JP2003183179A - Endotoxin blood treating agent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain an inhibitory or protective agent against a bioreaction caused by an endotoxin in an organism without the necessity for using an expensive adsorption column. <P>SOLUTION: This endotoxin blood disease treating agent contains a compound having a neuraminidase inhibitory action as the effective component. The compound having the neuraminidase inhibitory action is (-) ethyl (3R,4R,5S)-4- acetamide-5-amino-3-(-1-ethyl-proboxy)cyclohexa-1-en-1-carboxylate monophosphate or [(+)-(4S,5R,6R)-5-acetyl-amino-4-guanidino]-6-[(1R-2R)-1,2,3- trihydroxy-propyl]-5,6-dihydro-4H-pyrane-2-carboxylic acid hydrate. The agent is useful also for curing endotoxin-induced sepsis. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a therapeutic agent for endotoxemia. Specifically, it relates to a novel drug that suppresses or protects a biological reaction caused by endotoxin.
The therapeutic agent for endotoxemia of the present invention is also useful as a therapeutic agent for sepsis caused by endotoxin.

[0002]

BACKGROUND OF THE INVENTION Endotoxin is a lipopolysaccharide (lipopolysaccharide) in the cell wall of Gram-negative bacteria.
Hereinafter also referred to as "LPS". ) Is a component of endotoxin. Biological reactions due to this endotoxin include fever, inflammation, changes in hemodynamics (increased heart rate, hypercoagulability, etc.), shock, paralytic ileus, and multiple organ failure. In order to suppress or prevent these symptoms, a mechanism for removing endotoxin from the living body or inhibiting expression of endotoxin action in the living body is necessary.

Conventionally, as a method for removing endotoxin from a living body, blood adsorption therapy using an endotoxin-removing fiber (polymyxin B-immobilized fiber) is known. However, this therapy is expensive because the adsorption column is expensive, the insurance coverage is limited to two uses, and one adsorption column is used for about two hours each time, so it is used continuously. There is a problem that you cannot do it.

[0004]

In view of the above situation, it is an object of the present invention to provide an inhibitor or antifungal agent for a biological reaction caused by endotoxin in a living body, which does not require the use of an expensive adsorption column. And

The present inventors have realized that drug therapy as a causative therapy is necessary to remove endotoxin from the living body or to inhibit its action, on the basis of grasping the problems of the blood adsorption therapy. Therefore, the present invention is aimed at. After many tests, we reached the involvement of sialic acid dissociation, which is considered to be one of the pathogenic mechanisms of endotoxemia, and that neuraminidase inhibitors that suppress this sialic acid dissociation are effective for the treatment of endotoxemia. Thought. As a result of further research, the present invention was finally completed. Further, the therapeutic agent for endotoxemia according to the present invention is developed for the purpose of both treatment of endotoxemia and treatment of sepsis caused by endotoxin. The therapeutic agent for endotoxemia according to the present invention can completely solve the above problems.

[0006] Regarding the development of the present invention, a reference document was "Galanos, C. et al: Galactosamine-induc.
ed sensitization to the lethal effects of endotoxi
n, Proc.Natl.Acad.Sci. USA, 76 (11), 5939-5943,1979 ''
And the "Treatment of rabbits, rats, and
mice with D-galactosamine increased their sensitiv
ity to the lethal effects of lipopolysaccharide se
The description "veral thaus and fold." was particularly useful. Also, `` Okamura, T. et al: Effect of neuraminid
ase on anaphylactic contraction of guinea pig taen
ia coli (1) .Jpn.J.Allergol., 29 (12) .1028-1034,1980 ''
See also “After the pretreatment wi
th neuraminidase, the contraction of smooth muscle
provoked by the antigen added completely disappear
ed. ”was useful. Furthermore, "Yasuo Suzuki:
Influenza virus and receptor sugar chain recognition, protein, nucleic acid, enzyme, 43 (16), 2559-2566, 1998 ”are also helpful. Especially,“ influenza A and B viruses are common regardless of hemagglutinin subtype. Sialyl lactotype I {Neu
5Acα2-6 (3) Gal β1-3Glc NAcβ1-} and II {Neu5Acα2
-6 (3) Gal β1-4Glc NAc β1-} type sugar chain is strongly recognized. Was useful.

[0007]

In order to solve the above-mentioned problems, the invention described in claim 1 of the present invention is a therapeutic agent for endotoxemia containing a compound having a neuraminidase inhibitory activity as an active ingredient.

The invention according to claim 2 of the present invention is the same as the invention according to claim 1, wherein the compound having a neuraminidase inhibitory action is oseltamivir phosphate ((-)-ethyl (3R, 4R, 5S ) -4-Acetamido-5-amino-3
It is a therapeutic agent for endotoxemia which is-(-1-ethylpropoxy) cyclohex-1-ene-1-carboxylate monophosphate.

Further, in the invention described in claim 3 of the present invention, in the invention described in claim 1, the compound having a neuraminidase inhibitory action is zanamivir hydrate ((+)-
(4S, 5R, 6R) -5-Acetylamino-4-guanidino-6-[(1R,
2R) -1,2,3-Trihydroxypropyl] -5,6-dihydro-4H-
Pyran-2-carboxylic acid hydrate) is a therapeutic agent for endotoxemia.

Further, in the invention described in claim 4 of the present invention, the therapeutic agent for endotoxinemia is useful as a therapeutic agent for sepsis caused by endotoxin.
The therapeutic agent according to any one of 1. Hereinafter, the present invention will be described in more detail.

The present invention is a therapeutic agent for endotoxemia, which comprises a compound having a neuraminidase inhibitory action as an active ingredient. In the present invention, the neuraminidase inhibitory action means an action of inhibiting the enzyme activity of neuraminidase which dissociates sialic acid from the sugar chain terminal of the cell membrane. Examples of the compound having a neuraminidase inhibitory action include oseltamivir phosphate, zanamivir hydrate and Neuramic acid derivatives such as Neu5Ac2en. Among them, oseltamivir phosphate is particularly useful. Oseltamivir phosphate is one of the neuraminidase inhibitors, and its composition is (-)-ethyl (3R, 4R, 5S) -4-acetamido-5-amino.
It is represented by -3- (1-ethylpropoxy) cyclohex-1-ene-1-carboxylate monophosphate. Oseltamivir phosphate is conventionally known as an anti-influenza virus agent.

Zanamivir hydrate is also useful as a compound having a neuraminidase inhibitory action. Zanamivir hydrate is one of the neuraminidase inhibitors and its composition is (+)-(4S, 56,6R) -5-acetylamino-4
-Guanidino-6-[(1R, 2R) -1,2,3-trihydroxypropyl] -5,6-dihydro-4H-pyran-2-carboxylic acid hydrate. Zanamivir hydrate is conventionally known as an anti-influenza virus agent.

The drug containing the compound having a neuraminidase inhibitory activity of the present invention as an active ingredient is extremely useful as a therapeutic agent for endotoxemia and also as a therapeutic agent for endotoxin-induced sepsis.

In the present invention, the dose of oseltamivir phosphate or zanamivir hydrate is based on the amount used as an anti-influenza virus agent. Regarding usage, oseltamivir phosphate follows the usage as an anti-influenza virus agent, but zanamivir hydrate is dissolved in physiological saline and administered subcutaneously or intravenously.

In the present invention, the reason why these compounds having a neuraminidase inhibitory action can suppress or prevent the biological reaction caused by endotoxin is that endotoxin is involved in the activation of neuraminidase. This is well demonstrated pharmacologically by the following test examples. Hereinafter, the present invention will be described in more detail with reference to test examples.

[0016]

[Test Example 1] <Confirmation test for therapeutic effect of oseltamivir phosphate on endotoxemia> (1) Test method Hartley male guinea pig (350 to 450 g) was pentobarbita
l The anesthesia was opened under anesthesia, and a minimal force transducer (5 × 3 mm, F-041S, Star medical. Co.) was attached to the colon by a suture. Next, a cylindrical transmitter (12 mm diameter x 36 m long) connected to this transducer with a cable.
m, 10T-F) was implanted subcutaneously on the back of the guinea pig. Radio waves from the transmitter are received by the antenna inside the receiver and output.
It was recorded by connecting to a PC with A / D. The post-operative guinea pigs were allowed to drink and eat freely in a cage, and 4 days later, LPS (lipopolysaccharide, Escher ) was dissolved in physiological saline.
ichia coli , serotype O111: B4) was intraperitoneally administered at 0.3 mg / kg to induce endotoxemia. Oseltamivir phosphate 100 mg / kg was dissolved in physiological saline and orally administered 1 hour before LPS administration.

(2) Test Results FIG. 1 is a graph showing the effect of LPS and oseltamivir phosphate (hereinafter also referred to as “OT”) on the intestinal motility of guinea pig collateral. That is, guinea pig weight (k
g) physiological saline solution (2 ml) was intraperitoneally administered (2 ml / kg, ip)
This reaction was used as a control. Intestinal motility curves of guinea pigs when LPS was dissolved in this physiological saline solution, 0.3 mg / kg of LPS was intraperitoneally administered, and OT 100 mg / kg was dissolved orally administered in physiological saline 1 hour before LPS administration. Is shown in FIG. The vertical axis of FIG. 1 represents the intestinal tension (g) of the colon cord, and the horizontal axis represents the time (h). Figure 2
Affects LPS and O on intestinal tone of the guinea pig colon
It is a graph which shows the influence of T. The vertical axis in FIG. 2 represents the rate of change in tension when the intestinal tension (g) of the colon cord before LPS administration was set to 100% and the intestinal tension (g) after 2 hours of euthanasia in the guinea pig was set to 0%. . The horizontal axis indicates time (h).

The intestinal contractile movement of the guinea pig lacquer was recorded consciously and unrestrained for a long period of time by a telemetry system via a collateral-mounted transducer. In the guinea pigs to which LPS was administered, a characteristic suppression of intestinal motility was observed, which peaked 2 to 4 hours after administration. On the other hand, oseltamivir phosphate 100 mg / kg was orally administered 1 hour before LPS administration
In (p, o.) guinea pigs, LPS administration completely inhibited the suppressive action of intestinal tone characteristic of endotoxemia. Each test was conducted in 3 cases, and statistical treatment was performed. As a result, in the LPS administration group, 2 to 7 after LPS administration.
As compared with the control group by time, a statistically significant intestinal tone suppressing effect was exhibited, and as shown in FIG. 2, the oseltamivir phosphate pretreatment group recovered this intestinal tone suppressing effect to almost the same level as the control group.

(3) Findings on test results LPS administration in the intestinal contraction monitor of conscious / unrestrained guinea pig colonic cord is useful as an experimental model of human endotoxemia and endotoxin-induced sepsis by the present test. Was shown. In this experimental animal model, oseltamivir phosphate showed an effect of suppressing endotoxemia and endotoxin-induced sepsis. From this, oseltamivir phosphate can be applied as a therapeutic agent for endotoxemia and a therapeutic agent for sepsis caused by endotoxin.

[0020]

[Test Example 2] <Confirmation test for therapeutic effect of zanamivir hydrate on endotoxemia> (1) Test method In the same manner as in the method of Test Example 1, endotoxemia was induced in guinea pigs. Zanamivir hydrate 100 mg / kg was dissolved in physiological saline and subcutaneously administered immediately before LPS administration. Zanamivir hydrate (hereinafter also referred to as "ZN") was poorly absorbed by oral administration, and an increase in blood concentration could not be expected. Therefore, it was subcutaneously administered. Subcutaneous administration of zanamivir hydrate is LP
It was performed immediately before S administration.

(2) Test Results FIG. 3 is a graph showing the effects of LPS and ZN on the intestinal motility of guinea pig collateral. That is, 2 ml of physiological saline was intraperitoneally administered per guinea pig body weight (kg) (2 ml / k
g, ip), and this reaction was used as a control. A graph of the intestinal motility curve of the guinea pig when LPS was dissolved in this physiological saline, 0.3 mg / kg of LPS was intraperitoneally administered, and 100 mg / kg of ZN was subcutaneously administered immediately before the administration of LPS is shown in FIG. The vertical axis in FIG. 3 represents the intestinal tension (g) of the colon cord, and the horizontal axis represents the time (h). In guinea pigs subcutaneously (sc) administered with 100 mg / kg zanamivir hydrate (ZN) immediately before LPS administration, as shown in FIG. 3, the inhibitory effect of LPS administration on intestinal tone characteristic of endotoxemia is completely inhibited. Was done.

(3) Observations on test results In an experimental animal model of endotoxemia and endotoxin-induced sepsis, zanamivir hydrate showed an effect of suppressing endotoxemia and endotoxin-induced sepsis. From this, zanamivir hydrate can be applied as a therapeutic agent for endotoxemia and a therapeutic agent for sepsis caused by endotoxin.

[0023]

[Example 1] <Example 1 of production method of preparation for treatment of endotoxemia> Oseltamivir phosphate, which is a main raw material, was dissolved in physiological saline to make a total amount of 2 ml to prepare a preparation for oral administration.

[0024]

[Example 2] <Example 2 of production method of preparation for treatment of endotoxemia> 40 mg of zanamivir hydrate as a main raw material was dissolved in physiological saline to make a total volume of 2 ml to give a preparation for subcutaneous injection.

[0025]

INDUSTRIAL APPLICABILITY As described in detail above, the therapeutic agent for endotoxinemia according to the present invention enables a novel therapeutic method of drug treatment for endotoxemia. That is, use of the therapeutic agent for endotoxemia according to the present invention eliminates the need to rely on blood adsorption therapy and the use of an expensive adsorption column in the treatment of endotoxemia, and therefore its utility is extremely large. . Moreover, the therapeutic agent for endotoxemia of the present invention is also useful as a therapeutic agent for sepsis caused by endotoxin.

[Brief description of drawings]

FIG. 1 is a graph showing the effects of LPS and OT on the intestinal motility of the guinea pig colon cord in Test Example 1.

FIG. 2 is a graph showing the effects of LPS and OT on the intestinal tone of the guinea pig colonic cord in Test Example 1.

FIG. 3 is a graph showing the effects of LPS and ZN on the intestinal motility of the guinea pig colon cord in Test Example 2.

[Explanation of symbols]

LPS: Lipopolysaccharide OT: Oseltamivir phosphate ZN: Zanamivir hydrate

   ─────────────────────────────────────────────────── ─── Continued front page    (71) Applicant 501491273             Kamoto Nemoto             5-6-27 Higashifushimi, Nishi-Tokyo, Tokyo (72) Inventor Yasuhiro Suzuki             5-27-18 Nishishinkoiwa, Katsushika-ku, Tokyo (72) Inventor Tadao Okamura             8-12 Sakainan-cho, Musashino-shi, Tokyo Eight             Mansion 608 (72) Nobunbun Ninomiya             5-1-3 Sendagi, Bunkyo-ku, Tokyo (72) Inventor Kamoto Nemoto             5-6-27 Higashifushimi, Nishi-Tokyo, Tokyo F-term (reference) 4C062 BB29 BB37 BB40                 4C084 AA16 MA52 NA14 ZB352                 4C086 AA01 AA02 BA07 MA01 MA04                       MA52 NA14 ZB35                 4C206 AA01 AA02 GA30 MA01 MA02                       MA04 MA52 NA14 ZB35

Claims (4)

[Claims] 1. A therapeutic agent for endotoxemia, which comprises a compound having a neuraminidase inhibitory activity as an active ingredient. 2. A compound having a neuraminidase inhibitory activity is oseltamivir phosphate ((-)-ethyl (3R, 4R, 5S) -4-
The therapeutic agent for endotoxemia according to claim 1, which is acetamide-5-amino-3-(-1-ethylpropoxy) cyclohex-1-ene-1-carboxylate monophosphate). 3. A compound having a neuraminidase inhibitory activity is zanamivir hydrate ((+)-(4S, 5R, 6R) -5-acetylamino-4-guanidino-6-[(1R, 2R) -1, 2,3-trihydroxypropyl] -5,6-dihydro-4H-pyran-2-carboxylic acid hydrate), The therapeutic agent for endotoxemia according to claim 1. 4. The therapeutic agent according to any one of claims 1 to 3, which is useful as a therapeutic agent for endotoxin-induced sepsis.