Method of treating bacterial infections

Abstract

The present invention relates to a composition comprising a first compound which is a nucleoside analogue capable of inhibiting a bacterial colonisation or infection of a subject; a second compound which is capable of decreasing mitochondrial toxicity of said nucleoside analogue and surprisingly enhance the antibacterial effect of the combination; and a third compound capable of decreasing the concentration in bacteria of nucleosides and/or nucleotides known to compete with nucleoside analogues. The first compound may be AZT, FdUrd, 5-fluorouracil, BrdUrd, IdUrd, didanosine or gemcitabine; the second compound may be uridine or a uridine-comprising compound; and the third compound may be iclaprim; trimethoprim; or a compound comprising trimethoprim, such as trimethoprim-sulfa.

Claims

1. A method of enhancing the antibacterial effect in a subject of at least one nucleoside analogue, which method comprises administering an effective amount of said at least one nucleoside analogue to a subject in need thereof in combination with a) iclaprim and b) at least one second compound selected from the group consisting of uridine, cytidine and prodrugs thereof; wherein said nucleoside analogue is selected from the group consisting of AZT, FdUrd, gemcitabine, didanosine, 5-fluorouracil, 5-bromouracil, 5-iodouracil (IdUrd), FLT, flucytosine, 5-bromouridine (BrUrd), stavudine, telbivudine, entecavir, emtricitabine, adefovir, lamivudine, tenofovir, vidarabine, abacavir, cytarabine, aracytidine, cidofovir, zalcitabine, ribavirin, idoxuridine, trifluoruridine, mercaptopurine, 5-ethyldeoxyuridine, thioguanine, 5-chlorodeoxyuridine, pentostatin, cladribine, clofarabine, fludarabine, nalarabine, allopurinol, FMdc, sinefungin, troxacitabine, vidaza, nelarabine, decitabine, CNDAC, ECyd, sofosbuvir, sapacitabine, mericitabine, amdoxovir, apricitabine, telbivudine, clevudine, bestifovir, brincidofovir, nicavir, and prodrugs thereof.

2. The method according to claim 1, wherein the toxicity of said at least one nucleoside analogue is reduced by the at least one second compound which also enhances the antibacterial effect.

3. The method according to claim 1, wherein the combined antibacterial effect of the nucleoside analogue, iclaprim and the second compound is greater than the sum of the effect provided if administered separately and the toxicity is reduced.

4. The method according to claim 1, wherein the nucleoside analogue is selected from the group consisting of AZT; FdUrd; 5-fluorouracil; BrdUrd; IdUrd; didanosine and gemcitabine.

5. The method according to claim 4, wherein the nucleoside analogue is AZT and the second compound is uridine.

6. A method according to claim 4, wherein the nucleoside analogue is FdUrd and the second compound is uridine.

7. A method according to claim 4, wherein the nucleoside analogue is didanosine and the second compound is uridine.

8. A method according to claim 4, wherein the nucleoside analogue is BrdUrd and the second compound is uridine.

9. The method according to claim 1, wherein the subject is a mammal.

10. A composition comprising as a first compound at least one nucleoside analogue which is capable of decreasing a bacterial colonisation or infection of a subject; a second compound which is capable of decreasing mitochondrial toxicity of said at least one nucleoside analogue while at the same time increases its antibacterial effect; and a third compound capable of decreasing the concentration in bacteria of nucleosides and/or nucleotides known to compete with nucleoside analogues, wherein said third compound is iclaprim; wherein said nucleoside analogue is selected from the group consisting of AZT, FdUrd, gemcitabine, didanosine, 5-fluorouracil, 5-bromouracil, 5-iodouracil (IdUrd), FLT, flucytosine, 5-bromouridine (BrUrd), stavudine, telbivudine, entecavir, emtricitabine, adefovir, lamivudine, tenofovir, vidarabine, abacavir, cytarabine, aracytidine, cidofovir, zalcitabine, ribavirin, idoxuridine, trifluoruridine, mercaptopurine, 5-ethyldeoxyuridine, thioguanine, 5-chlorodeoxyuridine, pentostatin, cladribine, clofarabine, fludarabine, nalarabine, allopurinol, FMdc, sinefungin, troxacitabine, vidaza, nelarabine, decitabine, CNDAC, ECyd, sofosbuvir, sapacitabine, mericitabine, amdoxovir, apricitabine, telbivudine, clevudine, bestifovir, brincidofovir, nicavir, and prodrugs thereof; and wherein said second compound is selected from the group consisting of uridine, cytidine and prodrugs thereof.

11. A composition according to claim 10, wherein the first compound is selected from the group consisting of AZT, FdUrd, 5-fluorouracil, BrdUrd, IdUrd, didanosine, gemcitabine and prodrugs giving rise to these compounds.

12. A composition according to claim 10, wherein the second compound is a compound selected from the group consisting of uridine, cytidine and prodrugs giving rise to these compounds.

13. A composition according to claim 10, in combination with a carrier suitable for use in a medicament.

14. A pharmaceutical preparation comprising at least one first compound selected from the group consisting of AZT, FdUrd, 5-fluorouracil, BrdUrd, IdUrd, didanosine and gemcitabine; at least one second compound selected from the group consisting of uridine, cytidine, and prodrugs giving rise to these compounds; and at least one third compound which is iclaprim; or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier.

15. The pharmaceutical preparation according to claim 14, wherein the first compound is AZT and the second compound is uridine.

16. The pharmaceutical preparation according to claim 14, which is in liquid form.

17. The pharmaceutical preparation according to claim 14, which is in a solid form suitable for oral administration.

18. A method of treating or preventing a bacterial infection or colonisation in a subject, comprising administering a therapeutically efficient amount of at least one first compound selected from the group consisting of AZT, FdUrd, 5-fluourouracil, BrdUrd, IdUrd, didanosine and gemcitabine; at least one second compound selected from the group consisting of uridine, cytidine and prodrugs thereof; and at least one third compound which is iclaprim, to said subject.

19. A method according to claim 18, wherein said at least one second compound selected from the group consisting of uridine, cytidine and prodrugs thereof is co-administered with the at least one first compound and iclaprim.

20. A method according to claim 18, wherein the antibacterial nucleoside analogue is selected from the group consisting of AZT; FdUrd; 5-fluorouracil; BrdUrd; IdUrd; didanosine and gemcitabine.

21. The method according to claim 18, wherein the bacteria is Gram-negative and Gram-positive bacteria selected from the group consisting of Acinetobacter; Bacillus; Bordetella; Campylobacter; Chlamidia, Citrobacter; Clostridium; Corynebacteria; Enterobacter; Escherichia; Haemophilus; Heliobacter; Klebsiella; Legionella; Listeria; Micrococcus; Moraxella; Mycobacteria; Mycoplasma; Neisseria; Proteus; Pseudomonas; Salmonella; Serratia, Shigella; Staphylococcus; Streptococcus; Vibrio and Yersinia.

22. A method according to claim 18, wherein the subject is a mammal, and the method provides an enhanced potency of the nucleoside as well as a decreased toxicity to the subject as compared to the result of separate administration of said compounds.

23. The method according to claim 4, wherein the nucleoside analogue is gemcitabine and the second compound is uridine.

24. The method according to claim 1, wherein the nucleoside analogue is selected from the group consisting of AZT; FdUrd; didanosine and gemcitabine.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) As appears from the above, a combination of AZT and uridine has been found not only to retain the broad antibacterial effect of AZT against multidrug resistant bacteria, but uridine also enhances the antibacterial effect of AZT, while at the same time uridine reduces toxicity to animal cells. AZT+uridine are combined with trimethoprim, iclaprim or compounds with similar effects, as shown in the experimental part below, to achieve a synergistic antibacterial effect, while retaining a decreased toxicity.

(2) In a first aspect, the invention relates to a method of enhancing an antibacterial effect in a subject of a nucleoside analogue commonly used against viral infections and/or in cancer treatment, as discussed above, while at the same time reducing toxicity.

(3) More specifically, one embodiment of the invention is a method of enhancing the antibacterial effect in a subject of a nucleoside analogue, which method comprises administering an effective amount of at least one nucleoside analogue to a subject in need thereof in combination with at least one second compound selected from the group consisting of uridine and uridine-comprising compounds; and at least one third compound selected from the group consisting of trimethoprim and/or other trimethoprim compounds. In this context, it is to be understood that the term trimethoprim compounds includes trimethoprim-comprising compounds as well as prodrugs capable of giving rise to trimethoprim or such compounds. The term trimethoprim-comprising compounds is understood herein to mean any compound which comprises a sufficient amount of trimethoprim to provide an equivalent or close to equivalent effect to trimethoprim in the herein described method and use.

(4) The second compound may be a compound known to reduce the toxicity of a nucleoside analogue, such as uridine and/or uridine-comprising compounds. In this context, the term uridine-comprising compounds is understood to mean any compound including cytidine and cytidine prodrugs which comprises a sufficient amount of uridine to provide an equivalent or close to equivalent effect to uridine in the herein described method and use. As will appear from the Experimental part below, such as in Table 9A, according to the present invention, the second compound will also enhance the antibacterial effect of a nucleoside analogue, which has presently been used to treat viral infections or cancer.

(5) Nucleoside analogues commonly used as drugs and used according to the invention may be one or more compound selected from the group consisting of AZT, FdUrd, gemcitabine, didanosine, 5-fluorouracil, 5-bromouracil, 5-iodouracil, FLT, flucytosine, 5-bromouridine, stavudine, telbivudine, entecavir, emtricitabine, adefovir, lamivudine, tenofovir, vidarabine, abacavir, cytarabine, aracytidine, cidofovir, zalcitabine, ribavirin, idoxuridine, trifluoruridine, mercaptopurine, 5-ethyldeoxyuridine, thioguanine, 5-chlorodeoxyuridine, pentostatin, cladribine, clofarabine, fludarabine, nalarabine, allopurinol, FMdc, sinefungin, troxacitabine, vidaza, nelarabine, decitabine, CNDAC, ECyd, sofosbuvir, sapacitabine, mericitabine, amdoxovir, apricitabine, telbivudine, clevudine, bestifovir, brincidofovir and nicavir. As the skilled person will appreciate, prodrugs of such compounds may also be useful according to the invention to improve oral bioavailability and/or penetration and/or uptake into bacteria.

(6) In one embodiment, the nucleoside analogue is selected from the group consisting of AZT, FdUrd, 5-fluorouracil, BrdUrd, IdUrd, didanosine and gemcitabine.

(7) The compound inhibiting tetrahydrofolate synthesis may be iclaprim or trimethoprim e.g. trimethoprim-sulfa. As the skilled person will appreciate, other trimethoprim-containing compounds may be useful provided the advantageous effect of enhancing the antibacterial effect is obtained.

(8) In an advantageous embodiment of the method according to the invention, the combined antibacterial effect of the nucleoside analogue and the second and third compound is greater than the sum of the effect provided if administered separately. In other words, a synergistic effect has been observed, as will be illustrated by the Experimental part below.

(9) In a specific embodiment of the first aspect, the nucleoside analogue is AZT or FdUrd, the second compound is uridine and the third compound is trimethoprim or iclaprim.

(10) The subject may be a human or an animal, as will be discussed in more detail below, and the antibacterial effect may be directed towards a bacterial infection caused by Gram-negative and/or Gram-positive bacteria, especially by bacteria which have acquired resistance to antibiotics in present use.

(11) In a second aspect, the invention relates to a composition comprising as a first compound at least one nucleoside analogue capable of inhibiting a bacterial colonisation or infection of a subject; at least one second compound capable of decreasing mitochondrial toxicity of said nucleoside analogue and increasing the antibacterial effect; and at least one third compound capable of decreasing the concentration in bacteria of nucleosides and/or nucleotides known to compete with nucleoside analogues.

(12) The first compound may be one or more of the nucleoside analogues commonly used as drugs, as discussed above in relation to the first aspect of the invention. In one embodiment, the first compound is selected from the group consisting of AZT, FdUrd, BrdUrd, IdUrd, didanosine and gemcitabine.

(13) The second compound may be a compound as discussed above in relation to the first aspect of the invention, such as a compound commonly known to reduce the toxicity and surprisingly increase the antibacterial effect of the first compound, and hence allow larger doses, of nucleoside analogue-based drugs. In one embodiment of the composition, the second compound is uridine.

(14) The third compound may be a compound as discussed above in relation to the first aspect of the invention, such as a folate analogue, such as trimethoprim, or a compound comprising trimethoprim such as trimethoprim-sulfa, any other trimethoprim compound, or a compound inhibiting bacterial synthesis of tetrahydrofolate. In one embodiment, the third compound is iclaprim or trimethoprim and/or other trimethoprim compound(s).

(15) The compound according to the invention is the first medical use of a composition comprising at least the herein defined three components. Thus, the present invention relates to a composition according to the invention for use as a medicament.

(16) Further, as appears from the present application as a whole, the present composition comprising at least the herein defined three components is suggested for the first time as an antibacterial composition. Thus, the present invention also relates to a composition according to the invention for use in the treatment or prevention of bacterial infections. It will appear from the Experimental part below which bacteria the present composition is advantageously used against, and the skilled person in this area is well aware of which diseases and/or conditions they may cause.

(17) A specific aspect of the invention is a composition according to the invention which is a pharmaceutical preparation. Such a pharmaceutical preparation comprises at least one first compound selected from the group consisting of AZT, FdUrd, 5-fluorouracil, BrdUrd, IdUrd, didanosine and gemcitabine; at least one second compound selected from the group consisting of uridine and uridine-comprising compounds; and at least one third compound selected from the group consisting of trimethoprim; trimethoprim compounds, such as trimethoprim-sulfa; and iclaprim together with a pharmaceutically acceptable carrier.

(18) The present invention also encompasses any pharmaceutically acceptable salt of the above, wherein the biological effectiveness and properties of the compounds of the present disclosure are retained.

(19) Further, the pharmaceutical composition of the invention may comprise an excipient selected from the group comprising, but not limited to, binders and compression aids, coatings and films, colouring agents diluents and vehicles disintegrants, emulsifying and solubilising agents, flavours and sweeteners, repellents, glidants and lubricants, plasticisers, preservatives, propellants, solvents, stabilisers, suspending agents and viscosity enhancers. The pharmaceutical composition of the invention may be administered in a variety of unit dosages depending on the method of administration, target site, physiological state of the subject, and other medicaments administered.

(20) As the skilled person will appreciate, in severely sick patients, intravenous dosing with fixed combinations is preferred, and the compositions of the invention may alternatively be formulated for delivery by injection. As an example, the compound is delivered by injection by any one of the following routes: intravenous, intramuscular, intradermal, intraperitoneal or subcutaneous route.

(21) Thus, in one embodiment, the compositions described are liquid formulations, such as liquids suitable for intravenous administration. The skilled person will be capable of preparing such formulations based on the common general knowledge of the field and assisted by common textbooks and publications in the area.

(22) Examples of liquid compositions include solutions, emulsions, injection solutions, and solutions contained in capsules. Generally, any solvent that has the desired effect and can be administered to a subject may be used. The solvent may be a pure solvent or may be a mixture of liquid solvent components. In some variations the solution formed is an in situ gelling formulation. Solvents and types of solutions that may be used are well known to those versed in drug delivery technologies.

(23) The composition described herein may be in the form of a liquid suspension. The liquid suspensions may be prepared according to standard procedures known in the art. Examples of liquid suspensions include micro-emulsions, the formation of complexing compounds, and stabilising suspensions. The liquid suspension may be in diluted or concentrated form. Liquid suspensions for oral use may contain suitable preservatives, antioxidants, and other excipients known in the art functioning as one or more of dispersion agents, suspending agents, thickening agents, emulsifying agents, wetting agents, solubilising agents, stabilising agents, flavouring and sweetening agents, colouring agents, and the like. The liquid suspension may contain glycerol and water.

(24) For less severely sick patients, or for preventive purposes, the composition according to the invention may advantageously be administered orally, such as in solid dosage forms such as powder, tablets, pills, and capsules, or in liquid dosage forms, such as oral pastes, elixirs, syrups, solutions and suspensions.

(25) Thus, in one embodiment, the pharmaceutical composition of the invention may be formulated for oral administration. Traditional inactive ingredients may be added to provide desirable colour, taste, stability, buffering capacity, dispersion, or other known desirable features. Conventional diluents may be used to make compressed tablets. Both tablets and capsules may be manufactured as sustained-release compositions for the continual release of medication over a period of time. Compressed tablets may be in the form of sugar coated or film coated tablets, or enteric-coated tablets for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration may contain colouring and/or flavouring to increase patient compliance.

(26) Aerosol formulations can be used for patients with lung infections, such as cystic fibrosis and COPD, where uridine or an uridine-comprising component may be beneficial also to treat the underlying chronic disease.

(27) Dosing can be done by giving compounds such as AZT, FdUrd, 5-flourouracil, BrdUrd, IdUrd, didanosine or gemcitabine combined in one separate formulation with compounds such as iclaprim, trimethoprim, trimethoprim-sulfa and/or other trimethoprim compounds and uridine and/or uridine-comprising compounds in one or more separate formulation(s).

(28) As appears from the above, the invention also encompasses the use of a compound according to the invention, or therapeutically acceptable salts thereof, in the manufacture of medicaments for the treatment of a bacterial colonisation or infection in a subject. With regard to general doses and forms of administration the compounds used in the present invention, reference is made e.g. to Gahart's 2016, 32.sup.nd Edition: Intravenous medications, A Handbook for Nurses and Health Professionals (Elsevier.com). The present invention also includes a method of inhibiting a bacterial colonisation or infection of a host, which method includes contacting the bacteria with an effective amount of an antibacterial nucleoside analogue in combination with uridine or a uridine-comprising compound and an effective amount of at least one iclaprim or trimethoprim compound, wherein the concentration of nucleosides and/or nucleotides competing with the nucleoside analogue in bacteria or parasites is reduced while mitochondria in the colonised or infected area of said host remains substantially uneffected.

(29) Examples of compounds decreasing nucleoside/nucleotide levels in bacteria but not in mitochondria are didox, trimidox, metronidazole, rifampicin, brodimoprim, aditoprim trimethoprim, iclaprim, sulfa, sulfonamides, gemcitabine, FMdC, clofarabine, methotrexate, mercaptopurine, para-amino-salicylic acid, inhibitors of bacterial ribonucleotide reductase and dihydrofolate reductase, and prodrugs of such compounds to improve oral bioavailability and/or penetration/uptake into bacteria.

(30) Further examples of third compounds that may be combined with a first and a second compound according to the invention are e.g. pyrimethamine, dapsonpyrimethamine, pyrimethamine-sulfadoxine (Fansidar), diaminodiphenyl sulfone (Dapsone), trimetrexate, proguanil and pemetrexel. In one embodiment, uridine is co-administered with the nucleoside analogue and trimethoprim or a compound comprising iclaprim or trimethoprim, such as trimethoprim-sulfa.

(31) In one embodiment, the antibacterial nucleoside analogue is selected from the group consisting of AZT; FdUrd; 5-flurouracil, BrdUrd, IdUrd, didanosine and gemcitabine.

(32) In a third aspect, the invention relates to a method of treating or preventing a bacterial infection or colonisation in a subject, which method comprises administering to said subject an effective dose of at least one first compound selected from the group consisting of AZT, FdUrd, 5-fluourouracil, BrdUrd, IdUrd, didanosine and gemcitabine; at least one second compound selected from the group consisting of uridine and uridine-comprising compounds; and a third compound selected from the group consisting of trimethoprim ;trimethoprim compounds; and iclaprim.

(33) All details, discussions and examples of the first, second and third compounds provided above in the context of the first and the second aspect of the invention will also apply to the third aspect.

(34) The skilled person in this field will be aware of commonly used doses of the compounds used in combination according to the invention, and will therefore be capable of prescribing the appropriate doses and/or amounts used for a given context.

(35) Thus, the method according to the invention may include daily infusion of AZT as the first compound at a dose of 1.5-15 mg/kg; daily infusion of uridine as the second compound at a dose of 30-300 mg/kg; and as a third compound trimethoprim at a dose of 1.5-15 mg /kg, or iclaprim at a dose of 0.5-5 mg/kg. Such treatment may include 4-6 doses daily of each compound.

(36) Further, the method according to the invention may include daily oral administration of AZT as the first compound at a dose of 2-20 mg/kg and daily oral administration of uridine or uridine prodrug as the second compound at a dose of 60-600 mg/kg and as the third compound trimethoprim at a dose of 2-20 mg/kg or iclaprim at a dose of 1-10 mg/kg. Such treatment may include 1-4 daily doses.

(37) If AZT or other nucleosides are given as prodrugs, such as 5-O-aminoacid derivatives, an enhanced antibacterial effect can be achieved or a reduced dose given.

(38) If AZT or other nucleosides are given as phosphonate prodrugs, such as nicavir and M2 (compound 2 below), an enhanced antibacterial effect, a reduced dose given or a different resistance pattern may be achieved.

(39) ##STR00001##

(40) Further illustrative doses will be presented in the Experimental part below, and the skilled person will be able to design suitable dose schemes and regimes based on this specification as a whole together with common general knowledge.

(41) The bacteria may be Gram-negative and/or Gram-positive bacteria, which bacteria may or may not be anaerobic, such as Acinetobacter; Bacillus; Bordetella; Campylobacter; Chlamydia; Citrobacter; Clostridium; Corynebacteria; Enterobacter; Enterococcus; Escherichia; Haemophilus; Helicobacter; Klebsiella; Legionella; Listeria; Micrococcus; Moraxella; Mycobacteria; Mycoplasma; Neisseria; Proteus; Pseudomonas; Salmonella; Serratia; Shigella; Staphylococcus; Streptococcus; Vibrio and Yersinia. Some relevant bacteria will be further illustrated in the Experimental part below, where publically available strains have been tested, as marked or noted e.g. by reference to the relevant depository such as ATCC.

(42) The subject receiving treatment in accordance with the present invention may be any subject capable of colonisation and infection by bacteria, as appears from below.

EXPERIMENTAL PART

(43) The examples below are provided for illustrative purposes only, and should not be construed as limiting the invention as defined by the appended claims. All references provided below or elsewhere in the present application are hereby included herein via reference. The present invention makes use of, unless otherwise indicated, conventional microbiological techniques within the skill of the art. Such conventional techniques are known to the skilled worker.

Example 1

Bacterial Strains Tested

(44) Table 1 below shows some of the bacterial strains used in studies to test the antibacterial effect.

(45) TABLE-US-00001 TABLE 1 Working Pathogen Denotation code no Properties Escherichia coli (S) CCUG 24T 1 Type strain Escherichia coli (R) LMG 15862 2 Penicillin resistant strain Acinetobacter baumannii (S) LMG 1041T 3 Type strain Escherichia coli (S) LMG 8223 5 Enterobacter cloacae (S) LMG 2783T 6 Type strain Klebsiella pneumoniae (S) CCUG 225T 7 Type strain Klebsiella pneumoniae (R) LMG20218 8 ESBL Pseudomonas aeruginosa (S) LMG 6395 10 Staphylococcus aureus (S/R) LMG 10147 12 MSSA/weak penicillinase producer Staphylococcus aureus (R) LMG 15975 13 MRSA (S)Strains sensitive to antibiotics; (R)Strains resistant to antibiotics BCCM/LMGstrains obtained from Belgian Coordinated Collection of Microorganisms CCUGstrains obtained from Culture Collection of University of Goteborg

Example 2

Antibacterial Effect of AZT Combined with Uridine

(46) Examples of the antibacterial effect of the combination AZT and uridine against bacteria are given in Table 2 below. More specifically, the Minimal Inhibitory Concentration (MIC, g/ml) of AZT against a range of human pathogens in the presence of varying concentrations of uridine is provided.

(47) TABLE-US-00002 TABLE 2 Uridine concentration (g/ml) Bacterial pathogen 0 1 10 50 100 200 E. coli strain no 2 >20 25 >20 25 >20 25 >15 20 >5 15 >5 15 E. coli strain no 1 >0.5 1 nt >0.5 1 >0.5 1 >0.5 1 >0.5 1 E. coli strain no 5 >4 8 nt nt nt nt nt K. pneumoniae strain no 8 >15 25 >10 20 >10 20 >10 20 >10 20 >10 20 K. pneumoniae strain no 7 >1 2 nt nt nt nt nt E. cloacae strain no 6 >10 20 nt >5 10 >5 10 >2 5 >2 5 A. baumannii strain no 3 >150 nt >150 >150 >150 >150 P. aeruginosa strain no 10 >150 nt >150 >150 >150 >150 S. aureus strain no 13 >150 nt >150 >150 >150 >150

(48) AZT was active against E. coli, K. pneumonia and E. cloacae. Addition of up to 200 g/ml of uridine surprisingly resulted in an increase in antibacterial effect, in contrast to the previously reported decrease in antibacterial effect by addition of thymidine.

Example 3

Synergistic Effect of AZT Combined with Uridine and Trimethoprim

(49) In order to further increase the antibacterial effect of AZT, or other nucleoside analogues including prodrugs, trimethoprim, trimethoprim+sulfa, or other compounds decreasing the concentration of nucleosides/nucleotides in bacteria, but not in mitochondria, can be added. The result from testing a penicillin resistant E. coli (strain 2 in Table 1) is shown in Table 3 below. More specifically, the Minimal Inhibitory Concentration (MIC, g/ml) of AZT against E. coli strain no 2 in the presence of varying concentrations of uridine and respectively 2, 1 and 0.5 g/ml of trimethoprim is provided.

(50) TABLE-US-00003 TABLE 3 Trimethoprim Uridine concentration (g/ml) concentration (g/ml) 0 1 10 100 200 0 >20 25 >10 20 >10 20 >2 10 >2 10 2 >0.006 0.012 0.006 0.006 >0.012 0.025 >0.006 0.012 1 >0.012 0.025 0.006 0.006 >0.025 0.05 >0.012 0.025 0.5 >0.012 0.025 0.006 >0.05 0.1 >0.025 0.05 >0.012 0.025

(51) MIC of trimethoprim only against E. coli strain no 2 was established to be >816 g/ml

(52) A surprisingly strong synergistic effect against E. coli was observed for this combination according to the invention, and the presence of uridine will also reduce toxicity in patients.

Example 4

Synergistic Effect of AZT Combined with Uridine and Trimethoprim

(53) A further example is given in Table 4 below, which shows the synergistic effects against E. cloacae (strain 6 in Table 1) of combinations of AZT with trimethoprim and uridine. More specifically, the Minimal Inhibitory Concentration (MIC, g/ml) of AZT against E. cloacae in the presence of varying concentrations of uridine and respectively 2 and 1 g/ml of trimethoprim is provided.

(54) TABLE-US-00004 TABLE 4 Trimethoprim Uridine concentration (g/ml) concentration (g/ml) 0 10 100 200 0 >10 20 >5 10 >2 5 >2 5 2 >0 0.05 >0 0.05 >0 0.05 nt 1 >0 0.05 >0 0.05 >0 0.05 nt

(55) MIC of trimethoprim only against E. cloacae strain no 6 was established to be >1632 g/ml

Example 5

Further Bacteria Tested for AZT Combined with Uridine and Trimethoprim

(56) Table 5 below shows further bacterial strains tested for sensitivity to AZT with trimethoprim and uridine when used in a fixed ratio of concentrations.

(57) TABLE-US-00005 TABLE 5 Pathogen Denotation Properties Serratia marescens EN0454 Micrococcus luteus EN0455 Proteus mirabilis EN0457 Escherichia coli B EN0458 Pseudomonas aeruginosa EN0459 Pseudomonas aeruginosa EN0460 Enterobacter aerogenes EN0461 Klebsiella pneumoniae EN0463 Staphylococcus epidermidis EN0456 Citrobacter sp. EN0465 Salmonella typhimurium EN0466 Acinetobacter pittii or nosocomialis EN0180 Klebsiella oxytoca EN0392 Pseudomonas aeruginosa EN0422 Proteus mirabilis ST19 EN0445 Escherichia coli EN001 WT Escherichia coli EN002 DtoIC (isogenic to ATCC 25922) Escherichia coli EN003 D22 (lps mutant, hypersensitive) Pseudomonas aeruginosa EN004 WT Pseudomonas aeruginosa EN005 Efflux-defective (isogenic to PAO1) Klebsiella pneumoniae EN006 WT Acinetobacter baumannii EN007 WT Staphylococcus aureus EN008 Gram-positive control Klebsiella pneumoniae EN0010 WT Klebsiella pneumoniae EN0011 Efflux mutant Escherichia coli EN0012 WT Escherichia coli EN0013 DtoIC delition efflux mutant Pseudomonas aeruginosa EN0014 WT Pseudomonas aeruginosa EN0015 Efflux mutant Acinetobacter baumannii EN0016 WT Acinetobacter baumannii EN0017 Efflux mutant

Example 6

Strong Antibacterial Effects in Further Bacteria Tested

(58) The antibacterial effect (MIC, g/ml) of the combination of AZT, trimethoprim and uridine on the broad range of Gram-negative and Gram-positive bacterial pathogens and on selected mutant strains of Table 5 is shown in Table 6 below. Efflux mutations in K. pneumoniae, P. aeruginosa and A. baumanii were found to increase the sensitivity.

(59) TABLE-US-00006 TABLE 6 Substance concentration needed for total inhibition of pathogen growth (g/ml) Bacterial pathogen AZT Trimethoprim Uridine S. marescens EN0454 1 1 250 M. luteus EN0455 2 2 500 P. mirabilis EN0457 2 2 500 E. coli B EN0458 <0.008 <0.008 <2 P. aeruginosa EN0459 >4 >4 >1000 P. aeruginosa EN0460 >4 >4 >1000 E. aerogenes EN0461 0.25 0.25 60 K. pneumoniae EN0463 0.06 0.06 16 S. epidermidis EN0456 0.06 0.06 16 Citrobacter sp. EN0465 >4 >4 >1000 S. typhimurium EN0466 0.06 0.06 16 A. pittii or A. nosocomialis EN0180 >4 >4 >1000 K. oxytoca EN0392 0.06 0.06 16 P. aeruginosa EN0422 >4 >4 >1000 P. mirabilis ST19 EN0445 >4 >4 >1000 E. coli EN001 0.06 0.06 16 E. coli EN002 0.016 0.016 4 E. coli EN003 <0.008 <0.008 <2 P. aeruginosa EN004 >4 >4 >1000 P. aeruginosa EN005 1 1 250 K. pneumoniae EN006 0.06 0.06 16 A. baumannii EN007 >4 >4 >1000 S. aureus EN008 1 1 250 K. pneumoniae EN010 0.5 0.5 125 K. pneumoniae EN011 0.125 0.125 30 E. coli EN012 <0.008 <0.008 <2 E. coli EN013 <0.008 <0.008 <2 P. aeruginosa EN014 >4 >4 >1000 P. aeruginosa EN015 2 2 500 A. baumannii EN016 >4 >4 >1000 A. baumannii EN017 1 1 250

(60) The doses of AZT, uridine and trimethoprim can be given to patients to give steady state peak plasma levels of 0.1-10 g/ml of AZT, 0.5-5 g/ml of trimethoprim and 20-500 g/ml of uridine.

Example 7

FdUrd as the Antibacterial Nucleoside Analogue Shows Synergistic Effect Against E. cloacae and S. aureus when Combined with Uridine and Trimethoprim

(61) Table 7A below shows the Minimal Inhibitory concentration (MIC, g/ml) of FdUrd against E. cloacae strain 6 in the presence of varying concentrations of uridine and 2 and 1 g/ml of trimethoprim, respectively.

(62) TABLE-US-00007 TABLE 7A Trimethoprim MIC FdUrd (g/ml) (g/ml) Uridine 0 (g/ml) Uridine 10 (g/ml) Trimethoprim >4 8 N/A only 2 >1 2.5 >0.05 0.1 1 >2.5 >2.5 0 >30 >30

(63) Table 7B below shows the minimal Inhibitory concentration (MIC, g/ml) of FdUrd applied in combination with varying concentrations of trimethoprim against S.aureus strain 13 (MRSA) without and in the presence of uridine.

(64) TABLE-US-00008 TABLE 7B MIC FdUrd (g/ml) Trimeth- Uridine Trimethoprim oprim Trimethoprim Trimethoprim (g/ml) 0.25 (g/ml) 0.5 (g/ml) 1 (g/ml) 2 (g/ml) 0 >5 10 >1 5 >1 5 >1 5 10 >1 5 >1 5 >1 5 >0.1 1 100 >1 5 >1 5 >0.05 0.1 >0.05 0.1

(65) The addition of uridine will decrease toxicity of FdUrd in patients but not decrease the antibacterial effect and trimethoprim gives a synergistic antibacterial effect. The steady state peak level of FdUrd can be 0.2-2 g/ml in patients of trimethoprim 0.5-5 g/ml and of uridine 20-500 g/ml.

Example 8

Potent Antibacterial Effects of FdUrd with Trimethoprim and Uridine Against Several Gram Negative and Gram Positive Bacteria

(66) Table 8 below shows the antibacterial effect (MIC, g/ml) of the combination of FdUrd, trimethoprim and uridine on the broad range of Gram-negative and Gram-positive bacterial pathogens and on selected mutant strains (Table 5).

(67) TABLE-US-00009 TABLE 8 Substance concentration needed for total inhibition of pathogen growth (g/ml) Bacterial pathogen FdUrd Trimethoprim Uridine S. marescens EN0454 >4 >8 >400 M. luteus EN0455 2 4 200 P. mirabilis EN0457 1 2 100 E. coli B EN0458 0.032 0.064 3 P. aeruginosa EN0459 >4 >8 >400 P. aeruginosa EN0460 >4 >8 >400 E. aerogenes EN0461 1 2 100 K. pneumoniae EN0463 0.25 0.5 25 S. epidermidis EN0456 <0.008 <0.016 <0.8 Citrobacter sp. EN0465 >4 >8 >400 S. typhimurium EN0466 0.125 0.25 13 A. pittii or A. nosocomialis EN0180 4 8 400 K. oxytoca EN0392 0.125 0.25 13 P. aeruginosa EN0422 >4 >8 >400 P. mirabilis ST19 EN0445 0.5 1 50 E. coli EN001 0.125 0.25 13 E. coli EN002 0.032 0.064 6 E. coli EN003 0.016 0.032 1.6 P. aeruginosa EN004 >4 >8 >400 P. aeruginosa EN005 0.25 0.5 25 K. pneumoniae EN006 0.5 1 50 A. baumannii EN007 >4 >8 >400 S. aureus EN008 <0.008 <0.016 <0.8 K. pneumoniae EN010 0.5 1 50 K. pneumoniae EN011 0.064 0.125 6 E. coli EN012 0.032 0.064 3 E. coli EN013 0.016 0.032 1.6 P. aeruginosa EN014 >4 >8 >400 P. aeruginosa EN015 1 2 100 A. baumannii EN016 4 8 400 A. baumannii EN017 0.5 1 50

Example 9

Didanosine as the Antibacterial Nucleoside Analogue Shows Synergistic Effect Against E. coli when Combined with Uridine and Trimethoprim

(68) Table 9A below shows a potent effect against E. coli when combining didanosine and trimethoprim and uridine. Steady state peak plasma levels of didanosine can be 1-5 g/ml in patients, of trimethoprim 0.5-5 g/ml and of uridine 20-500 g/ml. More specifically, Table 9A provides the Minimal inhibitory concentration (MIC, g/ml) of didanosine against E. coli strain 2 in the presence of varying concentrations of uridine and 2 and 1 g/ml of trimethoprim, respectively.

(69) TABLE-US-00010 TABLE 9A MIC Didanosine (g/ml) Trimethoprim Uridine 0 Uridine 10 Uridine 100 (g/ml) (g/ml) (g/ml) (g/ml) Trimethoprim >8 16 N/A N/A only 2 >2.5 >2.5 >0.1 0.05 1 >2.5 >2.5 >0.1 0.05 0 >30 >30 >30

(70) Table 9B shows the inhibition of E. cloacae by combining didanosine, trimethoprim and uridine. More specifically, the Minimal Inhibitory Concentration (MIC, g/ml) of didanosine against E. cloacae strain 6 in the presence of varying concentrations of uridine and 2 and 1 g/ml of trimethoprim, respectively, is provided.

(71) TABLE-US-00011 TABLE 9B Trimethoprim MIC Didanosine (g/ml) (g/ml) Uridine 0 (g/ml) Uridine 10 (g/ml) 2 >2.5 >2.5 1 >1 2.5 >0.1 0.5 0 >30 >30

(72) MIC of trimethoprim only against E. cloacae strain 6 was established to be >1632 g/ml.

Example 10

Potent Antibacterial Effects of Didanosine with Trimethoprim and Uridine Against Several Gram-Negative and Gram-Positive Bacteria

(73) Table 10 shows the antibacterial effect (MIC, g/ml) of the combination of didanosine, trimethoprim and uridine on a broad range of Gram-negative and Gram-positive bacterial pathogens and on selected mutant strains (Table 5).

(74) TABLE-US-00012 Substance concentration needed for total inhibition of pathogen growth (g/ml) Bacterial pathogen Didanosine Trimethoprim Uridine S. marescens EN0454 >4 >8 >400 M. luteus EN0455 2 4 200 P. mirabilis EN0457 2 4 200 E. coli B EN0458 0.032 0.064 3 P. aeruginosa EN0459 >4 >8 >400 P. aeruginosa EN0460 >4 >8 >400 E. aerogenes EN0461 0.5 1 50 K. pneumoniae EN0463 0.25 0.5 25 S. epidermidis EN0456 0.064 0.125 6 Citrobacter sp. EN0465 >4 >8 >400 S. typhimurium EN0466 0.25 0.5 25 A. pittii or A. nosocomialis >4 >8 >400 EN0180 K. oxytoca EN0392 0.25 0.5 25 P. aeruginosa EN0422 >4 >8 >400 P. mirabilis ST19 EN0445 1 2 100 E. coli EN001 0.125 0.25 13 E. coli EN002 0.032 0.064 6 E. coli EN003 0.032 0.064 6 P. aeruginosa EN004 >4 >8 >400 P. aeruginosa EN005 0.5 1 50 K. pneumoniae EN006 0.5 1 50 A. baumannii EN007 >4 >8 >400 S. aureus EN008 0.5 1 50 K. pneumoniae EN010 0.5 1 50 K. pneumoniae EN011 0.125 0.25 13 E. coli EN012 0.064 0.125 6 E. coli EN013 0.016 0.032 1.6 P. aeruginosa EN014 >4 >8 >400 P. aeruginosa EN015 1 2 100 A. baumannii EN016 >4 >8 >400 A. baumannii EN017 1 2 100

Example 11

Influence of Uridine on the Effect of BrdUrd and Trimethoprim

(75) Table 11 below shows the MIC (g/ml) of BrdU applied in combination with 2 g/ml of trimethoprim without and with the presence of uridine against isolates of E. coli strain 2 and S. aureus strain 13.

(76) TABLE-US-00013 TABLE 11 MIC BrdUrd (g/ml) Pathogen Uridine (g/ml) E. coli strain 2 S. aureus strain 13 0 >16 32 >4 8 10 >8 16 2 100 >4 8 2

(77) MIC (g/ml) of trimethoprim only were established to be respectively >1632 g/ml (E. coli strain 2) and <32 (S. aureus strain 13).

Example 12

Influence of Thymidine on Antibacterial Effects

(78) The presence of thymidine in various tissues and in plasma under various disease conditions has to be taken into account when dosing AZT with trimethoprim and uridine, and similar combinations.

(79) Table 12 A and B shows the influence of thymidine on inhibition of E. coli and E. cloacae by AZT with trimethoprim and uridine at high thymidine concentrations Plasma and tissue concentrations of thymidine in humans have been reported to be in the range of 0.1 to 20 uM (0.025-5 g/ml).

(80) More specifically, Table 12 shows the Minimal Inhibitory Concentration (MIC, g/ml) of AZT against: (A) E. coli strain 2 and (B) E. cloacae strain 6 in the presence of respectively thymidine only, thymidine and 100 g/ml uridine as well as thymidine, 100 g/ml uridine and 1 g/ml trimethoprim.

(81) TABLE-US-00014 TABLE 12 MIC AZT (g/ml) Thymidine Uridine (U) and Trimethoprim (T) (g/ml) (M) 0 (U) 100 (U) 100 (U) and 1 (T) A 30 >30 >30 >0.5 10 >30 >30 >0.1 0.5 0 >20 30 >5 10 >0.05 0.01 B 30 >5 10 >10 >0.1 10 >5 10 >5 10 >0.05 0.1 0 >5 10 >2.5 5 0.001

Example 13

Enhanced Antibacterial Effect of AZT Plus Trimethoprim by Addition of Uridine in the Presence of Thymidine

(82) Table 13 below shows MIC values (ug/ml) of respectively AZT, trimethoprim and AZT+ trimethoprim mixed in propotions of 1:1 in the presence of 5 uM thymidine and added 0, 10, and 100 ug/ml of uridine against A) E. coli strain 2 and B) E. cloacae strain 6.

(83) TABLE-US-00015 TABLE 13 MIC (g/ml) Uridine AZT + (g/ml) AZT Trimethoprim Trimethoprim (1:1) A 0 >30 >5 10 >1.2 2.5 10 >25 30 >2.5 5 >0.3 0.6 100 >10 20 >2.5 5 >0.15 0.3 B 0 >5 10 <20 >0.04 0.075 10 >2.5 5.sup. <20 0.04 100 >2.5 5.sup. <20 0.04

Example 14

Enhanced Effect of AZT Plus Trimethoprim by Addition of Uridine in the Presence of Thymidine

(84) Table 14 shows the effect of uridine alone and uridine in the presence of 10 M thymidine on MIC (g/ml) of AZT/Trimethoprim combination mixed in proportions of 1:1 against A) E. coli strain 2; B) E. coli strain 6; and C) K. pneumoniae strain 8.

(85) TABLE-US-00016 TABLE 14 MIC AZT/Trimethoprim (1:1) (g/ml) Uridine Thymidine M (g/ml) 0 10 A 0 >0.3 0.6 >0.6 1.2 10 >0.3 0.6 >0.6 1.2 100 >0.15 0.3 >0.3 0.6 B 0 >0.04 0.08 >0.15 0.3 10 >0.04 0.08 >0.15 0.3 100 >0.02 0.04 >0.08 0.15 C 0 >0.08 0.15 >0.15 0.3 10 >0.08 0.15 >0.08 0.15 100 >0.04 0.08 >0.08 0.15

Example 15

Enhanced Effect of Uridine Alone and Uridine in the Presence of Thymidine

(86) The effect is shown of uridine alone and uridine in the presence of 10 M thymidine on MIC (g/ml) on an FdUrd/Trimethoprim combination mixed in proportions of 1:2 against A) E. coli strain 2; B) E. coli strain 6; C) K. pneumoniae strain 8, and D) S. aureus strain 13.

(87) TABLE-US-00017 TABLE 15 MIC FdUrd/Trimethoprim (1:2) (g/ml) Uridine Thymidine M (g/ml) 0 10 A 0 >10 20/20 40 >10 20/20 40 10 >10 20/20 40 >20/>40 100 >2.5 5/5 10 >20/>40 B 0 >0.3 0.6/0.6 1.2 >0.3 0.6/0.6 1.2 10 >0.3 0.6/0.6 1.2 >0.3 0.6/0.6 1.2 100 >0.3 0.6/0.6 1.2 >0.3 0.6/0.6 1.2 C 0 >1.2 2.5/2.5 5.sup. 2.5 5/>5 10 10 >0.6 1.2/>1.2 2.5 >0.6 1.2/>1.2 2.5 100 >0.6 1.2/>1.2 2.5 >0.6 1.2/>1.2 2.5 D FdUrd/Trimethoprim (1:2) Uridine Thymidine M (g/ml) 0 10 0 >5 10/10 20 10 20/>20 40 10 >2.5 5/>5 10 10 20/>20 40 100 >0.6 1.2/>1.2 2.5 >5 10/10 20.sup.

Example 16

Effect of Thymidine on the Inhibition by the Combination AZT with Trimethoprim and Uridine on a Panel of Gram-Negative and Gram-Positive Bacteria

(88) Table 16 below shows the antibacterial effect (MIC, g/ml) of the combination of AZT, trimethoprim and uridine only in comparison to the effect of the same combination mixed with thymidine on a broad range of Gram-negative and Gram-positive bacterial pathogens (described in Table 5).

(89) TABLE-US-00018 Substance concentration needed for total inhibition of pathogen growth g/ml Combination 1 Combination 2 Bacterial pathogen AZT Trimethoprim Uridine AZT Trimethoprim Uridine Thymidine (M) S. marescens EN0454 >4 >4 >400 >4 >4 >400 >120 M. luteus EN0455 2 2 200 >4 >4 >400 >120 P. mirabilis EN0457 2 2 200 >4 >4 >400 >120 E. coli B EN0458 <0.008 <0.008 <0.8 <0.008 <0.008 <0.8 <0.2 P. aeruginosa EN0459 >4 >4 >400 >4 >4 >400 >120 P. aeruginosa EN0460 >4 >4 >400 >4 >4 >400 >120 E. aerogenes EN0461 0.25 0.25 25 2 2 200 60 K. pneumoniae EN0463 0.125 0.125 12.5 0.25 0.25 25 8 S. epidermidis EN0456 0.064 0.064 66.4 2 2 200 60 Citrobacter sp. EN0465 >4 >4 >400 >4 >4 >400 >120 S. typhimurium EN0466 0.064 0.064 6.4 0.5 0.5 50 15 A. pittii or >4 >4 >400 >4 >4 >400 >120 A. nosocomialis EN0180 K. oxytoca EN0392 0.064 0.064 6.4 0.5 0.5 50 15 P. aeruginosa EN0422 >4 >8 >400 >4 >4 >400 >120 P. mirabilis ST19 EN0445 1 1 100 >4 >4 >400 >120 E. coli EN001 0.064 0.064 6.4 1 1 100 30 E. coli EN002 0.016 0.016 1.6 0.125 0.125 12.5 4 E. coli EN003 <0.008 <0.008 <0.8 <0.008 <0.008 <0.8 <0.2 P. aeruginosa EN004 >4 >4 >400 >4 >4 >400 >120 P. aeruginosa EN005 0.5 0.5 50 1 1 100 30 K. pneumoniae EN006 0.125 0.125 12.5 0.5 0.5 50 15 A. baumannii EN007 >4 >4 >400 >4 >4 >400 >120 S. aureus EN008 1 1 100 >4 >4 >400 >120 K. pneumoniae EN010 0.25 0.25 25 1 1 100 30 K. pneumoniae EN011 0.032 0.032 3.2 0.25 0.25 25 8 E. coli EN012 <0.008 <0.008 <0.8 <0.008 <0.008 <0.8 <0.2 E. coli EN013 <0.008 <0.008 <0.8 <0.008 <0.008 <0.8 <0.2 P. aeruginosa EN014 >4 >4 >400 >4 >4 >400 >120 P. aeruginosa EN015 2 2 200 2 2 200 60 A. baumannii EN016 >4 >4 >400 >4 >4 >400 >120 A. baumannii EN017 1 1 100 1 1 100 30

Example 17

Effect of AZT Combined with Trimethoprim and Uridine on Multiresistant Bacteria

(90) Table 17A shows the E. coli and K. pneumoniae strains used and their genotypes and resistance patterns. The resistance pattern means that the strains are clinically resistant to the antibiotics listed.

(91) TABLE-US-00019 TABLE 17 A Pathogen Denotation Genotype Resistance pattern E. coli EN0001 ATCC 25922, WT Wild type, control strain E. coli EN0134 AmpC (CMY-4) ESC, SXT E. coli EN0135 ESBL (SHV-2a) ESC (I), GEN, TET E. coli EN0136 ESBL (CTX-M-2) ESC, GEN, AMK, SXT E. coli EN0137 ESBL (CTX-M-15) ESC, CIP, GEN, SXT E. coli EN0138 ESBL (CTX-M-9) ESC, CIP, SXT, TET, CML E. coli EN0139 ESBL (CTX-M-15) ESC, CIP, AMK (I), SXT, TET E. coli EN0235 MG1655 gyrA S83L D87N CIP K. pneumoniae EN0006 ATCC 13883 WT Wild type, control strain K. pneumoniae EN0140 AmpC (DHA-1); OmpK36 loss ESC, CARBA, CIP, GEN + AMK, SXT, CML K. pneumoniae EN0233 ST (CC/CG) = 11(258) K. pneumoniae EN0142 CHDL (OXA-48); ESBL (CTX-M-15) ESC, ERT, CIP, GEN + TOB, SXT K. pneumoniae EN0143 ESBL (SHV-12) ESC, CIP, AMK (I), TOB, SXT K. pneumoniae EN0144 MBL (VIM-1); ESBL (SHV-5); AmpC ESC, CARBA, CIP, GEN, AMK, SXT, (CMY) TET K. pneumoniae EN0145 KPC (KPC-2) ESC, CARBA, CIP, AMK (I) K. pneumoniae EN0244 colistinR K. pneumoniae EN0245 colistinR K. pneumoniae EN0249 colistinR K. pneumoniae EN0262 ESBL (+) > (NP.+, MBL, KPC+, CTX-M, TEM+, OXA-1+) K. pneumoniae EN0401 Carbapenemase + ESBL K. pneumoniae EN0436 colistinR K. pneumoniae EN0437 DHA(+), CTX-M-1 colistinR P. aeruginosa EN0004 WT Wild type, control strain A. baumannii EN0007 WT Wild type, control strain

(92) The combination of AZT with trimethoprim and uridine was tested against E. coli and K. pneumoniae with resistance to several antibiotics. The combination treatment has surprisingly potent effects against these resistant strains, including strains resistant to trimethoprim-sulfamethoxazole.

(93) Table 17B shows the antibacterial effect (MIC, g/ml) of the combination of AZT, trimethoprim and uridine against a range of E. coli and K. pneumoniae strains with resistance to commercial antibiotics.

(94) TABLE-US-00020 Substance concentration needed for total inhibition of pathogen growth (g/ml) Bacterial pathogen AZT Trimethoprim Uridine E. coli EN0001 0.125 0.125 13 E. coli EN0134 1 1 100 E. coli EN0135 0.125 0.125 13 E. coli EN0136 0.125 0.125 13 E. coli EN0137 1 1 100 E. coli EN0138 0.016 0.016 1.6 E. coli EN0139 1 1 100 E. coli EN0235 <0.008 <0.008 <0.8 K. pneumoniae EN0006 0.25 0.25 25 K. pneumoniae EN0140 1 1 100 K. pneumoniae EN0233 0.25 0.25 25 K. pneumoniae EN0142 0.125 0.125 13 K. pneumoniae EN0143 >4 >4 >400 K. pneumoniae EN0144 1 1 100 K. pneumoniae EN0145 2 2 200 K. pneumoniae EN0244 0.5 0.5 50 K. pneumoniae EN0245 1 1 100 K. pneumoniae EN0249 1 1 100 K. pneumoniae EN0262 0.5 0.5 50 K. pneumoniae EN0401 >4 >8 >400 K. pneumoniae EN0436 0.125 0.125 13 K. pneumoniae EN0437 >4 >4 >400 P. aeruginosa EN0004 >4 >4 >400 A. baumannii EN0007 >4 >4 >400

Example 18

MIC Values for One of the Least Sensitive Strains of K. Pneumoniae

(95) Table 18 shows a comparison of MIC values for one of the least sensitive strains, K. pneumoniae EB0262. It is evident that the combination according to the invention is more potent than the 17 antibiotics listed, and that the inhibitory concentration can be used in patients.

(96) Table 18 shows the MIC values of AZT, trimethoprim and uridine combination against K.pneumoniae EN0262 (resistance pattern ESBL (+).fwdarw.(NP.+, MBL, KPC+, CTX-M, TEM+, OXA-1+) in comparison to MIC values of several commercial antibiotics.

(97) TABLE-US-00021 TABLE 18 Antibiotic MIC (g/ml) AZT; Trimethoprim; Uridine combination 0.5; 0.5; 50 AMP, ampicillin >128 AMC, amoxicillin- claculanic acid >64 PIP, piperacillin >256 TZP, piperacillin-tazobactam 64 CTX, cefotaxime >32 CAZ, ceftazidime >32 FEP, perfloxacin >16 ATM, aztreonam >16 IPM, imipenem >4 MEM, meropenem >8 GEN, gentamicin 1 AMK, amikacin 8 CIP, ciprofloxacin >2 SXT, trimethoprim-sulfamethoxazole 4 TET, tetracycline 4 TGC, tigecycline 2 CST, colistin 1 MIC data for commercial antibiotics against K. pneumoniae EN0262 (resistance pattern ESBL (+) > (NP.+, MBL, KPC+, CTX-M, TEM+, OXA-1+) were provided by IMBIM, BMC, Uppsala University.

Example 19

Effects of the Combination FdUrd, Trimethoprim and Uridine on Bacteria with Resistance to Commercial Antibiotics

(98) Table 19 shows the antibacterial effect (MIC, g/ml) of the combination of FdUrd, trimethoprim and uridine against a range of E. coli and K. pneumoniae strains with resistance to commercial antibiotics.

(99) TABLE-US-00022 Substance concentration needed for total inhibition of pathogen growth (g/ml) Bacterial pathogen FdUrd Trimethoprim Uridine E. coli EN0001 0.25 0.5 25 E. coli EN0134 >4 >8 >400 E. coli EN0135 0.25 0.5 25 E. coli EN0136 1 2 100 E. coli EN0137 >4 >8 >400 E. coli EN0138 0.25 0.5 25 E. coli EN0139 0.5 1 50 E. coli EN0235 0.06 0.125 6 K. pneumoniae EN0006 0.5 1 50 K. pneumoniae EN0140 >4 >8 >400 K. pneumoniae EN0233 0.5 1 50 K. pneumoniae EN0142 0.125 0.125 13 K. pneumoniae EN0143 >4 >8 >400 K. pneumoniae EN0144 >4 >8 >400 K. pneumoniae EN0145 >4 >8 >400 K. pneumoniae EN0244 >4 >8 >400 K. pneumoniae EN0245 >4 >8 >400 K. pneumoniae EN0249 >4 >8 >400 K. pneumoniae EN0262 2 4 200 K. pneumoniae EN0401 >4 >8 >400 K. pneumoniae EN0436 0.25 0.5 25 K. pneumoniae EN0437 >4 >8 >400 P. aeruginosa EN0004 >4 >8 >400 A. baumannii EN0007 >4 >8 >400

Example 20

Effects of the Combination Didanosine, Trimethoprim and Uridine on Bacteria with Resistance to Commercial Antibiotics

(100) Table 20 shows the antibacterial effect (MIC, g/ml) of the combination of didanosine, trimethoprim and uridine against a range of E. coli and K. pneumoniae strains with resistance to commercial antibiotics.

(101) TABLE-US-00023 Substance concentration needed for total inhibition of pathogen growth (g/ml) Bacterial pathogen Didanosine Trimethoprim Uridine E. coli EN0001 0.25 0.5 25 E. coli EN0134 >4 >8 >400 E. coli EN0135 0.25 0.5 25 E. coli EN0136 >4 >8 >400 E. coli EN0137 >4 >8 >400 E. coli EN0138 0.25 0.5 25 E. coli EN0139 0.5 1 50 E. coli EN0235 0.125 0.25 13 K. pneumoniae EN0006 0.5 1 50 K. pneumoniae EN0140 >4 >8 >400 K. pneumoniae EN0233 1 2 100 K. pneumoniae EN0142 >4 >8 >400 K. pneumoniae EN0143 >4 >8 >400 K. pneumoniae EN0144 >4 >8 >400 K. pneumoniae EN0145 >4 >8 >400 K. pneumoniae EN0244 >4 >8 >400 K. pneumoniae EN0245 >4 >8 >400 K. pneumoniae EN0249 >4 >8 >400 K. pneumoniae EN0262 2 4 200 K. pneumoniae EN0401 >4 >8 >400 K. pneumoniae EN0436 0.25 0.5 25 K. pneumoniae EN0437 >4 >8 >400 P. aeruginosa EN0004 >4 >8 >400 A. baumannii EN0007 >4 >8 >400

Example 21

MIC Values of FdUrd, Trimethoprim and Uridine and Respectively Didanosine, Trimethoprim and Uridine Combinations Against K. pneumoniae ENO262 (Resistance Pattern ESBL (+)(NP.+, MBL, KPC+, CTX-M, TEM+, OXA-1+) in Comparison to MIC of Several Commercial Antibiotics

(102) Table 21 shows the MIC values from Table 19 and 20 for FdUrd, trimethoprim and uridine and respectively didanosine, trimethoprim and uridine combinations against K. pneumoniae ENO262 (resistance pattern ESBL (+).fwdarw.(NP.+, MBL, KPC+, CTX-M, TEM+, OXA-1+) in comparison to MIC values of several commercial antibiotics.

(103) TABLE-US-00024 Antibiotic MIC (g/ml) FdUrd; Trimethoprim; Uridine 2; 4; 200 combination Didanosine; Trimethoprim; Uridine 2; 4; 200 combination AMP, ampicillin >128 AMC, amoxicillin-clavulanic acid >64 PIP, piperacillin >256 TZP, piperacillin-tazobactam 64 CTX, cefotaxime >32 CAZ, ceftazidime >32 FEP, cefepime >16 ATM, aztreonam >16 IPM, imipenem >4 MEM, meropenem >8 GEN, gentamicin 1 AMK, amikacin 8 CIP, ciprofloxacin >2 SXT, trimethoprim -sulfamethoxazole 4 TET, tetracycline 4 TGC, tigecycline 2 CST, colistin 1 MIC data for commercial antibiotics against K. pneumoniae EN0262 (resistance pattern ESBL (+) > (NP.+, MBL, KPC+, CTX-M, TEM+, OXA-1+) were provided by IMBIM, BMC, Uppsala University, Uppsala, Sweden.

Example 22

Effects of AZT Combined with Trimethoprim and Uridine and Effect of FdUrd Combined with Trimethoprim and Uridine Against Salmonella and Heliobacter

(104) The effects of AZT combined with trimethoprim and uridine are shown in Table 22A and the resistance patterns of the Salmonella strains in Table 22B. It is evident that the AZT+trimethoprim+uridine combination inhibits strains of Salmonella resistant also to trimethoprim-sulfa. Inhibition of Heliobacter is important in view of increasing resistance and its association to gastric cancer.

(105) TABLE-US-00025 TABLE 22A MIC (g/ml) AZT Trimethoprim Uridine FdUrd Trimethoprim Uridine Pathogen (g/ml) (g/ml) (g/ml) (g/ml) (g/ml) (g/ml) S. flexneri 1 1 100 >4 >8 >400 APV00906 S. flexneri 2 2 200 >4 >8 >400 APV00907 S. sonnei 0.03 0.03 3 >4 >8 >400 APV00908 Helicobacter 0.06 0.06 6 0.06 0.125 6 pylori APV00918 Helicobacter 0.125 0.125 13 0.06 0.125 6 pylori ATTC43504

(106) TABLE-US-00026 TABLE 22B Strain Antibiogram S. flexneri S. flexneri S. sonnei MIC (g/ml) APV00906 APV00907 APV00908 Amoxi/Clav S 4 | 16 S 2 Piper/tazo S 4 S 4 S 4 Cefotaxime S 64 S 1 S 1 Ceftazidime | 16 S 1 S 1 Cefrtriaxon / S S Cefepime S 2 S 1 S 1 Ertapenem S 0.5 S 0.5 S 0.5 Imipenem S 0.25 S 0.25 S 0.25 Meropenem S 0.25 S 0.25 S 0.25 Amikacin R* 16 R* 4 R* 4 Gentamicin R* 4 R* 1.sup. R* 1.sup. Ciproflaxin S 0.25 S 0.5 S 0.5 Tigeciclin S 0.5 S 0.5 S 0.5 Nitrofurantoin S 16 S 16 S 16 Trimet/sulfmeto R 320.sup. .sup.R 320 .sup.R 160

Example 23

In Vivo Effect of AZT Combined with Trimethoprim and Uridine

(107) This combination and AZT and trimethoprim separately were given to mice infected with E. coli. A murine peritonitis model was run, and the result is shown in Table 23B below. A potent inhibition was observed by the combination. A slight inhibition was obtained by dosing AZT and trimethoprim separately. Mice have been reported to have higher plasma concentrations of thymidine than humans. Table 23A shows the in vitro sensitivity of the E. coli strain used.

(108) TABLE-US-00027 TABLE 23A Plate MIC against E. coli ATCC 25922 A 0.06 g/ml AZT + 0.06 g/ml TMP with 50 g/ml uridine B 0.5 g/ml AZT C 0.5 g/ml TMP D 0.5 g/ml AZT + 0.5 g/ml TMP with 50 g/ml uridine and 2 g/ml thymidine E >2 g/ml AZT with 2 g/ml thymidine F 1 g/ml TMP with 2 g/ml thymidine

(109) TABLE-US-00028 TABLE 23B log diff log peritoneum mouse score at CFU/ml CFU/ml mean log vs start of Treatment no. sampling PF PF peritoneum treatment None 1 1 1.48E+05 5.17 5.30 2 1 1.45E+05 5.16 3 1 1.68E+05 5.23 4 1 4.50E+05 5.65 Trimethoprim 5 1 9.50E+03 3.98 5.34 0.04 30 mg/kg 6 1 1.60E+06 6.20 7 1 3.25E+05 5.51 8 1 4.75E+05 5.68 AZT 30 mg/kg 9 1 1.65E+04 4.22 5.68 0.38 10 1 4.25E+05 5.63 11 1 1.10E+07 7.04 12 1 6.75E+05 5.83 AZT + TMP + 13 1 8.75E+03 3.94 3.58 1.72 uridine 14 1 8.75E+03 3.94 30-30-50 15 1 5.50E+03 3.74 16 1 5.00E+02 2.70 AZT + TMP + 17 0 4.25E+03 3.63 3.45 1.85 uridine 18 0 4.25E+03 3.63 10-10-50 19 0 1.75E+03 3.24 20 0 2.00E+03 3.30 AZT + TMP + 21 1 1.13E+04 4.05 4.65 0.65 uridine 22 1 1.05E+04 4.02 3-3-50 23 1 3.25E+04 4.51 24 1 1.00E+06 6.00 AZT + TMP + 25 1 3.25E+05 5.51 5.79 0.49 uridine 26 2 5.00E+04 4.70 1-1-50 27 2 1.03E+07 7.01 28 2 8.25E+05 5.92 Ciprofloxacin 29 0 1.75E+03 3.24 2.91 2.39 14 mg/kg 30 0 2.50E+02 2.40 31 0 1.00E+03 3.00 32 0 1.00E+03 3.00 Vehicle 33 1 7.00E+07 7.85 7.36 2.06 34 1 1.58E+06 6.20 35 1 6.25E+07 7.80 36 1 7.75E+07 7.89 37 1 1.18E+07 7.07

Example 24

Triple Combinations with Sulfadiazine and Sulfamethoxazole Against E. coli

(110) Table 24A and 24B shows the MIC values (g/ml) of AZT against E. coli strain 2 in the presence of varying concentrations of uridine (g/ml) as well as trimethoprim and sulfamethoxazole mixed in ratios of 0.25:1.25 g/ml.

(111) TABLE-US-00029 TABLE 24 A MIC AZT (g/ml) Sulfadiazine 1.25 g/ml Uridine (g/ml) Trimethoprim 0.25 g/ml 0 >0.5 1 10 >0.5 1 100 >0.25 0.5

(112) TABLE-US-00030 Table 24 B MIC AZT (g/ml) Sulfamethoxazol 1.25 g/ml Uridine (g/ml) Trimethoprim 0.25 g/ml 0 >0.5 1 10 >0.05 1 100 0.05

Example 25

Synergistic Effect of AZT Combined with Uridine and Iclaprim Against S. aureus and E. coli

(113) TABLE-US-00031 TABLE 25A Isolate S. aureus no. 13 MIC Iclaprim (g/ml) Uridine 0 Uridine 100 Uridine 300 Uridine 500 Isolate (g/ml) (g/ml) (g/ml) (g/ml) S. aureus >16 >16 >0.06 0.12 >0.06 0.12 no. 13

(114) TABLE-US-00032 TABLE 25B Isolate S. aureus no. 13 MIC AZT (g/ml) Uridine 0 Uridine 100 Uridine 300 Uridine 500 Iclaprim (g/ml) (g/ml) (g/ml) (g/ml) 0 >32 >32 >32 >32 0.08 >2 4 >2 4 >0.03 0.06 >0.03 0.06 0.015 >2 4 >2 4 >0.008 0.015 >0.015 0.03 0.03 >2 4 >0.25 0.5 0.008 0.008

(115) TABLE-US-00033 TABLE 25C Isolate E. coli no. 2 MIC Iclaprim (g/ml) Uridine 0 Uridine 100 Uridine 300 Uridine 500 Isolate (g/ml) (g/ml) (g/ml) (g/ml) E. coli no. 2 >2 4 >2 4 >0.25 0.5 >0.25 0.5

(116) TABLE-US-00034 TABLE 25D Isolate E. coli no. 2 MIC AZT (g/ml) Uridine 0 Uridine 100 Uridine 300 Uridine 500 Iclaprim (g/ml) (g/ml) (g/ml) (g/ml) 0 >20 25 >20 25 >5 15 >2 10 0.1 >8 >2 4 >0.25 0.5 >0.25 0.5 0.2 >8 >1 2 >0.12 0.25 >0.25 0.5 0.3 >1 2 >0.25 0.5 >0.12 0.25 >0.25 0.5

Example 26

Synergistic Effect of FdUrd Combined with Uridine and Iclaprim Against S. aureus and E. coli

(117) TABLE-US-00035 TABLE 26A MIC FdUrd (g/ml) Uridine 0 Uridine 100 Uridine 300 Uridine 500 Iclaprim (g/ml) (g/ml) (g/ml) (g/ml) 0 >4 8 >2 4 >0.03 0.06 >0.03 0.06 0.08 >0.5 >0.25 0.5 >0.004 0.008 >0.015 0.03 0.015 >0.5 >0.25 0.5 >0.004 0.008 >0.004 0.008 0.03 >0.5 >0.25 0.5 >0.004 0.008 >0.004 0.008

(118) TABLE-US-00036 TABLE 26B MIC FdUrd (g/ml) Uridine 0 Uridine 100 Uridine 300 Uridine 500 Iclaprim (g/ml) (g/ml) (g/ml) (g/ml) 0 >16 >16 >16 >16 0.1 >8 >8 >1 2 >1 2 0.2 >8 >2 4 >0.5 1 >1 2 0.3 >1 2 >0.25 0.5 >0.06 0.12 >0.06 0.12

Example 27

Effect of Combinations of Zidovudine (AZT), Fluorodeoxyuridine (FdUrd), Trimethoprim (TMP), Iclaprim (ICP) and Uridine (U). MIC Values are in g/ml for AZT/FdUrd

(119) TABLE-US-00037 AZT + AZT + FdUrd + FdUrd + TMP + U ICP + U TMP + U ICP + U Staphylococcus 1 >4 0.5 <0.015 aureus Staphylococcus 0.06 nt <0.008 <0.008 epidermis Streptococcus nt nt 1-2 0.06 pneumoniae Micrococcus 2 nt 2 nt luteus Bacillus anthracis >4 <0.25 0.05 Bacillus cereus nt nt nt nt Acinetobacter 1, >4 nt 0.5; 4 nt baumannii Acinetobacter >4 nt 4 nt pittii Escherichia coli <0.01; 1 <0.02-0.12 0.016; >4 >0.1-1 Klebsiella 0.03; >4 0.2 0.06; >4 nt pneumoniae Klebsiella 0.06 nt 0.125 nt oxytoca Enterobacter 0.04 0.02 0.3 nt cloacae Enterobacter 0.25 nt 1 nt aerogenes Serratia 1; >4 nt >4 nt marcescens Citrobacter >4 nt >4 nt Proteus mirabilis 2; >4 nt 1 nt Pseudomonas 1; >4 nt 0.25; >4 nt aeruginosa Salmonella 0.06 0.016-0.06 0.25 0.5-1 typhimurium Shigella flexneri 1; 2 >4 >4 >4 Shigella sonnei 0.03 0.03 >4 2 Campylobacter >4 nt >4 nt jejuni Helicobacter 0.06; 0.1 >4 0.06 >4 pylori Yersinia pestis nt 2 nt 2 Haemophilus 0.125-4 0.25-4 0.25-2 0.25-2 influenza Clostridium >4 nt >4 nt difficile

Example 28

Effect of Combinations of Gemcitabine (GEM), Didanosine (DID), Trimethoprim (TMP), Iclaprim (ICP) and Uridine (U). MIC Values are in g/ml for GEM/DID

(120) TABLE-US-00038 GEM + GEM + DID + DID + TMP + U ICP + U TMP + U ICP + U Staphylococcus >0.08->0.015 nt 0.5 >8 aureus Staphylococcus nt nt 0.064 nt epidermis Streptococcus nt 0.06 nt nt pneumoniae Micrococcus nt nt 2 nt luteus Bacillus <0.25 <0.05 nt nt anthracis Bacillus cereus nt nt nt nt Acinetobacter nt nt 1->4 nt baumannii Acinetobacter nt nt >4 nt pittii Escherichia coli >4 nt 0.03->4 >8 Klebsiella nt nt 0.125->4 1-2 pneumoniae Klebsiella nt nt 0.25 nt oxytoca Enterobacter nt nt 0.5->2.5 0.015-0.03 cloacae Enterobacter nt nt 0.5 nt aerogenes Serratia nt nt >4 nt marcescens Citrobacter nt nt >4 nt Proteus mirabilis nt nt 1.2 nt Pseudomonas nt nt 0.5->4 nt aeruginosa Salmonella nt nt 0.25 nt typhimurium Shigella flexneri nt nt nt nt Shigella sonnei nt nt nt nt Campylobacter nt 4->4 nt nt jejuni Helicobacter nt nt nt nt pylori Yersinia pestis nt <2 nt nt Haemophilus nt nt nt nt influenza Clostridium nt >4 nt nt difficile

Example 29

Effect of Combinations of Nicavir (NIC), AZT Phosphonate (M2), Trimethoprim (TMP), Iclaprim (ICP) and Uridine (U). MIC Values are in g/ml for NIC/M2

(121) TABLE-US-00039 NIC + NIC + M2 + M2 + TMP + U ICP + U TMP + U ICP + U Staphylococcus >40 >16 0.1, >32 >8 aureus Staphylococcus nt nt nt nt epidermis Streptococcus nt 0.03-0.5 nt nt pneumoniae Micrococcus nt nt nt nt luteus Bacillus anthracis nt >4 nt nt Bacillus cereus nt nt nt nt Acinetobacter >40 nt >30 nt baumannii Acinetobacter nt nt nt nt pittii Escherichia coli 0.04-0.4 2.4 >20 >8 Klebsiella >4 nt >30 >8 pneumoniae Klebsiella oxytoca nt nt nt nt Enterobacter 0.04-0.4 nt <0.005 >8 cloacae Enterobacter nt nt nt nt aerogenes Serratia nt nt nt nt marcescens Citrobacter nt nt nt nt Proteus mirabilis nt nt nt nt Pseudomonas >40 nt >30 nt aeruginosa Salmonella nt nt nt nt typhimurium Shigella flexneri nt nt nt nt Shigella sonnei nt nt nt nt Campylobacter nt 4->4 nt nt jejuni Helicobacter nt nt nt nt pylori Yersinia pestis nt >2 nt nt Haemophilus nt nt nt nt influenza Clostridium nt >4 nt nt difficile

Example 30

Synergistic Effect of FdUrd Combined with Uridine and Iclaprim Against Strains of S. aureus

(122) MIC of FdUrd tested in the presence of 0.008; 0.015; and 0.03 g/ml iclaprim, respectively, and with rising concentrations of uridine (0, 100, 300 and 500 g/ml) against 7 different isolates of S.aureus. For some of the isolates, 0.004 or 0.06 Wml of iclaprim was used additionally.

(123) TABLE-US-00040 TABLE 30A Isolate S. aureus no. 11 MIC FdUrd (g/ml) Iclaprim Uridine 0 Uridine 100 Uridine 300 Uridine 500 (g/ml) (g/ml) (g/ml) (g/ml) (g/ml) 0 >8 16 >8 16 >0.06 0.12 >0.06 0.12 0.008 >4 >4 >0.008 0.015 >0.008 0.015 0.015 >4 >4 >0.004 0.008 >0.004 0.008 0.03 >4 >4 >0.004 0.008 >0.004 0.008

(124) TABLE-US-00041 TABLE 30B Isolate S. aureus no. 12 MIC FdUrd (g/ml) Iclaprim Uridine 0 Uridine 100 Uridine 300 Uridine 500 (g/ml) (g/ml) (g/ml) (g/ml) (g/ml) 0 >8 16 >2 4 >0.06 0.12 >0.03 0.06 0.008 >4 >2 4 >0.06 0.12 >0.03 0.06 0.015 >4 >2 4 >0.03 0.06 >0.015 0.03 0.03 >4 >2 4 >0.03 0.06 >0.015 0.03

(125) TABLE-US-00042 TABLE 30C Isolate S. aureus no. 13 MIC FdUrd (g/ml) Iclaprim Uridine 0 Uridine 100 Uridine 300 Uridine 500 (g/ml) (g/ml) (g/ml) (g/ml) (g/ml) 0 >4 8 >1 2 >0.03 0.6 >0.03 0.06 0.008 >4 >1 2 >0.015 0.03 >0.008 0.015 0.015 >4 >1 2 >0.008 0.015 >0.008 0.015 0.03 >4 >0.5 1 >0.008 0.015 >0.008 0.015

(126) TABLE-US-00043 TABLE 30D Isolate S. aureus V55 MIC FdUrd (g/ml) Iclaprim Uridine 0 Uridine 100 Uridine 300 Uridine 500 (g/ml) (g/ml) (g/ml) (g/ml) (g/ml) 0 >1 2 >0.5 1 >0.03 0.6 >0.015 0.03 0.004 >1 2 >0.06 0.12 >0.008 0.015 >0.008 0.015 0.008 >1 2 >0.06 0.12 >0.008 0.015 >0.008 0.015 0.015 >1 2 >0.06 0.12 >0.008 0.015 >0.008 0.015 0.03 >1 2 >0.06 0.12 >0.008 0.015 >0.008 0.015

(127) TABLE-US-00044 TABLE 30E Isolate S. aureus V86 MIC FdUrd (g/ml) Iclaprim Uridine 0 Uridine 100 Uridine 300 Uridine 500 (g/ml) (g/ml) (g/ml) (g/ml) (g/ml) 0 >4 8 >2 4 >0.03 0.06 >0.03 0.06 0.008 >4 >2 4 >0.015 0.03 >0.015 0.03 0.015 >4 >2 4 >0.015 0.03 >0.015 0.03 0.03 >4 >2 4 >0.015 0.03 >0.015 0.03 0.06 >4 >2 4 >0.008 0.015 >0.008 0.015

(128) TABLE-US-00045 TABLE 30F Isolate S. aureus V308 MIC FdUrd (g/ml) Iclaprim Uridine 0 Uridine 100 Uridine 300 Uridine 500 (g/ml) (g/ml) (g/ml) (g/ml) (g/ml) 0 >16 >16 >2 4 >1 2 0.008 >4 >4 >0.03 0.06 >0.03 0.06 0.015 >4 >4 >0.03 0.06 >0.03 0.06 0.03 >4 >2 4 >0.03 0.06 >0.03 0.06 0.06 >4 >2 4 >0.03 0.06 >0.03 0.06

(129) TABLE-US-00046 TABLE 30G Isolate S. aureus V321 MIC FdUrd (g/ml) Iclaprim Uridine 0 Uridine 100 Uridine 300 Uridine 500 (g/ml) (g/ml) (/ml) (g/ml) (g/ml) 0 >0.5 1 >0.25 0.5 >0.03 0.06 >0.03 0.06 0.008 >0.5 1 >0.25 0.5 >0.015 0.03 >0.008 0.015 0.015 >0.25 0.5 >0.25 0.5 >0.015 0.03 >0.004 0.008 0.03 >0.25 0.5 >0.25 0.5 >0.015 0.03 >0.004 0.008

(130) Resistance patterns for S. aureus isolates

(131) TABLE-US-00047 S. aureus no. 11 Wt S. aureus no. 12 MSSA/weak penicillin producer S. aureus no. 13 MRSA S. aureus V55 Mupirocin resistant, MIC >256 g/ml S. aureus V86 Mupirocin resistant, MIC >256 g/ml S. aureus V308 Mupirocin resistant, MIC >256 g/ml S. aureus V321 Mupirocin resistant, MIC >256 g/ml