WO1998035665A1

WO1998035665A1 - Counteractive for endotoxin

Info

Publication number: WO1998035665A1
Application number: PCT/JP1998/000598
Authority: WO
Inventors: Yoshiyuki Fujishima; Naoto Koyama; Hisao Ito; Kenzo Yokozeki; Masahiko Oshimura; Hisamine Kobayashi; Tadashi Takemoto; Toshihiko Ando; Satoko Azumi; Ken-Ichi Tanamoto
Original assignee: Ajinomoto Co., Inc.
Priority date: 1997-02-14
Filing date: 1998-02-13
Publication date: 1998-08-20

Abstract

An excellent counteractive for endotoxin, characterized by containing one or more fatty acids as active ingredients, and useful for preventing or treating septicemia and a local inflammation.

Description

Description Endotoxin neutralizer

(Technical field)

The present invention relates to an endotoxin neutralizing agent, and more particularly, to an endoxin neutralizing agent containing a fatty acid as an active ingredient.

The endotoxin-neutralizing component (fatty acids) of the present invention can be injected or administered into a vein, intraperitoneal cavity, breast, rumen (ruminal ruminal of a reproductive animal), or the like, and then be administered. A new endotoxin that inhibits the lethal effects of toxin, thus being effective in preventing and treating inflammatory diseases such as sepsis and mastitis in humans and animals due to Gram-negative bacteria It concerns the neutralizer.

(Background technology)

Endotoxin (lipopolysaccharide), known as a cell wall component of Gram-negative bacteria, is mainly composed of sugars and fatty acids. Phosphorylated diglucosamine, in which multiple long-chain fatty acids are amide- and ester-linked, is particularly called lipid A and is a common active unit that expresses various physiological actions of endotokins. ing.

Although the physiological actions of endotoxin are diverse and varied, it is well known that it produces lethal toxicity in susceptible animals, including humans. Endotoxin that has migrated into the blood by translocation of intestinal gram-negative bacteria, etc., is taken up by phagocytic cells such as macrophages when it is taken up by TNF and IL. It induces the production of inflammatory cytokines, such as eleven. As a result, a variety of eicanosides, such as vasodilator mediators such as nitric oxide, have been updated. As they are released into the blood one after another, they often trigger a systemic inflammatory response (sepsis). The resulting decrease in blood pressure due to dilatation of the peripheral vessels and changes in permeability impair organ microcirculation and often cause death due to multiple organ failure. On the other hand, in the case of local infections caused by gram-negative bacteria, for example, intramammary infections found in lactating cows, etc., endotoxin released into milk is converted to cytokines derived from locally present inflammatory cells. It stimulates production and often causes mastitis with a systemic inflammatory response. Endotoxin experimentally injected into the breast is known to cause various clinical symptoms of mastitis (swelling of the breast, fever, inability to stand, etc.). In addition, metabolic diseases such as abomasum displacement, laminitis, and hepatic disorder have frequently occurred in ruminants such as stomachs that have been heavily fed with concentrated feed. Various experiments have revealed that endotoxin is released from gram-negative bacteria in the rumen that died due to metabolic abnormalities in the rumen.

As the pathology of sepsis and mastitis due to microbial infections becomes apparent, how to control the systemic inflammatory response before shock or multi-organ failure, or confer resistance to infection, While this has been emphasized recently, the elimination and management of these triggering inflammatory reactions must, of course, be forgotten. Antimicrobial chemotherapy has been reported to be one of such attempts to remove or inactivate (neutralize) endotoxin in body fluids.

Measures to reduce endotoxin concentration in body fluids include reducing the influx of endotoxin from the intestinal tract into the blood and neutralizing endotoxin in the blood. It has been devised. In fact, a method of reducing endotoxin influx from the intestinal tract into the bloodstream by orally administering a substance capable of absorbing endotoxin (B. Ditteretal. , G as troentero , Vol. 84, 1547, 1983), polymixin complex (JP-A-3-2201998) and aliphatic amine (JP-A-5-271710). Methods for neutralizing endotoxin in the blood have been reported so far. In addition, reports of the application of a monoclonal antibody recognizing the lipid A moiety to the treatment of sepsis (EJ Ziegler et al., New England J. Med., 1991, 429) and polymixin B Attempts have been made to prevent shock in patients by fixing it to a Listen fiber and perfusing it with blood to remove endotoxin from the blood (M. Kodama et al., Endotoxin Research

Series, Vol. 1, 33, 1990). However, none of these methods can be expected to be clinically effective at present.

Endotoxin (lipid A) neutralizing antibody developed by Centa Co., Ltd., Zoma of Amerika has finally proved clinically effective despite extensive clinical trials. There was no. Also, in patients who need to neutralize endotoxin in the intestine or blood, it is often necessary to administer the neutralizing agent for a certain period of time, for example, until the intestinal inflammation subsides. There is. With this in mind, synthetic drugs such as those described above always carry the risk of side effects, and long-term administration is not always appropriate. On the other hand, a horseshoe crab-derived peptide (Japanese Patent Application Laid-Open No. 418280/1990) is known as an endotoxin neutralizing agent derived from a natural product, but it is necessary to supply a large amount of this peptide. However, it is hardly practical because of the inability to supply a large amount of the raw material, but also difficulty in easily refining the obtained crude peptides. In addition, Japanese Patent Publication No. 5-501416 discloses the prevention and treatment of the toxic effects of endotoxin by the combined use of iron-binding lactoferrin and IgG. A method for performing this is disclosed. This is thought to be mainly due to the bacteriostatic action of lactoferrin.

On the other hand, に関 is related to the removal and inactivation of endotoxins derived from pathogenic bacteria in milk. Thus, in practice, frequent milking is the only means and rarely uses such drugs for persistence, toxicity, or economics for such purposes. . Also, there have been very few such attempts. Regarding livestock metabolic disease, although there are attempts to prevent a decrease in pH in rumen, which is said to lead to the death of gram-negative bacteria in rumen, endotoxin in rumen fluid Very few attempts have been made to actively remove or inactivate these components.

Thus, a systemic or local inflammatory response, triggered by the killing of Gram-negative bacteria and the release of endotoxin in body fluids, often results in a shock (septic shock). High mortality rate, and the large impact on the quality and productivity of dairy products, etc., has become a major clinical and industrial problem. Nevertheless, there are still no good prophylactic and therapeutic agents found today.

The present invention is effective in the prevention and treatment of systemic inflammatory response syndrome (sepsis) in humans and animals caused by gram-negative bacteria and local inflammation such as mastitis and laminitis in the background of the prior art described in the preceding paragraph. The aim is to develop and supply new and superior endoxin neutralizers.

(Disclosure of the Invention)

The present inventors can inactivate endotoxin directly, or localize endotoxin in addition to the original endotoxin target cells (inflammatory cells) to achieve systemic activity. If the release of these cells from the media, which triggers the inflammatory response, could be suppressed, the survival rate of sepsis could be improved, or the livestock could be prevented or reduced by local inflammation. Being able to contribute to the improvement of productivity, we conducted intensive studies to find a neutralizing agent for endotoxin, and as a result, various fatty acids showed a neutralizing effect on endotoxin activity. Have Were newly found, and the present invention was completed based on such knowledge. That is, the present invention relates to an endotoxin neutralizer characterized by comprising one or more fatty acids as an active ingredient.

Hereinafter, the present invention will be described in detail.

Examples of the fatty acids which are the active ingredients of the endotoxin neutralizer of the present invention include free fatty acids, salts thereof and derivatives thereof.

Fatty acids are preferably fatty acids having 9 or more carbon atoms, more preferably saturated or unsaturated fatty acids having 9 to 22 carbon atoms. Are nonanoic acid, decanoic acid, lauric acid, myristic acid, myristoleic acid, palmitic acid, and phenol. Lumitoleic acid, oleic acid, linoleic acid, α-linolenic acid, r-linolenic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, etc. Can be mentioned. These fatty acids can be obtained by ordinary methods such as hydrolysis of natural fats and oils or isolation from natural products, and can be easily and inexpensively obtained. For fatty acids having a number of carbon atoms outside the above range, those with a smaller number of carbon atoms are less active, whereas those with a larger number of carbon atoms are difficult to solubilize in an aqueous solution and are therefore somewhat difficult to handle. .

Examples of the salt of the fatty acid include pharmaceutically acceptable salts thereof, for example, sodium salt, potassium salt, magnesium salt, calcium salt, ammonium salt and the like. it can.

Derivatives of fatty acids include derivatives that do not lose endotoxin neutralizing activity, such as amides and esters (monoglycerides, diglycerides, triglycerides). , Etc.), phospholipids (L-α-phosphatidylcolindilaurol, L_α-phosphatidylcolindimyristyl, L-α-phosphatidylcolindioleyl, soy lecithin, Egg yolk lecithin, etc.), glycolipids, protein-bound fatty acids, polyoxyethylene, Examples include steles, acid anhydrides, pyridine compounds, halides, sulfonates, sulfides, thiocyanates, etherates, acylates, azides, thiols, and the like.

One of these fatty acids may be used alone or two or more of them may be used in combination as an active ingredient of an endotoxin neutralizing agent. Can also.

The findings regarding the neutralizing action of endotoxin by the endotoxin neutralizing agent of the present invention are as follows. That is, first, the above-mentioned fatty acids used in the present invention are mixed with endotoxin in advance and kept at room temperature to inhibit the limulus reaction by endotoxin (test example). 1). In addition, the above fatty acids and endotoxin are mixed in advance, and then injected into mice whose sensitivity has been increased by administration of galactosamine, or by injection of endotoxin. Later administration of the above fatty acids suppresses the lethal effects of endotoxin injection, which would normally cause endotoxin shock and die (Test Examples 2 and 3) See). In addition, fatty acids, when ingested, do not undergo degradation in the gastrointestinal tract (including the stomach) from the oral cavity to the stomach, and reach the intestinal tract without being broken down. Can also be expected to act as an endotoxin neutralizer in the gastrointestinal tract. The endotoxin neutralizing agent of the present invention is administered to animals such as humans and livestock as a sterile injection solution intravenously, intraperitoneally, intramuscularly, in the breast, etc., or as an enteral infusion. You can do it. It can also be applied topically to the skin, eyes, ears, nose, anus, breast, vagina, etc. as a powder, solution, suspension, cream, ointment, spray, etc. In addition, they can be administered orally, for example, as powders, solutions, capsules, tablets, syrups, tinctures, etc., or directly as food or feed. The fatty acids are widely ingested as foods, and there is no problem in safety in oral administration. The invention of this invention In such an administration method, a doxoxine neutralizing agent can be administered in combination with an existing antibiotic.

In the preparation for administration, the preparation can be prepared by appropriately using excipients, stabilizers, and the like that are used in usual preparations. Also, known and suitable pharmacologically acceptable solvents (eg, water, ethanol, glycerol, propylene glycol, liquid polyethylene glycol, etc.), flavors, sweeteners, binders, isotonic Formulations can also be made with appropriate use of active substances, liniments, surfactants (such as bile acids), absorption delaying agents, PH regulators (such as triethanolamine), and the like.

In addition, the above-mentioned fatty acids may be used in addition to other components effective in alleviating the symptoms of endotoxin (eg, corticosteroids (methylprednisolone, dexamethasone, etc.), antibiotics, etc. (Ampicillin, methicillin, noncomicin, etc.) and antibodies (anti-endotoxin antibody, anti-interleukin-16 antibody, anti-TNF antibody, etc.) and the like. It is also possible to prepare an endotoxin neutralizing agent.

Prevention and treatment of local inflammation such as sepsis and mastitis in humans and animals by the endotoxin neutralizing agent of the present invention include, for example, direct injection, It can be mixed with food and drink in advance and administered orally.In the case of treatment, oral administration, injection, suppositories, and the application of a matrix containing fatty acids to mucous membranes can be used. It can be administered by control release.

The effective amount of the fatty acids for the prevention and treatment of local inflammation such as sepsis and mastitis is about 1 to 100 mg / kg of human or animal body weight per day. It is presumed that

(Best mode for carrying out the invention) Hereinafter, the present invention will be further described with reference to inspection examples. Inspection example 1

In this test example, fatty acids were preliminarily mixed with endotoxin, and the concentration of residual endotoxin after a certain period of time was determined by Limulus test. Then, the endotoxin neutralizing ability of those fatty acids was evaluated.

Lipopolysaccharide prepared from Salmonel la minnesota (S. minnesota) was used as the enodotocin (Wes tpha 1 et a 1 .. Z. Naturforsch., Vol. 76, 148-155, 1952). Fatty acids include radium, myristate, palmitate, oleic, linoleic, α-linolenic, r— Linoleic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid (all manufactured by Nacalai Tesque, reagent grade), L-α-phosphatidylco-lindilaurol, L_α — Those listed in Table 1 below were inspected, such as phosphatidylcolindi millistoil (above, manufactured by Sigma). That is, each of these fatty acids was dissolved in 99.5% ethanol to prepare a stock solution having a concentration of 20 mg Zml. The lipopolysaccharide was dissolved in distilled water for injection to give a solution having a concentration of 20 μg Zml. The evaluation by Limulustest was performed by diluting the above fatty acid stock solution with ethanol to a concentration of 10, 2.5, and 0.SSS mg Zml. Glycine sodium hydroxide buffer (0.2 M, pH 10.6 (1992 published by Hirokawa Shoten)

“Reagent preparation method”)) and endotoxin solution are added in a ratio of 1: 1: 2, and these mixtures are each pre-plated at room temperature for 1 hour and 30 minutes, and then added to distilled water for injection. LPS (Lipopolli Saccharide) is calculated

It was diluted to be around 100 pg / m1. Remaining LPS of diluted solution The activity was measured using a Limulus test reagent “Toxicolor System” (Seikagaku Corporation). The difference between the relative activity value of the reaction solution with the neutralizing substance and the control when the ra property was 100% when the same reaction was performed without fatty acids (control) and the neutralization ability was defined as neutralization ability. did. The test results are also shown in Table 1 below.

Table 1: Neutralizing effect of endotoxin by various fatty acids

(Endotoxin neutralizing ability of fatty acids (%))

Concentration (mg / ml)

: Elementary atom! Fatty acids 1 0 2.5 0.625

6 (0) hexanoic acid 0 0 0

8 (0) octanoic acid 0 0 0

9 (0) Nonanoic acid 1 5 6 0

10 (0) Decanoic acid 3 5 8 1 3

12 (0) Lauric acid 9 6 3 8 0

14 (0) myristic acid 7 8 9 6 7 8

16 (0) palmitic acid 6 2 9 1 7 1

14 (1) Myristoleic acid 9 3 5 2 9

16 (1) ', ° Lumitrain ^ 5 3 5 6 4 3

18 (1) Oleic acid 9 2 9 0 8 6

18 (2) Linoleic acid 9 6 9 1 6 2

18 (3) α — Linolenic acid 9 7 7 8 6 4

18 (3) r—linolenic acid 9 2 7 7 0

20 (4) Arachidonic acid 9 5 8 3 2 7

20 (5) Eicosanoic acid 9 600

22 (6) 9 7 9 4 5 4 1 fl—Phosphatic acid luciferic acid IH 8 5 7 7 8 1 1 phosphatic acid “lucorin acid” myristoyl 7 9 7 5 8 2

As is clear from Table 1, saturated and unsaturated fatty acids having 9 to 22 carbon atoms and phospholipids as derivatives thereof show inhibition of the Rimulus reaction. As a result, it was confirmed that the enzyme had an endotoxin neutralizing activity. Inspection example 2

In this test example, whether or not endotoxin was neutralized (in vitro) by fatty acids was evaluated by a mouse administration experiment.

Mice (C57BL / 6, female, Nippon Charles River Co., Ltd.) of 10 to 20 weeks of age were used for the administration experiment. The endoxin used was the same as in Inspection Example 1. Fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, α-linolenic acid, arachidonic acid, and docosahexane. The acid was checked. That is, each of these fatty acids was dissolved in 99.5% ethanol to give a stock solution having a concentration of 25 mg Zm1. The endotoxin was a solution having a concentration of 200 g Zm1. Take the fatty acid solution, sodium glycine hydroxide buffer (same as in Test Example 1) and edoxin solution at a ratio of 2: 3: 5, and use these in small test tubes. The mixture was mixed, ultrasonically suspended for 10 minutes, mixed and stirred for about 10 seconds, and then incubated at room temperature for 3 hours to obtain a sample. For the purpose of improving the sensitivity of LPS, the endotoxin concentration of this sample was adjusted to 0.00 in mice that had been injected intraperitoneally with 500 wi of galactosamine (concentration of 24 mg Zmi) in advance. It was diluted with distilled water for injection so that it became 1 to 10 μg Zm 1, and each was injected with 1001 and observed for life and death 18 hours later. The test results are shown in Table 2 below. Table 2: Neutralizing effect of endoxin by fatty acids

(Number of surviving mice Z Total number of mice administered)

Dosing endoxin (g)

Number of carbon atoms S ife) fatty acids 0.001 0.1

Ento ', toxin only 2/2 0 3

12 (0) Lauric acid 2 3 0 2

14 (0) myristic acid 1 2 2 2

16 (0) palmitic acid 5 5 2 2

18 (1) Oleic acid 5 5 2 2

18 (3) α-linolenic acid 3 3 1 2

20 (4) arachidonic acid 4 4 1 1

22 (6) Uto, Kosa Hekisan Rooster 2 2 0 1

As is evident from Table 2, 0.1 g of endotoxin alone is lethal, but 0.1 and 1 g of endotoxin were mixed with fatty acids (sample). Survival was also observed after administration of dotoxin, i.e., it was confirmed in animals that saturated and unsaturated fatty acids having 9 to 22 carbon atoms have a neutralizing effect on endotoxin. . Inspection example 3

In this test example, the possibility that endotoxin was neutralized in vivo by fatty acids was evaluated by a mouse administration experiment.

In this experiment, mice, endotoxin and fatty acids (but oleic acid) were all the same as those in Test Example 2. Endotoxin prepared a solution with a concentration of 200 μg Zml from this, Mix with the lactosamine solution so that the concentration of endotoxin is 0.2 μg Zm1 and the concentration of galactosamine is 24 mg Zml. It was adjusted. The oleic acid was dissolved in 99.5% ethanol to form a stock solution with a concentration of 25 mg Zm1, which was then added to a sodium glycine hydroxide buffer. (Same as in Test Example 1) were mixed in the ratios of 2: 3 and 1: 4. The above-mentioned galactosamine-endoxin solution (500-1) was intraperitoneally injected into mice, and immediately thereafter, the above-mentioned oleic acid-knock-off solution (100-1) was intraperitoneally injected. 18 hours after the intravenous injection, the cells were observed for life and death. The results of this observation are shown in Table 3 below.

Table 3: Neutralizing effect of endotoxin by fatty acids (in vivo) (survival number of mice and total number of administered mice)

Administered fatty acids (mg)

0 0 .5 1 .0 End toxin only 0/3

Entoxin + fatty acid 2 / z 3 4/5

As is evident from Table 3, endotoxin alone has a lethal dose of 0.1 g; it has the ability to separately produce 0.5 mg or 1.O mg of fatty acids (oleic acid). When administered, a clear life-saving effect was observed. Inspection example 4

In this test example, fatty acids or phospholipids were premixed with endotoxin in milk to determine the residual endotoxin content and activity in milk after a certain period of time. The endotoxin neutralizing ability of these fatty acids in milk was evaluated by measuring them in the test of administration to limulustest and mice, respectively.

The endotoxins were lipopolysaccharide (LPS) extracted from S. Minnesota and Escherichia coli (E. coil) 011 and E. col1 purchased from Sigma. LPS of i 0 128 was used. The free fatty acids are oleic acid (manufactured by Nakara Itesque), and the phospholipids are L-α-phosphatidylcholine dilauroyl and L-α-phosphatidylcholine. Myristoil, L-α-phosphatidylcolindioleoleyl (all manufactured by Sigma), egg yolk lecithin PC-98NJ (manufactured by Kyupie Co., Ltd.) and soybean lecithin (Tsurichi Lecithin Industrial ( Was inspected.

Oleic acid was dissolved in 99.5% ethanol and mixed with triethanolamine to form a solution having a concentration of 50 mg Zml. Also, oleic acid or phospholipids include sodium cholate (manufactured by Nakara Itesque), sodium taurocholate (manufactured by DFCO), and sodium deoxycholate. Mixed with sodium salt (manufactured by Nacala Itesque) or sodium salt of taurodeoxycholic acid (manufactured by Nacala Itesque), and diluted with Phosphate-buffered saline (PBS) (manufactured by Takara Shuzo). Similarly, a 50 mg Zml solution or emulsion was prepared. Dilute these solutions or emulsions with their respective blank solutions without neutralizing substances to give further concentrations of 12.5 mg / m3.lmg / ml and 0.8 mg Zml. A solution was prepared. LPS was used as a 10 μg / m 1 milk solution by mixing an aqueous solution having a concentration of l mg / ml with commercially available milk at a ratio of 1:99.

(a) The neutralizing substance solution (or emulsified liquid) and the LPS solution are mixed at 1: 1 and 1

: 2, 1: 4 or 1: 9 and kept at room temperature for 1 hour and 30 minutes Thereafter, the mixture was appropriately diluted with distilled water for injection, and the residual LPS activity was measured according to the method of Test Example 1. The neutralization rate was calculated from the relative activity value when the activity was 100% when the same reaction was performed in a blank solution excluding the neutralizing substance. The inspection results are shown in Table 4 below.

Table 4: Neutralizing Effect of Free Fatty Acids and Phospholipids on I-toxin in Milk Neutralizing Substance Buffer Popoli Final Concentration in Mixture (* 2) Maximum Limulus

Composition (* 1) S, y-Calide Neutralizing substance / LPS / Milk Neutralization rate (%) Oleic acid A S.m 1 nneso ta 0.25 / 5.0 / 50 100

0.26 ~ 16.6 / 6.7 / 67 100 0.16-10.0 / 8.0 / 80 62

B S. m i nneso ta 0.4-25 / 5.0 / 50 99

0.26-16.6 / 6.7 / 67 98 Phosphaticilcoli Β S.m 1 nneso ia 0.4-25 / 5.0 / 50 87 inch, 'Rawil 0.26-16.6 / 6.7 / 67 95

0.16-10.0 / 8.0 / 80 97

E. co 1 i 0111 0.4 ~ 25 / 5.0 / 50 94

0.26-16.6 / 6.7 / 67 94 0.16 ~ 10.0 / 8.0 / 80 92

E.col i 0128 0.4 ~ 25 / 5.0 / 50 89

0.26-16.6 / 6.7 / 67 91 0.16-10.0 / 8.0 / 80 73

C E. co 0111 0.26 ~ 16.6 / 6.7 / 67 100

0.16 ~ 10.0 / 8.0 / 80 89

D 0.26 ~ 16.6 / 6.7 / 67 100

0.16 ~ 10.0 / 8.0 / 80 93

Ε 0.26-16.6 / 6.7 / 67 100

0.16-10.0 / 8.0 / 80 98 0.8 ~ 5.0 / 9.0 / 90 60 Phosphatisi ,! Re B E.coli 0111 0.26-16.6 / 6.7 / 67 100 psi ', Myristil 0.16-10.0 / 8.0 / 80 89 Phosphatsi, I: B E.coli 0111 0.26 ~ 16.6 / 6.7 / 67 100 E.coli 0111 0.25 / 5.0 / 50 95

0.26-16.6 / 6.7 / 67 85 Soybean: / Chin B E.co 1 i 0111 0.4 ~ 25 / 5.0 / 50 97

0.26-16.6 / 6.7 / 67 91 氺 1: Knuffer composition;

A: 3% triethanolamine, 50% ethanol

B: 20mM sodium cholate, 5% 1ethanol, 80% PBS

C: 20 mM Na taurocholate, 5% ethanol, 80% PBS

D: 20mM Teseki Xicol Na, 5% ethanol, 80% PBS

E: 20mM Taurote "Shi Xicol ^ Na, 5% ethanol, 80% PBS Ne2 _: Unit of concentration;

Neutralizing substance: mg gm 1

L P S: u s / m I

milk : %

(b) Next, a solution of the neutralizing substance (L-α-phosphatidylcholine dilauroyl) and the LPS solution prepared as described above was injected into the peritoneal cavity of the mouse, and the LPS was actually injected. Was tested in vivo for inactivation. That is, 1001 was prepared by diluting the above mixed solution with distilled water for injection so that the LPS power became 1 gm1, and 500 mg of a galactosamine solution having a concentration of 24 mg / ml were dissolved in a mouse. Co-injected intraperitoneally simultaneously. The effect was determined according to the method of Test Example 2. The results of the test are as shown in Table 5 below.

5 Table: Neutralizing effect of toxin on phosphatidyl and ruchoyl "lauroyl in milk. Concentration in mixed solution. Mouse survival rate Neutralizing substance (mg / ml) LPS (/ ig / ml) Milk% (survival number / test All)

1 6 .6 6 .7 6 7 1 0 / '1 0

1 0 .0 8 .0 8 0 1 0 / '1 0

5 .0 9 .0 9 0 8 / '1 1

4.2.6.7.6 7 9 / '9

2.5 8 .0 8 0 9 / '9

0. 0 6. 7 6 7 4 No z 1 2

As is clear from this test example, when free fatty acids and phospholipids are used together with appropriate solubilizers and emulsifiers, endotoxin in milk can be effectively neutralized (inactivated). It turned out.

(Industrial applicability)

Fatty acids, which are the active ingredients of the neutralizing agent of the present invention, neutralize endotoxin and suppress the lethal effect of endotoxin, so that humans at the time of infection with Gram-negative bacteria can be used. And prevent harmful effects on the body due to local inflammation such as sepsis and mastitis in animals, and thus are effective in preventing and treating it. Further, an advantage of the present invention is that many such fatty acids are readily and inexpensively available. Therefore, according to the present invention, an excellent preventive and therapeutic agent for local inflammation such as sepsis and mastitis can be easily and inexpensively supplied.

Claims

The scope of the claims

1. An endotoxin neutralizer characterized by containing one or more fatty acids as an active ingredient.

2. The endotoxin neutralizing agent according to claim 1, wherein the fatty acid is a fatty acid having 9 or more carbon atoms.