JP3920954B2 - Manufacturing method of anti-ulcer agent - Google Patents

Description

【００３０】
試験例２
実施例１で得られたフコイダンについて、試験例１と同様の試験を行なった。その結果を表２に示す。
【００３１】
【表２】

【００３２】
試験例３
実施例２で得られたフコイダンについて、試験例１と同様の試験を行なった。その結果を表３に示す。
【００３３】
【表３】

【００３４】
試験例４
実施例３で得られたフコイダンについて、試験例１と同様の試験を行なった。その結果を表４に示す。
【００３５】
【表４】

【００３６】
試験例５
実施例４で得られたフコイダンについて、試験例１と同様の試験を行なった。その結果を表５に示す。
【００３７】
【表５】

【００３８】
【発明の効果】
上述のように、本発明によればいかなるモズクからも抗潰瘍活性の優れたフコイダンが得られるようになり、フコイダンからなる抗潰瘍剤を安価に、且つ容易に製造することが可能になる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a method for producing an anti-ulcer agent containing fucoidan derived from mozuku as an active ingredient.
[0002]
[Prior art]
Fucoidan is a polysaccharide mainly composed of fucose and having a molecular weight of around 100,000, and a part of fucose unit is sulfated. Fucoidan has been confirmed to exhibit an anti-ulcer action and can be used as a component of an anti-ulcer agent.
[0003]
Fucoidan is abundant in Mozuku (Phaeophyceae Chordariales nemacystus), a seaweed belonging to the brown algae, and is easy to extract. This is advantageous (Japanese Patent Laid-Open No. 7-138166).
[0004]
As a method for extracting fucoidan from mozuku, there is a hot water extraction method. In particular, extraction with hot water to which an acid is added is excellent in terms of extraction efficiency, yield and the like. An outline of a method for extracting fucoidan by an acid-added hot water extraction method, purifying it, and using it as an anti-ulcer agent is as follows.
[0005]
Extraction method: Mozuku alga bodies are suspended in about 1 to 3 times the wet weight of water and diluted hydrochloric acid is added to adjust the pH to 2-4, preferably 2-3. Next, the mixture is heated to about 50 ° C. or higher, preferably about 80 to 100 ° C. to elute fucoidan. The precipitate is then centrifuged to remove the precipitate and the supernatant is neutralized with caustic soda solution.
[0006]
Purification method: Perform molecular weight fractionation by ultrafiltration, dialysis, etc. to remove salts and low molecular weight impurities. It can be purified by gel filtration, ion exchange chromatography or the like. Finally, it is concentrated and freeze-dried to obtain powdered fucoidan.
Fucoidan administered orally as an anti-ulcer agent is considered to enter the digestive tract and reach the affected area to effect ulcer healing.
[0007]
[Problems to be solved by the invention]
The anti-ulcer activity of fucoidan extracted from mozuku varies greatly depending on the lot of mozuku as a raw material, and it has been found that in rare cases it may not show any activity. This variation in activity seems to be complicatedly related to the maturity of the raw mozuku, the harvest year, etc. Therefore, it was difficult to produce an anti-ulcer agent of a certain quality using mozuku as a raw material.
[0008]
Accordingly, an object of the present invention is to make it possible to obtain fucoidan having a small fluctuation range of anti-ulcer activity from mozuku extract, thereby facilitating the production of an anti-ulcer agent comprising fucoidan.
[0009]
[Means for Solving the Problems]
The present invention that has succeeded in achieving the above object is to produce an anti-ulcer agent using fucoidan extracted from mozuku. The present invention provides a method for producing an anti-ulcer agent, which is characterized by being converted to a salt form .
[0010]
The ion exchange treatment in the present invention can be preferably carried out by electrodialysis, acid washing using an ultrafiltration membrane, ion exchange resin treatment, or any of these treatments followed by neutralization treatment. The processing means is not limited to these.
[0011]
When fucoidan extracted from mozuku is subjected to ion exchange treatment to convert its sulfate ester group to the free acid form or alkali metal salt form, fucoidan that does not exhibit anti-ulcer activity or weak activity without this treatment also has strong activity. As shown. Therefore, fucoidan exhibiting strong anti-ulcer activity has its sulfate group mostly in the form of free acid or alkali metal salt, and fucoidan as extracted from mozuku has some or most of its sulfate group. It is conceivable that a polyvalent metal may be bound to the portion, which inhibits the expression of antiulcer activity, but details have not been confirmed yet.
[0012]
DETAILED DESCRIPTION OF THE INVENTION
When producing an anti-ulcer agent according to the present invention, the method for extracting fucoidan from mozuku is not particularly limited, but the acid-added hot water extraction method described above is most advantageous.
When an extract containing fucoidan is obtained, the ion exchange treatment according to the present invention is performed at an arbitrary stage of the purification step for removing low molecular weight water-soluble components, salts and the like therefrom. The ion exchange treatment can be performed by electrodialysis, acid washing using an ultrafiltration membrane, ion exchange resin treatment, or any of these treatments followed by neutralization treatment.
[0014]
In electrodialysis, a solution to be treated is allowed to flow between a cation exchange membrane that allows only cations to pass through and an anion exchange membrane that allows only anions to pass through. Those having a relatively small radius are removed from the solution to be treated by permeating through the ion exchange membrane. Although it is a well-known means for desalting or concentrating salts, a solution of a polymer electrolyte such as fucoidan is used because the ion exchange membrane is dense and organic substances other than those having a very low molecular weight are not permeated. When this treatment is performed, metal ions bonded to fucoidan are removed in parallel with the removal of the inorganic salts that coexist as impurities. Fucoidan that does not permeate the ion exchange membrane remains in the solution to be treated in the form of free acid. The progress of the process can be monitored by continuously measuring the electrical conductivity.
[0015]
Acid washing using an ultrafiltration membrane can be performed in combination with purification by desalting and molecular weight fractionation by ultrafiltration. That is, if the fucoidan-containing extract is ultrafiltered using an ultrafiltration membrane whose molecular weight cut off is sufficiently smaller than the molecular weight of fucoidan, only low molecular weight impurities such as inorganic salts can be permeated and removed. At this time, if diluted hydrochloric acid is added to the non-permeated liquid in which fucoidan is concentrated and ultrafiltration is performed again, metal ions released from fucoidan permeate the membrane, and fucoidan has its sulfate ester group remaining in the form of free acid.
[0016]
The ion exchange by the ion exchange resin treatment can be performed by a conventional method using an H-form strongly acidic ion exchange resin, and the sulfate group of fucoidan can be most surely changed to the free acid form.
[0017]
Whichever ion exchange means is employed, an alkali metal salt can be obtained by neutralizing fucoidan in a free acid form with an alkali metal hydroxide such as caustic soda or caustic potash.
[0018]
There is no significant difference in the anti-ulcer activity of fucoidan depending on whether the sulfate group is in the free acid form or the alkali metal salt form, but the salt form is superior in terms of storage stability.
[0019]
Fucoidan that has been sufficiently purified and the sulfate ester group is in the form of a free acid or alkali metal salt can be combined with any auxiliary agent or other agent to form an anti-ulcer agent by any formulation means.
[0020]
【Example】
Hereinafter, the present invention will be described with reference to examples and comparative examples. In each case, it was previously confirmed that the mozuku used as the extraction material for fucoidan could only produce fucoidan that showed almost no anti-ulcer activity by the method of Comparative Example 1.
[0021]
Comparative Example 1
1 liter of tap water was added to 1 kg of mozuku salted algae, pH was adjusted to 3 with 1N-HCl, and the mixture was heated to 95 ° C. for 1 hour to extract fucoidan. After cooling, adjust the pH to 6 with caustic soda, centrifuge at 10,000G for 10 minutes to remove the extraction residue, and use the obtained extract with an ultrafiltration membrane with a molecular weight cut off of 6,000 And filtered. The non-permeate in which fucoidan was concentrated was ultrafiltered again by adding 2 liters of deionized water, and finally concentrated under reduced pressure and lyophilized. 21 g of powdered fucoidan (purity 75% by weight) was obtained.
[0022]
Example 1
The same mozuku used in Comparative Example 1 was extracted in the same manner as in Comparative Example 1, and centrifuged to obtain an extract. Concentrate the resulting extract to about 500 ml under reduced pressure and treat it with an electrodialyzer / microacylator G3 type (all manufactured by Asahi Kasei Kogyo Co., Ltd.) equipped with an ion exchange membrane / Aciplex cartridge AC-110-400. Thus, the metal ions bound to the low molecular electrolyte and fucoidan were removed. After the treatment, the solution was concentrated under reduced pressure and freeze-dried to obtain 25 g of powdered fucoidan (purity 60% by weight).
[0023]
Example 2
300 kg of the same mozuku used in Comparative Example 1 was extracted in the same manner as in Comparative Example 1, and centrifuged to obtain an extract. The obtained extract was filtered using an ultrafiltration membrane having a fractional molecular weight of 6,000 to remove low molecular weight impurities. Next, 200 liters of deionized water and hydrochloric acid were added to the non-permeate enriched with fucoidan to obtain a diluted solution of pH3, followed by ultrafiltration using the same ultrafiltration membrane. By further repeating the acid washing treatment of ultrafiltration using a diluted solution of pH 3 twice, adding 200 liters of deionized water and ultrafiltration twice, and finally concentrating and freeze-drying. Thus, 4.5 kg (purity 76% by weight) of powdered fucoidan was obtained.
[0024]
Example 3
In the same manner as in Comparative Example 1, 200 g of powdered fucoidan was produced, dissolved in 10 liters of deionized water, and passed through a column of H-form strongly acidic cation exchange resin / Diaion SK1B. The column-passed solution was neutralized with an aqueous caustic soda solution, concentrated under reduced pressure, and lyophilized to obtain 200 g of fucoidan having a Na ester salt form (purity: 70% by weight).
[0025]
Example 4
150 g of powdered fucoidan produced in Example 2 was dissolved in deionized water, adjusted to pH 6 with caustic soda, concentrated and freeze-dried, and 152 g of fucoidan having a Na salt form as a sulfate group (purity 76). % By weight).
[0026]
Test example 1
The fucoidan obtained in the comparative example was tested for anti-ulcer activity using acetic acid-induced ulcer. The test method is as follows (the same applies to the following examples).
[0027]
Test method: For acetic acid-induced ulcers, 8-week-old SD rats (body weight 250-300 g; 10 per group) were laparotomized under Nembutal anesthesia, the stomach was taken out and 20% acetic acid was added to the gastric body submucosa. Generate by injecting 03 ml. From the next day to the 10th day after the operation, the fucoidan is orally administered, and on the 11th day, the stomach is removed to measure the area of ulcer formation (major axis x minor axis). Yes (displays mean ± standard deviation). Further, the healing rate is calculated by the following formula. During the test period, food and water are ad libitum.
Curing rate (%) = (1-ulcer index of fucoidan administration group / ulcer index of control group) × 100
[0028]
The test results are shown in Table 1. [0029]
[Table 1]

[0030]
Test example 2
The fucoidan obtained in Example 1 was tested in the same manner as in Test Example 1. The results are shown in Table 2.
[0031]
[Table 2]

[0032]
Test example 3
The fucoidan obtained in Example 2 was tested in the same manner as in Test Example 1. The results are shown in Table 3.
[0033]
[Table 3]

[0034]
Test example 4
The fucoidan obtained in Example 3 was tested in the same manner as in Test Example 1. The results are shown in Table 4.
[0035]
[Table 4]

[0036]
Test Example 5
The fucoidan obtained in Example 4 was tested in the same manner as in Test Example 1. The results are shown in Table 5.
[0037]
[Table 5]

[0038]
【The invention's effect】
As described above, according to the present invention, fucoidan having excellent anti-ulcer activity can be obtained from any mozuku, and an anti-ulcer agent comprising fucoidan can be easily produced at low cost.

Claims (2)

In producing an anti-ulcer agent using fucoidan extracted from mozuku, an anti-ulcer characterized by converting the fucoidan sulfate group into a free acid form or an alkali metal salt form by subjecting the fucoidan to an ion exchange treatment. Manufacturing method. The production method according to claim 1, wherein the ion exchange treatment is performed by electrodialysis, acid washing using an ultrafiltration membrane, ion exchange resin treatment, or any one of these treatments followed by neutralization treatment .