WO2024185742A1

WO2024185742A1 - Prophylactic or therapeutic agent for disease induced by parasite

Info

Publication number: WO2024185742A1
Application number: PCT/JP2024/008052
Authority: WO
Inventors: 啓輔菅沼; 潔北; 雅一山本
Original assignee: 国立大学法人北海道国立大学機構; 国立大学法人長崎大学; 株式会社ミトコンドリア研究所
Priority date: 2023-03-03
Filing date: 2024-03-04
Publication date: 2024-09-12
Also published as: JP2024124790A

Abstract

Provided are a prophylactic or therapeutic agent for a disease induced by a parasite, the agent being capable of reducing the dosage of ascofuranone or glycerol, a pharmaceutical, a food, a beverage, a feed and a pharmaceutical composition.　This prophylactic or therapeutic agent for a disease induced by a parasite comprises, as an active ingredient, ascofuranone-producing fungal cells that contain ascofuranone produced by per se.

Description

Preventive or therapeutic agent for diseases caused by parasites

The present invention relates to a preventive or therapeutic agent for diseases caused by parasites.

Diseases caused by parasites include African sleeping sickness (human African trypanosomiasis), Nagana disease (animal African trypanosomiasis), Chagas disease (American trypanosomiasis), and leishmaniasis, which are caused by protozoa, as well as echinococcosis, which is caused by helminths.

Ascofuranone is effective as a preventive and therapeutic agent for diseases caused by African trypanosoma protozoa classified in the Trypanosomatidae family and the Trypanosoma genus, such as Trypanosoma brucei and Trypanosoma vivax, and it has been reported that its effectiveness against Trypanosoma brucei is enhanced when used in combination with glycerol (see, for example, Patent Documents 1 and 2, and Non-Patent Documents 1 to 3). When trypanosoma protozoa such as Trypanosoma brucei and Trypanosoma vivax invade the mammalian body, they synthesize ATP mainly in the glycolytic pathway in glycosomes, which requires the regeneration of NADH catalyzed by trypanosomal alternative oxidase (TAO). Ascofuranone inhibits the activity of TAO at low concentrations, and therefore has a strong antitrypanosomal effect. Ascofuranone has also been shown to be potentially effective against Chagas disease, leishmaniasis, and echinococcosis (see Patent Document 3 and Non-Patent Document 4).

In order to put ascofuranone into practical use, it is necessary to be able to steadily supply it on an industrial scale, but because total chemical synthesis is difficult, research and development has been conducted on methods for mass production of ascofuranone using microorganisms (see, for example, Patent Documents 4 and 5). A method for purifying ascofuranone is described in Patent Document 6, but only small amounts of ascofuranone could be purified from a 70L culture solution of a microorganism of the genus Ascochyta (later identified as Acremonium egyptiacum), with purification using a silica gel column yielding 5 g and purification using the recrystallization method yielding 3 g, making it necessary to improve the yield in order to ensure a stable supply of ascofuranone.

Japanese Patent Application Publication No. 9-165332 Patent No. 4468638 Patent No. 4606841 International Publication No. 2018/207928 JP 2019-103400 A Special Publication No. 56-25310

Ascofuranone can be administered alone to treat African trypanosomiasis model mice, but large amounts of ascofuranone must be administered to have a therapeutic effect. When treating large animals such as cows and horses, it is difficult to administer large amounts, so the amount of ascofuranone administered must be reduced.

In addition, by combining it with glycerol, the effectiveness of ascofuranone is enhanced, making it possible to reduce the dosage of ascofuranone, but to increase the effectiveness of ascofuranone, large amounts of glycerol must be administered. To alleviate the symptoms of glycerol poisoning, the dosage of glycerol must be reduced.

Therefore, there is a need for a preventive or therapeutic agent for diseases caused by parasites that can reduce the dosage of ascofuranone or glycerol.

The object of the present invention is to provide a preventive or therapeutic agent, medicine, food, drink, feed, and pharmaceutical composition for diseases caused by parasites that can reduce the dosage of ascofuranone or glycerol.

To achieve the above object, the present invention provides the following preventive or therapeutic agents, medicines, foods and beverages, feeds, and pharmaceutical compositions for diseases caused by protozoa.

In the present invention, the expression X (number) to Y (number) means greater than or equal to X and less than or equal to Y, unless otherwise specified.

[1] A preventive or therapeutic agent for a disease caused by a parasite, comprising as an active ingredient an ascofuranone-producing fungus that contains ascofuranone produced by the fungus.
[2] The preventive or therapeutic agent according to [1] above, wherein the parasite is Trypanosoma congolense.
[3] The preventive or therapeutic agent according to [1] above, wherein the parasite is Trypanosoma evansi.
[4] The preventive or therapeutic agent according to [1] above, wherein the disease is trypanosomiasis.
[5] The preventive or therapeutic agent described in [1], wherein the ascofuranone-producing microorganism is a microorganism of the genus Acremonium.
[6] The preventive or therapeutic agent described in [1], wherein the ascofuranone-producing bacterial cell is a bacterial cell that does not produce ascochlorin.
[7] The preventive or therapeutic agent according to [1], which contains glycerol.
[8] The preventive or therapeutic agent according to the above [1], which is for oral administration.
[9] A pharmaceutical comprising the preventive or therapeutic agent according to any one of [1] to [8] above.
[10] A food or drink comprising the preventive or therapeutic agent according to any one of [1] to [8] above.
[11] A feed comprising the preventive or therapeutic agent according to any one of [1] to [8] above.
[12] A pharmaceutical composition comprising, as an active ingredient, an ascofuranone-producing bacterial cell that contains ascofuranone produced by the bacterial cell itself.

The present invention provides a preventive or therapeutic agent, medicine, food or drink, feed, and pharmaceutical composition for diseases caused by parasites that can reduce the dosage of ascofuranone or glycerol.

[Preventive or therapeutic agent for diseases caused by parasites]
A preventive or therapeutic agent for a disease caused by a parasite according to an embodiment of the present invention (hereinafter referred to as "this embodiment") contains ascofuranone-producing bacteria containing ascofuranone produced by the bacteria as an active ingredient.

(Ascofuranone-producing bacteria)
The ascofuranone-producing bacterial cell used in this embodiment contains ascofuranone that it itself produces.

The ascofuranone-producing microorganism may be any microorganism (strain) capable of producing ascofuranone, and is not particularly limited, but may be, for example, a microorganism belonging to the genus Acremonium. Examples of microorganisms belonging to the genus Acremonium that produce ascofuranone include Acremonium egyptiacum (Acremonium sclerotigenum has been reclassified as Acremonium egyptiacum) and Acremonium sp. isolated from marine sponges (see J. Nat. Prod. 2009, 72, 270-27). Alternatively, a highly safe filamentous fungus such as Aspergillus sojae or Aspergillus oryzae can be used by introducing the ascofuranone biosynthesis gene into the fungus to heterologously produce ascofuranone (see J Gen Appl Microbiol. 2022 Jun 20;68(1):10-16). In addition, it is preferable that the ascofuranone-producing fungus is a fungus that does not produce compounds such as ascochlorin that are toxic to mammals.

Furthermore, the ascofuranone-producing bacteria is not particularly limited, but is preferably a bacteria containing 5% by mass or more of ascofuranone in the bacteria (dried matter). The ascofuranone content in the dried bacteria is preferably 10% by mass or more, more preferably 15% by mass or more, 20% by mass or more, 25% by mass or more, 30% by mass or more, 35% by mass or more, 40% by mass or more, 45% by mass or more, 50% by mass or more, 55% by mass or more, 60% by mass or more, 65% by mass or more, or 70% by mass or more. The amount of ascofuranone contained in the dried bacteria can be measured, for example, by immersing a certain weight of the dried bacteria in a certain amount of an organic solvent such as acetone or methanol to extract ascofuranone, and then calculating the concentration of ascofuranone contained in the organic solvent by analyzing it by LC or the like.

Ascofuranone-producing bacteria can be obtained by culturing bacteria capable of producing ascofuranone under culture conditions that produce ascofuranone, followed by sterilization (for example, heating at about 65 to 120°C for about 10 to 30 minutes), and recovering the bacteria by centrifugation at 500 to 10,000 g or the like, or filtration through filter paper or filter cloth. The yield of ascofuranone contained in the bacteria after recovery of the bacteria is preferably 60% or more, and more preferably 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, or 95% or more, assuming that the ascofuranone after the end of cultivation is 100%. In addition, after recovery of the bacteria, it is preferable to carry out a dry heat treatment or freeze-drying treatment to obtain a dried product. The yield of ascofuranone contained in the dried cells is preferably 60% or more, more preferably 65% or more, 70% or more, 75% or more, 80% or more, 85% or more, 90% or more, or 95% or more, when the ascofuranone after the end of the culture is taken as 100%. In addition, the dried cells are preferably pulverized into a powder of a particle size suitable for subsequent administration using a mortar, a grinder, or the like. For example, it can be obtained by the method described in Patent Document 4 (International Publication No. 2018/207928) or Patent Document 5 (JP Patent Publication No. 2019-103400). Specifically, for example, an ascofuranone-producing fungus is prepared by using an ascG-disrupted strain of Acremonium sclerotigenum (now called Acremonium egyptiacum after reclassification) F-1392 strain in which the ascI gene is highly expressed (ΔascG-I strain), culturing this in a medium (liquid medium or solid medium), sterilizing it, and recovering it, thereby obtaining an ascofuranone-producing fungus that contains a high content of ascofuranone and does not contain ascochlorin. The production amount of ascofuranone (production amount per 1 L of culture medium) is preferably 0.5 g/L or more, and more preferably 1 g/L or more.

In this embodiment, in addition to using the ascofuranone-producing bacterial cells recovered after sterilization as they are, the recovered ascofuranone-producing bacterial cells can also be used as a "mixture containing bacterial components and products such as ascofuranone" obtained by treating the recovered ascofuranone-producing bacterial cells with enzymes or physical means. It also includes a case where the bacterial cells are not recovered and the cells are used as a "culture" containing the ascofuranone-producing bacterial cells as well as a medium. These may be used alone or in combination.

(Diseases caused by parasites)
The parasite-induced disease according to the present embodiment is, for example, a disease caused by a parasite belonging to the genus Trypanosoma, the genus Leishmania, or the genus Echinococcus. Ascofuranone has anti-trypanosomiasis activity by inhibiting the alternative oxidase (TAO) of Trypanosoma protozoa at a concentration of less than nM order or at a low concentration of nM order.

The protozoa belonging to the Trypanosoma genus include Trypanosoma congolense, Trypanosoma evansi, Trypanosoma brucei brucei, and Trypanosoma equipardum, which cause trypanosomiasis. soma equiperdum, Gambian trypanosoma (Trypanosoma brucei gambiense), Rhodesian trypanosoma (Trypanosoma brucei rhodesience), Trypanosoma vivax, and Trypanosoma cruzi.

Examples of trypanosomiasis include African sleeping sickness (African trypanosomiasis, hypnotic disease, sleeping sickness, human African trypanosomiasis), Nagana disease (animal African trypanosomiasis), dourine, Surra disease, and Chagas disease (American trypanosomiasis). The main protozoa that cause African sleeping sickness are Trypanosoma brucei gambiense and Trypanosoma brucei rhodesience, while the main protozoa that cause Nagana disease are Trypanosoma congolense, Trypanosoma vivax, Trypanosoma brucei brucei, and so on, and these are sometimes called African trypanosoma protozoa. The main protozoan that causes dourine is Trypanosoma equiperdum, and the main protozoan that causes surra disease is Trypanosoma evansi. The main protozoan that causes Chagas disease is Trypanosoma cruzi, sometimes called the American trypanosome.

Protozoa belonging to the Leishmania genus include Leishmania major. Helminths belonging to the Echinococcus genus include Echinococcus granulosus, E. multilocularis, E. vogeli, and E. oligarthrus.

(Preventive or therapeutic agent)
The preventive or therapeutic agent of this embodiment may contain only ascofuranone-producing bacteria as the active ingredient, or may also contain pharmaceutical additives or carriers, additives or additives for food and beverages, feed additives, etc.

The preventive or therapeutic agent according to this embodiment preferably contains glycerol as an additive in order to enhance the preventive or therapeutic effect of ascofuranone against diseases caused by parasites. The amount of glycerol added can be adjusted as needed, and can be added, for example, as a PBS (phosphate buffer solution) solution containing glycerol at a concentration of 20 to 80 w/w%, preferably 30 to 75 w/w%, and more preferably 35 to 70 w/w%.

As pharmaceutical additives or carriers, any additives or carriers used in pharmaceutical manufacturing can be used. Examples include excipients, diluents, wetting agents, suspending agents, emulsifiers, dispersing agents, adjuvants, sweeteners, colorants, flavoring agents, buffers, preservatives, binders, stabilizers, pH adjusters, disintegrants, sustained-release agents, enteric coating bases, lubricants, adsorbents, gloss agents, etc., and one or more of these can be selected as appropriate depending on the desired pharmaceutical dosage form, etc.

As additives for food and beverages, any additives used in the manufacture of food and beverages can be used. Examples include emulsifiers, antioxidants, thickeners, moisture-proofing agents, preservatives, excipients, vitamins, minerals, sweeteners, acidulants, seasonings, colorants, and flavorings. One or more of these can be selected as appropriate depending on the desired food and beverage.

Feed additives that can be used are those that are added to, mixed with, infiltrated into, or used in other ways with the aim of "preventing deterioration in feed quality," "supplementing the nutritional components and other active ingredients in feed," and "promoting the effective use of the nutritional components contained in feed." Examples include antioxidants, fungicides, binders, emulsifiers, regulators, amino acids, etc., vitamins, minerals, color enhancers (pigments), synthetic antibacterial agents, antibiotics, flavorings, taste enhancers, enzymes, probiotics, and organic acids. One or more of these can be selected as appropriate depending on the desired feed.

The preventive or therapeutic agent according to this embodiment is not particularly limited in the route of administration and may be administered orally or parenterally, but is preferably administered orally. For example, it may be administered orally in the form of tablets, capsules, granules, powders, pellets, liquids, syrups, or jellies. It may also be administered rectally as a suppository, or subcutaneously, intramuscularly, or intravenously as an injection.

The subjects to which the preventive or therapeutic agent according to this embodiment is administered are humans and non-human animals. Non-human animals are preferably mammals and birds, including livestock, poultry, and pets.

The dosage and frequency of administration of the preventive or therapeutic agent according to this embodiment can be adjusted as appropriate depending on the subject, body weight, age, purpose, severity of symptoms, route of administration, dosage form, whether or not glycerol is added, whether or not a known preventive or therapeutic agent for a disease caused by a parasite is used in combination, etc. For example, the administration concentration, administration method, and frequency of administration can be appropriately determined so that the concentration of ascofuranone in the blood reaches a concentration that is effective against the target parasite. For example, Japanese Patent Publication No. 5960300 shows that the minimum inhibitory concentration (MIC) of ascofuranone in Trypanosoma brucei rhodesiensis is 0.0019 nM in the presence of glycerol and 0.033 nM in the absence of glycerol. Therefore, the blood ascofuranone concentration after administration of the preventive or therapeutic agent according to this embodiment is preferably 0.0019 nM or more, more preferably 0.033 nM or more, even more preferably 0.1 nM or more, 0.33 nM or more, and most preferably 1 nM or more, 3.3 nM or more. The number of administrations may be once or multiple times per day, or once every few days.

When the preventive or therapeutic agent according to this embodiment contains glycerol, the amount of glycerol to be administered can be determined as appropriate. In this embodiment, by using ascofuranone-producing bacteria instead of an ascofuranone purified product as the active ingredient, it is expected that the amount of glycerol to be administered can be reduced compared to when an ascofuranone purified product is used.

The preventive or therapeutic agent according to this embodiment can be used by being contained in medicines, food and beverages, feed, etc.

[Pharmaceuticals]
The pharmaceutical product according to this embodiment contains, as an active ingredient, a preventive or therapeutic agent for a disease caused by the parasite according to this embodiment.

The additives, carriers, and administration methods (route, dose, and number of times) for the pharmaceuticals of this embodiment are the same as those described for the preventive or therapeutic agents of this embodiment.

The pharmaceuticals according to this embodiment also include products that are sold as quasi-drugs and have preventive or therapeutic effects against diseases caused by parasites.

The pharmaceutical product according to this embodiment may contain, as an active ingredient, a preventive or therapeutic agent for a disease caused by the parasite according to this embodiment in combination with a known preventive or therapeutic agent for a disease caused by a parasite. Examples of known preventive or therapeutic agents include suramin, pentamidine, melarsoprol, eflornithine, quinapyramine, and isometamidium.

[Food and drink]
The food and drink according to this embodiment contains a preventive or therapeutic agent for a disease caused by the parasite according to this embodiment.

Food and drink products are not particularly limited, but preferred examples of application include foods for specified health uses, foods with functional claims, and other health foods. For example, the preventive or therapeutic agent can be obtained as a mixture by mixing the above-mentioned preventive or therapeutic agent with an existing food and drink product. Specific examples of liquid, gel, powder, or solid food and drink products include water, tea, milk, soft drinks, fruit drinks, soups, miso soup, yogurt, jellies, jams, desserts, seasonings, noodles, processed foods, and dairy products. The products can also be in the form of tablets, capsules, powders, granules, pellets, jellies, drinks, etc.

The additives and additives for food and beverages and the administration method (route, dose, number of times) for the food and beverages according to this embodiment are the same as those explained for the preventive or therapeutic agent according to this embodiment, but the administration route is oral administration.

〔feed〕
The feed according to this embodiment contains a preventive or therapeutic agent for a disease caused by the parasite according to this embodiment.

The feed may be, but is not limited to, feed for mammals (livestock) such as cattle, horses, pigs, sheep, goats, and deer, and feed for birds (poultry) such as chickens and quails. For example, the preventive or therapeutic agent may be added to existing feed as a feed additive to obtain a compounded product.

The feed additives and administration method (route, dose, number of times) for the feed of this embodiment are the same as those explained for the preventive or therapeutic agent of this embodiment, but the administration route is oral.

Pharmaceutical Compositions
The pharmaceutical composition according to this embodiment contains ascofuranone-producing bacteria containing ascofuranone produced by the bacteria itself as an active ingredient.

The definitions of each term, ingredients other than the active ingredients (additives or carriers), content, administration method (route, dose, number of times), etc., in the pharmaceutical composition of this embodiment are the same as those explained in the section on the preventive or therapeutic agent for diseases caused by parasites in this embodiment.

The pharmaceutical composition according to this embodiment can be used as a medicine or can be contained in a medicine. It is particularly useful as a medicine for preventing or treating diseases caused by parasites.

[Effects of this embodiment]
In this embodiment, since the active ingredient contains ascofuranone-producing bacteria, which have been shown to have a higher preventive or therapeutic effect against diseases caused by parasites than ascofuranone purified products, the amount of ascofuranone administered and/or the amount of glycerol administered can be reduced compared to when ascofuranone purified products are used.

In addition, in this embodiment, ascofuranone-producing bacteria that have not been subjected to extraction and purification treatment are used, the recovery rate of ascofuranone produced by the bacteria can be increased compared to the case of using a purified ascofuranone product that has been subjected to extraction and purification treatment, thereby improving the productivity of a preventive or therapeutic agent for diseases caused by parasites.

The present invention will now be described with reference to examples, but the present invention is not limited to these examples.

Example 1: Evaluation of preventive and therapeutic effects of oral administration of purified ascofuranone product and dried ascofuranone-producing bacterial cells against Trypanosoma evans infection Trypanosoma evans IL1695 strain was used as the trypanosome (represented as protozoa in Table 1) and mice (Balb/c mice, female, 9 weeks old, weighing approximately 20 g) were infected intraperitoneally with 1 x 10 ⁴ cells/head. Ascofuranone purified product (shown as "purified product" in Table 1) or ascofuranone-containing dried ascofuranone-producing bacteria (shown as "producing bacteria" in Table 1) was mixed with PBS solvent containing various concentrations of glycerol (0, 40, 75 w/w%) to prepare test solutions of various concentrations (PBS solvent only in Test Example 19), and 200 uL per day was orally administered to mice (n=1 in each Test Example) using an oral probe. Oral administration was performed for a total of 14 days, including the day of infection with trypanosoma, 6 days before infection, and 7 days after infection, with the day being considered as day 0.

For 30 days after infection, trypanosomes appearing in the blood were observed under a microscope using a hemocytometer, and the blood trypanosome concentration in each test example was quantitatively evaluated (lower limit of detection: 10 ⁵ cells/mL). In addition, the survival and death of mice were confirmed for 30 days after infection. The results are shown in Table 1.

The ascofuranone-producing bacterial dry material used was an ascofuranone-producing bacterial cell (Acremonium sclerotigenum F-1392 strain ascG-disrupted strain in which the ascI gene was highly expressed (ΔascG-I strain)) that had been cultured by the method described in the aforementioned Patent Document 4 (WO 2018/207928) to produce at least 1 g/L (culture solution) of ascofuranone, sterilized (heated at 121°C for 30 minutes), recovered (ascofuranone recovery rate: >80%), and dried by dry heating.

On the other hand, the purified ascofuranone used was the same as that used in Patent Document 2 (Patent Publication No. 4468638).

Mice not administered ascofuranone (Test Example 19) developed parasitemia and died 10 days after infection. All mice administered ascofuranone purified product alone (without glycerol) (Test Examples 1-3) and mice administered ascofuranone-producing dried bacteria alone (without glycerol) (Test Example 10) survived, but trypanosoma infection was temporarily observed in the blood. On the other hand, mice administered ascofuranone purified product and glycerol simultaneously (Test Examples 4-9) and mice administered ascofuranone-producing dried bacteria and glycerol simultaneously (Test Example 13) did not show trypanosoma infection in the blood and survived until the end of the observation period. This shows that the addition of glycerol improves the preventive and/or therapeutic effect of trypanosoma infection by oral administration of ascofuranone. This is expected to have the effect of reducing the oral dose of ascofuranone.

Moreover, mice administered only the purified ascofuranone product (Test Examples 2-3) showed temporary trypanosoma infection in the blood, whereas mice administered only the dried ascofuranone-producing bacteria cell product containing the same amount of ascofuranone (Test Examples 11-12) showed no trypanosoma infection in the blood. This shows that the dried ascofuranone-producing bacteria cell product is more effective in preventing and/or treating trypanosoma infection when administered orally than the purified ascofuranone product. This is expected to have the effect of reducing the oral dosage of ascofuranone and/or glycerol.

Example 2: Evaluation of preventive and therapeutic effects of oral administration of purified ascofuranone product and dried ascofuranone-producing bacterial cells against Trypanosoma congolense infection Trypanosoma congolense IL3000 strain was used as the trypanosome (represented as protozoa in Table 2), and mice (Balb/c mice, female, 9 weeks old, weighing approximately 20 g) were infected intraperitoneally with 1 x 10 ⁴ cells/head. A purified ascofuranone product (shown as "purified product" in Table 2) or a dried ascofuranone-producing bacterial cell containing ascofuranone (shown as "producing bacteria" in Table 2) was mixed with PBS solvent containing various concentrations of glycerol (0, 40, 75 w/w%) to prepare test solutions of various concentrations (PBS solvent only in Test Example 15), and 200 uL per day was orally administered to mice (n=1 in each Test Example) using an oral probe. Oral administration was performed for a total of 14 days, including the day of infection with trypanosoma, 6 days before the infection, and 7 days after the infection, with the day being considered as day 0.

The evaluation method was the same as in Example 1. The results are shown in Table 2. However, in Example 2, the blood trypanosome concentration on the day of death was not evaluated.

The dried ascofuranone-producing bacteria and purified ascofuranone product used were the same as those used in Example 1.

Mice that were not administered ascofuranone (Test Example 15) developed parasitemia and died on the 7th day after infection. Groups administered ascofuranone purified product alone (without glycerol) (Test Examples 1 and 2) developed parasitemia, and mice administered 20 mg/kg of ascofuranone purified product (Test Example 1) died on the 8th day after infection, and mice administered 30 mg/kg of ascofuranone purified product (Test Example 2) died on the 18th day after infection.

Among the groups in which purified ascofuranone and glycerol were administered simultaneously (Test Examples 3 to 5), mice administered 10 mg/kg of purified ascofuranone (Test Example 3) developed parasitemia and died 21 days after infection. On the other hand, mice administered 20 mg/kg of purified ascofuranone (Test Example 4) and 30 mg/kg (Test Example 5) showed no trypanosoma infection in the blood and survived until the end of the observation period.

Among the groups (Test Examples 6 to 8) administered only the dried ascofuranone-producing bacteria (no glycerol administration), mice administered the dried ascofuranone-producing bacteria (10 mg/kg ascofuranone equivalent) (Test Example 6) and mice administered the dried ascofuranone-producing bacteria (20 mg/kg ascofuranone equivalent) (Test Example 7) developed parasitemia, with the former dying on the 9th day after infection and the latter dying at the end of the 30-day observation period. On the other hand, mice administered the dried ascofuranone-producing bacteria (30 mg/kg ascofuranone equivalent) (Test Example 8) showed no trypanosoma infection in the blood and survived until the end of the observation period.

In the groups in which the dried ascofuranone-producing bacteria and glycerol were administered simultaneously (Test Examples 9 to 14), no trypanosoma infection was found in the blood of any of the mice, and they survived until the end of the observation period.

Comparing the results of Test Examples 1 and 7, 2 and 8, and 3 and 9, it is clear that the dried ascofuranone-producing fungus cell product has a higher preventive and/or therapeutic effect on trypanosoma infection when administered orally than the purified ascofuranone product. This is expected to reduce the oral dosage of ascofuranone and/or glycerol.

The above results show that the dried ascofuranone-producing cells are more effective when administered orally than the purified ascofuranone product, but this is thought to be because the coexistence of the ascofuranone-producing cells components with ascofuranone increases the solubility of ascofuranone in the mouse body, and/or it is gently digested and slowly transferred into the blood, and/or the amount excreted without being transferred into the blood is reduced, thereby increasing the blood concentration and retention time of ascofuranone. Using the dried ascofuranone-producing cells as an antiparasitic agent has the effect of reducing the amount of ascofuranone or glycerol administered compared to when a purified ascofuranone product is used, and furthermore, the ascofuranone recovery rate during preparation from the dried ascofuranone-producing cells is high at >80%, making it extremely useful industrially.

Claims

A preventive or therapeutic agent for diseases caused by parasites, containing ascofuranone-producing bacteria containing ascofuranone produced by the bacteria itself as an active ingredient.
The preventive or therapeutic agent according to claim 1, wherein the parasite is Trypanosoma congolense.
The preventive or therapeutic agent according to claim 1, wherein the parasite is Trypanosoma evansi.
The preventive or therapeutic agent according to claim 1, wherein the disease is trypanosomiasis.
The preventive or therapeutic agent according to claim 1, wherein the ascofuranone-producing microorganism is a microorganism of the genus Acremonium.
The preventive or therapeutic agent according to claim 1, wherein the ascofuranone-producing bacteria is a bacteria that does not produce ascochlorin.
The preventive or therapeutic agent according to claim 1, which contains glycerol.
The preventive or therapeutic agent according to claim 1, which is for oral administration.
A pharmaceutical comprising a preventive or therapeutic agent according to any one of claims 1 to 8.
A food or drink containing the preventive or therapeutic agent according to any one of claims 1 to 8.
A feed containing the preventive or therapeutic agent according to any one of claims 1 to 8.
A pharmaceutical composition containing ascofuranone-producing bacteria as an active ingredient, which contains ascofuranone produced by the bacteria itself.