JPH02250827A - Antibacterial agent, anti-inflammatory drug and external medicine for cutis - Google Patents

Abstract

PURPOSE:To obtain antibacterial agent, anti-inflammatory drug and external medicine for cutis containing 6-hydroxyluteolin having strong antibacterial activity, wide antibacterial spectrum, high safety and strong anti-inflammatory action. CONSTITUTION:The aimed antibacierial agent, anti-inflammatory drug and external medicine for cutis contain 6-hydroxyluteolin expressed by the formula as active ingredient. 6-hydroxyluteolin, as naturally derived, e.g. is obtained by extraction of leaf of Catalpa bignonioides in a form of glucoside and hydrolyzing with acid, or able to be obtained by synthesis of 3,6-dihydroxy-2,4- dimethoxyacetophenone, etc., as a raw material. Said 6-hydroxyluteolin exhibits remarkable histamine release-inhibiting action. Besides, said external medicine for cutis exhibits not only anti-inflammatory effect but also inhibition of pollution or microbism by microorganism in a case of coating on affected part of inflammation.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to an antibacterial valve L anti-inflammatory agent and a skin external preparation.

(Prior Art) Antibacterial agents generally include sulfa drugs such as sulfamine J, sulfadiazine and sulfisomidine, and antibiotics such as penicillin, streptomycin, and tetracycline. Other sterilizing valves L Phenol derivatives and alcohols are used as disinfectants and preservatives. As anti-inflammatory agents, steroidal anti-inflammatory agents such as hydrocortisone, nonsteroidal anti-inflammatory agents such as salicylic acid derivatives, pyrazolone derivatives, and antihistamines such as diphenhydramine are used.

(Problem to be solved by the invention) However, many antibiotics and sulfa drugs have a wide range of side effects, and in particular, as typified by penicillin, their topical use can cause percutaneous sensitization, which can cause unexpected and serious damage. There is a risk of this happening. Furthermore, phenol derivatives and alcohols often do not have sufficient antibacterial activity. As for anti-inflammatory drugs, steroid-based anti-inflammatory drugs have a wide range of side effects such as infection calming, gastrointestinal disorders, and skin hypersensitivity, while non-steroidal anti-inflammatory drugs do not have as strong side effects as steroid-based drugs, but are sufficiently effective. The current situation is that it is not effective.

In view of this situation, the present inventors conducted intensive research and discovered that 6-hydroxylthioline has strong antibacterial activity, a broad antibacterial spectrum, and is highly safe. Further, through investigation of its physiological activity, it was discovered that jujube also has a strong anti-inflammatory effect, leading to the completion of the present invention.

(Means for Solving the Problems) The 6-hydroxylthioline referred to in the present invention may be a synthetic product or a natural product.

There are the following reports regarding the origin of natural products. J.

B. Harborne et al.
Hnonoids Walt.

(American Catalpa), Catalpa bung
ei Mey, Catalpa 5peciosa
Ward, Tecoma australi
It was found in the form of glucoside from the leaves of sR, Br, and 6-hydroxylthioline was obtained by acid hydrolysis (Ph
ytochemiatry, F! , 1643-16
51.1967).

Synthetic products can be obtained as follows, e.g.
．． 3 obtained by partial benzylation of 6-cyhydroxy-2,4-dimethoxyacetophenone (0.1 mol)
-Benzyloxy-6-hydroxy-2,4-dimethoxyacetophenone was reacted with 3,4-bis(benzyloxy)benzoyl chloride (0.12-0.14 mol) in the presence of pyridine at 100-120°C. Thereafter, the reaction solution was poured into ice-hydrochloric acid, and the precipitated oil was extracted with ethyl acetate. The extract was dissolved in sodium carbonate using the aqueous method.
After washing with water, the solvent is distilled off to obtain a crude ester. After sufficiently drying the obtained crude ester, it is dissolved in pyridine, pulverized potassium hydroxide (0.5 to 1 mol) is added, and the mixture is reacted with stirring at 55 to 60°C for 4 to 5 hours. Ice is added to this to dissolve excess potassium hydroxide, and the mixture is poured into ice-hydrochloric acid. Add ethyl acetate to the mixture and stir to dissolve the oil. The precipitated yellow crystals were collected by filtration and dried with a water pipe to give a yellow crystal diketone derivative (mp135-1).
36 and 143-144°C). Total yield 60-7
0%.

Dissolve this diketone in acetic acid under heating, add sodium acetate (approximately 10% of acetic acid), and then
Sufficient water is added to the reaction mixture heated for 0 hours, ether is added to form two layers, the oil is dissolved, and the mixture is left in an ice chamber to precipitate colorless needle crystals. Filter this out.
Dry, 3',4',6-tris(benzyloxy)
-5,7-simethoxyflavone (+ap 89-91
°C) yield 90-95%.

10% of this flavone in ethyl acetate-methanol mixture
Hydrogenolyzed using palladium on carbon to obtain 3°, 4', 6-
Drihydroxy-5,7-simethoxyflavone (mp2
98-300°C) was obtained quantitatively. When this is further acetylated with acetic anhydride, triacetate (mp240~
241℃), 30-40% of this triacetate
(W/V) Dissolve in anhydrous aluminum chloride-acetonitrile solution (more than 6 molar ratio) and heat to 70 °C for 36-48 hours.The reaction mixture is poured into ice-hydrochloric acid and heated for 2-3 hours.Hydrolysis do. The precipitated yellow precipitate was collected by filtration, washed with water, and the desired 6-hydroxylthioline (mp) was obtained at 300°C.
) was obtained. Yield 85-95%.

The pure product is converted into pentaacetate by acetic anhydride-pyridine (
mp 246-247°C), purified by recrystallization from a chloroform-methanol mixture, and then hydrolyzed with methanolic hydrochloric acid. IHNMR (δ value;
ppm): (DMSO-ds) 6.63 (IH,
8, C3-H), 6.54 (IH, a, C@-H),
7.39(1)1. d, J=2.5H2, C2゜-〇), 6.89(1)1. d, J=8.511z, C
s・-H), 7.41 (111, dd, J=8.5,
2.5+1z, C6・-H), 12.81(IH, a
, CB-OH), uv': λssx(logl
); (EtOH)281(4,22), 353nm
(4,33); (EtOH-AICll)257
ah (4,11), 294 (4,22), 378
(4,42);(1,tOH-NaOAc)375(4
, 30).

The 6-hydroxylthioline of the present invention does not necessarily have to be an isolated or synthesized pure product, and mixtures containing it, partially purified extracts, etc. can also be used.

The antibacterial formulation of the present invention, characterized by containing 6-hydroxylthioline as an active ingredient, can be used as an anti-inflammatory agent or skin preparation for external use, as long as it does not impair its effectiveness. k Bond Qi L Decay Qi L Namezawa Qi L
Coloring Qi L Flavoring Correcting Qi 1. It can be formulated by a conventional method by adjusting the pH, solubilizing L, suspending L, buffering L, stabilizing, and adding preservatives.

The antibacterial agent and anti-inflammatory agent of the present invention are: Extract L, Turning powder into granules, Powder, eye ointment L, Suspension/milk, Plaster L, Zombie, Fine granule L, Tablet, Syrup, Dip L, Pre-press L. Injection number 1 Eye drop number L Troche number L Ointment number L Pap number number 1 Liniment number number 1 It can be used as a lotion, etc. In addition, the skin external preparation of the present invention is a plaster Qi L.
Ointment Qi L Seating valve L Alcohol Qi L Tincture Qi Pap Qi L
Liniment Qi L can be used as a lotion, etc. Note that the products to which 6-hydroxylthioline of the present invention is added are not limited to the above-mentioned products.

The antibacterial agent and anti-inflammatory agent of the present invention are administered orally or parenterally (intramuscularly, subcutaneously, intravenously, as a waste drug, etc.). Moreover, the skin external preparation of the present invention is applied to the skin in an effective amount several times a day.

The stake of the invention! 14k The dosage of anti-inflammatory agents and topical skin preparations varies depending on the symptoms of the disease, age, etc., but is usually
Approximately 30 to 6 as 6-hydroxylthioline per day,
000 rag, preferably 100 to 3.000 rag.

Next, the present invention will be explained in more detail with reference to Examples. Note that the present invention is not limited to these.

(Example) Cold” 1 case” 23% combination ointment ■ Purified lanolin 5.0 parts ■ White beeswax 5.0 ■ 6-Hydroxylthioline 3.0 ■ White petrolatum
87.0 [Manufacturing method] After heating and melting each component and mixing, the product is cooled.

Emgo 1 Tablet■6-Hydroxylthioline 20.0 parts■Starch 1000■Refined white sugar
1O90■Carboxymethylcellulose calcium 1O00■Microcrystalline cellulose 35.0■Polyvinylpyrrolidone 5.0■Talc
10.0 [Production method] Ingredients ■ to ■ were mixed, and then the aqueous solution of ■ was added as a binder and granulated by the usual method. After adding ■ as a lubricant to this, the mixture was compressed into tablets of 100 mH each. Tablets 9 and 51 Powder ■ 6-Hydroxylthioline 50.0 parts ■ Microcrystalline cellulose 25.0 ■ Starch
25.0 [Manufacturing method] Mix ingredients ■ to ■ and make powder by conventional method. Nimu ■ 1 Capsule ■ 6-hydroxylthioline 40.0 parts ■ Lactose 40.0 ■ Starch
1O10■ Talc
5.0■Magnesium stearate
5.0 [Manufacturing method] Ingredients (1) to (2) are passed through a sieve, mixed, and put into a hard capsule to form a product.

(Effect of the invention) Next, how excellent is the antibacterial activity of the antibacterial agent of the present invention?
A comparative test was conducted between the antibacterial agent of the present invention and methyl paraoxybenzoate, a conventional antibacterial agent.
The results are also shown in Table 1.

1111 Agar plate dilution method (Chemotherapy, 29.7
6-79.1981), the minimum inhibitory concentration (MIC) for the following test bacteria was measured under the following culture conditions.
The sample was dissolved in dimethyl sulfoxide and then added to the culture medium to a predetermined concentration.[Test bacteria] Staphylococcus aureus
FDA 209P) 2 Propionibacterium acnes
)3 Psedomonas aeruginosa
ATCC9G27) 4 Staphylococcucus epide
rmidis) 5 Trichophyton rubrum 6 Streptococcus thermophilus
ccus thermophilus)Culture conditions
(ar GAM a(ar Table 1 Minimum inhibitory concentration (MIC) 0.0+ 0.02 0, Ol O,0025 0.01 O2O3 measurement result 0.2 0.2 0.2 0.2 0.1 0. 1 As is clear from Table 1, the 6-hydroxylthioline of the present invention exhibited excellent antibacterial activity.

Next, Experimental Examples 2 and 3 show the results of a comparative test between the anti-inflammatory agent of the present invention and indomethacin, a conventional anti-inflammatory agent, to show how excellent the anti-inflammatory effect of the anti-inflammatory agent of the present invention is.

The anti-inflammatory effect of carrageenan was investigated using the rat carrageenan paw edema method. That is, using male rats weighing around 150 g,
6-hydroxylthioline (50 B/kg body weight) or indomethacin (10 mg/kg body weight) was administered intraperitoneally, and 0.5 hours later, 0.1 ml of 1.0% carrageenan was administered as an inflammatory agent to the sole of the right foot. Carrageenin administration 1.
The constant volume of the right hind paw was measured after 2, 3, 4, 5.6 and 24 hours. Physiological saline was used as a control. The measurement results are shown in FIG. 1. As is clear from FIG. 1, 6-hydroxylthioline showed a strong anti-inflammatory effect.

Report by Yuhirai et al. (Pharmaceutical Journal, 37.374-380.
(1983), the inhibitory effect on histamine release on mast cells collected from the intraperitoneal cavity of rats was measured.

That is, Table 2 shows the experimental results in which the effect of suppressing histamine release by 4 g/80 of the 4pp+* compound was determined as the release inhibition rate.

Table 2 Histamine release inhibition effect Sample concentration (%) Histamine release inhibition rate (%) 6-
Hydroxylthioline 0.01 10
00.003 1
000.001
54 Indomethacin 0.1 83
0.01 36
From the results shown in Table 1, it was confirmed that 6-hydroxylthioline exhibited a remarkable effect of inhibiting histamine release, and was more effective than indomethacin, an existing anti-inflammatory agent.

Since the skin external preparation of the present invention has excellent antibacterial and anti-inflammatory effects, when applied to an inflamed area, it not only exhibits an anti-inflammatory effect but also can prevent infection by microorganisms in the affected area.

Next, the safety of 6-hydroxylthioline of the present invention will be described.

In oral toxicity tests using mice (ICR system), the L
The D50 value is more than 3,000 mg8, and it is recognized that it is a highly safe substance with no acute toxicity.
A time occlusion patch test was conducted, and the average value of the results 1 hour, 24 hours, and 48 hours after removal of the patch test adhesive was used for evaluation. No formation of erythema, dermatitis, or edema was observed, confirming that 6-hydroxylthioline is a substance with low skin irritation. The sample was tested using a 50% aqueous dispersion.

Regarding contact sensitization, Maxi-m using guinea pigs
Freund
The cells were sensitized by subcutaneous administration of a sample containing 1% concentration of 6-hydroxylthioline in a base emulsified with ''s Complete Adjuvant and physiological saline, and by transdermal administration of a sample containing 1% concentration of 6-hydroxylthioline in a white petrolatum base. , Chal at 0.2 and 2.0% concentrations using white petrolatum base.
leBet Results All 12 guinea pigs used in the test were negative, confirming that the contact sensitization of 6-hydroxylthioline was extremely low.

[Brief explanation of drawings]

Figure 1 shows 6-
These are the results of an anti-inflammatory test on hydroxylthioline. The vertical axis shows the edema paper, and the horizontal axis shows the time after carrageenan administration. Patent applicant Nonoyo Shoji Co., Ltd. Time after administration of carrageenan (hr)

Claims (3)

[Claims] (1) An antibacterial agent characterized by containing 6-hydroxylthioline represented by the general formula [I] as an active ingredient. (2) An anti-inflammatory agent characterized by containing 6-hydroxylthioline represented by the general formula [I] as an active ingredient. (3) An external skin preparation characterized by containing 6-hydroxylthioline represented by the general formula [I] as an active ingredient. [I] ▲ Contains mathematical formulas, chemical formulas, tables, etc. ▼