JPH09194358A - Pharmaceutical composition having anti-mrsa activity containing polyhydric phenol derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject composition capable of exhibiting an inhibitory activity against methicillin-resistant Staphylococcus aureus and free from a problem of severe side effect by compounding a polyhydric phenol derivative expressed by a specific formula as an active component. SOLUTION: This pharmaceutical composition contains a polyhydric phenol of formula I [R is a lower alkyl, OR1 (R1 is H or a lower alkyl) or (-) epigallocatechin anion] as an active component. Further, concretely, the derivative, especially a derivative expressed by formula II is preferably gallic acid, a gallate such as methyl gallate, ethyl gallate, n-propyl gallate or (-) epigallocatechin gallate, 2',3',4'-trihydroxyacetophenone or 2',4',5'- trihydroxybutyrophenone.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an anti-MRSA active pharmaceutical composition containing a polyhydric phenol derivative 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 among the ingredients in Chinese herbs that had no or no side effects, and found that gallic acid, which is an ingredient contained in Kashi And that there is anti-MRSA activity in polyphenol derivatives with similar structures.

Although gallic acid is widely present in leaves, stems, roots and the like of plants, a large amount of tannin is contained in koji, which is a kind of herbal medicine, and gallic acid is obtained by hydrolyzing this.訶子 is a genus (Cococidae) (Co
mbretaceae) Milobaran ( Terminalia chebula
R _ETZ .) Mature fruit. The primary plant is a deciduous tree, which blooms in summer and ties oval-shaped, brown, glossy fruits. When this is dried, 5 vertical lines are formed.
It has astringent, antidiarrheal, cough and hemostasis effects. Rinko is also used for skin tanning. However, it has never been reported that the silkworm has an anti-MRSA activity, and that gallic acid has an anti-MRSA activity. The present invention has been completed based on the above findings.

[0005]

The present invention provides an anti-MRSA active pharmaceutical composition containing a polyhydric phenol derivative as an active ingredient.

Specifically, the formula:

Embedded image [Wherein R is lower alkyl, OR ₁ (wherein R ₁ is H
Or a lower alkyl) or (−) is an epigallocatechin anion], which is an anti-MRSA active pharmaceutical composition containing as an active ingredient a polyhydric phenol derivative. Has the formula:

Embedded image [Wherein R is OR ₁ (wherein R ₁ is H or lower alkyl) or (−) epigallocatechin anion] and a polyhydric phenol derivative represented by the formula:

[Chemical 6] [Wherein R is lower alkyl] and is a polyhydric phenol derivative.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION As used herein, "lower alkyl" is a saturated straight chain or branched chain having 1 to 1 carbon atoms.
It refers to a hydrocarbon residue containing 6, preferably 1 to 5, and more preferably 1 to 4. For example, methyl, ethyl, n
-Propyl, isopropyl, butyl, t-butyl. Also, the 2, 5 and 6 positions of the benzene ring can be selected-
OH is preferably in the 5 position when R is OR ₁ (wherein R ₁ is H or lower alkyl) or (−) epigallocatechin anion, and when R is lower alkyl Is preferably in the 2 or 6 position.

The polyhydric phenol derivative of the present invention is preferably gallic acid, methyl gallate, ethyl gallate, n-propyl gallate, (−) epigallocatechin gallate,
2 ', 3', 4'-trihydroxy-acetophenone and 2 ', 4', 5'-trihydroxy-butyrophenone.

The polyhydric phenol derivative of the present invention generally exhibits substantially the same physiological activity or pharmacological activity as the free form in vivo, for example, the derivative of the compound of the present invention and a pharmaceutically acceptable salt thereof. , Addition salts, hydrates, etc. are included in the technical scope of the present invention.

Purification of gallic acid Crushed citrus fruit (Terminalia chebula) fruit (5 kg)
The mixture was refluxed with 0% ethanol for 2 hours, suction filtered while warm, and the filtrate was concentrated under reduced pressure. Ether was added to the concentrated ethanol extract, and liquid-liquid partition extraction was performed. Ether layer 70 ℃
The mixture was concentrated under reduced pressure to dryness under reduced pressure to give a brown viscous substance (67.73 g). This was dissolved in absolute ethanol and used as an ether extraction fraction. The water layer was subjected to liquid-liquid partition extraction by further adding water-saturated n-butanol, and the obtained n-butanol layer was concentrated to dryness under reduced pressure and dissolved in anhydrous ethanol to obtain an n-butanol extraction fraction. The aqueous layer fraction was concentrated under reduced pressure, evaporated to dryness and dissolved in water to give a water-extracted fraction. The ether extract was layered on a glass column (50 × 650 mm) packed with silica gel (Wakogel C-200) and fractionated with n-hexane-ethyl acetate to obtain a fraction having antibacterial activity. These active fractions were mixed with a glass column (4) packed with ODS-Q3 (wako).
(0 × 75 mm), fractionated with 50% methanol, and further purified with Develosil ODS 10 (20 × 250 mm) and purified with 35% methanol. As a result, ethyl gallate (TC-1) 1143 mg, gallic acid (TC-2) 8
Isolated in a yield of 63 mg.

¹ H-NMR spectrum data position in TC-1 and TC-2 TC-1 TC-2 2,6 7.05 (2H, s) 7.09 (s) CH ₂ 4.24 (2H, q , J = 7.0) CH ₃ 1.32 (3H, t, J = 7.0)

¹³ C-NMR spectrum data position in TC-1 and TC-2 TC-1 TC- 21 1 121.53 122.00 2,6 110.05 110.35 3,5 146.50 146.443 4 139.72 139.63 CO 168.59 170.43 CH ₂ 61.70 CH ₃ 14.66 Note that gallic acid may be a commercially available one.

The lower alkyl ester of gallic acid such as methyl gallate, ethyl gallate, n-propyl gallate, etc. is prepared by esterifying gallic acid according to a conventional method,
Alternatively, a commercially available product is used. Commercially available products of (−) epigallocatechin gallate, 2 ′, 4 ′, 5′-trihydroxyacetophenone and 2 ′, 3 ′, 4′-trihydroxybutyrophenone can also be used.

Antibacterial activity against MRSA As test bacteria, 20 strains of MRSA (methicillin resistant strain), M
Seven SSA (methicillin-sensitive strains), three other Gram-positive strains, and 11 Gram-negative strains were used. The antibacterial activity against MRSA, which was used as an index for isolation and extraction of the active ingredient, was measured by the disk diffusion method. MRSA bacterial solution about 1
0 ⁵ CFU (colony homing unit: colony formi
ng unit) is smeared on an agar plate and various concentrations of test sample 1
Place a disk (diameter 13 mm) that absorbed 00 μl and
After culturing at 7 ° C. for 20 hours, the presence or absence of inhibition zone formation was confirmed.
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.

Results MRSA20 strain, MSSA7 of ether extract fraction of licorice
The value of MIC for the strain was 31.3 to 250 μg / ml, whereas MI of gallic acid (TC-2: Gallic Acid) and ethyl gallate (TC-1: Ethyl Gallate) was determined.
C values are 62.5 to 125 μg / ml and 7.9 to 62, respectively.
It was 5 μg / ml and showed about twice as much antibacterial activity (see Table 1).

[0016]

Table 1 Antibacterial activity of Et ₂ O extract, TC-1 and TC-2 against methicillin resistant and methicillin sensitive strains of Staphylococcus aureus MIC (μg / ml) MIC (μg / ml) strain No. Et ₂ O extract extract TC-1 TC-2 strain No. Et ₂ O extract extract TC-1 TC-2 1 125 62.5 31.3 15 62.5 62.5 62.5 2 125 62.5 31.3 16 250 62.5 62.5 3 125 62.5 31.3 17 250 125 31.3 4 125 62.5 31.3 18 250 125 31.3 5 125 62.5 62.5 19 250 125 62.5 6 125 62.5 62.5 20 250 62.5 31.3 7 62.5 62.5 31.3 21 125 62.5 31.3 8 125 125 62.5 22 31.3 62.5 7.9 9 125 62.5 31.3 23 31.3 62.5 15.7 10 62.5 62.5 62.5 24 62.5 62.5 15.7 11 125 125 62.5 25 31.3 31.3 7.9 12 125 125 62.5 26 31.3 31.3 7.9 13 125 125 62.5 27 62.5 62.5 15.7 14 125 62.5 31.3 Strain Nos. 1 to 20 are methicillin resistant strains and strain No. 21
27 are methicillin sensitive strains.

The antibacterial activity against other 3 Gram-positive bacteria and 9 Gram-negative bacteria is TC-2 (Gallic Acid).
Neither TC-1 (Ethyl Gallate) showed particularly strong antibacterial activity against Gram-positive bacteria. For gram-negative bacteria, ethyl gallate showed slightly stronger activity than gallic acid, but not so strong activity (see Table 2).

1. Staphylococcus epidermidis 2. Bacillus cereus 3. Enterococcus faecalis 4. Acinetobacter calcoaceticus (A. calcoace
ticus) 5. C. freundii 6. E. cloacae 7. E. coli 8. K. pneumoniae 9. P. aeruginosa 10. P. aeruginosa Proteus Mirabilis 11. Proteus vulgaris 12. S. marcescens

[Table 2] Antibacterial activity of ether extract, TC-1 and TC-2 against Gram-positive and Gram-negative bacteria MIC (μg / ml) strain Et ₂ O-extract extract TC-1 TC-2 Gram positive 1. Philococcus epidermidis> 500 500 500 2. Bacillus cereus 250 500 500 3. Enterococcus faecalis> 500 1000 1000 Gram negative 4. Acinetobacter calcoaceticus 500 250 250 5. Citrobacter Freundii 500 250> 1000 6. Enterobacter Kuroake> 500 250> 1000 7. Escherichia coli 500 250> 1000 8. Klebzira pneumonia 250 250 250 9. Pseudomonas aeruginosa 500 250 1000 10. Proteus mirabilis 500 250 250 11. Proteus vulgaris 250 500 250 12. Serratia Marcessens 250 500 250

Next, the relationship between the chemical structure of gallic acid and the antibacterial activity was examined. N-Propyl gallate (n-Propyl Gallat), which is substituted for the carbornyl group of gallic acid.
e), methyl gallate, Lauryl gallate, (−) epigallocatechin gallate ((−) Epigallocatechin Gallate), one hydroxyl group is changed or 2 ′, 4 ′, 5'-trihydroxy-butyrophenone (2 ', 4', 5'-Trihydroxy-butyro
phenone), 2 ', 3', 4'-trihydroxy-acetophenone (2 ', 3', 4'-Trihydroxy-acetophenone), gallic acid trimethyl ether (Trimethylether Gallate) with the hydroxyl group changed to a methyl group, the number of hydroxyl groups 5-hydroxyoxyphthalic acid (5-Hydroxyisophthalic
Acid) 2,5-dihydroxybenzoic acid (2,5-Dih
The antibacterial activity of ydroxybenzoic acid was measured.

[0020]

Embedded image

Results: Epigallocatechin gallate substituted with a carbornyl group,
Although n-propyl gallate and methyl gallate showed MRSA antibacterial activity, lauryl gallate did not. 2 ', 4', 5'-trihydroxy-butyrophenone and 2 ', 3', 4'-trihydroxy-acetophenone having different hydroxyl positions showed strong activity. In addition, gallic acid trimethyl ether, 5-hydroxyisophthalic acid,
None of 2,5-dihydroxybenzoic acid showed antibacterial activity (see Table 3). In addition, these compounds did not show strong antibacterial activity against 2 Gram-positive strains and 9 Gram-negative strains (see Table 4).

[0022]

[Table 3] MIC (μg / ml) strain No. 1 62.5 62.5 62.5 31.3 31.3 2 62.5 62.5 62.5 31.3 31.3 3 62.5 62.5 62.5 31.3 31.3 4 62.5 31.3 62.5 31.3 31.3 5 62.5 62.5 125 31.3 31.3 6 62.5 62.5 62.5 31.3 31.3 7 62.5 62.5 31.3 31.3 31.3 8 62.5 62.5 31.3 31.3 31.3 9 62.5 31.3 62.5 31.3 31.3 10 31.3 62.5 62.5 31.3 31.3 11 62.5 62.5 125 62.5 62.5 12 62.5 62.5 125 62.5 62.5 13 62.5 62.5 125 62.5 31.3 14 62.5 62.5 62.5 62.5 31.3 15 62.5 31.3 62.5 31.3 31.3 16 62.5 62.5 62.5 62.5 31.3 17 62.5 62.5 125 62.5 62.5 18 31.3 62.5 62.5 31.3 31.3 19 62.5 62.5 62.5 62.5 31.3 20 62.5 31.3 125 62.5 62.5 21 31.3 62.5 62.5 31.3 31.3 22 15.7 31.3 62.5 31.3 7.9 231 11.3 31.3 31.3 31.3 15.7 24 15.7 31.3 31.3 31.3 15.7 25 3.9 31.3 31.3 3.9 7.9 26 26 3.9 31.3 31.3 3.9 7.9 27 15.7 31.3 31.3 31.3 15.7 Strain Nos. 1 to 20 are methicillin resistant strains and strain No. 21
27 are methicillin sensitive strains.

[0023]

[Table 4] The strain name is the same as in Table 2. MIC (μg / ml) strain Gram positive 1. Staphylococcus epidemidice 250 500 1000 125 250 2. Enterococcus faecalis 500 500> 1000 250 250 Gram negative 4. Acinetobacter calcoaceticus 250 250 250 62.5 62.5 5. Citrobacterium Freundy 250 250 250 62.5 125 6. Enterobacter Croake 500 500 250 62.5 250 7. Escherichia coli 1000 250 250 250 500 8. Crevella Newmonia 250 250 250 62.5 62.5 9. Shoot Monas Aergenosa 250 500 250 62.5 62.5 10. Proteus Mirabilis 250 250 500 62.5 125 11. Proteus vulgaris 125 125 250 62.5 62.5 12. Serratia Marcescens 250 50
0 500 62.5 62.5

From the above, gallic acid and its similar compounds, methyl gallate, ethyl gallate, and gallic acid n-
Propyl, epigallocatechin gallate, 2 ', 4', 5 '
-Trihydroxy-butyrophenone and 2 ', 3', 4 '
-Trihydroxy-acetophenone was found to have strong antibacterial activity against Staphylococcus aureus

Effective dose and administration method The polyhydric phenol derivative of the present invention can be prepared as an external preparation having an MRSA antibacterial action. The antibacterial agent or bactericidal agent for the polyhydric phenol derivative of the present invention is prepared by a conventional method, but by adding it to a commercially available disinfecting 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 disinfection of instruments, patient's excretion, and cleaning of skin, mucous membranes and wounds.

The polyphenol derivative of the present invention can be orally administered in various dosage forms. For example, in the form of tablets, capsules, tablets coated with sugar and the like, liquid solutions or suspensions.

The dose can be varied depending on the age, body weight, symptom of patient and administration route. For example, when administered to an adult (body weight of about 70 kg), 10 mg to 100 mg per administration Oral administration once to three times daily. The best therapeutic effect is obtained by changing the dose and administration route.

The pharmaceutical composition of the present invention is usually prepared by a conventional method and administered in a pharmaceutically suitable form. For example, solid oral forms include lactose, along with the active compound,
Diluents such as dextrose, saccharose, cellulose, corn starch and potato starch, silica,
Lubricants such as talc, stearic acid, magnesium stearate or calcium stearate and / or polyethylene glycol, binders such as starch, acacia, gelatin, methylcellulose, carboxymethylcellulose, polyvinylpyrrolidine, starch, alginic acid, alginates, glycolic acid Disintegrating agents such as sodium starch, effervescent agents, pigments, sweeteners, wetting agents such as lecithin, polysorbate, and lauryl sulfate,
And generally may include non-toxic and pharmaceutically non-active substances used in pharmaceutical formulations.

The abovementioned pharmaceutical preparations are prepared by known methods, for example by mixing, granulating, tabletting, sugar-coating or coating methods. 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 those syrups administered to diabetics are those which are 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 carriers, for example, water, ethanol, propylene glycol, glycerine, saccharose and optionally pharmaceutically acceptable salts.

[0032]

【Example】

Example 1 (Disinfectant) 0.5 g of gallic acid was dissolved in 1000 ml of normal water and used as a disinfectant.

Examples 2 to 7 (antiseptic solution) Instead of gallic acid, methyl gallate, ethyl gallate,
N-Propyl gallate, (−) epigallocatechin gallate, 2 ′, 3 ′, 4′-trihydroxy-acetophenone (all of which were purchased from Wako Pure Chemical Industries) and 2 ′, 4 ′,
A disinfecting solution was prepared in the same manner as in Example 1 using 5'-trihydroxy-butyrophenone (purchased from Kanto Kagaku).

Example 8 (tablet) By a conventional method, gallic acid 100 mg, lactose 1 g, starch 3
00mg, methylcellulose 50mg, talc 30mg 10
Prepare into tablets and sugar-coat with white sugar.

Examples 9 to 14 (tablets) Instead of gallic acid, methyl gallate, ethyl gallate,
Similar to Example 8 using n-propyl gallate, (−) epigallocatechin gallate, 2 ′, 4 ′, 5′-trihydroxy-butyrophenone and 2 ′, 3 ′, 4′-trihydroxy-acetophenone. To prepare tablets.

Example 15 (Syrup) To 0.05 g of gallic acid, 10 g of a single syrup was added and an appropriate amount of water was added to prepare a 50 ml syrup.

Examples 16 to 21 (Syrup) Instead of gallic acid, methyl gallate, ethyl gallate,
Example 15 using n-propyl gallate, (−) epigallocatechin gallate, 2 ′, 4 ′, 5′-trihydroxy-butyrophenone and 2 ′, 3 ′, 4′-trihydroxy-acetophenone. A syrup was prepared in the same manner.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location C07C 69/84 C07C 69/84 C07D 311/62 C07D 311/62 // C07C 49/825 9049-4H C07C 49/825

Claims (5)

[Claims] 1. The formula: embedded image [Wherein R is lower alkyl, OR ₁ (wherein R ₁ is H
Or a lower alkyl) or (−) epigallocatechin anion], and an anti-MRSA active pharmaceutical composition containing as an active ingredient a polyhydric phenol derivative. 2. The formula: [Wherein R is OR ₁ (wherein R ₁ is H or lower alkyl) or (−) epigallocatechin anion], and the anti-MRSA contains a polyhydric phenol derivative as an active ingredient. Active pharmaceutical composition. 3. The pharmaceutical composition according to claim 1, wherein the polyhydric phenol derivative is gallic acid, methyl gallate, ethyl gallate, n-propyl gallate or (−) epigallocatechin gallate. 4. The formula: An anti-MRSA active pharmaceutical composition comprising a polyhydric phenol derivative represented by the formula [wherein R is lower alkyl] as an active ingredient. 5. The polyhydric phenol derivative is 2 ′, 3 ′,
4'-trihydroxy-acetophenone or 2 ', 4',
5. It is 5'-trihydroxy-butyrophenone.
The pharmaceutical composition according to 4 above.