JPH0338530A - Antifungal agent - Google Patents

Abstract

PURPOSE:To obtain an antifungal agent containing a fatty acid and hydrolytic enzyme and having low toxicity for animal and non-animal and high activity. CONSTITUTION:The antifungal agent containing 5-22C straight- and branched- chain saturated and unsaturated fatty acid and hydroxyfatty acid, especially 10-12C saturated fatty acid and unsaturated fatty acid of 10-20C monotriene and hydrolytic enzyme (e.g. protease, lipase, beta-1,3-glucanase, chitinase, phospholipase or cellulase). The antifungal agent can be used by adding it as antifungal agent or mildewproofing agent to cleaning agent, cosmetic, etc., in addition to medicinal use for man and other animal, enhances drug effects and lowers the dose. The side effects can be reduced by using together with a conventional antifungal agent.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to antifungal agents.

[Conventional technology]

Many drugs have been developed for, for example, mycoses caused by various fungi, and some are in clinical use. For example, imidazole drugs such as miconazole and bifonazole, polyene drugs such as nyscutin and amphotericin B, thiocarbamate drugs such as triazole, triazole drugs, griseofulvin,
There is too much time to list such things as virolnidoline and potassium iodide.

However, these drugs almost always have some side effects, causing symptoms such as fever, chills, headache, gastrointestinal disorders, kidney disorders, and liver disorders. In addition, fatty acid-based undecylenic acid and its salts have low toxicity, but their antifungal action is relatively weak.

In addition, attempts have been made to suppress the growth of fungi using lytic enzymes (mainly containing hydrolytic enzymes) derived from microorganisms such as actinomycetes, but none have been found to have sufficient efficacy.

Furthermore, in order to increase the activity of bacteriolytic enzymes, a method of coexisting fatty acids has been reported, but this is for bacteria (Physiol, Plant, 27).
, 187, 194 (1972). ) It was not known for fungi that have different cell surface structures.

[Problem to be solved by the invention]

The object of the present invention is to develop antifungal agents with low toxicity and high activity for animals and non-animals, and in particular to develop agents with low toxicity and antifungal action for humans and other animals.

[Means to solve the problem]

The present inventor discovered that when various fatty acids and hydrolyzed oxygen such as protease were mixed and allowed to act on fungi, a bacteriolytic phenomenon accompanied by a change in the morphology of fungal cells was observed. For example, when fatty acids and hydrolytic enzymes are added to a yeast cell, which is a type of fungus, the cell wall maintains a relatively normal morphology, but the protoplasm contracts and cell contents leak out of the cell. , a unique bacteriolytic phenomenon never seen before was observed. These phenomena are caused by fatty acids causing some damage to the surface layer of fungal cells,
Furthermore, it is presumed that bacteriolysis progresses synergistically by the action of hydrolytic enzymes on the cell surface layer, leading to the completion of the present invention.

That is, the present invention relates to an antifungal agent containing a fatty acid and a hydrolytic enzyme. According to the present invention, it can be used for both veterinary and non-veterinary purposes.

Here, the fatty acid used in the present invention has 5 to 5 carbon atoms.
22, it may be any of straight chain and branched saturated fatty acids, straight chain and branched unsaturated fatty acids, and hydroxy fatty acids. In the case of saturated fatty acids, carbon atoms of 10 to 15 are particularly preferred. Examples include capric acid, undecylic acid, lauric acid, tridecanoic acid, myristic acid, incapric acid, isolauric acid, isomyristic acid, and isopentadecanoic acid.

Furthermore, in the case of unsaturated fatty acids, monoenes, dienes, and trienes having 10 to 20 carbon atoms are particularly preferred. For example, undecylic acid, myristoleic acid, palmitoleic acid,
These include oleic acid, linoleic acid, and lylunic acid.

Further, the hydrolase used in the present invention is not particularly limited, but protease, lipase, β-1,3-glucanase, chitinase, phospholipase, and cellulase are more preferable. The protease may be trypsin, pepsin, papain, chymotrypsin, subillysin, serrapeptase, promelain, streptonachise, pronase, etc., and its origin does not matter.

The antifungal agent used in the present invention may contain at least one of each of the fatty acids and hydrolytic enzymes described above,
You can freely combine them.

In addition, in addition to the fatty acids and hydrolytic enzymes mentioned above, conventionally used antibacterial agents and bactericidal agents may be used in combination, and
It is also possible to add absorption aids, solubilizers, etc. for these drugs. For example, by coexisting a fatty acid and a hydrolase with existing antifungal agents such as amphotericin B and griseofulvin, the efficacy of the antifungal agent can be increased, and as a result, the dosage of these agents can be lowered and their side effects can be reduced.

The antifungal agent of the present invention can be used as an antibacterial agent in detergents, cosmetics, etc. in addition to pharmaceutical applications for humans and other animals.

It can also be added and used as a fungicide. For example, by adding fatty acids and hydrolytic enzymes to laundry detergents, it is possible to remove protein-derived stains and sterilize mold and the like.

The dosage form of the antifungal agent of the present invention used for medical purposes can also be, for example, tablets, powders, capsules, solutions, sugar coatings, and the like.

Specific drugs that can be mixed into tablets, capsules, etc. are shown below. Binders such as tragacanth, gum arabic, corn starch or gelatin;
Excipients such as microcrystalline cellulose; leavening agents such as corn starch, pregelatinized starch, alginic acid, etc.;
lubricant Niji 914, such as magnesium stearate;
sweetening agents such as lactose or saccharin; flavoring agents such as peppermint, red radish oil or cherry; and when the dosage unit form is a capsule, materials of the above type may further contain a liquid carrier such as an oil or fat. can. Various other materials may be present as coatings and to otherwise modify the physical form of the dosage unit. For example, tablets may be coated with shellac, sugar, or both. A syrup or elixir may contain the active compound, Ci41 as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor.

Sterile compositions for injection include the active substance dissolved or suspended in a vehicle such as water for injection, a naturally occurring vegetable oil such as sesame oil, coconut oil, peanut oil, cottonseed oil and the like or a synthetic fatty vehicle such as ethyl oleate and the like. It can be formulated according to normal formulation practices. Buffers, preservatives, antioxidants, etc. can be combined as necessary.

The antifungal agents created in this way include Rhodotorula spp., Candida spp., Curbutococcus spp., Trichosporon spp., Sassocharomyces spp., Geotrichum spp., Schizosafucalomyces spp.
Coccidioides, Histoplasma, Plastomyces, Mucor, Aspergillus, Rhizopus,
Effective against many fungi such as Trichophyton.

When the antifungal agent of the present invention is used as a medicine, its administration route may be either oral or parenteral. The dose is determined by the patient's age, weight, medical condition and method of administration. Usually, the daily dose is 0.0 for oral administration.
1 to 2000 μ/kg, and 0.1 to 2000 μ/kg for parenteral administration.
01 to 1000 ■/mutsu.

Furthermore, the toxicity of the antifungal agent of the present invention is clearly lower than that of commonly used imidazole drugs, polyene drugs, and the like. The degree of toxicity is the same as the simple sum of the toxicity of fatty acids and hydrolytic enzymes alone. For example, in the case of lauric acid and trypsin of the present invention, the toxicity is low (
Each LD.

(mouse intravenous injection> 131■/kg, 89■/kg), amphotericin B (L Dso (mouse intravenous injection) 4mg
/kir).

The present invention will be specifically explained with reference to Examples.

〔Example〕

Example 1 An antibacterial test was conducted on the antifungal agent of the present invention. The test bacteria were yeast of the genus Rhodotorula (Rhodotorula glutinis).
otorula glutinis IFO075
4), various fatty acids listed in Table 1 (final concentration 0.1
After adding 0.0 ml and various enzymes (final concentration 1 ml/all) to the yeast suspension and reacting at 30° C. for 4 hours, changes in the morphology of the yeast cells were observed under a phase contrast microscope.

Here, one loopful of the test bacteria was mixed with YM medium (medium composition: peptone 5 g/1, yeast extract 3 g/l).

Malt extract 3g/lc or more, Difco product), glucose l Og/1. After inoculating at pH 6,5), 2
Shaking culture was carried out at 5°C for 16 hours. After centrifugal collection, the bacterial concentration is about 10? -10'' cells/ml, and this was used as the yeast test solution.

In addition, add 0.5 ml of fatty acid to each test tube in advance, add 4.5 rsl of yeast test solution to each test tube, and finally add 0.5 sj of enzyme solution! was added to a predetermined concentration to start the reaction.

As shown in Table 1, no bacteriolysis was observed when fatty acids or enzymes were used alone, but yeast cells were rapidly lysed when enzymes and fatty acids coexisted.

Moreover, among fatty acids, lauric acid, myristoleic acid, isopentadecanoic acid, etc. were particularly effective.

No. table In addition, Each symbol in Table 1 means the following.

+: Against more than 90% of bacteria. its contents (original Leakage of traits (traits) was observed.

+: Leakage of contents was observed from about 20-90% of bacteria.

±: Leakage of the contents was observed in 20% or less of the bacteria.

: No change in the appearance of the bacteria was observed.

In addition, the number of viable bacteria in the specimen in which +activity was observed decreased to 1/20 to 1150 compared to before the reaction.

Example 2 The antibacterial activity of fungi other than the genus Dotorula was investigated in the same manner as in Example 1, and the results are shown in Table 2.

Here, as a fungus, Candida albicans (IFO
1060), Cryptococcus albidus (IFO0)
610), Sanki Romyces cerevisiae (CBS
1171),) Lycosboro cutaneum (IFO01)
16), Aspergillus niger (IFO6341)
-Used.

Enzymes such as trypsin and pronase were prepared at a rate of 1/lan.

In addition, the fatty acids are 0.1■/l1 for lauric acid, myristoleic acid, and isopentadecanoic acid, respectively.
1 was added to the guiding yarn, but both showed the same results. It was also found to be effective against pathogenic bacteria such as canchia infection (caused by Candida genus) and cryptococcosis (caused by Cryptococcus genus).

Table 2 4, Effects of the Invention The antifungal agent of the present invention is expected to be put to practical use as a drug that is effective and has few side effects, for example, in the treatment of fungal diseases. Therefore,
The present invention is extremely useful in the pharmaceutical industry.

Claims (3)

[Claims] (1) Antifungal agent containing fatty acids and hydrolytic enzymes. (2) Claim in which the number of carbon atoms in the fatty acid is in the range of 5 to 22 (
1) The antifungal agent described above. (3) Hydrolyzed oxygen causes protease, lipase, β-1
, 3-glucanase, chitinase, phospholipase and cellulase.
1) The antifungal agent described above.