KR101828665B1 - A derivative of benzylidenehydrazine and a use of the derivative as inhibitor of beta-Ketoacyl-acyl carrier protein synthase III derived from Gram-negative bacteria - Google Patents

Abstract

The present invention relates to the use of a novel benzylidenehydrazine derivative having an inhibitory activity of β-ketoacyl-acyl carrier protein synthase III (KAS III).

Description

Benzylidene hydrazine derivative and its use as an inhibitor of beta-ketoacyl-acyl group transfer protein-producing enzyme III derived from Gram-negative bacteria {A derivative of benzylidenehydrazine and a derivative thereof as a derivative of beta-ketoacyl-acyl carrier protein synthase III Gram-negative bacteria}

The present invention relates to a benzylidenehydrazine derivative (hereinafter referred to as " KAS III ") having an inhibitory activity against beta-ketoacyl-acyl carrier protein synthase III .

Since penicillin, a number of antibiotics have been developed and used to combat bacterial infiltration into the body. Recently, however, strains resistant to these antibiotics have emerged and are regarded as a big problem. Bacterial species such as Enterococcus faecalis , Mycobacterium tuberculosis and Pseudomonas aeruginosa , which can pose a life threat, are resistant to all known antibiotics (Stuart B. Levy, Scientific American , 46-53, 1998).

Conventional chemical antibiotics, such as penicillin, cephalosporin, etc., exhibit antibiotic action by inhibition of cell wall or protein synthesis of microorganisms. It is urgent to develop an antibiotic having a new action mechanism capable of exterminating these resistant strains.

Recently, P. aeruginosa and A. baumannii, which are the most common gram-negative bacteria, are known to be resistant to antibiotics due to endogenous resistance and most frequently occurring bacteria. In P. aeruginosa and A. baumannii , multidrug resistance means a strain resistant to all three classes of drugs such as aminoglycoside, fluoroquinolone and carbapenem. Cabapenem was the only effective antibiotic, The increase in the number of strains resistant to phenem appears to have a major limitation in antibiotic treatment. Pseudomonas or Pseudomonas aeruginosa aeruginosa is a gram-negative bacterium that is a common infectious pathogen and has low susceptibility to antibiotics. Therefore, it can easily acquire antibiotic resistance naturally. It can be used for respiratory tract infection, burn infection, cystic fibrosis pneumonia, urinary tract infection And it is present in the infected cattle such as artificial ring apparatus, suction catheter, and contaminated water, and it is directly or indirectly propagated to the patient. In the case of patients with advanced disease, treatment is difficult, .

In the chemotherapy of bacterial infections, antibiotics such as rifampicin, kanamycin, streptomycin, biomycin, capreomycin and cycloserine have been used as antibiotics. In chemotherapy of bacterial infectious diseases, it is a serious problem that the bacteria causing the infectious diseases become drug resistant. There is a strong desire for new chemotherapeutic agents that are effective against the infections of drug resistant resistant acid bacteria. In addition, new chemotherapy regimens effective against infectious diseases of atypical antibiotics, which have not been established for chemotherapy, are strongly desired. Thus, unlike known antibiotics conventionally used, It is strongly desired to find or manufacture a novel compound which exhibits good properties such as action.

In general, the beta-ketoacyl-acyl transfer protein-producing enzyme III is a protein that initiates the synthesis of fatty acid of bacteria. It is a protein that receives an acyl group from a malonyl-ACP and converts it into a beta-ketoacyl- (β-Ketoacyl-ACP) III (FIG. 1).

The formation of the beta-ketoacyl-acyl transferase III protein triggers fatty acid synthesis, which results in the formation of bacterial or mammalian cell walls. Therefore, beta-keto-acyl-acyl transfer protein produced through the inhibition of the enzyme III shows the antimicrobial effect by inhibiting the cell wall formed of several bacteria (Prog Lipid Res 2001, 40, 467; J. of Biol Chem 2003,.... 19, 51494).

In 2001, Professor Charles O. Rock of the University of Tennesse, USA, reported that thiolactomycin, an inhibitor of type II fatty acid synthesis, exhibited antibiotic activity through the inhibition of KAS III protein, suggesting that KAS III is an important target protein . This content was published in Journal of Biological Chemistry ( J. of Biol . Chem . 2001, 276 , 6551-6559).

In WO / 2002/055661, a beta-ketoacyl-acyl group transfer protein-producing enzyme III inhibitor was developed by mimicking a beta-lactam-based compound used as a conventional antibiotic and found to be useful as an antibiotic. Recently, in WO / 2008/061399, oxo-dihydro pyrrol carboxylic acid and derivatives thereof have been proposed as novel beta-ketoacyl-acyl group transfer protein-forming enzyme III inhibitors and their antibiotic activity has been demonstrated. Also, in 2002, Smith Kline, a multinational pharmaceutical company, found that dichloro phenyl-acetic acid-based compounds inhibit the beta -ketoacyl-acyl transfer protein transferase III in WO / 2002/09651 and are useful as excellent antibiotics. In WO 2004/041189 in 2004, furancarboxylic acid and its derivatives inhibit beta-ketoacyl-acyl group-forming enzyme III to inhibit fatty acid production and thus exert an excellent effect on breast cancer and prostate cancer, thus being useful as anticancer agents for these inhibitors . In the 2009 10-2009-0031869 patent, it has been demonstrated that floretin, a kind of flavonol, is easy to utilize as an antibiotic by inhibiting beta-ketoacyl-acyl transfer protein transferase III. 2014 Eur . J. Med . Chem . 76, 387-396, found that 2-Styryl-5-Nitroimidazole derivatives have an excellent inhibitory effect on beta-ketoacyl-acyl group transfer protein-producing enzyme III.

Recently, Hai-Liang Zhu professor team of Nanjing University has been working on the discovery of beta-ketoacyl-acyl transfer protein-forming enzyme III inhibitor, and it is also interested in discovering efficacy substances for Gram-negative bacteria. In these cases, the finding of various skeletal structures that have an effect on E. coli and P. aeruginosa among Gram-negative bacteria has been consistently reported in the literature ( Eur . J. Med . Chem . , 2014, 76, 387-396 ; Bioorg . Med . Chem . , 2013, 23, 4235-4238).

Since 2005, the present inventors have been steadily developing antibiotics through inhibition of beta-ketoacyl-acyl group-forming enzyme III, and have succeeded in discovering substances having broad antibiotic activity in various bacteria. In 2009, YKAs3003 containing a dihydroxy-benzyl structure was discovered and registered as a patent (10-1145206) and a paper was published ( Bioorg . Med . Chem. , 2009, 17, 1506-1513). Flavonoids and natural products based antibiotics Phloretin (YKAF04) was discovered (Patent Registration No. 10-1149322) through the progress of the development research, and the content was published in the SCI paper ( Bioorg. Med . Chem . , 2009, 17, 5408-5413). We have succeeded in finding YKA3028, a substance with excellent antibiotic effect on Gram-positive S. aureus and methicillin-resistant Staphylococcus aureus (MRSA), patent registration (10-1145612) and Eur . J. Med . Chem . , 2012, 47, 261-269, and YKA3028 was used as the parent material of this patent.

In the case of YKA3028, the previous patent has proposed efficacy limited to Gram-positive bacteria, but this patent has confirmed that it has novel effects on Gram-negative bacteria.

[Prior Patent Literature]

Korean Patent Publication No. 10-2015-0132805

Korean Patent Publication No. 10-2013-0087118

The present invention has been made in view of the above needs, and an object of the present invention is to provide a compound which inhibits Gram-negative bacteria-derived beta-ketoacyl-acyl group transfer protein-producing enzyme III.

Another object of the present invention is to provide a compound that can be used as a gram negative multifunctional anti-microbial target antibiotic by inhibiting the fatty acid synthesis of Gram-negative bacteria.

To achieve the above object, there is provided a benzylidenehydrazine derivative selected from the group consisting of the following chemical formulas (1), (2) and (3).

[Chemical Formula 1]

(2)

(3)

The present invention also provides a pharmaceutical composition comprising the above-mentioned compound, a derivative thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

In the present invention, 'derivative' is the most used compound, and is a similar compound obtained by chemically changing a part of a compound. Usually refers to a compound in which a hydrogen atom or a specific atomic group in a compound is substituted by another atom or atomic group.

In one embodiment of the present invention, the composition preferably inhibits beta-ketoacyl-acyl group transfer protein-producing enzyme III (KAS III), but is not limited thereto.

The present invention also provides a pharmaceutical composition for preventing or treating an infectious disease caused by a microorganism comprising an effective amount of the compound of Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

In one embodiment of the present invention, the microorganism is preferably Gram negative bacteria, and the Gram negative organism is Pseudomonas aeruginosa Aeruginosa), Acinetobacter baumannii (Acinetobacter baumannii ) from But is not limited thereto.

The antibiotic-resistant strain may be a strain resistant to at least one antibiotic selected from the group consisting of penicillin antibiotics, methicillin antibiotics, quinolone antibiotics, vancomycin antibiotics, carbapenem antibiotics, and aminoglycoside antibiotics Specifically, methicillin-resistant Staphylococcus aureus (MRSA), quinolone-resistant Staphylococcus aureus (QRSA), vancomycin resistant enterococcus resistant enterococcus (VRE), vancomycin-resistant S. aureus (VISA) or Acinetobacter baumannii, a multidrug-resistant strain.

Unless otherwise stated, the term "prophylactic" or "treatment ", as used herein, refers to reversing, alleviating, or progressing the disease or condition to which the term applies, or one or more symptoms of the disease or condition ≪ / RTI >

Unless otherwise indicated, the term " bacterial infection ", or "infection" or "infectious disease ", as used herein,

Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecalis, Enterococcus faecalis, E. faecium, E. casselflavus, S. epidermidis, S. hemolyticus, or Peptostreptococcus species, including E. faecium, E. faecium, E. casshellavus, S. epidermidis, Staphylococcus hemolyticus, Streptococcus pyogenes, C and G streptococci, Corynebacterium spp., Pneumonia, Sinusitis, Bronchitis, Tonsillitis and mastoiditis associated with infection by spp. It has been reported that infection with Corynebacterium diptheriae or Actinobacillus haemolyticum Associated with infection by Mycoplasma pneumoniae, Legionella pneumophila, Streptococcus pneumoniae, Haemophilus influenzae or Chlamydia pneumoniae, including, but not limited to, rhinitis, rheumatic fever and glomerulonephritis Including strains resistant to known antibiotics such as but not limited to respiratory tract infections; beta-lactam, vancomycin, aminoglycosides, quinolones, chloramphenicol, tetracycline and markrolide. Blood and tissue infections, including endocarditis and osteomyelitis, caused by E. faecalis, E. coli, E. coli, E. coli, E. coli, E. coli, E. coli, Aureus, coagulase-negative staphylococci (i.e., staphylococci Epidermidis, Staphylococcus H. ET Mora coarse, etc.), Streptococcus Pio jeneseu, Streptococcus Agar lock thiazole (Sterptococcus agalactiae), Streptococcus group C-F (Streptococcal groups C-F; Coli Streptococcus), viridans streptococci, Corynebacterium minutissimum, Clostridium spp., Or Bartonella henselae, and the like. , Simple skin and soft tissue infections and abscesses associated with infection by an infectious agent such as Staphylococcus aureus, coagulase-negative staphylococcus species, or enterococcus spp. A simple acute urinary tract infection associated with infection, such as urethritis and cervicitis; Chlamydia trachomatis, Haemophilus ducreyi, Treponema pallidum, Ureaplasma urealyticum ) Or Neisseria gonorrheae; a disease which is transmitted to a sexual activity related to an infection by Neiserria gonorrheae; a disease of Staphylococcus aureus Food poisoning and toxin shock syndrome) or toxin diseases associated with infection by streptococcal strains A, B and C; ulcers associated with infection by Helicobacter pylori; infection by Borrelia recurrentis; Related systemic fever syndrome; Lyme disease associated with infection by Borrelia burgdorferi; Chlamydia trachomatis, Naceria gnorhea, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus Mycobacterium avium or Mycobacterium intracellulare, which is associated with infections caused by P. aeruginosa, P. aeruginosa, P. aeruginosa, P. aeruginosa, P. aeruginosa, P. aeruginosa, P. aeruginosa, P. aeruginosa, P. aeruginosa, P. aeruginosa, Infectious mycobacterial iron complex (MAC) disease associated with infection; Mycobacterium tuberculum System (Mycobacterium tuberculosis), Mycobacterium Le Prado (M. leprae, M. paratuberculosis, M. kansasii, or M. chelonei; or an infectious disease caused by M. tuberculosis; Gastroenteritis associated with infection by Campylobacter jejuni; intestinal protozoa associated with infection by Cryptosporidium spp.; Bacterial infection associated with infection by viridans streptococcus; Bordetella pertussis < RTI ID = 0.0 > Chronic cough associated with infection by Bordetella pertussis; gas circulation associated with infection by Clostridium perfringens or Bacteroides spp .; And atherosclerosis or cardiovascular disease associated with infection by Helicobacter pylori or Chlamydia pneumoniae. Bacterial and protozoal infections that can be treated or prevented in an animal, and diseases associated with the infection include, Diseases include: respiratory diseases of cattle associated with infection by Pasteurella haemolytica, P. multocida, Mycoplasma bovis or Bordetella spp; Cryptococcal aureus, Strep. Uberis, Streptococcus agalactia, Streptococcus (Streptococcus agalactiae), Streptococcus agalactiae, Streptococcus agalactiae, Streptococcus agalactiae, Streptococcus agalactiae, Mastitis of a cow associated with infection by Streptococcus sp .; Strep. Dysgalactiae, Corynebacterium or Enterococcus; Swine respiratory diseases associated with infection by Actinobacillus pleuropneumoniae, Pasteurella mutosidia or Mycoplasma spp .; Lawsonia intracellularis, Salmonella or Serpulina Pseudomonas infection associated with infection by Fusobacterium spp.; Fusobacterium necrophorum or Bacteroides nodosus; Fusobacterium < RTI ID = 0.0 > necrophorum < / RTI & Hairy warts associated with infection by bovine spongiform encephalopathy associated with infection by bovis (Moraxella bovis), preterm premature cow associated with infection by protozoan (ie, neosporium), staphylococcus Epidermidis, staphylococcus intermedia, coagulase enzyme negative staphylococcus sp. Par is far stew Relais skin and soft tissue infections in dogs and cats Toshi is related to infection by; And by Alcaligenes spp., Bacteroides species, Clostridium species, Enterobacter species, Eubacterium, Peptostreptococus, Porphyromanas or Prevotella. Tooth or mouth infection in dogs and cats associated with infection.

Other bacterial infections and protozoal infections that may be treated or prevented according to the methods of the present invention and diseases associated with such infections are described in JP Sanford et al., "The Sanford Guide To Antimicrobial Therapy ", 26th Ed., (Antimicrobial Therapy , Inc. (1996).

In one embodiment of the present invention, the infectious disease is preferably selected from the group consisting of staphylococcus food poisoning, bonglesulitis, lymphatic tubular infections, urinary tract infections, meningitis, peritonitis, cystitis, respiratory diseases, otitis media, pneumonia, pyogenic inflammation and sepsis But not limited to this.

In one embodiment of the present invention, the effective amount is preferably 2 to 512 μg / mL, but is not limited thereto.

The compound of the formula of the present invention can be prepared from a production method known in the art by using a starting material and a catalyst known in the art and can be directly extracted from soybean plants such as 칡, licorice and the like. In one embodiment of the present invention, a virtual screening was performed using a computer in a database as described in Example 1 and then purchased from a vendor (ChemBridge Corp., San Diego, USA).

Hereinafter, the present invention will be described.

The present invention relates to a novel compound which inhibits beta-ketoacyl-acyl group transfer protein-producing enzyme III and uses of the compound, and exhibits an antibiotic effect by inhibiting bacterial cell wall formation and destroying bacteria.

Hereinafter, the present invention will be described in detail.

In one embodiment of the present invention, when optically active centers are present in the compounds of the present invention, the present invention provides, as individual specific embodiments, all individual optically active forms and their mixed forms, and their corresponding racemates, do. Racemates may be prepared by known methods including the formation of diastereomeric derivatives with suitable optically active auxiliary groups and subsequent separation and resolution of auxiliary groups (see Advanced Organic Chemistry: 3rd Edition: author J March, p. 104-107) And can be separated into individual optically active forms.

The compounds according to the invention may contain one or more asymmetrically substituted carbon atoms. The presence of such one or more asymmetric centers (chiral centers) in the compounds of the above formula may lead to stereoisomers, and in each case the present invention encompasses all stereoisomers, including enantiomers and diastereomers, And mixtures thereof.

When tautomers (tautomers) are present in the compounds of the present invention, the present invention discloses all individual tautomeric forms and their mixed forms as individual specific embodiments.

As described above, the compound of the present invention is a beta-ketoacyl-acyl group precursor protein-producing enzyme III inhibitor.

The compounds of the present invention may be provided as pharmaceutically acceptable salts. As these salts, acid addition salts formed by pharmaceutically acceptable free acids are useful. The compound represented by the above formula may form a pharmaceutically acceptable acid addition salt according to a method conventional in the art. As the free acid, organic acids and inorganic acids can be used. As the inorganic acids, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid and the like can be used. As the organic acids, citric acid, acetic acid, lactic acid, tartaric acid, fumaric acid, formic acid, propionic acid, oxalic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid or aspartic acid Can be used.

In another aspect, suitable salts are alkali metal salts, such as sodium or potassium, alkaline earth metal salts such as calcium or magnesium, or organic amine salts, such as base salts such as triethylamine.

They may also be provided as in vivo hydrolysable esters. These are pharmaceutically acceptable esters that hydrolyse in the body to produce parent compounds. Such esters can be identified, for example, by intravenously administering a test compound to a test animal and then examining the body fluid of the test animal. Suitable in vivo hydrolysable esters for carboxy include methoxymethyl and those for hydroxy include formyl and acetyl, especially acetyl.

Use of the beta-ketoacyl-acyl group transfer protein-producing enzyme III inhibitor compound of the present invention or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof for the therapeutic treatment (including prophylactic treatment) of mammals including humans , It is generally formulated as a pharmaceutical composition according to standard pharmaceutical guidelines.

Thus, in another aspect, the invention provides a pharmaceutical composition comprising a compound of the invention, or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, and a pharmaceutically acceptable carrier.

The product according to the invention can be used as a medicament, for example in the form of a pharmaceutical preparation for enteral or parenteral administration. For example, the product according to the invention may be orally administered in the form of tablets, coated tablets, dragees, hard and soft gelatine capsules, solutions, emulsions or suspensions; To work in the same form as a suppository; Or may be administered parenterally, such as in the form of injection solutions.

The pharmaceutical preparations can be prepared by combining the substances according to the invention with other substances useful for the treatment of the disease, in combination with suitable non-toxic, inert, therapeutically synthetic solid or liquid carrier materials and, if desired, conventional pharmaceutical additives, , Which may be produced in a manner familiar to one of ordinary skill in the art.

Both inorganic and organic carrier materials are suitable as such carrier materials. Thus, for example, lactose, corn starch or derivatives thereof, talc, stearic acid or its salts can be used as carrier materials for tablets, coated tablets, dragees and hard gelatine capsules. Examples of suitable carriers for soft gelatine capsules are vegetable oils, waxes, fats and semi-solid and liquid polyols (however, depending on the nature of the active ingredient no carrier is required in the case of soft gelatine capsules). Examples of carrier materials suitable for the production of solutions and syrups include water, polyols, sucrose, phosgene and glucose. Examples of suitable carrier materials for injection solutions are water, alcohols, polyols, glycerol and vegetable oils. Examples of suitable carrier materials for suppositories are natural or hardened oils, waxes, fats and semi-solid or liquid polyols.

Conventional preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavors, salts for varying the osmotic pressure, buffering agents, masking agents and antioxidants are contemplated as pharmaceutical adjuncts.

For oral administration, the compounds of the above formula and their respective salts are preferably provided as a lyophilizate or anhydrous powder with a conventional carrier such as water and isotonic saline for dilution.

The pharmaceutical compositions of the present invention may be administered by standard methods for the disease or condition to be treated, for example, by oral, topical, parenteral, ball, nasal, vaginal or rectal administration or by inhalation. To this end, the compounds of the present invention may be formulated for administration by any means known in the art, for example, as tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, gels, intranasal sprays, suppositories, , And sterile aqueous or oily solutions or suspensions or sterile emulsions for parenteral use (including intravenous, intramuscular, or infusion).

The pharmaceutical compositions of the present invention may contain, or may be co-administered (concurrently or sequentially), one or more pharmaceutical agents useful for treating one or more of the above-mentioned one or more diseases or conditions, in addition to the compounds of the present invention.

The pharmaceutical compositions of the present invention will generally be administered to humans in a daily dose ranging from 0.1 to 100 mg / kg body weight (preferably 0.5 to 50 mg / kg body weight). Such daily doses may be administered in divided doses as necessary, and the exact amount and route of administration of the compound administered will depend upon the body weight, age and sex of the patient being treated and the particular disease or condition being treated, in accordance with principles well known in the art. do.

Unit dosage forms will typically contain from about 1 mg to 1000 mg of a compound of the invention. Therefore, in another aspect, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, for use in a method of therapeutic treatment of humans or animals or as a therapeutic agent. The present invention discloses use in the treatment of a disease or condition mediated by one or more beta-ketoacyl-acyl transfer protein-producing enzyme III.

It is contemplated that the invention described herein is not limited to the particular methods, protocols, and reagents described as being capable of being varied. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the invention in any way.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices, and materials are described below.

In the practice of the present invention, a number of common techniques of molecular biology and cell biology are used. These techniques are well known and are described, for example, in Current Protocols in Molecular Biology, Volumes I, II and III, 1997 (F. M. Ausubel ed.); Sambrook et al., 1989, Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y.).

As can be seen from the above-mentioned solution and the following examples, the present invention ultimately utilizes a compound inhibiting beta-ketoacyl-acyl group transfer protein-producing enzyme III as an antibiotic by inhibiting fatty acid synthesis.

Brief Description of the Drawings Fig. 1 shows the reaction of beta-ketoacyl-acyl group transfer protein production by KAS III protein in bacteria,
FIG. 2 is a graph showing the results of eight kinds of benzylidene hydrazine derivatives selected through structure-activity correlation analysis,
Figure 3 shows A. baumannii A graph of the binding constant measurement result of YKab-6 for KASIII,
Figure 4 shows the results of antimicrobial activity through measurement of minimal inhibitory concentration (MIC) in various bacteria,

The present invention will now be described in more detail by way of non-limiting examples. The following examples are provided to aid understanding of the present invention, and the scope of the present invention is not construed as being limited by the following examples.

Example  One: Virtual Drug Discovery  One of the technologies - structure discovery through active correlation analysis

YKA3028 (4 - [(5-Trifluoromethyl-pyridin-2-yl) -hydrazonomethyl] -benzene-1,3-diol (Patent Registration No. 10-1145612) Structure-Activity Relationship (SAR) analysis was performed based on benzylidenehydrazine, which is the parent structure of the drug, and the active skeletal map determined in the search for a new drug. Gram-negative A. baumannii The possible beta-ketoacyl-acyl transfer protein-producing enzyme III inhibitor was re-selected.

Benzylidenehydrazine A chemical structural change in the parent structure was designated as R and a total of five chemical transformations were induced. In the case of Rl to R5, the active skeleton map previously determined has a spatial margin to enable interaction with the protein or modification of the substituent. A variety of modifications to the position of each R can suggest structural and chemical factors that play a crucial role in demonstrating the KAS III protein-induced Gram-negative bacteria-induced inhibition. The degree of substitution at each R position is tabulated in Fig.

The library for virtual drug discovery was configured in the form of Catalyst / CatDB from Accelrys, and the compounds were able to find multiple conformation of the three dimensional structure and control the parameters to include all chemically possible conformers.

Using the Cerius2 / SBF module of Accelrys of the present invention, seven kinds of candidate substances (YKab-1 to YKab-7) were finally selected as Gram-negative bacteria KAS III inhibitory substance using a single map.

Example 2: Calculation of binding constants using fluorescence spectroscopy

10 μM of Gram-negative bacteria, E. coli KAS III and A. baumannii KAS III and P. aeruginosa KAS III were dissolved in 150 mM sodium phosphate, 100 mM NaCl, pH 8.0 buffer, and concentration and fluorescence intensity were recorded while adding the inhibitor in small amounts until the final concentration of protein and inhibitor was 1: 100 Respectively. The coupling constant K _d was calculated using the following equation.

Where F ₀ and F are the intensities of the fluorescence signals detected at 324 nm with and without inhibitor, respectively, and n is the number of inhibitors in the protein.

In the case of YKab-4 and YKab-6 among the seven candidates for KAS III, three kinds of Gram-negative bacteria, E. coli KAS III and A. baumannii KAS III, P. aeruginosa The binding constant (K _d ) for KAS III was found to be a strong binding agent to KAS III by exhibiting a binding constant of 10 ^-6 to 10 ^-9 M. It was also confirmed that YKab-0 (YKA3028), a parent compound, exhibited binding constants similar to those of the two compounds. The experimental results are shown in Table 1 and FIG.

Compound code Binding affinity test K _d E. coli KAS III A. baumannii KAS III P. aeruginosa KAS III YKab-1 6.26 × 10 ^-7 M 3.56 × 10 ^-6 M 1.07 × 10 ^-6 M YKab-2 2.56 x 10 < ^-8 > M 2.74 × 10 ^-7 M 2.30 × 10 ^-7 M YKab-3 9.89 × 10 ^-6 M 1.90 × 10 ^-6 M 2.06 × 10 ^-5 M YKab-4 6.40 × 10 ^-8 M 6.41 × 10 ^-7 M 8.54 × 10 ^-7 M YKab-5 1.78 × 10 ^-6 M 4.72 × 10 ^-6 M 4.63 × 10 ^-6 M YKab-6 2.95 × 10 ^-8 M 6.76 × 10 ^-9 M 1.58 x 10 < ^-7 > M YKab-7 4.81 × 10 ^-7 M 2.23 × 10 ^-6 M 1.30 × 10 ^-7 M YKab-0 (YKA3028) 2.83 × 10 ^-8 M 3.96 × 10 ^-8 M 2.60 × 10 ^-7 M

Table 1 shows that E. coli KAS III, A. baumannii KAS III, P. aeruginosa Binding affinity for KAS III

Example  3: minimal inhibitory concentration, MIC ) To identify antibiotic activity

It is possible to measure the degree of growth of microorganisms in a control plate and an antibiotic-added plate that do not contain antibiotics under the condition of homogeneous bacterial concentration. The results are summarized through at least three repeated experiments, (micro-dilution method).

The antibiotic effect was first measured for the seven selected compounds and the parent compound YKab-0. The bacteria used in the experiment were Gram-negative bacteria, Escherichia coli , Acinetobacter baumannii, Pseudomonas aeruginosa to, and the multi-drug resistant bacteria by MDRST 8009 (R), MDREC 1229 (R), MDRPA 2002 (R), MDRAB12035 (R) to the object.

As a result of the antibiotic activity measurement, YKab-6 showed activity of less than 32 μg / mL in three kinds of bacteria used for the activity measurement. Especially, A. baumannii showed excellent effect of 8 μg / mL in Gram-negative bacteria and E. coli And P. aeruginosa showed efficacy of 32 μg / mL. In the case of YKab-4, the gram-negative bacteria, A. baumannii 2 ug / mL efficacy and 4 ug / mL efficacy in P. aeruginosa . In addition, YKab-0 (YKA3028), a parent compound, showed excellent efficacy of 4 μg / mL in P. aeruginosa and 16 μg / mL in A. baumannii , another gram-negative organism. The experimental results are shown in Table 2.

Compound code Minimum inhibitory concentration MIC (μg / mL) E. coli A. baumannii P. aeruginosa YKab-1 > 32 > 32 > 32 YKab-2 > 32 > 32 > 32 YKab-3 > 32 > 32 > 32 YKab-4 > 32 2 4 YKab-5 > 32 > 32 > 32 YKab-6 32 8 32 YKab-7 > 32 > 32 > 32 YKab-0
(YKA3028)
> 32 16 4

Table 2 shows the antibiotic activity of the compounds against gram negative bacteria

Antimicrobial activity against these resistant bacteria was measured for these 8 substances. All three compounds that showed antimicrobial activity against Gram - negative bacteria showed antimicrobial activity of 2 ~ 16μg / mL against various strains of A. baumannii - derived resistant bacteria. In particular, in the case of YKab-4, P. aeruginosa It was confirmed that the antibiotic activity of 32 μg / mL was also exhibited for the two types of resistant multidrug-resistant bacteria.

Minimum inhibitory concentration MIC (μg / mL) MDRST 8009 (R) MDREC 1229 (R) MDRPA 2095 (R) MDRPA 2163 (R) MDRAB
12035 (R) MDRAB
12037 (R) YKab-1 > 32 > 32 > 32 > 32 > 32 > 32 YKab-2 > 32 > 32 > 32 > 32 > 32 > 32 YKab-3 > 32 > 32 > 32 > 32 > 32 > 32 YKab-4 > 32 > 32 32 32 2 2 YKab-5 > 32 > 32 > 32 > 32 > 32 > 32 YKab-6 > 32 > 32 > 32 > 32 16 16 YKab-7 > 32 > 32 > 32 > 32 > 32 > 32 YKab-0
(YKA3028) > 32 > 32 > 32 > 32 16 16

Table 3 shows the antibiotic activity of compounds against antibiotic Gram-negative resistant bacteria

Claims (13)

A benzylidenehydrazine derivative represented by the following formula (1).

[Chemical Formula 1] delete A pharmaceutical composition for preventing or treating an infectious disease caused by a microorganism comprising an effective amount of a compound represented by the following formula (1) or (2), or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

(2) The pharmaceutical composition according to claim 3, wherein the pharmaceutical composition inhibits beta-ketoacyl-acyl group transfer protein-producing enzyme III (KAS III). The pharmaceutical composition according to claim 3, wherein the microorganism is gram-negative organism. The pharmaceutical composition for preventing or treating infectious diseases caused by microorganisms according to claim 3 or 5, wherein the microorganism is a microorganism selected from the group consisting of E. coli, P. aeruginosa, A. baumannii, . 4. The method of claim 3, wherein the infectious disease is selected from the group consisting of Staphylococcus aureus food poisoning, Bongsulitis, A pharmaceutical composition for therapeutic use. The pharmaceutical composition according to claim 3, wherein the effective amount is 2 to 32 μg / mL. delete delete delete delete delete

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