JPH09188621A - Anti-mrsa active medicinal composition containing xanthatin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an anti-MRSA(methicillin-resistant Staphylococcus aureus) active medicinal composition containing a component contained in 'cangerzi' as a Chinese medicine hardly having adverse effect as an active component. SOLUTION: This anti-MRSA medicinal composition contains xanthatin expressed by the formula and contained in 'cangerzi' as a fruit of Xanthium strumarium as an active component. Xanthatin is obtained by, e.g. refluxing crushed leaves of Xanthium strumarium in 50% ethanol, suction-filtrating in a warm state, concentrating the filtrate under reduced pressure, adding ether to the resultant ethanol extract and subjecting to liquid-liquid partition extraction, concentrating and drying up ether layer at 70 deg.C under reduced pressure, fractionating the ether extract by a column charged with silica get in using n-hexane-ethyl acetate and gathering a fraction having an antibacterial activity. Xanthatin can be prepared as an oral medicine or an external medicine having anti-MRSA bacterial action. The external medicine can be used by adding to a commercial disinfectant solution. About 0.1-10% of xanthatin can be contained in the composition. The oral medicine is preferably administrated at about 10-100 mg per one time to an adult (about 70kg).

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an anti-MRSA active pharmaceutical composition containing xanthatin as an active ingredient.

[0002]

BACKGROUND OF THE INVENTION Penicillin, the first antibiotic, has a β-lactam ring and exhibits excellent efficacy against Staphylococcus. However, penicillinase (β-lactamase) is relatively easy for staphylococci
Productivity is induced and the β-lactam ring is cleaved and inactivated. It is called a penicillin resistant bacterium. Regarding this penicillin-resistant bacterium, for example, it seemed that clinically most of it was solved by research and development of penicillinase-resistant penicillin such as methicillin and cephem antibiotics, but all β-lactam agents were ineffective in methicillin-resistant yellow grapes. Cocci (Methicillin-resistant
Staphylococcus aureus : MRSA) has appeared. That is, MRSA is a multidrug-resistant Staphylococcus aureus that is widely resistant to not only penicillin-based antibiotics but also cephem-based antibiotics and aminoglycoside antibiotics. In recent years, as a result of the abuse of third-generation cephem antibiotics, which have weak antibacterial activity against staphylococci, this resistant bacterium has selectively propagated and spread to the hospital, and it has become a major hospital-acquired bacterium. It is becoming a serious social problem. MRSA currently used
There are vancomycin (VCM) and the like as antibacterial agents against infectious diseases, but the short-term bactericidal action of VCM is not strong at all and there is a problem of serious side effects such as audiotoxicity and nephrotoxicity.

The inventors of the present invention searched for compounds having anti-MRSA activity in the ingredients of Chinese herbs that had no or no side effects, and found that Xanthatin, which is an ingredient contained in blue ear sardines. Have anti-MRSA activity. The present invention has been completed based on such findings.

[0004]

The present invention provides an anti-MRSA active pharmaceutical composition containing xanthatin as an active ingredient.

Xanthatin is a species of Xanthium strum.
arium ) is an ingredient contained in the fruit (blue ear). The main components of blue ear ears are fatty oil, xanthinine, xanthine and the like. Onami fir is a plant of the Asteraceae family that grows in the wastelands of Hokkaido, Honshu, Shikoku, Kyushu, and Okinawa, and is distributed in Taiwan, Eurasia, North America, and so on. The fruit is a spindle-shaped spine with a length of about 1 cm, which is often attached to clothes and animals. It is dull and grayish when ripe with no luster, and more recently, naturalized plants that are native to North America are more common. The fruit of Ona fir is called "cangerzi" in Chinese medicine, and it is taken for colds, headaches, etc., for fever and for the purpose of sweating. In addition to the above, in the private sector, raw squeezed juice of foliage is used for insect bites, eczema, scabs, cuts, etc. Also used for allergic rhinitis in China.
Contraindicated in pregnant women, concomitant with Sourikosan, Fukukiko. However, it has not yet been reported that somatoum has anti-MRSA activity.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION

Purification of xanthatin 3.6 kg of crushed blue-eared flea leaves were refluxed with 50% ethanol (36 L) for 2 hours, suction-filtered while warm, and the filtrate was concentrated under reduced pressure. This operation was repeated 3 times, and ether (1.8 L) was added to the concentrated ethanol extract to carry out liquid-liquid partition extraction. The ether layer was concentrated to dryness under reduced pressure at 70 ° C. to obtain a green clay-like substance (24.09 g). The ether extract was layered on a glass column (70 × 200 mm) packed with silica gel (Wakogel C-300) and fractionated with n-hexane-ethyl acetate to obtain a fraction having antibacterial activity. Further, this fraction (15.74 g) was layered on a glass column (25 × 300 mm) packed with silica gel (Kieselgel 60), and fractionated with n-hexane-ethyl acetate (1: 1). O active fraction
Glass column (40 × 75m) packed with DS-Q3 (wako)
m) and fractionated with 50% methanol, then L
Purified with n-hexane-ethyl acetate (1: 1) using iChrosorb Si60 (25 x 250 mm), xanthatin 2
Isolated with a yield of 00 mg.

Xanthatin

Embedded image Colorless needle crystals mp.103-105 ° C IR (KBr): 1762 cm ^-1 (lactone), 1686 cm
^-1 _{(ketone) EI-MS: C 15 H} 18 O 3 (246.1266) [α] D = -8.0 ° (1.0, CHCl 3) 1 H-NMR (400MHz, CDCl 3) δ: 7.06 (1H, d, H-2), 6.19 (1H, d, H-3),
6.28 (1H, dd, H-5), 2.77 (1H, ddd, H-6)
α), 2.23 (1H, ddd, H-6β), 2.56 (1H, m, H-
7), 4.30 (1H, ddd, H-8), 1.82 (1H, ddd, H-
9α), 2.37 (1H, ddd, H-9β), 3.09 (1H, m, H
-10), 6.21 (1H, d, H-13), 5.49 (1H, d, H
-13 '), 1.16 (3H, d, H-14), 2.31 (3H, s,
H-15) J (Hz): 2,3 = 16.1; 5,6α = 9.3; 5,6β
= 3.4; 6α, 6β = 16.6; 6α, 7 = 2.93; 6
β, 7 = 11.7; 7,8 = 9.8; 7,13 = 7.13 '=
3.4; 8.9α = 12.2; 8.9β = 2.4; 9α, 9β
= 12.2; 9α, 10 = 3.9; 9β, 10 = 3.4; 1
0,14 = 7.3 ¹³ C-NMR (100 MHz, CDCl ₃ ) δ: 18.9 (C-14), 27.2 (C-6), 27.9 (C-
15), 29.1 (C-10), 36.6 (C-9), 47.4.
(C-7), 81.5 (C-8), 119.0 (C-13), 12
4.7 (C-5), 138.1 (C-3), 139.2 (C-1),
144.8 (C-2) 148.5 (C-2), 169.7 (C-
4), 198.5 (C-12)

Antibacterial activity against MRSA MRSA (methicillin-resistant Staphylococcus aureus) as a test bacterium
20 strains, MSSA (methicillin-sensitive Staphylococcus aureus) 7
Strains, 3 other Gram-positive bacteria, and 11 Gram-negative bacteria were used. MRSA used as an index for isolation and extraction of active ingredients
The antibacterial activity was measured by the disk diffusion method. About 10 ⁵ CFU (colony forming unit) of MRSA bacterial solution was smeared on an agar plate, and a disc (diameter 13 mm) imbibed with 100 μl of a test sample at various temperatures was placed and incubated at 37 ° C. for 20 hours. After that, it was confirmed whether or not a stop band was formed. The antibacterial activity of the purified preparation was determined by MIC according to the agar plate dilution method defined by the Japanese Society of Chemotherapy. The minimum concentration at which the growth was completely inhibited was defined as the MIC value. The results are shown in Tables 1 and 2.

[0009]

[Table 1] Antibacterial activity of Pseudomonas Et ₂ O extract and xanthatin (XS-1) against methicillin-resistant (MRSA) and methicillin-sensitive strain (MSSA) of Staphylococcus aureus MIC (μg / ml) MIC (μg / Ml) Strain No. Et ₂ O extract XS-1 Strain No. Et ₂ O extract XS-1 1 250 7.8 15 250 15.6 2 250 7.8 16 250 7.8 3 500 500 15.6 17 250 7.8 4 250 15.6 18 250 250 15.6 5 500 7.8 19 250 7.8 6 500 500 7.8 20 250 7.8 7 500 7.8 21 500 7.8 8 250 250 15.6 22 125 7.8 9 250 15.6 23 125 125 15.6 10 250 7.8 24 24 125 15.6 11 11 250 7.8 25 25 7.8 12 12 250 7.8 26 125 7.8 13 13 500 7.8 27 125 7. 14 250 15.6 shares No.1~20 is a methicillin-resistant strains of Staphylococcus aureus (MRSA), stock No.21~
27 is a methicillin-sensitive strain (MSSA) of Staphylococcus aureus.

Against Gram-positive and Gram-negative bacteria
Antibacterial activity of ether extract and XS-1 1. Staphylococcus epidermidis ( S.epiderm
idis ) 2. Bacillus cereus 3. Enterococcus faecalis (E.faecalis) 4. Acinetobacter calcoaceticus ( A. calcoacet
icus ) 5. C. freundii 6. Enterobacter cloacae 7. E. coli 8. Klebsiella Nyumonii (K.pneumoniae) 9. Pseudomonas aeruginosa (P.aeruginosa) 10.-flops Roteusu-Mirahi Bu squirrel (P.mirabilis) 11. Proteus vulgaris 12. S. marcescens 13. Salmonella Tifi (Sal.typhi) 14. Salmonella typhimurium (Sal.typhimuriu
m )

[0011]

[Table 2] Strain Et ₂ O-extract extract XS-1 1 125 31.3 2 125 62.5 3> 250> 250 4> 250> 250 5> 250> 250 6> 250> 250 7> 250> 250 8 125 31.3 9 250 125 10> 250> 250 11> 250> 250 12> 250> 250 13> 250 12514>250> 250

Results Xantatin was found to have antibacterial activity in all 20 strains of MRSA and 7 strains of MSSA, and the MIC value was 7.8 to 15.
It was 6 μg / ml (Table 1). In addition, a gram-positive bacterium Staphylococcus epidermidis (S.epidermidis
IFO3762), B. cereus IF
O3514) also had antibacterial activity, and the MIC values were 31.3 and 62.5 μg / ml, respectively. The antibacterial activity in Enterococcus faecalis (E.faecalis ATCC21212) was not observed. Almost no antibacterial activity was observed against Gram-negative bacteria (Table 2). From the above, it was clarified that xanthatin has antibacterial activity against some Gram-positive bacteria, particularly against Staphylococcus.

Effective Dosage and Administration Method Xantatin can be prepared as an internal medicine or an external preparation having an MRSA antibacterial effect. When used as an external preparation, an antibacterial agent or bactericidal agent for xanthatin is prepared using a conventional method, but by adding it to a commercially available disinfectant solution, for example, cresol water, cresol soap solution, disinfecting phenol water, etc. Can be used. These antibacterial agents or bactericides are used at a concentration of about 0.1 to 10 (weight or volume)% for disinfecting instruments, excrements of patients, and cleaning skin, mucous membranes and wounds.

When used as an internal preparation, it is used in the form of tablets, capsules, tablets coated with sugar, etc., liquid solutions or suspensions. The dose of the internal medicine varies depending on the age, body weight, symptom of the patient and administration route. For example, when administered to an adult (body weight of about 70 kg), 10 mg to 100 mg per daily administration Oral administration from 1 to 3 times.

The internal medicine is usually prepared according to a conventional method,
It is administered in a pharmaceutically suitable form. For example, solid oral forms may contain diluents such as lactose, dextrose, sucrose, cellulose, corn starch and potato starch, silica, talc, stearic acid, magnesium stearate or calcium stearate and / or polyethylene glycol together with the active compound. Lubricants, starches, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, polyvinylpyrrolidine and other binders, starch, alginic acid, alginates, disintegrating agents such as sodium starch glycolate, effervescent agents, dyes, sweeteners such as lecithin, polysorbate Humectants such as lauryl sulphate, and generally non-toxic and pharmaceutically inactive substances used in pharmaceutical formulations.

The above-mentioned internal medicine can be prepared by a known method such as mixing,
It is manufactured by granulation, tableting, sugar coating, coating method, or the like. Liquid dispersions for oral administration are eg syrups, emulsions, suspensions and solutions.

The syrup may contain saccharose as a carrier, for example with saccharose or glycerin and / or mannitol and / or sorbitol, especially the syrup administered to diabetics is one which is not metabolized to glucose as a carrier, Or it may contain only those that are metabolized to yield a very small amount of glucose, such as sorbitol. Suspensions and emulsions contain as carrier, for example, natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose or polyvinyl alcohol.

Solutions for oral administration may contain as carrier, for example, water, ethanol, propylene glycol, glycerin, saccharose and optionally pharmaceutically acceptable salts. Moreover, the best therapeutic effect can be obtained by changing the dose and administration route of these drugs and by using them in combination with other drugs having an MRSA antibacterial action.

[0019]

【Example】

Example 1 (Disinfectant) 5 g of xanthatin was dissolved in 1000 ml of normal water and used as a disinfectant.

Example 2 (tablet) According to a conventional method, 100 mg of xanthatin, 1 g of lactose, 300 mg of starch, 50 mg of methylcellulose and 30 mg of talc were prepared into 10 tablets and sugar-coated with sucrose.

Claims (1)

[Claims] 1. An anti-MRSA active pharmaceutical composition containing xanthatin as an active ingredient.