy-10-tridecanoic acid methyl ester (10b)

The operation and charge ratio were referenced to the synthesis of compound 10 a.

(S,10Z,13Z) -12-benzyloxy-hexacosanedienoic acid methyl ester (11a)

10a (1g, 2.9mmol) was dissolved in 10mL of dichloromethane and Dess-Martin oxidant (1.6g, 3.77mmol) was added at 0 ℃. The reaction was stirred at room temperature for 6h, then over 40mL Na ₂ S ₂ O ₃ The reaction was quenched with saturated aqueous solution, the solution was extracted with dichloromethane (3 х 30mL), the organic phase was dried over anhydrous sodium sulfate, and the organic solvent was evaporated to give the aldehyde. C is to be ₁₃ H ₂₇ Ph ₃ P ⁺ Br ^- (1.8g, 3.4mmol) was added to 30mL THF, LHMDS (2.5M in THF,1.4mL, 3.4mmol) was slowly added at-78 deg.C under nitrogen, the solution was reacted for 30min, and the aldehyde, previously prepared, was added and maintained at-78 deg.C for 1 h. Gradually raising the temperature to room temperature, and adding NH ₄ Cl 10mL, distilled under reduced pressure, 30mL of water was added to the crude product, and the aqueous phase was extracted with ethyl acetate (2 х 30 mL). The organic phase was dried over anhydrous sodium sulfate. Distillation under reduced pressure and separation of the crude product by column chromatography gave compound 11a (795mmg, 54%).

(R,10Z,13Z) -12-benzyloxy-hexacosanedienoic acid methyl ester (11b)

The operation and charge ratio were determined according to the synthesis method of compound 11 a.

(S,10Z,13Z) -12-benzyloxy-hexacosanedienoic acid (12a)

Compound 11a (600)mg, 1.2mmol) was added to 18mL THF/MeOH/H ₂ O (v/v/v:1/1/1), then adding LiOH ₂ In O (196mg, 4.7mmol), stir at room temperature for 12 h. The reaction solution was adjusted to pH 3 to 4 with 1N aqueous hydrochloric acid solution. The reaction solution was extracted with dichloromethane (3X 30 ml). The organic phase was washed once with saturated brine (40 ml). The organic phase was dried over anhydrous sodium sulfate. The organic solvent was distilled off under reduced pressure, and the resulting concentrate was subjected to column chromatography (EtOAc/petroleum ether: 1/2) to give the objective product 12a (537mg, 92%) as a colorless oil.

(R,10Z,13Z) -12-benzyloxy-hexacosanedienoic acid (12b)

The operation and charge ratio refer to the synthesis method of the compound 12 a.

(2S,3S,4R) -2-N- ((S, 10Z,13Z) -12-benzyloxy-hexacosanoylamino) -3, 4-di-O-benzoyl-1, 3, 4-octadecanetriol (13a)

To a solution of 12a (1g, 2mmol) in DMF was added HOBt (296mg, 2.2mmol) and EDCI (422mg, 2.2mmol) in that order. The reaction was stirred at room temperature for 30min, then phytosphingosine 2(635mg, 2mmol) and DIPEA (0.8ml, 4.6mmol) were added. The reaction was stirred at room temperature for about 12 h. TLC detects that the reaction is complete. The DMF was evaporated and the concentrate was added to 50mL of water and extracted with dichloromethane (3X 50 mL). The organic phase was dried over anhydrous sodium sulfate, then the crude product was dissolved in 30mL pyridine by distillation under reduced pressure, TBDPSCl (1.2mL, 4.8mmol) was added under ice bath, and the reaction was allowed to warm to room temperature for 12 h. The reaction was checked by TLC and starting material disappeared. Benzoyl chloride (1.8ml, 16mmol) was then added to the reaction and after 12h at rt the pyridine was evaporated off and the resulting concentrate was isolated by column chromatography (10% EtOAc in petroleum ether) to give compound 13a as a colourless oil (17g, 66%, three steps).

(2S,3S,4R) -2-N- ((R, 10Z,13Z) -12-benzyloxy-hexacosanoylamino) -3, 4-di-O-benzoyl-1, 3, 4-octadecanetriol (13b)

The operation and the charge ratio refer to the synthesis method of the compound 13 a.

(2S,3S,4R) -2-N- ((S) -12-Hydroxyhexacosanoylamino) -3, 4-di-O-benzoyl-1, 3, 4-octadecanetriol (15a)

13a (1g, 0.8mmol) was added to 20mL ethyl acetate, followed by Pd (OH) ₂ The catalyst was prepared as a/C (20 wt% dry basis on carbon, 50mg) and the hydrogenation reduced at 4 atm for about 12 h. Suction filtration, filter cake 10mL CHCl ₃ (4X 10mL) and the filtrate evaporated to dryness. The concentrated solution was dissolved in 15mL of dichloromethane, and pyridine hydrofluoric acid solution (676. mu.L, 2.44mmol) was added in an ice bath. The reaction was carried out for about 5 h. Adding a saturated solution of sodium bicarbonate into the reaction solution for quenching. The reaction was then extracted with dichloromethane (2X 10 mL). The organic phase was separated and dried over anhydrous sodium sulfate. The organic solvent was distilled off to give a concentrate, which was subjected to column chromatography (EtOAc/petroleum ether: 3/5) to give 15a (757mg, 95%) as a colorless oily compound.

¹ H NMR(400MHz，CDCl ₃ )δ8.05 (d，J＝7.4Hz，2H)，7.94(d，J＝7.4Hz，2H)，7.63(t，J＝7.5Hz， 1H)，7.51(dt，J＝12.8，7.5Hz，3H)，7.38(t，J＝7.7Hz，2H)， 6.42(d，J＝9.3Hz，1H)，5.41(dd，J＝9.6，2.4Hz，1H)，5.36(dt，J＝9.1，4.1Hz，1H)，4.38(t，J＝9.4Hz，1H)，3.69–3.54(m，3H)， 2.38–2.19(m，2H)，2.02(m，4H)，1.67(m，2H)，1.47–1.15 (m，64H)，0.87(t，J＝6.6Hz，6H)； ¹³ C NMR(101MHz，CDCl ₃ ) δ173.4，167.3，166.5，134.0，133.3，130.1，130.0，129.8，129.2，128.8， 128.5，74.1，73.9，72.2，61.7，50.0，37.7，37.6，36.9，32.1，29.9， 29.8，29.8，29.8，29.7，29.7，29.6，29.6，29.5，29.5，29.4，29.3，28.6， 27.4，26.0，25.8，25.8，25.7，22.8，14.3；HRMS(ESI)m/z calcd for C ₅₈ H ₉₃ O ₇ NNa[M+Na] ⁺ 938.6844，found 938.6851.

(2S,3S,4R) -2-N- ((R) -12-Hydroxyhexacosanoylamino) -3, 4-di-O-benzoyl-1, 3, 4-octadecanetriol (15b)

The operation and the charge ratio refer to the synthesis method of the compound 15 a.

(2S,3S,4R) -2-N- ((S) -12-Hydroxyhexacosanoylamino) -3, 4-di-O-benzoyl-1-O- (2,3,4, 6-tetra-O-benzyl-alpha-D-galactosyl) -1,3, 4-octadecanetriol (16a)

Under the protection of nitrogen, 15a (200mg, 0.21mmol) and 3-24(136mg, 0.23mmol) are dissolved in 10mL of dichloromethane, and N-iodosuccinimide (NIS, 57mg, 0.25mmol) and,

Molecular sieve powder, stirring at 0 deg.C for 15 min. TMSOTf (2. mu.L, 11. mu. mol) was added rapidly to the reaction. The reaction solution was continuously reacted at 0 ℃ for 5 hours. The reaction was completed by TLC detection, and then quenched by the addition of 10mL of saturated sodium thiosulfate solution and 1mL of saturated sodium bicarbonate solution, the reaction was extracted with dichloromethane (2X 15mL), and the organic phase was separated and dried over anhydrous sodium sulfate. Concentrated under reduced pressure and the resulting concentrate was subjected to column chromatography (EtOAc/petroleum ether: 1/5) to give 16a (199mg, 58%).

¹ H NMR(400MHz，CDCl ₃ )δ8.03 (d，J＝7.8Hz，2H)，7.93(d，J＝7.8Hz，2H)，7.59(t，J＝7.4Hz， 1H)，7.51(t，J＝7.4Hz，1H)，7.45(t，J＝7.7Hz，2H)，7.36(t， J＝7.7Hz，2H)，7.29–7.23(m，3H)，7.20–7.08(m，6H)，6.85 (d，J＝8.3Hz，2H)，6.79(dd，J＝15.7，8.3Hz，4H)，6.70(d，J ＝8.3Hz，2H)，5.70(d，J＝9.6Hz，1H)，5.40(t，J＝6.6Hz，1H)， 4.77(d，J＝11.3Hz，1H)，4.68(d，J＝3.6Hz，1H)，4.64(d，J＝11.2 Hz，1H)，4.56(d，J＝11.3Hz，4H)，4.45(t，J＝11.2Hz，2H)， 4.32(d，J＝11.8Hz，1H)，4.01(t，J＝6.3Hz，1H)，3.98–3.88(m， 2H)，3.82–3.72(m，13H)，3.63–3.52(m，2H)，3.47–3.39(m， 1H)，3.24(dd，J＝9.4，5.4Hz，1H)，2.16(t，J＝7.6Hz，2H)， 1.89(m，2H)，1.62(m，2H)，1.48–1.13(m，70H)，0.88(t，J＝ 6.4Hz，6H)； ¹³ C NMR(101MHz，CDCl ₃ )δ173.3，166.2，165.4，159.3，159.3，159.3，159.2，133.3，133.0，131.1，130.7，130.6，130.4，130.1， 130.0，129.9，129.8，129.7，129.2，128.7，128.4，113.9，113.7，100.3， 78.5，76.4，74.6，74.3，74.0，73.2，73.0，73.0，72.3，72.2，70.4，70.1， 69.0，55.4，48.7，37.7，37.7，36.8，32.1，29.9，29.8，29.8，29.7，29.7， 29.6，29.5，28.5，25.8，25.8，22.8，14.3；MALDI-TOF MS m/z calcd for C ₉₂ H ₁₃₁ NO ₁₂ SiNa[M+Na] ⁺ 1465.0，found 1464.9.

(2S,3S,4R) -2-N- ((R) -12-Hydroxyhexacosanoylamino) -3, 4-di-O-benzoyl-1-O- (2,3,4, 6-tetra-O-benzyl-alpha-D-galactosyl) -1,3, 4-octadecanetriol (16b)

The operation and the charge ratio refer to the synthesis method of the compound 16 a.

(2S,3S,4R) -1-O- (. alpha. -D-galactosyl) -2-N- ((S) -12-Hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol (. alpha. -GalCer-12S-OH)

3-36a (200mg, 0.14mmol) was added to 4mL of methanol and 1mL of ethyl acetate, followed by Pd (OH) ₂ C (20 wt.% dry basis on carbon, 20mg) as catalyst, and reduced by hydrogenation at 4 atm for about 24 h. Suction filtration is carried out, a filter cake is washed by 10mL of dichloromethane, filtrate is evaporated to dryness, the obtained concentrate is added into 5mL of methanol, and 1M sodium methoxide methanol solution is added to adjust the pH of a reaction system to 9-10. The reaction was stirred at room temperature until the reactants were completely reacted, and then IR-120 (H) was used ⁺ ) Neutralizing with cationic resin, vacuum filtering, washing filter cake with dichloromethane, and mixing organic phases. The organic phase is evaporated to dryness and the concentrate is subjected to column chromatography (MeOH/CH) ₂ Cl ₂ 1/5) to obtain the compound α -GalCer-12S-OH (107mg, 88%).

¹ H NMR(400MHz，MeOD)δ4.81 (d，J＝3.8Hz，1H)，4.11(q，J＝4.7Hz，1H)，3.84(d，J＝3.3Hz， 1H)，3.80(dd，J＝10.8，4.6Hz，1H)，3.75-3.56(m，6H)，3.49- 3.41(m，3H)，2.12(t，J＝7.6Hz，2H)，1.63-1.43(m，4H)，1.36 -1.12(m，66H)，0.79(t，J＝6.7Hz，6H). ¹³ C NMR(101MHz， MeOD)δ174.8，100.0，75.1，72.3，72.0，71.1，70.6，70.0，69.2，62.1， 50.7，37.5，36.7，32.9，32.2，30.0，30.0，30.0，29.9，29.9，29.8，29.7， 29.6，29.6，26.1，25.9，25.9，22.9，14.2；HRMS(ESI)m/z calcd for C ₅₀ H ₉₉ NO ₁₀ Na[M+Na] ⁺ 896.7161，found 896.7166.

(2S,3S,4R) -1-O- (. alpha. -D-galactosyl) -2-N- ((R) -12-Hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol (. alpha. -GalCer-12R-OH)

The operation and the feeding ratio refer to the synthesis method of the compound alpha-GalCer-12S-OH.

¹ H NMR(400MHz,CDCl ₃ /CD ₃ OD:5/1) δ4.91(d,J＝3.9Hz,1H),4.20(q,J＝4.6Hz,1H),3.94(d,J＝3.3Hz,1H), 3.89(dd,J＝10.7,4.7Hz,1H),3.83–3.66(m,6H),3.54(d,J＝5.2Hz,3H), 2.21(t,J＝7.7Hz,2H),1.62(td,J＝24.0,21.5,13.4Hz,5H),1.28(d,J＝ 8.4Hz,89H),0.88(t,J＝6.7Hz,8H)； ¹³ C NMR(101MHz,CD ₃ OD)δ174.8, 100.0,75.1,72.3,72.0,71.1,70.6,70.0,69.2,62.1,50.7,37.5,36.7,32.9, 32.2,30.0,30.0,30.0,29.9,29.9,29.8,29.7,29.6,29.6,26.1,25.9,25.9, 22.9,14.2；HRMS(ESI)m/z calcd for C ₅₀ H ₉₉ NO ₁₀ Na[M+Na] ⁺ 896.7161, found 896.7164.

Example 2: synthesis of 5-Thio-alpha-GalCer-OH analogs

5-Thio- α -GalCer-12S-OH and 5-Thio- α -GalCer-12R-OH were synthesized by the following schemes. Unless otherwise indicated, the starting materials used in the examples, such as galacto-sulphur starting material 18, were obtained from Du subjects.

Reagents and conditions: (a) the amount of TMSOTf, DCM,

molecular sieves, N ₂ 0 ℃ for 2h, 56%; (b) NaOMe, MeOH, rt,6h, quant

(2S,3S,4R) -2-N- ((S) -12-Hydroxyhexacosanoylamino) -3, 4-di-O-benzoyl-1-O- (2,3,4, 6-tetra-O-acetyl-5-thio-. alpha. -D-galactosyl) -1,3, 4-octadecanetriol (19a)

Under the protection of nitrogen, 15a (100mg, 0.11mmol) and freshly dried

Molecular sieves (40mg) were added to 1.5mL of dichloromethane and the reaction was stirred at 0 ℃ for 30 min. TMSOTf (2. mu.L, 11. mu. mol) was added rapidly, and then 18(54mg,0.11mmol) was dissolved in 0.5mL of dichloromethane, and slowly dropped into the reaction system under nitrogen protection, followed by reaction at 0 ℃ for 4 hours. 2 drops of triethylamine are added, followed by 15mL of dichloromethane and saturated NaHCO ₃ The aqueous solution was extracted once, and the saturated aqueous NaCl solution was extracted once. The organic phase was dried over anhydrous sodium sulfate. Concentrated under reduced pressure and the concentrate was subjected to column chromatography (EtOAc/petroleum ether: 1/4) to give compound 19a (75mg, 56%).

(2S,3S,4R) -2-N- ((R) -12-Hydroxyhexacosanoylamino) -3, 4-di-O-benzoyl-1-O- (2,3,4, 6-tetra-O-acetyl-5-thio-. alpha. -D-galactosyl) -1,3, 4-octadecanetriol (19b)

The operation and the charge ratio refer to the synthesis method of the compound 19 a.

(2S,3S,4R) -1-O- (5-thio-. alpha. -D-galactosyl) -2-N- ((S) -12-Hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol (5-thio-. alpha. -GalCer-12S-OH)

Adding compound 19a (50mg, 0.04mmol) into 4mL of methanol, adjusting pH of the reaction solution to 9-10 with 1M sodium methoxide solution in methanol, reacting for 12H, detecting the reaction completion with TCL, and detecting with IR-120 (H) ⁺ ) The cationic resin is adjusted to neutrality. The filtrate was evaporated to dryness and separated by column chromatography to obtain 35mg of a white solid target product, 5-thio- α -GalCer-12S-OH.

¹ H NMR(400MHz,Pyridine-d ₅ )δ8.42 (d,J＝8.7Hz,1H),5.36(d,J＝2.8Hz,1H),5.30(dt,J＝8.9,4.5Hz,1H), 4.96(s,1H),4.85(d,J＝9.7Hz,1H),4.77(dd,J＝10.7,5.5Hz,1H),4.46– 4.25(m,5H),4.21(dd,J＝10.7,5.5Hz,1H),3.96(t,J＝6.7Hz,1H),3.92– 3.83(m,1H),2.43(t,J＝7.4Hz,2H),2.24–2.35(m,1H),1.99–1.51(m, 13H),1.49–1.13(m,56H),0.94–0.78(m,6H)； ¹³ C NMR(101MHz, Pyridine-d ₅ )δ173.0,85.2,76.8,72.8,72.4,72.3,71.6,70.9,69.4,62.3,50.7, 45.8,38.5,36.7,34.3,32.1,30.3,30.2,30.1,29.9,29.9,29.8,29.7,29.6, 29.5,26.4,26.3,22.9,14.2；HRMS(ESI)m/z calcd for C ₅₀ H ₉₉ NO ₉ SNa [M+Na] ⁺ 912.6933,found 912.6931.

(2S,3S,4R) -1-O- (5-thio-. alpha. -D-galactosyl) -2-N- ((R) -12-hydroxyhexadecanoylamino) -1,3, 4-octadecanetriol (5-thio-. alpha. -GalCer-12R-OH)

The operation and the feeding ratio refer to the synthesis method of the compound 5-sulfur-alpha-GalCer-12S-OH.

¹ H NMR(400MHz,Pyridine-d ₅ )δ8.45 (d,J＝8.6Hz,1H),5.35(d,J＝3.2Hz,1H),5.32–5.26(m,1H),4.96(s, 1H),4.85(d,J＝8.9Hz,1H),4.81–4.73(m,1H),4.45–4.25(m,6H),4.21 (dd,J＝10.7,5.6Hz,1H),3.96(t,J＝6.7Hz,1H),3.89(d,J＝7.3Hz,1H), 2.43(t,J＝7.5Hz,2H),2.34–2.23(m,1H),1.97–1.53(m,13H),1.50– 1.15(m,56H),0.86(t,J＝6.7Hz,6H)； ¹³ C NMR(101MHz,Pyridine-d ₅ )δ 173.0,85.2,76.7,72.7,72.3,72.2,71.6,70.9,69.4,62.2,50.7,50.6,45.8, 38.5,36.7,36.6,34.3,32.0,30.3,30.2,30.1,29.9,29.8,29.8,29.7,29.7, 29.5,26.4,26.3,22.9,14.2；HRMS(ESI)m/z calcd for C ₅₀ H ₉₉ NO ₉ SNa [M+Na] ⁺ 912.6933,found 912.6935.

Example 3: induction of Th1 Selective immune response Studies

1. Test material

C57BL/6 mice (six weeks old, weight: 20-25 g), Beijing Witongliwa laboratory animal technology, Inc., IFN-. gamma.and IL-4ELISA kits, purchased from Sigma-Aldrich.

And (3) detecting the medicine: the positive control compound α -GalCer, and the test samples α -GalCer-12S-OH (also called GC _12S, HP-1) and 5-Thio- α -GalCer-12S-OH (also called GCs _12S, HP-4).

2. Test method

Mice were injected intravenously with PBS (blank control), α -GalCer (KRN7000 positive control), HP-1 or HP-4, 4 nmol/mouse, and blood was taken at 0h (pre-dose), 3h, 6h, 9h, 12h, 24h and 48h time points, respectively, to determine IFN-. gamma.and IL-4 concentrations. The selective propensity of a compound to induce Th1/Th2 of a relevant cytokine can be expressed as the ratio of the maximum secretion (i.e., peak) of IFN- γ/IL-4 and/or the area under the line (AUC) ratio of the IFN- γ/IL-4 secretion curve, with a greater ratio indicating a greater compound activity to induce selectivity for Th1/Th2 of the relevant cytokine.

FIG. 3 results show that HP-1 and HP-4 induced a higher concentration of the Th1 type cytokine IFN-. gamma.than alpha-GalCer (KRN 7000); the peak value ratios of IFN-gamma and IL-4 generated by induction of alpha-GalCer (KRN7000), HP-1 and HP-4 are 2.1, 2.2 and 11.6 in sequence; the area under line (AUC) ratios were 10.32, 15.44, and 79.63, respectively. Thus, compared to α -GalCer, the α -GalCer analogs HP-1 and HP-4, which have monohydroxy substitution at specific positions of the α -GalCer amide chain, have significantly stronger activity to induce a Th1 selective immune response. In addition, unexpectedly, 5-S-alpha-GalCer-OH (HP-4) has significantly stronger Th1/Th2 selective induction secretion activity, even stronger than the alpha-GalCer analogue with double hydroxyl substitution at C-12 and 13 of the amide chain (the AUC ratio of the area under the line of IFN-gamma/IL-4 secretion curve is 19.46: 1).

Example 4: study of antiviral Activity

19C 57BL/6 mice, 6 weeks old, were divided into four groups and were administered intravenously (4 nmol/mouse in 200uL PBS solution) with PBS (blank control, 4 mice), α -GalCer (KRN7000 group, 5 mice), HP-1 group (5 mice), or HP-4 group (5 mice) in PBS solution, respectively, and 3 days later were inoculated with the varicella virus. The survival monitoring results are shown in fig. 4.

As a result of activity test, the α -GalCer derivatives of HP-1 group in which hydroxyl group was introduced into the amide chain or HP-4 group in which galactose was simultaneously replaced with 5-thiogalactose significantly prolonged the survival time of mice with murine pox disease compared to the blank control group and α -GalCer (KRN7000) group, and thus their anti-murine pox virus activity was significantly stronger than KRN 7000.

Example 5: study of anti-melanoma Activity

16 mice, C57BL/6, 6 weeks old, were randomized into four groups (4/group) and administered intravenously (4 nmol/mouse, 200uL PBS solution in solution) with PBS solution (blank control group), α -GalCer (KRN7000 group), HP-1 group, or HP-4 group, 2 x 10 after 3 days ⁵ B16-F10 mouse melanoma cells. Three weeks later, the mice were sacrificed, and the lungs were taken to observe the growth and metastasis of melanoma, and the results are shown in FIG. 5, wherein the number and volume of melanoma in KRN7000, HP-1 and HP-4 groups were significantly smaller than those in the blank control group; the number and volume of melanoma of HP-1 group and HP-4 group are significantly less than KRN7000 group, so HP-1 and HP-4 can inhibit growth and metastasis of melanoma, and have anti-melanoma effect, and activity thereof is significantly stronger than KRN 7000.

Example 6: study of anti-Breast cancer Activity

20 6 week old immunodeficiency female NSG mice were randomly divided into four groups (5/group, blank control group, KRN7000 group, HP-1 group and HP-4 group), and the mice were injected into the mammary gland with 2 x 10 ⁶ Human breast cancer cell MDA-MB-231/Luc/hCD1 d. The survival was observed in four groups, i.v. (4 nmol/mouse, in 200uL PBS solution), PBS solution (blank control group), α -GalCer and human iNKT cells (KRN7000 group), α -GalCer-12S-OH and human iNKT cells (HP-1 group), or 5-Thio- α -GalCer-12S-OH and human iNKT cells (HP-4 group), at 3 days and 10 days after the tumor incorporation, respectively. As shown in FIG. 6, the survival rate of mice in KRN7000, HP-1 and HP-4 groups was significantly higher than that of the blank control group, and HP-4 and HP-1 groups were higher than that of KRN7000, which indicates that HP-1 and HP-4 had significantly stronger anti-breast cancer activity compared to KRN 7000.

Therefore, the alpha-GalCer analogue of the invention has remarkably excellent activity of inducing Th1 selective immune response and antitumor and antiviral activity.

The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention. It should be noted that, in the embodiment, the technical features described in the above embodiment can be combined in any suitable way without exception, and in order to avoid unnecessary repetition, the present invention does not need to describe any combination of the features. In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.

Claims (9)

1. An α -galactosylceramide compound represented by the general formula (I) or a pharmaceutically acceptable salt or solvate thereof:

wherein, X ₁ And X ₂ Each is independently selected from O or S;

R ₁ 、R ₂ and R ₃ Either one of them is hydroxyl or sulfate, the other two are both H; and is

R ₄ 、R ₅ 、R ₆ 、R ₇ 、R ₈ And R ₉ Each independently selected from hydroxyl or sulfate.

2. The α -galactosylceramide compound represented by the general formula (I) or a pharmaceutically acceptable salt or solvate thereof according to claim 1, wherein the compound has a structure represented by the following general formula (II):

wherein, X ₁ And X ₂ And R ₁ -R ₉ As defined in claim 1.

3. The α -galactosylceramide compound represented by the general formula (I) or a pharmaceutically acceptable salt or solvate thereof according to claim 1 or 2, wherein the compound has a structure represented by the following general formula (III):

wherein, X ₁ And X ₂ Each is independently selected from O or S; and is

R ₁ 、R ₂ And R ₃ Either one of them is a hydroxyl group or a sulfate group, and the remaining two are both H.

4. The α -galactosylceramide compound represented by the general formula (I) according to any one of claims 1 to 3, or a pharmaceutically acceptable salt or solvate thereof, wherein R is ₁ 、R ₂ And R ₃ One of the following three situations A, B and C is satisfied:

(A)R ₁ is hydroxy or sulfuric acid radical, and R ₂ And R ₃ Is H;

(B)R ₂ is hydroxy or sulfuric acid radical, and R ₁ And R ₃ Is H; and

(C)R ₃ is hydroxy or sulfuric acid radical, and R ₁ And R ₂ Is H;

wherein R is a hydroxyl group ₁ 、R ₂ And R ₃ The chirality of the carbon on which either is located is R or S.

5. The α -galactosylceramide compound represented by the general formula (I) or a pharmaceutically acceptable salt or solvate thereof according to any one of claims 1 to 4, which has a structure represented by the following general formula (IV) or general formula (V):

wherein, X ₁ And X ₂ As defined in claim 1;

preferably, the compound is selected from the group consisting of:

(2S,3S,4R) -1-O- (α -D-galactosyl) -2-N- ((S) -12-hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol;

(2S,3S,4R) -1-O- (α -D-galactosyl) -2-N- ((R) -12-hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol;

(2S,3S,4R) -1-O- (5-thio- α -D-galactosyl) -2-N- ((S) -12-hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol; and

(2S,3S,4R) -1-O- (5-thio-. alpha. -D-galactosyl) -2-N- ((R) -12-hydroxyhexacosanoylamino) -1,3, 4-octadecanetriol.

6. A pharmaceutical composition comprising: an α -galactosylceramide compound represented by the general formula (I) according to any one of claims 1 to 5, or a pharmaceutically acceptable salt or solvate thereof; and pharmaceutically acceptable adjuvants; preferably, the pharmaceutically acceptable adjuvant is selected from solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, antioxidants, permeation enhancers, pH regulators, surfactants or diluents.

7. Use of an α -galactosylceramide compound of the general formula (I) according to any one of claims 1 to 5 or a pharmaceutically acceptable salt or solvate thereof or a pharmaceutical composition according to claim 7 for the preparation of a medicament for inducing a Th1 selective immune response.

8. Use of an α -galactosylceramide compound of the general formula (I) according to any of claims 1 to 5 or a pharmaceutically acceptable salt or solvate thereof or a pharmaceutical composition according to claim 7 for the preparation of one or more of the following drugs: an antitumor agent such as an agent for treating or ameliorating melanoma, breast cancer, non-small cell lung cancer, liver cancer, stomach cancer, cervical cancer, colon cancer, leukemia, prostate cancer, brain cancer, skin cancer, bone cancer, lymph cancer, nasopharyngeal cancer, laryngeal cancer, esophageal cancer, duodenal cancer, small intestine cancer, large intestine cancer, pancreatic cancer, renal cancer, genital cancer and/or thyroid cancer; antiviral drugs, such as drugs for treating or alleviating diseases associated with murine pox virus, influenza virus and/or hepatitis virus; antibacterial drugs; antituberculotic agents; and agents that modulate autoimmune activity.

9. A process for the preparation of α -galactosylceramide compounds of the general formula (I) or pharmaceutically acceptable salts or solvates thereof as claimed in any of claims 1 to 5, said process comprising the steps of:

(1) condensing a compound represented by formula (M) with a compound represented by formula (N) in the presence of a catalyst to produce a compound represented by formula (Q); wherein the catalyst is selected from one or more of N-iodosuccinimide (NIS), N-bromosuccinimide (NBS), boron trifluoride diethyl etherate, trifluoromethanesulfonic acid, silicon triflate and tert-butyldimethylsilyl trifluoromethanesulfonate, preferably the catalyst is selected from one or more of trimethylsilyl triflate and N-bromosuccinimide (NBS);

(2) removing the hydroxy protecting group from the compound represented by formula (Q), and optionally sulfating the removed hydroxy group to produce an alpha-galactosylceramide compound represented by general formula (I);

wherein, X ₁ And X ₂ And R ₁ -R ₉ As set forth in the corresponding claimsDefining;

y is-Br, -SCH (CH) ₃ ) ₂ -SPh, -OAc or-OC (NH) CCl ₃ (ii) a Z is-OH or-SH.

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