WO2013042140A2

WO2013042140A2 - Non antibiotic,non peptide compounds for antibiotic efficacy & safety enhancement

Info

Publication number: WO2013042140A2
Application number: PCT/IN2012/000634
Authority: WO
Inventors: Manu Chaudhary
Original assignee: Manu Chaudhary
Priority date: 2011-09-23
Filing date: 2012-09-24
Publication date: 2013-03-28
Also published as: WO2013042140A4; WO2013042140A3

Abstract

The present invention provides one or more non antibiotic non peptidic entity to augment antimicrobial activity of one or more antimicrobial agent for combating microbial resistance The non antibiotic non peptidic entity is used individually or in combination with one or more antibiotic as single unit dose or is presented through a suitable medium The non antibiotic non peptidic entity make one or more accompanying antibiotic effective by rendering eradication of antimicrobial resistance broadening spectrum and reducing multi bacterial resistance

Description

NON-ANTIBIOTIC, NON PEPTIDE COMPOUNDS FOR ANTIBIOTIC EFFICACY & SAFETY ENHANCMENT

FIELD OF INVENTION

[0001] The embodiments herein generally relates to non-antibiotic, non-peptidic compound when used in combination with any antibiotic components enhance the activity of antibiotic compounds. More specifically the embodiments relates to different natural or synthetic compounds that can be used for reducing phenomenon of antibacterial resistance, antibiotic related side effects and drugs induced toxicities. The synergistic activity of one or more non- antibiotic, non-peptidic compounds with one or more antibiotic(s), when administered together as one product or as solvent systems is useful for the treatment of multi drug resistant microbial infections with enhanced efficacy and safety.

BACKGROUND OF THE INVENTION

[0002] Antibiotic resistance is an ever-growing clinical problem of the modern time. Several studies found that antibiotics, under the current medical practices, are over prescribed for infectious disease conditions. Compounding the issue is the fact that bacteria are learning to tolerate and even circumvent existing classes of antibiotics. Thus, multi-drug resistant bacteria remain an under recognized epidemic. In light of increasing antibiotic resistance in bacteria and the emergence of multidrug resistant pathogens such as MRSA, Pseudomonas aeruginosa, A baumanii, E coli, pneumoniae and the like need for new antibiotics is obvious. However, with reduced interest from pharmaceutical companies and the decade long process of securing drug regulatory approval, the antibiotic pipeline has run dry.

[0003] In general, the different classes of antibiotics affect five major targets: the bacterial cell wall, the cell membrane, protein synthesis, DNA and RNA synthesis, and folic acid metabolism. For example, penicillins, cephalosporins and carbapenems are classified as B-lactam antibiotics, which kill bacteria by disrupting synthesis of the bacterial cell wall. Resistance to antibiotics is a natural bacterial phenomenon that results from the evolutionary selective pressure that comes hand in hand with continued exposure to such compounds. Bacteria achieve antibiotic resistance through four general mechanisms', target modification, efflux, immunity such as biofilm formation, bypass such as conjugation, enzyme-catalyzed destruction. Hence, the use of antibiotics paradoxically accelerates its disuse. For instance, P. aeruginosa possess numerous families of efflux pumps with over- lapping substrate ranges that effectively pump out antibiotics. In total, with insensitivity to almost all commercially available antibiotics, P. aeruginosa is a particularly worrisome opportunistic pathogen. The rationale for the use of antibiotic combinations is based largely on empirical success in therapy, particularly in preventing the emergence of antibiotic resistance.

[0004] Combinations or cocktails of antibiotics are often used to broaden the antimicrobial spectrum of each and to achieve synergistic effects as disclosed in Indian Patent no. 236996, 235775, Indian Patent Application no. 308/DEL/2005 and 21 84/DEL/2008 of the same inventor. This approach has successfully been applied to combat several infectious diseases, and to prevent resistance development. But still this approach has not been a one-stop solution to overcome the problem of resistance built- up by the bacteria towards antibiotics.

[0005] Hence, there is an immense need to identify fresh approach to provide a one- stop solution to effectively address growing antimicrobial resistance and related issues of use and efficacy of antibiotics.

SUMMARY OF THE INVENTION

[0006] In view of the foregoing, a non-antibiotic, non-peptidic entity to potentiate activity of one or more antibiotic(s) in treatment of infection caused by one or more microbial resistant strain is provided. The entity is selected from the group comprising an effective amount of EDTA disodium, sodium tetraborate, Sodium salicylate, boric acid or a pharmaceutically acceptable salt thereof.

[0007] The non-antibiotic, non-peptidic entity entity may be present in range of about 0. 15 % to about 15% of said antibiotic.

[0008] Each of the non-antibiotic, non-peptidic entity may be present in range of about 0.0015 to 1 .5% in according to another preferred embodiment herein.

[0009] The amount of the non-antibiotic, non-peptidic entity may not exceed l Omg/ml or 1 to 1.5 % of total of a formulation wherein the non-antibiotic, non-peptidic entity is

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used to augment antibiotic activity of one ore more antibiotic component.

[00010] The microbial resistant strain may be selected from the group comprising Extended-spectrum beta- lactamase (ESBL), Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Enterococcus faecium(VRSA), Vancomycin- intermediate Staphylococcus aureus (VISA), Vancomycin-resistant Enterococci (VRE), New Delhi metallo-beta-lactamase 1(NDM-1), metallo-beta-lactamase producing bacteria (MBL) strain, Glycopeptide-intermediate Staphylococcus aureus (G1SA), aminoglycoside resistance strains, efflux over expressed strains, bio-film producing strains etc.

[00011] he non-antibiotic, non-peptidic entity may further comprises citric acid, tannic acid, reserpine, arylpiperazines, alkylaminoquinolones, nocardamines or a pharmaceutically acceptable salt thereof.

[00012] The non-antibiotic, non-peptidic entity may be administered either together as a single unit dose with said antibiotic or is administered separately before/after/during administration of said anti-biotic according to an preferred embodiment herein.

[00013] The non-antibiotic, non-peptidic entity may be delivered in route selected from the group comprising topical, oral, parenteral and ocular route.

[00014] In a further embodiment of the invention, the non-antibiotic, non-pept idic entity may be presented in a form selected from the group comprising nano-particles, dry powder, gel, solution or a pharmaceutically acceptable forms thereof.

[00015] In another aspect of the invention, a synergistic antibiotic composition for treatment and containment of bacterial resistance is provided. The composition may include a non-antibiotic, non-peptidic entity and one or more antibiotic component wherein proportion of said non-antibiotic, non-peptidic entity and said antibiotic component is in a range of about 0.15 % to about 15% of said antibiotic. The non- antibiotic, non-peptidic entity may be selected from the group comprising an effective amount of EDTA disodium, sodium tetraborate, Sodium salicylate, boric acid or a pharmaceutically acceptable salt thereof. The antibiotic - component may be selected form the group comprising penicillins, cephalosporins, carbapenems, aminoglycosides, sulfonamides, quinolones, tetracyclines, macrolides, glycopeptide , oxazolidinones or combinations thereof.

[00016] The non-antibiotic, non-peptidic entity may be present either as a single unit dose with said antibiotic component or is present as a suitable solvent medium according to the above mentioned embodiment.

[00017] The non-antibiotic, non-peptidic entity as mentioned above further includes citric acid, tannic acid, reserpine, arylpiperazines, alkylaminoquinolones, nocardamines or a pharmaceutically acceptable salt thereof.

[00018] The antibiotic component may further comprise one more betalactamase inhibitor selected from the group comprising sulbactam, tazobactam, clauvulanic acid or a combination thereof.

[00019] In another aspect of the invention a method of of treating one ore more bacterial infection caused by drug-resistant bacterium is provided. The method may include route of administration of selected from the group comprising oral, parenteral (Intravascular IV, Intramuscular IM, Subcutaneous SC), occular, transdermal or combinational administration of an effective amount of a non-antibiotic, non-peptidic entity to a patent under medication of one or more antibiotics in pharmaceutically acceptable dosage form of tablet, capsule, syrup, suspension, solution, powder, granules, gel, lotion, ointment, spray and liposomes or a combination thereof.

[00020] In the method of treatment, the non-antibiotic, non-peptidic entity may be present in a proportion of 0.15 % to about 15% of the antibiotic component.

[00021] The drug-resistant bacterium may be selected from the group comprising ESBL, MRSA, VRSA, VISA, VRE, NDM-1 , MBL producing strain, aminoglycoside resistance strains, efflux over expressed strains, bio-film producing strains, Haemophilus influenzae, Neisseria gonorrhoeaea, N. meningitis, Staph. Epidermidis, pneumococci, Streptococcus mitis, Enterococcus faecalis, Streptococcus pneumoniae, Staphylococcus haemolyticus.

[00022] The bacteria responsible for the infection being treated may at least be Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae, Neisseria gonorrhoeae, Proteus mirabilis, Staphylococcus aureus, A. baumannii, Enterobacter spp., Serratia spp, Serratia marcescens, Streptococcus species, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella ,Moraxella catarrhalis, Streptococcus pyogenes, Streptococcus viridans, Staphylococcus epidermidis, Klebsiella oxytoca, Morganella morganii, Acinetobacter calcoaceticus, Bacteroides fragilis , Peptostreptococcus species, Proteus vulgaris enterococci, Chlamydia trachomatis, Clostridium species, Neisseria meningitidis, Streptococcus agalactiae, Bartonella bacilliformis, Bartonella henselae, Listeria monocytogenes, Tropheryma whipplei, staphylococcus and saprophyticus c.braakii and other species thereof showing resistance to one or more antibiotics. [00023] The bacterial infection being treated may be selected from the group consisting of lower respiratory tract infection, skin and skin structure infection, urinary tract infection, Gonorrhea, Pelvic inflammatory disease, sepsis, soft tissue infections, wound infections,renal infections, nocardial pulmonary infections, mycobacterial lymphadeniti, leprosy, gastrointestinal infections; infections related to abdominal trauma, bloodstream infections, anthrax, plagues; scarlet fever, rheumatic fever, cholera, Haverhill fever, Potomac fever, brucellosis, Carrion's disease, trench fever, bacillary epithelioid angiomatosis, leptospirosis, Lyme disease, rickettsiosis, Q fever, human monocytotropic ehrlichiosis, cat scratch disease, tularemia, pseudo-infections, legionellosis, noscoccomiai infections, erysipeloid, osteomyehtis, prostatitis, peritonitis, encephalitis, cerebrospinal infections, infection of cerebrospinal fluid shunt, meningoencephalitis, joint infections, prosthetic joint infections, septic arthritis, myonecrosis, echyma gangrenosum, cholecystitis, melioidosis, mastoiditis, epididymitis, bursitis.

[00024] The bacterial infection may include infective condition of one or more of disease condition caused by gram negative and gram positive strains and microbes.

[00025] In another aspect of the invention, use of an effective amount of one ore more non-antibiotic, non-peptidic entity for preparation of a medicament for the treatment of infection selected from the group comprising Lower respiratory tract infection, skin and skin structure infection, urinary tract infection, Gonorrhea, Pelvic inflammatory disease, sepsis, soft tissue infections, wound infections,renal infections, nocardial pulmonary infections, mycobacterial lymphadeniti, leprosy, gastrointestinal infections; infections related to abdominal trauma, bloodstream infections, anthrax, plagues; scarlet fever, rheumatic fever, cholera, Haverhill fever, Potomac fever, brucellosis, Carrion's disease, trench fever, bacillary epithelioid angiomatosis, leptospirosis, Lyme disease, rickettsiosis, Q fever, human monocytotropic ehrlichiosis, cat scratch disease, tularemia, pseudo-infections, legionellosis, noscoccomial infections, erysipeloid, osteomyelitis, prostatitis, peritonitis, encephalitis, cerebrospinal infections, infection of cerebrospinal fluid shunt, meningoencephalitis, joint infections, prosthetic joint infections, septic arthritis, myonecrosis, echyma gangrenosum, cholecystitis, melioidosis, mastoiditis, epididymitis, bursitis or a combination thereof is provided.

[00026] During the use, the non-antibiotic, non-peptidic entity may be combined with one or more antibiotic component in a proportion of about 0.15 % to about 1 5% of said antibiotic.

OBJECTS OF THE INVENTION

[00027] In backdrop of the problem looming in th art, a fresh approaches to identify one ore more entity beyond traditional antibiotics or combination of antibiotic are worth investigating that can augment/enhance/potentiate activity of one ore more anti-biotic with regard to antibacterial resistance, antibiotic related side effects and drugs induced toxicities when used together with one or more antibiotic.

[00028] Its is an objective of one of the embodiment of the present invention to disclose one or more natural or synthetic non-antibiotic, non-peptidic compound(s) for prevention of anti microbial resistance of traditional antibiotics and reduction in antibiotic related side effects and drugs induced toxicities.

[00029] Its is an objective of one of the embodiment of the present invention to disclose one or more natural or synthetic non-antibiotic, non-peptidic compound(s) and their synergistic combination with one ore more antibiotic to prevent antimicrobial resistance of traditional antibiotics and reduction in antibiotic related side effects and drugs induced toxicities.

[00030] Another objective of the one of the embodiment of the present invention is to disclose non-antibiotic, non-peptidic compound(s) which when administered with antibiotics reduce multiple drug resistance.

[00031] In another object of the embodiment of the invention, a combination of non- antibiotic, non-peptidic compound(s) or a formulation made thereof enhances or potentates antibacterial activity of any other antibiotic formulation when used together.

[00032] Still another object of the embodiment of the invention is to increase the bioavailabi lity, of the active ingredients of a target antibiotics by co-administering one or more non-antibiotic, non-peptidic compound(s).

[00033] Another object is to disclose compositions of the said formulations.

[00034] Another object of present invention is to enhance the activity and/or spectrum of existing antimicrobial agents .

[00035] Still another object of the invention is to provide enhanced efficacy of an antibiotic formulations and effectiveness even at a low drug concentrations.

[00036] Another object of present invention is to provide safer and better alternative in terms of economic benefits and tolerability for all patients including immuno compromised/old/neonatal patients. [00037] Yet another object of one embodiment of the invention is to provide alternative options which can be formulated with any antibiotic formulations or their solvents and when administered along with antibiotics do not lead to rapid emergence of resistant bacterial strains.

[00038] Yet another object is to disclose one or more non antibiotic compounds alone or in combination reduces toxicity, improves efficacy and reduces treatment time and cost.

BRIEF DESCRIPTION OF DRAWINGS

[00039] The embodiments described herein after would be better understood in terms of their effects, characteristics and mode of operation from the following detailed description with reference to the tables depicting various results wherein::

[00040] Table 1 through Table 3, illustrate in-vitro tests results of antimicrobial activity of individual antibiotic with and without some non-antibiotic, non-peptidic entity(s).

[00041] Table 4 illustrates effect of the non-antibiotic, non-peptidic entity on susceptibility of P. aeruginosa MexAB-OprM-overexpressed and E. coli AcrAB-TolC over expressed strains.

[00042] FIG. 2 (2A-2D) represents effect of EDTA alone and with an antibiotic in eradication of bacterial bio-film development.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[00043] The embodiments herein and the various features and advantageous details thereof are explained more fully with reference to the non-limiting embodiments that are illustrated in the accompanying figures & tables and detailed in the following description. Descriptions of well-known components and processing techniques are omitted so as to not unnecessarily obscure the embodiments herein. The examples used herein are intended merely to facilitate an understanding of ways in which the embodiments herein may be practiced and to further enable those of skill in the art to practice the embodiments herein. Accordingly, the examples should not be construed as limiting the scope of the embodiments herein.

DEFINITION

[00044] The term "non- antibiotic, non-peptidic entity" refers to a compound that has no recognizable antibiotic effect on its own compared to any antimicrobial/antibiotic/ antiviral agent and which is not a polymer of amino acids or fragment of proteins, treated/untreated.

[00045] The term "Effective amount" refers to an amount of an entity sufficient to induce unexpected and/or surprising antimicrobial activity /prevention of antimicrobial resistance/reduction in ADR of a second entity in comparison to the individual activity of either of the entity, renders broader spectrum of activity yet low enough to avoid serious side effects/toxicity.

[00046] The term "microbial resistance/ infection caused by resistant strains" refers to Antimicrobial resistance (AMR) that is resistance of a microorganism to an antimicrobial medicine to which it was previously sensitive. Resistant organisms (they include bacteria, viruses and some parasites), specifically various strains are able to withstand attack by antimicrobial medicines, such as antibiotics, antivirals, and antimalarials, so that standard treatments become ineffective and infections persist and may spread to others. A resistance stain particularly mutates or acquires a resistance gene. Some illustrative class of strains based on their response to corresponding antibiotic includes Antimicrobial resistant Gram-positive bacteria such as Methicillin- resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Enterococcus faecium(VRSA), Vancomycin-resistant Enterococci (VRE), Glycopeptide-intermediate Staphylococcus aureus (GISA), Vancomycin-intermediate Staphylococcus aureus (VISA), New Delhi metallo-beta-lactamase 1(NDM-1) etc. and

Antimicrobial resistant Gram-negative bacteria such as extended spectrum beta lactamase (ESBL) 3rd generation cephalosporin-resistant Escherichia coli , 3rd generation cephalosporin-resistant Klebsiella, Carbapenem-resistant Pseudomonas aeruginosa etc.

[00047] The embodiments of the present invention discloses one or more non-antibiotic, non-peptidic entity(s) of natural or synthetic origin. The non-antibiotic, non-peptidic entity(s) either individually or a combination of one or more of such non-antibiotic, non- peptidic when used together with one or more of conventional antimicrobial agents, provide synergistic activity, combats microbial resistance, enhance the antibacterial effect, reduce toxicity, reduce adverse effects, improve drug safety, reduce treatment time, prevent/ delay resistance development and reduce therapeutic dose of antibiotics.

[00048] The non-antibiotic, non-peptidic entity(s ) is effect at a particular concentration or has a effect at a particular proportion of the antimicrobial agents.

[00049] The non-antibiotic, non-peptidic entity is either present together with the antimicrobial agent or present in a suitable solvent system to₇be used to reconstitute the antimicrobial agent or administered separately along with a specific concentrations according to an embodiment herein.

[00050] The non-antibiotic, non-peptidic entity is/are selected from a group comprising polyamino carboxylic acid, chelating agents, particulate formation inhibitor, stabilizing agent, sodium tetraborate, Sodium salicylate, boric acid, mono phenols, vitamins, minerals, micro nutrients, antioxidants, amino acids, Potassium hydroxide, Phe-Arg-β- naphthylamide, Triclosan, Carotenoids, Phytosterols, Polyphenols including Isoflavones, Flavonones Flavonols, Flavanols, Flavones, Stilbenes, Lignans; Folic acid, Boric acid, Selenium, Lycopene, Lutein, β-Carotene, β-Sitosterol, Genistein, daidzein Naringenin, hesperedin Quercetin, kaemp^'ferol, Catechin, epicatechin, gallocatechin, Luteolin, apigenin ,Resveratrol, Enterodiol, enterolactone, Anthocyanidins, Pelargonidin, malvidin and cyanidin flavonols, flavonones, catechins, lignans, anthroquinone, phytochemical, vitamins, phenolic acids, citric acid, tannic acid, reserpine, arylpiperazines, alkylaminoquinolones, nocardamines or a pharmaceutically acceptable salt thereof according to an embodiment herein.

[00051] The chelating agent is selected from a group comprising ethylene diamine tetraacetic acid (EDTA) and salts thereof, diethylene triamine pentaacetic acid (DTPA), hydroxyethylene diamine triacetic acid (HEDTA), nitrilo triacetic acid (NTA) or a pharmaceutically acceptable salt thereof (normally as a sodium salt) according to an embodiment herein.

[00052] One of the preferred chelating agent is EDTA and salts thereof, preferably edetate disodium as defined in the United States Pharmacopoeia, that can be used with this embodiment.

[00053] In another class of this embodiment without limitation, the non- antibiotic compounds include synthetic/natural amino acids including Larginine, Valine, Leucine, Isoleucine, Alanine,, Glutamine, Lysine, Aspartic acid_* Glutamate, Proline_* Cysteine, Threonine Histidine, Phenylalanine, Tyrosine Tryptophan, Asparagine, Glycine and Serine or a combination thereof.

[00054] In accordance with another embodiments, the non-antibiotic compounds are selected from phytochemicals including natural monophenols such as Apiole, Carnosol, Carvacrol, Dillapiole & Rosemarinol; Flavonols such as Quercetin, Gingerol, aempferol, Myricetin, Rutin & Isorhamnetin; Flavanones such as Hesperidin, Naringenin, Silybin & Eriodictyol; Flavones such as Apigenin, Tangeritin & Luteolin; Flavan-3-ols such as Catechins, (+)-Catechin, (+)-Gallocatechin, (-)-Epicatechin, (-)- Epigallocatechin, (-)-Epigallocatechin gallate (EGCG), (-)-Epicatechin 3-gallate, Theaflavin, Theaflavin-3 -gallate, Theaflavin-3'-gallate, Theaflavin-3,3'-digallate, Thearubigins, Anthocyanins (flavonals) and Anthocyanidins, Pelargonidin , Peonidin, Cyanidin, Delphinidin, Malvidin & Petunidin; Isoflavones (phytoestrogens) such as Daidzein, Genistein, Glycitein, Dihydroflavonols, Chalconoids, Coumestans (phytoestrogens) & Coumestro; Phenolic acids such as Ellagic acid, Gallic acid, Salicylic acid, Tannic acid, Vanillin, Capsaicin, Curcumin, Hydroxycinnamic acids, Caffeic acid, Chlorogenic acid, Cinnamic acid , Ferulic acid and Coumaric acid; Lignans (phytoestrogens) such as Silymarin, Matairesinol, Secoisolariciresinol, Pinoresinol & ariciresinol Terpenes (isoprenoids) and Carotenoids (tetraterpenoids); Carotenes such as β-Carotene, γ-Carotene, δ-Carotene, Lycopene, Neurosporene, Phytofluene & Phytoene; Xanthophylls such as Canthaxanthin, Cryptoxanthin, Zeaxanthin, Astaxanthin, Lutein & Rubixanthin; Triterpenoid such as Oleanolic acid, Ursolic acid, Betulinic acid & Moronic acid; Betalains such as Betacyanins & Betaxanthins) in the form of Indole-3- carbinol, sulforaphane3,3'-Diindolylmethane, Sinigrin, Allicin, Alliin, Allyl isothiocyanate, Piperine & Syn-propanethial-S-oxide.

[00055] In one embodiment, the antimicrobial agent includes one ore more antibiotic(s) component that inhibits or kills growth of bacteria, fungus, protozoa, virus or a combination of them.

[00056] In one embodiment, the antibiotic components are selected from the group comprising beta-lactam antibacterials, which include penicillins, cephalosporins, and carbapenems, aminoglycosides, sulfonamides, quinolones, tetracyclines, macrolides, glycopeptide and oxazolidinones and combinations thereof.

[00057] The antibiotic also include one or more enzyme acting as inhibitor of antibiotic such as betalactamase inhibitor etc. The betalactamase inhibitor is selected from the group comprising sulbactam, tazobactam , clauvulanic acid or a combination thereof.

[00058] The Penicillins is selected from the group comprising penicillin-G ,methicillin, oxacillin,ampicillin, amoxicilin and a combination of either of these with betalactamse inhibitors such as including but not limited to ampicillin+ sulbactam, piperacillin tazobactam, ticarcillin clavulanaic acid, amoxycillin clavulanic acid and the like and pharmaceutically acceptable salts thereof according to an embodiment herein. [00059] In one embodiment, the cephalosporins used include cephalothin, cefazolin, cephapirin, cephradine, cephalexin, cefadroxil, cefaclor, cefamandole, cefonicid, ceforanide, cefuroxime, cefcapene , cefdaloxime , cefditoren , cefetamet , cefixime , cefmenoxime, ceftadizime , cefoperazone , cefotaxime , cefpimizole, cefpodoxime , ceftibuten, ceftriaxone, cefclidine, cefepime, cefluprenam , cefozopran , cefpirome, cefquinome and combination of either of these with other antibitoics or with betalcaatamse inhibitors such as sulbactam, tazobactam or claavulanaic acid or any other class of beta lactamase inhibitor, antibiotic activity inhibitor and pharmaceutically acceptable salts thereof.

[00060] In yet another the embodiment the fluoroquinolone group includes ciprofloxacin, levofloxacin, lomefloxacin ,norfloxacin, parfloxacin, clinafloxacin ,gatifloxacin, ofloxacin and trovafloxacin and pharmaceutically acceptable salts thereof and combination of these with one or more antibiotic drugs .

[00061] In another embodiment, the tetracycline antibiotics include tetracycline doxycycline, minocycline and oxytetracycline, and combination of these with one or more antibiotic drugs or pharmaceutically acceptable salts thereof.

[00062] The Macrolide antibiotics are erythromycin, clarithromycin , azithromycin , dirithromycin, roxithromycin , troleandomycin and pharmaceutically acceptable salts thereof and combination of these with one or more antibiotic drugs according to an embodiment herein.

[00063] In a further embodiment, aminoglycoside groupe includes amikacin, gentamicin, kanamycin, neomycin , streptomycin and tobramycin and pharmaceutically acceptable salts thereof and combination of these with one or more antibiotic drugs.

[00064] It is found and observed that bacteria may be intrinsically resistant to > l class of antimicrobial agents such an antibiotic, or may acquire resistance by de novo mutation or via the acquisition of resistance genes from other organisms. Acquired resistance genes may enable a bacterium to produce enzymes that destroy the antibacterial drug, to express efflux systems that prevent the drug from reaching its intracellular target, to modify the drug's target site, or to produce an alternative metabolic pathway that bypasses the action of the drug. Acquisition of new genetic material by antimicrobial-susceptible bacteria from resistant strains of bacteria may occur through conjugation, transformation, or transduction, with transposons often facilitating the incorporation of the multiple resistance genes into the host's genome or plasmids.

[00065] As represented in Table 1 through Table 4, it has been surprisingly found and observed that different antibiotic such as Amoxicillin (Amoxyclave), cephalosporine (ceftriaxone), carbapenem ( meropenem), amynoglycoside (amikacin), fluoroquinolone ( Ciprofloxacin), macrolide (Azithromycin ) have a significant increase in their efficacy for treatment of infection caused by resistant microorganism when combined with one ore more non-antibiotic, non-peptidic entity compared to their individual potential. The result also shows a significant enhancement in prevention of resistant development. It further suggest that the non-antibiotic, non-peptidic entity do not have any antibacterial activity in their own. However, they potentiate the antibacterial activity of a one ore more accompanying antibiotic, specifically in resistant strain such as P. auriginosa and E.coli.

[00066] FIG. 2A through 2D FIG. 2 (2A-2D) represents SEM image of EDTA alone and with an antibiotic in eradication of bacterial bio-film development which is aone of the mechanism of development of antimicrobial resistance. The SEM images demonstrated that antibiotic ( represented as Ceftriaxone alone or ceftriaxone + sulbactam) with EDTA caused significant disorganization of biofilm and formed a honeycomb like structure of disrupted biofilm and made the cell wall more porous probably by chelating the divalent ions, thus enhancing the entry of drug into the bacterial cells; whereas drugs without EDTA failed to break bacterial biofilm as observed by SEM analysis. FIG. 2A = antibiotic with EDTA ; FIG 2B = antibiotic without EDTA , FIG 2C = antibiotic without EDTA and FIG.2D= antibiotic without EDTA.

[00067] In one embodiment of the invention, the composition is initially in the form of a sterile powder, and wherein the sterile powder is reconstituted prior to administration with a pharmaceutically acceptable solvent.

[00068] It was surprisingly found that the non- antibiotic, non-peptidic entity mentioned above enhances activity of antibiotic compounds against the multi bacterial resistance and help in reducing drug and disease induced toxicities when given in specific weight ratios along with the antibiotic components by one or more of following methods: 1 . Increasing permeability or uptake, synergistic activity; 2. Prevention of efflux; 3. Enzymatic activation; 4. Drug targeting; 5. Preventing loss of enzymes responsible for drug activation; 6.Preventing Transfer of resistant genes by preventing conjugation, transduction etc; 7. Prevention of biofilm and breaking of biofilm; 8. Opening of Porin channels; 9. Reversal of toxin mechanism. Some of this action have been shown in the test results explained herein in-this document.

100069] It is also found and observed that none of the antibiotic components mentioned above have the capacity to deal with all resistance mechanisms and bacteria tend to shift to alternate resistance mechanism in order to survive. Accordingly, it was found by the innovator that the selected non-antibiotic, non-peptidic entity(s) which do not have active recognizable antimicrobial property of their own, utilize one or more mechanisms defined above to overcome bacterial resistance when combined with the antibiotic component, thereby enhancing/ restoring the . activity of antibiotic moiety/compound/combination.

[00070] In a preferred embodiment of the invention, a non-antibiotic, non-peptidic entity to treat infection caused by resistant bacteria and to potentiate activity of one or more antibiotic(s) in one or more microbes/microbiai resistant strain is provided. The entity is selected from the group comprising an effective amount of EDTA disodium, sodium tetraborate, Sodium salicylate, boric acid or a pharmaceutically acceptable salt thereof.

[00071] The non-antibiotic, non-peptidic entity is present in range of about 0. 15 % to about 15% of said antibiotic.

[00072] Each of said non-antibiotic, non-peptidic entity is present in range of about 0.0015 to 1.5% in according to another preferred embodiment herein.

[00073] It is observed and found that the amount of the non-antibiotic, non-peptidic entity do not exceed l Omg/ml or 1 to 1 .5 % of total of a formulation wherein the non- antibiotic, non-peptidic entity is used to augment antibiotic activity of one ore more antibiotic component.

[00074] In yet another embodiment, the microbial resistant strain is selected from the group comprising Extended-spectrum beta-lactamase (ESBL), Methicillin-resistant Staphylococcus aureus (MRSA), Vancomycin-resistant Enterococcus faecium(VRSA), Vancomycin-intermediate Staphylococcus aureus (VISA), Vancomycin-resistant Enterococci (VRE), New Delhi metallo-beta-lactamase l (NDM- l ), metallo-beta- lactamase producing bacteria (MBL) strain, Glycopeptide-intermediate Staphylococcus aureus (GISA), aminoglycoside resistance strains, efflux over expressed strains, bio-film producing strains etc.

[00075] In some embodiment of the invention , the non-antibiotic, non-peptidic entity further comprises citric acid, tannic acid, reserpine, arylpiperazines, alkylaminoquinolones, nocardamines or a pharmaceutically acceptable salt thereof.

[00076] The non-antibiotic, non-peptidic entity is administered either together as a single unit dose with said antibiotic or is administered separately before/after/during administration of said anti-biotic according to an preferred embodiment herein.

[00077] The non-antibiotic, non-peptidic entity is delivered in route selected from the group comprising topical, oral, parenteral and ocular route.

[00078] In a further embodiment of the invention, the non-antibiotic, non-peptidic entity is presented in a form selected from the group comprising nano-particles, dry powder, gel, solution or a pharmaceutically acceptable forms thereof.

[00079] In another preferred aspect of the invention, a synergistic antibiotic composition for treatment of infection caused by resistant bacteria and containment of bacterial resistance is provided. The composition includes a non-antibiotic, non-peptidic entity to potentiate treatment of infection caused by one or more microbial resistant strains and one or more antibiotic component wherein proportion of said non-antibiotic, non- peptidic entity and said antibiotic component is in a range of about 0.15 % to about 15% of said antibiotic. The non-antibiotic, non-peptidic entity is selected from the group comprising an effective amount of EDTA disodium, sodium tetraborate, Sodium salicylate, boric acid or a pharmaceutically acceptable salt thereof.

[00080] The antibiotic component may selected form the group comprising penicillins, cephalosporins, carbapenems, aminoglycosides, sulfonamides, quinolones, tetracyclines, macrolides, glycopeptide , oxazolidinones or combinations thereof.

[00081] The anbitoic component may further include one more betalactamase inhibitor selected from the group comprising sulbactam, tazobactam, clauvulanic acid or a combination thereof.

[00082] The non-antibiotic, non-peptidic entity is present either as a single unit dose with said antibiotic component or is present in a suitable solvent medium according to the above mentioned embodiment.

[00083] The non-antibiotic, non-peptidic entity as mentioned above further includes citric acid, tannic acid, reserpine, arylpiperazines, alkylaminoquinolones, nocardamines or a pharmaceutically acceptable salt thereof.

[00084] The non- antibiotic components may be added to the antibiotic components at the time of administration through a suitable solvent for preparation of reconstitutable solution or may be added with the a antibiotic components at the time of formulation.

[00085] In another preferred aspect of the invention a method of treating bacterial infection caused by a drug-resistant bacterium is provided. The method is selected from the group comprising oral, parenteral (Intravascular IV, Intramuscular IM, Subcutaneous SC), occular, transdermal or combinational administration of the non- antibiotic, non-peptidic entity to a patent under medication of One or more antibiotics in pharmaceutically acceptable dosage form of tablet, capsule, syrup, suspension, solution, powder, granules, gel, lotion, ointment, spray, liposomes and others. The proportion of the non-antibiotic, non-peptidic entity is present in a proportion of 0.15 % to about 15% of the antibiotic component.

[00086] During the treatment, the non-antibiotic, non-peptidic entity is present in a proportion of 0.15 % to about 15% of the antibiotic component.

[00087] The microbes acteria responsible for the infection being treated is at least Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae, Neisseria gonorrhoeae, Proteus mirabilis, Staphylococcus aureus, A. baumannii, Enterobacter spp., Serratia spp, Serratia marcescens, Streptococcus species, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella ,Moraxella catarrhalis, Streptococcus pyogenes, Streptococcus viridans, Staphylococcus epidermidis, Klebsiella oxytoca, Morganella morganii, Acinetobacter calcoaceticus, Bacteroides fragilis , Peptostreptococcus species, Proteus vulgaris enterococci, Chlamydia trachomatis, Clostridium species, Neisseria meningitidis, Streptococcus agalactiae, Bartonella bacilliformis, Bartonella henselae, Listeria monocytogenes, Tropheryma whipplei, staphylococcus and saprophyticus c.braakii.

[00088] The microbes/bacteria infections responsible for bacterial resistance is/are selected from the group consisting of Staphalococcus aureus, Methicillin-resistant Staphylococcus aureus, Multi Drug Resistant Bacteri, Haemophilus influenzae, Neisseria gonorrhoeaea, N. meningitis, Staph. Epidermidis, pneumococci, Streptococcus mitis, Enterococcus faecalis, Streptococcus pneumoniae, Staphylococcus haemolyticus.

[00089] In another embodiment, bacterial infection being treated is selected from the group consisting of Lower respiratory tract infection, skin and skin structure infection, urinary tract infection, Gonorrhea, Pelvic inflammatory disease, sepsis, soft tissue infections, wound infections,renal infections, nocardial pulmonary infections, mycobacterial lymphadeniti, leprosy, gastrointestinal infections; infections related to abdominal trauma, bloodstream infections, anthrax, plagues; scarlet fever, rheumatic fever, cholera, Haverhill fever, Potomac fever, brucellosis, Carrion's disease, trench fever, bacillary epithelioid angiomatosis, leptospirosis, Lyme disease, rickettsiosis, Q fever, human monocytotropic ehrlichiosis, cat scratch disease, tularemia, pseudo- infections, legionellosis, noscoccomial infections, erysipeloid, osteomyelitis, prostatitis, peritonitis, encephalitis, cerebrospinal infections, infection of cerebrospinal fluid shunt, meningoencephalitis, joint infections, prosthetic joint infections, septic arthritis, myonecrosis, echyma gangrenosum, cholecystitis, melioidosis, mastoiditis, epididymitis, bursitis.

[ 00090] In another aspect of the invention, the non-antibiotic, non-peptidic entity alone in combination with one or more antibiotic is used for preparation of a medicament for the treatment of infection selected from the group comprising Lower respiratory tract infection, skin and skin structure infection, urinary tract infection, Gonorrhea, Pelvic inflammatory disease, sepsis, soft tissue infections, wound infections,renal infections, nocardial pulmonary infections, mycobacterial lymphadeniti, leprosy, gastrointestinal infections; infections related to abdominal trauma, bloodstream infections, anthrax, plagues; scarlet fever, rheumatic fever, cholera, Haverhill fever, Potomac fever, brucellosis, Carrion's disease, trench fever, bacillary epithelioid angiomatosis, leptospirosis, Lyme disease, rickettsiosis, Q fever, human monocytotropic ehrlichiosis, cat scratch disease, tularemia, pseudo-infections, legionellosis, noscoccomial infections, erysipeloid, osteomyehtis, prostatitis, peritonitis, encephalitis, cerebrospinal infections, infection of cerebrospinal fluid shunt, meningoencephalitis, joint infections, prosthetic joint infections, septic arthritis, myonecrosis, echyma gangrenosum, cholecystitis, melioidosis, mastoiditis, epididymitis, bursitis. The non-antibiotic, non-peptidic entity is combined with one or more antibiotic component in a proportion of about 0. 1 5 % to about 15% of said antibiotic according to this embodiment herein.

[00091 ] In yet another embodiment of the invention, the composition of non-antibiotic, non-peptidic entity and the antibiotic is administered via/through oral, parenteral (Intravascular IV, Intramuscular IM, Subcutaneous SC), occular, transdermal route or any other suitable route in the pharmaceutically acceptable dosage form of tablet ,capsule, syrup, suspension, solution, powder, granules, gel , lotion , ointment, spray, liposomes, and the like.

[00092] According to another aspect of the present invention, a safe, less toxic alternative in the form of antibiotic component together with non-antibiotic, non- peptidic entity as co-administrable option is provided. This option is having high efficacy against a wide variety of infectious organisms, and to provide compositions that are useful in providing effective treatment against multi drug resistant bacteria.

[00093] According to another aspect of current invention, such combinations have antioxidant activity and help in reducing drug and disease induced toxicities.

[00094] According to yet another aspect addition of non- antibiotic compounds helps in reducing drug resistance and also helps in breaking bacterial resistance.

[00095] Still another aspect is co-administration of non- antibiotic compounds in selective ratios/ ranges, which varies from one antibiotic entity/ molecule to other, improves penetration of antibiotic to the tissues, thereby increasing bio-availability of the drug.

[00096] In another aspect of one of the embodiment of the invention, co-administration of the non-antibiotic, non-peptidic entity in a selected ratios/ ranges, helps in targeting one or more antibiotic components or combination thereof, thereby reducing dose of antibiotics.

[00097] In yet another aspect of the embodiment of the invention, the above mentioned composition is presented in a pharmaceutically administrable form or are present as a kit along with the antibiotics.

[00098] In yet another embodiment of the invention, the non-antibiotic, non-peptidic components, in defined concentrations, is mixed in API during manufacturing.

[00099] In another aspect of the invention, different formulation were prepared making specific selection from the different elements of the antibiotic components and non- antibiotic, non-peptidic elements in specific concentrations.

[000100] The non-antibiotic, non-peptidic entity alone or a combination of selected entity can be formulated as a single unit, packed in a sealed air tight pharmaceutically ^•acceptable container. The container is selected from the group consisting of vial, an ampoule, a syringe, a packet, a pouch, an autoinjecter, as a solvent system, in tube, patch and the like, present as liquid, gel, emulsions, colloidals, dry powder for reconstitution, lyophilized powder, as a dose concentrate in the form of drug particles, powders, granules, nano-particles, microspheres and any other pharmaceutically administrable forms and the like. The final composition/formulation is suitably protected from moisture, light and degradation with the help of vacuum or inert gas micro atmosphere or in pressurized carbon dioxide according to an embodiment herein.

[000101] Above disclosure describe a manner and method of making using the invention and sets forth the best mode contemplated by the inventor for carrying out his invention but is not to be construed as limiting. Various modifications and variations of the described method and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention should not be unduly limited to such specific embodiments. Indeed, various modifications and equivalents of the described modes for carrying out the invention that are obvious to those skilled in formulation development or related fields are intended to be within the scope of the invention. The foregoing description of the specific embodiments will so fully reveal the general nature of the embodiments herein that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific embodiments without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments herein have been described in terms of preferred embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the spirit and scope therein.

Claims

We Claim:

1 . A non-antibiotic, non-peptidic entity to potentiate activity of one or more antibiotic(s) in

treatment of infection(s) caused by one or more microbial resistant strain(s),

wherein said entity is selected from the group comprising an effective amount of EDTA disodium, sodium tetraborate, Sodium salicylate, boric acid or a pharmaceutically acceptable salt thereof.

2. The non-antibiotic, non-peptidic entity of claim 1 , wherein said entity is present in range of about 0.15 % to about 15% of said antibiotic.

3. The non-antibiotic, non-peptidic entity of claim 1 , wherein each entity is present in range of about 0.0015 to 1.5%.

4. The non-antibiotic, non-peptidic entity of claim 1 , wherein said microbial resistant strain is selected from the group comprising ESBL, MRSA, VRSA, VISA, VRJE, NDM- 1 , MBL producing strain, aminoglycoside resistance strains, efflux over expressed strains, bio-film producing strains or a combination thereof,.

5. The non-antibiotic, non-peptidic entity of claim 1 , wherein said non-antibiotic, non-peptidic entity further comprises citric acid, tannic acid, reserpine, arylpiperazines,

alkylaminoquinolones, nocardamines or a pharmaceutically acceptable salt thereof.

6. The entity of claim 1 , wherein said entity is administered either together as a single unit dose with said antibiotic or is administered separately before/after/during administration of said antibiotic.

7. The entity of claim 1 , wherein said entity is delivered in route selected from the group

comprising topical, oral, parenteral and ocular route.

8. The entity of claim 1 , wherein said entity is presented in a form selected from the group

comprising nano-particles, dry powder, gel, solution or a pharmaceutically acceptable forms thereof.

9. A synergistic antibiotic composition for treatment and containment of bacterial resistance, said composition comprising :

a non-antibiotic, non-peptidic entity to potentiate activity of one or more antibiotic(s) component in treatment of infection(s) caused by one or more microbial resistant strain(s), wherein said entity is selected from the group comprising an effective amount of EDTA disodium, sodium tetraborate, Sodium salicylate, boric acid or a pharmaceutically acceptable salt thereof;

wherein said antibiotic component is selected form the group comprising penicillins, cephalosporins, carbapenems, aminoglycosides, sulfonamides, quinolones, tetracyclines, macrolides, glycopeptide , oxazolidinones or combinations thereof; and

wherein proportion of said non-antibiotic, non-peptidic entity and said antibiotic component is in a range of about 0.15 % to about 15% of said antibiotic.

10. The synergistic composition of claim 4, wherein said non-antibiotic, non-peptidic entity is

present either as a single unit dose with said antibiotic component or is present in a suitable solvent medium.

11. The synergistic composition of claim 4, wherein said non-antibiotic, non-peptidic entity further comprising citric acid, tannic acid, reserpine, arylpiperazines, alkylaminoquinolones, nocardamines or a pharmaceutically acceptable salt thereof.

12. The synergistic composition of claim 4, wherein said antibiotic component further comprises one more betalactamase inhibitor selected from the group comprising sulbactam, tazobactarn, Λ

clauvulanic acid or a combination thereof.

13. A method of of treating one ore more bacterial infection caused by drug-resistant bacterium, comprising route of administration of selected from the group comprising oral, parenteral (Intravascular IV, Intramuscular IM, Subcutaneous SC), occular, transdermal or combinational administration of an effective amount of a non-antibiotic, non-peptidic entity to a patent under medication of one or more antibiotics in pharmaceutically acceptable dosage form of tablet, capsule, syrup, suspension, solution, powder, granules, gel, lotion, ointment, spray and liposomes or a combination thereof.

14. The method of treatment of claim 13, wherein said non-antibiotici non-peptidic entity is present in a proportion of 0.15 % to about 15% of the antibiotic component.

15. The method of treatment of claim 13, drug-resistant bacterium is selected from the group comprising ESBL, MRSA, VRSA, VISA, VRE, NDM- 1 , MBL producing strain, aminoglycoside resistance strains, efflux over expressed strains, bio-film producing strains, Haemophilus influenzae, Neisseria gonorrhoeaea, N. meningitis, Staph. Epidermidis, pneumococci, Streptococcus mitis, Enterococcus faecalis, Streptococcus pneumoni; Staphylococcus haemolyticus.

16. The method of treatment of claim 13, wherein said bacteria responsible for the infection bei treated is at least Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumonii Streptococcus pneumoniae, Haemophilus influenzae, Neisseria gonorrhoeae, Proteus mirabil Staphylococcus aureus, A. baumannii, Enterobacter spp., Serratia spp, Serratia marcescei Streptococcus species, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legione ,Moraxella catarrhalis, Streptococcus pyogenes, Streptococcus viridans, Staphylococc epidermidis, Klebsiella oxytoca, Morganella morganii, Acinetobacter calcoaceticus, Bacteroic fragilis , Peptostreptococcus species, Proteus vulgaris enterococci, Chlamydia trachomat Clostridium species, Neisseria meningitidis, Streptococcus agalactiae, Bartonella bacilliform Bartonella henselae, Listeria monocytogenes, Tropheryma whipplei, staphylococcus a saprophyticus c.braakii and other species thereof showing resistance to one or more antibiotic:

17. The method of treatment of claim 13, wherein said bacterial infection being treated is select from the group consisting of lower respiratory tract infection, skin and skin structure infectk urinary tract infection, Gonorrhea, Pelvic inflammatory disease, sepsis, soft tissue infectioi wound infections,renal infections, nocardial pulmonary infections, mycobacterial lymphaden leprosy, gastrointestinal infections; infections related to abdominal trauma, bloodstrei infections, anthrax, plagues; scarlet fever, rheumatic fever, cholera, Haverhill fever, Potom fever, brucellosis, Carrion's disease, trench fever, bacillary epithelioid angiomatos leptospirosis, Lyme disease, rickettsiosis, Q fever, human monocytotropic ehrlichiosis, < scratch disease, tularemia, pseudo-infections, legionellosis, noscoccomial infectioi erysipeloid, osteomyehtis, prostatitis, peritonitis, encephalitis, cerebrospinal infectioi infection of cerebrospinal fluid shunt, meningoencephalitis, joint infections, prosthetic jo infections, septic arthritis, myonecrosis, echyma gangrenosum, cholecystitis, 'melioidos mastoiditis, epididymitis, bursitis.

18. The method of treatment of claim 13, wherein said bacterial infection comprises infecti condition of one or more of disease condition caused by gram negative and gram positi strains and microbes.

19. Use of an effective amount of one ore more non-antibiotic, non-peptidic entity for preparati of a medicament for the treatment of infection selected from the group comprising Low respiratory tract infection, skin and skin structure infection, urinary tract infection, Gonorrhi Pelvic inflammatory disease, sepsis, soft tissue infections, wound infections,renal infectioi nocardial pulmonary infections, mycobacterial lymphadeniti, leprosy, gastrointestir infections; infections related to abdominal trauma, bloodstream infections, anthrax, plagu< scarlet fever, rheumatic fever, cholera, Haverhill fever, Potomac fever, brucellosis, Carrioi disease, trench fever, bacillary epithelioid angiomatosis, leptospirosis, Lyme disea: rickettsiosis, Q fever, human monocytotropic ehrlichiosis, cat scratch disease, tularem pseudo-infections, legionellosis, noscoccomial infections, erysipeloid, osteomyehtis, prostatit peritonitis, encephalitis, cerebrospinal infections, infection of cerebrospinal fluid shu meningoencephalitis, joint infections, prosthetic joint infections, septic arthritis, myonecros echyma gangrenosum, cholecystitis, melioidosis, mastoiditis, epididymitis, bursitis or combination thereof.

20. The use of claim 19, wherein said non-antibiotic, non-peptidic entity is combined with one more antibiotic component in a proportion of about 0.15 % to about 15% of said antibiotic.