WO2001093881A1

WO2001093881A1 - Immunological substance containing enteroabsorbable anti-inflammatory factor and application thereof

Info

Publication number: WO2001093881A1
Application number: PCT/JP2001/004516
Authority: WO
Inventors: Naoyuki Kamata
Original assignee: Ortho Corporation Co., Ltd.; Nomoto, Kikuo
Priority date: 2000-06-02
Filing date: 2001-05-30
Publication date: 2001-12-13
Also published as: AU2001262672A1; JP2001342127A

Abstract

An immunological substance containing an enteric anti-inflammatory factor which is absorbed in the intestine and fully exerts its effect without any deterioration in the effects of the anti-inflammatory factor containing immunological substance in the stomach; and health foods to which the above substance is applied. Namely, an anti-inflammatory factor containing immunological substance coated or matrixed with a substance hardly soluble in the stomach or a substance slowly soluble in the gastric juice. The anti-inflammatory factor containing immunological substance may be directly coated or matrixed with the substance hardly soluble in the stomach or the substance slowly soluble in the gastric juice. Alternatively, the surface of capsules or microcapsules having the anti-inflammatory factor containing immunological substance enclosed therein may be coated or matrixed with the substance hardly soluble in the stomach or the substance slowly soluble in the gastric juice.

Description

Description Immune substance containing intestinal absorptive anti-inflammatory factor and its application

The present invention relates to an intestinal-absorbing anti-inflammatory factor-containing immunological substance and its application. In particular, it is intended to increase the absorption in the intestine of an immunological substance containing an anti-inflammatory factor, which is weak against heat, acid or organic solvent, without impairing its effect in the stomach as much as possible. Background art

Inflammation is "a local reaction that, when damaged or destroyed, causes the tissue and destroys, dilutes or isolates both the injured tissue" and "inflammation, pain, fever, redness, swelling, and function." It is accompanied by signs such as “loss” (Kichiichi Land Medical Encyclopedia). When inflammation occurs, anti-inflammatory drugs are generally given. As anti-inflammatory drugs, antihistamines, steroidal and non-steroidal anti-inflammatory drugs, anti-inflammatory enzyme agents, immunosuppressants and the like are known.

However, each of these anti-inflammatory drugs has the problem of causing side effects and adverse reactions. For example, salicylates, one of the most common anti-inflammatory drugs, aspirin, are well known, but at high doses can cause central respiratory paralysis and circulatory collapse, epigastric pain, nausea, and vomiting And gastrointestinal bleeding.

From this point of view, the emergence of anti-inflammatory agents that do not cause side effects or adverse reactions is desired. An anti-inflammatory drug composition derived from milk has been proposed as an anti-inflammatory agent having no such side effects or adverse reactions (Japanese Patent Application Laid-Open Nos. 57-1888523, 59-1616-131616). ) Further, the isolation of the anti-inflammatory factor and its use as an anti-inflammatory drug have been proposed (Japanese Patent Application Laid-Open Nos. 2-503802, 8-502005 15 Hei 8-502718, Tokuhoku Hei 10-509420).

An object of the present invention is to provide a technique for utilizing an anti-inflammatory factor-containing immunological substance including an anti-inflammatory factor derived from milk as a so-called health food. Here, the anti-inflammatory factor-containing immunological substance refers to attenuated or killed microorganisms, an attenuated virus or an inactivated virus, or the milk of a female mammal that has been hyperimmunized with an adjuvant-mixed vaccine using toxoid. Is an immunogenic substance obtained by mixing a milk protein fraction and a low molecular weight fraction having a molecular weight of 10,000 or less, separated from whey obtained after defatting and decasing using as a raw material. The milk protein fraction and the fractions of the low molecular weight fraction having a molecular weight of 10,000 or less can be obtained from whey by, for example, separation by chromatography, separation by gel filtration, or separation by ultrafiltration. Alternatively, the methods described in JP-T2-503802, JP-T 8-502515, JP-T 8-502718. JP-T 10-509420 can also be used. The substance has an intestinal environment-improving effect and concomitant immunopotentiating and anti-inflammatory effects on humans and animals. The substance can be injected orally or anus into the body in the form of a solution, powder, granules or a form added to food or drink. Specific effects include improving the intestinal environment, improving constipation, preventing and treating colon cancer, preventing and treating opportunistic infections, preventing and treating neoplastic diseases, alleviating immune disorders, and preventing periodontal disease. Treatment, prevention and treatment of gingivitis, prevention and treatment of ulcer disease, relief of rheumatic symptoms, prevention and treatment of asthma.

As described above, an anti-inflammatory factor-containing immunological substance is a useful substance that is effective without causing side effects on anti-inflammatory and immunity. This anti-inflammatory effect can be confirmed, for example, by an air porch method. The principle of the air porch method is to orally administer a substance with an anti-inflammatory effect, create an air porch (air cavity) under the mouse, inject carrageenan (a source of inflammation), and measure the number of leukocytes migrating. Is to do. At this time, if inflammation develops, leukocytes migrate to the affected area and are infiltrated, so the greater the number of leukocytes, the greater the degree of inflammation. Conversely, the stronger the anti-inflammatory effect, the smaller the number of leukocytes that migrate and infiltrate.

Since immunological substances containing anti-inflammatory factors are weak to acids, their effects are impaired in the stomach, and their effects are small when absorbed in the intestine. The present invention enhances intestinal absorption of an anti-inflammatory factor-containing immunological substance without significantly impairing its effect in the stomach. is there. At the same time, we will seek to apply this intestinal absorbable anti-inflammatory factor-containing immunological substance. Disclosure of the invention

The present invention is an intestinal-absorbable anti-inflammatory factor-containing immunological substance in which an anti-inflammatory factor-containing immunological substance is matrixed or coated with a substance that is poorly soluble in gastric juice or a substance that gradually dissolves in gastric juice. The shape can be in the form of powder, granules, tablets or chews.

Further, the present invention is characterized in that a powdery or granular anti-inflammatory factor-containing immunological substance coated or made into a matrix with a substance that is hardly soluble in gastric juice or a substance that gradually dissolves in gastric juice is encapsulated in a capsule. It is an intestinal absorbable anti-inflammatory factor-containing immunological substance.

And a matrix or coating of an anti-inflammatory factor-containing immunological substance with a poorly soluble gastric fluid or a substance that gradually dissolves in gastric juice, or a coating or matrix with a poorly soluble gastric fluid or a substance that gradually dissolves in gastric juice. Including powdered or granular anti-inflammatory factor-containing immunological substance encapsulated in a capsule, the thickness of the coated gastric fluid poorly soluble substance or the substance that slowly dissolves in gastric juice, It is possible to change the proportion of substance components that gradually dissolve in gastric juice.

Furthermore, the present invention relates to a method for preparing a powdery or granular anti-inflammatory factor-containing An intestinal absorptive anti-inflammatory factor-containing immunological substance with a sealed substance, containing a powdery or granular anti-inflammatory factor in a microcapsule coated on its surface with a poorly soluble substance in gastric juice or a substance that slowly dissolves in gastric juice. It is an intestinal absorptive anti-inflammatory factor-containing immunological substance in which the immune substance is sealed.

Substances that are poorly soluble in gastric juice or that slowly dissolve in gastric juice and are used for coating include hydroxypropylcellulose, methacrylic acid copolymer, hydroxypropylmethylcellulose phthalate, ethyl acrylate methyl methyl methacrylate copolymer, and aminoalkylmethacrylate. It is at least one selected from the group consisting of acrylate copolymer, shellac, hydroxyaberite, twain, dextrin, gelatin, arabia gum and sucrose, and is used as a matrix with a substance that is poorly soluble in gastric juice or a substance that gradually dissolves in gastric juice. What is used is hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylcellulose, methylcellulose, carboxymethylcellulose, sodium salt of hydroxypropylmethylcellulose, acrylic acid. Ethyl methyl methacrylate, oil B, such as tridalicelide, fatty acid esters of polyglycerin, etc., and indigestible polysaccharides such as polyvinylpyrrolidone, gelatin, gum arabic, polyethylene glycol, cyclodextrin, and dextrin At least one selected from the group.

The intestinal absorptive anti-inflammatory factor-containing immunological substance of the present invention may be prepared by adding ice cream, candy, baked confectionery, bread, cake, pudding, mousse, jelly, ice confection, sprinkle, gummy, chocolate, caramel, jam, It can be contained in gum or curry.

As described above, the present invention provides a coating or matrix of an anti-inflammatory factor-containing immunological substance with a poorly soluble substance in gastric juice or a substance that gradually dissolves in gastric juice to reduce the amount of the anti-inflammatory factor-containing immunological substance dissolved in the stomach. The aim is to introduce as much of the immunological substance containing an anti-inflammatory factor as possible into the intestine and to enhance its absorption in the intestine. At this time, the form of the anti-inflammatory factor-containing immunological substance may be any of powder, granules, tablets, and tablets. Also, a powdered or granular anti-inflammatory factor-containing immunological substance may be sealed in a capsule coated with an enteric substance on its surface, or an anti-inflammatory factor may be contained in a microcapsule coated with an enteric substance. The immune material may be sealed. Further, those having different thicknesses of the coating layers or those having different component ratios in the matrix may be mixed and used.

Examples of poorly soluble substances in gastric juice or substances that gradually dissolve in gastric juice and used for coating include hydroxypropylcellulose, methacrylate copolymer, hydroxypropylmethylcellulose phthalate, ethyl methacrylate methyl acrylate copolymer, aminoalkyl methacrylate. Examples include acrylate copolymer, shellac, hydroxyapatite, owwain, dextrin, gelatin, gum arabic and sucrose.

In addition, substances that are poorly soluble in gastric juice or that gradually dissolve in gastric juice and are used in the matrix include hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylcellulose, methylcellulose, carboxymethylcellulose, salts of lipoxylmethylcellulose, and acrylic. Examples include oils and fats such as methyl ethyl methacrylate copolymer and tridalicelide, fatty acid esters of polyglycerin, and indigestible polysaccharides such as polyvinylpyrrolidone, gelatin, gum arabic, polyethylene glycol, cyclodextrin and dextrin.

Add the immune substance containing anti-inflammatory factor with enhanced intestinal absorption obtained in this way, and add ice cream, candy, baked confectionery, bread, cake, pudding, muss, jelly, frozen dessert, sprinkle, gummy, Chocolate, caramel, jam, gum or curry can be adjusted.

As described above, the anti-inflammatory factor-containing immunological substance is obtained after defatting and decasein. A substance obtained by mixing a milk protein fraction separated from the obtained whey with a low molecular weight fraction having a molecular weight of 10,000 or less, which does not contain milk fat, lactose, casein, etc. which is usually contained in milk. is there. The effect of heating when milk fat, lactose, casein, etc. is not contained is greater than when milk fat, lactose, casein, etc. is contained. Therefore, when lactose, casein, etc. are not contained, it is needless to say that individual sterilization and sterilization methods are required when lactose, casein, etc. are contained.

The present invention does not necessarily prescribe an anti-inflammatory factor-containing immunological substance as a pharmaceutical, but aims to be used as a so-called health food, adapted to food and drink. As mentioned earlier, anti-inflammatory factor-containing immunological substances are impaired by acids, heat and organic solvents. Because of the presence of stomach acid in the stomach, eating an anti-inflammatory factor-containing immune substance as it is can significantly alter the stomach and impair its effectiveness. In order to maintain its effect in the stomach, the stomach should be as unaltered as possible and dissolved and absorbed only when it reaches the intestine.

A poorly soluble substance in gastric juice or a substance that gradually dissolves in gastric juice and coated or formed with an anti-inflammatory factor-containing immunological substance can be added to food or drink to produce a healthy food. These foods and drinks are useful as health foods that have anti-inflammatory and immune-enhancing effects. Add and prepare an anti-inflammatory factor-containing immunological substance that has improved intestinal absorption, and add ice cream, candy, baked goods, bread, cake, pudding, mousse, jellyfish, frozen dessert, sprinkle, gummy, chocolate, caramel, jam , Gum or wool can be made. BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, embodiments of the present invention will be described. The essence of the present invention is to dissolve the anti-inflammatory factor-containing immunological substance in the stomach by coating or matrixing the anti-inflammatory factor-containing immunological substance with a gastric juice sparingly soluble substance or a substance that gradually dissolves in gastric juice. It seeks to suppress and increase intestinal absorption. In order to coat a poorly soluble substance in gastric juice or a substance that gradually dissolves in gastric juice to an immunological substance containing an anti-inflammatory factor, a powdery, granular or tablet immunological substance containing an anti-inflammatory factor, A method in which a solution in which a substance gradually dissolved in gastric juice is dissolved is directly sprayed, or a solution in which an enteric substance is dissolved in a surface of a capsule or microcapsule in which a powdered or granular immune substance containing an anti-inflammatory factor is sealed is applied. The method of spraying is adopted. Among them, a method of coating a capsule can be suitably used. Coating method (1) The thickness of the coating layer is appropriately selected according to the form of use. For example, when coating shellac,

If it is about 13 g nom ² , it will dissolve in gastric juice in about 2 hours, so this thickness is a good guideline. On the other hand, those having different thicknesses of the coating layer may be mixed and used, or the coating may be not only the whole anti-inflammatory factor-containing immunological substance but also a part thereof.

For the poorly gastric capsule, a method is used in which a substance that is hardly soluble in gastric juice or a substance that gradually dissolves in gastric juice is coated on the surface of the ordinary gelatin capsule film. Examples of substances to be coated on the capsule film surface include hydroxypropyl cellulose, methacrylic acid copolymer, hydroxypropyl methylcellulose phthalate, ethyl acrylate methacrylic acid methyl copolymer, and amino amino J-methacrylate copolymer. , Shellac, hydroxyapatite, twain, dextrin, gelatin, gum arabic and sucrose.

To form a poorly soluble substance in gastric juice or a substance that gradually dissolves in gastric juice into an immune substance containing anti-inflammatory factor and a matrix, slowly dissolve in powdered immunogenic substance containing anti-inflammatory factor and poorly soluble substance in gastric juice or gastric juice Mix well with the substance and fuse the two with a binder such as glucose or viscous cellulose. This is sieved to obtain a powdery matrix. The powdered matrix is granulated and sieved to obtain a granule and a tablet to obtain a tablet. Powder obtained in this way or A granular matrix body may be encapsulated in a capsule.

As a substance which is hardly soluble in gastric juice or a substance which gradually dissolves in gastric juice and is used in the matrix, as described above, hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylcellulose, methylcellulose, carboxymethylcellulose, and lipoxylmethylcellulose can be used. Salts, methyl methacrylate methacrylate copolymer, fats and oils such as tridalicelide, fatty acid esters of polyglycerin, etc., indigestible polysaccharides such as polypinylpyrrolidone, gelatin, arabia gum, polyethylene glycol, cyclodextrin and dextrin. be able to.

As a specific embodiment, a case will be described in which Tween, which is a kind of enteric substance, is coated on a capsule containing a granular anti-inflammatory factor-containing immunological substance. Tween (5 parts by weight) was added to a mixed solution consisting of 80 parts by weight of ethyl alcohol and 15 parts by weight of purified water, and mixed and dissolved at a temperature of 60 "C or lower. The solution was filtered to remove insolubles. On the other hand, a seam capsule made of gelatin in which a granular anti-inflammatory factor-containing immunological substance was put in a seam capsule made of gelatin was prepared. The temperature was sprayed onto the capsule at a rate of 50 gZ at a temperature, and the chain was coated on the capsule containing the granular anti-inflammatory factor-containing immunological substance.

Next, an example in which a cellulose derivative is used as a poorly soluble substance in gastric juice or a substance that gradually dissolves in gastric juice will be described. Hydroxypropyl methylcellulose phthalate is dissolved in a 1: 1 mixture of acetone and ethyl sorbate, and a medium chain fatty acid tridaliceride is used as a plasticizer. The added solution was prepared. This solution is sprayed onto a capsule sealed with an anti-inflammatory factor-containing immunological substance or a bovine inflammatory factor-containing immunological substance using a coating pan or the like by spraying, etc. I got Example

An experiment was conducted to confirm the effect of an anti-inflammatory factor-containing immunological substance coated with a substance that is poorly soluble in gastric juice or a substance that gradually dissolves in gastric juice. Each group consisted of 10 healthy subjects, three groups of subjects, a capsule containing normal whey protein (abbreviated as WP) and whey protein made from milk obtained from immunized cows. That is, a capsule (abbreviated as WPI +) encapsulating an anti-inflammatory factor-containing immunological substance, and a capsule (abbreviated as enteric) treated with twein to make the capsule enteric are included in each group. , And the number of total bacteria and the number of bifidobacteria in the excreted feces of each person were measured. The number of bacteria in each group was averaged, and the ratio of bifidobacteria to the total number of bacteria was calculated. The results are shown in Table 1.

It can be seen that administration of WPI + capsules increased the occupancy of bifidobacteria in the stool. This increase was confirmed two weeks after the start of administration. However, with the administration of WP capsule, the proportion of bifidobacteria began to rise at the third week. Enteric capsules were seen earlier in the trend than WPI + capsules. Table 1 shows the change in the proportion of bifidobacteria after administration relative to the proportion of bifidobacteria before administration. The change in the post-administration ratio when the ratio of the bifidobacteria at the start was set to 1 is expressed as a coefficient. It can be seen that the administration of enteric coated mice increased the proportion of bifidobacteria most.

As described above, the proportion of bifidobacteria in the stool increases with the administration of WPI +. It was also confirmed that the effect was further enhanced by using an enteric capsule.

The sieved WPI + was mixed with the sieved poorly soluble substance in gastric juice or a substance gradually soluble in gastric juice, hydroxypropylmethylcellulose as a binder, talc and a flavoring agent as a lubricant, a sweetener and the like. The mixture was kneaded, granulated, sized, dried, and tableted to obtain tablet-like WPI +. On the surface of this tablet, Coatings can be made as needed.

In the production of granules (fine granules) and microcapsules, known methods can be used. In addition to the method of applying a necessary coat after the production of the microcapsules, a substance that is hardly soluble in gastric juice or a substance that gradually dissolves in gastric juice can be mixed into the capsule shell. Here, a method of coating after wet granulation will be described. Mix and knead the sieved WP I + with the sieved excipients, binders, sweeteners, flavors, etc., granulate, siege, dry and coat. The coating can be coated in one layer or in multiple layers. For example, the first layer is a hydroxypropylmethylcellulose solution, the second layer is an ethylcellulose solution, the third layer is a mixture of hydroxypropylmethylcellulose and triethyl citrate, and the fourth layer is a mixture of hydroxypropylmethylcellulose and titanium oxide. It can be coated with liquid. Industrial applicability

The surface of the capsule or microcapsule enclosing the anti-inflammatory factor-containing immunological substance is made hardly soluble in gastric juice by coating or matrixing the anti-inflammatory factor-containing immunological substance with a substance that is sparingly soluble in gastric juice or a substance that gradually dissolves in gastric juice. Ingestion of a substance or a substance that gradually dissolves in gastric juice produces a significant immune effect. This effect is remarkable compared to the conventionally known ingestion in the form of powders, granules, tablets and capsules of immunological substances containing anti-inflammatory factors.

The number of bacteria is 1 week after the start of X 1 o ⁹ 2 weeks 3 weeks

WP 1 + total bacterial count 43.6 46.4 59.8 60.7

Bifidobacterium 7.03 7.65 12.8 14.4 Bifidobacterium occupancy (%) 16.1 16.5 21.4 23.7 Change (%) 1.0 1.0 1.3 1.5

WP total bacterial count 33.8 33.0 40.0 43.9

Bifidobacterium 4.56 4.56 5.52 9.81 Bifidobacterium occupancy (%) 13.5 13.8 13.8 20.3 Change (%) 1.0 1.0 1.0 1.5 Enteric-soluble total bacteria 39.8 39.7 42.2 54.4

Bifidobacterium 6.09 7.14 9.84 13.8 Bifidobacterium occupancy (%) 15.3 18.0 23.3 25.4 Change (%) 1.0 1.2 1.5 1.7

Claims

The scope of the claims

1. Intestinal absorptive anti-inflammatory factor-containing immunological substance in which an immunological substance containing an anti-inflammatory factor is made into a matrix or coated with a substance that is poorly soluble in gastric juice or a substance that gradually dissolves in gastric juice.

2. The intestinal absorptive anti-inflammatory factor-containing immunological substance according to claim 1, wherein the immunological substance is in the form of a powder, granules, tablets, or pills.

3. An intestinal absorbable anti-inflammatory agent characterized by encapsulating a powdered or granular anti-inflammatory factor-containing immunological substance coated or matrixed with a substance that is poorly soluble in gastric juice or a substance that slowly dissolves in gastric juice. Immune substance containing inflammatory factor.

4. The thickness of the coated poorly soluble gastric fluid or the substance gradually dissolved in gastric juice or the amount of the matrix component of the poorly soluble gastric fluid or the substance gradually dissolved in gastric fluid differs in the amount ratio. 4. The intestinal absorbable anti-inflammatory factor-containing immunological substance according to any one of claims 1 to 3.

5. Use a substance with poorly soluble gastric juice or a substance that slowly dissolves in gastric juice on the surface.

Intestinal absorbable anti-inflammatory factor-containing immunological substance in which a powdered or granular anti-inflammatory factor-containing immunological substance is sealed in a capsule.

6. Intestinal absorptive anti-inflammatory factor-containing immunity in which a powdery or granular anti-inflammatory factor-containing immunological substance is sealed in a microphone-mouth capsule coated on the surface with a substance that is poorly soluble in gastric juice or a substance that gradually dissolves in gastric juice. Substance.

7. Substances that are poorly soluble in gastric juice or that slowly dissolve in gastric juice and are used for coating include hydroxypropylcellulose, methacrylic acid copolymer, hydroxypropylmethylcellulose phthalate, ethyl methacrylate methyl acrylate copolymer, amino acid. Claims 1 to 5 characterized in that there is at least one selected from the group consisting of alkyl methacrylate copolymer, shellac, hydroxyapatite, twain, dextrin, gelatin, arabia gum and sucrose. 7. The intestinal absorptive anti-inflammatory factor-containing immunological substance according to any one of the range 6).

8. Substances that are poorly soluble in gastric juice or that slowly dissolve in gastric juice and are used in the matrix include hydroxypropylcellulose, hydroxypropylmethylcellulose, ethylcellulose, methylcellulose, lipoxymethylcellulose, and lipoprotein. Fats and oils such as salts of xylmethylcellulose, ethyl methacrylate methyl acrylate copolymer, tridalicelide, etc., fatty acid esters of polyglycerin, etc. 7. The intestinal-absorbable anti-inflammatory factor-containing immunological substance according to any one of claims 1 to 6, wherein the immunological substance is at least one selected from the group consisting of saccharides.

9. Ice cream, candy, baked confectionery, bread, cake, prepared by adding the intestinal absorptive anti-inflammatory factor-containing immunological substance according to any one of claims 1 to 8. Puddings, mousses, jellies, frozen desserts, sprinkles, gummi, chiyo three-rate, caramel, jam, gum or curry.