WO1995003057A1

WO1995003057A1 - Endotoxin neutralizer

Info

Publication number: WO1995003057A1
Application number: PCT/JP1994/001159
Authority: WO
Inventors: Sumio Arai; Cheng-Chuang Tseng
Original assignee: Chugai Seiyaku Kabushiki Kaisha
Priority date: 1993-07-20
Filing date: 1994-07-15
Publication date: 1995-02-02
Also published as: TW264389B; AU7194894A

Abstract

An endotoxin neutralizer containing as the active ingredient a tetracycline antibacterial or a derivative thereof, such as tetracycline, oxytetracycline, demethylchlorotetracycline, doxycycline, minocycline, chlorotetracycline, or a salt thereof; a preventive or remedy for diseases caused by endotoxin, except for the use as an antibacterial; and a preventive or remedy for endotoxin shock.

Description

Specification

Endotoxin neutralizer

[Technical field]

The present invention relates to an endotoxin neutralizing agent containing a tetracycline antibacterial agent or a derivative thereof as an active ingredient. The present invention is used as a preventive / therapeutic agent for various diseases caused by endotoxin and endotoxin shock.

[Background technology]

Endotoxin (bacterial endotoxin) is a component of the outer membrane structure of the cell wall of Gram-negative bacteria. Its major component, lipopolysaccharide (1S), is a complex of polysaccharide and lipid. And its toxicity has been shown to be localized to 1 ipidA.

Endotoxin release in the body causes a variety of symptoms, including fever, chills, horror, and reduced blood pressure, and eventually multiple organ failure (MOF), including diffuse intravascular coagulation (DIC) and renal and hepatic failure It is a big problem that can cause shock and death in patients. It is thought that the pathology of these diseases progresses due to activation of the complement system and coagulation-fibrinolytic system by endotoxin stimulation and interaction with cytokines produced from monocytes, neutrophils, vascular endothelial cells, and the like. In particular, cachectin tumor necrosis factor (T NF) has been shown to cause serious side effects such as lowering of blood pressure in clinical trials, and it has been found that large doses to rats cause shock, metabolic acidosis, and organ damage, resulting in death. It has been attracting attention as a mediator in endotoxin shock. Currently, Gram-negative bacterial infections are the leading cause of morbidity and mortality, especially in hospitalized and immunocompromised patients (immunocompromedostos). Antibiotic treatment is effective in inhibiting bacterial growth through its antibacterial action, but is not effective at neutralizing endotoxin-based biological effects released upon lysis. Thus, despite the significant improvement in the antimicrobial activity of antibiotics, mortality associated with endotoxemia remains high. Thus, there is a need for antimicrobial chemotherapy and infusion therapy, as well as adjuvant therapies to counteract the adverse effects of endotoxin.

To date, polymyxin B has been identified as a substance that blocks the harmful effects of endotoxin. However, its use is limited, for example, topical administration due to its high nephrotoxicity and neurotoxicity. Also, large doses of dalcocorticoid have protective effects in experimental endotoxin shock models. Thus, combination treatment of antibiotics and methylprednisolone was attempted in patients with sepsis and septic shock. However, it has been reported that the usefulness of the combination is not recognized, and that the prevalence of secondary infection is increased by the administration of methylprednisolone. Further, recently, studies on anti-1ipid A antibody ゃ anti-TNF antibody and the like have been attempted, but a reliable adjuvant therapy has not yet been established.

On the other hand, tetracycline antibacterial agents have an extremely wide antibacterial spectrum and have been frequently used for various infectious diseases. Its antibacterial effect is to inhibit bacterial protein synthesis by inhibiting the binding of bacterial aminoacyl-tRNA to liposomes, and its effect on LPS, an outer membrane component of Gram-negative bacteria, is not known. Not. Today, Gram-positive bacteria such as Streptococcus pneumoniae and Streptococcus haemolyticus-Gram-negative bacteria such as Escherichia coli, Klebsiella pneumoniae, and Shigella have become resistant to tetracycline-based antibacterial agents. It has been limited to resistant Staphylococcus aureus.

The present inventors have been studying methods for preventing or treating endotoxin shock and found that the ability of LPS to induce TNF is blocked by existing tetracycline antibacterial agents. The present invention has been completed by further studying this finding.

[Disclosure of the Invention]

The present invention provides: (1) a neutralizing agent for endotoxin containing a tetracycline antibacterial agent or a derivative thereof as an active ingredient; and (2) prevention of a disease caused by endotoxin containing a tetracycline antibacterial agent or a derivative thereof as an active ingredient. And therapeutic agents (except when used as antibacterial agents). And ③ a prophylactic or therapeutic agent for endotoxin shock containing a tetracycline antibacterial agent or a derivative thereof as an active ingredient.

In the present invention, the tetracycline-based antibacterial agent is tetracycline (4-dimethy 1 amino-1,4,4a, 5, oa, 6,11,12a-oc t ahyd r o-3, 6, 10, 12, 12 ap en t ahy d r oxy-6 — me t hy 1— 1, 1 1 di oxo— 2—carboxami de) It refers to a group of antibiotics with four 6-membered rings as the basic skeleton and their derivatives. Specific examples include tetracycline, oxytetracycline, demethylchlorotetracycline, doxycycline, minocycline, quintetratetracycline, and salts or derivatives thereof.

In practicing the present invention, the antibacterial activity of the tetracycline used does not need to be particularly problematic, but only the neutralizing activity against endotoxin. This means that the use of this drug is not restricted even in the current clinical setting where many Gram-negative bacteria have become resistant to tetracycline antibiotics. Further, a tetracycline derivative having no antibacterial activity can be used in the present invention. Such tetracyclines have the advantage that the problem of inducing resistance to bacteria does not occur. The present invention is a method using a tetracycline-based compound as an endotoxin neutralizing agent that inhibits a biological response to endotoxin stimulation. It can be used to ameliorate the pathology of temporary or persistent endotoxemia. Examples of endotoxemia in which the neutralizing agent for endotoxin of the present invention can be used include trauma, burn, peritonitis, biliary tract infection, liver abscess, urinary tract infection, infectious abortion, pneumonia, and sepsis associated with Gram-negative bacterial infection. Can be In addition, endotoxemia due to mechanical manipulations for treatment and examination and endotoxin absorption from intestinal flora due to deterioration of the underlying disease include liver cirrhosis, fulminant hepatitis, surgical operations such as hepatectomy and digestive organs , Suture failure, ulcerative colitis, acute inflammation and the like. Among these diseases, some tetracycline antibiotics have already been indicated for sepsis, trauma, burns, postoperative secondary infections, pneumonia, urinary tract infections, and biliary tract infections. However, these were used according to the so-called antibacterial activity expressed by the minimum inhibitory concentration (MIC) of bacteria, and there is no known use as a neutralizing agent of endotoxin action.

The present invention is further used as an agent for preventing and treating a disease caused by endotoxin. Endotoxin-induced diseases include sporadic intravascular coagulation (DIC), adult respiratory distress syndrome (ARDS), and multiple organ failure (MOF), including renal and hepatic failure And so on.

The present invention is also used as an agent for preventing or treating endotoxin shock. In the present invention, endotoxin shock refers to an irreversible shock state exhibiting acute circulatory failure based on endotoxemia. These are more likely to occur due to severe gram-negative bacterial infections in cancer and immunocompromised patients, as well as contaminated blood transfusions, fluids, and long-term catheter placement.

[Best Mode for Carrying Out the Invention]

In the present invention, the administration of the tetracycline compound is appropriately selected from oral, injection (intramuscular, intravenous, intravenous drip), topical, and the like. The dose is appropriately selected depending on the route of administration, purpose of use (prevention or treatment), and symptoms, but is preferably 100 mg to 1 g (therapeutic agent) and 2 mg to 30 mg (the prophylactic agent). Generally, when used as a prophylactic agent, a relatively low dose is often required.

The tetracycline compound of the present invention can be converted into a pharmaceutically acceptable salt with a suitable acid. Suitable acids include hydrochloric acid, nitric acid, metaphosphoric acid and the like. For oral administration, it can be made into tablets, granules, powders, capsules, suspensions, emulsions, syrups, etc.Addition of excipients, binders, bulking agents, etc. commonly used in formulation Things may be included. For injection, it may be in the form of an aqueous solution, suspension, oily or water-soluble emulsion, but is usually prepared as an aqueous solution such as sterile water or physiological saline. If necessary, commonly used solubilizers, stabilizers, preservatives and the like may be added.

Hereinafter, the present invention will be described specifically with reference to Examples (Experimental Examples), but the present invention is not limited thereto.

【Example】

(Experimental example 1)

Inhibition of TNF-inducing ability of LPS by tetracycline antibacterials (invitro)

Materials and methods: intraperitoneally in I CR mice induced with Chio glycolate macrophage chromatography di (10 ⁵ Zm 1) the addition of E. coli LP S (0111 B 4) 1 μ g / 1 and tetracycline antibacterial agent 10 gZm 1 And incubate at 37 ° C for 4 hours After culturing in a culture vessel, TNF activity in the culture supernatant was measured as cytotoxic activity against L cells.

Results: As shown in Table 1, minocycline, doxycycline, chlortetracycline, oxytetracycline, and tetracycline all significantly inhibited TNF production.

Tetracycline TNF activity (UZm 1)

Antibacterial agent (% rejection)

Minocycline 200 (87.5)

Doxycycline 220 (86.3)

Chlortetracycline 380 (72.4)

Oxytetracycline 450 (71.75)

Tetracycline 240 (85.0) Control 1600 (0)

(Experimental example 2)

Inhibition of TNF-inducing ability of LPS by tetracycline antibacterial agents (invivo)

Materials and Methods: ICR mice (5-week-old 25-28 g) were intraperitoneally injected with 100 r of LPS and 50 Or or 1000 r of tetracycline (TC), and TNF in the blood of the mice 4 hours later The amount was measured. After a mixture of LPS and TC was cultured at 4 ° C for 4 hours, the mixture was intraperitoneally administered. TNF was measured by cytotoxicity to L cells.

Results: As shown in Table 2, administration and addition of tetracycline clearly suppressed TNF production as compared to the control. Table 2

TNF activity (UZm 1) control 58

TC 500 r, LPS 00 r (simultaneous strike) 06

TC 1000 r, LPS 100 r (simultaneous shot) 37

TC 500 r, LPS 00 r

(Pre-incubation)

TC 250 r, LPS 100 r 83

n = 6

(Experimental example 3)

Effect of tetracycline on LPS-induced mouse mortality

(Iv) As a result of examining the life-span effect of tetracycline in the system in which LPS and 1000 r were administered to the ICR mouse of Experimental Example 2, all the mice died within 3 days in the tetracycline-untreated mice. Administration of tetracycline 160; zg did not cause any mouse death.

Claims

The scope of the claims

1. An endotoxin neutralizing agent containing a tetracycline antibacterial agent or a derivative thereof as an active ingredient.

2. A preventive and therapeutic agent for diseases caused by endotoxin containing a tetracycline antibacterial agent or a derivative thereof as an active ingredient (except when used as an antibacterial agent).

3. An agent for preventing or treating endotoxin shock, comprising a tetracycline antibacterial agent or a derivative thereof as an active ingredient.

4. The method according to any one of claims 1, 2 and 3, wherein the tetracycline antibacterial agent or a derivative thereof is tetracycline, oxytetracycline, demethylchlorotetracycline, doxycycline, minocycline, chlortetracycline, a salt thereof, or a derivative thereof. Or the drug according to item 1.