JP4224429B2 - Sulfoquinovosyl acylglycerol - Google Patents

Description

  The present invention relates to a novel sulfoquinovosyl acylglycerol, and more particularly to a β-form sulfoquinovosyl acylglycerol.

  In the sulfoquinovosylacylglycerol derivative, the hydroxyl group of the 6-position carbon of D-glucose (hereinafter, the n-position carbon of the sugar is also referred to as “Cn carbon”) is substituted with a sulfo group, and the hydroxyl group of the C1 carbon is substituted with glycerol 6-deoxy-6-sulfo-D-glycopyranosylglycerol is used as a basic skeleton, and has a structure in which the hydroxyl group of the glycerol moiety is ester-linked with a fatty acid. There are many derivatives of sulfoquinovosyl acylglycerol derivatives depending on the type of fatty acid ester-bonded to glycerol. Among these sulfoquinovosyl acylglycerol derivatives, those having physiological activity expected to be applied to medicine are known.

  For example, in Ota et al., Non-Patent Document 1, a specific sulfoquinovosyl acylglycerol derivative obtained from red alga Suginori exhibits higher biological DNA synthase α and β inhibitory activity and HIV-derived reverse transcriptase inhibitory activity. It is described. However, Ota et al., Non-Patent Document 1, does not describe the anticancer activity.

  Non-patent document 2 of Suijin et al. Shows that a specific sulfoquinovosyl acylglycerol derivative obtained from a fern plant exhibits inhibitory activity on calf DNA synthase α-type and rat DNA synthase β-type. It is described that it does not affect the HIV-derived reverse transcriptase inhibitory activity. However, there is no description of the anticancer activity in Non-Patent Document 2 of Suijin.

On the other hand, Non-Patent Document 3 by Sahara et al. Describes that sulfoquinovosyl acylglycerol derivatives obtained from sea urchin components show anticancer activity in vivo and in vitro. However, the sulfoquinovosyl acylglycerol derivative found by Sawara et al. Is a mixture of multiple types of sulfoquinovosyl acylglycerol derivatives in which the acyl residues of fatty acids that are ester-linked to glycerol are different from each other. The single action of has not been clarified.
Chemical & Pharmaceutical Bulletin, 46 (4), (1998) Biochemical Pharmacology, 55, 537-541, (1998) British Journal of Cancer, 75 (3), 324-332, (1997)

  As described above, some sulfoquinovosyl acylglycerol derivatives are known to have physiological activity expected to be applied to pharmaceuticals, but those that show significant anticancer activity alone have not yet been found. Absent.

  The present inventors have found a compound having anticancer activity among sulfoquinovosyl acylglycerol derivatives. Among them, a compound in which the bond at the C1 carbon of glucose is a β bond is a novel compound.

  Accordingly, an object of the present invention is to provide such a novel sulfoquinovosyl acylglycerol derivative.

（式中、Ｒ₁₀₁は、飽和高級脂肪酸のアシル残基を表し、Ｒ₁₀₂は、水素原子又は飽和高級脂肪酸のアシル残基を表す。）により表される化合物およびその薬学的に許容される塩には制癌活性があることを見出した。そして、この一般式（１）により表される化合物のうち、グルコースのＣ１炭素における結合がβ結合であるものは、新規な化合物であり、本発明は、この新規なβ体のスルホキノボシルアシルグリセロール誘導体（以下、「本発明のβ誘導体」ともいう。）、すなわち次の一般式（２）：

（式中、Ｒ₁₀₁は、飽和高級脂肪酸のアシル残基を表し、Ｒ₁₀₂は、水素原子又は飽和高級脂肪酸のアシル残基を表す。）の構造のスルホン酸基が塩の形態にあるβ誘導体を提供する。 As a result of intensive studies, the present inventors have found that the following general formula (1):

(Wherein R ₁₀₁ represents an acyl residue of a saturated higher fatty acid, and R ₁₀₂ represents a hydrogen atom or an acyl residue of a saturated higher fatty acid) and pharmaceutically acceptable salts thereof Was found to have anticancer activity. Among the compounds represented by the general formula (1), those in which the bond at the C1 carbon of glucose is a β bond is a novel compound, and the present invention is a novel β-form sulfoquinovosyl. Acylglycerol derivatives (hereinafter also referred to as “β derivatives of the present invention”), that is, the following general formula (2):

(Wherein R ₁₀₁ represents an acyl residue of a saturated higher fatty acid, and R ₁₀₂ represents a hydrogen atom or an acyl residue of a saturated higher fatty acid) β derivative having a salt form of a sulfonic acid group I will provide a.

  The sulfoquinovosyl acylglycerol derivative represented by the general formula (1) including the β derivative represented by the general formula (2) has a significant anticancer activity. Such an anticancer agent containing at least one selected from the group consisting of sulfoquinovosylacylglycerol derivatives and pharmaceutically acceptable salts thereof as an active ingredient is highly expected to be used as a pharmaceutical product. .

  The sulfoquinovosyl acylglycerol derivative represented by the general formula (1) has an inhibitory action on the DNA synthase α-type (Assay 1 below). It is known that DNA synthase includes β type, γ type, δ type and ε type in addition to this α type. Of these DNA synthetases, the δ type and the ε type are considered to be in a biochemical type with the α type. Here, the biochemical type refers to having the following common enzyme functions. (I) Presence / absence of sensitivity to a specific compound. For example, these three types of DNA synthases have sensitivity to N-ethylmaleimide and butylphenyl-dGTP, but not to dideoxy TTP (ddTTP). (Ii) Fidelity: High accuracy of DNA synthesis for template DNA. (Iii) Reaction field: These three types of DNA synthases are both directly involved in DNA replication linked to cell division.

  DNA synthase α type (including δ type and ε type as biochemical types) is generally considered to control DNA synthesis according to the cell cycle. Therefore, the compound represented by the general formula (1) having an inhibitory activity against DNA synthase α-type (including δ-type and ε-type as a biochemical type) is a cancer in which cell proliferation occurs continuously and rapidly. It can be considered that it can have a growth-inhibiting ability for cells. The present inventors consider that the compound represented by the general formula (1) has an inhibitory activity on δ-type and ε-type DNA synthetases in addition to the α-type DNA synthetase.

  First, the anticancer agent (hereinafter also referred to as “the anticancer agent”) will be described in detail.

（式中、Ｒ₁₀₁は、飽和高級脂肪酸のアシル残基を表し、Ｒ₁₀₂は、水素原子又は飽和高級脂肪酸のアシル残基を表す。）により表される化合物およびその薬学的に許容される塩からなる群から選択される少なくとも１種を有効成分として含有する。 The anticancer agent found by the present inventors has the following general formula (1):

(Wherein R ₁₀₁ represents an acyl residue of a saturated higher fatty acid, and R ₁₀₂ represents a hydrogen atom or an acyl residue of a saturated higher fatty acid) and pharmaceutically acceptable salts thereof At least one selected from the group consisting of:

In the general formula (1), R ₁₀₁ represents an acyl residue of a saturated higher fatty acid. The fatty acid that provides the acyl residue of the saturated higher fatty acid represented by R ₁₀₁ includes a linear or branched saturated higher fatty acid. R ₁₀₁ is preferably an acyl residue of a linear saturated higher fatty acid, more preferably CH ₃ (CH ₂ ) _n CO— (n is 12), particularly from the viewpoint of anticancer activity against colorectal cancer and gastric cancer. It is a group represented by an integer of ˜24 (preferably an even number of 12 to 24).

In the general formula (1), _R102 represents a hydrogen atom or an acyl residue of a saturated higher fatty acid. Fatty acids that provide acyl residues of saturated higher fatty acids include linear or branched saturated higher fatty acids. In particular, from the viewpoint of antitumor activity against colorectal cancer and gastric cancer, R ₁₀₂ is preferably a hydrogen atom, but when R ₁₀₁ is CH ₃ (CH ₂ ) ₁₂ CO—, R ₁₀₂ is CH ₃ (CH ₂ ) Even ₁₂ CO- has exceptional anticancer activity.

  The bond between the sulfo-substituted glucose and glyceride of the compound represented by the general formula (1) may be an α bond or a β bond.

Of the compounds represented by the general formula (1), those preferable from the viewpoint of anticancer activity against colon cancer and stomach cancer are summarized in Table 1 below.

  Of the above compounds SQAG1 to SQAG14, SQAG1, SQAG2, SQAG4, SQAG6, SQAG8, SQAG11, SQAG12, SQAG13 and SQAG14 are preferred from the viewpoint of anticancer activity against gastric cancer or colon cancer.

  As described above, the anticancer agent contains at least one selected from the group consisting of the compound represented by the general formula (1) and a pharmaceutically acceptable salt thereof as an active ingredient.

  Pharmaceutically acceptable salts that can be used in the present anticancer agent include, but are not limited to, monovalent cation salts such as sodium and potassium. Hereinafter, the compounds of the group consisting of the compound of the general formula (1) and pharmaceutically acceptable salts thereof are also referred to as “the present anticancer active substance”.

  This anticancer active substance can be administered, for example, orally or parenterally. The anticancer active substance can be made into a pharmaceutical preparation by combining with an appropriate pharmaceutically acceptable excipient or diluent depending on the administration route.

  Dosage forms suitable for oral administration include those in a solid, semi-solid, liquid or gas state. Specifically, tablets, capsules, powders, granules, solutions, suspensions, Examples include syrups and elixirs, but are not limited thereto.

  In order to formulate the anticancer active substance into tablets, capsules, powders, granules, solutions, suspensions, etc., using a method known per se, the anticancer active substance is bound to a binder, It can be carried out by mixing with a tablet disintegrating agent, a lubricant and the like, and further mixing with a diluent, a buffering agent, a wetting agent, a preservative, a flavoring agent and the like, if necessary. For example, the binder includes crystalline cellulose, cellulose derivatives, corn starch, gelatin, etc., the tablet disintegrating agent includes corn starch, potato starch, sodium carboxymethyl cellulose, etc., and the lubricant includes talc, magnesium stearate, etc. In addition, conventionally used additives such as lactose and mannitol can be used.

  The anticancer active substance is in the form of a liquid, fine powder, together with a gas or liquid propellant, or optionally with a known auxiliary agent such as an infiltrating agent, in an aerosol container or nebulizer. Such a non-pressurized container can be filled and administered in the form of an aerosol or an inhalant. As the propellant, a pressurized gas such as dichlorofluoromethane, propane, or nitrogen can be used.

  When the anticancer agent is administered parenterally, it can be administered, for example, by rectal administration or injection.

  For rectal administration, for example, it can be administered as a suppository. Suppositories are molded by mixing the anticancer active substance with excipients such as cocoa butter, carbon wax, polyethylene glycol that melts at body temperature but solidifies at room temperature, in a manner known per se. Can be formulated.

  Administration by injection can be administered subcutaneously, intradermally, intravenously, intramuscularly or the like. In these injectable preparations, the anticancer active substance is dissolved in an aqueous or non-aqueous solvent such as vegetable oil, synthetic resin acid glyceride, higher fatty acid ester or propylene glycol by a method known per se. Suspended, emulsified and optionally formulated with conventionally used additives such as solubilizers, osmotic pressure regulators, emulsifiers, stabilizers and preservatives.

  In order to make this anticancer active substance into a solution, suspension, syrup, elixir or the like, a pharmaceutically acceptable solvent such as sterile water for injection or normal physiological saline can be used.

  This anticancer active substance can also be combined with a compound having other pharmaceutically acceptable activity to form a pharmaceutical preparation.

  The dose of the anticancer agent can be appropriately set and adjusted according to the administration form, administration route, the degree and stage of the target disease, and the like. For example, when administered orally, the anticancer active substance is 1 to 10 mg / kg body weight / day, and when administered as an injection, the anticancer active substance is 1 to 5 mg / kg body weight / day. In the case of rectal administration, the anticancer active substance can be set to 1 to 5 mg / kg body weight / day, but is not limited thereto.

  Cancers to which the anticancer agent can be effective include those having properties as malignant tumors, for example, mammalian adenocarcinoma including human, epithelial cancer, sarcoma, glioma, melanoma, lymphoma, leukemia There is.

  The compound represented by the general formula (1) can be efficiently produced by using pyranosides represented by the following general formula (A) and general formula (B) as intermediates. The pyranosides represented by these general formulas (A) and (B) are novel compounds.

  The pyranoside (1-O- (2-propenyl) -6-O-sulfonylpyranoside) represented by the general formula (A) will be described in detail.

により表されるピラノシドを構成する糖骨格であるピラノースには、α−Ｄ−グルコース、β−Ｄ−グルコースが含まれる。これらの糖骨格は、舟形、いす型のいずれの配置をもとり得る。しかしながら、いす型のもののほうが、安定性の観点から好ましい。 Formula (A):

The pyranose that is a sugar skeleton constituting the pyranoside represented by the formula includes α-D-glucose and β-D-glucose. These sugar skeletons can take either a boat shape or a chair shape. However, the chair type is preferable from the viewpoint of stability.

In the general formula (A), the 2-propenyl group bonded to the C1 carbon may be an α bond or a β bond.
In the general formula (A), R ¹ , R ² and R ³ each independently represent an alkyl group or a substituted silyl group. R ¹ , R ² and R ³ may be the same or different from each other. However, these three substituents are preferably the same as each other from the viewpoint of ease of production.

The alkyl group represented by R ¹ , R ^2, and R ³ is an unsubstituted or substituted alkyl group, that is, an unsubstituted alkyl group having 1 to 2 carbon atoms (methyl group, ethyl group), or an alkyl moiety having a carbon number of A substituted alkyl group having 1 to 2 and substituted with an alkoxy group having 1 to 2 carbon atoms (methoxy group, ethoxy group), an alkyl moiety having 1 to 2 carbon atoms, and a phenyl group or p-methoxyphenyl A substituted alkyl group substituted with a group.

In the present specification, the “carbon number” of a substituent refers to the number of carbon atoms when the substituent is unsubstituted. Therefore, for example, when the group represented by R ¹ is a substituted alkyl group, the carbon number is the number of carbon atoms in the skeleton part of the alkyl group that does not include the carbon atom of the substituent that substitutes for the alkyl group. Say. The same applies to the case where the substituent is other than an alkyl group.

Specific examples of the alkyl group represented by R ¹ , R ² and R ³ include a benzyl group, a p-methoxybenzyl group, a methoxymethyl group and the like.
In the general formula (A), the substituent of the substituted silyl group represented by R ¹ , R ² and R ³ is an alkyl group having 1 to 4 carbon atoms (for example, methyl group, ethyl group, isopropyl group, t-butyl). Group) or an aryl group having 6 carbon atoms (for example, a phenyl group).

The substituted silyl group represented by R ¹ , R ² and R ³ is preferably a trisubstituted silyl group, more preferably a t-butyldimethylsilyl group, a triethylsilyl group, a triisopropylsilyl group, or the like. .

In consideration of using the compound represented by the general formula (A) as an intermediate of the sulfopyranosylacylglycerol derivative, the group represented by R ¹ , R ² and R ³ is a benzyl group, It is preferable from the viewpoint of stability as a protecting group. R ¹ , R ² and R ³ are p-methoxybenzyl group, t-butyldimethylsilyl group or triethylsilyl group for synthesizing a sulfopyranosylacylglycerol derivative to which an unsaturated fatty acid is bonded. It is preferable from the viewpoint of reactivity when deprotecting in further reaction.

In the general formula (A), R ⁴ represents an alkylsulfonyl group or an arylsulfonyl group.
The alkyl part of the alkylsulfonyl group is an alkyl group having 1 to 2 carbon atoms (methyl group or ethyl group). Specifically, the alkylsulfonyl group is a methanesulfonyl group or an ethanesulfonyl group.

  The aryl moiety of the arylsulfonyl group is an unsubstituted or substituted aryl group, that is, a phenyl group, or a substituted phenyl group substituted with a p-methyl group or a p-methoxy group. Specifically, the arylsulfonyl group includes a p-toluenesulfonyl group (tosyl group), a p-methoxybenzenesulfonyl group, a benzenesulfonyl group, and the like. Of these arylsulfonyl groups, a tosyl group is preferred from the viewpoint of reaction stability.

  The pyranoside (1-O- (2-propenyl) -6-deoxy-6-carbonylthiopyranoside) represented by the general formula (B) will be described in detail.

により表されるピラノシドの糖骨格を構成するピラノースは、上述した一般式（Ａ）により表されるピラノシドのそれと同義である。 Formula (B):

The pyranose constituting the sugar skeleton of the pyranoside represented by is synonymous with that of the pyranoside represented by the general formula (A).

In the general formula (B), the 2-propenyl group bonded to the C1 carbon may be an α bond or a β bond as in the case of the general formula (A).
In the general formula (B), R ¹ , R ² and R ³ are also synonymous with R ¹ , R ² and R ³ in the general formula (A).

In the general formula (B), R ⁵ represents a hydrogen atom, an alkyl group, or an aryl group.

The alkyl group represented by R ⁵ is an alkyl group having 1 to 2 carbon atoms (methyl group or ethyl group).

The aryl group represented by R ⁵ includes an aryl group having 6 carbon atoms (for example, a phenyl group).
In the compound represented by the general formula (B), the group represented by R ⁵ is preferably a methyl group from the viewpoint of reaction stability.

  By using the compounds represented by the general formula (A) and the general formula (B) as intermediates, the compound of the general formula (1) can be efficiently produced. That is, the compound of the general formula (1) can be produced through the following (Step A) to (Step J).

  (Step A) The hydroxyl group bonded to the C1 carbon of D-glucose that provides the sugar moiety of the compound represented by the general formula (1) is 2-propenylated. (Step B) Protect the hydroxyl group of C6 carbon of glucose. (Step C) Protect the hydroxyl group bonded to the C2, C3 and C4 carbons. (Step D) The C6 carbon protecting group previously protected is deprotected. (Step E) The hydroxyl group bonded to the C6 carbon is substituted with a group that can be converted into a carbonylthio group (for example, an alkylsulfonyloxy group or an arylsulfonyloxy group). (Step F) Carbonylation of C6 carbon. (Step G) The 2-propenyl group bonded to C1 is diolized. (Step H) At least one of the obtained diols is esterified with a desired linear higher fatty acid. (Step I) C6 carbon carbonylthio group is sulfonated. (Step J) By deprotecting the C2, C3 and C4 carbon protecting groups of the obtained sulfonate, the compound of the general formula (1) in the form of a salt can be produced. The salt thus obtained can be converted to a compound represented by the general formula (1) by subjecting it to titration with an acid such as hydrochloric acid.

  First, the manufacturing method (the said process AE) of the compound represented by general formula (A) and the manufacturing method (the said process F) of a compound represented by general formula (B) are demonstrated in detail.

  The pyranoside represented by the general formula (A) can be produced from the corresponding pyranose through the following five steps (steps A to E).

The pyranoside of the general formula (A) thus obtained can be converted into a pyranoside of the general formula (B) by further passing through the step F.

  In the method for producing the pyranoside of the general formula (A) through the above steps A to E, first, in step A, the hydroxyl group bonded to the C1 carbon of the corresponding unsubstituted pyranose (compound 1) is 2-propenylated, and the compound Get 2.

Next, in step B, the hydroxyl group of the C6 carbon of compound 2 is protected to obtain compound 3.
Next, in Step C, the hydroxyl group bonded to the C2, C3, and C4 carbons of Compound 3 is protected, and Compound 4 is obtained.

Subsequently, the protecting group couple | bonded with C6 carbon of the compound 4 is deprotected by the process D, and the compound 5 is obtained.
Finally, in Step E, a compound (compound 6) of the general formula (A) is obtained by bonding a leaving group for releasing an oxygen atom to the C6 carbon of compound 5.

  The pyranoside of the general formula (A) thus obtained can be further converted into a pyranoside of the general formula (B) through Step F.

The steps A to F will be described in more detail.
The 2-propenylation in step A is performed by reacting the corresponding pyranose and allyl alcohol in the presence of a strong acid such as trifluoromethanesulfonic acid, usually at room temperature to 100 ° C, preferably 80 ° C to 90 ° C. be able to.

In Step B, the hydroxyl group bonded to the C6 carbon is protected, and —OR ⁶ is bonded to the C6 carbon (wherein R ⁶ represents an alkyl group or a substituted silyl group).

The alkyl group represented by R ⁶ is a bulky unsubstituted or substituted alkyl group, and preferably includes an alkyl group having 1 to 4 carbon atoms (for example, a t-butyl group or a trityl group). The alkyl group represented by R ⁶ is preferably a trityl group from the viewpoint of ease of reaction.

The substituent of the substituted silyl group represented by R ⁶ includes a lower alkyl group, preferably an alkyl group having 1 to 4 carbon atoms (for example, methyl group, ethyl group, isopropyl group, t-butyl group), and aryl group Preferably an aryl group having 6 carbon atoms (for example, a phenyl group). The substituted silyl group represented by R ⁶ is preferably a trisubstituted silyl group, more preferably a t-butyldiphenylsilyl group.

  In step B, the hydroxyl group is protected by adding a compound capable of protecting the hydroxyl group such as trityl chloride to a solution of compound 2 dissolved in an organic solvent such as dry pyridine in the presence of a catalyst such as dimethylaminopyridine (DMAP). The reaction can be carried out at room temperature.

When trityl chloride is used as the compound capable of protecting the hydroxyl group, compound 3 in which R ⁶ is a trityl group is obtained. Trityl chloride can be preferably used from the viewpoint of production cost. Further, t-butyldiphenylsilyl chloride can be used as a compound capable of protecting a hydroxyl group, and can be reacted at room temperature in the presence of a catalyst such as imidazole. In this case, compound 3 in which R ⁶ is a t-butyldiphenylsilyl group is obtained.

In Step C, the hydroxyl groups bonded to the C2, C3 and C4 carbons are protected, and —OR ¹ , —OR ² and —OR ³ respectively (wherein R ^{1 to} R ³ are the same as those described above for the general formula (A)). Are synonymous with each other). These hydroxyl groups are protected by activating the hydroxyl group bonded to the C2, C3 and C4 carbons of compound 3 dissolved in an organic solvent such as dimethylformamide (DMF) with sodium hydride and the like to protect hydroxyl groups such as benzyl bromide. This can be done by reacting the resulting compound at room temperature.

When benzyl bromide is used as the compound capable of protecting the hydroxyl group, compound 4 in which R ¹ , R ² and R ³ are all benzyl groups is obtained. Benzyl bromide can be preferably used from the viewpoint of the stability of the protecting group. Moreover, p-methoxybenzyl bromide, t-butyldimethylsilyl chloride, triethylsilyl chloride and the like can also be used as a compound capable of protecting a hydroxyl group, and all of R ¹ , R ² and R ³ are each a p-methoxybenzyl group. , T-butyldimethylsilyl group, triethylsilyl group 4 is obtained. The reaction in the case of using a compound capable of protecting these hydroxyl groups can be carried out under reaction conditions suitable for each protecting group.

  The deprotection of the protecting group bonded to C6 carbon in Step D can be performed by reacting a solution of Compound 4 dissolved in an organic solvent such as methanol at room temperature in the presence of a catalyst such as toluenesulfonic acid.

In Step E, a leaving group (—OR ⁴ (same as defined in General Formula (A)) for releasing an oxygen atom is bonded to C6 carbon of Compound 5.
The leaving group (—OR ⁴ ) can be introduced into the C6 carbon by adding a compound capable of leaving an oxygen atom to a solution of the compound 5 dissolved in an organic solvent and reacting the compound.

In the above reaction, pyridine, dichloromethane or the like can be used as the organic solvent.
The above reaction can be performed at room temperature in the presence of a catalyst such as DMAP, if necessary.

As the compound capable of releasing an oxygen atom, a compound having an alkylsulfonyl group, a compound having an arylsulfonyl group, and the like can be used. The alkyl group of the compound having an alkylsulfonyl group is preferably an unsubstituted alkyl group, more preferably a lower alkyl group, still more preferably an alkyl group having 1 to 2 carbon atoms (methyl group, ethyl group). included. Specific examples of the compound having an alkylsulfonyl group include methanesulfonyl chloride, ethanesulfonyl chloride and the like. When methanesulfonyl chloride and ethanesulfonyl chloride are used, compound 6 in which the groups represented by R ⁴ are a methanesulfonyl group and an ethanesulfonyl group, respectively, is obtained.

The aryl group of the compound having an arylsulfonyl group used as a compound capable of leaving an oxygen atom is an unsubstituted or substituted aryl group, and preferably includes 6 carbon atoms (for example, a phenyl group). When the aryl group is substituted, the substituent includes a p-methyl group, a p-methoxy group, and the like. Specific examples of the compound having an arylsulfonyl group include p-toluenesulfonyl chloride, p-methoxybenzenesulfonyl chloride, benzenesulfonyl chloride and the like. When p-toluenesulfonyl chloride is used, compound 6 in which the group represented by R ⁴ is a p-toluenesulfonyl group (tosyl group) is obtained. When p-methoxybenzenesulfonyl chloride is used, compound 6 in which the group represented by R ⁴ is a p-methoxybenzenesulfonyl group is obtained.

  Of these compounds having an alkylsulfonyl group or arylsulfonyl group, those having a tosyl group are preferred from the viewpoint of easy reaction.

From the thus obtained pyranoside of the general formula (A) (compound 6), as shown in Step F, the sulfonyloxy group (—OR ⁴ ) of the compound 6 is converted to a carbonylthio group (—SC (═O)). By substituting R ⁵ ), the pyranoside of the general formula (B) (compound 7) can be produced.

  That is, in step F, a compound in which an alkyl or arylsulfonyloxy group can be substituted with a carbonylthio group on the pyranoside of the general formula (A) in an organic solvent (hereinafter referred to as “O-substituent → S-substituent compound”). The pyranoside of the general formula (B) can be produced by reacting.

  The O-substituent → S-substituent compound includes alkali metal salts and alkaline earth metal salts of thiocarboxylic acid. Examples of thiocarboxylic acids include thioformic acid and lower thiocarboxylic acids, preferably thiocarboxylic acids substituted with aliphatic groups having 1 to 2 carbon atoms (eg, thioacetic acid, thiopropionic acid), and thiocarboxylic acids substituted with aromatic groups having 6 carbon atoms. Acid (for example, thiobenzoic acid) and the like are included.

  Alkali metals that form salts with these thiocarboxylic acids include potassium, sodium, and the like, and alkaline earth metals include magnesium, calcium, and the like.

  Among the O-substituent → S-substituent compounds, the salt of thioacetic acid is used in terms of reaction stability, and the compound of the general formula (B) is used as an intermediate of the sulfopyranosylacylglycerol derivative. In view of the above, it can be preferably used from the viewpoint of easily leaving the carbonyl group in the subsequent step for producing the sulfopyranosyl acylglycerol derivative.

The addition amount of the O-substituent → S-substituent compound varies depending on the compound used, but can usually be set to an equivalent amount to twice the amount of the compound of the general formula (A).
Organic solvents used in the reaction include alcohols, preferably lower alcohols (eg, methanol, ethanol, propanol) and the like.

The amount of the organic solvent used can be usually set to about 2 to 10 times the amount from which the compound of the general formula (A) to be dissolved can be dissolved.
The above reaction can be usually carried out by stirring for 1 to 24 hours at room temperature to the boiling point of the solvent used.

  In addition, when the unsubstituted pyranose which is a starting material is a mixture of α-anomer and β-anomer, compound 6 and compound 7 are also a mixture of α- and β-anomer. These mixtures can be separated by converting to benzylidene derivatives or the like after Step A and crystallizing them, or subjecting them to chromatography after any of Steps A to F, if necessary.

By using the method in which the pyranoside represented by the general formula (B) thus produced is further subjected to the reaction in the next four steps (steps G to J), the general formula (1) in the form of a salt is used. Can be produced.

  In the above steps G to J, first, in step G, the allyl group of compound 7 (compound represented by formula (B)) is converted into a diol to obtain compound 8.

Next, in Step H, at least one of the diols of Compound 8 is esterified to obtain Compound 9. In compound 9, R ¹¹ represents a hydrogen atom or an acyl group. R ¹² represents an acyl group.

  Next, in Step I, the carbonylthio group of Compound 9 is sulfonated to obtain Compound 10.

  Finally, in Step J, the protecting group bonded to the C2-C4 carbon of Compound 10 is deprotected to obtain the target salt of the sulfopyranosylacylglycerol derivative (Compound 11).

The steps G to J will be described in detail.
Diolization in Step G is performed by adding an oxidizing agent such as osmium tetroxide to a solution of Compound 7 (compound represented by the general formula (B)) dissolved in a solvent mixture such as t-butanol and water, and trimethylamine N -It can be carried out by allowing a reoxidant such as oxide to coexist and reacting at room temperature.

  By the esterification reaction in Step H, a sulfopyranosyl acylglycerol derivative in which a desired fatty acid is ester-bonded with glycerol can be obtained. In this reaction, a fatty acid corresponding to the final product is added to a solution of Compound 8 dissolved in an appropriate organic solvent such as dichloromethane, and an appropriate solution such as ethyldimethylaminopropylcarbodiimide (EDCI) -DMAP system is added as necessary. It can carry out by making it react in presence of a catalyst.

According to the reaction in Step H, in compound 9, R ¹¹ is a hydrogen atom and R ¹² is a fatty acid acyl residue added with a monoester, and R ¹¹ and R ¹² are both acylated fatty acid residues added. A mixture of certain diesters is obtained.

As the fatty acid to be added, linear or branched, saturated or unsaturated fatty acid can be used. As the saturated fatty acid, a fatty acid having an acyl group represented by R ₁₀₁ in the general formula (1) described above can be used. One or more fatty acids can be added. When two or more fatty acids are added, R ¹¹ is a hydrogen atom, and monoester which is an acyl residue of any one of the fatty acids to which R ¹² is added, and any of the fatty acids to which R ¹¹ and R ¹² are added together A mixture with a diester which is one acyl residue is obtained.

  The mixture of these monoesters and diesters can be isolated to each ester by chromatography etc. as needed, and can use for reaction of the following process I.

For the monoester, R ¹¹ and R ¹² are different acyl residues by reacting a fatty acid having an acyl residue other than the acyl residue of R ¹² obtained in the above step H, if desired. Diesters can also be obtained. The reaction conditions for this further esterification can be set to the same conditions as those for step H except that the fatty acids are different.

Step I sulphonation involves the addition of an oxidant such as OXONE (2KHSO ₅ , KHSO ₄ , K ₂ SO ₄ ), etc. to a solution of compound 9 in an organic solvent buffered with glacial acetic acid and potassium acetate, The reaction can be carried out at room temperature.

  The deprotection of the protecting group bonded to the C2-C4 carbon in Step J is performed by removing a solution of Compound 10 dissolved in an organic solvent such as ethanol in a hydrogen gas atmosphere in the presence of a catalyst such as palladium-activated carbon (Pd-C). Can be performed at room temperature.

In general formula (A), in particular, a compound in which the group represented by R ^{1 to} R ³ is a substituted silyl group and the group represented by R ⁴ is an alkylsulfonyl group or an arylsulfonyl group includes the following three steps: It can also be manufactured through (Steps K to M).

The steps K to M will be described in detail.
Step K is the same as step A described above.
In Step L, a leaving group (R ⁴ ) for releasing an oxygen atom is bonded to the C6 carbon of compound 13. R ⁴ represents an alkylsulfonyl group or an arylsulfonyl group. R ⁴ is preferably an arylsulfonyl group.

The process L can be performed on the same conditions as the process E mentioned above.
In step M, a substituted silyl group is introduced into the C2-C4 carbon of compound 14. The substituted silyl group is preferably a trisubstituted silyl group, and more preferably t-butyldimethylsilyl, trimethylsilyl, triethylsilyl, triisopropylsilyl and the like. From the viewpoint of the stability of the product (Compound 15), t-butyldimethylsilyl is preferred.

  In this reaction, a compound capable of protecting a hydroxyl group such as t-butyldimethylsilyl trifluoromethanesulfonate is added to a solution of compound 14 dissolved in an organic solvent such as dry dichloromethane, and in the presence of a catalyst such as 2,6-lutidine. At room temperature.

  Thus, the compound represented by the corresponding general formula (B) can be produced from the compound represented by the general formula (A) obtained in the step M through the step F described above.

  In addition, a salt of the corresponding sulfopyranosylacylglycerol derivative can be produced from the compound represented by the general formula (B) obtained in the step F in this manner through the above-described steps G to J. .

  Furthermore, a sulfopyranosyl acylglycerol derivative can be produced by subjecting the sulfonate obtained in Step J to titration with an acid such as hydrochloric acid.

  Of the above-described sulfopyranosyl acylglycerol derivatives, the β-anomer of the sulfoquinovosyl acylglycerol derivative (the compound of the present invention represented by the general formula (2)) is a novel compound.

  This sulfoquinovosyl acylglycerol β derivative can be produced in the same manner as the compound of the general formula (1). However, in the above-mentioned method for producing the compound of the general formula (1), a step of separating a β-form from a mixture of α-form and β-form produced by using a mixture of α- and β-anomers of D-glucose as a starting material. Add The separation step can be performed at an appropriate time in the method for producing the compound of the general formula (1). For example, the separation step can be performed after the step D or the step F. The separation method can be carried out by silica gel chromatography known per se, for example, using a suitable solvent.

  Further, the sulfoquinovosyl acylglycerol β derivative of the present invention represented by the general formula (2) can also be produced by the following method. That is, the hydroxyl group bonded to all carbons of the sugar is acetylated, and then the C1 carbon is halogenated. By reacting this sugar halide with allyl alcohol, allyl alcohol is selectively β-bonded. 1-O- (2-propenyl) -β-D-glucose is obtained by deacetylating the obtained β-form product. These series of reactions are known reactions. The β derivative represented by the general formula (2) of the present invention uses 1-O- (2-propenyl) -β-D-glucose, which is a product of the above reaction, for the above-described Step B to Step J. Can also be manufactured.

  Hereinafter, the present invention will be described by way of examples. However, the present invention is not limited to these examples.

Physiological assay for the compound represented by the general formula (1) <Assay 1>
The inhibitory effect assay for DNA synthase α type was performed by the following method.

DNA synthase α-type 0.05 U purified from bovine thymus by antibody column and test compound (compound SQAG1, SQAG2, SQAG4, SQAG6, SQAG8, SQAG11, SQAG12, SQAG13 dissolved in DMSO as shown in Table 1 above) And SQAG14) were mixed, and an inorganic salt buffer solution necessary for enzyme reaction, [ ³ H] -labeled dTTP, and a reaction compound containing a template DNA strand were added, and incubated at 37 ° C. for 60 minutes.

After stopping the enzyme reaction, the product after the reaction was fixed on a dedicated filter and measured with a liquid scintillation counter. The amount of enzyme-synthesized dTTP was calculated as [ ³ H] radiation dose (cpm).

The obtained results are shown in Table 2 as IC ₅₀ .

  As is apparent from Table 2 above, all tested compounds have significant inhibitory activity against DNA synthase α-form.

  Colorectal cancer and gastric cancer cells used in the next two assays are examples of cancer cells in which the present anticancer drug can be effective. That is, these assays are not intended to limit the cancer cells in which the anticancer drug can be effective.

<Assay 2>
The anticancer test for colon cancer cultured cells was performed by the following method.

Colon cancer cell DLD-1 was maintained and passaged in RPMI 1640 medium (containing 10% calf serum). Test compounds (compounds SQAG1, SQAG2, SQAG4, SQAG6, SQAG8, SQAG11, SQAG12, SQAG13 and SQAG14 shown in Table 1 above) were suspended and diluted in the medium, respectively, and 96 wells with 3 × 10 ³ cells / well. Cultured in a petri dish. After culturing for 48 hours, MTT assay (Mosmann, T: Journal of immunological method, 65, 55-63 (1983)) was performed, and the survival rate was compared.

The obtained results are shown in Table 3 as IC ₅₀ .

  As is apparent from Table 3 above, all of the tested compounds have significant anticancer activity against colon cancer cells.

  Each of the compounds tested alone has the same or higher anticancer activity as the mixture of the sulfoquinovosyl acylglycerol derivatives disclosed in Sahara et al. It seems to have.

<Assay 3>
An anticancer test for gastric cancer cultured cells was performed in the same manner as assay 1 except that gastric cancer cell NUGC-3 was used instead of colon cancer cell DLD-1.

The obtained results are shown in the following Table 4 as IC ₅₀ .

  As is apparent from Table 4 above, all of the tested compounds have significant anticancer activity against gastric cancer cells.

  Each of the compounds tested alone has the same or higher anticancer activity as the sulfoquinovosylacylglycerol derivative mixture disclosed in Sahara et al. It seems that.

Synthesis Examples Production examples of compounds represented by general formula (A), general formula (B), general formula (1) and general formula (2) are shown below.

The following reaction scheme 1 is an example of a method for producing a compound represented by general formula (A), general formula (B), and general formula (1).

  In the above scheme 1, the synthesis route for only the α-anomer separated by silica gel flash chromatography after step d is shown, but for the β-anomer, a sulfopyranosylacylglycerol derivative is synthesized by the same reaction. Can do. Also, the mixture of monoester and diester obtained after step h can be separated by chromatography, and each ester can be subjected to step i.

The following scheme 2 is a reaction scheme suitable for the synthesis of the compounds of the general formulas (A) and (B) in which R ^{1 to} R ³ are substituted silyl groups and the corresponding sulfopyranosyl acylglycerol derivatives. In the reaction of Scheme 2, by passing through steps b ′ and c ′, only the α-anomer can be selectively synthesized without passing through the separation step performed after step d of reaction scheme 1.

<Example 1: Synthesis of compound represented by formula (A) (1)>
2,3,4-Tri-O-benzyl-1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose (VI) was synthesized from D-glucose.

1-1) Step a; Synthesis of 1-O- (2-propenyl) -D-glucose (II)

  100 g of D-glucose (I) was added to 250 mL of allyl alcohol and sufficiently dissolved, and 0.8 mL of trifluoromethanesulfonic acid was gradually added to the solution under ice cooling. Then, it was made to react for 30 hours, stirring at 80 degreeC in an oil bath. When the reaction was sufficiently advanced, the reaction mixture was neutralized with 1 mL of triethylamine and concentrated under reduced pressure. A production rate of about 60 to 70% was confirmed by thin layer chromatography.

1-2) Step b; Synthesis of 1-O- (2-propenyl) -6-O-triphenylmethyl-D-glucose (III)

  100 g (455 mmol) of 1-O- (2-propenyl) -D-glucose (II) is dissolved in 350 mL of dry pyridine, and 170 g (610 mmol) of trityl chloride and 1.0 g (8.20 mmol) of DMAP are added to the solution. The reaction was allowed to proceed for 36 hours at room temperature with stirring. Thereafter, the reaction was stopped by adding 800 mL of cold distilled water, extracted with ethyl acetate (500 mL × 3 times), the organic layers were combined, neutralized to pH 4 with dilute hydrochloric acid, and washed with saturated brine (500 mL × 2 times). Then, it was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel flash chromatography (dichloromethane: methanol = 20: 1). A production rate of about 80% was confirmed by thin layer chromatography.

1-3) Step c; Synthesis of 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-O-triphenylmethyl-D-glucose (IV)

  Take 80 g of sodium hydride (80 g, 83.3 mmol) diffused in mineral oil in a reactor, wash well with 50 mL of dry hexane, remove hexane, and dissolve 1-O- (2 -Propenyl) -6-O-triphenylmethyl-D-glucose (III) 10.0 g (21.6 mmol) was gradually added under ice-cooling. After 15 minutes, the mixture was returned to room temperature and reacted for 1 hour with stirring. I let you.

  Next, 12.0 g (70.2 mmol) of benzyl bromide was gradually added again under ice-cooling, and after 15 minutes, the temperature was returned to room temperature and reacted for 3 hours with stirring. Then, 20 mL of methanol and 30 mL of cold distilled water were added to stop the reaction, and the mixture was extracted with ethyl acetate (50 mL × 3 times). The organic layers were combined and washed with saturated brine (100 mL × 2 times), and then anhydrous sodium sulfate. And filtered, concentrated under reduced pressure, and purified by silica gel flash chromatography (hexane: ethyl acetate = 10: 1) (yield 9.6 g (13.8 mmol), yield 63.9%).

1-4) Step d; Synthesis of 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -α-D-glucose (V)

  9.6 g (13.8 mmol) of 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-O-triphenylmethyl-D-glucose (IV) was dissolved in 100 mL of methanol. , 3.8 g (20.0 mmol) of p-toluenesulfonic acid monohydrate was added and allowed to react for 16 hours with stirring. Thereafter, the reaction was stopped by adding 100 mL of cold distilled water, extracted with ethyl acetate (200 mL × 3 times), the organic layers were combined, washed with saturated brine (300 mL × 2 times), and then dried over anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and separation and purification of α form and β form by silica gel flash chromatography (hexane: ethyl acetate = 11: 2 → 4: 1 → 2: 1) (yield 2.70 g (5.50 mmol) of α form) ), Yield 39.8%, β-form yield 1.52 g (3.10 mmol), yield 22.5%).

1-5) Step e; Synthesis of 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose (VI)

10.0 g (20.4 mmol) of 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -α-D-glucose (V) was dissolved in 200 mL of dry pyridine, and 134 mg (1. 10 mmol) and 9.2 g (48.3 mmol) of p-toluenesulfonyl chloride were added and reacted at room temperature for 16 hours with stirring. Then, the reaction was stopped by adding 300 mL of cold distilled water, extracted with ethyl acetate (200 mL × 3 times), the organic layers were combined, neutralized to pH 4 with dilute hydrochloric acid, and washed with saturated brine (300 mL × 2 times). Then, it was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel flash chromatography (hexane: ethyl acetate = 4: 1) (yield 12.0 g (18.6 mmol), yield 91.2%). Melting point 77-79 ° C., [α] _D = + 51.8 ° (CHCl ₃ ).

1 and 2 show NMR charts.
FIG. 1 is a chart of ¹ H NMR (300 MHz, CDCl ₃ ). Tetramethylsilane was used as an internal standard substance.
FIG. 2 is a chart of ¹³ C NMR (300 MHz, CDCl ₃ ).

<Example 2: Synthesis of compound represented by general formula (A) (2)>
From D-glucose (I ′), 2,3,4-tri-O- (t-butyldimethylsilyl) -1-O (2-propenyl) -6-O— is obtained by the following steps a ′ to e ′. (4-Tolylsulfonyl) -α-D-glucose (VI ′) was synthesized.

2-1) Step a ′; Synthesis of 1-O- (2-propenyl) -D-glucose (II ′)

  100 g of D-glucose (I ′) was added to 250 mL of allyl alcohol and sufficiently dissolved, and 0.8 mL of trifluoromethanesulfonic acid was gradually added to the solution under ice cooling. Then, it was made to react for 30 hours, stirring at 80 degreeC in an oil bath. When the reaction was sufficiently advanced, the reaction mixture was neutralized with 1 mL of triethylamine and concentrated under reduced pressure. A production rate of about 60 to 70% was confirmed by thin layer chromatography.

2-2) Step b ′; Synthesis of 1-O- (2-propenyl) -4,6-O-benzylidene-α-D-glucose (III ′)

  37.5 g of 1-O- (2-propenyl) -D-glucose (II ′) was sufficiently dissolved in 210 mL of benzaldehyde, and 98 g of zinc chloride was added to the solution and reacted at room temperature for 4 hours. Thereafter, the reaction solution was added to 500 mL of hexane, and further 100 mL of dilute sodium hydrogen carbonate solution was added, and the mixture was allowed to stand at 0 ° C. for 30 minutes for crystallization. The crystals were subjected to suction filtration, dissolved in 50 mL of ethanol, and allowed to stand at 0 ° C. for 30 minutes for recrystallization (yield 21 g (68.1 mmol), yield 40.0%).

2-3) Step c ′; Synthesis of 1-O- (2-propenyl) -α-D-glucose (IV ′)

  10.7 g (34.7 mmol) of 1-O- (2-propenyl) -4,6-O-benzylidene-α-D-glucose (III ′) was dissolved in 260 mL of a solution of acetic acid: water = 8: 5, After reacting at 100 ° C. for 1 hour, the mixture was concentrated under reduced pressure and purified by silica gel flash chromatography (dichloromethane: methanol = 6: 1) (yield 6.3 g (28.6 mmol), yield 82.4%).

2-4) Step d ′; Synthesis of 1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose (V ′)

  6.3 g (28.6 mmol) of 1-O- (2-propenyl) -α-D-glucose (IV ′) was dissolved in 200 mL of dry pyridine, 195 mg of DMAP and 7.0 g of p-toluenesulfonyl chloride were added and stirred. The reaction was allowed to proceed for 16 hours at room temperature. Thereafter, the reaction was stopped by adding 20 mL of cold distilled water, extracted with ethyl acetate (200 mL × 3 times), the organic layers were combined, neutralized to pH 4 with 1.0 N and 0.1 N hydrochloric acid, and saturated with brine. After washing (200 mL × 2 times), drying over anhydrous sodium sulfate, filtration, concentration under reduced pressure, and purification by silica gel flash chromatography (dichloromethane: methanol = 20: 1) (yield 8.6 g (24.0 mmol), yield 83.8%).

2-5) Step e ′; 2,3,4-tri-O- (t-butyldimethylsilyl) -1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D -Synthesis of glucose (VI ')

11.2 g (29.9 mmol) of 1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose (V ′) was dissolved in 25 mL of dry dichloromethane, and t-butyldimethyl was dissolved. 23.8 g of silyl trifluoromethanesulfonate and 14.4 g of 2,6-lutidine were added and reacted for 16 hours with stirring under a nitrogen stream. Then, 150 mL of dichloromethane was added to stop the reaction, washed with saturated brine (100 mL × 2 times), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and silica gel flash chromatography (hexane: ethyl acetate = 30: 1). ) To give a colorless and transparent oily substance (yield 19.6 g (27.4 mmol), yield 91.6%). _{[Α] D = + 39.0 °} (CHCl 3).

3 and 4 show NMR charts.
FIG. 3 is a chart of ¹ H NMR (300 MHz, CDCl ₃ ). Tetramethylsilane was used as an internal standard substance.
FIG. 4 is a chart of ¹³ C NMR (300 MHz, CDCl ₃ ).

<Example 3: Synthesis of compound represented by general formula (B) (1)>
From 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose (VI) obtained in Example 1 above, 2,3,4-Tri-O-benzyl-1-O- (2-propenyl) -6-deoxy-6-thioacetyl-α-D-glucose (VII) was synthesized by step f.

11.4 g (18.6 mmol) of 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose (VI) It melt | dissolved in 250 mL of dry ethanol, 5.6 g (49.0 mmol) of potassium thioacetate was added, and it was made to react for 3 hours, stirring under recirculation | reflux conditions. Thereafter, the reaction was stopped by adding 300 mL of cold distilled water, extracted with ethyl acetate (200 mL × 3 times), the organic layers were combined and washed with saturated brine (300 mL × 2 times), then dried over anhydrous sodium sulfate, Filtration, concentration under reduced pressure, and purification by silica gel flash chromatography (hexane: ethyl acetate = 10: 1) (yield 9.00 g (16.4 mmol), yield 88.2%). Melting point: 61-62.5 ° C., [α] _D = + 51.8 ° (CHCl ₃ ).

5 and 6 show NMR charts.
FIG. 5 is a chart of ¹ H NMR (300 MHz, CDCl ₃ ). Tetramethylsilane was used as an internal standard substance.
FIG. 6 is a chart of ¹³ C NMR (300 MHz, CDCl ₃ ).

<Example 4: Synthesis of compound represented by formula (B) (2)>
2,3,4-Tri-O- (t-butyldimethylsilyl) -1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D- obtained in Example 2 above From glucose (VI ′), according to step f ′, 2,3,4-tri-O- (t-butyldimethylsilyl) -1-O- (2-propenyl) -6-deoxy-6-thioacetyl-α- D-glucose (VII ′) was synthesized.

2,3,4-tri-O- (t-butyldimethylsilyl) -1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose (VI ′) 9 g (11.0 mmol) was dissolved in 20 mL of dry ethanol, 1.8 g of potassium thioacetate was added, and the mixture was reacted for 3 hours with stirring under reflux conditions. Thereafter, the reaction was stopped by adding 100 mL of cold distilled water, extracted with ethyl acetate (200 mL × 3 times), the organic layers were combined and washed with saturated brine (200 mL × 2 times), and then dried over anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification by silica gel flash chromatography (hexane: ethyl acetate = 50: 1) gave a colorless transparent oily substance (yield 5.6 g (9.02 mmol), yield 82.0%). , [Α] _D = + 60.9 ° (CHCl ₃ ).

7 and 8 show NMR charts.
FIG. 7 is a chart of ¹ H NMR (300 MHz, CDCl ₃ ). Tetramethylsilane was used as an internal standard substance.
FIG. 8 is a chart of ¹³ C NMR (300 MHz, CDCl ₃ ).

<Example 5: Synthesis of sulfoquinovosyl acylglycerol derivative (1)>
From 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-deoxy-6-thioacetyl-α-D-glucose (VII) obtained in Example 3 above, steps g to A sulfoquinovosyl acylglycerol derivative was synthesized by j.

5-1) Step g; Synthesis of 3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-thioacetyl-α-D-glucopyranosyl) -glycerol (VIII)

2,3,4-Tri-O-benzyl-6-deoxy-1-O- (2-propenyl) -6-thioacetyl-α-D-glucose (VII) 8.30 g (15.1 mmol) was added to t-butanol. : H ₂ O = 4: 1 Dissolved in a solution, 2.5 g (22.5 mmol) of trimethylamine N-oxide dihydrate and 20 mL of osmium tetroxide-t-butanol solution (0.04 M) were added and stirred. The reaction was allowed to proceed for 30 hours at room temperature. Thereafter, 15 g of activated carbon was added, and the mixture was allowed to stand at room temperature for 1.5 hours with stirring to adsorb osmium tetroxide, followed by suction filtration. Next, 250 mL of cold distilled water was added to stop the reaction, extracted with ethyl acetate (200 mL × 3 times), the organic layers were combined, washed with saturated brine (300 mL × 2 times), and then dried over anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification by silica gel flash chromatography (hexane: ethyl acetate = 1: 1) (yield 5.00 g (8.59 mmol), yield 56.9%).

5-2) Step h; 3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-thioacetyl-α-D-glucopyranosyl) -1,2-di-O-palmitoylglycerol ( IX-1) and 3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-thioacetyl-α-D-glucopyranosyl) -1-O-palmitoylglycerol (IX-2)

Here, IX-1: R ₁₁ = R ₁₂ = palmitate; IX-2: R ₁₁ = H, R ₁₂ = palmitate.

  20.3 mg (34.3 μmol) of 3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-thioacetyl-α-D-glucopyranosyl) -glycerol (VIII) was dissolved in 5 mL of dichloromethane. EDCI 19.4 mg (101 μmol), DMAP 5.70 mg (46.7 μmol) and 14.1 mg (54.9 μmol) palmitic acid were added, and the mixture was reacted at room temperature for 16 hours with stirring. Then, the reaction was stopped by adding 20 mL of dichloromethane, washed with saturated brine (20 mL × 2 times), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and silica gel flash chromatography (hexane: ethyl acetate = 7: 1 → 3: 1) to separate and purify the diester and monoester (yield diester 14.7 mg (13.9 μmol); monoester 9.10 mg (11.1 μmol), yield (both combined) 72.9%).

5-3-1) Step i-1; 3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-sulfo-α-D-glucopyranosyl) -1,2-di-O -Synthesis of palmitoylglycerol sodium salt (X-1)

Where R ₁₁ = R ₁₂ = palmitate.

  133 mg (125 μmol) of 3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-thioacetyl-α-D-glucopyranosyl) -1,2-di-O-palmitoylglycerol (IX-1) ) Was dissolved in 7 mL of glacial acetic acid, 814 mg of potassium acetate and 228 mg of OXONE were added, and the mixture was reacted at room temperature for 16 hours with stirring. Thereafter, the reaction was stopped by adding 20 mL of cold distilled water, extracted with ethyl acetate (20 mL × 5 times), the organic layers were combined and neutralized with saturated sodium bicarbonate solution (70 mL × 5 times), and then saturated brine (60 mL × 2 times), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel flash chromatography (dichloromethane: methanol = 10: 1) (yield 57.9 mg (13.9 μmol)). (Rate 43.4%).

5-3-2) Step i-2; 3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-sulfo-α-D-glucopyranosyl) -1-O-palmitoylglycerol Synthesis of sodium salt (X-2)

Here, R ₁₁ = H, R ₁₂ = palmitate.

  3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-thioacetyl-α-D-glucopyranosyl) -1-O-palmitoylglycerol (IX-2) 52.1 mg (63.5 μmol) ) Was dissolved in 2 mL of glacial acetic acid, 102 mg of potassium acetate and 116 mg of OXONE were added, and the mixture was reacted at room temperature for 16 hours with stirring. Thereafter, the reaction was stopped by adding 15 mL of cold distilled water, extracted with ethyl acetate (20 mL × 5 times), the organic layers were combined and neutralized with saturated sodium bicarbonate solution (70 mL × 5 times), and then saturated brine (60 mL × twice), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel flash chromatography (dichloromethane: methanol = 10: 1) (yield 35.1 mg (42.4 μmol)). Rate 66.8%).

5-4-1) Step j-1; 3-O- (6-deoxy-6-sulfo-α-D-glucopyranosyl) -1,2-di-O-palmitoylglycerol sodium salt (XI-1) Composition

Where R ₁₁ = R ₁₂ = palmitate.

3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-sulfo-α-D-glucopyranosyl) -1,2-di-O-palmitoylglycerol sodium salt (X-1) 359 mg (330 μmol) was dissolved in 50 mL of ethanol, 1.30 g of Pd-C was added, the inside of the flask was replaced with H ₂ , and the mixture was reacted at room temperature for 16 hours with stirring. Thereafter, the mixture was filtered with suction, concentrated under reduced pressure, and purified by silica gel flash chromatography (dichloromethane: methanol = 10: 1 → dichloromethane: methanol: water = 65: 25: 4) (yield 129 mg (168 μmol), yield 50.9). %).

5-4-2) Step j-2; Synthesis of 3-O- (6-deoxy-6-sulfo-α-D-glucopyranosyl) -1-O-palmitoylglycerol sodium salt (XI-2)

Here, R ₁₁ = H, R ₁₂ = palmitate.

3-O- (2,3,4-tri-O-benzyl-6-deoxy-6-sulfo-α-D-glucopyranosyl) -1-O-palmitoylglycerol sodium salt (X-2) 202 mg (238 μmol) Was dissolved in 25 mL of ethanol, 1.00 g of Pd-C was added, the inside of the flask was replaced with H ₂ , and the mixture was reacted at room temperature for 16 hours with stirring. Thereafter, the mixture was filtered with suction, concentrated under reduced pressure, and purified by silica gel flash chromatography (dichloromethane: methanol = 10: 1 → dichloromethane: methanol: water = 65: 25: 4) (yield 57.2 mg (168 μmol), yield 43 .3%).

<Example 6: Synthesis of sulfoquinovosyl acylglycerol derivative (2)>
2,3,4-Tri-O- (t-butyldimethylsilyl) -6-deoxy-1-O- (2-propenyl) -6-thioacetyl-α-D-glucose (VII) obtained in Example 4 above From '), a sulfoxynovosyl acylglycerol derivative was synthesized by steps g' to j '.

6-1) Step g ′; 3-O- [2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-6-thioacetyl-α-D-glucopyranosyl] -glycerol (VIII ′ ) Synthesis

  5.6 g (9 of 2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-1-O- (2-propenyl) -6-thioacetyl-α-D-glucose (VII ′) .02 mmol) is dissolved in a t-butanol: water = 4: 1 solution, 1.5 g of trimethylamine N-oxide dihydrate and 15 mL of an osmium tetroxide t-butanol solution (0.04M) are added and stirred. The reaction was allowed to proceed at room temperature for 22 hours. Thereafter, 15 g of activated carbon was added, and the mixture was allowed to stand at room temperature for 1.5 hours with stirring to adsorb osmium tetroxide, followed by suction filtration. Next, the reaction was stopped by adding 200 mL of cold distilled water, extracted with ethyl acetate (200 mL × 3 times), the organic layers were combined, washed with saturated brine (300 mL × 2 times), and then dried over anhydrous sodium sulfate. Filtration, concentration under reduced pressure, and purification by silica gel flash chromatography (hexane: ethyl acetate = 3: 1 → 2: 1) (yield 5.2 g (7.94 mmol), yield 88.0%).

6-2) Step h ′; 3-O- [2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-6-thioacetyl-α-D-glucopyranosyl] -1,2- Di-O-oleoyl-glycerol (IX′-1) and 3-O- [2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-6-thioacetyl-α-D- Synthesis of “glucopyranosyl” -1-O-oleoyl-glycerol (IX′-2)

Here, IX′-1: R ₁₁ = R ₁₂ = oleoate; IX′-2: R ₁₁ = H, R ₁₂ = oleoate.

  3-O- [2,3,4-Tri-O- (t-butyldimethylsilyl) -6-deoxy-6-thioacetyl-α-D-glucopyranosyl] -glycerol (VIII ′) 1.37 g (2.09 mmol) ) Was dissolved in 20 mL of dry dichloromethane, and 1.46 g of EDCI, 538 mg of DMAP, and 660 mg of oleic acid were added and reacted at room temperature for 16 hours with stirring. Then, 200 mL of dichloromethane was added to stop the reaction, washed with saturated brine (100 mL × 2 times), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and silica gel flash chromatography (hexane: ethyl acetate = 20: 1 → 10). 1 → 7: 1) (yield diester 772 mg (652 μmol); monoester 895 mg (974 μmol), yield (both combined) 78.0%).

6-3-1) Step i′-1; 3-O- [2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-6-sulfo-α-D-glucopyranosyl]- Synthesis of 1,2-di-O-oleoyl-glycerol sodium salt (X'-1)

Where R ₁₁ = R ₁₂ = oleoate.

  3-O- [2,3,4-Tri-O- (t-butyldimethylsilyl) -6-deoxy-6-thioacetyl-α-D-glucopyranosyl] -1,2-di-O-oleoyl-glycerol (IX′-1) 566 mg (478 μmol) was dissolved in glacial acetic acid 28 mL, potassium acetate 3.2 g and OXONE 980 mg were added, and the mixture was reacted at room temperature for 6 hours with stirring. The reaction was then stopped by adding 15 mL of cold distilled water, extracted with ethyl acetate (20 mL × 5 times), and the organic layers were combined and neutralized with saturated sodium bicarbonate solution (70 mL × 5 times), and then with saturated brine. After washing (2 × 60 mL), drying over anhydrous sodium sulfate, filtration, concentration under reduced pressure, and purification by silica gel flash chromatography (dichloromethane: methanol = 50: 1 → 10: 1) (yield 152 mg (126 μmol), yield 26.4%).

6-3-2) Step i′-2; 3-O- [2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-6-sulfo-α-D-glucopyranosyl]- Synthesis of 1-O-oleoyl-glycerol sodium salt (X'-2)

Here, R ₁₁ = H, R ₁₂ = oleoate.

  3-O- [2,3,4-Tri-O- (t-butyldimethylsilyl) -6-deoxy-6-thioacetyl-α-D-glucopyranosyl] -1-O-oleoyl-glycerol (IX′- 2) 21.4 mg (23.2 μmol) was dissolved in 3.5 mL of glacial acetic acid, 500 mg of potassium acetate and 35.4 mg of OXONE were added, and the mixture was reacted at room temperature for 6 hours with stirring. The reaction was then stopped by adding 15 mL of cold distilled water, extracted with ethyl acetate (20 mL × 5 times), and the organic layers were combined and neutralized with saturated sodium bicarbonate solution (70 mL × 5 times), and then with saturated brine. After washing (2 × 60 mL), drying over anhydrous sodium sulfate, filtration, concentration under reduced pressure, and purification by silica gel flash chromatography (dichloromethane: methanol = 50: 1 → 20: 1) (yield 7.70 mg (8.13 μmol) ), Yield 34.9%).

6-4-1) Step j′-1; 3-O- (6-deoxy-6-sulfo-α-D-glucopyranosyl) -1,2-di-O-oleoyl-glycerol sodium salt (XI ′ -1) Synthesis

Where R ₁₁ = R ₁₂ = oleoate.

  3-O- [2,3,4-Tri-O- (t-butyldimethylsilyl) -6-deoxy-6-sulfo-α-D-glucopyranosyl] -1,2-di-O-oleoyl-glycerol 214 mg (176 μmol) of sodium salt (X′-1) was dissolved in 5 mL of a solution of acetic acid: tetrahydrofuran: trifluoroacetic acid: water = 3: 1: 0.4: 1 and reacted at room temperature for 16 hours with stirring. Extracted with ethyl acetate (10 mL × 3), combined organic layers were washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and flash chromatography on silica gel (dichloromethane: methanol = 10: 1 → dichloromethane: methanol: water = 65: 25: 4) (yield 84.1 mg (99.1 μmol), yield 56.3%).

6-4-2) Step j′-2; 3-O- (6-deoxy-6-sulfo-α-D-glucopyranosyl) -1-O-oleoyl-glycerol sodium salt (XI′-2) Composition

Here, R ₁₁ = H, R ₁₂ = oleoate.

  3-O- [2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-6-sulfo-α-D-glucopyranosyl] -1-O-oleoyl-glycerol sodium salt ( X′-2) 358 mg (378 μmol) was dissolved in 7 mL of a solution of acetic acid: tetrahydrofuran: trifluoroacetic acid: water = 3: 1: 0.4: 1 and reacted at room temperature for 16 hours with stirring. Extracted with ethyl acetate (10 mL × 3), combined organic layers were washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and flash chromatography on silica gel (dichloromethane: methanol = 10: 1 → dichloromethane: methanol: water = 65: 25: 4) (yield 138 mg (237 μmol), yield 62.7%).

<Example 7: Synthesis of compound represented by general formula (2) of the present invention>
3-O- (6-deoxy-6-sulfo-β-D-glucopyranosyl) -1-O-palmitoylglycerol sodium salt was synthesized as follows.

  That is, 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -β-D-glucose separated after step d of Example 1 above. Was used to obtain the title compound as white crystals in the same manner as in Step f of Example 3 and Steps g, h, i-2 and j-2 of Example 5 (yield 1.52 g (3.10 mmol)). ), Yield 22.5%).

Melting point 80-82 ° C., [α] _D = + 0.4 ° (CHCl ₃ ).

  9 and 10 show NMR charts.

FIG. 9 is a chart of ¹ H NMR (300 MHz, CDCl ₃ ). Tetramethylsilane was used as an internal standard substance.

FIG. 10 is a chart of ¹³ C NMR (300 MHz, CDCl ₃ ).

¹ H NMR chart of 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose prepared in Example 1 . ¹³ C NMR chart of 2,3,4-tri-O-benzyl-1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose prepared in Example 1 . 2,3,4-Tri-O- (t-butyldimethylsilyl) -1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose prepared in Example 2 < ¹ > H NMR chart. 2,3,4-Tri-O- (t-butyldimethylsilyl) -1-O- (2-propenyl) -6-O- (4-tolylsulfonyl) -α-D-glucose prepared in Example 2 ¹³ C NMR chart. ¹ H NMR chart of 2,3,4-tri-O-benzyl-6-deoxy-1-O- (2-propenyl) -6-thioacetyl-α-D-glucose prepared in Example 3. FIG. ¹³ C NMR chart of 2,3,4-tri-O-benzyl-6-deoxy-1-O- (2-propenyl) -6-thioacetyl-α-D-glucose prepared in Example 3. FIG. Of 2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-1-O- (2-propenyl) -6- (thioacetyl) -α-D-glucose prepared in Example 4 ¹ H NMR chart. Of 2,3,4-tri-O- (t-butyldimethylsilyl) -6-deoxy-1-O- (2-propenyl) -6- (thioacetyl) -α-D-glucose prepared in Example 4 ¹³ C NMR chart. ¹ H NMR chart of 3-O- (6-deoxy-6-sulfo-β-D-glucopyranosyl) -1-O-palmitoylglycerol sodium salt prepared in Example 7. FIG. ¹³ C NMR chart of 3-O- (6-deoxy-6-sulfo-β-D-glucopyranosyl) -1-O-palmitoylglycerol sodium salt prepared in Example 7

Claims (2)

The following general formula (2):

(Wherein, R ₁₀₁ represents an acyl residue of a saturated higher fatty acid, R ₁₀₂ represents. An acyl residue of a hydrogen atom or a saturated higher fatty acid) Suruhokino having sulfonate group structures are in the form of a salt Bosyl acylglycerol. In the general formula (2), the acyl residues of the saturated higher fatty acids represented by R ₁₀₁ and R ₁₀₂ are each independently CH ₃ (CH ₂ ) _n CO— (n is an integer of 12 to 24). The sulfoquinovosyl acylglycerol according to claim 1 represented.

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