CA2235485A1 - Methods for decomposing quinolones and naphthyridones - Google Patents

Abstract

The invention concerns two methods of treating quinolones and naphthyridones having an antimicrobial, in particular antibacterial, action, as well as derivatives of such compounds with wood-rotting fungi, optionally in the presence of water and/or inert solid or liquid substrates and other auxiliaries.

Description

CA 0223~48~ 1998-04-21 Le A 31 372-Forei~n Countries/ Rt /ngb/S-P
FILE, P~l~ IN THIS AMEMD~Q
TRANSLATION

Process for the de~radation of quinolones and naphthyridones The present invention relates to two processes for the degradation of anti-5 microbially, in particular antibacterially, acting quinolone- and naphthyridone-carboxylic acids and derivatives thereof with the aid of wood rot fungi.

Antimicrobially acting substances are widely used in medicine and veterinary medicine. The preparation of these active compounds is accompanied by problems in the elimin~tion of productio:n residues, wastewaters and packaging containing10 the active compound. Wastewaters cont~ining active compound from high-volume users, such as hospitals, animal breeders and animal fatteners, also present problems. Further problems ma~ arise with the decont~min~tion of waters or soilsif active compound is undesirab]y released in the event of accidents.

In such cases, the antimicrobial activity of the active compounds can prevent the 15 customary degradation by bacteria in soils, waters and sewage treatment plants.
Under certain circumstances, an undesirable interference, for example in biologic-ally operating sewage treatment plants, may even occur if relatively large quantities of wastewaters containing active compound have to be processed, i.e. in all thesc- cases an expensive pretreatment of the wastewaters, residues and the like 20 is necessary.

A simp:le treatment with which the active compounds can be degraded to the extent that they lose their antibiotic activity and their further degradation can take place by the customary biological degradation methods should be found for these cases.

25 Processes for the degradation of organic harmful substances with the aid of lignin-degrading fungi were known from a) German Offenlegungsschrift 4 104 624 and b) EP 0 192 237 A1.

However, the degradation in a) is demonstrated only on the example of aromatic hydrocarbons. According to G-erman Offenlegungsschrift 4 l 04 624, halogen-30 containing hydrocarbons cannot be degraded in this manner.

EP 0 192 237 A1 mentions some examples of chlorinated hydrocarbons which canbe degraded by the white rot fungus Phanerochaete chrysosporium.

CA 0223~48~ 1998-04-21 Le A 31 372-Foreign Countries It was not to be expected that the fluorine-containing heteroaromatic quinolonesand na~phthyridones are attacked by white rot fungi and, in particular, brown rot fungi or even wood-degrading Ascomycetes. In contrast there is the conviction, expressed publicly many times, that there are no microorg~ni~m~ which can degrade such compounds (cf., for example, in the Swedish Journal "Land" June 1995, page 4). There is no indication in the scientific literature of a significant degradation of such compounds by microorg~ni.~m.~. Likewise, brown rot fungi generally are attributed only very limited capabilities in the cleavage of aromatic rings with furthermore significantly lower capacities (compared with white rot fungi; J. Buswell, 1991). In the abstracts of the "Division of Agrochemicals of the American Chemical Society 1993 issue No. 44", the extremely low bioavailability of l4C-labeled sarafloxacin in soil samples is reported in one abstract (No. 92). It was found that sarafloxacin was not notably degraded by microorg~ni~m.~ in the soil.

The present invention relates to:

1. A process for the treatment of antimicrobially, in particular antibacterially, acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof with wood rot fungi, in particular white rot or brown rot fungi, if appropriate in the presence of water and/or inert solid or liquid substrates and further auxiliaries.

2. A process for the degradation of antibacterially and antimicrobially acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof, characterized in that the active compounds are treated with wood rot fungi, in particular white or brown rot fungi, if appropriate in the presence of water and/or inert solid or liquid substances and further auxiliaries.

3. A process for the' treatment of wastewaters which comprise antibacterially and antimicrobially acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof -with wood rot fungi, in particular white or brown rot fungi.

30 4. A process for the treatment of manure from intensive animal husbandry, which comprises antibacterially and antimicrobially acting quinolone- and CA 0223~48~ 1998-04-21 Le A 3] 372-Forei~n Countries naphthyridonecarboxylic acids and derivatives thereof with wood rot fungi~
in particular white or brown rot fungi.

l~ process for the treatment of soils which comprise antibacterially and ~mtimicrobially acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof with wood rot fungi, in particular white or brown rot f'ungi.

The best known wood-destroying Basidiomycetes are divided into 2 large groups:
on the one hand into the lignin-degrading white rot fungi, and on the other handinto the cellulose- and hemicelhllose-degrading brown rot fungi, which essentially do not degrade lignin but merely modify it by hydroxylation and demethylations.
White rot fungi are distinguished by extracellular enzymes, such as lignin peroxidases, m~ng~nese peroxidases and laccases, which as a rule are missing from brown rot fungi. The two groups can be distinguished by chemical detection methods (cf., for example, Stalpers, 1978, Studies in Mycology, No. 16, CentralLbureau voor Schimmelcultures, Baarn, Holland). In the case of brown rot fungi, the hydroxyl radical is of decisive importance for the degradation activity.
The wicLe differences between white and brown rot fungi, in particular in respect of the molecular mech~nism~ ol the degradation of wood, are documented in the technical literature (cf., for example, Evans et al., FEMS Microbiologica]L Reviews 13 (199~) 235-240).

Examples which may be mentioned of genera of wood rot fungi which can be employed in the processes acconiing to the invention are:

White rot fun~i:

Bjerkandera sp.
25 Cladoderris sp.
Coriolopsis sp.
Coriolus sp.
Ganoderma sp.
Irpex sp.
30 Phaneroc,haete sp.
Phellinus sp.
Phlebia sp.

CA 0223~48~ 1998-04-21 Le A 31 372-Forei~n Countries Pleurotus sp .
Trametes sp.
Trichaptum sp.

Brown rot fun~i:

5 Fomitopsis sp.
Gloeophyllum sp.
Leatiporus sp.
Pholiota sp.
Serpula sp.

10 Litter de~raders:

Agrocybe sp.
Hypholc,ma sp.
Stropharia sp.

Others:

15 Xylaria 5p. (Ascomycet) The following species may be mentioned as preferred examples:

White rot fun~i:

Bjerkandera adusta DSM 4708 Cladoderris dendritica DSM 9611 20 Coriolopsis rigida DSM 9596 Coriolus sp.
Ganoderma lucidum DSM 9621 Irpex lacteus DSM 9595 Phanerochaete chrysosporium DSM 9620 25 Phanerochaete chrysosporium DSM 1556 Phanerochaete chrysosporium ATCC 24725 Phellinus gilvus DSM 9602 Phlebia radiata DSM 5111 CA 0223~48~ 1998-04-21 Le A 31 372-Forei~n Countries Pleurotus sp. DSM 9618 Trametes versicolor DSM 3086 Trametes villosa DSM 9589 Trichaptum bysogenum DSN 9:597 5 Brown rot fun~i:

Fomitopsis spraguei DSM 9600 Gloeophyllum sepiarium DSM 6420 Gloeophyllum striatum DSM 9594 Gloeophyllum striatum DSM 9592 10 Gloeophyllum striatum DSM 1()335 Gloeophyllum trabeum DSM 3()87 Leatiporus sulphureus DSM 2785 Pholiota aurivella DSM 5070 Serpula lacrymans DSM 8728 15 Litter de~raders:

Agrocybe sp.
Hypholoma sp.
Stropha,ria rugosoannulata DSM: 9616 Stropha,ria sp.

20 Others:

Xylaria digitata DSM 914 (Ascomycete) Those which may be mentioned as especially preferred are:

White :rot fun~i:

Irpex la,cteus DSM 9595 25 Phanerochaete chrysosporium DlSM 1556 Phanerochaete chrysosporium DSM 9620 CA 0223~48~ 1998-04-21 Le A 31 372-Forei~n Countries Brown rot fun~i:

Gloeophyllum striatum DSM 9';92 Gloeophyllum striatum DSM 9';94 Gloeophyllum striatum DSM 1()335 5 Litter de~raders:

Stropharia rugosoannulata DSM: 9616 Wood rot fungi are known. The abovementioned species of wood rot fungi are included in public collections of microorg~ni.~m~ and are freely accessible; forexample~ those marked DSM are accessible from the Deutsche Sammlung fur 10 Mikroorganismen [German Collection of Microorganisms] (DSMZ), Mascheroder Weg 1 b, 3 81 16 Braunschweig.

It is readily possible to replace one of the species mentioned by another from the same group.

Antibacterially and antimicrobially acting quinolone- and naphthyridonecarboxylic 15 acids which may be mentioned are the compounds of the following formulae (I) and (II)I

x2 o X'~, COOR2 Y A N (I) R' x2 o X'~,COOR2 J~ 1 3 (II) B~Z--R
in which Le A 31 372-Forei~n Countries Xl and x2 independently of one another represent hydrogen, halogen, Cl 4-alkyl, Cl 4-alkoxy or NH2, Y represents radicals of the structures R'~R6 4,Nf~ 4,N~J N~J

~N~ 1<~4 ~ N/~N

R

~\N [~N---lwherein Rl6 = E~, -CH3, -CH=CH-CO2Rl6, -CH2-CH2-CO2Rl6, -CH2-CO-CH3,-CH2-CH2-CN, Rl6l - Me or Et, ]D = O or -CH2-, Le A 31 372-Forei~n Countries R4 represents hydrogen, optionally hydroxyl- or methoxy-substituted straight-chain or branched C~-C4-alkyl, cyclo-propyl or acyl having 1 to 3 C atoms, Rs represents hydrogen, methyl, phenyl, thienyl or pyridyl7 :R6 represents hydrogen or Cl 4-alkyl, R7 represents hydrogen or Cl 4-alkyl, JR8 represents hydrogen or Cl 4-alkyl, ]R9 represents hydrogen, methyl or -CH2-NR7R~, ]Rl~ represents hydrogen, Cl 4-alkyl, amino, optionally hydroxyl-substituted alkyl- or dialkylarnino having 1 or 2 C atoms in the alkyl part, amino-CI 4-alkyl, C] 4-alkylamino-Cl 4-alyl or 1-imidazolyl, Rll represents hydrogen, hydroxyl, methoxy, methylthio, halogen, methyl or hydroxymethyl, R12 represents hydrogen or methyl, ]~13 represents hydrogen or Cl 4-alkyl, ]~14 represents hydrogen or Cl 4-alkyl and ]Rl5 represents hydrogen or Cl 4-alkyl, Rl represents hydrogen, an optionally halogen-substituted alkyl residue having 1 to 3 carbon atoms, cyclopropyl, methoxy, 4-fluorophenyl, 2,4-difluorophenyl or methylarnino, R2 represents hydrogen or optionally methoxy- or 2-methoxyethoxy-substituted alkyl having 1 to 6 carbon atoms, CA 0223.748.7 1 998 - 04 - 2 1 Le A 31 372-Foreign Countries as well as cyclohexyl, benzyl, 2-oxopropyl, phenacyl, ethoxycarbonylmethyl or pivaloyloxymethyl, R3 represents hydrogen, methyl or ethyl and Xl and Y, together wi.th the adjacent C atoms, represent a dioxolanyl residue or dioxoamyl residue, A represents nitrogen or =CH-, =C(halogen)-, = C(OCH3)-, =C(CH3)- or=C(CN)-, :B represents oxygen, optionally methyl- or phenyl-substituted =N- as well as =CH27 .Z represents =CH- or =N-, .and their pharmaceutically usable hydrates, acid addition salts and salts with bases. The compounds of the formulae I and II can be present in the :form of their racemates or in enantiomeric forms.

Preferred compounds of the formulae (I) and (II) are those 15 in which Rl represents optionally fluorine-substituted Cl-C3-alkyl, or represents cyclo-~propyl, R1 represents an alkyl residue having 1 to 3 carbon atoms, cyclopropyl, 2-fluoroethyl, methoxy, 4-fluorophenyl, 2,4-difluorophenyl or methylamino, R2 represen-ts hydrogen or optionally methoxy- or 2-methoxyethoxy- substituted alkyl having 1 to 6 carbon atoms as well ;lS cyclohexyl, benzyl, 2-oxopropyl, phenacyl, ethoxycarbonylmethyl or pivaloyloxymethyl, R3 represents hydrogen, methyl or ethyl and CA 0223=,48=, l998-04-2l Le A 31 372-Forei~n Countries A represents nitrogen or =CH-, =C(halogen)-, = C(OCH3)-, =C(CH3)- or=C(CN)-, :B represents oxygen, optionally methyl- or phenyl-substituted =N- as well as =CH2 and 'Z represents=CH- or=N-, XI and x2 independently of one another represent hydrogen, halogen or -~n~2~

Y represents radicals of the structures I
D

R4~J R4'Nf~ R4--Nf~ N~

~ ~J R1 ~[~/ N ~Z~>CN ~\N--wherein ]~16 = H or -C:H3, D = O or -C:H2-, CA 0223~48~ 1998-04-21 Le A 31 372-Foreign Countries R4 represents hydrogen, optionally hydroxyl-substituted straight-chain or branched C1-C3-alkyl or oxyalkyl having 1 to 4 C
atoms, R5 represents hydrogen, methyl or phenyl, R6 represents hydrogen or methyl, R7 represents hydrogen or methyl, R9 represents hydrogen, methyl or -CH2-NH2, Rl~ represents hydrogen, methyl, amino, methylamino, dimethyl-amino, aminomethyl, methylaminomethyl or ethylamino-1 0 methyl, Rl1 represents hydrogen, hydroxyl, methyl, fluorine, methyl or hydroxymethyl, R13 represents hydrogen or methyl, Rl4 represents hydrogen or methyl, R15 represents hydrogen or methyl, and their pharmaceutically usable hydrates and acid addition salts as well as their alkali metal, alkaline earth metal, silver and guanidinium salts.

Particularly preferred compounds of the formula (I) are those in which r R1 represents an alkyl radical having 1 to 3 carbon atoms, cyclopropyl, 2-fluoroethyl, methoxy, 2,4-difluorophenyl or methylamino, :R2 represents hydrogen or optionally methoxy- or 2-methoxyethoxy- substituted alkyl having 1 to 6 carbon Le A 31 372-Forei~n Countries atoms, as well as cyclohexyl, benzyl, 2-oxopropyl, phenacyl, ethoxycarbonylmethyl or pivaloyloxymethyl, A represents nitrogen or =CH-, =C(halogen)-, =C(CH3)-, =C(OCH3)- or=C(CN)-, Xl and x2 independently of one another represent hydrogen, fluorine or ~H2~

Y represents radicals of the structures J R4--Nf ~ R4--Nf ~J N~

~, Rl~ R 4 ~N ~\N
R R'5 I

~ N
'Dl/

~herein ~16 = H or -Cl~3, r~ = O or -C~2-, R4 represents :hydrogen, optionally hydroxyl-substituted straight-chain or branched Cl-C3-alkyl or hydroxyalkyl having 1 to 4 C atoms, CA 0223.748.7 1 998 - 04 - 2 1 Le A 31 372-Forei~n Countries Rs represents hydrogen, methyl or phenyl, R6 represents hydrogen or rnethyl, R7 represents hydrogen or methyl, R9 represents hydrogen, methyl or -CH2NH2, :RI~ represents hydrogen, methyl, amino, methylamino, dimethyl-amino, aminomethyl, methylaminomethyl or ethylamino-methyl, :RI I represents hydrogen, hydroxyl, methyl, fluorine, methyl or hydroxymethyl, RI3 represents hydrogen or methyl, ]Rl4 represents hydrogen or methyl, ]Rls represents hydrogen or methyl, and the-ir pharmaceutically usable hydrates and acid addition salts as well as their alkali rr~etal, alkaline earth metal, silver and guanidinium salts.

15 Active compounds which may be mentioned as especially preferred are those with the common names enroflo-xacin, ciprofloxacin, danofloxacin, ofloxacin, norfloxacin, benofloxacin, sarafloxacin, difloxacin, orbifloxacin, marbofloxacin and oxolinic acid.

Active compounds which may be mentioned in particular are enrofloxacin, ~0 ciprofloxacin, marbofloxacin, ofloxacin, norfloxacin and oxolinic acid.

The prc,cess according to the invention is preferably carried out in liquid, solid or semi-so:lid homogeneous or heterogeneous media.

CA 0223.748.7 1 998 - 04 - 2 1 Le A 31 372-Forei~n Countries In addil:ion to the active compound to be degraded and the wood rot fungus, these comprise lignin-, lignocellulose- and/or cellulose-containing material. This includes straw, chaff, wood, wood chips, sawdust and ground cereal residue.

The processes can be described as follows:

5 1. :Process for the degradation of quinolone dissolved in water The preculture of one of the wood rot fungi listed which has been cultured ~ in malt broth which comprises malt extract or an equivalent source of carbon and energy in a concentration of 1-50 g/l, preferably 5-30 g/l, in particular 5-8 g/l, - at 5-40~C, preferably 15-30~C, in particular at 18-22~C, ~ while standing or being shaken, preferably shaken at 80 -120 rpm, - for a period of 3-28 days, preferably 5-14 days, in particular 5-7 days, ] 5 - in the dark, in daylight or in all intermediate stages of illumination, is added to a rinsing water, for example from the preparation process for a quinolone-contAining medicament, having a content of 0.01 to 500 ppm, preferably 0.1 to 50 ppm, in particular 1 to 20 ppm, of the quinolone in a ratio of 0.01 to 100 g of dry weight DWtl, preferably 0.1 to 10 g of DW/l, in particular 1 to ~ g of DW/I.

The pH of the suspension can be 3 to 8, preferably 4 to 7, in particular 5 to 6; phosphate is not added. Organic acids, such as, for example, acetic acid or oxalic acid, can be used to adjust the pH.

The content of iron2+ or iron3+ should be 0.1 to 100 IlM (10-6 mol per liter), preferably 1 to 30 ~M, in particular 2 to 20 ~M, CA 0223~48~ 1998-04-21 Le A 31 372-Forei~n Countries the content of NH4+ can be 0 001 to 20 mM (10-3 mol per liter), preferably - 0.1 to 10 mM, in particular between 0.1 and 1 mM.

The content of divalent cations should be between 0.1 and 10 mM, preferably 1 mM (for example 0.2 mM CaC12 + 0.8 mM MgSO4).

S This suspension is incubated under the same conditions as the preculture.

~he addition of coniferyl or veratryl alcohol in a concentration of 0.01 to 10 mM, preferably 0.1 to 3 mM, in particular 0.5 to 1 mM, results in an increase in the degrading activity for the quinolone.

2. :Process for degrading quinolone bounded to straw, manure or soil Starting from stock cultures of wood rot fungi on Petri dishes with malt agar, lignocellulose-contiining substances, in particular straw, sawdust or wood chips, are inoculated with agar blocks punched out of the stock culture, 0.1 to 100 g of agar per kg of solid substrate, preferably 1 to 10 g of agar per kg of substrate, being employed.

Before the inoculation, water is added to the solid matrix (straw, manure, soil, sludge) comprising the quinolone (0.1 to 200 ppm) until a water content of 10 to 90%, preferably 40 to 50% (based on the weight), is r eached.

During the inoculation, the inoculum is distributed in the solid matrix. The subsequent incubation is carried out at 5 to 40~C, preferably 15 to 30~C, in particular at 18 to 22~C.

CA 0223~48~ 1998-04-21 Le A 31 372-Forei~n Countries The following examples illustrate the processes according to the invention:

Examples Example 1 One liter of rinsing water from a formulating tank having a content of 10 ppm of5 enrofloxacin is inoculated with a preculture of Gloeophyllum striatum DSM 9592or Gloeophyllum striatum DSM 10335, during which the fungus mycelium is to be transferred from the preculture in 1/4 concentrated malt broth into the rinsing water such that a concentration of 2 g of dry weight per liter is achieved. Thissuspension is incubated at room temperature while shaking at 100 rpm in the dark.
After 2 weeks, 10 to 20% of the [carbonyl-l4C]-enrofloxacin employed was degraded to l4co2.

Example 2 1 kg of wheat straw which comprises 2 mg of enrofloxacin bonded adsorptively was moistened with 1 kg of water and inoculated with Gloeophyllum striatum DSM 9594. The inoculum comprises 20 agar blocks each weighing about 0.5 g.
These small blocks were punched out of a preculture from the fungal colony on malt agar in a Petri dish. The batch was incubated at room temperature in the dark while gassing with moistened air. After 8 weeks, 50% of the enrofloxacin was degraded.

20 Example 3 0.5 kg of moistened wood chips were inoculated with Phanerochaeta chrysosporium DSM 9620 and preincubated for 1 week at room temperature. 0.5 kg of manure from an anlmal population which had been treated with enrofloxacin was adcled to this batch. The enrofloxacin content in the manure was 2 ppm. The 25 batch was incubated at room temperature.

Example 4 0.1 kg of wheat straw was moistened with 100 ml of water and inoculated with Gloeophyllum striatum DSM 9594. After preincubation for 7 days at room Le A 31 372-Forei~n Countries temperature, the straw was covered with a layer of 0 6 kg of field soil (with a water content of about 50%) comprising 10 ppm of enrofloxacin and was incubated at room temperature. After 15 weeks, 6% of the enrofloxacin employed was degraded.

Claims (9)

claims

1. Process for the treatment of antimicrobially, in particular antibacterially, acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof with wood rot fungi, if appropriate in the presence of water and/or inert solid or liquid substrates and further auxiliaries.

2. Process for the degradation of antimicrobially, in particular antibacterially, acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof, characterized in that the active compounds are treated with wood rot fungi, if appropriate in the presence of water and/or inert solid or liquid substrates and further auxiliaries.

3. Process for the treatment of wastewaters which comprise antimicrobially, in particular antibacterially, acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof with wood rot fungi.

4. Process for the treatment of manure from intensive animal husbandry which comprises antimicrobially, in particular antibacterially, acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof with wood rot fungi.

5. Process for the treatment of soils which comprise antimicrobially, in particular antibacterially, acting quinolone- and naphthyridonecarboxylic acids and derivatives thereof with wood rot fungi.

6. Process according to Claims 1 to 5, characterized in that the active compound is enrofloxacin or ciprofloxacin.

7. Process according to Claims 1-6, characterized in that white rot fungi are employed for the degradation.

8. Process according to Claims 1-6, characterized in that brown rot fungi are employed for the degradation.

9. Process according to Claims 1-6, characterized in that wood-degrading Ascomycetes are employed.