JP2002212079A - Endotoxin inhibitor containing diacylglycerol derivative as active ingredient - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new endotoxin inhibitor, to provide a preventive/curative medicine for inflammation caused by the endotoxin, to provide an anti-septicemic medicine, and to provide food and drink or cosmetics containing the above medicines. SOLUTION: This endotoxin inhibitor contains a 3-O-[α-D- glycopyranosyl-(1-4)-α-D-glycopyranuronosyl]-1,2-diacylglycerol derivative as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an endotoxin inhibitor, a prophylactic / therapeutic agent for inflammation induced by endotoxin, and an antiseptic agent containing a diacylglycerol derivative having a specific structure as an active ingredient. The present invention also relates to a food or drink or cosmetic containing the above-mentioned drug. Further, the present invention relates to a method for efficiently producing the diacylglycerol derivative.

[0002]

2. Description of the Related Art Endotoxin (lipopolysaccharide), a component of the cell wall of Gram-negative bacteria, is a major component that interacts with living organisms in Gram-negative bacterial infections. The immune system, which is very sensitive to endotoxin and pathological phenomena caused by endotoxin in humans and animals, is strongly activated. That is, when endotoxin enters the blood, systemic inflammatory response syndrome (SIRS) ranging from endotoxin shock to sepsis due to the immune response induced by endotoxin
Is triggered. It is known that such endotoxin-induced sepsis is caused by endotoxin acting on various cells such as monocytes to produce inflammatory mediators (Advances in Immunology (Advances in Immunology)). Immunology) 53 volumes,
267-289, 1993).

As inflammatory mediators, interleukin 1, interleukin 6, interleukin 8, TNF-α and the like are important. These inflammatory mediators play a central role in the regulation of biological functions in the inflammatory response and defense mechanisms against infection (Annual Reviews of Biochemistry (AnnualR)
eviews of Biochemistry) 59, 783-836, 1990). In addition, endotoxin causes B
It induces a very wide range of immune responses, including nonspecific proliferation of cells and elevation of membrane proteins such as ICAM-1 expressed on macrophages and dendritic cells. Sepsis is a serious and fatal infectious complication that causes death, including irreversible hypotension and multiple organ failure (The Lancet)
338, 732-736, 1991). In particular, endotoxin is known to be a major mediator of sepsis due to Gram-negative bacteria. The symptoms of sepsis are reproduced in various animals by injecting endotoxin, and treatment to remove endotoxin from the blood is often given to sepsis patients, but complicated treatment such as the need to circulate blood once outside the body Is the law.
Although a method of reducing the shock response by using a neutralizing antibody against endotoxin has also been adopted, there is a problem that the protein factor is rapidly degraded in the blood,
At present, these methods are still insufficient.
Despite several treatments for gram-negative sepsis, high mortality is still a major clinical problem. A drug that suppresses an immune response or an inflammatory response caused by endotoxin (hereinafter, referred to as “endotoxin inhibitor”) is expected to be used as a prophylactic / therapeutic agent for endotoxin-induced inflammation and an antiseptic agent. For this reason, endotoxin inhibitors having excellent medicinal effects are desired in medical sites and the like.

[0004]

SUMMARY OF THE INVENTION The present invention has been made in view of the above problems, and has as its main object to provide a novel endotoxin inhibitor, a prophylactic / therapeutic agent for inflammation induced by endotoxin, and an antiseptic agent. .

[0005]

Means for Solving the Problems The present inventors have conducted intensive studies to search for novel endotoxin inhibitors using the inhibitory activity of endotoxin on TNF-α production from macrophages as an index. As a result, they have found that a specific glycerol derivative has an activity of inhibiting TNF-α production by endotoxin, and that it is useful as an antisepsis agent, thereby completing the present invention. In addition, the present inventors have found a method for efficiently producing the glycerol derivative in the course of research. That is, the present invention relates to the following objects and methods. 1. An endotoxin inhibitor comprising a compound represented by the general formula (I) or a derivative thereof as an active ingredient.

[0006]

Embedded image (In the formula, R1 or R2 is one selected from lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, tridecanoic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, oleic acid, linoleic acid, and linolenic acid. ) 2. An agent for preventing or treating endotoxin-induced inflammation, comprising a compound represented by the above general formula (I) or a derivative thereof as an active ingredient. 3. An antiseptic agent comprising a compound represented by the above general formula (I) or a derivative thereof as an active ingredient. 4. The above 1. ~ 3. A food, beverage, or cosmetic, comprising the drug according to any one of the above. 5. Culturing a microorganism of the genus Brevundimonas, and collecting the compound of the general formula (I) from the culture,
A method for producing the compound according to claim 1.

[0007]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below. 1. Regarding the compound represented by the general formula (I) The endotoxin inhibitor, the prophylactic / therapeutic agent for inflammation induced by endotoxin or the antiseptic agent of the present invention is a compound represented by the following general formula (I) (hereinafter referred to as "formula (I )) Or a derivative thereof as an active ingredient.

General formula (I):

Embedded image (In the formula, R1 or R2 is one selected from lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, tridecanoic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, oleic acid, linoleic acid, and linolenic acid. ) The compound of formula (I) has an α at position 3 of the diacylglycerol.
-D-glucopyranosyl- (1-4)-A type of glycerol derivative containing an α-D-glucopyranuronosyl group. Also,
The compound is structurally 3-O- [α-D-glucopyranosyl- (1-4)
-α-D-glucopyranuronosyl] -1,2, -diacyl-glycerol derivative. In the compounds of formula (I), R1 or R2 may be various fatty acids. Fatty acids include, for example, lauric acid, myristic acid,
Palmitic acid, stearic acid, arachidic acid, tridecanoic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, oleic acid, linoleic acid, and linolenic acid. As a more specific structure of the compound of the formula (I), a compound represented by the following general formula (II) (hereinafter, referred to as “compound of the formula (II)”) is exemplified.

General formula (II):

Embedded image Compound of formula (II) (1-O-asclepoyl-2-O-palmitoyl-3-O
-[α-D-glucopyranosyl- (1-4) -α-D-glucopyranuronosy
l] glycerol) corresponds to a compound of the formula (I) wherein R1 is an isomer of palmitic acid and R2 is an isomer of oleic acid. Compound of formula (II) is Caulobactor bacteroides NP-1
05 is known as a membrane component (Biochimica et Biophysica Acta)
1347, 127-139, 1997).

As described in the examples, the compounds of formula (II) can be purified from cultures of the genus Brevundimonas and have pharmacological activity as endotoxin inhibitors and antiseptic agents. The compound of formula (I) is a derivative whose structure is partially modified or modified as long as it has an effect as an endotoxin inhibitor, a prophylactic or therapeutic agent for inflammation induced by endotoxin or an antiseptic agent. You may. In the present invention, the hydroxyl group at the 3-position of the glucose residue or glucuronic acid residue of the formula (I) may be methylated, ethylated, propylated, isopropylated, acetylated or sulfated. Further, the hydroxyl group at the 2-position of a glucose residue or a glucuronic acid residue may be an amino group. Further, the hydroxyl group bonded to the carbonyl group at the 6-position of the glucuronic acid residue may be an ester such as a methyl group, an ethyl group, a propyl group or isopropyl. Examples of the derivative of the compound of the formula (I) include pharmacologically acceptable salts, esters and prodrugs. The pharmacologically acceptable salt is not particularly limited, but includes, for example, alkali metals (such as sodium and potassium) and alkaline earth metals (such as magnesium and calcium). These hydroxides or carbonates, alkali metal alkoxides (sodium methoxide, potassium t-
And the like. Examples of the salt include an acid addition salt to which an inorganic acid (hydrochloric acid, sulfuric acid, phosphoric acid) or an organic acid (maleic acid, citric acid, fumaric acid, etc.) is added, an amine addition salt, an amino acid addition salt, and the like. No.
In addition, the hydrate of the above-mentioned salt is also included in the salt mentioned here.

The ester is not particularly limited as long as it is an ester formed by an esterification reaction with an alcohol or a carboxylic acid. Examples of the alcohol include methanol, ethanol, 1-propanol, and 2-propanol. Examples of the carboxylic acid include formic acid, acetic acid, and lactic acid.

The term "prodrug" means a compound which is converted into the compound of the formula (I) or a derivative thereof after being administered to a living body and exerts the above-mentioned medicinal effects. Prodrug-modified compounds for the purpose of improving drug stability and absorbability, reducing side effects, and the like are also included in the derivatives of the present invention.

2. Process for Producing Compound of Formula (I) In the present invention, the compound of formula (I) or a derivative thereof may be produced by any method,
A microorganism capable of producing the compound, for example, Brevundimona
A method of purifying from a microorganism culture of the genus s, Caulobactor, or Streptomyces, a chemical synthesis method, a semisynthesis method, and the like can be widely used. In a method for purifying the compound of the formula (I) from a microorganism culture, it is important to efficiently purify the target compound from a crudely purified product containing the microorganism culture. The step of extracting with an organic solvent is useful as an efficient purification method. In particular, acetone is preferable as the organic solvent.
Acetone may be used alone or may contain up to 50% water. It is also possible to use a water-soluble organic solvent such as methanol, ethanol, propanol and butanol instead of acetone. A water-insoluble organic solvent such as ethyl acetate is important for efficiently concentrating the compound of the formula (I) from the crude product extracted with the water-soluble organic solvent. The compound of the formula (I) can also be directly extracted from a crudely purified microorganism culture using a water-insoluble organic solvent. Also, a pharmacologically acceptable salt which is a derivative of the compound of the formula (I) is obtained by adding an alkali metal, an alkaline earth metal, an alkali metal alkoxide to the compound of the formula (I).
It can be produced by reacting an inorganic acid or an organic acid. Further, the ester can be converted to a compound of formula (I) in the presence of an acid catalyst.
Can be produced by reacting an alcohol or a carboxylic acid with the compound of the formula (1). The formula (I) used in the present invention
May be purified to a single compound, or may be a crude product in the middle of the purification step.

3. Use of the agent of the present invention The endotoxin inhibitor of the present invention, a prophylactic / therapeutic agent for inflammation induced by endotoxin or an antiseptic agent (hereinafter, referred to as
The usage of the “genetic agent of the present invention” will be described. The medicament of the present invention comprises a compound of the formula (I) as an active ingredient and suppresses an endotoxin-induced immune or inflammatory response, thereby exerting its efficacy as a prophylactic / therapeutic agent for endotoxin-induced inflammation and an antiseptic agent. Demonstrate. The agent of the present invention is medically useful, for example, as a preventive or therapeutic agent for endotoxin shock, lethality, fever, inflammation, and Shwartzman reaction induced by contamination of blood with endotoxin. The drug of the present invention is also scientifically useful as a reagent for research on endotoxin and sepsis, and the like. The medicament of the present invention can be used as a medicament, food or drink, or cosmetics, as it is, or as an additive to these products. The medicament of the present invention may be used alone as a medicine, food or drink, or cosmetic, or may be used in combination with other endotoxin inhibitors, anti-inflammatory agents or anti-sepsis agents. The specific usage will be described below.

(1) Drug The drug of the present invention comprises a compound of the formula (I) as an active ingredient,
It can be used as a pharmaceutical as it is or by formulating it with a known pharmaceutical carrier. The agent of the present invention
For example, they can be formulated as oral preparations such as tablets, granules, powders and syrups, and parenteral preparations such as suppositories and external preparations. The pharmaceutical carrier is not particularly limited and includes, for example, solid carriers (starch, lactose, carboxymethyl cellulose, etc.),
Examples thereof include liquid carriers (distilled water, physiological saline, aqueous glucose solution, ethanol, propylene glycol, etc.) and oily carriers (various animal and vegetable oils, white petrolatum, paraffin, etc.). The medicament can be used for humans and non-human animals (pets, livestock). The dose of the above-mentioned medicine may be appropriately set according to the symptoms, sex, and age of the patient or the like who uses the medicine. For example, an adult may take the compound of formula (I), which is an active ingredient, in a daily dose of about 0.1 to 5000 mg per adult.

(2) Foods and drinks By adding the drug of the present invention to foods and drinks, the foods and drinks can be provided with an endotoxin inhibitory activity, a prophylactic / therapeutic activity for endotoxin-induced inflammation or an antiseptic activity. . Foods and drinks to be added are not particularly limited, but include meat products, processed vegetables, prepared foods, dairy products, confectionery, bread, soft drinks, fruit drinks, alcoholic beverages, and the like.
The compounding ratio of the anti-inflammatory agent of the present invention to foods is not particularly limited, either, but the compound of formula (I), which is an active ingredient, may be added so that about 0.1 to 5000 mg per adult per day can be taken.

Further, the anti-inflammatory agent of the present invention and other food materials may be mixed and formed into granules, powders, tablets, blocks, or the like to obtain food materials or health foods. Other food materials include, for example, sugars, edible proteins, alcohol, vitamins, thickening polysaccharides, amino acids,
Calcium salts, pigments, fragrances, preservatives and the like.

(3) Cosmetics By adding the agent of the present invention to cosmetics, the cosmetics can be provided with an endotoxin inhibitory activity, a prophylactic / therapeutic activity for inflammation induced by endotoxin or an antiseptic activity. Cosmetics are not particularly limited,
For example, lotions, cosmetic creams, emulsions, foundations, lipsticks, hair styling products, hair tonics, hair growth products, toothpastes, mouthwashes, shampoos, rinses and the like. When preparing cosmetics, oils and fats such as vegetable oils, waxes such as lanolin and beeswax, hydrocarbons, fatty acids, higher alcohols, various surfactants, pigments, fragrances, vitamins, plant and animal extract components And those commonly used as raw materials for cosmetics, such as ultraviolet absorbers, antioxidants, preservatives, etc., can be produced as appropriate.

The compounding ratio of the drug of the present invention to cosmetics is not particularly limited either, but the compound of the formula (I), which is an active ingredient, is percutaneously absorbed in an amount of about 0.1 to 5000 mg per adult per day. It may be added at an appropriate ratio.

[0020]

Examples Examples of the method for producing the compound of the formula (II), the endotoxin inhibitory activity test, the antiseptic activity test and the production of pharmaceuticals, foods and drinks and cosmetics containing the compound of the formula (II) are shown below.

Example 1 Method for producing compound of formula (II): A compound of formula (II) was purified as an endotoxin inhibitor from a microorganism culture of the genus Brevundimonas. The endotoxin inhibitory activity of the fraction obtained at each purification step is as follows:
It was measured by the method described in Example 2. First, Brevundimon
as sp. Y381 (Deposit No .: NISL-B-4725
Noda Institute of Scientific Research) was replaced with 12 L of medium (glucose 2%, starch 1%, Meat Extract 0.1%, dry yeast 0.4%, soy flour 2.5%) sterilized by an autoclave.
%, Salt 0.2%, K ₂ HPO ₄ 0.005%, pH 7.3)
Cultured for one day. The obtained cells were collected and extracted with 66% acetone overnight at 4 ° C. After concentration of the acetone extract under reduced pressure,
The pH was adjusted to 2 with hydrochloric acid, the soluble fraction was extracted with ethyl acetate, and concentrated under reduced pressure to obtain 15.4 g of a solid. After resolubilizing the solid in ethyl acetate, the solid was applied to a silica gel column, and hexane, toluene, toluene: ethyl acetate (1: 1),
When elution was performed in the order of ethyl acetate, ethyl acetate: acetone (1: 1), acetone, acetone: methanol (1: 1), and methanol, two kinds of eluted fractions (F5 and F5) were obtained.
7) was obtained. F5 had endotoxin inhibitory activity, and was eluted with ethyl acetate: acetone (1: 1) to obtain 2.82 g of a crude product. F7 was eluted with acetone: methanol (1: 1) to obtain 2.86 g of a crude product. As a result of analysis of the purified sample, F7 was found to be structurally similar to the compound of F5, but did not show endotoxin inhibitory activity. F5 was further eluted using silica gel chromatography with ethyl acetate: MeOH: water solvent.
Eluted by changing the composition of [16: 2: 1, 8: 2: 1, 4: 2: 1]
487.8 mg of an active fraction was obtained. Chromatography was performed using Sephadex LH20 equilibrated with chloroform: methanol (1: 1) to obtain 372.0 mg of an active fraction. The active fraction was adsorbed on a DEAE-Toyopearl column equilibrated with chloroform: methanol: water (30: 60: 8) to remove unadsorbed substances, and then chloroform: methanol: 0.8M sodium acetate (30: 60: 8). To obtain 262.8 mg of F5 as a final purified sample. F7 is obtained by adsorbing an active fraction on a DEAE-Toyopearl column equilibrated with chloroform: methanol: water (30: 60: 8) to remove non-adsorbed substances.
Chloroform: methanol: 0.8M sodium acetate (3
0: 60: 8) to obtain 228.6 mg of an eluted fraction.
Ethyl acetate: MeO using silica gel chromatography
H: The solvent of water was eluted while changing the composition to [12: 2: 1, 4: 2: 1] to obtain 37.0 mg of F7 as a final purified sample.

The final purified preparations F5 and F7 were converted to silica gel TL.
It was subjected to C, HPLC, IR, NMR, and MS, and structural analysis was performed. F5
1 and 4, the ¹ H-NMR spectrum is shown in FIGS. 2 and 5, and the ¹³ C-NMR spectrum is shown in FIGS. 3 and 6. In addition, the fatty acid side chain was determined by methanolysis of the purified sample to convert it to fatty acid methyl ester, followed by GC and GC-MS to compare with the standard product.
In addition, asclepoyl group (or cis-11,12-methyleneoctadeca
Confirmation of the position of the double bond of the noyl group) was determined by fragmentation using a tandem-MS method. As a result of the structure analysis, F5 having endotoxin inhibitory activity was found to be a compound represented by the following general formula (II).

Formula (II):

Embedded image F7 having no endotoxin inhibitory activity is a compound represented by the following general formula (III) (1-O-asclepoyl-2-O-pa
lmitoyl-3-O-α-D- (6'-sulfo) quinovopyranosylglycero
l) was found to be.

General formula (III):

Embedded image This compound has the same fatty acid side chain as Formula (II), but differs in the substituent at the 3-position of glycerol. This difference is considered to affect the presence or absence of endotoxin inhibitory activity.

Example 2 Measurement of Endotoxin Inhibitory Activity by TNF-α Production Inhibition Test: Cells produce TNF-α by the action of endotoxin, which induces an inflammatory response. It can be said that the higher the TNF-α production suppression rate, the stronger the endotoxin inhibitory activity. The endotoxin inhibitory activity of the compound of formula (II) was determined by measuring the effect of the compound of formula (II) on the amount of TNF-α produced by the cells.
First, mouse macrophage tumor cells J774A.1 were transformed into RPMI-
1640 (10% FBS, 50 mM HEPES, 0.1 mM 2-mercaptoethanol,
100 unit / ml penicillin, 100μg / ml streptomycin)
The cells were grown in a medium and suspended in an RPMI medium to a final concentration of 2.5 × 10 ⁵ cells / ml. Sigma endotoxin was added to a final concentration of [2 μg / ml]. On the other hand, after dissolving the compound of formula (II) obtained in Example 1 in methanol, R
It was diluted to a final concentration of 50-0.1 μg / ml with PMI medium and added to the previously prepared J774A.1 cell suspension. After culturing overnight in a CO ₂ incubator, the culture supernatant was collected and TN produced.
The amount of F-α was measured by ELISA (Pharmingen Opt.
EIAkit). The TNF-α production inhibition rate (%) when the compound of formula (II) was added to the test system at various concentrations was determined by the following formula. Table 1 shows the results. Formula: 100 × (1-A / B) (A is the amount of TNF-α production when J774A.1 cells are stimulated with endotoxin in the presence of the compound of formula (II). B
Is the amount of TNF-α production when J774A.1 cells are stimulated with endotoxin in the absence of the compound of formula (II). )

[0026]

[Table 1]

As is evident from Table 1, the compound of formula (II) showed a concentration-dependent increase in TNF-α caused by endotoxin.
Production was suppressed. Compounds of formula (II) have been shown to be useful as endotoxin inhibitors and as prophylactic and therapeutic agents for endotoxin-induced inflammation.

Example 3 Measurement of Endotoxin Inhibitory Activity by B Cell Growth Inhibition Test: B cells start to proliferate by the action of endotoxin and are often used as an indicator of endotoxin activity. The higher the B cell proliferation inhibition rate,
It can be said that endotoxin inhibitory activity is strong. By measuring the effect of the compound of formula (II) on B cell proliferation,
The endotoxin inhibitory activity of the compound of formula (II) was measured. Spleen cells were prepared from spleens of BALB / c mice according to a conventional method, suspended in RPMI-medium at a final concentration of 5 × 10 ⁵ cells / ml, and plated on a 96-well plate. Sigma endotoxin was added to a final concentration [5 or 2 μg / ml], and the compound of formula (II) obtained in Example 1 was dissolved in methanol, and then dissolved in RPMI medium to a final concentration of 100 to 0.1 μg / ml. The resulting mixture was diluted to a ml and added to the previously prepared spleen cell suspension. 7
After culturing in a CO ₂ incubator for 2 hours, 0.5 μCi / ml of tritiated thymidine was pulsed for the last 4 hours. After the cells were collected on a glass filter using a cell harvester, tritiated thymidine incorporated into the proliferated B cells was measured with a beta scintillation counter to serve as an indicator of endotoxin-induced B cell proliferation. The B cell growth inhibition rate (%) when the compound of formula (II) was added to the test system at various concentrations was determined by the following formula. Table 2 shows the results. Calculation formula: 100 × (1-A / B) (A is the amount of tritiated thymidine taken up by B cells grown by endotoxin stimulation in the presence of the compound of formula (II) [DP
M]. B is the amount of tritiated thymidine taken up by endotoxin-stimulated B cells in the absence of the compound of formula (II)
[DPM])

[0029]

[Table 2]

As is clear from Table 2, the compound of formula (II) inhibited endotoxin-induced B cell proliferation in a concentration-dependent manner. Compounds of formula (II) have been shown to be useful as endotoxin inhibitors and as prophylactic and therapeutic agents for endotoxin-induced inflammation.

[Example 4] Measurement of anti-sepsis activity using mouse sepsis model: The effect of the compound of formula (II) on mouse sepsis was evaluated by the following method. <Animals used>: ICR female mice weighing 25 to 30 g (6 to 8 weeks old). <Experimental method>: The compound of the formula (II) obtained in Example 1 was replaced with 0.2
% Tween-20 in physiological saline, and the suspension was intraperitoneally administered to four mice in each group at a rate of 100 mg / kg body weight, and after 5 minutes, 40 mg of endotoxin was intraperitoneally administered. . When the survival rate after 24 hours was evaluated, it was 0% in the control group to which only the physiological saline was administered.
Was 100% in the group that had been treated with the compound in advance. Furthermore, the substituent of the glyceryl at the 3-position is different from the formula (III)
In the group similarly using the compound (F7 of Example 1), 25
%Met. From the above results, it was shown that the compound of the formula (II) has a remarkable antisepsis effect even at a biological level, and is useful as an antisepsis agent.

[Embodiment 5] Cytotoxicity test The compound of formula (II) obtained in Example 1 was subjected to a cytotoxicity test by the following method. RPMI-1640 (10% FBS, 50 mM HEP
ES, 0.1mM 2-mercaptoethanol, 100 unit / ml penicilli
n, 100 μg / ml streptomycin) medium in a tumor cell line (J
774A.1 and PU5-1R) and normal cell lines (FDC-P2 and CTLL-2;
WEH containing interleukin (IL) -3 as each growth factor
The supernatant after I-3 cell proliferation and IL-2 from Pharmingen were added). In addition, DMEM (10% FBS, 50 mM HEPES,
0.1mM 2-mercaptoethanol, 100 unit / ml penicillin,
100 μg / ml streptomycin)
Endothelial cells collected from LB / 3T3 and bovine aorta were grown. 5-10 × 10 ³ cells of each were treated with 100 μl of the compound of formula (II).
The cells were cultured in the presence of g / ml and evaluated by counting the number of cells after 3 to 5 days. The compound of formula (II) did not show cytotoxicity in any of the cells.

2. Single dose toxicity test: The compound of formula (II) was subjected to a single dose toxicity test by the following method. <Animals used>: ICR female mice weighing 25 to 30 g (6 to 8 weeks old). <Experimental method>: According to the guidelines for toxicity test methods for pharmaceuticals (February 15, 1984, Pharmaceutical Affairs Commission No. 118, notification from the Ministry of Health and Welfare Pharmaceutical Affairs Bureau second section manager to prefectural health administration director)
A single dose toxicity study was performed. The compound of formula (II) is suspended in a physiological saline containing 0.2% Tween-20, and the suspension is added to 5 mice per kg of body weight.
It was administered intraperitoneally at a rate of 100 mg and observed for 14 days. As a result, no animal died in all cases, and no apparent side effect was observed. Therefore, the compound of the formula (II) was found to be extremely low in toxicity.

[Example 6] Drug containing the drug of the present invention: Formulation of the compound of the formula (II) obtained in Example 1 by the following method to prevent and treat endotoxin inhibitor, endotoxin-induced inflammation. Drug that can be used as a drug or an antiseptic agent. (1) The same amount of lactose and 5 g of magnesium stearate were mixed with 100 g of the compound of the formula (II), and the mixture was tableted with a single-shot tableting machine to produce tablets having a diameter of 10 mm and a weight of 300 mg. (2) The tablets obtained in (1) were pulverized, sized and sieved to obtain granules of 20 to 50 mesh.

Example 7 Food and drink containing the drug of the present invention: (1) As a food containing the drug of the present invention, a candy having the following composition (parts by weight) was produced. Sugar (47.
0), syrup (49.76), fragrance (1.0), water (2.
0), a compound of the formula (II) (0.24). (2) As a beverage containing the agent of the present invention, orange juice having the following composition (parts by weight) was produced. Commercially available orange juice (99.76), compound of formula (II) (0.2
4).

Example 8 Cosmetics Containing the Agent of the Present Invention (1) A toothpaste having the following composition (parts by weight) was produced. Dicalcium phosphate (42), glycerin (18),
Carrageenan (0.9), sodium lauryl sulfate (1.
2), sodium saccharin (0.09), butyl paraoxybenzoate (0.005), compound of formula (II) (0.0
5), perfume (1) and water were added to give 100.

(2) A lotion having the following composition (parts by weight) was prepared. Lyserin (5.0), propylene glycol (4.0), compound of formula (II) (0.15), polyoxyethylene sorbitan monolaurate (2.
0), ethanol (10.0), fragrance (0.1), and purified water were added to give 100.

(3) A hair rinse having the following composition (parts by weight) was produced. Stearyl dimethyl (1.4), benzylammonium stearyl alcohol (0.6), glycerin monostearate (1.5), salt (0.1), compound of formula (II) (0.1) and purified water In addition, it was set to 100.

[0039]

Industrial Applicability According to the present invention, there are provided an endotoxin inhibitor, a prophylactic / therapeutic agent for inflammation induced by endotoxin and an antiseptic agent comprising a compound of the formula (I) as an active ingredient. The agent of the present invention is particularly medically useful as an agent for preventing or treating endotoxin shock, lethality, fever, inflammation, and Shwartzman reaction induced by contamination of blood with endotoxin derived from Gram-negative bacteria. The drug of the present invention is also scientifically useful as a reagent for research on endotoxin and sepsis, and the like. The medicament of the present invention can be used as a medicament, food or drink, or cosmetics, as it is, or as an additive to these products. The medicament of the present invention may be added alone to pharmaceuticals, foods and drinks or cosmetics, or may be used in combination with other endotoxin inhibitors, anti-inflammatory agents or anti-sepsis agents.

[Brief description of the drawings]

FIG. 1 is an IR spectrum of the endotoxin inhibitory activity fraction (F5) obtained in Example 1.

FIG. 2 is a ¹ H-NMR spectrum of an endotoxin-inhibiting activity fraction (F5) obtained in Example 1.

FIG. 3 is a ¹³ C-NMR spectrum of the endotoxin inhibitory activity fraction (F5) obtained in Example 1.

FIG. 4 is an IR spectrum of the fraction (F7) obtained in Example 1.

FIG. 5 is a ¹ H-NMR spectrum of the fraction (F7) obtained in Example 1.

FIG. 6 is a ¹³ C-NMR spectrum of the fraction (F7) obtained in Example 1.

Continuation of the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (reference) A61P 29/00 A61P 29/00 4C083 31/04 31/04 4C086 C12P 19/44 C12P 19/44 4C087 // A61K 7 / 00 A61K 7/00 M 7/08 7/08 7/16 7/16 C07H 15/04 C07H 15/04 D (C12P 19/44 (C12P 19/44 C12R 1:01) C12R 1:01) (72) Inventor Riichiro Uchida 250 Noda, Noda, Chiba Prefecture Kikkoman Co., Ltd. (72) Inventor Goo Sato 250 Noda, Noda, Chiba Prefecture Kikkoman Co., Ltd. (72) Riki Masuda, Noda, Chiba No. 399 Noda F-term in Noda Institute of Industrial Science (Reference) 4B014 GB06 GB07 GG07 GK05 GL03 4B017 LC03 LE10 LG02 LK11 4B018 LB01 LB08 MD27 MD85 ME14 MF01 MF13 4B064 AF41 BE09 BE12 BE14 BH02 BH04 BH11 DA02 CE07 CC01 DD01 JJ10 4C083 AB292 AC102 AC122 AC352 AC422 AC442 AC482 AC692 AC782 AC862 AD352 AD391 AD392 BB48 CC04 CC39 CC41 DD22 DD23 EE11 EE33 4C086 AA01 EA05 GA17 MA01 MA04 MA16 MA28 MA57 MA63 NA14 ZA67 ZA90 ZA92 ZB11 ZC37 4C087 BC22 BC31 CA10 MA16 MA28 MA57 MA63 NA14 ZA67 ZA90 ZA92 ZB11 Z

Claims (5)

[Claims] 1. An endotoxin inhibitor comprising a compound represented by the following general formula (I) or a derivative thereof as an active ingredient. General formula (I): (In the formula, R1 or R2 is one selected from lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, tridecanoic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, oleic acid, linoleic acid, and linolenic acid. ) 2. A preventive or therapeutic agent for inflammation induced by endotoxin, comprising a compound represented by the following general formula (I) or a derivative thereof as an active ingredient. General formula (I): (In the formula, R1 or R2 is one selected from lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, tridecanoic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, oleic acid, linoleic acid, and linolenic acid. ) 3. An antiseptic agent comprising a compound represented by the following general formula (I) or a derivative thereof as an active ingredient. General formula (I): (In the formula, R1 or R2 is one selected from lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, tridecanoic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, oleic acid, linoleic acid, and linolenic acid. ) (4) A food, drink, or cosmetic product containing the drug according to any one of (1) to (3). 5. The method for producing a compound according to claim 1, wherein the microorganism of the genus Brevundimonas is cultured, and the compound of the following general formula (I) is collected from the culture. General formula (I): (In the formula, R1 or R2 is one selected from lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, tridecanoic acid, pentadecanoic acid, heptadecanoic acid, nonadecanoic acid, oleic acid, linoleic acid, and linolenic acid. )