WO2007026883A1

WO2007026883A1 - POLYMER INCLUSION COMPOUND OF α-1,4-GLUCAN AND PROCESS FOR PRODUCTION THEREOF

Info

Publication number: WO2007026883A1
Application number: PCT/JP2006/317359
Authority: WO
Inventors: Mitsuru Akashi; Toshiyuki Kida; Takashi Minabe; Takeshi Takaha; Junichi Takahara
Original assignee: Osaka University; Ezaki Glico Co., Ltd.; Sanwa Cornstarch Co., Ltd.
Priority date: 2005-09-02
Filing date: 2006-09-01
Publication date: 2007-03-08

Abstract

Disclosed is a novel amylose inclusion compound produced by direct inclusion of a polymeric substance in the hollow interior of an amylose host molecule, more specifically an inclusion compound produced from an α-1,4-glucan host molecule having a partially modified hydroxyl group and a polymeric guest substance. Also disclosed is a process for production of the inclusion compound.

Description

Specification

Polymer inclusion compound of α-1,4_glucan and process for producing the same

Technical field

The present invention relates to a polymer inclusion complex of α-1,4-glucan and a method for producing the same.

Background art

[0002] Molecular recognition capability is a very important function in developing various functional materials. As a means for molecular recognition, the ability to form inclusions is attracting attention. Inclusion is a phenomenon in which a molecule with a cavity called a host molecule takes a guest substance into the cavity. A substance in which a guest substance is incorporated into a host molecule is called an inclusion compound.

[0003] Examples of host molecules include amylose-1,4 glucan). Amylose is a polymer in which many glucoses are linearly bound, has a helical molecular structure, and encloses a guest substance in a hollow interior.

[0004] There are many reports on complexes of amylose and low molecular weight compounds (for example, Non-Patent Document 1). However, for the complex of amylose and polymer, Kadokawa et al. Use poly (tetramethylene oxide) or poly (ε-force prolatatone) as a template to wrap around the polymer chain. An example is known in which an inclusion complex of amylose and a polymer is obtained by enzymatic polymerization of glucose 1-phosphate to amylose (Non-Patent Document 2). There is no known example in which a polymer substance is directly encapsulated in the hollow interior. Due to the strong helical structure based on the intramolecular hydrogen bond of amylose, it is considered difficult to incorporate the polymer into the amylose guest substance.

Non-Patent Document 1: Starch Science No. 34 No. 1 p. 49-57 (1987)

Non-Patent Document 2: J. Kadokawa et al. Chem. Eur. J. 2002, 8, 3321.

Disclosure of the invention

Problems to be solved by the invention

[0005] The present invention has been made in view of the above circumstances, and a novel amylose inclusion complex obtained by directly including a polymer substance in the hollow interior of an amylose host molecule and the production thereof It aims to provide a method.

Means for solving the problem

That is, the present invention provides an inclusion complex comprising an α-1,4-glucan host molecule partially modified with a hydroxyl group and a polymeric guest substance, and a method for producing the same.

[0007] "α 1, 4 glucan" is a saccharide having D glucose as a constituent unit in the present specification, and having at least two saccharide units linked only by a 4-darcoside bond. Say. α-1,4-glucan is a linear molecule. α-1,4 glucan is also called linear glucan. The number of sugar units contained in one molecule of α-1,4 glucan is called the degree of polymerization. In this specification, the term “degree of polymerization” refers to the weight average degree of polymerization unless otherwise specified. In the case of α-1,4-glucan, the weight average degree of polymerization is calculated by dividing the weight average molecular weight by 162.

[0008] In the present specification, the "host molecule" refers to a compound having a molecular structure that forms a void, such as α1,4 glucan-cyclodextrin.

[0009] "Guest material" in this specification is uniquely determined by the interaction of ionic bonds, hydrogen bonds, hydrophobic interactions, van der Waals forces, etc. with respect to the voids of host molecules. A substance that can bind specifically.

[0010] “Inclusion” refers to a state in which a host molecule incorporates a guest substance through a specific bond whose structure can be uniquely determined, and the bond is indefinite, such as aggregation or adsorption to the surface. Cannot be represented by the structure of! /, Excluding non-specific states.

[0011] The "inclusion complex" refers to a substance in which a host molecule and a guest substance are integrated by inclusion, and an aggregate thereof. Excludes non-specific agglomerates, mere mixed 'dispersions, and those covered by a film such as a capsule.

[0012] α-1, 4 glucan is a genolecan phosphorylase (a- glucan phosphorylase EC 2.

Can be synthesized using 4.1.1). An example of enzymatic synthesis of α-1,4 glucan using glucan phosphorylase is a method in which glucan phosphorylase is allowed to act on glucose-1-phosphate (G-1-—) as a substrate in the presence of maltooligosaccharide. (GP method).

[0013] Another example of enzymatic synthesis of α-1,4-gnolecan using glucan phosphorylase is An example is a method in which sucrose phosphorylase and glucan phosphorylase are allowed to act simultaneously on a substrate using SP (GP-GP). This method has the advantage that inexpensive sucrose can be used as a raw material because glucan phosphorylase acts on G-1-P generated by the action of sucrose phosphorylase to synthesize 1,4 glucan.

[0014] The α 1,4-glucan obtained by any of the above methods is a completely linear molecule without any branched structure of natural starch, and therefore has no branched structure. Easy to spiral structure. In addition, the molecular weight can be arbitrarily controlled by the reaction conditions, and the molecular weight distribution is very narrow.

[0015] The molecular weight (weight average molecular weight) of α1,4-glucan synthesized using glucan phosphorylase can be arbitrarily controlled within the range of about 5,000 to several millions. The molecular weight distribution (MwZMn) is 1.25 or less, which is much narrower than that of natural amylose. In the present invention, those having a weight average molecular weight of 10,000 to several hundred thousand, preferably about 20,000 to 70,000 are suitable. In terms of the degree of polymerization, those having a weight average degree of polymerization of about 20 to 400, preferably about 40 to 140 are suitable.

In the present specification, 4-glucan is sometimes simply referred to as “amylose”.

There are two types of α-1,4 glucans: natural amylose and enzymatically synthesized amylose. Natural amylose is contained in starch in about 20% and can be prepared by amylose selective precipitation using butanol. However, this natural amylose has various problems when actually used as a host molecule. Since natural amylose has a branched structure that is not completely a linear structure, it cannot exhibit an excellent inclusion ability. Natural amylose is a mixture of amylose having a wide degree of polymerization, and amylose having a specific degree of polymerization cannot be obtained. Enzymatic amylose, on the other hand, is an α-1,4-glucan synthesized using an enzyme and has a completely straight chain structure without a branched structure. Is possible. In the present invention, it is preferable to use enzyme-synthesized amylose.

[0017] In the α1,4-glucan in the present invention, the hydroxyl group is partially modified. The modification of the hydroxyl group is synonymous with the substitution of the hydrogen atom of the hydroxyl group with a substituent.

[0018] The position of the hydroxyl group to be modified is 2, 3, or 6 position, preferably 2, 3 position. 6th It is known that the amylose helical structure is disrupted when the hydroxyl group is modified (Hui et al. Makromol. Chem. 1988, 189, 1287.). It is done.

[0019] The substituent is an alkyl group such as a methyl group or an ethyl group, preferably a methyl group, a carboxymethyl group, or an acetyl group.

[0020] "Partial" means that all of the hydroxyl groups contained in α-1,4-glucan are not substituted with substituents, preferably hydroxyl groups at the 2, 3, 6 positions, more preferably It is substituted with 3 to 20 mol% of the hydroxyl group at the 2nd and 3rd positions. If the substitution ratio is lower than 3 mol%, it is difficult to form inclusion complex with the polymer guest substance due to the strong helical structure based on the intramolecular hydrogen bond of amylose, and the ratio is 20%. When it is larger than mol%, the intramolecular hydrogen bond of amylose is weakened, it becomes difficult to form a helical structure, and it becomes difficult to form an inclusion compound with a polymeric guest substance.

[0021] The method for producing an α-1,4-glucan host molecule partially modified with a hydroxyl group is as follows. When it is modified with an alkyl group, a halogenated alkyl dialkyl sulfate or phosphoric acid is used in a solvent or in the absence of a solvent under alkaline conditions. This can be done by the action of an alkylating agent such as trimethyl. Examples of alkyl halides include methyl iodide, acetyl iodide, methyl bromide, acetyl bromide, methyl chloride, ethyl chloride and the like. Examples of dialkyl sulfuric acid include dimethyl sulfuric acid and jetyl sulfuric acid.

[0022] Modification with a carboxymethyl group can be carried out by allowing monochloroacetic acid to act under alkaline conditions in a solvent or in the absence of a solvent.

[0023] Modification with a acetyl group can be carried out by reacting an enzymatically synthesized amylose with a acetyl chloride reaction reagent such as butyl acetate, acetic anhydride, or isopropanol acetate in a solvent or without a solvent.

[0024] Selective modification of the hydroxyl groups at positions 2 and 3 of α-1,4-glucan involves the protective action of alkylation, carboxylmethylation, acetylation, etc. after the 6-position hydroxyl group has been protected. This can be achieved by chemically modifying the hydroxyl groups at positions 2 and 3 and then deprotecting the protecting group at position 6.

[0025] Specifically, the hydroxyl groups at the 2- and 3-positions of α-1,4-glucan are selectively modified with alkyl groups. To decorate, first, α-1,4-glucan is reacted with triphenylmethyl chloride in the presence of pyridine in a solvent to selectively select the trityl group at the 6-position hydroxyl group of 4-glucan. Protect with. A 1,4-glucan with a hydroxyl group at the 6-position protected with a trityl group is added with NaH in a solvent, and an alkylating agent such as a halogenated alkyl is allowed to act on it. Examples of the alkyl halide include methyl iodide and iodinated chill. Thereafter, the trityl group protecting the hydroxyl group at the 6-position is deprotected with an acid such as hydrochloric acid to obtain α-1,4-glucan in which the hydroxyl groups at the 2,3-position are selectively modified.

[0026] In the modification with a carboxymethyl group, monochloroacetic acid is allowed to act on α-1,4-glucan in which a hydroxyl group at the 6-position is protected with a trityl group in a solvent. After that, deprotection of the trityl group at the 6-position can be performed.

[0027] Modification with the acetyl group includes dissolving hi-1,4-glucan in which the 6-position hydroxyl group is protected with a trityl group in dimethyl sulfoxide, adding pyridine, and then allowing acetic anhydride to act. It can be carried out.

[0028] When the hydroxyl group is partially modified, in the alkyl group, the ratio of the degree of substitution of the substituent can be adjusted by adjusting the addition amount of the alkylating agent or the addition amount of NaH. it can. Carboxymethyl candy can be obtained by adjusting the amount of acetic acid added to black mouth acetic acid. In addition, in the case of acetylene cake, it can be carried out by controlling the amount of acetylation reagent added and the reaction time. The desired degree of substitution can also be obtained by subjecting α- 1,4-glucan having a high degree of substitution to an alkali catalyst such as sodium hydroxide or an alkali catalyst such as sodium methoxide for deacetylation. It is possible to adjust to The degree of substitution is calculated by the integration ratio in NMR measurement. The α-1,4-glucan is synthesized by trimethylsilylating all the hydrogen atoms of the hydroxyl group remaining in the α-1,4-glucan with partially modified hydroxyl groups. The ¹ H-NMR measurement can be performed to determine the integral specific force.

[0029] The polymer as the guest substance is selected from polyether (I), polyester (II), and polyamide (III) forces represented by the following structural formula.

[0030] [Chemical 1]

In the formula (I), represents a hydrogen atom, a methyl group or an ethyl group, preferably a hydrogen atom; m represents an integer of 2 to 5, preferably 3 to 4; n is 5 to 500, preferably Represents an integer of 5 to 50, more preferably 10 to 40.

[0032] [Chemical 2]

O

^H ^ O- (CH} ^ C ^ OH (π)

R _{2 In} formula (II), R represents a hydrogen atom, a methyl group or an ethyl group, preferably a hydrogen atom;

2

m represents an integer of 3 to 5, preferably 4 to 5; n is 5 to: LOOO, preferably 5 to 100

twenty two

Preferably it represents an integer of 7 to 90.

[0033] [Chemical 3]

In the formula (III), R represents a hydrogen atom, a methyl group or an ethyl group, preferably a hydrogen atom.

Three

M represents an integer of 1 to 5, preferably 3 to 5; n is 10 to 400, preferably 10 to 40

3 3

Preferably it represents an integer of 20-30.

[0035] The inclusion complex of the present invention can be obtained by dissolving an α-1,4-glucan host molecule partially modified with a hydroxyl group and a polymer guest substance in the same solvent and mixing them. .

[0036] The solvent used in the present invention is a solvent that can dissolve both the host molecule and the guest molecule, and preferably a solvent that can be dissolved at room temperature. As such a solvent, for example, water or a mixed solvent of water and an organic solvent compatible with water is used. Examples of organic solvents that are compatible with water include dimethyl sulfoxide (DMSO), ethanol, methanol, acetone, acetonitrile, isopropanol, tetrahydrofuran, dimethylformamide, and dioxane. Among them, dimethyl sulfoxide (DMSO) is highly soluble in amylose. It is excellent and preferable. As a mixed solvent of water and dimethyl sulfoxide (DMSO), for example, 99: 1 to 70:30, preferably 95: 5 to 80 of water and dimethyl sulfoxide (DMSO) is used. : Use a mixed solvent of 20 (HO: DMSO)!

2

[0037] The solvent used in the present invention may contain various inorganic salts and low molecular organic compounds as long as they do not have an inhibitory effect on the formation of the inclusion complex between the host molecule and the guest molecule. Good.

[0038] The concentration at which the host molecule is dissolved in the solvent may be about 0.1 to 30% by weight, preferably about 0.1 to 5% by weight, more preferably about 0.3 to 3% by weight. If the concentration is too low, it is difficult to obtain clathrate precipitates, and productivity is poor and not economical. If the concentration is too high, the viscosity becomes high and handling becomes difficult. Also, even if the concentration is increased to a certain extent, productivity is not improved.

[0039] The concentration at which the guest substance is dissolved in the solvent may be about 0.1 to 30% by weight, preferably about 0.1 to 5% by weight, more preferably about 0.3 to 3% by weight. If the concentration is too low, it is difficult to obtain clathrate precipitates, and productivity is poor and not economical. If the concentration is too high, the viscosity becomes high and handling becomes difficult. Also, even if the concentration is increased to a certain extent, productivity is not improved.

[0040] The mixing ratio of the host molecule and the guest molecule dissolved in the solvent is such that the polymer guest molecule constituent unit is 1 unit or more, preferably 10 units or more per 6 units of the glucan host molecule constituent unit. Like that. If there are too few polymer guest molecule constituent units, the clathrate compound cannot be precipitated.

[0041] The mixing means is not particularly limited, but may be mixed by a method such as ultrasonic irradiation, a homogenizer, a propeller stirrer, a magnetic stirrer, a static mixer, or a combination thereof. Mixing by ultrasonic irradiation is preferred.

[0042] The temperature of the solution in the mixing step is 20 to 100 ° C, preferably 50 to 90 ° C. If the temperature is too high, the operation becomes extremely difficult. If the temperature is too low, the amylose has a strong herritus and no inclusion complex formation occurs. The longer the mixing time, the better, but it may be about 1 to 60 minutes, preferably about 5 to 30 minutes.

[0043] It is preferable that the mixed solution is mixed at the above temperature and time and then allowed to stand. The standing condition may be about 1 to 24 hours under normal temperature and pressure.

[0044] When the produced inclusion complex is obtained as a precipitate, the precipitate is centrifuged, The clathrate is recovered by a method such as filtration, and the precipitate is not dissolved, but may be purified by washing the precipitate with an appropriate solvent that dissolves only the guest polymer. When it is obtained in a state of being dissolved in a mixed solvent rather than as a precipitate, an inclusion compound is precipitated by adding a solvent such as ethanol or methanol to the mixture, and then the precipitate is The clathrate compound may be purified and collected by collecting and washing in the same manner as described above.

[0045] The force of obtaining an inclusion complex composed of an α-1,4 glucan host molecule partially modified with a hydroxyl group and a polymeric guest substance is characterized by the inclusion compound by X-ray diffraction. It can be confirmed from the spectrum obtained by NMR measurement that a typical peak has occurred.

The invention's effect

[0046] A novel amylose-polymer inclusion complex and a method for producing the same were provided.

[0047] [Example]

2. Synthesis of 3-methylated amylose

[Chemical 4]

2,3-position methylated amylose

[0048] 1) Tritylation of 6-position hydroxyl group of amylose

Amylose (molecular weight Mw = 21360 Mw / Mn = l. 05) 1. Og (4. 7 X 10 " ⁵ mol, 6.2 X 10" ³ mol / glucose unit) and triphenyl-noremethinolect in 30 mL of pyridine under nitrogen atmosphere After adding 5.3 g (l. 9 X 10 ^_2 mol) of mouthride and refluxing at 110 ° C for 4 hours, the temperature of the system was lowered to 60 ° C. Methanol was added until the solution became opaque, the temperature was further lowered to room temperature, and 400 mL of methanol was added. The precipitated pale yellow precipitate was collected by filtration, washed with methanol, and dried under reduced pressure. Pale yellow solid, yield 2.25 g, yield 90%

[0049] 2) 6-O-Trityl amylose methyl hydroxyl group

Under nitrogen atmosphere, tetrahydrofuran (THF) lOmL was charged with 6-O-tritylamylose 0.60 g (l. 1 X 10 " ⁵ mol, 1.5 X 10 ^_3 mol / glucose unit) and the prescribed amount of NaH below. After stirring at 0 ° C for 1 hour, 0.56 mL (9. OX 10 ^_3 mol) of yowi-methyl was added and stirred for 1 hour at room temperature. It was collected by filtration, washed with methanol, and dried under reduced pressure.

[0050] Amount of NaH added (2.2 X 10 " ³ g, 9.0 X lO 'mol): 3% methylation of hydroxyl groups at 2 and 3 positions

NaH-added amount (5.8 X 10 " ³ g, 2.4 X lO 'mol): When 8% methylation of the 2nd and 3rd hydroxyl groups NaH added amount (1.4 X 10— ² g, 6.0 X lO 'mol): 20% methylation of hydroxyl groups at the 2nd and 3rd positions

Addition amount of NaH (2.4 X 10 " ² g, 9.9 X lO 'mol): When 33% methylates hydroxyl groups at 2 and 3 positions

NaH-added amount (0.14 g, 6.0 X lO 'mol): When 100% methylation of the hydroxyl groups at the 2nd and 3rd positions [0051] 3) Methylation at the 2nd and 3rd positions Conversion

THF20mL 2, 3-position methylation 6- O-trityl amylose (1. 1 X 10 ^_5 mol) of Karoe to添Ka卩a 0. 2M HC1 solution in methanol 20 mL, was 2晚stirred at room temperature. Na CO (6.2

twenty three

X 10 ^_3 mol) aqueous solution, and the solvent was distilled off. The obtained solid was washed with hexane and dialyzed against water using a dialysis membrane having a molecular weight fraction of 3500 for 3 days.

[0052] [Table 1]

Synthesis of 2,3-position methylated amylose

Raw material of amylose

[NaH] / [OH] Yield (%) Product * Molecular weight (Mw) Degree of methylation (%)

0.03 30 3 MA2-3

0.083 82 8 MA2-8

21, 360 0.20 35 20 MA2-20

0.33 86 33 A2-33

2.0 64 100 MA2-100

0.083 64 8 MA5-8

49,680 0.20 80 20 MA5-20

0.83 35 50 MA5-50

* For example, the raw material amylose has a molecular weight of 21,360, and the methylation degree of the hydroxyl group at positions 2 and 3 is 3%. This methylated amylose is expressed as MA2-3, and the molecular weight of amylose is 9,680, and the methylated amylose with 8% methylation at the 2- and 3-position hydroxyl groups is expressed as MA5-8.

[0053] A study of clathrate formation in solution

The presence or absence of inclusion complex of 2,3-position methyl amylose and various guest polymers was evaluated by whether or not a precipitate was formed after mixing each polymer solution (dispersion).

[0054] Structure of host polymer and guest polymer

[Chemical 5]

Host polymer (methylated amylose)

R = H or Me

[Chemical 6]

Guest polymer

PTHF nn == 9 y ,, 1 Ί 4 ,, 40U PCL n = 1 1, 1 7

[0055] MA2-3-MA2-100, MA5-8-MA5-50 were used as host polymers. As the guest polymer, poly (tetramethylene oxide) (PTHF) and poly (force prolatatone) (PC L) were used.

[0056] Methyl amylose in Table 1, PTHF and PCL having different degrees of polymerization were dissolved in a solution 21111 ^ of DMSO: H 0 = 1: 9 so as to have final concentrations shown in Table 2, respectively. 15 at 60 ° C Ultrasonic waves were irradiated for 1 minute, and after standing at room temperature for a while, the presence or absence of precipitation was observed. For those in which precipitation was confirmed, the precipitate was collected by centrifugation, and the resulting precipitate was washed twice with methanol and once with water, freeze-dried overnight, and then weighed. The results are summarized in Table 2.

[0057] [Table 2]

Mixing ratio of host polymer and guest polymer

Host polymer-Guest polymer-Precipitation Yield (mg) mol / L .W, mol /

MA 2-3 3.0> (10— ⁴ 2900 3.0X10— ³ Yes 8,8

MA 2-8 3.0> 0 one ⁴ 2900 3.0X10- ³ Yes 5.7

MA 2-20 3.0><10 one ⁴ 2900 3.0X10- ³ Yes 1.5

PTHF

MA 2-33 3.0> (1CT ⁴ 2900 3.0x1 T ³ ―

MA 2-50 3.0> O-4 2900 3.0x1CT ³ None ―

MA 2-100 3.0> 2900 3.0x10— 3 ―

MA 5-8 1.3> 2900 3.0x10— ³ 8.3

MA 5-20 1.3> 0 1 ⁴ PTHF 2900 3,0x1Cr ³ Yes 5.3

^{MA 5-50 1.3> (10- 4 2900} 3.C 0- 3 No -

[0058] 丄 H-NMR measurement

Dissolve 5 mg of the resulting precipitate in 0.75 mL of dimethylsulfoxide d and add JEOL (day

6

¹ H-NMR was measured at 20 ° C with this electron) Hen NM-GX 400 (400MHz).

[0059] FIG. 1 shows the ¹ H-NMR ^ vector of the inclusion compound of MA2-8 and PTHF.

The ¹ H-NMR ^ vector of the inclusion compound of MA5-20 and PTHF is shown in FIG.

The ¹ H-NMR ^ vector of the inclusion compound of MA28 and PCL is shown in FIG.

[0060] [Chemical 7] 2,3-position methylated amylose

[0061] In FIG. 1, the integral ratio of the 1-position proton (H) of the glucose unit in MA2-8 to the j8-position hydrogen proton (H) of PTHF is 5.0: 4.0. 1 unit of glucose H

β 1

There is one, and there are four HF in the PTHF1 unit. From the integral ratio of each proton, Μ

β

It is suggested that Α2-8 includes PTHF1 unit with 5 glucose units.

[0062] Similarly, Fig. 2 Force et al. 5-20 suggests that the PTHF1 unit is clathrated with 4.5 glucose units. In FIG. 3, the integral ratio of the 1-position proton (H) of the glucose unit in MA2-8 to the ε-position hydrogen proton (Η ε) of PCL is 7.1: 2.0. There is one Η per glucose unit and two ε ε per PCL1 unit. For this reason, it is suggested that 82-8 includes the PCL1 unit with a 7.1-course unit.

[0064] X-ray diffraction method (XRD measurement)

The obtained precipitate was placed on a glass substrate, and XRD measurement was performed at room temperature using RINT InPlane / ultraX18SAXS-IP manufactured by Rigaku.

Measurement conditions: A slit having a scanning speed of 0.5 ° Zmin and a width of 0.3 mm was used.

FIG. 4 shows an XRD pattern diagram of PTHF-MA2-8 inclusion compound.

Figure 5 shows the XRD pattern of the PTHF-MA5-20 clathrate compound.

Figure 6 shows the XRD pattern of the PCL-MA2 8 clathrate compound.

[0066] The XRD pattern of the inclusion complex is different from that of each methylated amylose and guest polymer (PTHF, PCL), and the characteristic peaks of the inclusion complex from XRD (Fig. 3, 2 Θ = 12 5 °, 2 0 = 20. 1 °) was observed. This is almost the same as the results previously reported by Kadokawa et al.

[0067] Synthesis of 2.3-acetylated amylose

1) Acetylation of hydroxyl groups 2 and 3 of 6-0-trityl amylose

experimental method:

Under nitrogen atmosphere, 6-O trityl amylose 0. lg (l. 8 X 10 " ⁶ mol, 2.5 X 10 ^_4 mol / glucose unit) was dissolved in 2.5 mL of dehydrated dimethyl sulfoxide (DMSO). 2.5 mL of dehydrated pyridine and 40 mL of acetic anhydride were added, and the mixture was stirred at room temperature for 12 hours, 50 mL of methanol was added, and the precipitated pale yellow solid was collected by filtration, washed with methanol, and dried under reduced pressure.

[0068] 2) Deacetylation of 6-O trityl amylose at 2- and 3-position acetylation

To 20 mL of THF was added 2,3-position acetylated 6-O trityl amylose (1.1 X 10 ^_5 mol), 20 mL of 0.2 M HC1 methanol solution was added, and the mixture was stirred at room temperature for 2 hours. Na CO (6.

twenty three

2 X 10 ^_3 mol) aqueous solution, and the solvent was distilled off. The obtained solid was washed with hexane and dialyzed against water for 3 days using a dialysis membrane having a molecular weight fraction of 3500. Then freeze-dry The product was obtained.

[0069] For the examination of the inclusion complex formation, 2 or 3 position acetylyl amylose having 15% acetylyl chloride was used.

[0070] Review of inclusion complex formation in solution

The presence or absence of inclusion complex formation between acetylyl amylose and guest polymer was evaluated by whether or not a precipitate was formed after mixing each polymer solution (dispersion).

[0071] Structure of host polymer and guest polymer

[Chemical 10]

Host polymer

2,3-position acetylated amylose

R = H or COCH ₃

[Chemical 11]

Guest polymer

[0072] As the host polymer, 2,3-position acetylated amylose was used. As the guest polymer, poly (tetramethylene oxide) (PTHF) (molecular weight Mw = 2900) was used.

[0073] DMSO: H 0 = 1 so that the final concentrations of 2,3-position acetylated amylose and PTHF are 3.0 X 10 " ⁴ mol / L and 2.2 X 10 ^_3 mol / L, respectively. : Dissolved in 2 mL of 9 solution . The mixture was allowed to stand at 60 ° C for 12 hours and then at room temperature for 12 hours to obtain a precipitate. The precipitate was collected by centrifugation. The resulting precipitate was washed twice with methanol and once with water, freeze-dried overnight, and then analyzed using '-NMR and XRD.

[0074] 丄 H-NMR measurement

Dissolve 5 mg of the resulting precipitate in dimethylsulfoxide d 0.75 mL and add JEOL (Japan

6

¹ H-NMR was measured at 20 ° C with an electron) NM-GX 400 (400 MHz).

[0075] FIG. 7 shows the ¹ H-NMR spectrum of the inclusion complex of 2,3-position acetylated amylose and PTHF.

[0076] [Chemical 12]

2,3-position acetylated amylose

R = H or COCH ₃

[Chemical 13]

PTHF

In Fig. 7, the integral ratio of the 1-position proton (H) of the glucose unit in acetylated amylose to the j8-position hydrogen proton (H) of PTHF is 2.8: 4.0. 1 unit of glucose

β

There is one persimmon, and there are four persimmons per PTHF1 unit. Integration of each proton

1 β

The ratio suggests that 2.8 glucose units enclose PTHF1 units. [0078] X-ray diffraction method (XRD measurement)

FIG. 8 shows an XRD pattern diagram of the acetylated amylose PTHF inclusion complex.

[0080] The XRD pattern of the inclusion complex is different from that of the acetylated amylose and guest polymer (PTHF) alone, and the characteristic peaks of the amylose inclusion complex (Figs. 8, 20 = 12.9 °, 2 0 = 20. 2 °) was observed.

[0081] Synthesis of 2,3-position carboxymethylated amylose

1) Carboxymethylation of hydroxyl groups 2 and 3 of 6-0-tritylamylose

experimental method:

6-O trityl amylose 0. lg (l. 8 X 10 " ⁶ mol, 2.5 X 10 ^_4 mol / glucose unit) was dissolved in 2.5 mL of dehydrated dimethyl sulfoxide (DMSO) under a nitrogen atmosphere. Add 100 mL (l. 0 6 X 10 ^_3 mol) of monoclonal acetic acid while stirring and stirring 5 mL of NaOH aqueous solution, and stir for 24 hours at room temperature. It was collected by filtration, washed with methanol, and dried under reduced pressure.

[0082] 2) Detritylation of 6-O trityl amylose at 2- and 3-position carboxymethylation

Add 2,3-position carboxymethylated 6-O trityl amylose (1.1 X 10 " ⁵ mo 1) to 20 mL of THF, add 20 mL of 0.2 M HC1 methanol solution, and stir at room temperature for 2 hours. C

2

The mixture was neutralized with an aqueous solution of O (6.2 X 10 ^_3 mol) and the solvent was distilled off. The resulting solid with hexane

Three

After washing, the mixture was dialyzed against water for 3 days using a dialysis membrane having a molecular weight fraction of 3500. Thereafter, the product was obtained by freeze-drying.

[0083] For the examination of the inclusion complex formation, carboxymethyl amylose having a carboxymethyl glycerol concentration of 3% was used.

[0084] Review of inclusion complex formation in solution

The presence or absence of inclusion complex formation between carboxymethyl amylose and guest polymer was evaluated by whether or not a precipitate was formed after mixing each polymer solution (dispersion).

[0085] Structure of host polymer and guest polymer [Chemical 14]

Host polymer

2,3-carboxymethylated amylose

R = H or CH ₂ C〇OH

[Chemical 15]

Guest polymer

PTHF Π = 40

[0086] As the host polymer, 2,3-position carboxymethylated amylose was used. As the guest polymer, poly (tetramethylene oxide) (PTHF) (molecular weight Mw = 2900) was used.

[0087] Carboxymethylated amylose and PTHF were mixed with DMSO: H 0 = 1: 9 so that the final concentrations were 3.0 X 10 " ⁴ mol / L and ^2.2 X 10 ^_3 mol / L, respectively. Dissolve in 2 mL of solution

2

It was. After standing at 60 ° C for 12 hours, the mixture was further allowed to stand at 20 ° C for 12 hours to obtain a precipitate. The precipitate was collected by centrifugation. The resulting precipitate was washed twice with methanol and once with water, freeze-dried overnight, and then analyzed using 1 H-NMR and XRD.

[0088] 丄 H—NMR measurement

6

¹ H-NMR was measured at 20 ° C with an electron) NM-GX 400 (400 MHz).

[0089] FIG. 9 shows the ¹ H-NMR ^ vector of the inclusion complex of 2,3-position carboxymethylated amylose and PTHF. [0090] [Chemical 16]

2, 3-position Lupoxymethylated Amylose

R = H or CH ₂ COOH

[Chemical 17]

PTHF

[0091] In FIG. 9, the integral ratio of the 1-position proton (Η) of the glucose unit in carboxymethylated amylose and the j8-position hydrogen proton (Η) of PTHF is 3.4: 4.0. Glucose 1

β

There is one Η in the unit, and there are four Η in the PTHF1 unit. Each proto

1 β

The integral ratio suggests that carboxymethylated amylose clathrates PTHF1 units with 3.4 glucose units.

[0092] X-ray diffraction method (XRD measurement)

[0093] Fig. 10 shows the XRD pattern of the carboxymethylated amylose PTHF inclusion complex. [0094] The XRD pattern of the inclusion complex is different from that of carboxymethylated amylose and guest polymer (PTHF) alone, and the characteristic peaks of the amylose inclusion complex (Fig. 10, 2 Θ = 12.9 ° , 2 0 = 20. 1.) was observed.

Industrial applicability

[0095] By selectively removing the guest polymer from the ultra-thin film obtained by laminating the clathrate compound of the present invention on the substrate, a porous amylose ultra-thin film storing its three-dimensional structure precisely is obtained. Can be produced. Since only the polymer having a specific structure (molecular weight, stereoregularity, monomer order) can enter the pores, it can be applied as a separation analysis material that can identify the polymer structure with high accuracy. The hollow capsule formed from the ultrathin film of this amylose monopolymer inclusion complex can be used as a drug carrier that can control the release of the encapsulated drug by removing the guest polymer.

Brief Description of Drawings

[0096] [FIG. 1] ¹ H-NMR ^ vector diagram of inclusion compound of MA2-8 and PTHF.

[Fig. 2] ¹ H-NMR ^ vector diagram of inclusion compound of MA5-20 and PTHF.

[Fig. 3] ¹ H-NMR ^ vector diagram of inclusion compound of MA2-8 and PCL.

FIG. 4 is an XRD pattern diagram of PTHF—MA2 8 inclusion compound.

FIG. 5: XRD pattern diagram of PTHF-MA5-20 clathrate compound.

FIG. 6: XRD pattern diagram of PCL—MA2 8 inclusion compound.

FIG. 7 is a ¹ H-NMR spectrum of an inclusion complex of 2,3-position acetylated amylose and PTHF.

[FIG. 8] XRD pattern diagram of 2,3-position acetylated amylose PTHF inclusion complex.

[Fig. 9] ¹ H-NMR spectrum of an inclusion complex of 2,3-position carboxymethylated amylose and PTHF.

[Fig. 10] XRD pattern of the 2,3-position carboxymethylated amylose PTHF inclusion complex.

Claims

The scope of the claims

[I] An inclusion compound comprising an α-1,4 glucan host molecule partially modified with a hydroxyl group and a polymeric guest substance.

[2] The clathrate according to claim 1, wherein the modification is that the hydrogen atom of the hydroxyl group is substituted with a substituent.

[3] The inclusion compound according to claim 2, wherein the substituent is selected from an alkyl group having 1 to 2 carbon atoms, a carboxymethyl group, or a acetyl group.

[4] The clathrate according to any one of claims 1 to 3, wherein the hydroxyl group to be modified is a hydroxyl group at the 2- and 3-positions.

[5] The clathrate compound according to any one of claims 1 to 4, wherein the hydroxyl group is substituted with a substitution degree of 3 to 20%.

[6] The clathrate compound according to any one of claims 1 to 5, wherein the polymer is selected from polyether forces.

[7] The inclusion of the host molecule and the guest substance, including the step of dissolving and mixing the α-1,4 glucan host molecule partially modified with a hydroxyl group and the polymer guest substance in the same solvent Compound manufacturing method.

[8] The method for producing an clathrate compound according to [7], wherein the modification is that a hydrogen atom of a hydroxyl group is substituted with a substituent.

[9] The method for producing an inclusion compound according to claim 8, wherein the substituent is selected from an alkyl group having 1 to 2 carbon atoms, a carboxymethyl group, or a acetyl group.

[10] The method for producing an inclusion compound according to any one of [7] to [9], wherein the hydroxyl group to be modified is a hydroxyl group at the 2- or 3-position.

[II] The method for producing an clathrate compound according to any one of claims 7 to: wherein the hydroxyl group is substituted with a substitution degree of 3 to 20%.

12. The method for producing an inclusion compound according to any one of claims 1 to 11, wherein the polymer is selected from polyether forces.