JP4749339B2 - Cellooligosaccharide derivative and method for producing the same - Google Patents

Description

  The present invention relates to a cellooligosaccharide derivative and a method for producing the same.

  In recent years, surfactants have been used for a very wide range of applications. Among them, ionic surfactants may have problems in terms of safety, stability, biodegradability, etc., depending on the intended use. On the other hand, the nonionic surfactant has an excellent feature that the above problems are reduced. For this reason, research on such nonionic surfactants is energetically performed today.

  Until now, many nonionic surfactants have been produced and used, and for example, sucrose fatty acid esters, sorbitan monostearate and the like are known. These nonionic surfactants contain components such as long-chain fatty acids in addition to the sugar moiety in the molecule, thereby maintaining a balance between hydrophilicity and hydrophobicity. However, there are few reports on surfactants composed of sugar chains that do not contain long-chain fatty acids.

  Slightly, Patent Document 1 describes that a cello-oligomer derivative obtained by partially methylating the hydroxyl group at the 6-position of cellobiose or cellooligosaccharide is useful as a surfactant. This cello-oligomer derivative is characterized in that the hydrophobic methyl group is uniformly substituted with a hydrophilic glucose ring, which is said to increase the surface activity.

  However, the cello-oligomer derivative of Patent Document 1 is synthesized using an enzyme (cellulase), and only cellooligosaccharides in which methyl groups are alternately introduced at the 6-position can be synthesized due to the substrate specificity of the enzyme used. There is a drawback. Further, Patent Document 1 does not describe the result of the evaluation of the surface activity of the cello-oligomer derivative, and it is not clear whether it actually has the surface activity.

In addition, there has been no report on cellooligosaccharides in which the introduction position of the hydrophobic group is biased along the cellulose main chain, and the solution properties are extremely interesting.
JP 7-138278 A

  It is an object of the present invention to provide a cellooligosaccharide derivative having a bias in the introduction position of a hydrophobic group along the sugar chain, a method for producing the same, and a nonionic surfactant and nanoparticles comprising the cellooligosaccharide derivative. .

  As a result of intensive studies to solve the above-mentioned problems, the present inventor has no long-chain hydrophobic group (C5 or higher), a monosaccharide having a hydrophobic part composed of methylated glucose or cellooligosaccharide and a hydroxyl group Or it discovered that the cellooligosaccharide derivative containing the hydrophilic part which consists of oligosaccharides in a block form melt | dissolves in water, methanol, chloroform, etc., and shows amphiphilicity. It was also found that this cellooligosaccharide derivative has characteristics as a nonionic surfactant that forms micelles in water and reverse micelles in chloroform. As a result of further research based on this knowledge, the present invention has been completed.

  That is, the present invention provides the following cellooligosaccharide derivatives and methods for producing the same, as well as nonionic surfactants and nanoparticles comprising the cellooligosaccharide derivatives.

  Item 1. A cellooligosaccharide derivative in which a hydrophobic moiety comprising an alkylated glucose or cellooligosaccharide and a hydrophilic moiety comprising a monosaccharide or oligosaccharide are bound in a block manner.

Item 2. General formula:
H- (BA) _x -OR ¹
HA- (BA) _x -OR ^{1
_{R 1 - (A-B)}} x -OH or _{^{R 1 -B- (A-B)}} x -OH
(In the formula, A is a hydrophobic moiety obtained by removing the anomeric hydroxyl group from glucose in which the hydroxyl groups at positions 2, 3, 6 are C _1-4 alkylated and -H is removed from the alcoholic hydroxyl group at position 4. Alternatively, the anomeric hydroxyl group of the reducing terminal sugar residue is removed from the cellooligosaccharide in which the hydroxyl groups at the 2, 3, 6 positions of each glucose residue are C _1-4 alkylated and the 4th position of the non-reducing terminal sugar residue. Is a hydrophobic part obtained by removing -H from the alcoholic hydroxyl group, and B is a hydrophilic part obtained by removing the anomeric hydroxyl group from a monosaccharide and removing -H from one alcoholic hydroxyl group, or A hydrophilic moiety obtained by removing an anomeric hydroxyl group of a reducing terminal sugar residue from an oligosaccharide and removing -H from one alcoholic hydroxyl group of a non-reducing terminal sugar residue, and containing two or more A Case A is the same or different from each other May have I, when containing 2 or more B B may be the same or different, x is an integer of 1 to 20, R ¹ represents H or C _1-4 alkyl. However, sugar The chain is placed so that the reducing end is on the right side of the page.)
Item 2. A cellooligosaccharide derivative according to item 1.

Item 3. General formula:
^{H-B 1} -A ¹ -OR ¹ or R ¹ -A ¹ -B ¹ -OH
(In the formula, A ¹ is a hydrophobic moiety obtained by removing the anomeric hydroxyl group from glucose in which the hydroxyl groups at the 2, 3, and 6 positions are C _1-4 alkylated and removing -H from the alcoholic hydroxyl group at the 4 position. Alternatively, the anomeric hydroxyl group of the reducing terminal sugar residue is removed from the cellooligosaccharide in which the 2, 3, 6-position hydroxyl group of each glucose residue is C _1-4 alkylated, and 4 of the non-reducing terminal sugar residue. A hydrophobic moiety obtained by removing -H from the alcoholic hydroxyl group at the position, and B ¹ is a hydrophilic moiety obtained by removing the anomeric hydroxyl group from a monosaccharide and removing -H from one alcoholic hydroxyl group; Or a hydrophilic moiety obtained by removing the anomeric hydroxyl group of the reducing terminal sugar residue from the oligosaccharide and removing -H from one alcoholic hydroxyl group of the non-reducing terminal sugar residue, and R ¹ is H or an _{C 1-4} alkyl However, the sugar chain is arranged to be the reducing end on the right side.) Cellooligosaccharide derivative according to claim 1 or 2 represented by.

Item 4. General formula:
R ¹ -A ¹ -B ¹ -A ² -OR ^{1
R 1 -A 1 -B 1 -A 2} -B 2 -OH
R ¹ -A ¹ -B ¹ -A ² -B ³ -A ³ -OR ¹
R ¹ -A ¹ -B ¹ -A ² -B ² -A ³ -B ³ -OH
^{H-B 1 -A 1 -B 2} -OH
^{H-B 1 -A 1 -B 2} -A 2 -OR 1
^{H-B 1 -A 1 -B 2} -A 2 -B 3 -OH or ^{H-B 1 -A 1 -B 2} -A 2 -B 3 -A 3 -OR 1
(In the formula, A ¹ , A ² and A ³ are the same or different and are obtained by removing the anomeric hydroxyl group from glucose in which the hydroxyl groups at the 2, 3 and 6 positions are C _1-4 alkylated and the alcoholic hydroxyl group at the 4 position. The hydrophobic moiety obtained by removing -H from the above, or the anomeric hydroxyl group of the reducing terminal sugar residue from the cellooligosaccharide in which the hydroxyl groups at the 2, 3, 6 positions of each glucose residue are C _1-4 alkylated And a hydrophobic moiety obtained by removing -H from the alcoholic hydroxyl group at the 4-position of the non-reducing terminal sugar residue, and B ¹ , B ² and B ³ are the same or different and are anomalous from a monosaccharide Hydrophilic moiety obtained by removing hydroxyl group and removing -H from one alcoholic hydroxyl group, or one of non-reducing terminal sugar residues by removing anomeric hydroxyl group of reducing terminal sugar residue from oligosaccharide -H from the alcoholic hydroxyl group of And R ¹ represents H or C _1-4 alkyl, except that the sugar chain is arranged so that the reducing end is on the right side of the page.
Item 3. The cellooligosaccharide derivative according to item 1 or 2,

  Item 5. The hydrophobic moiety has the general formula:

(In the formula, R ² represents C _1-4 alkyl, and m represents an integer of 1-8.)
Item 5. The cellooligosaccharide derivative according to any one of Items 1 to 4, which is a moiety represented by:

  Item 6. The hydrophilic portion is a portion obtained by removing an anomeric hydroxyl group from a pyranose type monosaccharide and removing -H from one alcoholic hydroxyl group, or two or more selected from the group consisting of a pyranose type monosaccharide Is a moiety obtained by removing the anomeric hydroxyl group of the reducing terminal sugar residue from the 1,4-glycoside-linked oligosaccharide and removing -H from the alcoholic hydroxyl group at the 4-position of the non-reducing terminal sugar residue. The cellooligosaccharide derivative according to any one of 1 to 5.

  Item 7. Formula (I):

(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
Item 2. A cellooligosaccharide derivative according to item 1.

  Item 8. General formula (II):

(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
Item 2. A cellooligosaccharide derivative according to item 1.

  Item 9. General formula (III):

(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m ₁ and m ₂ are the same or different and are an integer of 1-8, n ₀ is an integer of 1-8. .)
Item 2. A cellooligosaccharide derivative according to item 1.

  Item 10. Formula (IV):

(In the formula, R ² represents C _1-4 alkyl, n ₁ and n ₂ are the same or different and represent an integer of 1 to 8, and m ₀ represents an integer of 1 to 8.)
Item 2. A cellooligosaccharide derivative according to item 1.

  Item 11. Item 11. A surfactant comprising the cellooligosaccharide derivative according to any one of Items 1 to 10.

  Item 12. Formula (I):

(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
The cellooligosaccharide derivative represented by the general formula (Ia):

(In the formula, R ² represents C _1-4 alkyl, and m represents an integer of 1-8.)
A method for producing a cellooligosaccharide derivative, wherein a monosaccharide or oligosaccharide having a protected hydroxyl group is bonded to a hydrophobic compound represented by the formula 1,4-glycoside, and then the protecting group is removed.

  Item 13. Item 11. A nanoparticle comprising the cellooligosaccharide derivative according to any one of Items 1 to 10.

  Item 14. Item 14. The nanoparticle according to Item 13, wherein the average particle radius is about 2.5 to 250 nm.

The present invention is described in detail below.
I. Cellooligosaccharide derivative The cellooligosaccharide derivative of the present invention has a glycosidic bond between a hydrophobic portion made of cellulose or cellooligosaccharide having a hydroxyl group alkylated and a hydrophilic portion made of a monosaccharide or oligosaccharide having a free hydroxyl group. A compound, or a mixture thereof. This cellooligosaccharide derivative is amphiphilic to water and an organic solvent, and further has a surface activity, and is used as a nonionic surfactant.

  The cellooligosaccharide derivative of the present invention is not particularly limited as long as it contains this hydrophobic part and hydrophilic part in a block form. Here, “block-like” refers to a block (hydrophobic portion) made of alkylated glucose or cellooligosaccharide, and a block (hydrophilic portion) made of monosaccharide or oligosaccharide having a free hydroxyl group in the molecule. Means that they are alternately linked in a chain.

  In addition, “alkylated glucose or cellooligosaccharide” means an alkylated one of 3 hydroxyl groups at positions 2, 3, 6 of glucose, or 2,3,6 of all glucose residues of cellooligosaccharide. Means that the three hydroxyl groups at the position are alkylated.

  Examples of the cellooligosaccharide derivative of the present invention include various cellooligosaccharide derivatives such as the following formulas.

H- (BA) x-OR ¹
HA- (BA) x-OR ^{1
R 1 - (A-B)} x-OH , or ^{R 1 -B- (A-B)} x-OH
(In the formula, A is a hydrophobic moiety obtained by removing the anomeric hydroxyl group from glucose in which the hydroxyl groups at positions 2, 3, 6 are C _1-4 alkylated and -H is removed from the alcoholic hydroxyl group at position 4. Alternatively, the anomeric hydroxyl group of the reducing terminal sugar residue is removed from the cellooligosaccharide in which the hydroxyl groups at the 2, 3, 6 positions of each glucose residue are C _1-4 alkylated and the 4th position of the non-reducing terminal sugar residue. Is a hydrophobic part obtained by removing -H from the alcoholic hydroxyl group, and B is a hydrophilic part obtained by removing the anomeric hydroxyl group from a monosaccharide and removing -H from one alcoholic hydroxyl group, or A hydrophilic moiety obtained by removing an anomeric hydroxyl group of a reducing terminal sugar residue from an oligosaccharide and removing -H from one alcoholic hydroxyl group of a non-reducing terminal sugar residue, and containing two or more A Case A is the same or different from each other May have I, when containing 2 or more B B may be the same or different, x is an integer of 1 to 20, R ¹ represents H or C _1-4 alkyl. However, sugar The chain is placed so that the reducing end is on the right side of the page.)
Examples of the cellooligosaccharide derivative of the present invention include various cellooligosaccharide derivatives such as the following formulas.

^{H-B 1} -A ¹ -OR ¹ or R ¹ -A ¹ -B ¹ -OH
(In the formula, A ¹ is a hydrophobic moiety obtained by removing the anomeric hydroxyl group from glucose in which the hydroxyl groups at the 2, 3, and 6 positions are C _1-4 alkylated and removing -H from the alcoholic hydroxyl group at the 4 position. Alternatively, the anomeric hydroxyl group of the reducing terminal sugar residue is removed from the cellooligosaccharide in which the 2, 3, 6-position hydroxyl group of each glucose residue is C _1-4 alkylated, and 4 of the non-reducing terminal sugar residue. A hydrophobic moiety obtained by removing -H from the alcoholic hydroxyl group at the position, and B ¹ is a hydrophilic moiety obtained by removing the anomeric hydroxyl group from a monosaccharide and removing -H from one alcoholic hydroxyl group; Or a hydrophilic moiety obtained by removing the anomeric hydroxyl group of the reducing terminal sugar residue from the oligosaccharide and removing -H from one alcoholic hydroxyl group of the non-reducing terminal sugar residue, and R ¹ is H or an _{C 1-4} alkyl However, the sugar chain is arranged to be the reducing end on the right side.)
As another example of the cellooligosaccharide derivative of the present invention, various cellooligosaccharide derivatives represented by the following formulas can also be mentioned.

R ¹ -A ¹ -B ¹ -A ² -OR ^{1
R 1 -A 1 -B 1 -A 2} -B 2 -OH
R ¹ -A ¹ -B ¹ -A ² -B ³ -A ³ -OR ¹
R ¹ -A ¹ -B ¹ -A ² -B ² -A ³ -B ³ -OH
^{H-B 1 -A 1 -B 2} -OH
^{H-B 1 -A 1 -B 2} -A 2 -OR 1
^{H-B 1 -A 1 -B 2} -A 2 -B 3 -OH or ^{H-B 1 -A 1 -B 2} -A 2 -B 3 -A 3 -OR 1
(In the formula, A ¹ , A ² and A ³ are the same or different and are obtained by removing the anomeric hydroxyl group from glucose in which the hydroxyl groups at the 2, 3 and 6 positions are C _1-4 alkylated and the alcoholic hydroxyl group at the 4 position. The hydrophobic moiety obtained by removing -H from the above, or the anomeric hydroxyl group of the reducing terminal sugar residue from the cellooligosaccharide in which the hydroxyl groups at the 2, 3, 6 positions of each glucose residue are C _1-4 alkylated And a hydrophobic moiety obtained by removing -H from the alcoholic hydroxyl group at the 4-position of the non-reducing terminal sugar residue, and B ¹ , B ² and B ³ are the same or different and are anomalous from a monosaccharide Hydrophilic moiety obtained by removing hydroxyl group and removing -H from one alcoholic hydroxyl group, or one of non-reducing terminal sugar residues by removing anomeric hydroxyl group of reducing terminal sugar residue from oligosaccharide -H from the alcoholic hydroxyl group of And R ¹ represents H or C _1-4 alkyl, except that the sugar chain is arranged so that the reducing end is on the right side of the page.
The hydrophobic portion is composed of glucose or cellooligosaccharide having a hydroxyl group alkylated.

  Specifically, the hydrophobic portion has the general formula:

(In the formula, R ² represents C _1-4 alkyl, and m represents an integer of 1-8.)
It is represented by

Glucose whose hydroxyl group is C _1-4 alkylated (m = 1) has a structure in which the hydroxyl groups at the 2-position, 3-position and 6-position of pyranose-type glucose are C _1-4 alkylated (particularly methylated). is doing.

The cellooligosaccharide having an alkylated hydroxyl group is composed of a cellooligosaccharide formed by linking two or more pyranose-type glucoses with 1,4-glycosides, and the hydroxyl groups at the 2nd, 3rd and 6th positions of each glucose are C. _{It has a 1-4} alkylated (particularly methylated) structure. Examples of the cellooligosaccharide include those in which 2 to 8, preferably 2 to 6, more preferably 2 to 4 glucoses are bonded.

  On the other hand, the hydrophilic portion is composed of a monosaccharide or an oligosaccharide.

  The monosaccharide is not particularly limited as long as all the hydroxyl groups except for the hydroxyl group involved in the binding with the hydrophobic moiety are free pyranose type or furanose type monosaccharides. The monosaccharide may contain a functional group other than a hydroxyl group (for example, an amino group, an acetylamino group, etc.) as long as the effect of the present invention is not adversely affected. Pyranose type monosaccharide is preferable. Specific examples of the pyranose type monosaccharide include glucose, galactose, mannose, xylose, ribose, N-acetylglucosamine, glucosamine, arabinose, rhamnose, fucose, N-acetylgalactosamine, galactosamine and the like. For example, when the monosaccharide is glucose, the hydrophilic moiety is of the formula:

It is represented by

  Examples of the oligosaccharide include oligosaccharides in which two or more selected from the group consisting of the above monosaccharides are glycoside-bonded, and preferably two or more selected from the group consisting of the above-mentioned pyranose type monosaccharide are 1,4-glycoside bonds Oligosaccharides. Specifically, cellooligosaccharide (for example, cellobiose), malto-oligosaccharide (for example, maltose, etc.), chitooligosaccharide, xylo-oligosaccharide, chitosan oligosaccharide, lactose, N-acetyllactosamine and the like can be mentioned. The oligosaccharide may be one in which 2 to 8, preferably 2 to 6, more preferably 2 to 4 monosaccharides are bonded. For example, when the oligosaccharide is cellobiose, the hydrophilic moiety is of the formula:

It is represented by

  Of the monosaccharides or oligosaccharides constituting the hydrophilic portion, glucose and cellobiose are preferable from the viewpoint of surface activity.

  As described above, the cellooligosaccharide derivative has a hydrophilic portion and a hydrophobic portion arranged in a block form, and thus has amphipathic properties and surface activity. By appropriately selecting the kind of the hydrophilic part and the hydrophobic part and the ratio of both, the cellooligosaccharide derivative of the present invention can be provided with a desired surface activity.

  Specific examples of the cellooligosaccharide derivative of the present invention include those represented by the general formula (I):

(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
Or a mixture thereof. This corresponds to a subordinate concept of the compound represented by HB ¹ -A ¹ -OR ¹ described above.

In the general formula (I), examples of the C _1-4 alkyl represented by R ¹ include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and the like. From the viewpoint of surface activity, H or a methyl group is preferable.

In the general formula (I), examples of the C _1-4 alkyl represented by R ² include methyl, ethyl, n-propyl, isopropyl, n-butyl, and isobutyl. Of these, methyl or ethyl is preferred, and methyl is particularly preferred from the standpoint of ease of synthesis and surface activity.

  In the general formula (I), m is preferably an integer of 2 to 6, and more preferably an integer of 2 to 4. Moreover, n is preferably an integer of 1 to 6, and more preferably an integer of 1 to 4.

  Further, when m is 2, n is preferably an integer of 1 to 2, when m is 3, n is preferably an integer of 1 to 3, and when m is 4, n is an integer of 1 to 4. Preferably, when m is 5, n is preferably an integer of 1 to 5, when m is 6, n is preferably an integer of 1 to 6, and when m is 7, n is 1 to 8. An integer is preferable, and when m is 8, n is preferably an integer of 1 to 8.

Specifically, in the cellooligosaccharide derivative represented by the general formula (I), the relationship between m and n is represented by the formula:
0 ≦ n ≦ 1.3m-0.6
It is particularly preferable when m is an integer of 2 to 6 and n is an integer of 1 to 6.

  Other specific examples of the cellooligosaccharide derivative of the present invention include those represented by the general formula (II):

(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
Or a mixture thereof. This corresponds to a subordinate concept of the compound represented by R ¹ -A ¹ -B ¹ -OH.

In the general formula (II), R ¹ , R ² , m and n are the same as those shown in the general formula (I).

In general formula (II), R ¹ is preferably H or a methyl group, and R ² is preferably a methyl group.

  In the general formula (II), m is preferably an integer of 2 to 6, and more preferably an integer of 2 to 4. Moreover, n is preferably an integer of 1 to 6, and more preferably an integer of 1 to 4.

  Further, when m is 2, n is preferably an integer of 1 to 2, when m is 3, n is preferably an integer of 1 to 3, and when m is 4, n is an integer of 1 to 4. Preferably, when m is 5, n is preferably an integer of 1 to 5, when m is 6, n is preferably an integer of 1 to 6, and when m is 7, n is 1 to 8. An integer is preferable, and when m is 8, n is preferably an integer of 1 to 8.

Specifically, in the cellooligosaccharide derivative represented by the general formula (II), the relationship between m and n is represented by the formula:
0 ≦ n ≦ 1.3m-0.6
It is particularly preferable when m is an integer of 2 to 6 and n is an integer of 1 to 6.

  Other specific examples of the cellooligosaccharide derivative of the present invention include those represented by the general formula (III):

(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m ₁ and m ₂ are the same or different and are an integer of 1-8, n ₀ is an integer of 1-8. .)
Or a mixture thereof. This corresponds to a subordinate concept of the compound represented by R ¹ -A ¹ -B ¹ -A ² -OR ¹ .

In the general formula (III), R ¹ and R ² are the same as those shown in the general formula (I).

In general formula (III), R ¹ is preferably H or a methyl group, and R ² is preferably a methyl group.

In the general formula (III), m ₁ is preferably an integer of ₁ to 6, and more preferably an integer of 1 to 4. m ₂ is preferably an integer of 1 to 6, an integer of 1 to 4 is more preferred. In addition, n ₀ is preferably an integer of 1 to 8, and more preferably an integer of 1 to 6.

Specifically, in the cellooligosaccharide derivative represented by the general formula (III), the relationship between m ₁ , m ₂ and n ₀ is represented by the formula:
0 ≦ n ₀ ≦ 1.3 (m ₁ + m ₂ ) −0.6
m ₁ + m ₂ is particularly preferable when it satisfies the relationship of an integer of ₂ to 8, and n ₀ is an integer of 1 to 8.

  Other specific examples of the cellooligosaccharide derivative of the present invention include those represented by the general formula (IV):

(In the formula, R ² is C _1-4 alkyl, n ₁ and n ₂ are the same or different and are an integer of 1-8, m ₀ is an integer of 1-8)
Or a mixture thereof. This corresponds to a subordinate concept of the compound represented by H-B ¹ -A ¹ -B ² —OH.

In the general formula (IV), R ¹ and R ² are the same as those shown in the general formula (I).

In the general formula (IV), n ₁ is preferably an integer of ₁ to 4, and more preferably an integer of 1 to 3. n ₂ is preferably an integer of 1 to 4, an integer of 1 to 3 is more preferable. m ₀ is preferably an integer of 2 to 8, and more preferably an integer of 2 to 6.

Specifically, in the cellooligosaccharide derivative represented by the general formula (IV), the relationship between m ₀ , n ₁ and n ₂ is represented by the formula:
0 ≦ n ₁ + n ₂ ≦ 1.3 m ₀ −0.6
It is particularly preferable that n ₁ + n ₂ satisfies an integer of 2 to 8 and m ₀ satisfies an integer of 2 to 8.

  In the cellooligosaccharide derivatives of general formulas (I) to (IV), the steric configuration at the 1-position (anomeric carbon) of each sugar may be either α- or β-. As described above, the cellooligosaccharide derivative of the present invention may be a single compound or a mixture. In the case of a mixture, the purification process of the compound is simple and the workability is excellent.

  The cellooligosaccharide derivative of the present invention does not have a long-chain hydrophobic group (C5 or more, particularly C8 or more hydrophobic group), and is composed of an oligosaccharide chain in which a hydrophilic portion and a hydrophobic portion are combined in a block manner, and water and organic It has amphiphilic properties with respect to the solvent and also has surface activity. Therefore, the cellooligosaccharide derivative of the present invention is widely useful as a nonionic surfactant.

Moreover, since the structural unit is formed from methylcellulose or an alkylated product thereof approved as a food additive, it has high safety to the human body and is biodegradable.
II. Method for producing cellooligosaccharide derivative The cellooligosaccharide derivative of the present invention can be produced, for example, as follows. These show typical examples of methods for producing cellooligosaccharide derivatives, and those skilled in the art can easily produce them with reference to these descriptions and known techniques.
II-1. Cellooligosaccharide derivative represented by general formula (I) Cellooligosaccharide derivative represented by general formula (I) is a glycoside bond of glucose or cellooligosaccharide-derived hydrophobic compound having an alcoholic hydroxyl group to a monosaccharide or oligosaccharide. Can be manufactured. If necessary, the hydroxyl group of the monosaccharide or oligosaccharide may be protected, and the protected hydroxyl group is deprotected after the glycosidic bond.

  Specifically, the cellooligosaccharide derivative represented by the general formula (I) has the general formula (Ia):

(In the formula, R ² represents C _1-4 alkyl, and m represents an integer of 1-8.)
It can manufacture by making a monosaccharide or an oligosaccharide connect with a 1, 4- glycosidic to the hydrophobic compound represented by these. The hydrophobic compound represented by the general formula (Ia) can be produced, for example, as follows (see reaction formulas 1 to 6).

  Of the general formula (Ia), the compound (9) in which m is 1 can be produced as follows.

(In the formula, R ² represents C _1-4 alkyl, and X represents a halogen atom.)
Glucose (1) is reacted with acetic anhydride to acetylate all hydroxyl groups to obtain compound (2). -OAc on the anomeric carbon of compound (2) is converted to -SPh to give compound (3), which is deacetylated to give compound (4). The diol at the 4- and 6-positions of compound (4) is protected with p-methoxybenzaldehyde dimethyl acetal to give compound (5), and R ² -X makes all free hydroxyl groups -OR ² to give compound (6). . Examples of X include iodine, bromine, and chlorine. Examples of R ² -X include methyl iodide, methyl bromide, ethyl iodide, propyl iodide and the like. Next, the compound (6) is reduced to obtain a compound (7) having a primary hydroxyl group, and this hydroxyl group is -OR ² to obtain a compound (8). The compound (9) is obtained by oxidatively removing the methoxybenzyl group from the compound (8). The hydroxyl group of compound (9) is acetylated to give compound (10), and -SPh on the 1-position (anomeric carbon) at the reducing end is substituted with -F to give compound (11). Any of these reactions can be carried out using known reactions, or can be carried out by appropriately selecting a known one from known reactions.

  In general formula (Ia), compound (20) wherein m is 2 can be produced as follows.

(In the formula, R ² represents C _1-4 alkyl, and X represents a halogen atom.)
Using cellobiose (12) as a raw material, compounds (21) and (22) are obtained according to steps a to j of Reaction Scheme 1. Any of these reactions can be carried out using known reactions, or can be carried out by appropriately selecting a known one from known reactions.

  Of the general formula (Ia), the compound (24) in which m is 3 can be produced as follows.

(In the formula, R ² represents C _1-4 alkyl.)
The compound (11) obtained by the reaction formula 1 and the compound (20) obtained by the reaction formula 2 are treated with Cp ₂ HfCl ₂ and AgClO ₄ to form a glycosidic bond to obtain a compound (23). The compound (24) is obtained by deacetylation. Any of these reactions can be carried out using known reactions, or can be carried out by appropriately selecting a known one from known reactions.

  In general formula (Ia), compound (26) wherein m is 4 can be produced as follows.

(In the formula, R ² represents C _1-4 alkyl.)
The compound (22) and the compound (20) obtained in the reaction formula 2 are treated with Cp ₂ HfCl ₂ and AgClO ₄ to form a glycoside bond to obtain a compound (25), which is deacetylated to obtain the compound ( 26) is obtained. These reactions can be carried out according to steps k to l in the above reaction scheme 3.

  In general formula (Ia), compound (28) wherein m is 5 can be produced as follows.

(In the formula, R ² represents C _1-4 alkyl.)
The compound (11) obtained in the reaction formula 1 and the compound (26) obtained in the reaction formula 4 are treated with Cp ₂ HfCl ₂ and AgClO ₄ to form glycosidic bonds to obtain a compound (27), which is removed. Compound (28) is obtained by acetylation. These reactions can be carried out according to steps k to l in the above reaction scheme 3.

  In general formula (Ia), compound (31) in which m is 6 can be produced as follows.

(In the formula, R ² represents C _1-4 alkyl.)
The compound (23) obtained by the reaction formula 3 is converted to -F using -DAST / NBS to obtain the compound (29). The compound (29) and the compound (24) obtained by the reaction formula 3 are treated with Cp ₂ HfCl ₂ and AgClO ₄ to form a glycoside bond to obtain a compound (30), which is deacetylated to obtain the compound (31) is obtained.

  Of the general formula (Ia), a compound having m = 7 and a compound having m = 8 can also be easily produced by those skilled in the art according to the above production method.

  Thus, the hydrophobic moiety (Ia) in the cellooligosaccharide derivative of the present invention can be produced. Compounds (9), (20), (24), (26), (28), (31) and the like produced by the above reaction formulas 1 to 6 are the hydrophobic compounds represented by the general formula (Ia). Equivalent to.

  Next, the cellooligosaccharide derivative of the present invention is obtained by glycosidating a monosaccharide or oligosaccharide having a hydroxyl group at the 1-position of the reducing end to the hydrophobic compound represented by the general formula (Ia). There are no particular limitations on the type of sugar used and the method of glycosidic linkage, and known methods can be employed.

  As a typical example, the hydrophobic moiety represented by the general formula (Ia) is reacted with a glucose derivative (32) to obtain a compound (Ib), which is deprotected to obtain the compound (If) of the present invention. The method is shown in Reaction Scheme 7-1.

(In the formula, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
Compound (Ia) and glucose orthoester derivative (32) are reacted in the presence of Lewis acid (BF ₃ · OEt ₂ etc.), and 3-O-benzyl-2,6-di- is added to the non-reducing end of the hydrophobic moiety. A block type cellooligosaccharide derivative (Ib) having an O-pivaloyl-D-pyranose unit is obtained.

  Since the compound (Ib) obtained in the above step m has a hydroxyl group in the molecule, it further reacts with the glucose orthoester derivative (32) present in the reaction system to introduce two or more glucopyranoside units. Blocked cellooligosaccharide derivatives are also produced. The above step m may be repeated, thereby increasing the hydrophilic pyranose unit. For example, see Example 1 (14) (15).

  Compound (Ib) is reacted with NIS / AgOTf / acetone-water to convert -SPh to -OH to obtain compound (Ic). Compound (Ic) is obtained by acetylation of hydroxyl group, removal of benzyl group, and then removal of ester with MeONa. If).

  Further, the compound (Ig) of the present invention can be obtained from the compound (Ib) obtained in the reaction formula 7-1 as shown in the reaction formula 7-2.

(In the formula, R ¹¹ is C _1-4 alkyl, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
Compound (Ib) is reacted with NIS / AgOTf / R ¹¹ —OH / CH ₂ Cl ₂ to convert —SPh to —OR ¹¹ to obtain Compound (Ic) ′, acetylation of hydroxyl group, removal of benzyl group, Subsequent ester removal with MeONa gives compound (Ig).

  As another typical example, an example in which the monosaccharide or disaccharide corresponding to the hydrophilic portion is reacted with the hydrophobic compound represented by the general formula (Ia) is shown below. First, production examples of the precursor of the hydrophilic portion are shown in Reaction Formulas 8 to 11.

  The hydroxyl group of the compound (16) obtained by the reaction formula 2 is pivaloylated to obtain a compound (33), which is treated with an acid (p-TsOH, etc.) to remove the protecting group p-methoxybenzylidene group to obtain the compound (34). Get. Only the primary hydroxyl group was converted to pivaloyl to give compound (35), and the secondary hydroxyl group was esterified with levulinic acid to obtain compound (36), and -SPh was converted to -F with DAST / NIS to give compound ( 37) is obtained.

Compound (38) was treated with dibutyltin oxide (105 ° C.), and pivaloyl chloride diluted with toluene was added dropwise to the reaction solution to which pyridine was added at −10 ° C. to give pivaloyl to obtain compound (39). The hydroxyl groups at the 1-position and 4-position are treated with acetic anhydride / pyridine and acetylated to obtain the compound (40). For example, see Journal of Wood Society 40, 302-307, Carbohydrate Research 337 (10): 951-954 (2002). This is treated with PhSH / BF ₃ .OEt ₂ to convert -OAc to -SPh to obtain compound (41). The compound (41) is treated with DBU / MeOH to obtain the compound (42). This compound (41) is treated with DAST / NBS to obtain the compound (43). To give compound -SPh as -OH (42) The compound (41) was treated with NIS / AgOTf, obtain the compound is treated with CCl ₃ CN / DBU (45) which.

  All hydroxyl groups of lactose (46) are acetylated to give compound (47), which is treated with 30% HBr / AcOH to give compound (48).

The hydroxyl group of compound (15) obtained in Reaction Scheme 2 is protected with benzaldehyde dimethyl acetal to give compound (49), and the hydroxyl group is benzylated to give compound (50). This is reduced with NaCNBH ₃ to obtain the compound (51), and the hydroxyl group is acetylated to obtain the compound (52).
DSP / NBS converts -SPh to -F to give compound (53), which is deacetylated to give compound (54). Further, the -SPh compound (52) was converted to -OH to give the compound (55) to obtain the compound is treated with CCl ₃ CN / DBU the (56) which.

  Next, the hydrophobic part represented by the general formula (Ia) and the precursor of the hydrophilic part obtained in the above reaction formulas 8 to 11 are reacted to obtain the compound represented by the general formula (I). To manufacture. The production examples are shown in the following reaction formulas 12, 13, 13 ', 14, 15 and 15'.

(In the formula, R ² represents C _1-4 alkyl.)
Compound (37) obtained in Reaction Formula 8 and compound (20) obtained in Reaction Formula 2 are glycosidically bonded to obtain Compound (57), -SPh is converted to -OH, and deesterified with sodium methoxide. To give compound (58).

(In the formula, R ² represents C _1-4 alkyl.)
The compound (43) obtained in the reaction formula 9 and the compound (20) obtained in the reaction formula 2 are glycosidically bonded to obtain a compound (59), which is reacted with NIS / AgOTf to convert -SPh to -OH. Conversion yields compound (60). Compound (60) is reduced and deesterified with sodium methoxide to give compound (61).

(In the formula, R ² represents C _1-4 alkyl.)
The compound (43) obtained by the reaction formula 9 and the compound (26) obtained by the reaction formula 4 are glycosidically bonded to obtain a compound (89), which is reacted with NIS / AgOTf to convert -SPh to -OH. Conversion yields compound (90). Compound (90) is reduced and deesterified with sodium methoxide to give compound (91).

(In the formula, R ² represents C _1-4 alkyl.)
Compound (48) obtained in Reaction Formula 10 and compound (20) obtained in Reaction Formula 2 were glycosidically bonded to obtain Compound (62), which was reacted with NIS / AgOTf to convert -SPh to -OH. Conversion yields compound (63). Compound (63) is deesterified with sodium methoxide to give compound (64).

(In the formula, R ² represents C _1-4 alkyl.)
The compound (56) obtained by the reaction formula 11 and the compound (20) obtained by the reaction formula 2 are glycosidically bonded to obtain the compound (65), which is reacted with NIS / AgOTf to convert -SPh to -OH. Conversion yields compound (66). Compound (66) is reduced and deesterified with sodium methoxide to give compound (67).

(In the formula, R ² represents C _1-4 alkyl.)
The compound (56) obtained by the reaction formula 11 and the compound (26) obtained by the reaction formula 5 are glycosidically bonded to obtain a compound (86), which is reacted with NIS / AgOTf to convert -SPh to -OH. Conversion yields compound (87). Compound (87) is reduced and deesterified with sodium methoxide to give compound (88).
II-2. Cellooligosaccharide derivative represented by general formula (II) Cellooligosaccharide derivative represented by general formula (II) is a glycoside comprising a monosaccharide or oligosaccharide having an alcoholic hydroxyl group and a hydrophobic compound derived from glucose or cellooligosaccharide. It can be manufactured by bonding. If necessary, the hydroxyl group of the monosaccharide or oligosaccharide may be protected, and the protected hydroxyl group is deprotected after the glycosidic bond.

  Typical production examples of the cellooligosaccharide derivative represented by the general formula (II) are shown in Reaction Formulas 16 to 17 below.

(In the formula, R ² represents C _1-4 alkyl.)
The compound (21) obtained in the reaction formula 2 and the compound (54) obtained in the reaction formula 11 were glycoside-bonded with NIS / AgOTf in anhydrous methylene chloride to obtain a compound (68), which was added to an aqueous solvent. Cp ₂ HfCl ₂ / AgClO ₄ is reacted to convert —F to —OH, reduced to debenzylate, and deesterified with sodium methoxide to give compound (69).

(In the formula, R ² represents C _1-4 alkyl.)
The compound (22) obtained by the reaction formula 2 and the compound (42) obtained by the reaction formula 9 are glycosidically bonded to obtain a compound (70), which is reacted with NIS / AgOTf to convert -SPh to -OH. Conversion and debenzylation by catalytic reduction and deesterification with sodium methoxide give compound (71).
II-3. A typical production example of cellooligosaccharide derivative represented by the general formula (III) in cellooligosaccharide derivative general formula represented (III), shown below in Scheme 18-19.

(In the formula, R ² represents C _1-4 alkyl.)
The compound (43) obtained in the reaction formula 9 and the compound (20) obtained in the reaction formula 2 are glycosidically bonded to obtain a compound (72), which is selectively deacetylated using DBU to obtain a compound ( 73). Compound (73) and compound (22) are glycosidically bonded to obtain compound (74), which is reacted with NIS / AgOTf to convert -SPh to -OH, debenzylation by catalytic reduction, sodium methoxide To give compound (75).

(In the formula, R ² represents C _1-4 alkyl.)
The compound (22) obtained by the reaction formula 2 and the compound (42) obtained by the reaction formula 9 are glycosidically bonded to obtain the compound (76), and this -SPh is converted to -F to convert the compound (77). obtain. On the other hand, the compound (20) obtained by the reaction formula 2 and the compound (43) obtained by the reaction formula 9 are glycoside-bonded to obtain the compound (78), which is deacetylated to obtain the compound (79). Compound (77) and compound (79) are glycosidically bonded to obtain compound (80), which is reacted with NIS / AgOTf to convert -SPh to -OH to obtain compound (81), which is obtained by catalytic reduction. Debenzylation and deesterification with sodium methoxide give compound (82).
II-4. A typical production example of cellooligosaccharide derivative represented by the general formula cellooligosaccharide derivative general formula represented by (IV) (IV), shown below in Scheme 20.

(In the formula, R ² represents C _1-4 alkyl.)
The compound (65) obtained by the reaction formula 16 is converted to -F by DAST / NBS to obtain the compound (83), and the compound (51) obtained by the reaction formula 11 is glycosidically bonded thereto. To obtain the compound (84). Compound (84) is reacted with Cp ₂ HfCl ₂ / AgClO ₄ in an aqueous solvent to convert —F to —OH, debenzylated by catalytic reduction in THF-MeOH, and deesterified with sodium methoxide. (85) is obtained.

The cellooligosaccharide derivative of the present invention can be produced from the above typical examples and using known reactions from the typical examples.
III. Features of Cellooligosaccharide Derivatives The cellooligosaccharide derivatives of the present invention are amphiphilic with respect to water and organic solvents and have surface activity. In particular, cellooligosaccharide derivatives having 5 or more sugars, preferably about 5 to 7 sugars have good surfactant ability. As a specific example, it is clear from the results of Test Examples 1 and 2.

  In addition, the cellooligosaccharide derivative of the present invention (particularly a cellooligosaccharide derivative of 5 or more sugars) is prepared by sonication (sonication time: sonication time) of an aqueous solution prepared at a critical micelle concentration (mg / ml) or higher. In an aqueous solution (about 5 to 30 seconds), rod-shaped or rugby ball-shaped nanoparticles having an average particle radius of about 2.5 to 250 nm, particularly about 5 to 150 nm are generated. The average particle radius means an average value of particle radii when the nanoparticles are observed with an atomic force microscope.

  The shape of this nanoparticle is generally a flat ellipsoidal sphere, and no change was observed in the diameter of the nanoparticle even after the drying step. Therefore, it is considered that the inside of the nanoparticle is filled with a cellooligosaccharide derivative.

  The nanoparticles composed of cellooligosaccharide derivatives having no long-chain hydrophobic group according to the present invention correspond to the partial structure of methylcellulose approved as a food additive or a pharmaceutical coating agent. Therefore, it is a highly safe and biodegradable material that is friendly to living bodies. In view of this high safety, it is useful as a drug delivery system (DDS), for example, as a nano-sized carrier (carrier) carrying a poorly water-soluble drug. Furthermore, when used for DDS applications, the desired sugar can be freely selected and introduced by tailoring the structure of the sugar moiety protruding outside the micelle, that is, by glycosylation. As a result, the drug can be transported specifically to the place where it is desired to act, and the side effects caused by the drug can be reduced, and efficient treatment can be performed.

  By the way, the inclusion function of cyclodextrin has been used so far for improving the pharmaceutical properties of pharmaceuticals and carriers for DDS, but there is no method for preparing nanoparticles comprising oligosaccharides as in the present invention.

  Thus, the hydrophobic drug can be incorporated into the nanoparticles and used as a drug carrier for inhaling the drug in the micelle sprayed in a mist to the mucous membrane and lung. Further, it can be used as a carrier for a contrast agent or a carrier for cosmetics.

  The cellooligosaccharide derivative of the present invention is composed of a sugar chain in which a hydrophobic part and a hydrophilic part are arranged in a block form, and has amphiphilic properties and surface activity, so that it can be widely used as a nonionic surfactant. . Moreover, since the raw materials are cellulose, glucose derived from starch, cellobiose and the like, and they are very easily available, they are suitable for mass production.

It is a figure which shows the process of the fractionation of the block methylated cellooligosaccharide in Test Example 1. 6 is a graph showing the measurement results of surface tension in Test Example 2. 6 is a TEM observation photograph of nanoparticles in Test Example 4. It is an AFM observation photograph of the nanoparticles in Test Example 5. It is an AFM observation photograph of the nanoparticles in Test Example 5.

  The present invention will be described in more detail with reference to examples, but is not limited thereto.

Varian Inova 300 FT-NMR (300 MHz) was used for measurement of ¹ H- and ¹³ C-NMR spectra. Bruker MALDI-TOF-MS REFLEX III was used for measurement of time-of-flight mass spectrometer (MALDI-TOF-MS) spectrum.
Example 1 (R ² = Me)
(1) Cellobiose octaacetate (13)
Cellobiose (12) (12.04 g) and sodium acetate were added to acetic anhydride (60 mL) and the mixture was stirred at 55 ° C. overnight and at 100 ° C. for 3 hours. The reaction mixture was poured into ice water (600 mL), and the organic layer was collected and concentrated. Crude crystals were filtered, washed with distilled water, and recrystallized with ethanol to obtain the title compound (20.8 g, yield 88%) as colorless crystals.
(2) Phenyl 2,3,6-tri-O-acetyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-acetyl-1-thio-β-D-glucopyranoside ( 14)
Cellobiose octaacetate (13) (20.0 g) was dissolved in anhydrous methylene chloride (70 mL), and thiophenol (2.98 mL, 0.98 equivalent) was added thereto at room temperature. Then boron trifluoride ether complex (4.46 mL, 1.2 eq) was added at 0 ° C. The reaction temperature was gradually raised to room temperature and maintained at room temperature overnight. Boron trifluoride ether complex (9 mL) was added and the reaction mixture was stirred for 1 day. The reaction mixture was diluted with ethyl acetate, washed with saturated NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give pale yellow crystals. The crude crystals were suspended in ethanol, filtered, and washed with ethanol to give the title compound (18.1 g, yield 84.0%) as colorless crystals.

¹ H-NMR (CDCl ₃ ): δ 1.9-2.2 (21H, CH ₃ CO), 3.63-3.73 (m, 2H, C _{5 '} -H, C ₅ -H), 3.75 (1H, C ₄ -H) , 4.04 (d, 1H, J _{5 ', 6'} = 2.1, J _{6 ', 6'} = 12.3, C _{6 '} -H), 4.11 (dd, 1H, J _{5, 6} = 5.4, J _{6, 6} = 11.7, C ₆ -H), 4.40 (dd, 1H, J _{5 ', 6'} = 4.2, J _{6 ', 6'} = 12.3, C _{6 '} -H), 4.49 (d, 1H, J _{1', 2 '} = 7.8, C _1' -H), 4.58 (d, 1H, J _{5, 6} = 1.8, J _{6, 6} = 11.7, C ₆ -H), 4.66 (d, 1H, J _{1, 2} = 9.9, C ₁ -H), 4.91 (t, 1H, J _{2, 3} = 9.6, C ₂ -H), 4.92 (t, 1H, J _{2 ', 3'} = 9.0, C _{2 '} -H), 5.07 (t , 1H, J _{4 ', 5'} = 9.6, C _{4 '} -H), 5.14 (t, 1H, J _{3', 4 '} = 9.3, C _3' -H), 5.19 (t, 1H, J _{3, 4} = 9.0, C ₃ -H),
¹³ C-NMR (CDCl ₃ ): δ 20.5, 20.5, 20.6, 20.7, 20.8 (CH ₃ CO), 61.4 (C-6 '), 61.9 (C-6), 67.6, 70.1, 71.5, 71.9, 72.8, 73.5, 76.3, 76.7, 85.4 (C-1), 100.7 (C-1 '), 128.3, 128.8, 131.7, 133.0 (aromatic C), 169.0, 169.2, 169.5, 169.7, 170.2, 170.4 (CH ₃ CO).
(3) Phenyl β-D-glucopyranosyl- (1 → 4) -1-thio-β-D-glucopyranoside (15)
To a solution of compound (14) (18.1 g) in tetrahydrofuran (80 mL) and methanol (20 mL) was added 28% sodium methoxide in methanol and stirred at room temperature overnight. The solution was neutralized with Dawex H ^+, filtered and Dawex H ^+, washed with methanol and 20% methanol / methylene chloride (v / v). The filtrates were combined and concentrated to dryness to give the title compound (10.8 g, yield about 100%).
(4) Phenyl 4,6-O-p-methoxybenzylidene-β-D-glucopyranosyl- (1 → 4) -1-thio-β-glucopyranoside (16)
To a suspension of compound (15) (19.5 g, 44.7 mM) and anisaldehyde dimethyl acetal (19.5 mL, 0.115 mol, 2.56 equivalent) in N, N′-dimethylformamide (50 mL) was added p-toluenesulfonic acid (6.1 g, 0.7 equivalents) was added and maintained at 45 ° C for 4 h under 25 mmHg. Then solid NaHCO ₃ (6 g) was added to the mixture and concentrated. The insoluble material was filtered and washed with 20% methanol / methylene chloride to give a syrup. The crude product was purified with a silica gel column (eluent: 10% methanol / CH ₂ Cl ₂ ) to obtain crude crystals. This was recrystallized from methanol to obtain the title compound (12.0 g, yield 48.0%) as colorless crystals.

mp 225-226 ° C; R _f value = 0.62 (10% MeOH / CH ₂ Cl ₂ , (v / v))
¹ H-NMR (CD ₃ OD / CDCl ₃ = 1/4, v / v): δ 3.33-3.40 (1H, H2), 3.40 (t, 1H, J _{2 ', 3'} = 8.7, H2 '), 3.42-3.50 (m, 1H, H5), 3.48-3.55 (m, 1H, H5 '), 3.56-3.64 (2H, H ₃ , H4), 3.66-3.76 (t, 1H, H4'), 3.70 (t , 1H, J = 9.0, H3 '), 3.7-3.8 (1H, H6), 3.81 (s, 3H, CH ₃ OPh), 3.89 (d, 2H, J _{5, 6} = 2.7, H6), 4.31 (dd , 1H, J _{5 ', 6'} = 3.6, J _{6 ', 6'} = 9.9, H6 '), 4.53 (d, 1H, J _{1', 2 '} = 7.8, H1'), 4.62 (d, 1H, J _{1, 2} = 9.9, H1), 5.51 (s, 1H, -CHPh), 7.2-7.6 (aromatic H)
¹³ C-NMR (CD ₃ OD / CDCl ₃ = 1/4, v / v): δ 54.7 (CH ₃ OPh), 60.7 (C ₆ ), 66.1, 67.7 (C _{6 '} ), 71.6 (C ₂ ), 72.6, 73.9 (C _{2 '} ), 75.8, 78.3, 79.7, 79.9, 87.5, 101.4 (-CHPh), 103.6 (C _1' ), 113.1, 127.2, 127.3, 128.5, 129.1, 131.7, 132.4, 159.7
Anal. Calcd for C ₂₆ H ₃₂ O ₁₁ S: C, 56.51; H, 5.84; Found: C, 56.11; H, 5.83
(5) Phenyl 4,6-O-p-methoxybenzylidene-2,3-di-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1 -Thio-β-D-glucopyranoside (17) (R ² = Me)
Methyl iodide (4.72 mL, 75.8 mM) and sodium hydride (1.43 g, 59.6 mM) were added to a solution of compound (16) (5.95 g, 10.8 mM) in tetrahydrofuran (40 mL) at room temperature. The reaction mixture was stirred overnight, diluted with ethyl acetate, washed with saturated aqueous NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give pale yellow crystals. The crude crystals were suspended in ethanol, filtered, and washed with ethanol to obtain the title compound (6.31 g, yield 94%) as colorless crystals.

Rf value = 0.43 (EtOAc / n-hexane = 1: 2, v / v)
mp 137-138 ℃
[α] _D ^26.3 = -43.2 ° (c 1.01, chloroform)
¹ H-NMR (CDCl ₃ ): δ 3.01 (t, 1H, J _{2 ', 3'} = 8.0, C _{2 '} -H), 3.10 (t, 1H, J _{2, 3} = 8.4, C ₂ -H) , 3.28 (t, 1H, J _{3, 4} = 8.7, C ₃ -H), 3.39, 3.58, 3.60, 3.61, 3.63 (s, 3H, C ₆ OCH ₃ , s, 3H, C _{2 '} OCH ₃ , s , 3H, C ₂ OCH ₃ , s, 3H, C ₃ OCH ₃ , s, 3H, C _{3 '} OCH ₃ ), 3.81 (s, 3H, CH ₃ OPh), 4.37 (dd, 1H, J _{5', 6 '} = 6.4 J _{6', 6 '} = 13.6, C _6' -H), 4.50 (d, 1H, J _{1 ', 2'} = 2.9, C _{1 '} -H), 4.52 (d, 1H, J _{1, 2} = 5.1, C ₁ -H), 5.50 (s, 1H, -CHPh), 7.2-7.6 (aromatic H),
¹³ C-NMR (CDCl ₃ ): δ 55.3 (CH ₃ OPh), 59.2, 60.8, 60.9, 61.1, 65.8, 68.8, 70.4, 77.5, 78.8, 81.4, 81.9, 83.2, 84.1, 86.8, 87.3, 101.1 (- CHPh), 103.5 (C _{1 '} ), 113.5, 127.3, 127.3, 128.8, 129.7, 131.8, 133.7, 160.0
Anal. Calcd for C ₃₁ H ₄₂ O ₁₁ S: C, 59.79; H, 6.80; Found: C, 59.53; H, 6.63
(6) Phenyl 4,6-O-p-methoxybenzyl-2,3-di-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1 -Thio-β-D-glucopyranoside (18) (R ² = Me)
To a solution of compound (17) (3.06 g, 4.92 mM) in anhydrous methylene chloride (6 mL) was added a THF solution (36.4 mL) of borane-tetrahydrofuran complex at −19 ° C. The temperature of the solution was gradually raised to -16 ° C over 15 minutes. After 15 min, trimethylsilyl trifluoromethanesulfonate (TMSOTf; about 540 μL) was added to the reaction mixture at −16 ° C. The temperature of the reaction mixture was gradually raised to 0 ° C. over 2 hours and maintained at 0 ° C. for 4.5 h. The mixture was cooled to −40 ° C. and triethylamine (10 mL) was added. Next, methanol was added to the solution until no more hydrogen was generated. The mixture was diluted with ethyl acetate, washed with 3N-HCl, saturated aqueous NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and concentrated to dryness to obtain crude crystals. The compound was purified by silica gel column (Wacogel C-200) eluted with ethyl acetate: n-hexane = 1: 1 (v / v) to give the title compound (2.77 g, yield 90.0%) as colorless crystals. It was.

R _f value = 0.46 (EtOAc / n-Hexane = 1: 1, v / v)
mp 94-95 ℃
[α] _D ^26.9 = -8.4 ° (c 0.771, chloroform);
¹ H-NMR (CDCl ₃ ): δ 2.93 (t, 1H, J _{2 ', 3'} = 8.0, C _{2 '} -H), 3.02 (t, 1H, J _{2, 3} = 9.6, C ₂ -H) , 3.23 (t, 2H, J _{3, 4} = 8.7, J _{3 ', 4'} = 8.7, C ₃ -H, C _{3 '} -H), 3.34, 3.53, 3.57, 3.57, 3.63 (s, 3H, C ₆ OCH ₃ , s, 3H, C _{2 '} OCH ₃ , s, 3H, C ₂ OCH ₃ , s, 3H, C ₃ OCH ₃ , s, 3H, C _3' OCH ₃ ), 3.78 (s, 3H, CH ₃ OPh), 4.37 (dd, 1H, J _{5 ', 6'} = 6.4, J _{6 ', 6'} = 13.6, C _{6 '} -H), 4.31 (d, 1H, J _{1', 2 '} = 7.6, C _{1 '} -H), 4.47 (d, 1H, J _{1, 2} = 10.0, C ₁ -H), 4.51, 4.72 (d, 2H, 2H, J _gem = 10.8, 11.0, -CH ₂ Ph), 7.2 -7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 55.3 (CH ₃ OPh), 59.1, 60.6, 60.9, 61.1, 62.6 (C _{6 '} ), 70.5, 74.4, 74.6, 77.7, 79.9, 82.2, 84.3, 86.4, 87.0, 87.3, 102.6, 113.9, 127.4, 128.8, 129.8, 130.1, 131.8, 133.7, 159.4
Anal.Calcd for C ₃₁ H ₄₄ O ₁₁ S: C, 59.60; H, 7.10; Found: C, 59.46; H, 7.07
(7) Phenyl 4-O-p-methoxybenzyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1 -Thio-β-D-glucopyranoside (19) (R ² = Me)
To a solution of compound (18) (6.66 g, 10.7 mM) in tetrahydrofuran (30 mL) at 0 ° C., sodium hydride (60% in mineral oil, 0.512 g) and methyl iodide (0.8 mL, 12.8 mM) were added. It was. The reaction mixture was maintained at room temperature for 5 h, and the mixture was diluted with ethyl acetate, washed with saturated aqueous NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and concentrated to dryness to obtain crude crystals. The crude crystals were recrystallized with ethanol to obtain the title compound (5.78 g, yield 84.9%) as colorless crystals.

Rf value = 0.70 (EtOAc / n-hexane = 1: 1, v / v)
mp 82-84 ℃
[α] _D ^27.3 = -22.7 ° (c 0.998, chloroform)
¹ H-NMR (CDCl ₃ ): δ 2.93 (t, 1H, J _{2 ', 3'} = 8.0, C _{2 '} -H), 3.06 (t, 1H, J _{2, 3} = 9.6, C ₂ -H) , 3.19 (t, 1H, J _{3 ', 4'} = 9.1, C _{3 '} -H), 3.27 (t, 1H, J _{3, 4} = 8.7, C ₃ -H), 3.34, 3.36, 3.52, 3.57, 3.59, 3.62 (s, 3H, C ₆ OCH ₃ , s, 3H, C _{6 '} OCH ₃ , s, 3H, C _2' OCH ₃ , s, 3H, C ₂ OCH ₃ , s, 3H, C ₃ OCH ₃ , s, 3H, C _{3 '} OCH ₃ ), 3.78 (s, 3H, CH ₃ OPh), 4.29 (d, 1H, J _{1', 2 '} = 8.0, C _1' -H), 4.48 (d, 1H , J _{1, 2} = 9.6, C ₁ -H), 4.53, 4.72 (d, 2H, 2H, J _gem = 10.4, 10.4, -CH ₂ Ph), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 55.2, 59.1, 59.3, 60.6, 60.6, 60.7, 61.0, 70.5, 71.1, 74.5, 74.6, 77.1, 77.7, 79.0, 81.9, 84.2, 86.7, 87.0, 87.3, 103.3, 113.8, 127.3, 128.8, 129.7, 130.4, 131.7, 133.9, 159.3
(8) Phenyl 2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D-glucopyranoside ( 20) (R ² = Me)
To a solution of compound (19) (2.00 g, 3.21 mM) in acetonitrile: water = 9: 1 (v / v) (66.7 mL) at 0 ° C., ammonium cerium (IV) nitrate (3.52 g, 6.41 mM) was added. It was. The reaction mixture was maintained at 0 ° C. for 12 h. The mixture was diluted with ethyl acetate, washed with saturated aqueous NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and concentrated to dryness to obtain crude crystals. The crude product was purified by silica gel column eluting with methylene chloride, ethyl acetate: n-hexane = 1: 1 (v / v) and ethyl acetate to give the title compound as colorless crystals (1.52 g, yield 91.4%) Got.

mp 96-97 ℃
[α] _D ^27.5 = -47.0 ° (c 1.03, chloroform)
Rf value = 0.17 (EtOAc / n-Hexane)
¹ H-NMR (CDCl ₃ ): δ 2.93 (t, 1H, J _{2 ', 3'} = 8.0, C _{2 '} -H), 3.06 (t, 1H, J _{2, 3} = 9.2, C ₂ -H) , 3.08 (t, 1H, J _{3 ', 4'} = 9.2, C _{3 '} -H), 3.25 (t, 1H, J _{3, 4} = 8.7, C ₃ -H), 3.36, 3.38, 3.51, 3.57, 3.57, 3.61 (s, 3H, C ₆ OCH ₃ , s, 3H, C _{6 '} OCH ₃ , s, 3H, C _2' OCH ₃ , s, 3H, C ₂ OCH ₃ , s, 3H, C ₃ OCH ₃ , s, 3H, C _{3 '} OCH ₃ ), 4.34 (d, 1H, J _{1', 2 '} = 7.8, C _1' -H), 4.48 (d, 1H, J _{1, 2} = 9.8, C _1- H), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 59.1, 59.5, 60.4, 60.5, 60.6, 60.8, 70.4, 71.7 (C _{4 '} ), 72.8, 73.4 (C _5' ), 77.5 (C ₄ ), 78.9 (C ₅ ), 81.8 (C ₂ ), 83.7 (C _{2 '} ), 86.1 (C _3' ), 86.6 (C ₃ ), 87.2 (C ₁ ), 103.2 (C _{1 '} ), 127.3, 128.7, 131.7, 133.8
Anal. Calcd for C ₂₄ H ₃₈ O ₁₀ S: C, 55.58; H, 7.39; Found: C, 55.50; H, 7.24
(9) Phenyl 4-O-acetyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio- β-D-glucopyranoside (21) (R ² = Me)
Acetic anhydride (320 μL) and pyridine (275 μL) were added to a solution of compound (20) (800 mg, 1.54 mM) in methylene chloride (5 mL), and the mixture was stirred overnight. The mixture was diluted with ethyl acetate, washed with 1N-HCl and saturated aqueous NaHCO ₃ and brine, dried over Na ₂ SO ₄ and concentrated to dryness to quantitatively obtain the title compound as colorless crystals.

mp 94.8-95.7 ° C
[α] _D ^27.2 = -37.8 ° (c 3.76, chloroform)
¹ H-NMR (CDCl ₃ ): δ 2.09 (COCH3), J _{2 ', 3'} = 9.3, C _{2 '} -H), 3.08 (dd, 1H, J _{2, 3} = 9.0, C ₂ -H), 3.24 (t, 1H, J _{3 ', 4'} = 9.0, C _{3 '} -H), 3.28 (t, 1H, J _{3, 4} = 8.7, C ₃ -H), 3.34, 3.39, 3.51, 3.54, 3.59 , 3.60 (s, 3H, C ₆ OCH ₃ , s, 3H, C _{6 '} OCH ₃ , s, 3H, C _2' OCH ₃ , s, 3H, C ₂ OCH ₃ , s, 3H, C ₃ OCH ₃ , s, 3H, C _{3 '} OCH ₃ ), 4.38 (d, 1H, J _{1', 2 '} = 7.8, C _1' -H), 4.51 (d, 1H, J _{1, 2} = 9.9, C ₁ -H ), 4.87 (t, 1H, J _{4 ', 5'} = 9.6, C _{4 '} -H), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.8 (Ac), 59.1, 59.4, 60.3, 60.6, 60.6, 60.7, 70.3, 70.8 (C _{4 '} ), 72.0, 72.9 (C _5' ), 77.6 (C ₄ ) , 78.8, 81.7 (C ₂ ), 83.5 (C _{2 '} ), 83.9 (C _3' ), 86.5 (C ₃ ), 87.2 (C ₁ ), 103.0 (C _{1 '} ), 127.3, 128.7, 131.7, 133.8, 169.8
(10) 4-O-acetyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-D-glucopyranosyl fluoride ( 22) (R ² = Me)
To a solution of compound (21) (864 mg, 1.54 mM) in anhydrous methylene chloride (20 mL) at −78 ° C., (diethylamino) sulfur trifluoride (DAST) (612 μL, 4.63 mM, 3.0 equivalents) and N-bromosuccinimide ( NBS) (320 mg, 1.80 mM, 1.17 equiv) was added. The temperature of the reaction mixture was gradually raised to -25 ° C over 11h and the solution was maintained at -25 ° C for 9h. The mixture was diluted with ethyl acetate, washed with saturated aqueous NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give a colorless oil quantitatively. The crude product was purified by silica gel column (Wacogel C-200) eluting with ethyl acetate: n-hexane = 1: 2 (v / v) to give the title compound as colorless crystals (725 mg, about 100% yield) Got. α: β = 4: 1 (calculated from ¹ H-NMR data).
mp 56.0-63.5 ℃
[α] _D ^27.7 = + 27.7 ° (c 3.76, chloroform)
¹ H-NMR (CDCl ₃ ): δ 2.06 (s, 3H, CH ₃ CO), 3.05 (t, 1H, J _{2 ', 3'} = 9.3, C _{2 '} -H), 3.16-3.32 (1H, C ₂ -H), 3.25 (t, 1H, J _{3 ', 4'} = 9.6, C _{3 '} -H), 3.50-3.62 (1H, C ₃ -H), 3.33, 3.38, 3.50, 3.52, 3.54, 3.57 (s, 3H, C ₆ OCH ₃ , s, 3H, C _{6 '} OCH ₃ , s, 3H, C _2' OCH ₃ , s, 3H, C ₂ OCH ₃ , s, 3H, C ₃ OCH ₃ , s, 3H, C _{3 '} OCH ₃ ), 4.35 (d, 1H, J _{1', 2 '} = 7.8, C _1' -H), 4.86 (t, 1H, J _{4 ', 5'} = 7.0, C _{4 '} - H), 5.16 (dd, 1H, J _{1β, 2} = 6.3, J _{1β, F} = 53.1, C ₁ -H), 5.65 (dd, 1H, J _{1α, 2} = 2.7, J _{1α, F} = 53.4, C ₁ -H)
¹³ C-NMR (CDCl ₃ ): δ 20.9, 58.9, 59.1, 59.2, 59.5, 60.1, 60.3, 60.7, 60.7, 69.4 (C ₆ ), 70.8 (C _{4 '} ), 72.0 (C _6' ), 72.3 ( C ₅ ), 72.3 (C ₅ ), 73.1 (C _{5 '} ), 76.7 (C ₄ ), 76.8 (C ₄ ), 80.2 (C ₂ ), 80.6 (C ₂ ), 80.8 (C ₃ ), 83.5 (C _{2 '} ), 83.9 (C _3' ), 103.0, 103.1 (C _{1 '} ), 103.4 (C _1α ), 106.4 (C _1α ), 169.9 (CH3CO)
MALDI-TOF-MS found [M + Na] ⁺ = 493.15
(11) Compounds (2) to (11) represented by reaction formula 1
Compounds (2) to (11) represented by Reaction Formula 1 can be produced according to the production methods of Compounds (13) to (22) represented by Reaction Formula 2 shown in Examples 1 (1) to (10). it can.
(12) General method of alkylated cellooligosaccharide chain formation Compound (20) obtained in (8) above (69.6 mg, 0.134 mM) and compound (22) obtained in (10) above (70.2 mg, 0.149) mM) was dried overnight under reduced pressure, and then compound (20) and compound (22) were dissolved in anhydrous benzene (5 mL). The reaction mixture was stirred for 2 h at room temperature with active molecular sieve 4A (500 mg) under a nitrogen atmosphere. To the solution of compound (20) and compound (22), Cp ₂ HfCl ₂ (62.3 mg, 0.164 mM) and AgClO ₄ (68.1 mg, 0.328 mM) were added at room temperature, and the mixture was continuously stirred for 1 h. The insoluble material was removed by filtration and washed with ethyl acetate. The filtrate was diluted with ethyl acetate, washed with distilled water, aqueous NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The product was isolated by preparative TLC (hereinafter referred to as PTLC) (eluent: EtOAc: n-hexane = 1: 1, 1: 2 (v / v)) to obtain the starting compound (20) (15.0 mg ), Compound (25) (28.0 mg) and compound (25α) (40.8 mg) were obtained.
Phenyl 4-O-acetyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- ( 1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D-glucopyranoside (25) (R ² = Me)
[α] _D ^28.8 = -15.7 ° (c 0.942, chloroform)
¹ H-NMR (CDCl ₃ ): δ 2.09 (COCH ₃ ), 2.96, 2.96 (t, t, 1H, 1H, J = 9.0, C _{2 '} -H or C _2'' -H), 3.02 (t, 1H, J _{2 ''',3'''} = 9.3, C _{2 '''} -H), 3.09 (t, 1H, J _{2, 3} = 9.0, C ₂ -H), 3.21 (t, 2H, J = 9.0, C _{3 '} -H and C _3'' -H), 3.24 (t, 1H, J _{3''', 4 '''} = 9.0, C _3''' -H), 3.28 (t, 1H _{, J 3, 4 = 9.0,} C 3 -H), 3.24 - 3.32 (C 5 '-H and C 5''-H), 3.32 - 3.38 (C 5 -H), 3.42 - 3.49 (m, 1H, C _{5 '''} -H), 3.68 (2H, J = 9.0, C _4' -H and C _{4 ''} -H), 3.71 (t, 1H, J _{4, 5} = 9.3, C ₄ -H), 4.32 (d. 1H, J = 7.8, C 1 '-H or C 1''-H), 4.34 (d. 1H, J = 8.4, C 1' -H or C 1 '' -H), 4.41 ( d, 1H, J _{1 ''',2'''} = 7.8, C _{1 '''} -H), 4.51 (d, 1H, J _{1, 2} = 9.9, C ₁ -H), 4.87 (t, 1H , J _{4 ''',5'''} = 9.6, C _{4 '''} -H), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.9, 59.1, 59.4, 60.1, 60.3, 60.4, 60.5, 60.6, 60.6, 60.7, 60.8, 70.1 (C ₆ or C _{6 '} or C _6'' or C _6''' ), 70.4 (C ₆ or C _6' or C _{6 ''} or C _{6 '''} ), 70.9 (C _4''' ), 72.0 (C ₆ or C _{6 '} or C _6'' or C _6''' ), 72.9 (C _{5 '''} ), 74.7 (C _5' or C _{5 ''} ), 74.8 (C _{5 '} or C _5'' ), 77.3 (C ₄ or C _4' or C _{4 ''} ), 77.4 (C ₄ or C _{4 '} or C _4'' ), 77.8 (C _4' or C _{4 ''} ), 78.9 (C ₅ ), 81.9 (C ₂ ), 83.2 (C _{2 '} or C _2'' or C _2''' ), 83.4 (C _{2 '} or C _2'' or C _2''' ), 83.5 (C _{2 '} or C _2'' or C _2''' ), 83.9 (C _{3 '''} ), 84.8 (C _{3 '} or C _3'' ), 85.0 (C _3' or C _{3 ''} ), 86.7 (C ₃ ), 87.2 (C ₁ ), 102.9 (C _{1 '} or C _1'' or C _{1 '''} ), 103.2 (C _1' or C _{1 ''} or C _{1 '''} ), 103.3 (C _1' or C _{1 ''} or C _{1 '''} ), 127.2, 128.7, 131.7 , 133.8 (aromatic C), 169.9 (CH ₃ CO)
ESI in methanol [M + Na] ⁺ = 991, [M + K] ⁺ = 1007
MALDI-TOF-MS [M + Na] ⁺ = 991.18, [M + K] ⁺ = 1007.12
Phenyl 4-O-acetyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-α-D-glucopyranosyl- ( 1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D-glucopyranoside (25α) (R ² = Me)
[α] _D ^29.2 = + 26.2 ° (c 2.67, chloroform)
¹ H-NMR (CDCl ₃ ): δ 2.09 (COCH ₃ ), 3.33, 3.35, 3.37, 3.38, 3.52, 3.52, 3.54, 3.55, 3.57, 3.57, 3.58, 3.60, 3.61 (OCH ₃ ), 4.50 (d, 1H, J _{1, 2} = 9.8, H1), 4.90 (t, 1H, J _{4 ''',5'''} = 9.2, H4 '''), 5.67 (d. 1H, J = 3.8, H1'' ), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.9, 59.0, 59.1, 59.5, 60.3, 60.4, 60.5, 60.6, 60.7, 68.2, 69.8, 70.4, 70.6, 71.1, 72.1, 73.1, 73.9, 79.0, 81.1, 81.3, 81.9, 83.6, 84.2, 84.3, 86.7, 86.9, 87.4, 95.8, 103.9, 127.3, 128.8, 131.9, 133.9, 169.9;
ESI in methanol [M + Na] ⁺ = 991.2, [M + K] ⁺ = 1007.1
Phenyl 4-O-acetyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- ( 1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-α-D-glucopyranoside (25 ') (R ² = Me)
¹ H-NMR (CDCl ₃ ): δ 2.10 (COCH ₃ ), 3.35, 3.38, 3.41, 3.52, 3.52, 3.53, 3.56, 3.58, 3.59, 3.61, 3.62 (OCH ₃ ), 4,32 (d, 1H, J = 7.8), 4.37 (d, 1H, J = 7.8), 4.42 (d, 1H, J = 7.8), 4.90 (t, 1H, J _{4 ''',5'''} = 9.3, H4 ''' ), 5.75 (d. 1H, J = 5.1, H ₁ ) 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.9, 58.1, 58.9, 59.1, 59.2, 59.4, 60.2, 60.3, 60.6, 60.6, 60.7, 60.8, 69.9, 70.1, 70.9, 72.1, 72.9, 73.0, 74.7, 74.8, 77.4, 77.6, 80.8, 81.8, 83.3, 83.4, 83.5, 83.9, 84.8, 85.0, 86.3, 102.9, 103.2, 103.4, 126.9, 128.9, 130.8, 134.6, 169.9
Phenyl 4-O-acetyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-α-D-glucopyranosyl- ( 1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-α-D-glucopyranoside (25α ') (R ² = Me)
¹ H-NMR (CDCl ₃ ): δ 2.10 (COCH ₃ ), 3.34, 3.36, 3.37, 3.40, 3.51, 3.52, 3.55, 3.56, 3.57, 3.58, 3.58, 3.62 (OCH ₃ ), 4.33 (d, 1H, J = 7.8), 4.36 (d, 1H, J = 7.8), 4.90 (t, 1H, J _{4 ''',5'''} = 9.3, H4 '''), 5.69 (d. 1H, J = 3.6 , H1``), 5.76 (d, 1H, J _{1, 2} = 5.1, H1)
¹³ C-NMR (CDCl ₃ ): δ 20.9, 58.1, 58.9, 59.1, 59.4, 59.5, 59.5, 60.3, 60.5, 60.6, 60.8, 69.6, 69.8, 70.4, 70.9, 72.0, 73.0, 73.8, 77.7, 80.7, 80.9, 81.2, 81.7, 83.4, 84.1, 86.3, 86.9, 95.6, 103.2, 103.3, 126.9, 128.9, 130.7, 134.6, 169.9
(13) Phenyl 2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4 ) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D-glucopyranoside (26) (R ² = Me)
To a solution of compound (25) (29.7 mg, 30.6 μM) in tetrahydrofuran (1 mL) and methanol (0.1 mL) was added 28% sodium methoxide in methanol (4 μL, 70 μM) at room temperature, and the mixture was stirred for 30 minutes. The reaction mixture was diluted with ethyl acetate, washed with water and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give compound (26) (28.4 mg, about 100% yield).

mp 144-145 ° C;
[α] _D ^29.5 = -16.0 ° (c 1.59, chloroform)
¹ H-NMR (CDCl ₃ ): δ 2.92 -3.02 (dd, J = 8.1, 9.0, dd, J = 8.1, 9.3, 3H, C _{2 '} -H, C _2'' -H, and C _2''' -H), 3.09 (t, 1H, J _{2, 3} = 9.0, C ₂ -H), 3.12 (t, 1H, J _{3''', 4 '''} = 8.1, C _3''' -H ), 3.22 (t, 2H, J = 9.0, C _{3 '} -H and C _3'' -H), 3.29 (t, 1H, J _{3, 4} = 8.4, C ₃ -H), 3.33-3.39 (m , 1H, C _{5 '''} -H), 3.35-3.40 (m, 1H, C ₅ -H), 3.53-3.56 (1H, C _4''' -H), 3.66-3.74 (C ₄ -H, C _{4 '} -H, and C _4'' -H), 3.62-3.84 (C ₆ -H, C _6' -H, C _{6 ''} -H, C _{6 '''} -H), 4.32 (d. 1H, J = 8.7, C _{1 '} -H or C _1'' -H), 4.35 (d. 1H, J = 7.8, C _1' -H or C _{1 ''} -H), 4.40 (d, 1H, J _{1 ''',2'''} = 7.8, C _{1 '''} -H), 4.51 (d, 1H, J _{1, 2} = 9.9, C ₁ -H), 3.38, 3.39, 3.40, 3.40, 3.54 , 3.56, 3.59, 3.60, 3.61, 3.64 (s, 3H, 3H, 3H, 3H, 9H, 3H, 3H, 3H, 3H, 3H, OCH ₃ ), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 59.1, 59.6, 60.0, 60.4, 60.4, 60.6, 60.6, 60.7, 60.8, 60.8, 70.1 (C ₆ or C _{6 '} or C _6'' or C _6''' ) , 70.4 (C ₆ or C _{6 '} or C _6'' or C _6''' ), 72.0 (C _{4 '''} ), 73.0 (C ₆ or C _6' or C _{6 ''} or C _{6 '''} ), 73.1 (C _{5 '''} ), 74.7 (C _5' and C _{5 ''} ), 77.1 (C ₄ or C _{4 '} or C _4'' ), 77.8 (C ₄ or C _4' or C _{4 ''} ), 78.9 (C ₅ ), 81.9 (C ₂ ), 83.2 (C _2' or C _{2 ''} or C _{2 '''} ), 83.4 (C _2' or C _{2 ''} or C _{2 '''} ) , 83.6 (C _{2 '} or C _2'' or C _2''' ), 84.8 (C _{3 '} or C _3'' ), 85.0 (C _3' or C _{3 ''} ), 86.0 (C _{3 '''} ), 86.7 (C ₃ ), 87.3 (C ₁ ), 103.1 (C _{1 '} or C _1'' or C _1''' ), 103.2 (C _{1 '} or C _1'' or C _1''' ), 103.3 (C _{1 '} or C _1'' or C _1''' ), 127.3, 128.8, 131.7, 133.9 (aromatic C)
MALDI-TOF-MS (Positive Reflector mode): calculated m / z 926.42; found m / z: [M + Na] ⁺ = 949.34, [M + K] ⁺ = 965.32
(14) 3-O-benzyl-6-O-pivaloyl-α-D-glucopyranose 1,2,4-orthopivalate (32) and compound (26) [(Ia) (R ² = Me m = 4)] All of the reaction reactions were carried out using a high vacuum line capable of maintaining a vacuum of 1 × 10 ⁻³ Torr. Compound (26) (18 mg, 19.4 μM) and compound (32) (16.3 mg, 38.9 μM) were dried together with active molecular sieve 4A (200 mg) in a glass reaction vessel under reduced pressure for about 1 day. Distilling methylene chloride from CaH _2, it was degassed three times freezing and thawing in a high vacuum line, was transferred into the reaction vessel. BF ₃ · Et ₂ O was added to the reaction vessel with a syringe through a rubber septum. Thereafter, the glass reaction vessel was burned and separated with a gas burner, transferred to a bath at an appropriate temperature, and stirred continuously for 19 hours. To the reaction mixture was added triethylamine (8.1 mL, 58 μM), filtered and washed with ethyl acetate. The combined filtrate was washed with distilled water and brine, dried over anhydrous sodium sulfate, and concentrated to dryness. In the obtained mixture, the raw material tetramer (59%), pentamer (32%), hexamer (10%), heptamer (1%) and octamer (1%) were obtained.

  For convenience, tetramer is a compound with 4 monosaccharides (n = 0), pentamer is a compound with 5 monosaccharides (n = 1), and hexamer is 6 monosaccharides. The compound (n = 2), the heptamer is used to mean a compound (n = 3) in which seven monosaccharides are bonded, and the octamer is used to mean a compound (n = 4) in which eight monosaccharides are bonded.

  Thus, in the above reaction, not only one glucose unit but also two or more glucose units can be bonded (polymerized) to the cellotetramer of the raw material compound (26).

The spectral data of each product (Ib) is shown below.
Phenyl 3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-Tri-O-methyl-1-thio-β-D-glucopyranoside (Ib) (m = 4, n = 1)
¹ H-NMR (CDCl ₃ ): δ 1.84, 1.21 (s, s, 9H, 9H, COC (CH ₃ ) ₃ ), 2.86-3.00 (3H, C _{2 '} -H, C _2''' -H) , 3.07 (1H, C ₂ -H), 3.78, 3.40, 3.42, 3.53, 3.54, 3.57, 3.58, 3.58, 3.60, 3.61 (OCH ₃ ), 4.22 (dd, 1H, J = 2.0, J = 12.3, C 6`` <sub>''</sub> -H), 4.29, 4.31 (1H, 1H, J = 6.9, J = 7.8, C <sub>1 '</sub> -H or C <sub>1''</sub> -H), 4.34 (d, 1H, J = 8.1, C <sub>1 '''</sub> -H), 4.51 (d, 1H, J = 9.9, C <sub>1</sub> -H), 4.57 (d, 1H, J = 8.1, C 1`` _'' -H), 4.61 (dd, 1H, J = 3.0, J = 12.3, C 6`` <sub>''</sub> -H), 4.82 (d, 1H, J = 11.4, CH <sub>2</sub> Ph), 4.68 (d, 1H, J = 11.0, CH <sub>2</sub> Ph), 4.97 ( t, 1H, J = 8.4, C 2`` _'' -H), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 27.2 (COC (CH ₃ ) ₃ ), 27.2 (COC (CH ₃ ) ₃ ), 38.8 (COC (CH ₃ ) ₃ ), 39.0 (COC (CH ₃ ) ₃ ), 59.1, 59.4, 60.4, 60.6, 60.7, 60.9, 62.6, 70.1, 72.9, 74.0, 74.6, 747, 74.8, 75.8, 77.2, 77.8, 78.9, 81.9, 82.4, 83.3, 83.4, 84.9, 86.7, 87.3, 100.0 ( C _{1 ''''} ), 103.2, 103.3, 127.3, 127.7, 128.5, 128.8, 131.7, 133.9, 138.1, 176.8 (COC (CH ₃ ) ₃ ), 179.8 (COC (CH ₃ ) ₃ )
MALDI-TOF-MS (Positive Reflector mode): calculated m / z 1346.63; found m / z: [M + Na] ⁺ = 1369.33
Phenyl 3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl- (1 → 4) -3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl -(1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- ( 1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D-glucopyranoside (Ib) (m = 4, n = 2)
¹ H-NMR (CDCl ₃ ): δ 1.04, 1.05, 1.21, 1.24 (s, s, s, s, 9H, 9H, 9H, 9H, COC (CH ₃ ) ₃ ), 2.82-3.01 (3H, C _{2 '} -H, C _2'' -H, C _2''' -H), 3.37, 3.38, 3.38, 3.40, 3.51, 3.53, 3.56, 3.58, 3.60, 3.61 (OCH ₃ ), 3.97 (t, 1H, J = 9.3, C 4`` _'' -H), 4.18-4.24 (2H, C _{6 '''''} -H), 4.27, 4.31, 4.33 (d, d, d, 1H, 1H, 1H, J = 7.8, J = 7.5, J = 7.2, C _{1 '} -H or C _1'' -H or C _1''' -H), 4.33-4.38 (2H, C _{6 ''''} -H), 4.42 (t, 1H, J = 8.1, C _{1 '''''} -H), 4.48 (d, 1H, J = 11.4, CH ₂ Ph), 4.48-4.54 (d, 1H, C _1'''' - H), 4.51 (d, 1H, J = 9.9, C ₁ -H), 4.48 (d, 1H, J = 11.4, CH ₂ Ph), 4.76 (d, 1H, J = 11.7, CH ₂ Ph), 5.01 (t, 1H, J = 8.1, C _{2 '''''} -H), 4.91 (t, 1H, J = 8.7, C _2'''' -H), 5.11 (d, 1H, J = 11.4, CH ₂ Ph), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 27.0 (COC (CH ₃ ) ₃ ), 27.0 (COC (CH ₃ ) ₃ ), 27.2 (COC (CH ₃ ) ₃ ), 27.2 (COC (CH ₃ ) ₃ ), 38.6 (COC (CH ₃ ) ₃ ), 38.7 (COC (CH ₃ ) ₃ ), 38.9 (COC (CH ₃ ) ₃ ), 38.9 (COC (CH ₃ ) ₃ ), 59.1, 59.1, 59.4, 60.4, 60.6, 60.7, 60.8, 60.8, 60.9, 63.0, 70.1, 70.5, 72.3, 72.9, 74.2, 74.9, 76.2, 77.2, 78.9, 81.2, 81.9, 82.4, 83.4, 84.7, 85.0, 86.7, 87.3, 100.0, 100.2, 103.1, 126.8, 127.4, 127.9, 128.0, 128.5, 128.8, 130.9, 131.7, 133.9, 137.9, 138.8, 176.7 (COC (CH ₃ ) ₃ ), 176.8 (COC (CH ₃ ) ₃ ), 177.9 (COC (CH ₃ ) ₃ ), 179.4 (COC (CH ₃ ) ₃ )
MALDI-TOF-MS (Positive Reflector mode): calculated m / z 1766.85; found m / z: [M + Na] ⁺ = 1790.00
Phenyl 3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl- (1 → 4) -3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl -(1 → 4) -3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D -Glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl -(1 → 4) -2,3,6-Tri-O-methyl-1-thio-β-D-glucopyranoside (Ib) (m = 4, n = 3)
MALDI-TOF-MS (Positive Linear mode): calculated m / z 2188.55; found m / z: [M + Na] ⁺ = 2211.86
Phenyl 3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl- (1 → 4) -3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl -(1 → 4) -3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl- (1 → 4) -3-O-benzyl-2,6-di-O-pivaloyl -β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β -D-Glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio -β-D-Glucopyranoside (Ib) (m = 4, n = 4)
MALDI-TOF-MS (Positive Linear mode): calculated m / z 2609.04; found m / z: [M + Na] ⁺ = 2632.29
(15) Examination of polymerization method of compound (26) and 3-O-benzyl-6-O-pivaloyl-α-D-glucopyranose 1,2,4-orthopivalate (32) Reaction product of (134) above Was repeatedly subjected to the above step (13) to examine the distribution ratio of the polymerization reaction product. As can be seen from Table 1, it was confirmed that by repeating the step (13), the number of compounds having an increased degree of polymerization of glucose units increased.

(16) General Deprotection Method for Library (Ib) Compound (Ib) (25.6 mg, 19.4 μM) in a methylene chloride (5 mL) solution containing a small amount of MeOH and water was mixed with NIS (6.1 mg, 27.2 μM). ) And a catalytic amount of AgOTf were added at room temperature and stirred continuously for 3 h at room temperature. The reaction mixture was diluted with ethyl acetate, washed with aqueous NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give a crude product. The crude product was purified by column chromatography (eluent: ethyl acetate: n-hexane = 1: 4, v / v) to obtain compounds (Ic) and (Ic) ′ (25.3 mg).

  Compounds (Ic) and (Ic) 'were acetylated with acetic anhydride (1 mL) and pyridine (1 mL) at room temperature overnight to obtain compounds (Id) and (Id)'.

  To a solution of compounds (Id) and (Icd) 'in tetrahydrofuran (1 mL) and ethanol (1 mL) was added palladium hydroxide / carbon (80 mg), and the mixture was stirred at room temperature for 11 h in a hydrogen atmosphere. Palladium hydroxide / carbon was filtered off and washed with 20% methanol / methylene chloride (v / v). The combined filtrates were concentrated and dried to obtain compounds (Ie) and (Ie) ′ (21.8 mg).

  To a solution of compounds (Ie) and (Ie) 'in tetrahydrofuran (4 mL) and methanol (1 mL) was added 28% sodium methoxide (0.1 mL), and the mixture was stirred at room temperature overnight. The mixture was neutralized with Dawex H +, Dawex H + was filtered off and washed with 20% methanol / methylene chloride (v / v) and methanol. The combined filtrate was concentrated and dried to obtain compounds (If) and (Ig) (19.0 mg).

Compound (Ig):
¹ H-NMR (CDCl ₃ ): δ 2.96 (C ₂ -H (methylated glucose)), 3.40, 3.54, 3.57, 3.58, 3.59 (OCH ₃ ), 4.33 (C ₁ -H (methylated glucose), 4.58 (C ₁ -H (non-methylated glucose)); ¹ H-NMR (D ₂ O at 20 ° C): δ 3.14 (C ₂ -H (methylated glucose)), 3.41, 3.58, 3.59 (OCH ₃ ), 4.43 ( C ₁ -H (methylated glucose), 4.45 (C ₁ -H (non-methylated glucose)); ¹ H-NMR (CD ₃ OD): δ 2.90 (C ₂ -H (methylated glucose)), 3.40, 3.53, 3.56, 3.58 (OCH ₃ ), 4.38 (C ₁ -H (methylated glucose), (C ₁ -H (non-methylated glucose)); ¹³ C-NMR (CDCl ₃ ): δ 61.0, 61.8, 63.1, 72.6, 76.3, 78.8, 84.9, 85.7, 105.2
MALDI-TOF-MS data.
Compound (If) : calculated m / zn = 0: 834.41, n = 1: 996.46, n = 2: 1158.52, n = 3: 1320.57, n = 4: 1482.62; found m / z [M + Na] ⁺ = 857.4 (n = 0), [M + Na] ⁺ = 1019.4 (n = 1), [M + Na] ⁺ = 1181.5 (n = 2), [M + Na] ⁺ = 1343.7 (n = 3), [M + Na] ⁺ = 1505.6 (n = 4);
Compound (Ig) : calculated m / zn = 0: 848.43, n = 1: 1010.48, n = 2: 1172.53, n = 3: 1334.58, n = 4: 1496.64; found m / z [M + Na] ⁺ = 871.4 (n = 0), [M + Na] ⁺ = 1033.5 (n = 1), [M + Na] ⁺ = 1195.5 (n = 2), [M + Na] ⁺ = 1357.6 (n = 3), [M + Na] ⁺ = 1519.6 (n = 4)

[Test Example 1]
The block methylated cellooligosaccharide library (compounds (If) and (Ig)) obtained in Example 1 was examined for solubility in various solvents.

This library was found to dissolve in any amount in water or methanol. In order to examine the solubility of this library in chloroform (CDCl ₃ ), the library was fractionated according to the method shown in FIG. Was fractionated at a weight ratio of 44:56.

  Table 2 shows the results of analyzing this fractionated library (parts A and B) by MALDI-TOF-MS. In Table 2, the degree of substitution (DS) is a scale representing the average value of substituents introduced into the hydroxyl group of the cellooligosaccharide derivative. Divide by the number of all hydroxyl groups of the sugar derivative and multiply this by 3.

  The content ratio of each cellooligosaccharide derivative contained in the library (compounds (If) and (Ig)) was estimated from the height of the peak detected on the MALDI-TOF-MS spectrum. All of the cellooligosaccharides were water-soluble, but the tetramer and the pentamer were easily dissolved in chloroform, and the oligosaccharide and the hexamer or more were easily dissolved in water. In addition, the hexamer of (21) whose reducing end is a hydroxyl group was particularly highly water-soluble.

  As described above, this synthesis method combining glycosylation and polymerization is effective for model synthesis of block copolymers, and microanalysis using MALDI-TOF-MS is effective for screening the solubility of oligosaccharide derivatives. It was. The block methylated cellooligosaccharide library was amphipathic, but its solubility was confirmed to change depending on the degree of polymerization (DP) and the degree of substitution (DS). .

[Test Example 2]
The surface tension of the aqueous solution of Part A of Test Example 1 was measured using a fully automatic surface tension meter CBVP-Z (manufactured by Kyowa Interface Science Co., Ltd.). The measurement was performed by a Wilhelmi (plate) method under a room temperature condition of 28 ° C.

  While the surface tension of distilled water was about 71 mN / m, the surface tension of the aqueous solution (Part A) containing the cellooligosaccharide derivative of the present invention was about 45 mN / m, confirming the surface activity. FIG. 2 shows the relationship between the compound concentration of the aqueous solution and the surface tension.

  Part A solid sample was dissolved in distilled water, and a concentration solution (mg / mL) was plotted on the horizontal axis and surface tension (nN / m) was plotted on the vertical axis.

From FIG. 2, it was confirmed that the critical micelle concentration (CMC) was about 1 mg / mL.

Example 2 (R ² = Et)
In Example 1 (5), the cellooligosaccharide derivative (R ² = Et) of the present invention was used in the same manner as in Examples 1 (1) to (15) except that ethyl iodide was used instead of methyl iodide. Manufactured. Such a cellooligosaccharide derivative also exhibits an amphipathic property as in Test Example 1 and also has a surface activity as shown in Test Example 2.

[Example 3]
(1) Phenyl 4,6-O-benzylidene-β-D-glucopyranosyl- (1 → 4) -1-thio-β-D-glucopyranoside (49)
To a suspension of phenylthio-β-cellobiloside (15) (5.99 g, 13.7 mM) and benzaldehyde dimethylacetal (3.09 mL, 1.5 eq) in N, N'-dimethylformamide (30 mL), toluenesulfonic acid (0.2 g ) Was added. The reaction was maintained at 45 ° C under 15 mmHg for 3.5 h. Toluenesulfonic acid (1.78 g) was added to the reaction mixture, and the solution was maintained at 50 ° C. under 15 mmHg for 30 minutes. Benzaldehyde dimethylacetal (2.50 mL) was then added and the solution was maintained at 50 ° C. under 15 mmHg for 1 hour. The solution was neutralized with Amberlite IR-400 (OH ⁻ ) and the mixture was filtered to give the crude product. This was purified on a silica gel column (eluent: 10% methanol / CH ₂ Cl ₂ ) to give the title compound (7.1 g, about 100% yield) as a colorless oil.

The title compound (49) was identified by acetylation and conversion to the following compound.
Phenyl 2,3-di-O-acetyl-4,6-O-benzylidene-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-acetyl-1-thio-β-D -Glucopyranoside [Ac form of (49)]
¹ H-NMR (CDCl ₃ ): δ 1.9-2.2 (15H, CH ₃ CO), 3.46 (m, 1H, C _{5 '} -H), 3.61 (m, 1H, C ₅ -H), 3.67 (t, 1H, J _{4 ', 5'} = 9.6, C _{4 '} -H), 3.71 (1H, C ₄ -H), 3.8-3.7 (1H, C _6' -H), 4.08 (dd, 1H, J _{5, 6} = 5.1, J _{6, 6} = 12.0, C ₆ -H), 4.34 (dd, 1H, J _{5 ', 6'} = 4.5, J _{6 ', 6'} = 9.9, C _{6 '} -H), 4.56 ( d, 1H, J _{6, 6} = 18.3, C ₆ -H), 4.58 (d, 1H, J _{1 ', 2'} = 7.5, C _{1 '} -H), 4.66 (d, 1H, J _{1, 2} = 10.2, C ₁ -H), 4.88 (t, 1H, J _{2, 3} = 9.6, C ₂ -H), 4.91 (t, 1H, J _{2 ', 3'} = 9.3, C _{2 '} -H), 5.19 (t, 1H, J _{3, 4} = 9.0, C ₃ -H), 5.25 (t, 1H, J _{3 ', 4'} = 9.6, C _{3 '} -H), 5.47 (s, 1H, -CHPh)
¹³ C-NMR (CDCl ₃ ): δ 20.6, 20.7, 20.7, 20.8, 20.9 (CH ₃ CO), 61.9, 66.3, 68.4, 70.1, 71.9, 72.5, 74.1, 76.5, 76.8, 77.9, 85.2 (C-1 ), 101.5 (C-1 '), 126.0, 128.2, 128.3,128.8, 129.2, 131.4, 133.2, 136.5 (aromatic C), 169.3, 169.5, 169.5, 170.1, 170.2 (CH ₃ CO)
(2) Phenyl 2,3-di-O-benzyl-4,6-O-benzylidene-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-1-thio- β-D-Glucopyranoside (50)
To a tetrahydrofuran (10 mL) solution of the compound (49) obtained in (1) above, 60% NaH (920 mg, 23.0 mM) containing mineral oil and benzyl bromide (2.74 mL, 23.0 mM) were added. The reaction mixture was stirred at room temperature for 11 h and at 50 ° C. for 1 day. Methanol was added thereto at 0 ° C., and the reaction mixture was diluted with ethyl acetate, washed with water and brine, dried over Na ₂ SO ₄ and concentrated to dryness to obtain crystals. The crude crystals were washed with n-hexane to give the title compound (2.54 g, yield 68%) as colorless crystals.

¹ H-NMR (CDCl ₃ ): δ 3.37 (t, 1H, J _{2 ', 3'} = 8.1, C _{2 '} -H), 3.74 (dd, 1H, J _{5, 6} = 1.2, J _{6, 6} = 7.5, C ₆ -H), 3.88 (dd, 1H, J _{5, 6} = 4.5, J _{6, 6} = 7.5, C ₆ -H), 4.21 (dd, 1H, J _{5 ', 6'} = 5.4, J _{6 ', 6'} = 9.9, C _{6 '} -H), (4.57 (d, 1H, J _{1', 2 '} = 7.8, C _1' -H), 4.62 (d, 1H, J _{1, 2} = 9.6 , C ₁ -H), 5.48 (s, 1H, -CHPh)
¹³ C-NMR (CDCl ₃ ): δ 65.8, 68.0 (C ₆ ), 68.7, 73.1, 75.0, 75.5, 75.5, 75.6, 76.6, 79.2, 80.0, 81.2, 81.7, 82.5, 84.9, 87.3 (C-1) , 101.1 (-CHPh), 102.8 (C-1 '), 126.0, 127.6-138.8 (aromatic C)
(3) Phenyl 2,3,6-tri-O-benzyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-1-thio-β-D-glucopyranoside ( 51)
Powder molecular sieve 4A (2.0 g) and sodium cyanoborohydride (0.53 g, 8 mM) were added to a solution of the compound (50) obtained in (2) above (2.00 g, 2.06 mM) in acetonitrile (10 ml). . Trimethylchlorosilane (2.0 ml, 15.8 mM) was slowly added dropwise thereto over 2 hours. The reaction mixture was maintained at room temperature for 3 hours, filtered through Celite 535, and the residue was washed with ethyl acetate. The filtrate was worked up in the usual manner to give a crude product of yellow syrup. This was washed with a silica gel column (Wacogel C-200) eluting with methylene chloride to obtain the title compound (1.28 g, yield 64%) as colorless crystals.

¹ H-NMR (CDCl ₃ ): δ 3.2-3.6 (9H), 3.84 (dd, 1H, J = 3.9, 10.8, H6 or H6 '), 3.74 (dd, 1H, H6 or H6'), 4.00 (t , 1H, J = 9.0, C ₄ -H), 4.50 (d, 1H, J = 7.2, H1 '), 4.63 (d, 1H, J = 9.9, H1), 4.1-5.1 (12H, CH ₂ Ph) , 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 68.1, 71.0, 73.0, 73.4, 73.5, 74.9, 75.3, 75.4, 75.4, 76.3, 76.3, 79.2, 80.0, 82.0, 84.3, 84.8, 87.3 (C-1), 102.4 (C-1 '), 127.3-138.9 (aromatic C)
(4) Phenyl (4-O-acetyl-2,3,6-tri-O-benzyl-β-D-glucopyranosyl)-(1 → 4) -2,3,6-tri-O-benzyl-1- Thio-D-glucopyranoside (52)
To a solution of the compound (51) obtained in (3) above in 2 mL of pyridine was added 2 mL of acetic anhydride at room temperature. The reaction mixture was stirred overnight and concentrated to dryness. The remaining pyridine and acetic anhydride were removed by azeotropy with toluene and ethanol. The crude product was washed with n-hexane to obtain the crystalline title compound (1.17 g, yield 90%).

¹ H-NMR (CDCl ₃ ): δ 1.81 (s, 3H, COCH ₃ ), 3.26-3.55 (7H), 3.61 (t, 1H, J = 8.7), 3.85 (dd, 1H, J = 3.9, 11.1, H6 or H6 '), 3.75 (dd, 1H, H6 or H6'), 4.22 (t, 1H, C ₄ -H), 4.24 (d, 1H, J = 11.4, CH ₂ Ph), 4.37 (d, 1H , J = 11.7, CH ₂ Ph), 4.4-4.9 (9H, CH ₂ Ph), 4.47 (d, 1H, J = 12.3, CH ₂ Ph), 4.52 (d, 1H, J = 7.5, H1 '), 4.63 (d, 1H, J = 9.6, H1), 4.97 (t, 1H, J _{3 ', 4'} = 9.6, J _{4 ', 5'} = 9.9, C _{4 '} -H), 7.2-7.6 (aromatic H )
¹³ C-NMR (CDCl ₃ ): δ 68.1, 71.0, 73.0, 73.1, 73.4, 74.9, 75.3, 75.4, 75.4, 76.3, 79.2, 80.0, 82.0, 84.3, 74.8, 87.3 (C ₁ ), 102.4 (C _{1 '} ), 127.5-128.4 (aromatic C), 128.8, 132.0, 133.6, 137.7, 138.1, 138,2, 138.3, 138.6, 138.9
(5) 4-O-acetyl-2,3,6-tri-O-benzyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-D-glucopyranosyl fluoride ( 53)
To a solution of compound (52) (100 mg, 0.0984 mM) obtained in (4) above in anhydrous methylene chloride (2 mL), DAST (39 μL, 0.295 mM) and NBS (24.5 mg, 0.138 mM) were added at −45 ° C. Added in. The temperature of the reaction mixture was gradually raised to 5 ° C over 2 hours. The mixture was diluted with ethyl acetate, washed with saturated NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give the crude product. The crude product was purified by a silica gel column (eluent; methylene chloride → ethyl acetate: n-hexane = 1: 4 (v / v)) to quantitatively obtain the title compound.

¹ H-NMR (CDCl ₃ ): δ 1.84 (s, 3H, COCH ₃ ), 4.95 (1H, C _{4 '} -H), 5.21 (dd, J _{1β, 2} = 6.4, J _{1β, F} = 53.6, C _1β -H), 5.47 (dd, J _{1α, 2} = 2.7, J _{1α, F} = 50.7, C _1α -H), 7.1-7.5 (aromatic H)
(6) 4-O-acetyl-2,3,6-tri-O-benzyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-D-glucopyranosyl fluoride ( 54)
To a tetrahydrofuran-methanol solution (1.1 mL, 10: 1 (v / v)) of the compound (53) (5 mg) obtained in (5) above, a methanol solution (5 μL) of 28% sodium methoxide was added. The reaction mixture was stirred at room temperature for 1 h. The mixture was diluted with ethyl acetate, washed with water and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give the title compound quantitatively.
(7) 4-O-acetyl-2,3,6-tri-O-benzyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-D-glucopyranose ( 55)
To a solution of the compound (52) (150 mg, 0.148 mM) obtained in (4) above in acetone / water (9: 1, v / v) (5 mL), N-iodosuccinimide (NIS) (65 mg, 0.289 mM) and a catalytic amount of silver trifluoromethanesulfonate (AgOTf) were added at room temperature and the mixture was stirred for 2 h. The reaction mixture was diluted with ethyl acetate, washed with water and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The crude product was purified by preparative thin layer chromatography to give the title compound (55) (103 mg, 76% yield).
(8) 4-O-acetyl-2,3,6-tri-O-benzyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-α-D-glucopyranosyl Immediate (56)
To a solution of compound (55) (103.1 mg, 0.109 mM) obtained in (7) above in anhydrous methylene chloride (5 mL), trichloroacetonitrile (32.8 μL, 0.327 mM) and DBU (8.2 μL, 0.0546 mM) were added at room temperature. And the reaction mixture was stirred for 4.5 h. The mixture was purified with an alumina column to give the title compound quantitatively.
(9) Acetyl 2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-acetyl-D-glucopyranose ( 47)
Lactose (46) (1 g) and sodium acetate (0.5 g) were suspended in acetic anhydride (6 mL) and stirred at 130 ° C. for 35 minutes. The mixture was poured into ice water and crude crystals were collected by filtration. The crude crystals were recrystallized from methanol to obtain the title compound (0.962 g, yield 51%).

¹ H-NMR (CDCl ₃ ): δ 3.75 (m, 1H, C ₅ -H), 3.85 (t, 1H, J _{3, 4} = 8.8, C ₄ -H), 3.87 (dt, 1H, J _{5 ' , 6 '} = 8.0, C _5' -H), 4.13 (dd, 1H, J _{5, 6} = 4.8, J _{5, 6} = 11.6, C ₆ -H), 4.0-4.2 (2H, C _{6 '} -H ), 4.45 (dd, 1H, C ₆ -H), 4.48 (d, 1H, J _{1 ', 2'} = 7.8, C _{1 '} -H), 4.95 (dd, 1H, J _{3', 4 '} = 3.4 , C _{3 '} -H), 5.04 (dd, J _{1, 2} = 8.3, J _{2, 3} = 9.4, C ₂ -H), 5.10 (dd, 1H, C _2' -H), 5.24 (t, 1H , C ₃ -H), 5.35 (dd, 1H, J _{4 ', 5'} = 0.9, C _{4 '} -H), 5.67 (d, 1H, J _{1, 2} = 8.2, C ₁ -H)
(10) 2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-acetyl-α-D-glucopyranosyl bromide (48)
To a solution of compound (47) (0.7584 g, 1.118 mM) obtained in (9) above in anhydrous methylene chloride (5 mL) was added 30% hydrogen bromide in acetic acid (0.668 mL, 3.355 mM) at 0 ° C. added. The reaction mixture was stirred at 0 ° C. for 2 h and then at room temperature for 7 h. The solution was diluted with ethyl acetate, washed with water, saturated NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give the title compound as a colorless oil.
(11) 1,4-Di-O-acetyl-3-O-benzyl-2,6-O-pivaloyl-D-glucopyranose (40)
The title compound was prepared according to Mokuzai Gakkaishi 40, 302-307., Carbohydrate Research 337 (10): 951-954 (2002).
(12) Phenyl 4-O-acetyl-3-O-benzyl-2,6-O-pivaloyl-1-thio-β-D-glucopyranoside (41)
To a solution of compound (40) (69 mg, 0.132 mM) obtained in (11) above in anhydrous methylene chloride (5 mL) was added thiophenol (12.9 μL, 0.126 mM) and boron trifluoride diethyl ether complex (20.1 μL). 0.159 mM) was added at room temperature. After stirring for 11 h, thiophenol (13 μL) was added at room temperature. After 1.5 h, boron trifluoride diethyl ether complex (60 μL) was added at room temperature. After 16.5 h, the reaction mixture was diluted with ethyl acetate, washed with water, saturated NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give the crude product. The product was purified by PTLC (eluent: EtOAc: n-hexane = 1: 4 (v / v)) to give the title compound as crystals (53.5 mg, 71% yield).

¹ H-NMR (CDCl ₃ ): δ 1.21 (s, 9H, COC (CH ₃ ) ₃ ), 1.26 (s, 9H, COC (CH ₃ ) ₃ ), 3.66 (ddd, 1H, C ₅ -H), 3.78 (t 1H, J _{2, 3} = 9.0, C ₃ -H), 4.07 (dd, 1H, J _{5, 6} = 6.0, J _{6, 6} = 12.3, C ₆ -H), 4.24 (dd, 1H, J _{5, 6} = 2.1, J _{6, 6} = 12.3, C ₆ -H), 4.52 (d, 1H, J-11.1), 7.18-7.58 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.7, 27.1, 27.1, 38.7, 38.8, 62.7, 69.2, 70.8, 74.0, 76.4, 81.8, 86.7 (C-1), 127.5, 127.8, 128.0, 128.4, 128.9, 132.4 , 132.8, 137.6, 169.3, 176.4, 178.2
(13) Phenyl 3-O-benzyl-2,6-O-pivaloyl-1-thio-β-D-glucopyranoside (42)
To a solution of the compound (41) (200 mg, 0.350 mM) obtained in (12) above in methanol (20 mL), 1,8-diazabicyclo [5,4,0] -7-undecene (DBU) (366 μL, 2.45 mM)) was added at 0 ° C. and the reaction mixture was stirred at 0 ° C. for 2 h. The reaction mixture was diluted with ethyl acetate, washed with 4N-HCl, saturated NaHCO ₃ aq, water and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give the crude product. The product was purified by PTLC (eluent: EtOAc: n-hexane = 1: 4 (v / v)) to give the title compound as crystals (112.6 mg, 61% yield).

_{^{1 H-NMR (CDCl 3)}} : δ 1.22 (s, 9H, COC (CH 3) 3), 1.26 (s, 9H, COC (CH 3) 3), 3.44-3.62 (C 3 -H, C 4 - H, C ₅ -H), 4.33 (dd, 1H, J _5,6 = 4.5, J _6,6 = 12.3, C ₆ -H), 4.40 (dd, 1H, J _5,6 = 1.8, J _{6, 6} = 12.0, C ₆ -H), 4.67 (d, 1H, J _1,2 = 10.2, C ₁ -H), 4.68 (d, 1H, J = 11.4, CH ₂ Ph), 4.74 (d, 1H, J = 11.4, CH ₂ Ph), 5.02 (dd 1H, J _{1, 2} = 9.0, J _{2, 3} = 9.0, C ₂ -H), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 27.1, 38.7, 38.8, 63.4, 69.8, 71.0, 74.9, 77.9, 83.9, 86.6, 127.6, 127.9, 128.0, 128.6, 128.8, 132.3, 132.9, 137.8, 176.6, 179.0
(14) 4-O-acetyl-3-O-benzyl-2,6-O-pivaloyl-D-glucopyranosyl fluoride (43)
To a solution of compound (41) (200 mg, 0.350 mM) obtained in (12) above in anhydrous methylene chloride (5 mL), (diethylamino) sulfur trifluoride (DAST) (139 μL, 1.05 mM, 3.0 equivalents) and N-bromosuccinimide (NBS) (87.1 mg, 0.489 mM, 1.4 eq) was added at −40 ° C. The temperature of the reaction mixture was gradually raised to 0 ° C. over 5 h. The mixture was diluted with ethyl acetate, washed with saturated NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness to give the crude product. The product was purified by preparative TLC (eluent: EtOAc: n-hexane = 1: 4 (v / v)) to give the title compound as crystals (132.5 mg, 79% yield).
(15) 4-O-acetyl-3-O-benzyl-2,6-O-pivaloyl-D-glucopyranose (44)
N-iodosuccinimide (NIS) (162.4 mg, 0.722) was added to a solution of compound (41) (295.3 mg, 0.516 mM) obtained in (12) above in acetone / water (9: 1, v / v) (10 mL). mM) and a catalytic amount of silver trifluoromethanesulfonate (AgOTf) were added at room temperature and the mixture was stirred for 2 h. The reaction mixture was diluted with ethyl acetate, washed with water and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The crude product was purified by PTLC to give the title compound (180.9 mg, 73% yield).
(16) 4-O-acetyl-3-O-benzyl-2,6-O-pivaloyl-α-D-glucopyranosyl imidate (45)
To a solution of compound (44) obtained in (15) above (180.9 mg, 0.377 mM) in anhydrous methylene chloride (5 mL), trichloroacetonitrile (113 μL, 1.13 mM)) and DBU (28 μL, 0.188 mM) were added at room temperature. added. The reaction mixture was stirred for 3 h and purified on an alumina column to give the title compound quantitatively.
(17) 4-O-acetyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl -(1 → 4) -2,3,6-Tri-O-benzyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-D-glucopyranosyl fluoride (68) (R ² = Me)
To a solution of compound (21) (11 mg, 0.020 mM) obtained in Example 1 (9) and compound (54) (15 mg, 0.017 mM) obtained above (6) in anhydrous methylene chloride (1 mL) was added MS4A. (100 mg) was added and stirred at room temperature for 1.5 h. To the mixture was added NIS (13.3 mg, 0.059 mM) and catalytic amount of AgOTf (1.5 mg) at −78 ° C. The reaction mixture was gradually warmed to room temperature and stirred for 22 hours. Insoluble material was filtered off and washed with ethyl acetate. The filtrate was diluted with ethyl acetate, washed with distilled water, NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The product was separated by PTLC (eluent: EtOAc: n-hexane = 1: 2 (v / v) twice) to give the title compound (4 mg, 18% yield).

Calculated for C ₇₄ H ₉₁ FO ₂₁ : 1334.6; ESI-MS: [M + Na] ⁺ = 1357
¹ H-NMR (CDCl ₃ ): δ 3.18, 3.32, 3.46, 3.48, 3.52, 3.63 (OCH ₃ ), 5.46 (dd, C1-Hα), 5.47 (dd, C1-Hβ)
(18) Production Method of Compound (69) (R ² = Me) Compound (68) obtained in the above (17) is reacted with Cp ₂ HfCl ₂ / AgClO ₄ in a water-containing solvent (for example, acetone-water) to give −F Was converted to —OH, debenzylated by catalytic reduction with Pd (OH) ₂ -carbon under a stream of hydrogen, and deesterified with sodium methoxide to give the title compound.

[Example 4]
(1) Phenyl 4-O-acetyl-2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D- Glucopyranosyl- (1 → 4) -3-O-benzyl-2,6-di-O-pivaloyl-1-thio-β-D-glucopyranoside (70) (R ² = Me)
Compound (42) (10 mg, 0.019 mM) obtained in Example 3 (13) and compound (22) (17.7 mg, 0.038 mM) obtained in Example 1 (10) in anhydrous benzene (2 mL) To the solution was added active MS4A (200 ml) and stirred at room temperature for 3 h. To the reaction mixture, Cp ₂ HfCl ₂ (15.7 mg, 0.041 mM) and AgClO ₄ (17.2 mg, 0.083 mM) were added at room temperature and stirred for 2.5 h. Insoluble material was filtered off and washed with ethyl acetate. The filtrate was diluted with ethyl acetate, washed with distilled water, NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The product was purified by PTLC (eluent: EtOAc: n-hexane = 1: 2 (v / v), several times) to give the title compound (6.0 mg, 32% yield).

Calculated for C ₄₉ H ₇₂ O ₁₈ S: 980.44; MALDI-TOF MS: [M + Na] ⁺ = 1003.17
¹ H-NMR (CDCl ₃ ): δ 1.17, 1.22 (s, s, 9H, 9H, COC (CH ₃ ) ₃ ), 2.08 (s, 3H, COCH ₃ ), 3.08, 3.34, 3.48, 3.49, 3.55 ( s, 3H, 3H, 3H, 3H, 6H, OCH ₃ ), 2.94-3.02 (t, t, C _{2 '} -H and C _2'' -H), 3.12-3.24 (t, t, C _3'- H and C _{3 ''} -H), 3.3-3.6 (C _{6 ''} -H), 3.36-3.44 (C _{5 ''} -H), 3.6-3.7 (m, 1H, C ₅ -H), 3.71 ( t, 1H, J = 8.4, C ₃ -H), 3.82 (t, 1H, J = 9.0, C ₄ -H), 4.20 (dd, 1H, J = 6.0, J = 12.0, C ₆ -H), 4.30 (d, 1H, J = 7.8, C 1 '-H or C 1''-H), 4.35 (d, 1H, J = 8.1, C 1' -H or C 1 '' -H), 4.58 ( d, 1H, J = 11.7, CH ₂ Ph), 4.66 (d, 1H, J = 10.2, C ₁ -H), 4.71 (dd, 1H, J = 2.1, J = 12.3, C ₆ -H), 4.84 (t, 1H, J = 9.5, C _{4 ''} -H), 5.04 (dd, 1H, J = 9.0, J = 10.2, C ₂ -H), 5.08 (d, 1H, J = 11.4, CH ₂ Ph ), 7.18-7.50 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.9, 27.0, 27.2, 38.7, 38.8, 58.7, 59.5, 60.3, 60.4, 60.6, 63.0, 69.7, 70.9, 72.1, 73.0, 74.3, 75.0, 78.2, 83.0, 83.5, 83.9, 85.0, 86.5, 103.0, 103.5, 126.6, 127.0, 127.8, 128.0, 128.8, 132.4, 133.2, 138.7, 169.8, 177.9
(2) Preparation of Compound (71) (R ² = Me) Compound (70) obtained in (1) above was treated with NIS / AgOTf to convert -SPh to -OH, and Pd ( Debenzylation by catalytic reduction with OH) ₂ -carbon and deesterification with sodium methoxide gave the title compound.

[Example 5]
(1) Phenyl 4-O-acetyl-3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl- β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D-glucopyranoside (59) (R ² = Me)
Compound (20) obtained in Example 1 (8) (12.2 mg, 0.024 mM) and compound (43) (22.7 mg, 0.047 mM) obtained in Example 3 (14) in anhydrous benzene (2 mL) To the solution was added active MS4A (200 mg), and the mixture was stirred at room temperature for 1.5 h. To the reaction mixture, Cp ₂ HfCl ₂ (19.7 mg, 0.052 mM) and AgClO ₄ (21.5 mg, 0.104 mM) were added at room temperature and stirred for 3.5 h. Insoluble material was filtered off and washed with ethyl acetate. The filtrate was diluted with ethyl acetate, washed with distilled water, NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The product was separated by PTLC (eluent: EtOAc: n-hexane = 1: 2 (v / v)) to give the title compound (12.7 mg, 55% yield).

Calculated for C ₄₉ H ₇₂ O ₁₈ S: 980.44; MALDI-TOF MS: [M + Na] ⁺ = 1003.29
¹ H-NMR (CDCl ₃ ): δ 1.19, 1.21 (s, s, 9H, 9H, COC (CH ₃ ) ₃ ), 1.93 (s, 3H, COCH ₃ ), 3.38, 3.42, 3.52, 3.57, 3.59 ( s, 3H, 3H, 3H, 3H, 6H, OCH ₃ ), 2.88 (dd, 1H, J = 7.8, J = 9.0, C _{2 '} -H), 3.07 (dd, 1H, J = 8.7, J = 9.9 , C ₂ -H), 3.17 (t, 1H, J = 9.3, C _{3 '} -H), 3.15-3.22 (C _5' -H), 3.26 (t, 1H, J = 8.4, C ₃ -H) _{, 3.34-3.42 (C 5 -H),} 3.56-3.64 (1H, C 5 '' -H), 3.64-3.76 (1H, C 4 -H), 3.68-3.76 (C 6 -H and C 6 '- H), 3.73-3.78 (1H, C _{3 ''} -H), 3.74-3.80 (1H, C _{4 '} -H), 4.15 (dd, 1H, J = 12.6, J = 3.0, C _6'' -H ), 4.20 (dd, 1H, J = 4.2, J = 12.3, C _{6 ''} -H), 4.27 (d, 1H, J = 7.8, C _{1 '} -H), 4.50 (d, 1H, J = 9.9 , C ₁ -H), 4.53 (d, 1H, J = 12.3, CH ₂ Ph), 4.61 (d, 1H, J = 8.1, C _{1 ''} -H), 4.63 (d, 1H, J = 11.4, _{CH 2 Ph), 5.09 (dd} , 1H, J = 8.1, J = 9.6, C 2 '' -H), 5.19 (t, 1H, J = 9.6, C 4 '' -H), 7.19-7.55 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.7 (COCH ₃ ), 27.1 (COC (CH ₃ ) ₃ ), 27.2 (COC (CH ₃ ) ₃ ), 38.8 (COC (CH ₃ ) ₃ ), 59.1 (C ₆ OCH ₃ or C _{6 '} OCH ₃ ), 59.4 (C ₆ OCH ₃ or C _6' OCH ₃ ), 60.7 (C ₂ OCH ₃ or C _{2 '} OCH ₃ , or C ₃ OCH ₃ or C _3' OCH ₃ ), 60.8 (C ₂ OCH ₃ or C _{2 '} OCH ₃ , or C ₃ OCH ₃ or C _3' OCH ₃ ), 62.1 (C _{6 ''} ), 69.3 (C _{4 ''} ), 70.3 (C ₆ or C _{6 '} ), 70.4 (C ₆ or C _{6 '} ), 72.1 (C _5'' ), 72.5 (C _2'' ), 73.2 (CH ₂ Ph), 74.6 (C _5' ), 75.8 (C _{4 '} ), 77.8 (C ₄ ), 79.0 (C ₅ ), 80.8 (C _{3 ''} ), 82.0 (C ₂ ), 83.4 (C _{2 '} ), 84.7 (C _3' ), 86.8 (C ₃ ), 87.3 (C ₁ ) , 99.9 (C _{1 ''} ), 103.3 (C _{1 '} ), 127.3, 127.5, 127.7, 128.4, 128.8, 131.8, 133.9, 137.6 (aromatic C), 169.2 (COCH ₃ ), 176.5 (COC (CH ₃ ) ₃ ), 178.2 (COC (CH ₃ ) ₃ )
(2) Preparation of compound (60) (R ² = Me) Compound (59) obtained in (1) above was treated with NIS / AgOTf / acetone-H ₂ 0 to convert -SPh to -OH. To give the title compound.
(3) Preparation of Compound (61) (R ² = Me) Compound (59) obtained in (2) above was debenzylated by catalytic reduction with Pd (OH) ₂ -carbon in a hydrogen atmosphere, and sodium methoxide. To give the title compound.

[Example 6]
(1) Phenyl 2,3,4,6-tetra-O-acetyl-β-D-galactopyranosyl- (1 → 4) -2,3,6-tri-O-acetyl-β-D-glucopyranosyl -(1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D -Glucopyranoside (62) (R ² = Me)
Anhydrous methylene chloride (5 mL) of the compound (20) obtained in Example 1 (8) (10 mg, 0.019 mM) and the compound (48) obtained in Example 3 (10) (26.9 mg, 0.039 mM) Active MS4A (200 mg) was added to the solution and stirred at room temperature for 4 h. To the reaction mixture, AgOTf (19.8 mg, 0.077 mM) was added at room temperature and stirred for 3 days. Insoluble material was filtered off and washed with ethyl acetate. The filtrate was diluted with ethyl acetate, washed with distilled water, NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The product was separated by PTLC (eluent: EtOAc: n-hexane = 2: 1 (v / v) to give the title compound (1.8 mg, 6% yield).

¹ H-NMR (CDCl ₃ ): δ 2.92 (dd, 1H, J = 8.1, J = 8.7, C _{2 '} -H), 3.07 (dd, 1H, J = 8.7, J = 9.9, C ₂ -H) , 3.17 (t, 1H, J = 9.0, C _{3 '} -H), 3.18-3.26 (C _5' -H), 3.29 (t, 1H, J = 9.0 C ₃ -H), 3.3-3.44 (m) , 3.37, 3.40, 3.52, 3.53 (3H, 3H, 3H, 3H, OCH ₃ ), 3.59 (6H, OCH ₃ ), 3.5-3.9 (m), 4.07 (dd, J = 7.2, J = 11.7), 4.10 (t, 1H, J = 7.2), 4.17 (dd, 1H, J = 3.9, J = 11.1), 4.28 (d, 1H, J = 7.5), 4.36-4.56 (3H), 4.71 (d, 1H, J = 8.1, 4.88 (t, 1H, J = 7.8), 4.95 (dd, 1H, J = 3.3, J = 10.8), 5.10 (dd, 1H, J = 7.5, J = 9.9), 5.17 (t, 1H, J = 9.0), 5.35 (dd, 1H, J _{3 ', 4'} = 3.6, C _{4 '''} -H)
(2) Preparation of Compound (63) (R ² = Me) Compound (62) obtained in (1) above was treated with NIS / AgOTf / acetone-H ₂ 0 to convert -SPh to -OH. To give the title compound.
(3) Preparation of compound (64) (R ² = Me) Compound (63) obtained in (2) above was deesterified with sodium methoxide to give the title compound.

[Example 7]
(1) Phenyl 4-O-acetyl-2,3,6-tri-O-benzyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-β-D- Glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β- D-Glucopyranoside (65) (R ² = Me)
Compound (56) (38.9 mg, 0.036 mM) obtained in Example 3 (8) and compound (20) (10 mg, 0.019 mM) obtained in Example 1 (8) in anhydrous methylene chloride (1.0 mL) To the solution was added boron trifluoride diethyl ether complex (0.24 μL, 0.0019 mM) at −78 ° C. The reaction temperature was gradually raised to −50 ° C. and stirred for 2 h. Boron trifluoride diethyl ether complex (1.0 μL) was added again at −50 ° C. and stirred for 1 day. The mixture was diluted with ethyl acetate, washed with saturated NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The product was separated by PTLC (eluent: EtOAc: n-hexane = 1: 2 (v / v)) to give the title compound (22.4 mg, 81% yield).

¹ H-NMR (CDCl ₃ ): δ 1.8 (3H, COCH ₃ ), 2.95 (dd, 1H, J = 7.8, J = 9.0, C _{2 '} -H), 3.09 (dd, 1H, J = 8.7, J = 9.6, C ₂ -H), 3.18 (t, 1H, J = 8.7, C _{3 '} -H), 3.28, 3.40, 3.55, 3.56, 3.59, 3.60 (3H, 3H, 3H, 3H, 3H, 3H, OCH ₃ ), 4.04 (t, 1H, J = 9.6, C _{4 ''} -H), 4.31 (d, 1H, J = 7.8, C _{1 '} -H), 4.43 (d, 1H, J = 7.5, C _{1 ''} -H), 4.51 (d, 1H, J = 10.2, C ₁ -H), 4.56 (d, 1H, J = 7.2, C _{1 '''} -H), 4.96 (t, 1H, J = 8.7, C _{4 '''} -H), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.8 (COCH ₃ ), 59.0 (OCH ₃ ), 59.2 (OCH ₃ ), 60.7 (OCH ₃ ), 60.7 (OCH ₃ ), 60.8 (OCH ₃ ), 67.7, 69.9, 70.4, 71.5 (C _{4 '''} ), 73.3, 73.5, 74.7, 75.1, 75.2, 76.4 (C _4'' ), 76.5-77.0 (C _4' ), 77.9, 78.9, 82.0 (C ₂ ), 82.5, 83.2, 83.4 (C _{2 '} ), 85.0 (C _3' ), 86.7 (C ₃ ), 87.2 (C ₁ ), 102.2 (C _{1 '''} ), 102.8 (C _1'' ), 103.4 (C _1' ), 127.3, 127.4, 127.5, 127.5, 127.7, 127.7, 127.9, 127.9, 128.0, 128.2, 128.2, 128.3, 128.8, 131.7, 133.8, 136.5 ,, 138.1, 138.3, 138.3, 139.0, 169.9 (COCH ₃ )
(2) Preparation of Compound (66) (R ² = Me) Compound (65) obtained in (1) above was treated with NIS / AgOTf / acetone-H ₂ 0 to convert -SPh to -OH. To give the title compound.
(3) Preparation of Compound (67) (R ² = Me) Compound (66) obtained in (2) above was debenzylated by catalytic reduction with Pd (OH) ₂ -carbon in a hydrogen atmosphere, and sodium methoxide. To give the title compound.

[Example 8]
(1) Preparation of Compound (37) Compound (37) was obtained from Compound (16) obtained in Example 1 (4) according to the method described in Reaction Scheme 8 in the text.
(2) Preparation of Compound (57) (R ² = Me) Compound (37) (11.3 mg, 0.012 mM) obtained in (1) above and Compound (20) (1) obtained in Example 1 (8) ( Active MS4A (200 mg) was added to a solution of 5.6 mg, 0.011 mM) in anhydrous benzene (2 mL), and the mixture was stirred at room temperature for 1.5 h. To the reaction mixture, Cp ₂ HfCl ₂ (5.0 mg, 0.013 mM) and AgClO ₄ (5.4 mg, 0.026 mM) were added at room temperature and stirred for 3 days. The insoluble material was filtered off and washed with ethyl acetate. The filtrate was diluted with ethyl acetate, washed with distilled water, NaHCO ₃ aq and brine, dried over Na ₂ SO ₄ and concentrated to dryness. The product was isolated by PTLC (eluent: EtOAc: n-hexane = 1: 2 (v / v) to give the title compound (1.6 mg, 10% yield).
(3) Preparation of Compound (58) (R ² = Me) Compound (57) obtained in (2) above was deesterified with sodium methoxide to give the title compound.

[Example 9]
(1) Phenyl 4-O-acetyl-2,3,6-tri-O-benzyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-benzyl-β-D- Glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-Tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D- Glucopyranoside (86) (R ² = Me)
In a glass reaction tube connected to a vacuum line, the compound (56) obtained in Example 3 (8) (40 mg, 0.0375 mmol, 1.4 equiv.) And the compound (26) obtained in Example 1 (8) were used. (25.5 mg, 0.0276 mmol, 1.0 equiv.) Was added and dried overnight under high vacuum. Anhydrous methylene chloride (1 mL) was transferred to a glass reaction tube in a vacuum line, and while the reaction tube part was cooled with liquid nitrogen, the connecting glass part of the glass reaction tube and the vacuum line was burned out using a gas burner. The reaction tube was transferred to a -78 ° C constant temperature bath and stirred. Boron trifluoride ether complex (0.24 μL, 1.87 × 10 ⁻³ mmol, 0.05 mol%) was added and reacted at −78 ° C. for 27.5 hours. Thereafter, the reaction solution was extracted with ethyl acetate, washed with saturated sodium bicarbonate, distilled water and saturated brine, dried over sodium sulfate, and concentrated. The reaction mixture was separated and purified by PTLC (eluent: EtOAc / n-hexane = 1: 1, v / v, 2 times) to give the title compound (14.7 mg, 5.95 × 10 ⁻³ mmol, yield 29.1%). Obtained.

¹ H-NMR (CDCl ₃ ): δ 1.8 (3H, COCH ₃ ), 2.91-3.00 (3H, t, t, t), 3.09 (dd, 1H, J = 9, 9.6, H2), 3.28, 3.38, 3.39, 3.41, 3.54, 3.56, 3.57, 3.59, 3.60, 3.62 (OCH ₃ ), 3.88 (dd, 1H, H6`` ''), 4.04 (t, 1H, J = 9.3, H4 ''''), 4.22 (d, 1H, J = 11.7, CH <sub>2</sub> Ph), 4.32 (d, 1H, J = 7.8, H1 'or, H1''orH1'''), 4.32-4.36 (2H, H1 'or, H1 '' or H1 '''), (4.37 (d, 1H, J = 11.7, CH <sub>2</sub> Ph), 4.43 (d, 1H, J = 7.5, H1``''), 4.51 (d, 1H, J = 9.9, H1), 4.56 (d, 1H, J = 7.2, H1 '''''), 4.61 (d, 1H, J = 11.4, CH ₂ Ph), 4.66 (d, 1H, J = 11.4, CH ₂ Ph), 4.68-4.84 (7H, CH ₂ Ph), 4.96 (t, 1H, J = 9.0, H4 '''''), 5.01 (d, 1H, J = 11.1, CH ₂ Ph), 7.2-7.6 (aromatic H)
¹³ C-NMR (CDCl ₃ ): δ 20.8 (COCH ₃ ), 59.0, 59.1, 59.2, 59.2, 60.3, 60.4, 60.6, 60.6, 60.7, 60.7, 60.8 (OCH ₃ ), 67.8 (C6`` '') , 70.0, 70.2, 70.5, 71.6 (C4 '''''), 73.3, 73.5, 74.9, 75.1, 75.2, 75.3, 76.4 (C4''''), 76.8, 77.2, 77.6, 77.8, 79.0, 82.1, 82.5, 83.3, 83.5, 83.6, 85.1, 85.1, 85.1, 86.8, 87.3 (C1), 102.2 (C1 '''''), 102.8 (C1''''), 103.1 (C1' or C1 '' or C1 '''), 103.2 (C1' or C1 '' or C1 '''), 103.3 (C1' or C1 '' or C1 '''), 127.3, 127.4, 127.5, 127.7, 127.7, 127.9, 128.0, 128.0 , 128.2, 128.2, 128.3, 128.8, 131.7, 133.9, 138.2, 138.3, 138.3, 138.4, 138.4, 139.1 (aromatic C), 169.9 (COCH ₃ )
MALDI-TOF: calculated m / z 1832.82; found m / z: found [M + Na] ⁺ = 1855.795
(2) Preparation of Compound (87) (R ² = Me) Compound (86) obtained in (1) above was treated with NIS / AgOTf / acetone-H ₂ 0 to convert -SPh to -OH. To give the title compound.
(3) Preparation of Compound (88) (R ² = Me) Compound (87) obtained in (2) above was debenzylated by catalytic reduction with Pd (OH) ₂ -carbon in a hydrogen atmosphere to obtain sodium methoxide. To give the title compound.

[Example 10]
(1) Phenyl 4-O-acetyl-3-O-benzyl-2,6-di-O-pivaloyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl- β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β-D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-β- D-glucopyranosyl- (1 → 4) -2,3,6-tri-O-methyl-1-thio-β-D-glucopyranoside (89) (R ² = Me)
The compound (43) (10.8 mg, 0.0224 mmol, 2.1 equiv.) Obtained in Example 3 (14) and the compound (26) obtained in Example 1 (8) were placed in a glass reaction tube connected to a vacuum line. 10 mg, 0.0107 mmol, 1.0 equiv.) And Molecular Sieves 4A (500 mg) were added and dried overnight under high vacuum. Anhydrous benzene (5 mL) was transferred to a glass reaction tube in a vacuum line. Cp ₂ HfCl ₂ (9.1 mg, 0.0237 mmol, 1,1 equiv.) And AgClO ₄ (9.9 mg, 0.0475 mmol, 2.2 equiv.) Were transferred to a reaction vessel in a vacuum line, and the reaction was started at room temperature. After 19 hours, filtered through celite and washed with EtOAc. The filtrate was diluted with ethyl acetate, washed with saturated sodium bicarbonate, distilled water and saturated brine, dried over sodium sulfate, and concentrated. The reaction mixture was separated and purified by PTLC (EtOAc / n-Hexane = 2: 1, v / v,) to obtain the title compound (4.6 mg, 3.31′10 ⁻³ mmol, yield 30.1%).

¹ H-NMR (CDCl ₃ ): δ 1.19, 1.22 (9H, 9H, C2-O-COC (CH ₃ ) ₃ , C6-O-COC (CH ₃ ) ₃ , respectively), 1.93 (3H, COCH ₃ ) , 2.88 (dd, 1H, J = 7.8, 9.3, H2 '''), 2.9-3.0 (dd, dd, 2H, H2' and H2 ''), 3.08 (dd, 1H, J = 8.7, 9.9, H2 ), 3.13-3.24 (3H, H3 'and H3''andH3'''), 3.26-3.42 (H5, H5 ', H5'', and H5'''), 3.28 (t, J = 8.7, H3 ), 3.38, 3.38, 3.39, 3.42, 3.52, 3.54, 3.56, 3.57, 3.58, 3.60, 3.61 (OCH ₃ ), 3.58-3.66 (H5`` ''), 3.66-3.80 (H4, H4 ', and H4 ''), 3.75-3.84 (H4 '''), 3.65-3.84 (H6, H6', H6 '', and H6 '''), 4.15 (dd, 1H, J = 3.0, H6'''') , 4.21 (dd, 1H, J = 4.2, 12.6, H6`` ''), 4.29 (d, 1H, J = 7.2, H1 '''), 4.31 (d, 1H, J = 7.6), 4.34 (d , 1H, J = 7.8), 4.51 (d, 1H, J = 9.9, H1), 4.53 (d, 1H, J = 11.1, CH ₂ Ph), 4.61 (d, 1H, J = 7.8, H1 ''''), 4.63 (d, 1H, J = 11.1, CH ₂ Ph), 5.09 (dd, 1H, J = 7.8, 9.3, H2``''), 5.19 (t, 1H, J = 9.3, H4'''')
<sup>13</sup> C-NMR (CDCl <sub>3</sub> ): δ 20.7 (COCH <sub>3</sub> ), 27.1, 27.2 (COC (CH <sub>3</sub> ) <sub>3</sub> ), 38.8 (COC (CH <sub>3</sub> ) <sub>3</sub> ), 59.2, 59.4, 60.4, 60.6, 60.7, 60.8 ( OCH <sub>3</sub> ), 62.1 (C6`` ''), 69.2 (C4 ''''), 70.2 (C6 and / or C6 'and / or C6''and / or C6'''), 70.5 (C6 and / or C6 'and / or C6''and / or C6'''), 72.1 (C5`` ''), 72.5 (C2 ''''), 73.2 (CH <sub>2</sub> Ph), 74.5 (C5 'and / or C5 '' or C5 '''), 74.7 (C5' or C5 '' or C5 '''), 74.8 (C5' or C5 '' or C5 '''), 75.9 (C4'''), 77.8 ( C4 and C4 'and C4``), 79.0 (C5), 80.8 (C3''''), 82.0 (C2), 83.4 (C2' and / or C2 '' and / or C2 '''), 83.5 ( C2 'and / or C2''and / or C2'''), 84.8 (C3 'and / or C3''and / or C3'''), 85.0 (C3 'and / or C3''and / or C3 '''), 86.7 (C3), 87.3 (C1), 99.9 (C1''''), 103.1 (C1' and / or C1 '' and / or C1 '''), 103.3 (C1' and / or C1 '' and / or C1 '''), 127.3, 127.5, 127.7, 128.4, 137.6 (aromatic C), 169.2 (COCH ₃ ), 176.5, 178.2 (COC (CH ₃ ) ₃ )
MALDI-TOF: calculated m / z 1388.64 found m / z: found [M + Na] ⁺ = 1411.319
(2) Preparation of Compound (90) (R ² = Me) Compound (89) obtained in (1) above was treated with NIS / AgOTf / acetone-H ₂ 0 to convert -SPh to -OH. To give the title compound.
(3) Preparation of Compound (91) (R ² = Me) Compound (90) obtained in (2) above was debenzylated by catalytic reduction with Pd (OH) ₂ -carbon in a hydrogen atmosphere, and sodium methoxide. To give the title compound.

[Test Example 3] (Light scattering method)
In order to know the association state of the cellooligosaccharide derivative in an aqueous solution, the shape of the aggregate (nanoparticle) composed of the cellooligosaccharide derivative was observed using a light scattering method.
(1) Measurement conditions for the radius of aggregates (nanoparticles) About the aqueous solution of Part A obtained in Test Example 1, using a light scattering photometer (SLS-5000HM, manufactured by Otsuka Electronics Co., Ltd.), the following conditions Static light scattering was measured. Dynamic light scattering (DLS) was measured using a light scattering photometer (SLS-5000HM, manufactured by Otsuka Electronics Co., Ltd.) equipped with ALV-5000 / E correlator (ALV Co.).

Wavelength: λ ₀ = 632.8 nm (helium-neon laser)
Scattering angle: measured every 15 ° from 30 ° to 150 °.

Average temperature (Avg. Temp.): 25.08 ° C
Refractive index increment: dn / dc = 0.1340 mL / g (methyl cellulose): Polymer Handbook 4th edition / editors, J. Brandrup, EH Immergut, and EA Grulke; associate editors, A. Abe, DR Bloch; New York: Wiley, c1999 Referred to.

The aqueous solution of Part A was filtered several times with PTFE membrane filter paper (manufactured by Toyo Filter Paper Co., Ltd.) having a pore size of 0.2 μm, and then measurement was performed.
(2) The analysis result of the radius of the aggregate (nanoparticle) and the association number static light scattering is shown below. These analyzes were carried out in accordance with “New Polymer Experimental Science 1 Basic Molecular Analysis of Polymer Experiments: edited by the Society of Polymer Science, Kyoritsu Shuppan October 25, 1994”.

Cellooligosaccharide of Part A: Weight average molecular weight Mw = 1003 (estimated from peak intensity of MALDI-TOF MS spectrum)
The mean square inertial radius of nanoparticles on the high and low concentrations (see Figure 2) is as follows:
<High concentration side (0.349-0.5048 mg / mL)>
Nanoparticle weight average molecular weight (Mw) = 6.947 × 10 ⁵ ; Aggregation Number = 693 (divided by Mw = 1003)
Second virial coefficient (A ₂ ) = 3.119 × 10 ^-3 cm ³ g ^-2 mol
Mean square inertial radius <R _g ² > ^1/2 = 111.8 nm
<Low concentration side (0.200284-0.30105 mg / mL)>
Nanoparticle weight average molecular weight (Mw) = 2.975 × 10 ⁵ ; Aggregation Number = 297 (divided by Mw = 1003)
Second virial coefficient (A ₂ ) = 9.912 × 10 ^-4 cm ³ g ^-2 mol
Mean square inertial radius <R _g ² > ^1/2 = 52.17 nm
From these results, it was found that the cellooligosaccharide derivative having surface active ability changes the mean square inertial turning radius depending on the concentration in the aqueous solution. It was found that the higher the concentration, the larger the size of the nanoparticles in the aqueous solution. Further, it was found that the weight average molecular weight of the nanoparticles calculated by the light scattering method was extremely large compared to the weight average molecular weight Mw = 1003 of the synthesized cellooligosaccharide derivative Part A. Therefore, it was concluded that the nanoparticles were aggregates in which many cellooligosaccharide derivatives were associated as in the above calculation results. In general, for the second virial coefficient, among solvents that dissolve the polymer, a solvent having a high affinity with the polymer (good solvent) is a second virial coefficient A of 10 ⁻⁴ mol cm ³ / g ² or more. ₂ , low solvent (poor solvent) gives a lower value. Therefore, it was found that for the synthesized cellooligosaccharide derivative Part A, water as a solvent is a good solvent.

  On the other hand, when a Zimm plot was performed from measurement data of light scattering up to 0.200284-0.30105 mg / mL, the calculated results are shown below.

Static light scattering: Mean square inertial radius <R _g ² > ^1/2 64.4 nm
Dynamic light scattering: hydrodynamic radius R _h 45.4 nm
R _g / R _h = 1.42
From this value, it can be determined that the particles are rod-like rather than spherical. (The closer the value of R _g / R _h is to 1, the closer it is to a true sphere.)
Summarizing the above results, it was shown by calculation that the molecular aggregate (aggregate) gradually increases in size when the surfactant composed of cellooligosaccharide becomes high in concentration.

Moreover, it was found that the cellooligosaccharide derivative of the present invention has a relatively large number of associations as a nonionic surfactant. It was also shown that the particles are rod-like structures rather than spheres. It was shown that the association number of the surfactant increases as the concentration of the compound increases. It has not been known so far that such an oligosaccharide having a low molecular weight as a single molecule forms a huge aggregate compared with the molecular size. Such a phenomenon could be analyzed by a physicochemical method called light scattering.

[Test Example 4] (TEM observation)
After 2.5 μL of 1 mg / mL cellooligosaccharide aqueous solution (part A aqueous solution of Test Example 1) was mixed with 2.5 μL of a saturated aqueous solution of uranyl acetate (ca. 2.0 wt%) on a copper grid coated with a polymer thin film, It was dried under reduced pressure and washed 3 times with distilled water.

The obtained particles were observed with a transmission electron microscope (TEM, JEOL JEM 1220) at an acceleration voltage of 100 KeV. The magnification (Magnification) was 25,000 times. The concentration of the cellooligosaccharide aqueous solution at the time of measurement was 1.0 mg / mL. Negative staining with uranyl acetate. As shown in FIG. 3, rugby ball-type slightly elongated particles were observed, that is, it was actually confirmed that the cellooligosaccharide derivative self-assembled in an aqueous solution to form a nanoparticle structure. The particle size was consistent with the above light scattering data.

[Test Example 5] (Atomic force microscope observation)
A 1 mg / mL cellooligosaccharide aqueous solution (part A aqueous solution of Test Example 1) was sonicated for 10 s and dropped onto mica. Subsequently, after natural drying, it observed with the atomic force microscope (AFM). As the atomic force microscope, Olympus NV2000 (manufactured by Olympus Corporation) was used. Measurement was performed using a cantilever (Olympus AC240TS-C1) in a non-contact mode.

  4 (a) and 4 (b) are observations of nanoparticles at different locations on the mica. 4 (a) and 4 (b), it can be said that the same particle structure was observed no matter which place was observed.

  FIG. 4B and FIG. 5 are the observation results of the nanoparticles at the same place, and are shown by different display methods to show the height (particle diameter) of the nanoparticles. As a result, a particulate structure having a height of about 30 to 300 nm was observed. Since there was no change in the height direction of the particles even after vacuum drying, it was found that the particulate structure was a nanoparticle filled inside.

  In FIG. 5, the shape of the cross section along the oblique line drawn from the upper right to the lower left in the left square portion is shown in the right rectangular window. Each mountain represents a nanoparticle composed of one oligosaccharide. Since AFM has the feature that the length in the height direction can be measured accurately, the heights of the nanoparticles are relatively uniform, and the height data is consistent with the data obtained by the light scattering method in Test Example 3. I understood it.

  TEM images are measured under high vacuum, and the particle morphology may be deformed, but AFM observation is observed in the atmosphere under atmospheric pressure, and the possibility that the particle morphology will change is low. These data are consistent with the actual light scattering experimental data in solution, which suggests that the structure of nanoparticles composed of oligosaccharides is stable even when dried in solution.

Claims (12)

A surfactant comprising a cellooligosaccharide derivative in which a hydrophobic moiety comprising an alkylated glucose or cellooligosaccharide and a hydrophilic moiety comprising a monosaccharide or oligosaccharide are bound in a block manner ,
General formula:
HBA AOR ¹
H-A-B-A-OR ¹
R ¹ -A-B-OH or
R ¹ -B-A-B-OH
(In the formula, A is a general formula:
(In the formula, R ² represents C _1-4 alkyl, and m represents an integer of 1-8.)
B is composed of a hydrophilic part obtained by removing an anomeric hydroxyl group from a pyranose type monosaccharide and removing -H from one alcoholic hydroxyl group, or a pyranose type monosaccharide. 2 or more selected from the group except for the anomeric hydroxyl group of the reducing terminal sugar residue from the oligosaccharide having a 1,4-glycoside bond and -H from the alcoholic hydroxyl group at the 4-position of the non-reducing terminal sugar residue The resulting hydrophilic part, when two or more A are contained, A may be the same or different from each other, and when two or more B is contained, B may be the same or different from each other, and R ¹ is H or C _1-4 alkyl is shown. However, the sugar chain is arranged so that the reducing end is on the right side of the page. )
A surfactant comprising a cellooligosaccharide derivative represented by: General formula:
  H-B-A-OR ¹   Or
  R ¹ -A-B-OH
(Wherein R ¹ , A and B are the same as above)
The surfactant according to claim 1, comprising the cellooligosaccharide derivative shown. R ¹ Is a monosaccharide selected from the group consisting of glucose, galactose, mannose, xylose, ribose, N-acetylglucosamine, glucosamine, arabinose, rhamnose, fucose, N-acetylgalactosamine, and galactosamine The surfactant according to claim 1 or 2, comprising a cellooligosaccharide derivative. The surfactant according to claim 3, comprising a cellooligosaccharide derivative in which the pyranose type monosaccharide is glucose. Formula (I):
(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
The surfactant according to claim 1, comprising a cellooligosaccharide derivative represented by the formula: General formula (II):
(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m is an integer of 1-8, and n is an integer of 1-8.)
The surfactant according to claim 1, comprising a cellooligosaccharide derivative represented by the formula: General formula (III):
(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m ₁ and m ₂ are the same or different and are an integer of 1-8, n ₀ is an integer of 1-8. .)
The surfactant according to claim 1, comprising a cellooligosaccharide derivative represented by the formula: Formula (IV):
(In the formula, R ² represents C _1-4 alkyl, n ₁ and n ₂ are the same or different and represent an integer of 1 to 8, and m ₀ represents an integer of 1 to 8.)
The surfactant according to claim 1, comprising a cellooligosaccharide derivative represented by the formula: It consists of a cellooligosaccharide derivative having an average particle radius of 2.5 to 250 nm, in which a hydrophobic portion composed of alkylated glucose or cellooligosaccharide and a hydrophilic portion composed of monosaccharide or oligosaccharide are bound in a block manner. Nanoparticles ,
General formula:
HBA AOR ¹
H-A-B-A-OR ¹
R ¹ -A-B-OH or
R ¹ -B-A-B-OH
(In the formula, A is a general formula:
(In the formula, R ² represents C _1-4 alkyl, and m represents an integer of 1-8.)
B is composed of a hydrophilic part obtained by removing an anomeric hydroxyl group from a pyranose type monosaccharide and removing -H from one alcoholic hydroxyl group, or a pyranose type monosaccharide. 2 or more selected from the group except for the anomeric hydroxyl group of the reducing terminal sugar residue from the oligosaccharide having a 1,4-glycoside bond and -H from the alcoholic hydroxyl group at the 4-position of the non-reducing terminal sugar residue The resulting hydrophilic part, when two or more A are contained, A may be the same or different from each other, and when two or more B is contained, B may be the same or different from each other, and R ¹ is H or C _1-4 alkyl is shown. However, the sugar chain is arranged so that the reducing end is on the right side of the page. )
Nanoparticles comprising cellooligosaccharide derivatives represented by General formula:
^{R 1 -A-B-A-} OR 1 or H-B-A-B- OH
(In the formula, A is the same or different, and the general formula:
(In the formula, R ² represents C _1-4 alkyl, and m represents an integer of 1-8.)
In a hydrophobic moiety represented, B is a hydrophilic moiety obtained by removing -H from pyranose form except anomeric hydroxyl group from a monosaccharide and one alcoholic hydroxyl group, or consists of pyranose monosaccharide 2 or more selected from the group except for the anomeric hydroxyl group of the reducing terminal sugar residue from the oligosaccharide having a 1,4-glycoside bond and -H from the alcoholic hydroxyl group at the 4-position of the non-reducing terminal sugar residue The resulting hydrophilic moiety , R ¹ represents H or C _1-4 alkyl. However, the sugar chain is arranged so that the reducing end is on the right side of the page. )
A cellooligosaccharide derivative represented by: General formula (III):
(In the formula, R ¹ is H or C _1-4 alkyl, R ² is C _1-4 alkyl, m ₁ and m ₂ are the same or different and are an integer of 1-8, n ₀ is an integer of 1-8. .)
The cellooligosaccharide derivative | guide_body of Claim 10 represented by these. Formula (IV):
(In the formula, R ² represents C _1-4 alkyl, n ₁ and n ₂ are the same or different and represent an integer of 1 to 8, and m ₀ represents an integer of 1 to 8.)
The cellooligosaccharide derivative | guide_body of Claim 10 represented by these.