Quinolone carboxylic acid derivatives

Abstract

A quinolone carboxylic acid derivative having the following formula (1), ##STR1## wherein R.sup.1 is a hydrogen atom, an alkyl group, an aralkyl group, an ester residual group which can be hydrolyzed in living bodies, R.sup.2 is a hydrogen atom or an amino group which may be substituted by one or two lower alkyl groups, X is a hydrogen atom or a halogen atom, Y is CH.sub.2, O, S, SO, SO.sub.2, or N--R.sup.3, wherein R.sup.3 is a hydrogen atom or a lower alkyl group, and Z is an oxygen atom or two hydrogen atoms; or a salt thereof; and an antimicrobial agent comprising the same. The compound exhibits a superior antimicrobial activity, especially against gram positive microorganisms, and is thus useful for the treatments and prevention of various infectious diseases in clinics.

Description

TECHNICAL FIELD

The present invention relates to a novel quinolone carboxylic acid derivative and a salt thereof which exhibit excellent antimicrobial activity against both Gram positive and Gram negative microorganisms.

BACKGROUND ART

Synthetic antimicrobial agents such as nalidixic acid, piromidic acid, and the like are known as drugs for curing infectious diseases caused by Gram negative microorganisms. They exhibit, however, only deficient effects on intractable diseases such as pseudomoniasis and the like.

On the other hand, quinolone carboxylic acid derivatives substituted with a fluorine atom at 6 position, such as norfloxacin, ofloxacin, and cyprofloxacin, or quinolone carboxylic acid derivatives substituted with a chlorine atom at 8 position have been developed (Japanese Patent Laid-open (ko-kai) Nos. 225181/1986, 90183/1984) and clinically used because of their strong antimicrobial activity.

These conventional synthetic antimicrobial agents had defects of insufficient absorptivity in a living body, providing only low bioavailability, and of a low antimicrobial activity against Gram positive microorganisms.

Therefore, development of antimicrobial agents having strong antimicrobial activity against both Gram positive and Gram negative microorganisms, including resistant bacteria, and superior absorptivity in living bodies has been desired.

In view of such a situation, the present inventors have synthesized a number of quinolone derivatives and studied their antimicrobial activity and absorptivity in a living body, and found that quinolone carboxylic acid derivatives of the following formula (1) and their salts exhibited antimicrobial activities against Gram positive microorganisms much stronger than conventional quinolone carboxylic acid derivatives, while maintaining their strong antimicrobial activities against Gram negative microorganisms, as well as excellent absorptivity. Such findings have led to the completion of the present invention.

DISCLOSURE OF THE INVENTION

The present invention provides a quinolone carboxylic acid derivative having the following formula (1), ##STR2## wherein R.sup.1 is a hydrogen atom, an alkyl group, an aralkyl group, an ester residual group or which can be hydrolyzed in living bodies, R.sup.2 is a hydrogen atom or an amino group which may be substituted by one or two lower alkyl groups, X is a hydrogen atom or a halogen atom, Y is CH.sub.2, O, S, SO, SO.sub.2, or N--R.sup.3, wherein R.sup.3 is a hydrogen atom or a lower alkyl group, and Z is an oxygen atom or two hydrogen atoms; or a salt thereof.

The present invention also provides an antimicrobial agent comprising a quinolone carboxylic acid derivative of formula (1) or a salt thereof as an effective component.

BEST MODE FOR CARRYING OUT THE INVENTION

Given as examples of groups represented by R.sup.1 in formula (1) are, as alkyl groups, linear or branched alkyl groups having 1-12 carbon atoms, e.g., methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, n-hexyl, and the like; as aralkyl groups, benzyl, phenyl ethyl, methyl benzyl, naphtyl methyl, and the like; and as ester residual groups which can be hydrolyzed in living bodies, alkanoyloxyalkyl, alkoxycarbonyloxyalkyl, carbamoylalkyl, alkoxyalkyl, and the like; specifically, acetoxymethyl, 1-acetoxyethyl, ethoxycarbonyloxymethyl, carbamoylmethyl, carbamoylethyl, methoxymethyl, methoxyethyl, and the like. Examples of amino groups which may be substituted by one or two lower alkyl groups, represented by R.sup.2, include amino, methylamino, ethylamino, isopropylamino, dimethylamino, diethylamino, diisopropylamino, and the like. Examples given of halogen atoms represented by X are chlorine, fluorine, bromine, iodine, and the like. Lower alkyl groups used in the present invention may be linear or branched alkyl groups having 1-5 carbon atoms, e.g., methyl, ethyl, i-propyl, sec-butyl, t-butyl, amyl, and the like.

When R.sup.2 in formula (1) is other than hydrogen, the carbon atom to which R.sup.2 is bonded is asymmetrical, so that there are optical isomers; R and S compounds, for compounds (1) of the present invention. The present invention includes both the optical isomers and the racemate.

As examples of the salts of the compounds of formula (1) of the present invention, salts of alkali metal, inorganic acid, organic acid, and the like are given, and more specifically, lithium salts, sodium salts, potassium salts, and the like, as salts of alkali metal; hydrochloride, sulfate, nitrate, hydrobromide, phosphate, and the like, as salts of inorganic acids; and acetate, fumarate, maleate, lactate, citrate, tartarate, malate, oxalate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, and the like, as salts of organic acid.

In the preparation of the compound of formula (1) of the present invention [Compounds (1a) and (1b)] Compound (2) and Compound (3), for example, are reacted according to the following reaction scheme to produce Compound (1a), and Compound (1a) is halogenated to produce Compound (1b). ##STR3## wherein X' is a halogen atom, and R.sup.1, R.sup.2, Y and Z are the same as those previously defined.

The raw material, Compound (2), is a known compound and can be prepared by the method described, for example, in Journal of Medicinal Chemistry, 31, 983 (1989). Compound (3) is also a known compound and can be prepared by the method described, for example, in Journal of American Chemical Society, 106, 630 (1984).

In order to prepare Compound (1a) from Compound (2) and Compound (3), 1 mol of Compound (2) is reacted with 1-5 mols of Compound (3) in a solvent such as acetonitrile, dimethylsulfoxide, or the like at room temperature to 100.degree. C. for 1-7 days. After the reaction, precipitate is collected by filtration and washed with a suitable solvent, e.g., methanol, chloroform, ether, etc., to obtain a crude product. The crude product is purified by silica gel column chromatography or by recrystallization to obtain Compound (1a).

The preparation of Compound (1b) from Compound (1a) can be carried out by a method described, for example, in Japanese Patent Laid-open 90183/1989; that is, 1 mol of Compound (1a) is reacted with 1-10 mols of a halogenizing agent, such as sulfuryl chloride, chlorine, bromine, iodine, N-chlorosuccinic acid imide, N-bromosuccinic acid imide, or the like, in a suitable solvent, such as chloroform, dichloromethane, acetic acid, methanol, ethanol, or the like, at 0.degree.-100.degree. C. for 1-48 hours. After the reaction, water is added to collect precipitate by filtration and the precipitate is washed with water to produce a crude product. This crude product is purified by silica gel column chromatography or by the recrystallization to obtain Compound (1b).

If necessary, Compound (1) thus obtained is converted into a salt, such as a salt of alkali metal, inorganic acid, organic acid, or the like, according to a conventional method.

When Compound (1) of this invention thus prepared is used as an antimicrobial agent, it is administered orally at a dose of 200 to 800 mg per day or parenterally at a dose of 5 to 40 mg per day, depending on the weight, the age, the sex, the physical conditions, or the symptom of the patient or the manner of administration.

Compound (1) can be formed into various antimicrobial preparations, such as tablets, granules, powders, capsules, suspensions, injections, suppositories, or the like, according to conventional methods. When solid preparations are produced, compound (1) is mixed with excipients, and as required, with binders, disintegrators, lubricants, coloring agents, sweetening agents, flavoring agents, fillers, coating agents, sugar-coating agents, and the like, and formed into preparations such as tablets, granules, powders, capsules, suppositories, or the like according to known methods. When compound (1) is made into a preparation for injection, it is dissolved, suspended, or emulsified into an aqueous medium such as distilled water, or made into powder which is dissolvable when it is injected. Intravenous, intraarterial, intraportal, intraperitoneal, intramuscular, or subcutaneous injection are applicable.

EXAMPLES

The present invention is hereinafter described in more detail by way of examples and test examples, which are not intended to be limiting thereof.

EXAMPLE 1

1-Cyclopropyl-6-fluoro-7-(2,3,4,5,6,7-hexahydro-1H-1,4-diazepin-5-one-1-yl)-4-oxoquinoline-3-carboxylic acid (Compound No. 1)

0.265 g of 1-cyclopropyl-6,7-difluoro-4-oxoquinoline-3-carboxylic acid and 0.200 g of 2,3,4,5,6,7-hexahydro-1H-1,4-diazepin-5-one dissolved into 20 ml of acetonitrile and refluxed for 3 days. After cooling, the precipitate was collected by filtration, washed with chloroform, methanol, and ether in this order to obtain 0.300 g of colorless crystals of the target compound (yield: 83%).

IR.nu..sub.max.sup.KBr cm.sup.-1 : 1720, 1660, 1620 .sup.1 H-NHR .delta.ppm(NaOD+D.sub.2 O): 0.76-1.48(m, 4H), 2.50-3.80(m, 9H), 7.10(d, J=7.7Hz, 1H), 7.56(d, J=14.1Hz, 1H), 8.48(s, 1H)

m.p.: 300.degree. C. or above

EXAMPLE 2

8-Chloro-1-cyclopropyl-6-fluoro-7-(2,3,4,5,6,7-hexahydro-1H-1,4-diazepin-5-one-1-yl)-4-oxoquinoline-3-carboxylic acid (Compound No. 2)

0.84 ml of sulfuryl chloride was added dropwise to a suspension of 1.50 g of 1-cyclopropyl-6-fluoro-7-(2,3,4,5,6,7-hexahydro-1H-1,4-diazepin-5-one-1-yl)-4-oxoquinoline-3-carboxylic acid in 30 ml of chloroform, and the mixture was stirred for 12 hours at room temperature. After an addition of ice water, the precipitate was collected by filtration, washed with water, dried in air, and recrystallized in ethanol-chloroform to obtain 0.87 g pale yellow crystals of the target compound (yield: 53.0%).

IR.nu..sub.max.sup.KBr cm.sup.-1 : 1720, 1660, 1620 .sup.1 H-NHR .delta.(CD.sub.3 OD+CDCl.sub.3): 0.80-1.50(m, 4H), 2.50-3.00(m, 2H), 3.30-3.64(m, 7H), 7.52(s, 1H), 8.10(d, J=11.6Hz, 1H), 9.02(s, 1H)

m.p.: 282.degree.-284.degree. C. (decomposed)

EXAMPLES 3-9

Compound Nos. 3-9 listed in Table 1 were prepared in the same manner as in Example 1. Physicochemical data of these compounds are given in Table 2.

TABLE 1______________________________________ ##STR4##Compound No. A______________________________________ ##STR5##4 ##STR6##5 ##STR7##6 ##STR8##7 ##STR9##8 ##STR10##9 ##STR11##______________________________________ TABLE 2__________________________________________________________________________Compound No. Melting Point (.degree.C.), Property IR (KBr, cm.sup.-1) .sup.1 H-NMR (.delta. ppm)__________________________________________________________________________3 268-271 (decomposed) light yellow powder 1720, 1650, 1625 1.10-1.60(m, 4H), 2.80-3.15(m, 5H), 3.40-3.85(m, 7H), 7.44(d, J=7.7Hz, 1H), 7.98(d, J=14.1Hz, 1H), 8.76(s, 1H). (CDCl.sub.3 + CD.sub.3 OD).4 205-209 colorless powder 1630 1.00-2.20(m, 10H), 3.00-4.00(m, 6H), 7.28(d, J=9.0Hz, 1H), 7.96(d, J=15.4Hz, 1H), 8.76(s. 1H). (CDCl.sub.3 + CD.sub.3 OD).5 228-230.5 light yellow powder 1720, 1620 1.10-1.40(m, 4H), 1.40-2.10(m, 8H), 3.05-3.30(m, 1H), 3.30-3.80(m, 4H), 7.18(d, J=9.0Hz, 1H), 7.90(d, J=15.4Hz, 1H), 8.70(s, 1H). (CDCl.sub.3).6 184-187 light yellow powder 1720, 1625 1.00-1.65(m, 4H). 1.65-2.20(m, 6H), 2.32(s, 6H), 2.68-3.05(m, 1H), 3.10-3.75(m, 4H), 3.75-4.05(m, 1H), 7.25(d, J=9.6Hz, 1H), 7.90(d, J=15.4Hz, 1H), 8.68 (s, 1H). (CDCl.sub.3).7 199-203 (decomposed) light brown powder 1720, 1610 1.00-1.40(m, 4H), 2.50-3.10(m, 4H), 3.10-4.00(m, 10H), 7.39(d. J=7.9Hz, 1H), 7.71(d, J=15.2Hz. 1H), 8.54(s, 1H). (DMSO-d.sub.6).8 215-217 colorless powder 1625 1.00-2.20(m, 10H), 3.00-4.00(m, 6H), 7.28(d, J=7.7Hz, 1H), 7.96(d, J=15.4Hz, 1H), 8.75(s, 1H). (CDCl.sub.3 + CD.sub.3 OD).9 218-221 colorless powder 1725, 1620 1.00-2.20(m, 10H), 2.60-4.00(m, 6H), 7.25(d, J=7.7Hz, 1H), 7.85(d, J=15.4Hz, 1H), 8.75(s, 1H). (CDCl.sub.3 +CD.sub.3 OD).__________________________________________________________________________

EXAMPLES 10-16

Reactions were carried out in the same manner as in Example 2 to prepare Compound Nos. 10-16 listed in Table 3. Physicochemical data of these compounds are given in Table 4.

TABLE 3______________________________________ ##STR12##Compound No. A______________________________________10 ##STR13##11 ##STR14##12 ##STR15##13 ##STR16##14 ##STR17##15 ##STR18##16 ##STR19##______________________________________ TABLE 4__________________________________________________________________________Compound No. Melting Point (.degree.C.), Property IR (KBr, cm.sup.-1) .sup.1 H-NMR (.delta. ppm)__________________________________________________________________________10 136-140 light yellow powder 1720, 1640, 1610 0.80-1.50(m, 4H), 2.80-3.00(m, 2H), 3.08(s, 3H), 3.30-3.80(m, 6H), 4.20-4.50(m, 1H), 8.15(d, J=11.6Hz, 1H), 9.00(s, 1H). (CDCl.sub.3).11 275-278 (decomposed) colorless powder 1610-1640 0.55-2.30(m, 4H), 1.40-1.95(m, 6H), 2.75-3.45(m, 5H), 4.00-4.30(m, 1H), 7.90(d), J=11.6Hz, 1H), 8.75(s, 1H). (NaOD + D.sub.2 O).12 163-165 light yellow powder 1720, 1620 0.80-1.50(m, 4H), 1.55-2.00(m, 8H), 3.20-3.70(m, 4H), 4.24-4.55(m, 1H), 8.05(d, J=11.6Hz, 1H), 8.96(s, 1H). (CDCl.sub.3).13 210-215 (decomposed) light yellow powder 1720, 1610 1.00-1.45(m, 4H), 1.60-2.20(m, 6H), 2.60-2.80(m, 6H), 3.00-3.60(m, 4H), 3.80-4.00(m, 1H), 4.15-4.30(m, 1H), 8.15(d. J= 7.7Hz, 1H), 8.98(s, 1H). (CDCl.sub.3).14 202-208 (decomposed) brown powder 1723, 1630 0.90-1.30(m, 4H), 2.60-4.20(m, 13H), 4.20-4.50(m, 1H), 7.93(d, J=11.0Hz, 1H), 8.81(s, 1H). (DMSO-d.sub.6).15 255-257 (decomposed) colorless power 1600-1640 0.50-1.30(m, 4H), 1.40-2.00(m, 6H), 2.75-3.50(m, 5H), 4.00-4.30(m, 1H), 7.90(d, J=11.6Hz, 1H), 8.65(s, 1H). (NaOD + D.sub.2 O).16 241-244 (decomposed) colorless powder 1600-1640 0.50-1.30(m, 4H), 1.40-2.00(m, 6H), 2.80-3.50(m, 5H), 4.10-4.40(m, 1H), 7.90(d, J=11.6Hz, 1H), 8.65(s, 1H). (NaOD + D.sub.2 O).__________________________________________________________________________

EXAMPLES 17-21

Reactions were carried out in the same manner as in Example 1 or 2 to prepare Compound Nos. 17-21 listed in Table 5. Physicochemical data of these compounds are given in Table 6.

TABLE 5______________________________________ ##STR20##Compound No. A X.sup.1______________________________________17 ##STR21## H18 ##STR22## H19 ##STR23## H20 ##STR24## Cl21 ##STR25## Cl______________________________________ TABLE 6__________________________________________________________________________Compound No. Melting Point (.degree.C.), Property IR (KBr, cm.sup.-1) .sup.1 H-NMR (.delta. ppm)__________________________________________________________________________17 231-233 light yellow powder 1720, 1625 1.10-1.40(m, 4H), 1.80-2.20(m, 2H), 2.50-2.70(m, 2H), 2.80-3.10(m, 2H), 3.60-4.00(m, 5H), 7.33(d, J=7.9Hz, 1H), 7.80(d, J=15.4Hz, 1H), 8.58(s, 1H). (DMSO-d.sub.6).18 238-239 light brown powder 1712, 1625 1.10-1.40(m, 4H), 1.8-2.2(m, 2H), 3.6-4.0(m, 9H), 7.38(d, J=8.1Hz, 1H), 7.81(d, J=14.7Hz, 1H), 8.59(s, 1H). (DMSO-d.sub.6).19 colorless plate 0.60-1.20(m, 4H), 2.20-3.40(m, 9H), 7.10(d, J=8.0Hz, crystals 1H), 7.60(d, J=15.0Hz, 1H), 8.50(s, 1H). (DMSO-d.sub.6).20 yellow powder 0.80-1.40(m, 4H), 1.80-2.20(m, 2H), 2.80-4.20(m, 10H; 1H disappeared with D.sub.2 O); 8.09(d, J=10.8Hz, 1H), 8.93(s, 1H). (CDCl.sub.3).21 181-183 colorless needles 1726, 1602 0.80-1.40(m, 4H), 1.60-1.90(br, 1H; disappeared with D.sub.2 O), 1.90-2.30(m, 2H), 3.40-3.70(m, 4H), 3.70-4.10(m, 4H), 4.20-4.50(m, 1H), 8.03(d, J=11.0Hz. 1H), 8.91(s, 1H). (CDCl.sub.3).__________________________________________________________________________

TEST EXAMPLE 1

Antimicrobial Activity

Antimicrobial activities against bacteria listed in Tables 7-9 were measured according to the MIC measurement method of The Japan Chemotherapeutic Association. Ofloxacin was used as a control. The results are shown in Tables 7-9.

Medium: Mueller Hinton Medium

Sample dilution: A 1,000 mcg/ml solution in 25% dimethylsulfoxide was prepared. A series of solutions with various concentrations (100 mcg to 0.006 mcg) were prepared by successively diluting the solution with sterilized water by a factor of 1/2.

Amount of inoculated bacteria: 10.sup.6 /ml Cultivation conditions: 37.degree. C., 48 hours Determination: after 24 hours TABLE 7______________________________________ MIC (.mu.g/ml) Com- pound Compound NumberTestes Bacteria Number 2 15 Ofloxacin______________________________________Gram-positive bacteria1. Bacillus subtilis 0.024 0.024 0.098 ATCC 66332. Staphylococcus aureus 0.012 0.012 0.195 FDA 209P3. Staphylococcus aureus 0.098 0.195 0.78 Terajima4. Staphylococcus aureus 0.012 0.024 0.39 Smith5. Staphylococcus epidermidis 0.049 0.049 0.78 ATCC 122286. Sarcina lutea ATCC 9341 0.195 0.098 3.127. Strepto occus faecalis 0.195 0.195 1.56 IFO 129648. Micrococcus lysodeikticus 0.049 0.195 1.56 IFO 3333______________________________________ TABLE 8______________________________________ MIC (.mu.g/ml) Com- pound Compound NumberTested Bacteria Number 2 15 Ofloxacin______________________________________Gram-negative bacteria9. Escherichia coli 0-1 0.195 0.098 0.09810. Escherichia coli H-12 0.39 0.098 0.09811. Salmonella typhi TD 0.098 0.049 0.02412. Shigella flexneri 2b 0.003 0.006 0.01213. Pseudomonas aeruginosa 6.25 1.56 1.56 IFO 1373614. Pseudomonas aeruginosa P2 12.5 1.56 0.7815. Pseudomonas aeruginosa 6.25 0.78 3.12 IFO 1258216. Klebsiella pneumonias 0.012 0.012 0.024 ATCC 1003117. Klebsiella pneumonias 0.098 0.049 0.024 IFO 1354118. Proteus vulgaris OXK 0.024 0.049 0.04919. Proteus rettgeri 0.78 0.195 0.09820. Serratia marcescens NHL 0.78 0.195 0.049______________________________________ TABLE 9______________________________________ MIC (.mu.g/ml) Com- pound Compound NumberTested Bacteria Number 2 15 Ofloxacin______________________________________Metricillin-resistantstreptococcus aureusgram-positive bacteria21. M.R. Staphylococcus aureus 0.049 0.049 0.39 39522. M.R. Staphylococcus aureus 0.024 0.024 0.39 41523. M.R. Staphylococcus aureus 0.024 0.024 0.39 41924. M.R. Staphylococcus aureus 0.024 0.049 0.39 42025. M.R. Staphylococcus aureus 0.024 0.049 0.39 421______________________________________

INDUSTRIAL APPLICABILITY

The compound (1) and its salt of this invention exhibit a superior antimicrobial activity, especially against Gram positive microorganisms, and are thus useful for the treatments and prevention of various infectious diseases in clinics.

Claims

1. A compound having the formula (I), ##STR26## wherein Z is two hydrogen atoms, Y is CH.sub.2, S, SO or SO.sub.2, R.sup.1 is a hydrogen atom, an alkyl group, an aralkyl group, an ester residual group which can be hydrolyzed in living bodies, R.sup.2 is a hydrogen atom or an amino group which may be substituted by one or two lower alkyl groups, X is a hydrogen atom or a halogen atom; or a salt thereof.

2. The compound of claim 1, wherein Z is two hydrogen atoms, Y is CH.sub.2, S, SO or SO.sub.2, R.sup.1 is a hydrogen atom, R.sup.2 is an amino group, and X is a hydrogen atom or a halogen atom.

3. The compound of claim 1, wherein X is a chlorine atom.

4. The compound of claim 2, wherein X is a chlorine atom.

5. An antimicrobial composition effective against bacteria selected from the group consisting of Gram negative and Gram positive microorganisms, comprising an antimicrobially effective amount of a compound of formula (I), ##STR27## wherein Z is two hydrogen atoms, Y is CH.sub.2, S, SO or SO.sub.2, R.sup.1 is a hydrogen atom, an alkyl group, an aralkyl group, an ester residual group which can be hydrolyzed in living bodies, R.sup.2 is a hydrogen atom or an amino group which may be substituted by one or two lower alkyl groups, X is a hydrogen atom or a halogen atom; or a salt thereof, and a pharmaceutically acceptable carrier.

6. The composition of claim 5, wherein Z is two hydrogen atoms, Y is CH.sub.2, S, SO or SO.sub.2, R.sup.1 is a hydrogen atom, R.sup.2 is an amino group, and X is a hydrogen atom or a halogen atom.

7. The composition of claim 5, wherein X is a chlorine atom.

8. The composition of claim 6, wherein X is a chlorine atom.

9. A method for curing an infectious disease caused by a microorganism selected from the group consisting of Gram negative and Gram positive microorganisms, comprising administering to a patient in need thereof an effective amount of a compound of the formula (I), ##STR28## wherein Z is two hydrogen atoms, Y is CH.sub.2, S, SO or SO.sub.2, R.sup.1 is a hydrogen atom, an alkyl group, an aralkyl group, an ester residual group which can be hydrolyzed in living bodies, R.sup.2 is a hydrogen atom or an amino group which may be substituted by one or two lower alkyl groups, X is a hydrogen atom or a halogen atom; or a salt thereof.

10. The method of claim 9, wherein Z is two hydrogen atoms, Y is CH.sub.2, S, SO or SO.sub.2, R.sup.1 is a hydrogen atom, R.sup.2 is an amino group, and X is a hydrogen atom or a halogen atom.

11. The method of claim 9, wherein X is a chlorine atom.

12. The method of claim 10, wherein X is a chlorine atom.

13. The composition of claim 5, wherein said Gram positive microorganisms are methicillin-resistant.

14. The method of claim 9, wherein said Gram positive microorganisms are methicillin-resistant.