JPH0489428A - Polymorphonuclear leukocyte activator - Google Patents

Abstract

PURPOSE:To obtain the title activator capable of treating hardly curable infectious diseases such as periodontal diseases, athlete's foot, and opportunistic infection, comprising a histamine H1 acceptor antagonist and/or a histamine H2 acceptor antagonist as active ingredients. CONSTITUTION:A polymorphonuclear leukocyte activator comprising histamine H1 acceptor antagonist such as diphenhydramine, pyrilamine, antazoline, promethazine or chlorpheniramine and/or histamine H2 acceptor antagonist such as brimamid, metiamide, cimetidine or ranitidine as active ingredients. The histamine acceptor antagonist recovers phagocytic activity of polymorphonuclear leukocyte weakened by histamine and improves gingivitis, an example of inflammation occurring by usual microorganisms. The activator is especially effective for opportunistic infection against which antibiotics and germicides are not effective.

Description

[Detailed Description of the Invention] [Field of Industrial Application] This invention relates to polymorphonuclear leukocyte activators, and more specifically, the present invention relates to polymorphonuclear leukocyte activators that activate polymorphonuclear leukocytes, which are one of the body's defense functions against infection by bacteria, etc. This invention relates to drugs for preventing and treating various infectious diseases.

[Conventional technology]

Conventionally, various antibiotics have been used to treat infectious diseases caused by bacteria and the like. For example, periodontal disease is thought to be caused by bacteria, and antibiotics such as tetracycline and bactericidal agents such as metronidazole are used as therapeutic agents.

On the other hand, phagocytosis of bacteria by polymorphonuclear leukocytes is known as one of the defense reactions of living organisms against such infections.

[Problem to be solved by the invention]

However, antibiotic treatment for infections may not be sufficiently effective. For example, periodontal disease is an infection caused by bacteria that normally reside in the oral cavity, so when treated with antibiotics or bactericides, the number of bacteria decreases during the period of drug administration, and the symptoms improve; There was a problem that if the treatment was discontinued, the resident bacteria would re-proliferate and the infection would recur.

Furthermore, if antibiotics, disinfectants, etc. are used continuously over a long period of time to deal with this problem, there is a high risk that drug-resistant bacteria will appear or bacterial replacement diseases such as candidiasis will occur.

[Means to solve the problem]

Therefore, the present inventors focused on the antiphagic bactericidal action of polymorphonuclear leukocytes, which play an important role in suppressing the occurrence of periodontal disease in the body's natural immunity, and aimed to activate this or We have been conducting extensive research on the idea that by normalizing the decrease, we can promote the sterilization and elimination of pathogenic bacteria at the infection focus and prevent or treat various infectious diseases. Focusing on histamine, which is widely known as a type of inflammatory mediator, after repeated studies it was revealed that the phagocytic activity of polymorphonuclear leukocytes was suppressed by histamine, and was further attenuated by histamine receptor antagonists. The inventors have discovered that the phagocytosis of polymorphonuclear leukocytes can be restored and that infectious diseases such as periodontal diseases can be treated, and the present invention has been completed.

That is, the present invention provides a polymorphonuclear leukocyte activator containing histamine H, a receptor antagonist, and/or a histamine H2 receptor antagonist as an active ingredient.

In the present invention, the histamine H1 receptor antagonists include:
Examples include, but are not limited to, diphenhydramine and doxylamine of the aminoalkyl ether type, birilamine of the ethylenediamine type, antazoline of the imidacilline type, promethacin and alimemazine of the henotiazine type, homochlorcifrizine of the homopiperazine type, chlorpheniramine of the propylamine type, and pyridoidene. Phenyndamine type, cycloheptane type cycloheptane type. The histamine H3 receptor antagonists are also not limited, and include brimamide, methyamide, cimetidine, ranitidine, and the like. In addition, these histamine receptor antagonists are
One kind or a mixture of two or more kinds can be used.

The results of tests on the polymorphonuclear leukocyte activation effect and infectious disease treatment effect using representative histamine receptor antagonists, which are the active ingredients of the present invention, are shown.

(a) Attenuation of the phagocytosis of polymorphonuclear leukocytes by histamine and the effect of histamine receptor antagonists on it Blood was collected from the heart of a male Hartley guinea pig weighing approximately 500 g, and the leukocyte fraction was separated using Monopoly Separation Solution (FLOW). HBSS() tanks' bala was collected and polymorphonuclear leukocytes were further separated by Percoll density centrifugation.
After washing with nced 5alt 5 solution), it was confirmed by a trypan blue exclusion test that 95% or more of the cells were living cells, and the cells were dispersed in HBSS and used as a polymorphonuclear leukocyte sample.

Separately 0.5% FITC in carbonate buffer (p) 19.1)
(f 1uoresceinisothiocyana
Zymodene (ly) labeled by reacting with te) for 2 hours
2.5X of dispersing mosan') into IBSS
Guinea pig serum 11IIAI was added to each 10" plate and incubated at 37°C for 30 minutes to perform obsonization with complement.

The above polymorphonuclear leukocytes (final concentration: 1.6XIO' cells/d
After preincubating the drug and the drug in a test tube at 37°C for 20 minutes, the same Zymosan mentioned above (final concentration: 2.4 x 10' cells/tube) was added and incubated at 37°C and 0°C for 20 minutes. I fed him. Then, the average fluorescence intensity per 1000 cells was determined using a flow cytometer (BB (manufactured by IN5ON: TONOIC)) and
The difference between the temperature and 0°C was defined as the hypophagic activity.

The drugs used were histamine hydrochloride, pyrilamine hydrochloride, a typical H3 receptor antagonist, and cimetidine hydrochloride, a typical H3 receptor antagonist.

The drug was evaluated as follows. First, it was confirmed that the phagocytic activity of polymorphonuclear leukocytes not treated with drugs was suppressed by histamine treatment (1 mM).

Next, at the same time as histamine, various antagonists (10,100°1
The hypophagic activity upon treatment with 000 μM) was measured, and the presence or absence of recovery was evaluated. The results are shown in Figure IA and Figure IB. H
Both pyrilamine hydrochloride, an H1 receptor antagonist, and cimetidine hydrochloride, an H2 receptor antagonist, restored oligophagic activity in a dose-dependent manner, and at 100 μg/ml the activity became comparable to the control.

(Company) The effect of histamine receptor antagonists on spontaneous gingivitis in dogs.Female peagle dogs weighing approximately 4 kg were fed dog feed softened with water for one year, and oral resident bacteria were found to be the cause. Naturally caused gingivitis.

Under thiobental anesthesia, once a day, use a root canal syringe to inject 1ml of the drug into the gingival sulcus of this inflamed area! was injected. The drugs used were chlorpheniramine maleate, an H11 receptor antagonist, and physiological saline solutions (containing 10% ethanol) of cimetidine hydrochloride and ranitidine hydrochloride, both receptor antagonists, at a concentration of 0.1%.

The dog's jaw was divided into four parts, horizontally and vertically, and a control physiological saline solution and three types of drugs were applied to each quarter. The treatment was carried out for 5 consecutive days, and the degree of inflammation was evaluated under halothane inhalation anesthesia on the first day of the experiment, and on the 3rd and 4th days.

The degree of inflammation was evaluated by measuring the amount of gingival crevicular exudate (periodron value); the amount of gingival crevicular exudate increases as gingival inflammation worsens, and is effective in quantifying the degree of inflammation. . The results are shown in FIGS. 2A to 20. Cimetidine hydrochloride, an H22 receptor antagonist, significantly suppressed inflammation, and ranitidine hydrochloride showed a tendency to suppress inflammation.

(C) Effect of histamine receptor antagonist on severe gingivitis in dogs experimentally induced with ligature Significant 4
A female peagle dog (kg) was brushed once a day for one month to ensure healthy gingiva, and then a surgical suture was wrapped as a ligature around the neck of the test gingival tooth under pendoparbital anesthesia. Severe gingivitis developed one week later.

This was treated with drugs in the same manner as described above. The drugs used were mepyramine hydrochloride, an HI receptor antagonist, cimetidine hydrochloride, an H22 receptor antagonist, and indomethacin, a nonsteroidal anti-inflammatory drug, each at 0.8% (2
0ωM), 0.5% (2001M), 0.07% (
2mM) saline solution (containing 10% ethanol)
1ml per gingival sulcus once a day! Injected. Measurements were performed in the same manner as before the induction of gingivitis, on the day of starting drug treatment, and on the 2nd, 4th, and 6th days.

The results are shown in Figures 3A to 3D. The H receptor antagonist pyrilamine hydrochloride was administered on the 4th day, the H22 receptor antagonist cimetidine hydrochloride was administered on the 6th day, and the non-steroidal anti-inflammatory drug indomethacin was administered as a control. significantly suppressed inflammation on the 4th and 6th day. No improvement in inflammation was observed in areas that were not treated with drugs.

As shown in the experimental examples (a) to (C) above, histamine receptor antagonists, H receptor antagonists, and H22 receptor antagonists all restored the phagocytic activity of polymorphonuclear leukocytes that had been attenuated by histamine. . It also improved gingivitis, which is an example of inflammation caused by resident bacteria. Therefore, histamine receptor antagonists are useful as polymorphonuclear leukocyte activators. In addition, the polymorphonuclear leukocyte activator of the present invention can be used to treat many infections accompanied by inflammation in addition to periodontal diseases, especially opportunistic infections caused by indigenous bacteria for which antibiotics and bactericidal agents are not effective. etc. are more suitable.

The polymorphonuclear leukocyte activating agent of the present invention contains suitable excipients, carriers,
Using a diluent, etc., tablets, capsules, granules, powders, liquids that can be injected into the gingival sulcus, ointments, dissolvable sticks impregnated with drugs, plasters, poultices, injections, suppositories, etc. The dosage form can be administered orally or parenterally. These formulations can be made by methods known per se. For example, a preparation for oral administration may include a histamine receptor antagonist in an excipient such as starch, mannitol, or lactose;
Binders such as sodium carboxymethyl cellulose and hydroxypropyl cellulose; Disintegrants such as crystalline cellulose and calcium carboxymethyl cellulose; Talc,
It can be manufactured by appropriately combining and formulating a lubricant such as magnesium stearate; a fluidity improver such as light anhydrous silicic acid, and the like. In addition, in preparing external preparations such as ointments and poultices, known bases such as vaselines, lanolin, paraffin, silicones, animal and vegetable oils, etc. can be used.

The amount of H, receptor antagonist and/or H22 receptor antagonist added to the polymorphonuclear leukocyte activating agent of the present invention may be any amount as long as these are in amounts effective for activating polymorphonuclear leukocytes. ,
Usually, for internal use, the amount is preferably 1 to 1000 cm/day, and for external use, it is preferably 0.01 to 60% by weight. Furthermore, in addition to these H11 receptor antagonists and 82 receptor antagonists, other drugs that can be combined can also be formulated at the same time. In particular, the combination of the polymorphonuclear leukocyte activating agent of the present invention and antibiotics is effective.

〔Example〕

Next, the present invention will be described in detail with reference to Examples, but the present invention is not limited thereto.

Example 1 Periodontal disease therapeutic agent Mepyramine hydrochloride 0.1 Cimetidine hydrochloride 0.1 Total 100 (wt%) The above liquid preparation was injected into the gingival sulcus using a root canal syringe.

Example 2 Periodontal disease therapeutic agent Mepyramine hydrochloride 0.1 Ranitidine hydrochloride 0.1 Hydroxyethyl cellulose
1.0 Total 100 (% by weight) The above viscous liquid is injected into the gingival sulcus using a root canal syringe.

Example 3 Periodontal disease therapeutic agent Ranitidine hydrochloride 1.0 Tocopherol acetate 5. ON lauroyl arginine
0.5 ethyl ester total 100 (wt%) The above composition was added to 10 times the amount of water to make an emulsion,
Work hard.

Example 4 Hemorrhoid disease treatment agent cimetidine hydrochloride 0.2 quercetin
0.1 Chlorhexidine hydrochloride
0.05 total 100 (wt%) The above composition is heated and melted and mixed to form a bullet shape to make a crude drug.

Example 5 Skin disease therapeutic agent Mepyramine hydrochloride 0.1 Ranitidine hydrochloride 0.1 Glycyrrhetinic acid
0.02 tocopherol acetate 0.1 total
100 (wt%) Apply the above ointment to the affected skin area.

〔Effect of the invention〕

According to the present invention, various diseases in which the activity of polymorphonuclear leukocytes is decreased due to bacterial infection, such as periodontal disease, intractable infections such as athlete's foot, and hemorrhoids, and opportunistic infections can be effectively treated. be able to.

[Brief explanation of the drawing]

Figure IA and Figure IB are diagrams showing the effects of cimetidine hydrochloride and birilamine hydrochloride, respectively, on the decrease in polymorphonuclear leukocyte anemia (mean fluorescence intensity) caused by histamine. FIGS. 2A, 2B, and 20 are drawings showing the effects of physiological saline, ranitidine, and cimetidine hydrochloride on the amount of gingival crevicular fluid (periodrone value) in spontaneous gingivitis. Figures 3A to 3D show the amount of gingival crevicular fluid (periodron value) in dog gingivitis caused by ligature.
2 is a diagram showing the effects of physiological saline (control), cimetidine hydrochloride, pyrilamine hydrochloride, and indomethacin on the sterilization.

Claims (1)

[Scope of Claims] 1. A polymorphonuclear leukocyte activator containing a histamine H_1 receptor antagonist and/or a histamine H_2 receptor antagonist as an active ingredient. 2. A periodontal disease preventive/therapeutic agent containing the polymorphonuclear leukocyte activator according to claim 1.