JP2005532358A - Peptide deformylase inhibitor - Google Patents

Abstract

The present invention relates to a novel PDF inhibitor and a novel method for its use.

Description

Detailed Description of the Invention

(Field of Invention)
The present invention relates to the use of novel N- [2- (N-formyl-N-hydroxy-amino) -ethyl] -arylamide compounds and pharmaceutical compositions comprising these compounds as peptide deformylase inhibitors.

(Background of the Invention)
Bacterial initiator methionyl tRNA is modified by methionyl tRNA formyltransferase (FMT) to produce formyl-methionyl tRNA. The formylmethionine (f-Met) is then incorporated into the N-terminus of the newly synthesized polypeptide. Polypeptide deformylase (PDF or Def) then deformylizes the primary translation product to yield the N-methionyl polypeptide. Most intracellular proteins are further processed by methionine aminopeptidase (MAP) to yield mature peptides and free methionine, which is recycled. Both PDF and MAP are essential for bacterial growth, and PDF is required for MAP activity. This series of reactions is called the methionine cycle (Figure 1).

  To date, polypeptide deformylase homolog genes have been found in bacteria, chloroplast-containing plants, mice and humans. Plant proteins are encoded in the nucleus but appear to carry a chloroplast localization signal. This is consistent with the observation that the synthesis of chloroplast RNA and protein is very similar to that of eubacteria. Although there is no information on protein expression of mammalian PDF gene homologs, the functional role of such proteins has not been shown to date (Meinnel, T. 2000, Parasitology Today 16 (4), 165-168).

  Polypeptide deformylase has been found in all Eubacterium and extensive genomic sequence information is available for it. The diversity of sequences between PDF homologs is large, and there is only about 20% identity between sequences with low relevance. However, the conservation around the active site is very high, there are a few fully conserved residues, which contain one cysteine and two histidines, which cooperate with the active site metal. Necessary (Meinnel, T. et al, 1997, Journal of Molecular Biology, 267, 749-761).

  PDF is an attractive antibacterial target because it has been shown to be essential for bacterial growth in vitro (Mazel, D. et al, EMBO J. 13 (4), 914-923, 1994). However, it is not thought to be involved in eukaryotic protein synthesis (Rajagopalan et al, J. Am. Chem. Soc. 119, 12418-12419, 1997) and is conserved in prokaryotic cells. (Kozak, M. Microbiol. Rev. 47, 1-45, 1983). Therefore, PDF inhibitors may serve as broad spectrum antibacterial agents.

(Summary of Invention)
The present invention relates to novel N- [2- (N-formyl-N-hydroxy-amino) -ethyl] -arylamides represented by the following formula (I) having bacterial polypeptide deformylase inhibitory activity and PDF inhibition Relates to its use as an agent.
Furthermore, the present invention provides a method for inhibiting PDF in animals, including humans, comprising administering to a subject in need of treatment an effective amount of a compound of formula (I) below. .

［式中：
Ａは、５〜７個の原子からなり、１〜４個のヘテロ原子を含む、縮合芳香族または脂肪族環系であり、ここに、Ａは、１〜９個の炭素原子を有する置換されていてもよいアルキルまたはシクロアルキル、ハロ、１〜９個の炭素原子を有するアルコキシ、ヒドロキシ、アミノ、１〜９個の炭素原子を有するヒドロキシアルキル、アルコキシアルキル、置換されていてもよいアリールまたは置換されていてもよいヘテロアリール、カルボキシおよびアルコキシカルボニルから選択される１、２または３個の置換基により置換されていてもよく、ＸおよびＹは、独立して、ハロ、ヒドロキシまたは１〜３個の炭素原子を有するヒドロキシアルキルである］
から選択される。 (Detailed description of the invention)
Compounds useful in the methods of the present invention include compounds of the following formula (I):

[Where:
A is a fused aromatic or aliphatic ring system consisting of 5 to 7 atoms and containing 1 to 4 heteroatoms, wherein A is substituted with 1 to 9 carbon atoms Optionally substituted alkyl or cycloalkyl, halo, alkoxy having 1 to 9 carbon atoms, hydroxy, amino, hydroxyalkyl having 1 to 9 carbon atoms, alkoxyalkyl, optionally substituted aryl or substituted Optionally substituted by 1, 2 or 3 substituents selected from heteroaryl, carboxy and alkoxycarbonyl, wherein X and Y are independently halo, hydroxy or 1-3 Is a hydroxyalkyl having a carbon atom of
Selected from.

  As used herein, “alkyl” refers to optionally substituted hydrocarbon groups joined together by carbon-carbon bonds. The alkyl hydrocarbon group may be linear, branched or cyclic, saturated or unsaturated.

  As used herein, “aryl” means an optionally substituted aromatic group having a conjugated pi-electron system and containing up to two conjugated or fused ring systems. “Alkyl” includes carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which may be substituted.

Preferred compounds of the invention are:
5-chloro-4- [2- (N-formyl-N-hydroxy-amino) -ethyl] amido-benzimidazole.

  The present invention also includes pharmaceutically acceptable salts and complexes. Hydrochloride, hydrobromide and trifluoroacetate are preferred. The compounds of the present invention may contain one or more asymmetric carbon atoms and can exist in racemic and optically active forms. All these compounds and diastereomers are included within the scope of the present invention.

  The compounds and methods of the present invention will be better understood in connection with the following synthetic schemes that merely illustrate methods of making the compounds of the present invention, without limiting the scope of the invention as claimed. right.

Compounds of formula (I) can be prepared according to Scheme 1.

Aryl dicarboxylic acids (1) can be purchased or can be prepared by standard literature procedures. (1) was converted to acid chloride, and aminoacetaldehyde was aminated with dimethyl acetal to obtain amide (2). Deprotection with 6N HCl in THF and treatment of the resulting aldehyde (3) with hydroxylamine and sodium acetate in MeOH gave the oxime (4). The oxime was reduced to hydroxylamine (5) with NaCNBH ₃ in MeOH under acidic conditions. Finally, N-formyl-N-hydroxylamine (I) was obtained by treating hydroxylamine in pyridine with mixed acid anhydride formed from formic acid and acetic anhydride.
5-Chloro-4- [2- (N-formyl-N-hydroxy-amino) -ethyl] amido-benzimidazole was prepared from 5-chloro-4-carboxy-benzimidazole in a manner similar to the examples described above. did.

Preparation of 5-chloro-4-carboxy-benzimidazole a) 6-amino-2-chloro-5-nitrobenzoic acid 2,6-dichloro-3-nitrobenzoic acid (2.93 g, 12.4 mmol), copper chloride A mixture of (I) (0.025 g, 0.25 mmol) and aqueous ammonium chloride (12.5 mL) was stirred in a sealed vessel at 125 ° C. for 18 hours. The mixture was acidified with 6N HCl. The yellow solid precipitate was washed with 6N HCl to give the title compound as a mixture with 6-amino-2-chloro-3-nitrobenzoic acid. ¹ H NMR * (400 MHz, CD ₃ OD): δ 8.17 (d, j = 9.2 Hz, 1H); 6.78 (d, j = 9.2 Hz, 1H). * Main isomer.

b) Methyl 6-amino-2-chloro-5-nitrobenzoate (trimethylsilyl) diazomethane (2M in hexane, 8 mL, 16 mmol) was added to 6-amino-2-chloro- in dichloromethane (52 mL) and methanol (17 mL). To a mixture of 5-nitrobenzoic acid and 6-amino-2-chloro-3-nitrobenzoic acid (1.74 g, 8.03 mmol) was added dropwise at 0 ° C. The mixture was stirred for 30 minutes and concentrated on a rotary evaporator. The residue was purified by flash column chromatography (silica gel, 30% ethyl acetate / hexanes) to give the title compound as a yellow solid (0.85 g, 46%). ¹ H NMR (400 MHz, CD ₃ OD): δ 8.02 (d, j = 9.2 Hz, 1H); 6.61 (d, j = 9.2 Hz, 1H); 3.81 (s, 3H).

c) Methyl 2-chloro-5,6-diamino-benzoate Methyl 6-amino-2-chloro-5-nitrobenzoate (0.85 g, 3.69 mmol) and tin (II) chloride diazomethane (3.5 g, 18.4 mmol) was refluxed in methanol (25 mL) for 2 hours. The mixture was cooled to room temperature and concentrated on a rotary evaporator. The residue was partitioned between ethyl acetate and saturated aqueous potassium hydroxide. The aqueous phase was extracted with ethyl acetate (3x). The combined organic extracts were dried (MgSO ₄ ) and evaporated to give the title compound (0.73 g, 97%) as a brown oil. ¹ H NMR (400 MHz, CD ₃ OD): δ 6.54 (d, j = 8.3 Hz, 1H); 6.42 (d, j = 8.3 Hz, 1H); 3.74 (s, 3H).

d) 4-Methoxycarbonyl-5-chlorobenzimidazole Methyl 2-chloro-5,6-diamino-benzoate (0.73 g, 3.61 mmol) was refluxed in formic acid (30 mL) for 18 hours. The mixture was cooled to room temperature and concentrated on a rotary evaporator. The residue was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. The aqueous phase was extracted with ethyl acetate (3x). The combined organic extracts were dried (MgSO ₄ ). The residue was triturated from diethyl ether / hexanes to give the title compound as a tan solid (0.47 g, 60%). ESMS: M + H = 211.

e) 5-Chloro-4-carboxy-benzimidazole 4-Methoxycarbonyl-5-chlorobenzimidazole (0.46 g, 2.18 mmol) and sodium hydroxide (0.43 g, 11 mmol) in tetrahydrofuran (5 mL), methanol Refluxed in (5 mL) and water (2 mL) for 18 hours and in formic acid (30 mL) for 18 hours. The mixture was cooled and acidified with 1N HCl. The mixture was washed with ethyl acetate (2x) and evaporated under reduced pressure. The solid residue was stirred in methanol (20 mL) and filtered. The filtrate was evaporated to give the title compound (100%) as a tan solid. ESMS: M + H = 197.

The following compounds can be prepared in a similar manner as the above examples:
6-Chloro-2,3-dimethyl-N- [2- (N-formyl-N-hydroxy-amino) -ethyl] -quinoxaline-5-carboxamide 5-chloro-3-phenyl-N- [2- (N -Formyl-N-hydroxy-amino) -ethyl] -3H-benzotriazole-4-carboxamide 6-chloro-N- [2- (N-formyl-N-hydroxy-amino) -ethyl] -benzo [1,2 , 3] thiadiazole-7-carboxamide 5,6-dichloro-N- [2- (N-formyl-N-hydroxy-amino) -ethyl] -1,3-dihydro-isobenzofuran-7-carboxamide 6-chloro- 2,3-Dimethyl-N- [2- (N-formyl-N-hydroxy-amino) -ethyl] -2,3-dihydro-benzofuran-7-carboxamide.

  With the appropriate treatment and protection of any chemical functional group, the remaining compounds of formula (I) are synthesized by methods similar to those described above and in the Examples section.

  In order to use a compound of formula (I) or a pharmaceutically acceptable salt thereof in the treatment of humans and other mammals, it will normally be formulated as a pharmaceutical composition in accordance with standard pharmaceutical practice.

  The compounds of the present invention are useful for the treatment of bacterial infections, including but not limited to respiratory tract infections and / or gram positive bacterial infections.

  The compounds of formula (I) and their pharmaceutically acceptable salts are prepared in a standard way for antibiotics, eg oral, parenteral, sublingual, dermal, transdermal, rectal Can be administered by inhalation or from the buccal side.

  Compositions of the compounds of formula (I) and their pharmaceutically acceptable salts that are active when administered orally can be formulated as syrups, tablets, capsules, creams and lozenges. In general, a syrup formulation will consist of a solution of the compound or salt in a liquid carrier such as ethanol, peanut oil, olive oil, glycerin or water containing a flavor or colorant. When the composition is in tablet form, any pharmaceutical carrier commonly used in the preparation of solid formulations may be used. Examples of such carriers include magnesium stearate, white clay, talc, gelatin, acacia, stearic acid, starch, lactose and sucrose. When the composition is in capsule form, any conventional encapsulation method is suitable, for example, the carrier is used in a hard gelatin capsule shell. When the composition is in the form of a soft gelatin shell capsule, any pharmaceutical carrier commonly used for preparing dispersions or suspensions, such as aqueous gums, celluloses, silicates or oils, can be considered. include.

  A typical parenteral composition comprises a compound in a sterile aqueous or non-aqueous carrier, which may contain a parenterally acceptable oil, such as polyethylene glycol, polyvinylpyrrolidone, lecithin, peanut oil or sesame oil, or It consists of a salt solution or suspension.

  A typical inhalation composition is in the form of a solution, suspension or emulsion that can be administered as a dry powder or aerosol form using conventional propellants such as dichlorodifluoromethane or trichlorofluoromethane. is there.

  A typical suppository formulation is thus administered together with binders and / or lubricants such as polymeric glycols, gelatin, cocoa butter or other low melting vegetable waxes or oils or synthetic analogs thereof. Or a pharmaceutically acceptable salt thereof.

  Typical skin and transdermal formulations include conventional aqueous or non-aqueous vehicles such as creams, ointments, lotions or pastes, or are in the form of medicinal plasters, patches or films.

  Preferably, the composition is in unit dosage form such as a tablet, capsule or metered aerosol so that the patient can administer it at once.

  Each unit dosage for oral administration suitably contains 0.1 mg to 500 mg / kg, preferably 1 mg to 100 mg / kg, each unit dosage for parenteral administration suitably Contains 0.1 mg to 100 mg / kg of a compound of formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid. Each unit dosage for intranasal administration suitably contains 1 to 400 mg, preferably 10 to 200 mg per person. Topical formulations suitably contain 0.01-5.0% of the compound of formula (I).

The daily dosage regimen for oral administration is suitably about 0.01 mg / kg to 40 mg / kg of the compound of formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid. A daily dosage regimen for parenteral administration is suitably about 0.001 mg / kg to 40 mg / kg of a compound of formula (I) or a pharmaceutically acceptable salt thereof calculated as the free acid. . The daily dosage regimen for intranasal administration and oral inhalation is suitably about 10 to about 500 mg / person. Sufficient active ingredient may be administered 1-6 times daily to exhibit the desired activity.
When the compounds of the invention are administered according to the invention, no toxicological effects are expected.

  The biological activity of the compound of formula (I) is shown by the following test:

Biological assays:
Lazennec and Meinnel, (1997) `` Formate dehydrogenase-coupled spectrophotometric assay of peptide deformylase '' Anal. Biochem. 244, pp. 180-182. The PDF activity of E. coli is measured at 25 ° C. The reaction mixture contains 50 mM potassium phosphate buffer (pH 7.6), 15 mM NAD, 0.25 U formate dehydrogenase in 50 μl. Substrate peptide K _M concentration, include f-Met-Ala-Ser. Trigger the reaction by adding 10 nM Def1 enzyme and monitor absorbance at 340 nm for 20 minutes.

Antimicrobial activity assay:
Broth microdilution using the method recommended by the National Committee for Clinical Laboratory Standards (NCCLS), Document M7-A4, “Methods for Dilution Susceptibility Tests for Bacteria that Grow Aerobically”. The whole-cell antimicrobial activity was examined by microdilution). Compounds were tested in 2-fold serial dilutions ranging from 0.06 to 64 mcg / ml. A panel of 12 strains was evaluated in the assay. This panel consisted of the following laboratory strains: Staphylococcus aureus Oxford, Staphylococcus aureus WCUH29, Enterococcus faecalis I, Enterococcus faecalis 7, Haemophilus influenzae Q1, Haemophilus influenzae NEMC1, Moraxella catarrhalis 1502, Streptococcus pneume pneumoniae N1387, E. coli 7623 (AcrABEFD +) and E. coli 120 (AcrAB-). The minimum inhibitory concentration (MIC) was determined as the minimum concentration of compound that inhibited growth when viewed with the naked eye. A mirror reader was used to assist in determining the MIC endpoint.

  All publications, including but not limited to patents and patent applications cited herein, are hereby incorporated by reference.

The above description fully discloses the invention including preferred embodiments thereof. Modifications and improvements of the specific examples specified herein are within the scope of the claims. One skilled in the art can utilize the present invention to its full extent, using the above description, without further consideration. Accordingly, the examples herein should be construed as merely illustrative and not a limitation of the scope of the present invention in any way. Specific examples of the invention claiming exclusive ownership or privilege are set forth in the following claims.

Claims (3)

Formula (I):

[Where:
A is a fused aromatic or aliphatic ring system consisting of 5 to 7 atoms and containing 0 to 4 heteroatoms, and A is an optionally substituted 1 to 9 carbon atom Alkyl or cycloalkyl having, halo, alkoxy having 1 to 9 carbon atoms, hydroxy, amino, hydroxyalkyl and alkoxyalkyl having 1 to 9 carbon atoms, wherein the alkyl and alkylene groups are independently Having 1 to 9 carbon atoms), substituted by 1, 2 or 3 substituents selected from optionally substituted aryl or optionally substituted heteroaryl, carboxy and alkoxycarbonyl X and Y are independently halo, hydroxy or 1-3 hydroxyalkyl]
The compound shown by.   5-Chloro-4- [2- (N-formyl-N-hydroxy-amino) -ethyl] amido-benzimidazole. A method of treating a bacterial infection by administering to a subject the compound of claim 1.