WO2013172436A1

WO2013172436A1 - Novel glycosylated compound of crocin, method for producing same, and use for same

Info

Publication number: WO2013172436A1
Application number: PCT/JP2013/063730
Authority: WO
Inventors: 裏出　良博; 志力黄; 浩介有竹; 正山　征洋; 米谷　俊
Original assignee: 公益財団法人大阪バイオサイエンス研究所
Priority date: 2012-05-18
Filing date: 2013-05-16
Publication date: 2013-11-21

Abstract

The invention pertains to glycosylated crocin compounds obtained by condensing glucose or glucose oligomer to one or more hydroxyl groups of crocin represented by formula (1) as novel glycosylated compounds of crocin, or a mixture thereof; a method for the production thereof; and new uses for glycosylated crocin compounds.

Description

Novel sugar addition compound of crocin, method for producing the same, and use thereof

The present invention relates to a novel sugar addition compound of crocin and a method for producing the same. Furthermore, this invention relates to the new use of the said crocin sugar addition compound.

In the modern society, the number of people who suffer from insomnia due to stress and 24-hour lifestyle tends to increase. According to a survey by the Ministry of Health, Labor and Welfare, the percentage of people who are in trouble with insomnia has exceeded 20%, which is a serious problem. For this reason, the demand for drugs that improve insomnia is expected to increase in the future, but many of the currently used sleeping pills or sleep inducers have side effects such as headaches, discomfort, and physical dependence after waking, not necessarily It is hard to say that it is a medicine that brings natural sleep. Accordingly, there is a demand for the development of drugs that can provide more comfortable sleep.

For this purpose, the present inventors have found that crocin contained in saffron or gardenia has an action of promoting sleep, an action of significantly extending non-REM sleep, which is an indicator of deep sleep, and a natural and comfortable sleep. Has already been found and published (see Patent Document 1, Non-Patent Document 1, etc.).

As shown in the following formula (I), the crocin is gentiobiose (a disaccharide in which two glucose molecules are bonded via a 1-6 bond) to two carboxyl groups of crocetin (Patent Document 2), which is known to have a sleep-improving action. It is a sugar adduct of crocetin with condensed residues, and it has been found that the sleep promoting action is 10 times higher than crocetin (Non-patent Document 1).

JP 2008-273939 A JP 2010-246357 A

An object of the present invention is to provide a novel compound that can be an active ingredient of a sleep improving agent or sedative agent that brings about safe and natural sleep, and a method for producing the same. More specifically, the object of the present invention is to provide a novel compound that is superior in oral absorbability compared to the aforementioned crocin (I) and that can exert sleep-improving action or sedation action with a small dose, and production thereof Is to provide a method. Another object of the present invention is to provide a novel compound excellent in stability in an aqueous solution and / or stability against light irradiation as compared with the aforementioned crocin (I), and a method for producing the same. . Furthermore, the objective of this invention is providing the use as a sleep improving agent or a sedative of the said compound.

The inventors of the present invention have made extensive studies in order to achieve the above-mentioned object. As a result, a sugar adduct of crocin obtained by enzymatic treatment of crocin (I), specifically crocin (I), is provided with a sugar donation. Enzymatic sugar adducts of crocin obtained by the action of glucose transferase in the presence of the body are highly absorbable for oral administration and have a lower dose than oral administration of crocin (I) itself. It has been found that an excellent sleep-improving effect or sedative effect suitable for the above purpose can be obtained by administration, and that it is more stable in aqueous solution and more stable to light than crocin (I). It was confirmed that the sugar adduct was useful as an oral sleep improving agent or sedative, and as an oral composition (for example, food and drink) that naturally promotes sleep and sedation.

The present invention has been completed based on such findings, and includes the following embodiments.

(A) Crosin sugar addition compound or a mixture thereof (A-1) A crocin sugar addition compound or a mixture thereof obtained by condensing glucose or a glucose oligomer to one or more hydroxyl groups of crocin (I) represented by the following formula:

(A-2) A crocin sugar addition compound or a mixture thereof according to (A-1) represented by the following formula (II):

(In the formula, Glc is a glucose residue, and n ₁ to n ₆ are the same or different and each represents an integer of 0 to 20, except that all of n ₁ to n ₆ are 0.) .

(A-3) The “crocin sugar addition compound” described in (A-2) is a compound in which n ₁ is 1 or 2 and n ₂ to n ₆ are 0 in formula (II); The crocin sugar addition compound or mixture thereof according to (A-2), wherein the “mixture” described in A-2) contains the crocin sugar addition compound.

(A-4) According to any one of (A-1) to (A-3), which is produced by a method in which crocin (I) is reacted with glucose residue transferase in the presence of a sugar donor The crocin sugar adduct compound described or a mixture thereof.

(A-5) The crocin sugar addition compound or mixture thereof according to any one of (A-1) to (A-4), wherein the sugar donor is a glucose source.

(A-6) The crocin according to (A-5), wherein the glucose source is at least one selected from the group consisting of glucose, maltose, amylose, amylopectin, starch, starch liquefaction, starch saccharification, and cyclodextrin Sugar addition compounds or mixtures thereof.

(A-7) The crocin sugar addition compound or mixture thereof according to any one of (A-1) to (A-6), wherein the glucose residue transferase is transglucosidase.

(A-8) The crocin sugar addition compound or mixture thereof according to (A-7), wherein the transglucosidase is cyclodextrin glucanotransferase.

(B) Production method of crocin sugar addition compound or mixture thereof (B-1) Crocin (I) represented by the following formula:

And a method for producing a crocin sugar adduct compound represented by the following formula (II) or a mixture thereof, comprising the step of allowing a glucose residue transferase to act in the presence of a sugar donor:

(B-3) The production method according to (B-1) or (B-2), wherein the sugar donor is a glucose source.

(B-4) Described in (B-3), wherein the glucose source is at least one selected from the group consisting of glucose, maltose, amylose, amylopectin, starch, starch liquefaction, starch saccharification, and cyclodextrin Production method.

(B-5) The production method according to any one of (B-1) to (B-4), wherein the glucose residue transferase is transglucosidase.

(B-6) The production method according to (B-5), wherein the transglucosidase is a cyclodextrin glucanotransferase.

(C) Use of crocin sugar addition compound or a mixture thereof (C-1) A composition containing the crocin sugar addition compound or a mixture thereof according to any one of (A-1) to (A-8).

(C-2) The composition according to (C-1), which is an oral composition.

(C-3) The composition described in (C-1) or (C-2), which is a pharmaceutical, a quasi-drug, or a food or drink.

(C-4) The composition according to any one of (C-1) to (C-3), which is a sleep improving agent or a sedative.

(C-5) The composition according to (C-4), wherein the sleep improving agent improves sleep based on increasing non-REM sleep time.

(C-6) The composition according to (C-4), wherein the sedative suppresses the amount of behavior via a histamine receptor.

(C-7) A crocin sugar adduct compound or a mixture thereof according to any one of (A-1) to (A-8), a test animal (including a human), preferably an animal having a sleep disorder or A method for improving sleep or sedation for an animal, comprising the step of administering to the animal to be sedated.

(C-8) The method according to (C-7), wherein the sleep improvement method is a method of improving sleep based on increasing non-REM sleep time.

(C-9) The method according to (C-7), wherein the sedation method is a method of suppressing an action amount via a histamine receptor.

(C-10) The method according to any one of (C-7) to (C-9), wherein the animal is a non-human animal.

(C-11) (A-1) to (A-8) used for improving or sedating sleep in test animals (including humans), preferably animals having sleep disorders or animals to be sedated Or a mixture thereof.

(C-12) Use of a crocin sugar addition compound or a mixture thereof according to any one of (A-1) to (A-8) for producing a sleep improving agent or a sedative.

(D) A method for enhancing the oral absorbability of crocin (D-1) A glucose residue transferase is allowed to act on crocin represented by the above formula (I) in the presence of a sugar donor. A method for enhancing the oral absorbability of crocin, comprising the step of producing a crocin sugar adduct compound represented by Note that this method is a method for modifying crocin to improve the physical property (characteristic) of crocin absorption in the body, and is different from medical practice for animals including humans.

(D-2) The crocin sugar addition compound represented by formula (II) is a compound in which n ₁ is 1 or 2 and n ₂ to n ₆ are 0 in formula (II) (D-1 The manufacturing method described in the above.

(D-3) The method according to (D-1) or (D-2), wherein the sugar donor is a glucose source.

(D-4) Described in (D-3), wherein the glucose source is at least one selected from the group consisting of glucose, maltose, amylose, amylopectin, starch, starch liquefaction, starch saccharification, and cyclodextrin Method.

(D-5) The method according to any one of (D-1) to (D-4), wherein the glucose residue transferase is transglucosidase.

(D-6) The method according to (D-5), wherein the transglucosidase is a cyclodextrin glucanotransferase.

ADVANTAGE OF THE INVENTION According to this invention, the crocin sugar addition compound which is a novel compound which accelerates | stimulates sleep by ingestion, and can obtain a natural and comfortable sleep, or a novel compound which promotes sedation, and its mixture can be provided. . Hereinafter, in this specification, a crocin sugar adduct that means a single compound and a mixture of the various crocin sugar adducts may be collectively referred to as “crocin sugar adduct”.

The crocin sugar addition compound is less than crocin (Patent Document 1, Non-Patent Document 1), which is known to have a sleep improvement effect and a sedative effect, and, similarly to crocin, significantly reduces non-REM sleep, which is an indicator of deep sleep. Prolongs action and sedation.

For this reason, the crocin sugar adduct and its mixture (crocin sugar adduct) are used as components of oral compositions (pharmaceuticals, quasi-drugs, foods and drinks) that promote sleep or sedation, especially for oral sleep-improving agents or Useful as an active ingredient in sedatives.

Moreover, the crocin sugar adduct of the present invention is superior in stability in aqueous solution and / or stability to light irradiation as compared with crocin. For this reason, it is possible to supply a pharmaceutical, a quasi-drug, or a food or drink containing a crocin sugar adduct as a component to the market as a product with stable quality.

The production transition of the enzymatic transglycosylation reaction product of crocin (crocin sugar adduct) is determined by measuring the reaction mixture at the start of reaction (0hr), 1 hour (1hr), 3hr (3hr) and 6hr (6hr) after the reaction. The result confirmed by HPLCH analysis is shown (Example 1). (A) The results of evaluating the stability in aqueous solution (room temperature) for crocin (circle number 1 in the figure) and crocin sugar adduct (circle numbers 2 to 9 in the figure) over 24 hours are shown. (B) The results of evaluating the stability against light irradiation over 24 hours for crocin (circle number 1 in the figure) and crocin sugar adduct (circle numbers 2 to 9 in the figure) are shown (both are experimental example 1). . (A) About the test group The cumulative sleep amount for 6 hours after oral administration of crocin sugar adduct (25 mg / kg) for each of the control group (solvent administration group) for each wake time, REM sleep time, and non-REM sleep time The result compared with is shown. (B) About the test group After the oral administration of the crocin sugar adduct (2.5 mg / kg, 25 mg / kg), the cumulative sleep amount (REM sleep time and non-REM sleep time) for 4 hours and the crocin suspension of the comparison group The result of comparing the cumulative sleep amount (REM sleep time and non-REM sleep time) for 4 hours after intraperitoneal administration (10 mg / kg, 30 mg / kg) is shown (both are experimental example 2). In Experimental Example 2, the crocin sugar adduct was administered intraperitoneally (FIG. 4 (A)) or orally (FIG. 4 (B)), and blood was collected 15 minutes, 30 minutes and 60 minutes after administration, The result of having measured the quantity of the crocin sugar adduct, crocin, and crocetin in the blood is shown. In Experimental Example 3, various mice (C57BL / 6 male mice [shown as “C57BL / 6” in the figure]), adenosine A1 receptor gene-deficient mice [shown as “adenosine A1 receptor KO” in the figure] and Histamine H1 receptor gene-deficient mice [indicated as “Histamin H1 receptor KO” in the figure] were each orally administered with a crocin sugar adduct, and the amount of behavior in 12 hours from 19:00 to 6 o'clock (FIG. 5 ( A)) and the accumulated action amount (FIG. 5B) are measured. In addition, the result when the solvent is orally administered as a comparative control experiment is also shown. The reaction solution after 6 hours of reaction in Example 1 was subjected to HPLC, and each fraction fractionated from each peak (

circle numbers

1, 2, 3, 4 and 5 in order from the right in the bottom diagram of FIG. 1) ( The results of subjecting Fraction-1, 2, 3, 4 and 5) to the mass spectrum are shown. The mass spectra of Fraction-1, 2, 3, 4 and 5 are shown in order from the top (Experimental Example 4). FIG. 6 shows the results (13C-NMR data) of Fraction-1, 2, and 3 shown in FIG. 6 subjected to ¹³ C-NMR spectrum (solvent: dimethyl sulfoxide-d6 [DMSO-d6]). FIG. 6 shows the results (1H-NMR data) of Fraction-1, 2, and 3 shown in FIG. 6 subjected to ¹ H-NMR spectrum (solvent: DMSO-d6). The structures of Fraction-1, 2, and 3 compounds (crocin, crocin-monoglycoside, crocin-diglycoside, respectively) are shown.

(A) Crosin sugar addition compound, mixture thereof, and production method thereof The crocin sugar addition compound targeted by the present invention is obtained by condensing glucose or glucose oligomer with one or more hydroxyl groups of crocin (I) represented by the following formula: Means the compound. The target crocin includes structural isomers comprehensively included in the following formula (I) (for example, bis [(2S, 3R, 4S, 5S, 6R) -3,4,5-trihydroxy- 6-[[(2R, 3R, 4S, 5S, 6R) -3,4,5-trihydroxy-6- (hydroxymethyl) oxan-2-yl] oxymethyl] oxan-2-yl] (2E, 4E, 6E, 8E, 10E, 12E, 14E) -2,6,11,15-tetramethylhexadeca-2,4,6,8,10,12,14-heptaenedioate).

Such a crocin sugar addition compound may have different binding sites and the number of glucose or glucose oligomers, the number of glucose residues constituting the glucose oligomer, and the like. The mixture of crocin sugar addition compounds referred to in the present invention means a mixture containing two or more kinds of various crocin sugar addition compounds. As described above, in this specification, a mixture of a crocin sugar adduct compound that means a single compound and the various compounds is collectively referred to as a “crocin sugar adduct”.

As shown in the following formula (II), the site of crocin (I) to which glucose or a glucose oligomer binds is preferably the 3rd, 4th and 4th of each of the two gentiobiose molecules of crocin represented by the above formula (I). It is at least one place selected from the group consisting of 'positions. The 4 ′ position is preferred.

Here, in the formula (II), Glc means a glucose residue.

Even if there is only one glucose residue that binds to any hydroxyl group of crocin, preferably one selected from the group consisting of the 3rd, 4th and 4'positions of each of the two gentiobiose molecules (For example, in the above formula (II), when at least one of n ₁ to n ₆ is 1), it may be an oligomer of glucose in which two or more glucose residues are linked (for example, the above-mentioned In the formula (II), when at least one of n ₁ to n ₆ is an integer of 2 or more). Here, the linkage mode of the glucose residue in the oligomer is not particularly limited, but is usually an α-1,4 bond or an α-1,6 bond, and preferably an α-1,4 bond.

The number of glucose constituting the glucose oligomer may be 2 or more, and can usually be appropriately selected from the range of 2 to 20. The number is preferably 2 to 10, more preferably 2 to 8, still more preferably 2 to 4, and particularly preferably 2.

More specifically, in the crocin sugar addition compound represented by the above formula (II), the number (n ₁ ) of glucose residues bound to the oxygen atom at the 4 ′ position of the left gentiobiose is in the range of 0-20. , Preferably in the range of 1 to 10, more preferably in the range of 1 to 8, particularly preferably 1 to 5; the number of glucose residues bonded to the oxygen atom at the 4-position (n ₂ ) may include a number selected from the range of 0 to 20, preferably 1 to 10, more preferably 1 to 8, and particularly preferably 1 to 5; The number of glucose residues bonded to the atom (n ₃ ) is selected from the range of 0 to 20, preferably 1 to 10, more preferably 1 to 8, particularly preferably 1 to 5. Numbers can be mentioned. Similarly, in the crocin represented by the above formula (II), the number (n ₄ ) of glucose residues bound to the oxygen atom at the 3-position of the right-side gentiobiose is in the range of 0 to 20, preferably 1 to 10 A number selected from the range of 1 to 8, more preferably 1 to 8, particularly preferably 1 to 5; the number of glucose residues bonded to the oxygen atom at the 4-position (n ₅ ) is: A number selected from the range of 0 to 20, preferably 1 to 10, more preferably 1 to 8, particularly preferably 1 to 5 can be mentioned; bonded to the 4′-position oxygen atom The number of glucose residues (n ₆ ) is a number selected from the range of 0 to 20, preferably 1 to 10, more preferably 1 to 8, particularly preferably 1 to 5. be able to.

In the present invention, a more preferable crocin sugar addition compound and a mixture thereof include a crocin sugar addition compound represented by the formula (II), wherein the number (n ₁ ) of glucose residues bound to the oxygen atom at the 4′-position of one gentiobiose is 1 or more, preferably in the range of 1 to 5, more preferably in the range of 1 to 2, and any number of glucose residues (n ₂ to n ₆ ) bonded to other oxygen atoms 0 compounds, and mixtures thereof. In addition, as another preferable crocin sugar addition compound and a mixture thereof, in the crocin sugar addition compound represented by the formula (II), the number of glucose residues bonded to each 4′-position oxygen atom of two (both) gentiobioses (N ₁ and n ₆ ) are the same or different and each is a number selected from 1 or more, preferably in the range of 1 to 5, more preferably in the range of 1 to 2, and the residual glucose bound to other oxygen atoms Examples thereof include compounds in which the number of groups (n ₂ to n ₅ ) is all 0, and mixtures thereof.

The crocin sugar adduct of the present invention is preferably obtained by allowing glucose residue transferase to act on crocin in the presence of a sugar donor (glucose source) according to a conventional method. As shown, crocin is a crocin glycosylation compound or a mixture thereof glucosylated to various degrees.

The crocin sugar addition compound targeted by the present invention includes structural isomers comprehensively included in the chemical formula (II). The structural isomer is bis [(2S, 3R, 4S, 5S, 6R) -3,4,5-trihydroxy-6-[[(2R, 3R, 4S, 5S, 6R) -3,4,5- trihydroxy-6- (hydroxymethyl) oxan-2-yl] oxymethyl] oxan-2-yl] (2E, 4E, 6E, 8E, 10E, 12E, 14E) -2,6,11,15-tetramethylhexadeca-2,4 , 6,8,10,12,14-heptaenedioate), at least one carbon atom selected from the group consisting of the 3-position, 4-position and 4'-position of each of the two gentiobiose molecules, preferably the 4'-position carbon The crocin sugar addition compound which glucose and a glucose oligomer couple | bond with an atom through an oxygen atom can be mentioned.

As crocin, commercially available crocin may be used, or crocin prepared by extraction and purification from saffron (Crocus sativus L) or gardenia (Yamadenoko, Gardenia jasminoides) as shown in Example 1 described later. May be. Details of the method of extracting and purifying crocin from saffron are described in, for example, Patent Document 2, and can be referred to.

The glucose residue transferase used for glycine glycosylation is not limited, and examples thereof include transglucosidases such as cyclodextrin glucanotransferase (EC2.4.1.19) (hereinafter also referred to as “CGTase”). it can.

These glucose residue transferases are all commercially available enzymes. For example, examples of such commercially available enzyme preparations include contizyme (trade name) (manufactured by Amano Enzyme Co., Ltd.). The amount of glucose residue transferase used as an enzyme is GCTase (enzyme specific activity: about 100 units. One unit means the amount of enzyme that produces 1 mg of β-cyclodextrin per minute from soluble starch. ), For example, the enzyme can be in the range of 1 part by weight to 1 part by weight per 1 part by weight of crocin. The amount is preferably about 1 / 100,000 to 1 / 10,000 parts by weight.

The sugar donor (glucose source) used in the production of crocin sugar adduct is a sugar containing glucose as a constituent sugar, and one or more molecules of glucose residues constituting the sugar are transferred to crocin. If it is. Examples thereof include glucose, maltose, amylose, amylopectin, starch, starch liquefied product (liquefied starch), starch saccharified product (saccharified starch), and cyclodextrin. Here, the starch liquefied product is a product obtained by decomposing starch to the extent that it is solubilized by adding water and an acid or a hydrolase to the starch and reacting it. The starch saccharified product causes the enzyme to act on the starch. In this way, it is broken down into small molecule sugars (small saccharides, monosaccharides).

Preferably, it is starch, starch liquefied product, or starch saccharified product. The amount of the sugar donor (glucose source) used is usually in the range of 1 to 1000 parts by weight, preferably in the range of about 5 to 100 parts by weight, more preferably 10 parts by weight with respect to 1 part by weight of crocin present in the reaction system. A range of about 40 parts by weight can be mentioned.

The crocin sugar adduct varies depending on the enzyme used and is not particularly limited. For example, when GCTase or the like is used as a glucose residue transferase, it is usually at a temperature of about 80 ° C. or lower and at a pH of about 3 to 11. It can be prepared by allowing glucose residue transferase to act on crocin in the presence of the sugar donor (glucose source) under pH conditions. The temperature condition is preferably about 20 to 80 ° C. or less, more preferably about 40 to 75 ° C. The pH condition is preferably pH 4 to 8. The reaction time is not particularly limited, but is usually 0.1 hour or more, preferably 1 to 24 hours, more preferably 3 to 6 hours.

Such a reaction can be performed while standing or stirring or shaking. In order to prevent oxidation during the reaction, the head space of the reaction system may be replaced with an inert gas such as nitrogen.

Thus, a composition containing a mixture of crocin sugar addition compounds (II) including a plurality of types of crocin sugar addition compounds glucosylated to various degrees is obtained. The composition contains raw materials and by-products such as unreacted crocin, sugar donor (glucose source), and glucose residue transferase. For this reason, you may perform the process which removes these raw materials and a by-product as needed.

For such treatment, conventional purification methods can be carried out singly or in various combinations, for example, extraction methods, solvent fractionation methods, salting out methods, acid precipitation methods, recrystallization methods, activated carbon treatment methods, electrodialysis methods, various types of methods. Resin treatment methods (adsorption method, ion exchange treatment method, column chromatography such as gel filtration method), and membrane treatment methods (ultrafiltration membrane treatment method, reverse osmosis membrane treatment method, ion exchange membrane treatment method, Zeda potential Examples thereof include a film processing method.

In addition, it is possible to isolate and acquire individual crocin sugar addition compounds from the mixture by subjecting the thus obtained mixture of crocin sugar addition compounds to separation means such as high performance liquid chromatography.

(B) Use of crocin sugar addition compound or a mixture thereof As shown in Experimental Example 2 described later, the crocin sugar addition compound of the present invention or a mixture thereof (crocin sugar addition product) is compared with crocin in the body by oral administration. It has excellent absorbability (transferability in blood) (oral absorbability), and as a result, it has the feature of exhibiting an excellent sleep improvement effect or sedation effect at a dose smaller than that of crocin.

Here, “improving sleep” generally means inducing a good sleep state by inducing a good sleep by actions such as smoothing sleep, reducing arousal during sleep, and improving sleepiness when waking up. To do. As shown in Experimental Example 2, the crocin sugar adduct of the present invention has an action of prolonging non-REM sleep, which is an indicator of deep sleep. Therefore, by taking the crocin sugar adduct of the present invention orally, the quality of sleep is improved, and poor physical condition and discomfort caused by shallow sleep, such as mid-wake awakening, discomfort and sleepiness when waking up The effect of eliminating or improving can be exhibited. That is, “sleep improvement” as used in the present invention preferably means improvement of sleep based on the action of prolonging non-REM sleep possessed by a crocin sugar adduct.

Also, the crocin sugar adduct of the present invention has an action (sedation action) of reducing active behavior as shown in Experimental Example 3 described later.

The crocin sugar adduct of the present invention is superior in stability in aqueous solution and / or stability to light irradiation as compared to crocin as shown in Experimental Example 1 described later. For this reason, it is suitable as a component to be blended in a product distributed on the market, and can be effectively used particularly for the production of a product aimed at improving sleep and / or sedation.

The composition containing the crocin sugar adduct targeted by the present invention can be used as a pharmaceutical, a quasi-drug, a food or drink, and the like.

As a medicine or quasi-drug, it contains crocin sugar adduct as an active ingredient, sleep improvement effect as sleep improvement agent etc. (sleep promotion, sleep, shorten sleep time, dose of sleep agent used in combination as necessary Can be mentioned that promoted (reduction). Moreover, what contains crocin sugar adduct as an active ingredient and appealed the sedative effect as a sedative can be mentioned.

The dosage form of the pharmaceutical or quasi-drug may be either oral or parenteral. The oral dosage forms include powders, granules, capsules (hard capsules, soft capsules), tablets, pills, Solid agents such as chewables and lozenges; and liquids such as solutions and syrups; and parenteral agents include injections, sprays (such as nasal drops), suppositories, and the like. As described above, the preferred dosage form is oral administration because the active ingredient crocin sugar adduct is excellent in oral absorbability.

Such a pharmaceutical product or quasi-drug may be composed only of the crocin sugar addition compound represented by the above formula (II) or a mixture thereof, but depending on the use form such as the above administration form, a conventional carrier It may contain other components such as a base material or an additive, and can be formulated using a known formulation method, particularly a formulation technique suitable for oral intake. For example, a pharmaceutical product or quasi-drug can be produced by mixing an effective amount of the above crocin sugar adduct as an active ingredient with a pharmaceutically acceptable carrier or additive.

Such a carrier can be selected according to a conventional method according to the administration form. For example, when the dosage form has a solid form such as powders, granules, pills, capsules, chewables, troches and tablets, excipients such as lactose, glucose, sucrose, mannitol; starch, sodium alginate, etc. Disintegrating agents; lubricants such as magnesium stearate and talc; binders such as polyvinyl alcohol, hydroxypropyl cellulose and gelatin; surfactants such as fatty acid esters; plasticizers such as glycerin and the like. In the case of liquid forms such as solutions, suspensions and syrups, water; sugars such as sucrose, sorbitol and fructose; glycols such as polyethylene glycol and propylene glycol; oils such as sesame oil, olive oil and soybean oil Etc. can be used. Moreover, as an additive, auxiliary agents, such as a chelating agent, antiseptic | preservative, a fragrance | flavor, a corrigent, etc. can be used, for example.

The proportion of crocin sugar adducts contained in drugs or quasi-drugs varies depending on the form of use, intended effect and extent, gender and age of the user, etc. The ratio of the total amount of the crocin sugar addition compound represented by the formula (II) or the mixture thereof is 0.01 to 100% by weight, preferably 0.1 to 10% by weight, based on the total weight of the product.

When a drug or quasi-drug is administered to a subject, preferably a subject (patient) complaining of insomnia or a subject who wants good quality sleep (patient), the effective dose is determined by the age, weight, and pathological condition (sleep) of the subject patient. The degree of disorder) and the administration method, etc., but generally the total amount of the active ingredient crocin sugar adduct is about 0.001 to 100 mg / kg / day, preferably about 2.5 to 20 mg / kg / day. To be administered. Moreover, when administering to the test subject who desires sedation, the ratio similar to the above can be mix | blended. The administration period is usually from immediately before going to bed to 3 hours before going to bed, more preferably from just before going to bed to 1 hour before going to bed.

In addition, as food and drink containing the crocin sugar adduct of the present invention, the crocin sugar adduct itself of the present invention, or a preparation prepared by adding the aforementioned carrier or additive to the crocin sugar adduct (for example, For example, supplements such as powders, granules, tablets, capsules (hard capsules, soft capsules), liquids, drinks, etc. can be mentioned, and crocin sugar addition of the present invention is added to general foods and drinks. Functional foods (including special health foods and conditional special health foods) that are added to food and drinks and have functions for living organisms (for example, sleep-improving functions such as sleep promotion and sleep) Can be mentioned. These functional foods contain the above-mentioned crocin sugar adduct of the present invention, and are characterized by having a sleep improving action (for example, including a sleep promoting action or a resting action) or a sedative action. In addition, a food or drink with a display indicating that it is used for improving sleep or that it is used for calming is included.

The ratio of the crocin sugar adduct added to the food or drink is not particularly limited as long as it has the function, but can be appropriately selected from the range of 0.001 to 100% by weight. The target food and drink is not limited, but ice cream, ice milk, lacto ice, sherbet, ice confectionery and other frozen confectionery; milk beverage, lactic acid bacteria beverage, soft drink (including fruit juice), carbonated drink, fruit juice drink, Beverages such as vegetable beverages, vegetable / fruit beverages, sports beverages, powdered beverages; alcoholic beverages such as liqueurs; tea beverages such as coffee beverages and tea beverages; soups such as consomme soup and potage soup; custard pudding, milk pudding Puddings such as pudding with fruit juice, desserts such as jelly, bavaroa and yogurt; gums such as chewing gum and bubble gum (plate gum, sugar-coated granule gum); coated chocolate such as marble chocolate, strawberry chocolate, blueberry Chi with flavors such as chocolate and melon chocolate Chocolate such as collate; hard candy (including bonbon, butterball, marble, etc.), soft candy (including caramel, nougat, gummy candy, marshmallow, etc.), caramel such as drop, toffee, etc .; hard biscuits, cookies, rice cake Baked confectionery such as rice crackers; fruit wine such as red wine; and other various processed foods and drinks such as various side dishes.

In addition, the intake of the food and drink of the present invention is not particularly limited as long as it is ingested and exhibits a sleep improving effect or a sedative effect in the body. For example, in the case of an adult with a body weight of 60 kg, The sugar adduct can be appropriately selected from a range including a total amount of 1 mg to 30 g.

(C) Method for Increasing Oral Absorption of Crocin The present invention provides a method for enhancing the oral absorption of crocin. According to the method of the present invention, it is possible to increase the oral absorbability of crocin or crocetin, which is an aglycon of crocin known to have a sleep-improving or sedative effect, and as a result, sleep of crocin or crocetin in vivo. It becomes possible to enhance the improving action or the sedative action.

The method of the present invention specifically uses crocin represented by the formula (I) as a raw material:

This can be carried out through a step of producing a crocin sugar adduct compound represented by the following formula (II) by allowing a glucose residue transferase to act in the presence of a sugar donor.

The definitions of crocin sugar addition compound, sugar donor and glucose residue transferase and the examples thereof are as described above in (A) and can be incorporated herein. The conditions for the above action are also as described in (A), and can be used here.

In this method, a conventional purification method can be arbitrarily combined and thus a crocin sugar addition compound or a mixture thereof with higher purity can be obtained.

According to the method of the present invention, when orally administered, crocin can be converted into a crocin sugar addition compound or a mixture thereof that exhibits 10-fold or more absorption in the body compared to crocin. Preferably it is 100 times or more, More preferably, it is 200 times or more.

Hereinafter, examples and experimental examples will be described in order to show the configuration and effects of the present invention in more detail. However, the present invention is not limited to these examples.

Example 1 Production method of crocin sugar adduct (1) Preparation of crocin A filtrate (extract) obtained by extracting dry saffron with 95% ethanol at room temperature and solid-liquid separation was dried under reduced pressure. This was redissolved in water and then liquid-liquid partitioned with dichloromethane to remove insoluble components. Next, the aqueous phase was roughly purified by chromatography and then subjected to high performance liquid chromatography (HPLC) to obtain crocin.

(2) Addition of sugar to crocin The crocin prepared by the above method was dissolved in a 10% (w / v) maltosyl-β-cyclodextrin aqueous solution to prepare a 5 mg / mL solution. Next, this was mixed with a 10% (w / v) soluble starch (starch liquefied) solution (manufactured by Nacalai Tesque) and crocin 2.5 mg / mL, 5% (w / v) maltosyl-β-cyclodextrin And 5% (w / v) starch-containing aqueous solution was prepared, and cyclodextrin glucatransferase (CGTase) (manufactured by Amano Enzyme) was added thereto and mixed so as to have a final concentration of 5 U / ml. CGTase is an enzyme having a catalytic action of starch hydrolysis reaction and cyclodextrin synthesis reaction, as well as an action of catalyzing a sugar transfer reaction between different sugar molecules (Non-patent Document 2). For this reason, by using CGTase, as shown in the following formula, glucose or an oligomer of glucose produced by hydrolysis of starch can be added to gentiobiose of crocin.

The prepared mixed solution was allowed to stand at 40 ° C. for 1 to 6 hours under light shielding to carry out a sugar transfer reaction to crocin.

1 hour, 1 hour, 3 hours and 6 hours after the reaction, the reaction solution was collected and subjected to HPLC under the following conditions, and the result of confirming the production status of the reaction product is shown in FIG.

<HPLC conditions>
HPLC system: SHIMADZU Prominence LC (LC-20A)
Column: COSMOSIL Cholester 4.6 mm ID × 150 mm
Mobile phase A: 10 mM ammonium acetate aqueous solution Mobile phase B: 10 mM ammonium acetate methanol solution flow rate: 0.6 mL / min
Detection: UV-Vis detection 440 nM.

As can be seen from FIG. 1, it was confirmed that sugar was added to crocin in a time-dependent manner in the reaction with CGTase. The glycosylation rate after 6 hours from the start of the reaction was 93%. When 1,4-α-D-Glucan glucohydrolase (glucoamylase) was allowed to act on this crocin sugar adduct, sugar (glucose, oligomer of glucose) was dissociated and crocin was detected. In addition, the added saccharide could be completely eliminated by glucoamylase treatment without affecting gentiobiose (1-6 bond). From these facts, it is confirmed that the sugar chain (glucose residue) is not a 1 → 6 bond but an α- (1 → 4) bond, and no sugar chain is bonded at least at the 6 ′ position. (See Experimental Example 4).

Based on the above results, by acting CGTase in the presence of crocin and starch, the hydroxyl group of gentine biose of crocin (3rd, 4th and / or 4 'position of crocin's gentiobiose molecule, preferably 4' position) It is considered that glucose (Glc) produced by hydrolysis of starch or an oligomer of glucose is condensed to produce a crocin sugar adduct of the following formula (II).

Experimental Example 1 Stability evaluation of crocin sugar adduct Since crocin or crocetin has a conjugated double bond in the molecule, it is susceptible to auto-oxidation and easily decomposes. In addition, crocin has a problem that glucose on both sides is easily cleaved by saffron endogenous β-glucosidase.

Therefore, in this Experimental Example 1, the stability (stability in aqueous solution, light stability) of the crocin sugar adduct produced in Example 1 was examined and compared with that of crocin.

(1) Experimental method (1-1) Stability evaluation in aqueous solution The crocin produced in Example 1 was dissolved in 5% (w / v) maltosyl-β-cyclodextrin aqueous solution, and the enzyme ( After treatment with CGTase), the enzyme was inactivated by heat treatment to prepare a crocin sugar adduct-containing solution containing unreacted crocin (n = 0). This was allowed to stand for 6 hours at room temperature under light-shielding conditions. At 0 hours, 1 hour, 3 hours and 6 hours after standing, a part of the reaction solution was collected and subjected to HPLC, and the contents of crocin sugar adduct and crocin in the aqueous solution were evaluated and confirmed from the absorbance.

(1-2) Stability evaluation against light irradiation After the crocin produced in Example 1 was dissolved in 5% (w / v) maltosyl-β-cyclodextrin aqueous solution and treated with an enzyme (CGTase) in the presence of starch, The enzyme was inactivated by heat treatment to prepare a crocin sugar adduct-containing solution containing unreacted crocin (n = 0). This was allowed to stand for 6 hours at room temperature and under light irradiation conditions (fluorescent lamp irradiation of about 1000 lux). After 0 hours, 1 hour, 3 hours and 6 hours from light irradiation, a part of the reaction solution was collected and subjected to HPLC, and the contents of crocin sugar adduct and crocin were evaluated and confirmed from the absorbance.

Regarding stability in aqueous solution (room temperature), as shown in FIG. 2 (A) and Table 1, crocin itself decomposes 25% 6 hours after preparation in aqueous solution and 48% after 24 hours. In contrast, all of the crocin sugar adducts were found to remain 87% or more after 6 hours and 67% or more after 24 hours without being degraded.

As for the stability against light irradiation, as shown in FIG. 2 (B) and Table 2, crocin decomposed 32% after 6 hours from light irradiation and 60% after 24 hours. It was found that all sugar adducts remained without being decomposed by 84% or more even after 6 hours and 55% or more after 24 hours.

From the above results, crocin is an unstable compound in an aqueous solution (room temperature) and under light irradiation conditions, whereas the crocin sugar adduct of the present invention in which sugar (glucose or glucose oligomer) is added to crocin. It was also found that the stability of the aqueous solution was improved and it was more stable in aqueous solution and light irradiation than crocin.

Experimental Example 2 Evaluation of the efficacy of crocin sugar adduct (Part 1)
About the crocin sugar adduct manufactured in the said Example 1, the influence which it has on sleep and a spontaneous exercise amount was investigated using the mouse | mouth, and it compared with the influence which crocin has on sleep. For administration to mice, an aqueous solution of crocin sugar adduct (2.5 mg / ml) prepared by dissolving in physiological saline was used as the crocin sugar adduct. Also, crocin was used as a crocin suspension (0.8 mg / ml) prepared by suspending in physiological saline. For control, a 5% (w / v) maltosyl-β-cyclodextrin aqueous solution was used as a solvent to be administered to mice.

(1) Experimental method (1-1) Animals used C57BL / 6 male mice (10-12 weeks old, SLC Japan) were used for sleep measurement experiments. Each mouse had free access to radiation-sterilized food (lab MR stock) and water at a temperature of 22 ° C. (± 2 ° C.), a humidity of 55% (± 5%), and a 12 hour light-dark cycle.

(1-2) Sleep measurement experiment Somnopentyl (Kyoritsu Pharmaceutical Co., Ltd., general name: pentobarbital, 50 mg / kg, intraperitoneal administration) Under anesthesia, an operation was performed in which an electrode for measuring an electroencephalogram / myoelectric potential was implanted in a mouse. The electrode was connected to a microconnector and fixed to the skull of the mouse with dental cement. After a 10-day recovery period from the surgery, the mice were transferred to another recording cage, connected to electrodes and cables fixed to the skull, and further acclimated for 3 days. A split ring was installed between the cable and the head amplifier so that the behavior of the mouse was not restricted.

After the acclimatization period, the electroencephalogram and myoelectric potential were recorded for 3 days from the beginning of the dark period (starting at 20:00) in a 12 hour light / dark cycle (light period: 7: 00-19: 00). The first day was the base measurement day, the second day was the test day, and the third day was the recovery measurement day. Specifically, on the first day of base measurement, the electroencephalogram and myoelectric potential were measured after administering a solvent (5% (w / v) / maltosyl-β-cyclodextrin aqueous solution) (control), then 2 days. Immediately before the start of the dark period (20:00) on the eye test day, crocin sugar adduct aqueous solution (2.5 mg / kg or 25 mg / kg) was orally administered. As a comparative example, crocin suspension (10 mg / kg or 30 mg / kg) was intraperitoneally administered. Next, the electroencephalogram and myoelectric potential were measured, and the effects of crocin and crocin sugar adduct on sleep / wakefulness were examined. Calculate the cumulative amount for a certain period of time (4 hours, 6 hours) after administration, and compare with the cumulative amount in the test group (crocin sugar addition group), comparison group (crocin administration group), and control group (solvent administration group) did.

The brain wave and myoelectric potential were recorded after being amplified and filtered (brain wave: 0.5-300.5Hz, myoelectric potential: 20-200 Hz), analog-digital converted at a sampling rate of 128 Hz. The electroencephalogram analysis uses software SleepSign (registered trademark) (manufactured by Kissei Comtech) as 10 epoch data, and each epoch is awakened, non-REM sleep, or REM sleep by the frequency component and waveform of the electroencephalogram and myoelectric potential. It was determined to be either (Non-Patent Document 3). Here, the “epoch” refers to an arbitrary time section for performing sleep / wake stage determination. The specific determination method is to first determine that the infrared sensor has responded to arousal, and by frequency analysis, those that contain a particularly large amount of delta wave (0.65-4.0 Hz) components are non-REM sleep and theta waves (6.0-10.0). Hz) A component having a high component and a low myoelectric potential was determined as REM sleep. The others were determined to be awake.

(1-3) Bioavailability evaluation experiment When crocin is orally administered, crocin is hydrolyzed and absorbed in the intestine and detected as crocetin in the blood. Therefore, crocin sugar adducts were intraperitoneally or orally administered to mice, and blood was collected 15 minutes, 30 minutes and 60 minutes after administration, and the amounts of crocin sugar adduct, crocin and crocetin in the blood were collected. Was measured.

(2) Experimental results (2-1) Sleep measurement results For the test group, the cumulative sleep amount for 6 hours after oral administration of crocin sugar adduct (25 mg / kg) was calculated for each awakening time, REM sleep time, and non-REM sleep time. The results compared with each of the control group (solvent administration group) are shown in FIG. In addition, the cumulative sleep amount (REM sleep time and non-REM sleep time) for 4 hours after oral administration of crocin sugar adduct (2.5 mg / kg, 25 mg / kg) for the test group and intraperitoneal crocin suspension for the comparison group FIG. 3 (B) shows the result of comparison with the cumulative amount of sleep (REM sleep time and non-REM sleep time) for 4 hours after administration (10 mg / kg, 30 mg / kg).

As shown in FIGS. 3 (A) and 3 (B), according to the crocin sugar adduct, it was confirmed that a sleep enhancing effect was obtained at an oral dose of 25 mg / kg. As shown in FIG. 3 (B), in order to obtain a sleep promoting effect with crocin, it was necessary to administer 30 mg / kg or more of crocin intraperitoneally, but in the case of the crocin sugar adduct of the present invention, Since the same effect can be obtained by oral administration at 25 mg / kg, it is considered that the action effect (sleep promoting effect) of crocin in the body is enhanced by using crocin as a sugar adduct.

(2-2) Results of bioavailability evaluation The results are shown in FIG.

As shown in FIG. 4 (A), when the crocin sugar adduct was administered intraperitoneally, only the crocin sugar adduct and crocin were detected 15 to 60 minutes after administration, and crocetin was not detected. On the other hand, as shown in FIG. 4 (B), when the crocin sugar adduct was orally administered, crocetin (7.67 μM) was detected in the blood 15 minutes after the administration. The blood crocetin concentration reached its maximum (22.3 μM) 30 minutes after administration, and 12.2 μM crocetin was detected even 60 minutes later.

From the results shown in Experimental Examples 1 and 2 above, it was found that the following effects can be obtained by adding sugar to crocin using enzymatic glycosyl transfer.
(1) Stability in aqueous solution and stability against light irradiation are improved compared to crocin (Experimental Example 1)
(2) Oral absorbability is improved rather than using crocin itself, and a sleep improvement effect is demonstrated by oral administration at a low dose (Experimental Example 2).
thing.

That is, the crocin sugar adduct of the present invention is excellent in stability in an aqueous solution or under light irradiation conditions, and is excellent in sleep improvement effect. It can be used effectively for the production of food and drink.

Experimental Example 3 Evaluation of drug efficacy of crocin sugar adduct (2)
1. experimental method
(i) Animals For behavioral measurements (spontaneous exercise measurement experiments), 12-week-old C57BL / 6 male mice, body weight 25-30 g, SLC Japan), 12-week-old adenosine A1 receptor gene-deficient mice ( C57BL / 6 male mice weighing 25-30 g) and 12-week old histamine H1 receptor gene-deficient mice (C57BL / 6 male mice weighing 25-30 g) were used. Mice were given free access to radiation-sterilized diet (lab MR stock) and water at a temperature of 22 ± 2 ° C., humidity of 55 ± 5%, and a 12-hour light-dark cycle.

(Ii) Activity measurement and analysis To measure activity, mice were acclimated to the environment in an animal activity measurement chamber for 2 days, and then the solvent (5% (w / v) maltosyl-β-cyclodextrin Aqueous solution) was orally administered. Immediately after administration, mice were placed in individual cages and behavioral measurement was started. Twenty-four hours after the administration of the solvent, the crocin sugar adduct was orally administered, and then the mice were placed in individual cages to measure the amount of behavior. The amount of behavior was recorded using a sensor that detects infrared rays emitted from the animal and the software Biotex 16CH Act Monitor BAI2216 (Biotex Japan). The behavioral amount was calculated by calculating the cumulative amount every hour after administration of crocin sugar adduct and the cumulative amount after 12 hours, and comparing it with the behavioral amount group after solvent administration.

2. Experimental results 1) Effects on the amount of activity of C57BL / 6 mice As shown in the top of Fig. 5, the crocin sugar adduct (2.5 mg / kg) was applied to the C57BL / 6 mice in the dark phase (19:00) When administered orally immediately before the administration, the amount of behavior was statistically significantly suppressed from the administration of the solvent until 2 hours after the administration. In addition, when the cumulative behavioral amount 12 hours after administration of the crocin sugar adduct was compared with the solvent administration, a statistically significant suppression of behavioral amount was observed.

2) Effects on the behavioral amount of adenosine A1 receptor gene-deficient mice As shown in the middle part of Fig. 5, acrosin sugar adduct (2.5 mg / kg) was started in the dark period in adenosine A1 receptor gene-deficient mice (19:00) ), The amount of behavior was statistically significantly suppressed from 1 hour to 4 hours after administration, compared with the solvent administration. In addition, when the cumulative behavioral amount 12 hours after administration of the crocin sugar adduct was compared with the solvent administration, a statistically significant suppression of behavioral amount was observed.

3) Effect on the amount of behavior of histamine H1 receptor gene-deficient mice As shown in the bottom of Fig. 5, crocin sugar adduct (2.5 mg / kg) was started in the dark period in histamine H1 receptor gene-deficient mice (19: No difference was observed in the amount of behavior after oral administration immediately before 00) compared to the solvent administration. In addition, there was no difference in the cumulative behavioral amount for 12 hours after the administration of crocin sugar adduct as compared with the solvent administration.

From the above results, it was confirmed that the amount of action in the active period (dark period) was suppressed by the administration of crocin sugar adduct (behavior suppressing effect). Moreover, since the behavioral inhibitory effect of crocin sugar adduct is attenuated in histamine H1 receptor gene-deficient mice, the behavioral inhibitory effect (sedation effect) by oral administration of crocin sugar adduct is mediated by histamine receptor It is considered to be an action.

Experimental Example 4 Identification of crocin sugar adduct The reaction solution 6 hours after the reaction in Example 1 was subjected to HPLC, and each peak was separated from each peak (1, 2, 3, 4 and 5 in order from the right in FIG. 1). Each fraction (Fraction-1, 2, 3, 4 and 5) taken was subjected to mass spectrum and / or NMR spectrum (1H NMR and 13C NMR) to identify the reaction product.

Fig. 6 shows the results of mass spectrum. In order from the top, mass spectra of Fraction-1, 2, 3, 4 and 5 are shown. The measurement conditions for the mass spectrum are as follows.

<Measurement conditions of mass spectrum>
Mass spectrum measuring apparatus: API 3200 manufactured by AB SCIEX
Ionization conditions: Electron sprayionization (ESI), Negative mode
Mobile phase: 40% MeOH, 10 mM ammonium acetate Scanning range: m / z 800-1700.

From this result, the compound of Fraction-1 (peak 1) is crocin (molecular weight: 975.7 (MH) ⁻ ); the compound of Fraction-2 (peak 2) is a crocin sugar addition compound (crocin) obtained by dehydration condensation of one molecule of glucose to crocin. mono-glycoside) (molecular weight: 1137.6 (MH) ⁻ ); the compound of Fraction-3 (peak 3) is a crocin di-glycoside compound obtained by dehydration condensation of two glucose molecules on crocin (molecular weight: 1299.4 (MH) ^- ); Fraction-4 (Peak 4) is a crocin tri-glycoside compound obtained by dehydration condensation of 3 molecules of glucose to crocin (molecular weight: 1461.4 (MH) ^- ); Fraction-5 (Peak 5) The compound was estimated to be crocin tetra-glycoside (molecular weight: 1623.6 (MH) ⁻ ) obtained by dehydration condensation of 4 molecules of glucose to crocin.

Fraction-1, 2, and 3 were subjected to ¹³ C-NMR spectrum (solvent: dimethyl sulfoxide-d6 [DMSO-d6]) (13C-NMR data). FIG. 7 shows ¹ H-NMR spectrum (solvent : DMSO-d6) (1H-NMR data) is shown in FIG. The NMR measurement conditions are as follows.

<NMR measurement conditions>
NMR measuring apparatus: ECA800 manufactured by JEOL
¹ H spectrum measured at 800 MHz, ¹ H
^{The 13} C spectrum was measured at 200 MHz and ¹³ C. Solvents: DMSO-d6, D ₂ O (both 99.6% purity, manufactured by ISOTEC).

From this result, the compound of Fraction-1 (peak 1) is crocin, and the compound of Fraction-2 (peak 2) has one glucose residue at the 4'-position oxygen atom of one gentiobiose of crocin. 1 → 4) It is a crocin mono-glycoside bonded compound, and the compound of Fraction-3 (Peak 3) is composed of two molecules of glucose on the oxygen atom at the 4 'position of gentiobiose on one side of crocin. The α- (1 → 4) conjugate was found to be an α- (1 → 4) linked crocin sugar di-glycoside (FIG. 9).

Furthermore, this indicates that when these crocin sugar adducts are digested with glucoamylase (1,4-α-D-Glucan glucohydrolase), the crocin gentiobiose (6-O-β-D-glucopyranosyl-D-glucose (6- It was also confirmed from the fact that only crocin was obtained without affecting o-βD-Glycopyranosyl-D-glucose).

Formulation Example 1: Tablet A tablet (oral drug or food) is prepared according to the following composition by a conventional method.
Crocin sugar adduct (Example 1) 200 mg
Lactose 60mg
Potato starch 30mg
Polyvinyl alcohol 2mg
Magnesium stearate 1mg
Tar pigment trace amount.

Formulation example 2: Powder A powder (oral drug or food) is prepared according to the following composition according to a conventional method.
Crocin sugar adduct (Example 1) 200 mg
Lactose 275 mg.

Claims

A crocin sugar addition compound or a mixture thereof, wherein glucose or a glucose oligomer is condensed on one or more hydroxyl groups of crocin (I) represented by the following formula:
The crocin sugar addition compound or mixture thereof according to claim 1, which is represented by the following formula (II):

(In the formula, Glc is a glucose residue, and n 1 to n 6 are the same or different and each represents an integer of 0 to 20, except that all of n 1 to n 6 are 0.) .
The crocin sugar addition compound according to claim 2 is a compound in which n 1 is 1 or 2 and n 2 to n 6 are 0 in formula (II), and the mixture according to claim 2 is the crocin The crocin sugar addition compound or mixture thereof according to claim 2, comprising a sugar addition compound.
The crocin sugar addition compound or a mixture thereof according to any one of claims 1 to 3, wherein crocin (I) is produced by a method in which a glucose residue transferase is allowed to act in the presence of a sugar donor.
The crocin sugar adduct compound or a mixture thereof according to claim 4, wherein the sugar donor is a glucose source.
Crosin (I) represented by the following formula:

A method for producing a crocin sugar adduct compound represented by the following formula (II) or a mixture thereof, which comprises a step of allowing glucose transferase to act in the presence of a sugar donor

(In the formula, Glc is a glucose residue, and n 1 to n 6 are the same or different and each represents an integer of 0 to 20, except that all of n 1 to n 6 are 0.) .
The crocin sugar addition compound according to claim 6 is a compound in which n 1 is 1 or 2 and n 2 to n 6 are 0 in formula (II), and the mixture according to claim 6 is the crocin The production method according to claim 6, comprising a sugar addition compound.
The production method according to claim 6 or 7, wherein the sugar donor is a glucose source.
A composition containing the crocin sugar addition compound according to any one of claims 1 to 5 or a mixture thereof.
The composition according to claim 9, which is an oral composition.
The composition according to claim 9 or 10, which is a pharmaceutical, a quasi-drug, or a food or drink.
The composition according to any one of claims 9 to 11, which is a sleep improving agent or a sedative.
A method for improving sleep of an animal, comprising a step of administering the crocin sugar adduct compound or the mixture thereof according to any one of claims 1 to 5 to the animal having a sleep disorder.
A method for sedation of an animal, comprising a step of administering the crocin sugar addition compound according to any one of claims 1 to 5 or a mixture thereof to the animal to be sedated.
Formula (I):

In the presence of a sugar donor, glucose residue transfer enzyme is allowed to act on crocin represented by formula (II):

A method for enhancing the oral absorbability of crocin, comprising the step of producing a crocin sugar adduct compound represented by