JP2019189543A - Anti-human immunodeficiency virus agent composition - Google Patents

Abstract

To provide an anti-human immunodeficiency virus agent composition that can be supplied readily in large quantity and has an effect of preventing HIV infection.SOLUTION: An anti-human immunodeficiency virus agent composition contains a sulfated polysaccharide containing, for example, one or both of fucoidan and ascophyllan, derived from seaweed such as Ascophyllum nodosum, as an active ingredient, and has an infection control activity on human immunodeficiency virus (HIV).SELECTED DRAWING: Figure 1

Description

  The present invention relates to a novel anti-human immunodeficiency virus agent composition.

  Human immunodeficiency virus (HIV) is a type of retrovirus and the causative agent of acquired immunodeficiency syndrome (AIDS). For the treatment of AIDS, nucleic acid anti-HIV agents such as azidothymidine (AZT) and didanosine (ddI) are used, but they have not been completely cured, and the development of new anti-HIV agents is desired. .

  As an example of a new anti-HIV agent, an anti-HIV agent comprising a composition mainly composed of a polysaccharide derived from a hot water extract of Mycobacterium tuberculosis has been proposed (see Patent Document 1).

International Publication No. 00/53203

  However, since the anti-HIV agent described in Patent Document 1 is mainly composed of a polysaccharide derived from a hot water extract of Mycobacterium tuberculosis, there is a problem that it is difficult to supply a large amount and the production cost is increased. In addition, a method for preventing the onset of AIDS by taking an anti-HIV agent and creating an environment in which HIV does not proliferate in the body is known before the onset of AIDS. The drug to have is not known.

  The present invention has been made in view of such circumstances, and an object of the present invention is to provide an anti-human immunodeficiency virus agent composition that is easy to supply in large quantities and has an effect of preventing HIV infection.

  The present invention that meets the above-described object provides an anti-human immunodeficiency virus agent composition characterized by comprising a seaweed-derived sulfated polysaccharide as an active ingredient and having infection-preventing activity against human immunodeficiency virus (HIV). The present invention solves the above problems.

  In the anti-human immunodeficiency virus agent composition according to the present invention, the seaweed may be Ascophyllum nodosum.

  In the anti-human immunodeficiency virus agent composition according to the present invention, the sulfated polysaccharide may contain one or both of fucoidan and ascofilan.

  ADVANTAGE OF THE INVENTION According to this invention, the anti-human immunodeficiency virus agent composition which has high infection prevention activity with respect to HIV can be obtained from the seaweed which can be obtained in large quantities. The anti-human immunodeficiency virus agent composition of the present invention has the effect of eliminating the need for long-term drug administration for inhibiting the growth of HIV in HIV-infected cells by preventing HIV infection itself. Have. The anti-human immunodeficiency virus agent composition of the present invention has an effect of having high safety because it contains a sulfated polysaccharide derived from seaweed with dietary experience as an active ingredient.

  The present invention also provides a novel method for preventing HIV infection.

It is a figure which shows the experimental result of the infection prevention activity of HIV-1 in a human CD4 positive T cell.

  Subsequently, an embodiment of the present invention will be described to provide an understanding of the present invention. The anti-human immunodeficiency virus agent composition (hereinafter sometimes abbreviated as “composition”) according to an embodiment of the present invention contains a seaweed-derived sulfated polysaccharide as an active ingredient.

<Seaweed>
As the seaweed used as a raw material for the active ingredient of the composition, any one containing a sulfated polysaccharide having an infection-preventing activity against HIV can be used without particular limitation. Specific examples of seaweed include kombu, wakame, mekabu, mozuku, akamoku, honda walla, ascofilum nodsum and the like.

<Sulfated polysaccharide>
The sulfated polysaccharide serving as an active ingredient of the composition is a compound that is contained in seaweed and in which at least a part of the alcoholic hydroxyl group of the polysaccharide is sulfated. Specific examples thereof include sulfated β-1,3- Examples include xylan, sulfated β-1,4-xylan, porphyran, fucoidan, and ascofilan. The sulfated polysaccharide which is an active ingredient of the composition may be one type of sulfated polysaccharide or a mixture containing any two or more types in any proportion.

  The sulfated polysaccharide is isolated from seaweed as a raw material using any known method such as hot water extraction. Examples of preferred sulfated polysaccharides include fucoidan and ascofilan.

<Ascofilum Nodsum>
Ascofilum nodosum is a kind of large brown algae that breeds in the coastal regions of Scandinavia, and is used as a livestock feed and plant growth promoter. Ascophyllum nodsum contains water-soluble polysaccharides such as alginic acid, fucoidan, mannit, laminarin, and ascofilan in addition to amino acids, vitamins, ash, polyphenols, and various minerals. Ascofilan is a polysaccharide unique to Ascophyllum nodsum and is similar to fucoidan in that it contains L-fucose and sulfate groups, but the composition of sugar residues is different, and it has the ability to induce apoptosis and certain types of cancer cells. It is known to have antitumor activity against.

<Ascophyllum / Nodsum extract>
Any known method can be used as a means for separating the compound contained in Ascophyllum nodsum, and a preferable method includes solvent extraction, and particularly preferable is solvent extraction using water as a solvent. As an as raw material for extraction, Ascophilum nodsum can be from any source. In addition, since raw algal bodies have a high water content, it is preferable to use a dried algal body from which salt has been removed by washing with water as a raw material for extraction. The average composition of the dried powder of Ascophyram nodsum used as the raw material for extraction is 5 to 10% protein, 45 to 60% carbohydrate, 17 to 20% ash, 2 to 4% lipid, and 10 to 12% moisture. In addition, as a pretreatment, the ascophyllum / nodsum dry powder may be treated with an acid aqueous solution to remove acid-soluble components.

  Extraction from Ascophilum nodosum is performed using, for example, 10 to 100 times (mass ratio) of water as dry powder of Ascophilum nodosum. Ascofilan is extracted into water together with alginic acid and the like when the water to which the dry powder of Ascophylam nodosa is added is stirred at room temperature for 12 to 24 hours. If necessary, water may be newly added, and extraction may be further performed at 100 ° C. for 12 to 24 hours. The crude extract thus obtained contains other polysaccharides, proteins, carbohydrates, lipids, ash, minerals, pigments, polyphenols and the like. Since these do not cause any harm to the human body and animals, when the Ascophyram nodosa extract is added to food or feed, it may be used as it is without being separated.

  In order to obtain fucoidan and ascofilan having high purity that can be used as a raw material of the composition, purification can be performed by the method described below. Alginic acid contained in the extract can be removed by adding hydrochloric acid or the like to lower the pH of the extract and filtering the produced precipitate. Alternatively, alginic acid lyase may be added to selectively hydrolyze only alginic acid, and the hydrolyzate of alginic acid may be removed by dialysis. When 50% ethanol is added to the extract, the precipitate obtained is dissolved in water, desalted by dialysis, and lyophilized to obtain almost pure ascofilan. If further purification is required, reprecipitation may be performed from water-ethanol.

  The 50% ethanol-soluble fraction described above contains fucoidan, and high-purity fucoidan can be obtained by fractional purification as necessary.

<Fucoidan>
Fucoidan is a sulfated polysaccharide in which a large number of L-fucose is mainly bonded and sulfuric acid is bonded to at least a part of the L-fucose residue to form a sulfate ester. In addition to the L-fucose residue, a glucuronic acid residue or a galactose residue may be bound. Fucoidan may be isolated from Ascofilum nodsum together with Ascofilan described later, but may be isolated from other seaweeds such as kombu, wakame, mozuku, and akamoku.

<Ascofilan>
Ascophyrane, which contains a sulfate group and a D-glucuronic acid group and is a polyanion, may form a salt with any cation that is water-soluble and does not inhibit the HIV infection-suppressing activity. Examples of such cations include sodium ions, potassium ions, magnesium ions, calcium ions, ammonium ions, and the like.

  Ascofilan that can be used as an active ingredient of the composition is any sulfated polysaccharide extracted from Ascophilum nodsum and having D-glucuronic acid as a main component in addition to L-fucose and D-xylose. There are no particular restrictions on the molecular weight. The sulfate group content and the uronic acid content are not particularly limited, but those having a sulfate group content of 7 to 15% by weight measured by the turbidimetric method using barium sulfate are preferred, and uron measured by the carbazole sulfate method is used. Those having an acid content of 20 to 25% by weight are preferred.

<Anti-human immunodeficiency virus agent composition>
Ascophyllum nodosum extract or fucoidan and / or ascofilan isolated therefrom is used as a pharmaceutical composition which is an example of an anti-human immunodeficiency virus agent composition by mixing with a pharmaceutically acceptable carrier or the like. be able to. Examples of the dosage form of the pharmaceutical composition for humans or animals include oral administration and parenteral administration (for example, intravenous administration, intramuscular administration, subcutaneous administration, inhalation administration) and the like. It is difficult to determine unambiguously and appropriately set depending on the form, administration method, purpose of use and age, weight, and symptoms of the administration subject applied to it, but in the case of oral administration to humans, The amount of the active ingredient contained (Ascophyllum nodsum extract) is preferably 10 to 1000 mg / kg per day, more preferably 20 to 200 mg / kg per day. In the case of oral administration to humans, the administration frequency and interval are once a day (24-hour intervals). Of course, since the dose and the administration interval vary depending on various conditions, an amount smaller than the above-mentioned dose may be sufficient or may be necessary beyond the range.

  When prepared as an orally administered preparation, it can be prepared in the form of tablets, granules, powders, capsules, coatings, liquids, suspensions, etc. It can be prepared in the form of drops, suppositories and the like. Any known method can be used for formulation. For example, Ascofilum nodosum extract and a pharmaceutically acceptable carrier or diluent, stabilizer, and other desired additives can be blended into the desired dosage form described above.

  The composition may be a food product. As food containing the composition, the ascophyllum / nodsum extract prepared as it is, added to other foods, or any form normally used for food or health food such as capsules, tablets, etc. Can do. When ingested or administered in foods, mix with excipients, extenders, binders, thickeners, emulsifiers, colorants, flavors, food additives, seasonings, etc. Accordingly, it can be formed into a powder, granule, tablet or the like. Moreover, it can mix suitably in a food raw material, can prepare a foodstuff, and can use it as a functional food which has the infection prevention activity of HIV.

Next, examples carried out for confirming the effects of the present invention will be described.
Example 1: Purification of Ascofilan and Fucoidan derived from Ascophilum nodosum After washing Ascophilum nodosum with dilute citric acid water, hot water extraction (90 ° C.) was performed. Next, neutralization was performed after removing insolubles by centrifugation and filtration. Thereafter, acid was added to precipitate alginic acid to roughly remove alginic acid, and then the remaining alginic acid was decomposed and removed by alginate lyase treatment. Ethanol was added to the resulting solution, and ascofilan (“As” in FIG. 1) was obtained from the precipitate, and fucoidan (“A-Fu” in FIG. 1) was obtained from the supernatant.

Example 2: Examination of the effect of sulfated polysaccharides on the initial infection of HIV-1 to human CD4-positive T cells MT-4 cells (human CD4-positive T cells) were added to the wells at a density of 1 × 10 ⁶ cells. , Sulfated polysaccharide (Ascofilan: As, alginate oligomer: Ao, Fucoidan manufactured by Sigma: S-Fu, Fucoidan derived from Ascophilum Nodsum: A-Fu, porphyran: Po, dextran sulfate: De-S, dextran: De, concentration : 100 μg / mL) for 1 hour. Thereafter, the HIV-1 R9 strain was added and incubated for 48 hours. The degree of infection with HIV was determined by quantifying HIV p24 protein in cell lysates (Western blotting), infecting HIV released into MT-4 cell culture supernatant into TZM-bl cells, and by luciferase method. evaluated.

  The results are shown in FIG. In FIG. 1, “Neg” indicates a negative control incubated without addition of HIV, and “Con” indicates a positive control incubated without addition of polysaccharide. From the results of FIG. 1, it was confirmed that ascofilan and fucoidan suppress early infection of HIV to MT-4 cells.

Claims (3)

  An anti-human immunodeficiency virus agent composition comprising a sulfated polysaccharide derived from seaweed as an active ingredient and having an infection-preventing activity against human immunodeficiency virus (HIV).   The anti-human immunodeficiency virus agent composition according to claim 1, wherein the seaweed is Ascophyllum nodosum.   The anti-human immunodeficiency virus agent composition according to claim 1 or 2, wherein the sulfated polysaccharide contains one or both of fucoidan and ascofilan.