Compounds and their use in the treatment of non-viral diseases and infections

Abstract

The present invention relates to compounds having a structure according to general formula I or II or III for use in the treatment of non-viral infections or a disease caused by a non-viral infection. The present invention relates to compounds having a structure according to general formula III and to pharmaceutical compositions comprising at least one of said compounds. The present invention further relates to a method of treating non-viral infections or a disease caused by a non-viral infection.

Description

[0001] COMPOUNDS AND THEIR USE IN THE TREATMENT OF NON- VIRAL

[0002] DISEASES AND INFECTIONS

[0003] The present invention relates to compounds having a structure according to general formula I or II or III for use in the treatment of non-viral infections or a disease caused by a non-viral infection. The present invention further relates to a method of treating non-viral infections or a disease caused by a non-viral infection. The present invention relates to compounds having a structure according to general formula III and to pharmaceutical compositions comprising at least one of said compounds.

[0004] BACKGROUND OF THE INVENTION

[0005] The spread of antibacterial resistance has been deemed one of the greatest challenges of mankind by the world's leaders at the 2017 G20 summit (G20 Leaders Declaration Shaping an interconnected world. (2017)). ‘No time to wait’ - in 2019 the World Health Organization (WHO) released an urgent warning over antimicrobial resistance, a global crisis that threatens a century of progress (Interagency Coordination Group on Antimicrobial Resistance: No time to wait: securing the future from drug-resistant infections, World Health Organisation (2019)). The challenges associated with antimicrobial resistance are complex and multifaceted, but it is not too late to overcome the threat by drug-resistant infections if action is taken now. The development of drug-resistant bacteria is mainly attributed to their extensive overuse and misuse leading to a continuous selection pressure on bacteria (Davies and Davies, 2010; CDC. Antibiotic Resistance Threats in the United States, 2019. Atlanta, GA: U.S. Department of Health and Human Services, CDC (2019)). Surprisingly, scientific data regarding the exact mode of actions and involved protein targets are lacking for several antibiotics in clinical use. This hinders the understanding of resistance formation and the identification of new antibiotics for development of next-generation drugs (Bandow et al., 2003; Ang etal., 2017).

[0006] One example of poorly understood mode of action is that of the commonly used antibiotic metronidazole. Metronidazole has been considered the standard treatment of several infectious diseases for more than 45 years - mainly attributed to the lack of alternatives - against several protozoal infections, anaerobic bacteria (e.g. Clostridioides difficile') and microaerophilic bacteria (e.g. Helicobacter pylori) (Lbfmark etal., 2010). However, although metronidazole is broadly applied in the treatment of H. pylori and C. difficile, its exact mode of action remains to be elucidated and alternatives with improved efficacy especially against metronidazole- resistant strains are urgently needed.

[0007] Helicobacter pylori infections are characterized by increasing antibiotic resistance affecting the efficacy of current treatment options (Vakil et al., 2007; Megraud el al., 2004). In Europe, H. pylori resistance rates for adults are 18 % for clarithromycin, 14 % for levofloxacin and as much as 35% for metronidazole and these resistances are the major cause for treatment failures (Ang etal., 2017). H. pylori is a gram-negative, microaerophilic bacterium and responsible for numerous gastrointestinal disorders including gastritis, peptic ulcer disease or even gastric cancer (Perez -Perez et al. , 2004; Megraud et al., 2004). Furthermore, colonization in the human stomach persists lifelong unless treated (Lehours et al., 2007). It is the first formally recognized bacterial carcinogen and a prominent human pathogen as 50 % of the global population are infected. In developing countries, prevalence of infection is up to 80%. Among the infected 15% will develop gastritis and 1% cancer (Ang et al., 2017). Hence, developing new or improved drugs, improving treatment options, and monitoring antibiotic resistance of H. pylori is vital for infection management in clinical practice (Ang et al., 2017; Boucher et al, 2009; Zagari et al, 2018).

[0008] Jamshidi et al. (2022) describe triazole / quinoline hybrid compounds that have antifungal activity, such as against S. cervisiae and C. albicans, as well as activity against aerobic bacteria, such as E. coli and Staphylococcus aureus. Li et al. (2019) describe indol-nitroimidazole conjugates that decrease the gene expression of methicillin-resistant Staphylococcus aureus. Foroumadi et al. (2004) describe the in vitro antituberculosis activity of two thiadiazole derivatives.

[0009] Thus, there is a great need for new compounds having antibacterial activity for the treatment of non-viral diseases and infections, in particular for the treatment of bacterial diseases caused by bacteria resistant to commonly used antibiotic agents.

[0010] It is an objective of the present invention to provide novel compounds and their use in the treatment of non-viral infections or a disease caused by a non-viral infection.

[0011] SUMMARY OF THE INVENTION According to the present invention this object is solved by providing a compound having a structure according to general formula I or II or III

[0012] (I) (II) (HI) wherein

[0013] R1is Ci-Ce alkyl or Ci-Ce haloalkyl,

[0014] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, wherein said Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl is optionally substituted with halogen, OR3or SR3, wherein R3is hydrogen or Ci-Ce alkyl, or a pharmaceutically acceptable salt thereof, under the proviso that for a compound of general formula II or III: when R1is methyl then R2is not ethyl or C1-C4 alkyl substituted with halogen, OH, OMe, NH2, NH(CH3)2or NH(CH2-CH3)2, when R1is CF3 then R2is not C1-C4 alkyl substituted with halogen, when R1is isopropyl then R2is not C1-C4 alkyl substituted with halogen; under the proviso that for a compound of general formula III: not both R1and R2are methyl, for use in the treatment of non-viral infections or a disease caused by a non-viral infection.

[0015] According to the present invention this object is solved by providing a method for the treatment of non-viral infections or a disease caused by a non-viral infection, comprising the step of administering to a subject in need thereof a compound having general formula I or II or III according to the present invention or a pharmaceutical composition of the present invention. According to the present invention this object is solved by providing a compound having a structure according to general formula III wherein

[0016] R1is Ci-Ce alkyl or Ci-Ce haloalkyl, and

[0017] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, wherein said Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl is optionally substituted with halogen, OR3or SR3, wherein R3is hydrogen or Ci-Ce alkyl, or a pharmaceutically acceptable salt thereof, under the proviso that not both R1and R2are methyl, when R1is methyl then R2is not ethyl or C1-C4 alkyl substituted with halogen, OH, OMe, NH2, or NH(CH3)2, when R1is CF3 then R2is not C1-C4 alkyl substituted with halogen, when R1is isopropyl then R2is not C1-C4 alkyl substituted with halogen.

[0018] According to the present invention this object is solved by providing a pharmaceutical composition comprising

[0019] (i) at least one compound having a structure according to general formula III of the present invention,

[0020] (ii) optionally, pharmaceutical excipient(s) and / or carrier.

[0021] According to the present invention this object is solved by providing a compound having a structure according to general formula III of the present invention for use in medicine.

[0022] DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION

[0023] Before the present invention is described in more detail below, it is to be understood that this invention is not limited to the particular methodology, protocols and reagents described herein as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. For the purpose of the present invention, all references cited herein are incorporated by reference in their entireties.

[0024] Concentrations, amounts, and other numerical data may be expressed or presented herein in a range format. It is to be understood that such a range format is used merely for convenience and brevity and thus should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. As an illustration, a numerical range of "1 to 20" should be interpreted to include not only the explicitly recited values of 1 to 20, but also include individual values and sub-ranges within the indicated range. Thus, included in this numerical range are individual values such as 1, 2, 3, 4, 5 ... . 17, 18, 19, 20 and sub-ranges such as from 2 to 10, 8 to 15, etc. This same principle applies to ranges reciting only one numerical value, such as "higher than 150 mg per day". Furthermore, such an interpretation should apply regardless of the breadth of the range or the characteristics being described.

[0025] As used herein and throughout the entire description, the term "alkyl" refers to a monoradical of a saturated straight or branched hydrocarbon. Preferably, the alkyl group comprises from 1 to 6 carbon atoms, i.e., 1, 2, 3, 4, 5 or 6 carbon atoms, more preferably 1 to 4 carbon atoms. In some embodiments, the alkyl group employed in the invention contains 1-6 carbon atoms (Cn 6 alkyl). In another embodiment, the alkyl group employed contains 1-5 carbon atoms (C1-5 - alkyl). In another embodiment, the alkyl group employed contains 1-4 carbon atoms (C alkyl). In another embodiment, the alkyl group employed contains 1-3 carbon atoms (C1-3 alkyl). In another embodiment, the alkyl group employed contains 1-2 carbon atoms (C1-2 alkyl). In another embodiment, the alkyl group employed is methyl. Examples of alkyl radicals include, but are not limited to, methyl, ethyl, propyl, iso-propyl, n-propyl, butyl, iso-butyl, n-butyl, tertbutyl, n-pentyl, iso-pentyl, sec-pentyl, neo-pentyl, 1,2-dimethyl-propyl, iso-amyl, n-hexyl, isohexyl, sec-hexyl, and the like, which may bear one or more substituents. Alkyl group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety. In some embodiments the alkyl chain is linear. In some embodiments the alkyl chain is branched. In some embodiments the alkyl chain is substituted. In some embodiments the alkyl chain is unsubstituted. In some embodiments the alkyl chain is linear and substituted or unsubstituted. In some embodiments the alkyl chain is branched and substituted or unsubstituted.

[0026] As used herein and throughout the entire description, the term "alkenyl" refers to a monoradical of a straight or branched hydrocarbon containing at least one double bond (-C=C-). Preferably, the alkenyl group comprises from 2 to 5 carbon atoms, i.e. 2, 3, 4, 5 or 6 carbon atoms, more preferably 1 to 4 carbon atoms. In some embodiments, the alkenyl group employed in the invention contains 2-6 carbon atoms (C2-6 alkenyl). In another embodiment, the alkenyl group employed contains 2-5 carbon atoms (C2-5 alkenyl). In another embodiment, the alkenyl group employed contains 2-4 carbon atoms (C2-4 alkenyl). In another embodiment, the alkenyl group employed contains 2-3 carbon atoms (C2-3 alkenyl). In another embodiment, the alkenyl group employed contains 2 carbon atoms (C2 alkenyl). Examples of alkenyl radicals include, but are not limited to, allyl, vinyl, dimethylallyl, 1-butenyl, 2-butenyl, 3-butenyl, 1- pentenyl, 2- pentenyl, 3-pentenyl, or 4-pentenyl, and the like, which may bear one or more substituents. Alkenyl group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety. In some embodiments the alkenyl chain is linear. In some embodiments the alkenyl chain is branched. In some embodiments the alkenyl chain is substituted. In some embodiments the alkenyl chain is unsubstituted. In some embodiments the alkenyl chain is linear and substituted or unsubstituted. In some embodiments the alkenyl chain is branched and substituted or unsubstituted.

[0027] As used herein and throughout the entire description, the term "alkynyl" refers to a monoradical of a straight or branched hydrocarbon containing at least one triple bond (-C=C-). Preferably, the alkynyl group comprises from 3 to 6 carbon atoms, i.e. 3, 4, 5 or 6 carbon atoms, more preferably 3 to 4 carbon atoms. In some embodiments, the alkynyl group employed in the invention contains 3-6 carbon atoms (C3-6 alkynyl). In another embodiment, the alkynyl group employed contains 3-5 carbon atoms (C3-5 alkynyl). In another embodiment, the alkynyl group employed contains 3-4 carbon atoms (C2-4 alkynyl). In another embodiment, the alkynyl group employed contains 4 carbon atoms (C4 alkynyl). In another embodiment, the alkynyl group employed contains 3 carbon atoms (C3 alkynyl). Examples of alkenyl radicals include, but are not limited to, propargyl, but-l-yne, 1-butynyl, 2-butynyl, 3-butynyl, 1-propynyl, or 2- propynyl, and the like, which may bear one or more substituents. Alkynyl group substituents include, but are not limited to, any of the substituents described herein, that result in the formation of a stable moiety. In some embodiments the alkynyl chain is linear. In some embodiments the alkynyl chain is branched. In some embodiments the alkynyl chain is substituted. In some embodiments the alkynyl chain is unsubstituted. In some embodiments the alkynyl chain is linear and substituted or unsubstituted. In some embodiments the alkynyl chain is branched and substituted or unsubstituted.

[0028] As used herein and throughout the entire description, the term “haloalkyl” refers to an alkyl group substituted by one halogen substituent up to per halo- substitution. The halogen substituent is preferably fluorine. The haloalkyl is preferably a perfluoroalkyl. In some embodiments, the haloalkyl group employed in the invention contains 1-6 carbon atoms (Ci-6 haloalkyl). In another embodiment, the haloalkyl group employed in the invention contains 1- 5 carbon atoms (C1-5 haloalkyl). In another embodiment, the haloalkyl group employed in the invention contains 1-4 carbon atoms (C1-4 haloalkyl). In another embodiment, the haloalkyl group employed in the invention contains 1-3 carbon atoms (C1-3 haloalkyl). In another embodiment, the haloalkyl group employed in the invention contains 1-2 carbon atoms (C1-2 haloalkyl). In another embodiment, the haloalkyl group employed in the invention contains 1- carbon atom (Ci haloalkyl). In another embodiment, the haloalkyl group employed in the invention is trifluorom ethyl. Exemplary fluoro-substituted C1-C2 alkyl includes -CFH2, -CF2H, -CF3, CH2CH2F, -CH2CHF2, -CHFCH3, -CHFCH3, -CF2CHF2. Perfluoro-substituted C1-C2 haloalkyl, for example include -CF3, and -CF2CF3. In some embodiments haloalkyl is Ci haloalkyl, in particular -CF3.

[0029] As used herein and throughout the entire description, the term “substituted” indicates that a group, such as an alkyl, an alkenyl, or an alkynyl, may be substituted with one or more substituents, such as substituted with one or more of halogen, OR3or SR3wherein R3is hydrogen or Ci-Ce alkyl. “Substituted” in reference to a group indicates that one or more hydrogen atoms attached to a member atom within the group is replaced with a substituent selected from the group of defined or suitable substituents. It should be understood that the term “substituted” includes the implicit provision that such substitution be in accordance with the permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound. When it is stated that a group may contain one or more substituents, one or more member atoms within the group may be substituted. In addition, a single member atom within the group may be substituted with more than one substituent as long as such substitution is in accordance with the permitted valence of the atom.

[0030] Nitroimidazole compounds for treating non-viral infections

[0031] As outlined above, the present invention provides a compound having general formula I or II or III for use in the treatment of non-viral infections or a disease caused by a non-viral infection.

[0032] A compound having general formula I or II or III

[0033] (I) (II) (HI) is a compound wherein

[0034] R1is Ci-Ce alkyl or Ci-Ce haloalkyl,

[0035] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, wherein for R2said Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl is optionally substituted with halogen, OR3or SR3, wherein R3is hydrogen or Ci-Ce alkyl, or a pharmaceutically acceptable salt thereof.

[0036] - Disclaimer

[0037] According to the invention, a compound for use according to the present invention is not a compound of general formula II or III wherein

[0038] R1is methyl and

[0039] R2is ethyl or C1-C4 alkyl substituted with halogen, OH, OMe, NH2, NH(CH3)2 or NH(CH2-CH3)2. According to the invention, a compound for use according to the present invention is not a compound of general formula II or III wherein

[0040] R1is CF3 and

[0041] R2is C1-C4 alkyl substituted with halogen.

[0042] According to the invention, a compound for use according to the present invention is not a compound of general formula II or III wherein

[0043] R1is isopropyl and

[0044] R2is C1-C4 alkyl substituted with halogen.

[0045] According to the invention, a compound for use according to the present invention is not a compound of general formula III wherein both R1and R2are methyl.

[0046] In one embodiment of a compound for use according to the present invention, R1is methyl and the compound has formula la or Ila or Illa

[0047] In one embodiment of a compound for use according to the present invention:

[0048] R2is Ci-Ce alkyl, preferably C1-C4 alkyl, more preferably methyl, ethyl, propyl, isopropyl, n- propyl, n-butyl, isobutyl, sec-butyl, tert-butyl, neopentyl, or n-hexyl.

[0049] In one embodiment of a compound for use according to the present invention:

[0050] R2is C2-C6 alkenyl, preferably C2-C5 alkenyl, more preferably allyl, vinyl, dimethylallyl, 1- butenyl, 2-butenyl, 3-butenyl, 1- pentenyl, 2-pentenyl, 3 -pentenyl, or 4-pentenyl.

[0051] In one embodiment of a compound for use according to the present invention: R2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, but-l-yne, 1- butynyl, 2-butynyl, 3-butynyl, 1-propynyl, or 2-propynyl.

[0052] In one embodiment of a compound for use according to the present invention:

[0053] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, which is substituted with halogen.

[0054] In one embodiment of a compound for use according to the present invention:

[0055] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, which is substituted with OR3, wherein R3is hydrogen or methyl.

[0056] In one embodiment of a compound for use according to the present invention:

[0057] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, which is substituted with SR3, wherein R3is hydrogen or methyl.

[0058] In one embodiment of a compound for use according to the present invention, the compound has formula la

[0059] R2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, or but-l-yne; or R2is Ci-Ce alkyl, preferably methyl.

[0060] In one embodiment of a compound for use according to the present invention, the compound has formula Ila

[0061] R2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, or but-l-yne.

[0062] In one embodiment of a compound for use according to the present invention, the compound has formula Ila

[0063] R2is Ci-Ce alkyl, preferably C1-C4 alkyl, more preferably methyl, or ethyl.

[0064] In one embodiment of a compound for use according to the present invention, the compound has formula Ila

[0065] R2is C2-C6 alkenyl, preferably C2-C5 alkenyl, more preferably allyl, vinyl, or dimethylallyl.

[0066] In one embodiment of a compound for use according to the present invention, the compound has formula Illa

[0067] R2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, or but-l-yne.

[0068] Preferably, the compound is selected from

[0069] MF -01: 1 -(2-Methoxyethyl)-2-methyl-5 -nitro- IH-imidazole :

[0070] MF -02 : 2-(Methoxymethyl)- 1 -methyl-5 -nitro- IH-imidazole :

[0071] MF-03: 1 -(2 -Ethoxy ethyl)-2-methyl-5-nitro-lH-imidazole: MF -04 : 2-(Ethoxymethyl)- 1 -methyl-5 -nitro- IH-imidazole :

[0072] MF-05: 2-Methyl-5-nitro-l -(2 -propoxy ethyl)- / / / -imidazole:

[0073] MF-06: l-Methyl-5-nitro-2-(propoxymethyl)-l / / -imidazole:

[0074] MF-07: l-(2-(Allyloxy)ethyl) -2-methyl-5-nitro-lH-imidazole:

[0075] MF-08 : 2-((Allyloxy)methyl)- 1 -methyl-5 -nitro- 177-imidazole :

[0076] MF-09: 2-Methyl-l-(2-((3-methylbut-2-en-l-yl)oxy)ethyl)-5-nitro-lH-imidazole:

[0077] Metro-P2 : 1 -(2-Methyl-5-nitro-7Z / -imidazol- 1 -yl)pent-4-yn-2-ol : Metro-P3 : 2-Methyl-5 -nitro- 1 -((prop-2-yn-l -yloxy)methyl)- IH-imidazole: and

[0078] Metro-Pl: 2-Methyl-5-nitro-l-(2-(prop-2-yn-l-yloxy)ethyl)-lH-imidazole:

[0079] More preferably, the compound is selected from and

[0080] (Metro-P2) (Metro-P3) (MF-01),

[0081] Preferably, the non-viral infections are infections with protozoa, anaerobic bacteria or microaerophilic bacteria.

[0082] Preferably, infections with protozoa are an infection with Trichomonas vaginalis, Giardia intestinalis (G. lambid), or Entamoeba histolytica.

[0083] More preferably, the non-viral infections are infections with anaerobic bacteria or microaerophilic bacteria.

[0084] Preferably, the non-viral infections are an infection with Helicobacter pylori, Clostridioides difficile, Fusobacterium nucleatum, or Gardnerella vaginalis.

[0085] In a preferred embodiment, the non-viral infection is an infection with a Helicobacter pylori strain having resistance against metronidazole. The compounds of the present invention preferably bind to thiol peroxidases whose inhibition may trigger toxic effects to bacteria. One of said thiol peroxidases is Helicobacter thiol peroxidase HpTpx (025151). The compounds of the present invention preferably also bind to H. pylori chaperonine HpGroEL (P42383).

[0086] Treatment methods

[0087] As discussed above, the present invention provides a method for the treatment of a non-viral infection or a disease caused by a non-viral infection.

[0088] Said method comprises the step of administering to a subject in need thereof a compound having general formula I or II or III according to the present invention or a pharmaceutical composition of the present invention.

[0089] Preferably, a therapeutically effective amount of a compound having general formula I or II or III according to the present invention the present invention or a pharmaceutical composition of the present invention is administered to the subject.

[0090] A “therapeutically amount” or “therapeutically effective amount”, both of which terms are used herein interchangeably, of a compound of the present invention (i.e. a compound having general formula I or II or III) or a pharmaceutical composition of the present invention is the amount which results in the desired therapeutic result.

[0091] The present invention also provides the use of the compound of the present invention (i.e. a compound having general formula I or II or III) or the use of the pharmaceutical composition of the present invention for the manufacture of a medicament.

[0092] Preferably, the non-viral infections are infections with protozoa, anaerobic bacteria or microaerophilic bacteria.

[0093] Preferably, infections with protozoa are an infection with Trichomonas vaginalis, Giardia intestinalis (G. lambid), or Entamoeba histolytica.

[0094] More preferably, the non-viral infections are infections with anaerobic bacteria or microaerophilic bacteria. Preferably, the infections with anaerobic bacteria or microaerophilic bacteria are an infection with Helicobacter pylori, Clostridioides difficile, Fusobacterium nucleatum, or Gardnerella vaginalis.

[0095] In a preferred embodiment, the non-viral infection is an infection with a Helicobacter pylori strain having resistance against metronidazole.

[0096] Nitroimidazole compounds

[0097] As outlined above, the present invention provides a compound having general formula III.

[0098] A compound having general formula III is a compound wherein

[0099] R1is Ci-Ce alkyl or Ci-Ce haloalkyl, and

[0100] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, wherein said Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl is optionally substituted with halogen, OR3or SR3, wherein R3is hydrogen or Ci-Ce alkyl, or a pharmaceutically acceptable salt thereof.

[0101] - Disclaimer

[0102] According to the invention, a compound of the present invention is not a compound wherein both R1and R2are methyl.

[0103] According to the invention, a compound of the present invention is not a compound with the following structure:

[0104] According to the invention, a compound of the present invention is not a compound wherein

[0105] R1is methyl and

[0106] R2is ethyl or C1-C4 alkyl substituted with halogen, OH, OMe, NH2, or NH(CH3)2.

[0107] According to the invention, a compound of the present invention is not a compound wherein R1is CF3 and

[0108] R2is C1-C4 alkyl substituted with halogen.

[0109] According to the invention, a compound of the present invention is not a compound wherein

[0110] R1is isopropyl and

[0111] R2is C1-C4 alkyl substituted with halogen.

[0112] According to the invention, a compound of the present invention is not a compound with any of the following structures:

[0113] In one embodiment, a compound of formula III has a structure wherein

[0114] R1is methyl and the compound has formula Illa In one embodiment, a compound of formula III has a structure wherein

[0115] R2is Ci-Ce alkyl, preferably C1-C4 alkyl, more preferably methyl, ethyl, propyl, isopropyl, n- propyl, n-butyl, isobutyl, sec-butyl, tert-butyl, neopentyl, or n-hexyl.

[0116] In one embodiment, a compound of formula III has a structure wherein

[0117] R2C2-C6 alkenyl, preferably C2-C5 alkenyl, more preferably allyl, vinyl, dimethylallyl, 1- butenyl, 2-butenyl, 3-butenyl, 1- pentenyl, 2-pentenyl, 3 -pentenyl, or 4-pentenyl.

[0118] In one embodiment, a compound of formula III has a structure wherein

[0119] R2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, but-l-yne, 1- butynyl, 2-butynyl, 3-butynyl, 1-propynyl, or 2-propynyl.

[0120] In one embodiment, a compound of formula III has a structure wherein

[0121] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, which is substituted with halogen.

[0122] In one embodiment, a compound of formula III has a structure wherein

[0123] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, which is substituted with OR3, and

[0124] R3is hydrogen or methyl.

[0125] In one embodiment, a compound of formula III has a structure wherein

[0126] R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, which is substituted with SR3, and R3is hydrogen or methyl.

[0127] In a preferred embodiment, the compound has formula Illa and

[0128] R2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, or but-l-yne.

[0129] A preferred compound having general formula III is

[0130] (Metro-P2: l -(2-Methyl-5-nitro- / / / -imidazol- l -yl)pent-4-yn-2-ol).

[0131] As discussed above, the present invention provides a pharmaceutical composition comprising

[0132] (i) at least one compound having general formula III according to the present invention,

[0133] (ii) optionally, pharmaceutical excipient(s) and / or carrier.

[0134] As discussed above, the present invention provides the compound having general formula III according to the present invention for use in medicine.

[0135] Pharmaceutical compositions or preparations may be selected according to the invention from the group of formulations containing tablets, layered tablets, coated tablets, pills, soft or hard capsules, microcapsules, oral retardant drug forms, transdermal systems, suppositories, micro- and nanocrystalline formulations, liposomal formulations, drops, nasal drops, sprays, emulsions, dispersions, solutions, sterile solutions, lyophilisates, powders and inhalation sprays.

[0136] The application or use of the pharmaceutical composition or preparation according to the invention is preferably selected from the group comprising oral, peroral, sublingual, buccal, subcutaneous, intravenous, dermal, pulmonary or nasal application or use.

[0137] Pharmaceutical compositions or preparations are preferably offered as sterile solutions or lyophilisates, parenteral, peroral and oral retardant drug forms, transdermal systems, micro- and nanocrystalline formulations, liposomal formulations, microcapsules, emulsions, dispersions and are particularly suitable for subcutaneous, intravenous, dermal, transdermal, oral, peroral or pulmonary use or application.

[0138] Lactose, starch, sorbitol, mannitol, sucrose, ethyl alcohol and water can be used, for example, as pharmacologically and chemically compatible carriers, solvents or auxiliary agents. Furthermore, starch, modified starch, gelatine, natural sugars, natural or synthetic polymers such as acacia gum, guar, sodium alginate, carboxymethyl cellulose or polyethylene glycol may be included as binders. Cyclodextrins, modified cyclodextrins, as well as benzoates, chlorides, acetates, tartrates may be contained as stabilizers and stearates, polyethylene glycol, amino acids such as leucine may be used as auxiliaries usually in concentrations of 0.05 % to 15 %.

[0139] Liquid formulations include solutions, dispersions and emulsions. Liquid formulations for parenteral use are sterile and contain water or water and solubilizers, such as propylene glycol, micelle and mixed micelle formers. Starch or modified starch, alginates, aluminates, bentonites or microcrystalline cellulose can be used as a liquid preparation usually in concentrations between 2% and 30% by weight.

[0140] Sugars, sugar alcohols, corn, rice or potato starch, gelatine, gum arabic, tragacanth sugar, ammonium calcium alginate carboxymethyl cellulose, hydroxy propyl methyl cellulose, polyvinyl pyrrolidone as well as inorganic substances can be used as auxiliary agents usually in concentrations between 1% and 30% by weight. Pharmaceutical preparations for subcutaneous, intravenous and transdermal use as well as parenteral and oral modified release dosage forms are claimed as preferred formulations. Such formulations usually consist of a matrix, in particular a matrix with polymers, in many cases biodegradable polymers as a shaping, constituent additive, into which at least one of the compounds of the present invention is incorporated.

[0141] The term "pharmaceutically acceptable salts" as used herein includes salts of the compound of the general formula III which are prepared with relatively nontoxic (i.e. pharmaceutically acceptable) acids or bases, depending on the particular substituents found on the compounds of the present invention. If, for example, compounds of the present invention contain acidic functionalities, base addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Non-limiting examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt. If compounds of the present invention contain basic functionalities, acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Non-limiting examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, phosphoric, partially neutralized phosphoric acids, sulfuric, partially neutralized sulfuric, hydroiodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p- tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like. Certain specific compounds of the present invention may contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts. Contacting the salt with a base may regenerate the neutral forms of the compounds of the present invention or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present invention. The compounds of the present invention may possess chiral or asymmetric carbon atoms (optical centers) and / or double bonds. The racemates, diastereomers, geometric isomers and individual optical isomers are encompassed by the present invention. The compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are also encompassed by the present invention. The compounds of the present invention may furthermore exist in multiple crystalline or amorphous forms.

[0142] The compounds of the present invention may further be in a so-called prodrug form. Prodrugs of the compounds of the invention are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Additionally, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex-vivo environment. For example, prodrugs can be slowly converted to the compounds of the present invention when, for example, placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0143] Further description of preferred embodiments

[0144] Novel antibiotic compounds are disclosed which are based on the nitroimidazole scaffold which exhibit >60-fold enhanced activity against Heliobacter pylori compared to the parent compound Metronidazole. In addition, they exhibit moderate activity in resistant isolates. The modifications to enhance activity occurred mainly at the free alcohol of the N-l hydroxy ethyl chain or at C2-ydroxymethyl chain to form an ether bond.

[0145] Mode of action studies point towards binding to the chaperonine HpgGroEL and enhanced binding to a thiol peroxidase (compared to Metronidazole) whose inhibition triggers toxic effects to bacteria due to inhibition of the overall oxidative stress response. The compounds are not toxic to human cells, exhibit suitable plasma stability, have favorable ADME (absorption, distribution, metabolism, excretion) and pharmacokintetic characteristics and are thus prime candidates for antibiotic use.

[0146] Herein nitroimidazole compounds with enhanced antimicrobial activity are disclosed to treat infections of H. pylori and other anaerobic or microaerophilic bacteria or protozoal infections. The cytotoxicity was evaluated in human cells which renders these compounds non-cytotoxic. The nitroimidazole compounds are thus potential drug candidates with a suitable safety profile.

[0147] The following examples and drawings illustrate the present invention without, however, limiting the same thereto.

[0148] BRIEF DESCRIPTION OF THE DRAWINGS

[0149] Figure 1 shows compounds of the present invention.

[0150] Figure 2 shows exemplarily results of MTT assay to determine IC50 values of compounds of the present invention in HeLa cells. Obtained Values after MTT readout of nitroimidazole compounds are normalized to DMSO control (DMSO Ctrl = 100% cell viability after 24 hours of incubation. MTT assays were performed within a concentration range from 10 pM to 1 mM.

[0151] Figure 3 shows in vitro stability of compounds of the present invention in mouse plasma.

[0152] Figure 4 shows modification of recombinantly expressed H. pylori thiol peroxidase (HpTpx) with metronidazole or Metro-P3 identified via intact-protein mass spectrometry after protein purification. H. pylori thiol peroxidase was overexpressed in E. coli in the presence of 500 pM metronidazole or Metro-P3, subsequently purified via affinity chromatography, analysed via IP -MS and compared to the unmodified HpTpx enzyme. Figure 5 shows the results of a peroxidaseassay with thiol peroxidase HpTpx. A decrease in activity can be seen with increasing the modification degree with Metro or Metro-P3.

[0153] Figure 6 shows IP-MS studies on improved efficacy of Metro-P3. Metro-P3 binds more readily (~8 fold) to HpTpx compared to Metronidazole showing the enhanced affinity of Metro- P3 to Metronidazole

[0154] Figure 7 shows different binding modes of metronidazole vs. Metro-P3 in crystallography studies with HpTpx. Metro-P3 binds to HpTpx in its reduced state which is the more stable and prominent form of HpTpx in the cytoplasm

[0155] Figure 8 shows pharmacokinetic data of different nitroimidazole compounds of the present invention.

[0156] EXAMPLES

[0157] Example 1 Materials and Methods

[0158] 1.1 Reagents and Solvents

[0159] Unless otherwise stated, commercially available reagents and starting materials are obtained from Sigma Aldrich, TCI Europe, VWR, Roth, BLDpharm and Alfa Aesar and starting compounds were used without further purification and stored as indicated. Technical solvents for purification were used after simple distillation.

[0160] All reactions sensitive to air and moisture were carried out using standard Schlenk techniques under argon atmosphere in flame-dried glassware. Anhydrous solvents and water-sensitive liquid chemicals were transferred using argon flushed syringes.

[0161] 1.2 Thin Layer Chromatography (TLC)

[0162] For thin layer chromatography (TLC) silica coated plates (aluminium, Merck, silica 60 F254) were used. For visualization, the spots were detected by using UV-light (254 nm and 366 nm) or by staining with a potassium permanganate solution (3.00 g KMNO4, 20.0 g K2CO3 and 5.00 mL 5% NaOH (aq.) in 300 mL water) followed by heat treatment. Column chromatography was carried out using silica gel (40-63 pm (Si 60)) from Merck.

[0163] 1.3 High Pressure Liquid Chromatography (HPLC) If indicated compounds were purified using preparative, reversed-phase HLPC using a Waters 2545 quaternary gradient module equipped with a fraction collector on an YMC Triart Cl 8 column (250 x 10 mm, 5 / / m). Gradient is listed in Table 1 using ddH2O and HPLC-grade acetonitrile (no TFA) as the mobile phase.

[0164] Table 1: Gradient used for HPLC purification. t (min) H2O (%) ACN (%)

[0165] 6 98 2

[0166] 1 98 2

[0167] 12 2 98

[0168] 13 2 98

[0169] 14 2 98

[0170] 15 98 2

[0171] 17 98 2

[0172] 1.4 Nuclear Magnetic Resonance Spectroscopy (NMR)

[0173] Nuclear Magnetic Resonance (NMR) spectra were measured at room temperature either on a Bruker AVHD-400 or AVHD-300. The chemical shifts are given in 3 values in ppm (parts per million) and calibrated to the residual proton signals of the solvents relative to the internal standard tetramethylsilane:

[0174] Chloroform-t / / (’H-NMR: 3 = 7.26 ppm,13C-NMR: 3 = 77.2 ppm)

[0175] The NMR multiplicities are given as singlets (s), doublets (d), triplets (t), quartets (q) , pseudosextets (ps) or multiplets (m). The coupling constants J are reported in Hertz (Hz). Assignments of ’H-NMR and13C-NMR signals are determined by two-dimensional NMR-spectroscopy (COSY, HSQC, HMBC).

[0176] 1.5 Mass Spectrometry (MS)

[0177] High-Resolution Mass Spectrometry (HR-MS) was performed on a LTQ-FT Ultra mass spectrometer (Thermo Fisher Scientific). ESI is used as an ionization method. Low resolution LC-MS measurements were conducted on a MSQ Plus mass spectrometer (Thermo Fisher Scientific). Processing of mass spectrometry data was done by Xcalibur 2.2 (Thermo Fisher Scientific). Example 2 Synthesis

[0178] 2.1 Metro-Pl (2-methyl-5-nitro- 1 -(2-(prop-2-yn- 1 -yloxy)ethyl)- IH-imidazole)

[0179] Metro-P1

[0180] To a suspension of metronidazole (200 mg, 1.17 mmol, 1.00 eq.) and CS2CO3 (838 mg, 2.57 mmol, 2.20 eq.) in DMF (4 mL) is added propargyl bromide solution (80% in toluene, 0.30 mL, 2.78 mmol, 2.40 eq.) and heated to 50 °C for 96 h. The reaction mixture is quenched via addition of H2O (5 mL) and extracted with EtOAc (3 x 20 mL). Combined organic layers are subsequently washed with 5% LiCl solution (aq., 15 mL) and brine (20 mL), dried over Na2SO4 and the solvent is removed in vacuo. The crude product is further purified performing column chromatography (EtOAc 100%) and high-performance liquid chromatography (HPLC) to obtain 74.4 mg of the desired probe (30%, 1.17 mmol) as a white solid.

[0181] TLC: Rf= 0.45 (EtOAc) [UV],

[0182] 'H NMR (400 MHz, CDCh) 8 (ppm) = 2.39 (t, = 2.3 Hz, 1H, H-12), 2.52 (s, 3H, H-6), 3.86 (t,3J = 4.9 Hz, 2H, H-8), 4.08 (d, = 2.3 Hz, 2H, H-10), 4.51 (t,3J = 4.9 Hz, 2H, H-7), 7.96 (s, 1H, H-4).

[0183] 13C NMR (101 MHz, CDCh) 6 (ppm) = 14.8 (C-6), 46.6 (C-7), 58.7 (C-10), 68.6 (C-8), 75.3 (C-12), 78.8 (C-l l), 133.4 (2 x C, C-4 / C-5) 152.0 (C-2).

[0184] HR-MS (ESI): m / z = calc. [M+H]+: 210.0878, found: 210.0874.

[0185] 2.2 2-(2-Methyl-5-nitro- / / / -imidazol- 1-yl (acetaldehyde (part of Metro-P2 synthesis)

[0186] To 160 mL of CH2Q2 is added dropwise 2.00 mL (20.0 mmol, 1.10 eq.) of oxalyl chloride under Argon atmosphere. The solution is cooled to -78 °C and 10 mL of DMSO is added dropwise to the stirred solution. 20 min later, 3.42 g (20 mmol, 1.00 eq.) of metronidazole dissolved in 15 mL of DMSO is added. After 20 min of additional stirring, 33 mL (240 mmol, 12.0 eq.) of NEt3is added. The reaction mixture is stirred for another 10 min at -78 °C and is then allowed to warm to room temperature. The mixture is diluted with EtOAc (400 mL) and washed with water (4 * 75 mL). The water phase is extracted with EtOAc (3 x 50 mL). The combined organic layers are washed with brine (150 mL), dried over Na2SO4 and the solvent is removed in vacuo. The resulting crude residue is purified by flash silica gel chromatography (CH2Cl2 / MeOH 40: 1 v / v) to give the target compound (1.20 g, 7.09 mmol, 32%) as an orange, viscous oil.

[0187] TLC: Rf= 0.78 (CH2Cl2 / MeOH 10: 1) [UV],

[0188] 'H NMR (400 MHz, CDCh) 8 (ppm) = 2.41 (s, 3H, H-6), 5.21 (s, 2H, H-7), 7.99 (s, 1H, H- 4), 9.75 (s, 1H, H-8).

[0189] 13C NMR (101 MHz, CDCh) 6 (ppm) = 14.0 (C-6), 54.8 (C-7), 132.8 (C-4), 132.9 (C-5), 150.4 (C-2), 191.6 (C-9).

[0190] LR-MS: m / z = calc. [M+H]+: 170.06, found: 170.11.

[0191] Metro-P2 ( I -(2-Methyl-5-nitro- / 7 / -imidazol - I -yl)pent-4-yn-2-ol)

[0192] Metro-P2

[0193] A mixture of 620 mg (25.5 mmol, 1.78 eq.) of magnesium turnings, 3.56 g (15.8 mmol, 1.10 eq.) of ZnBr2and iodine (5 mol%) in dry THF (5 mL) is stirred for 15 min at room temperature. Then, a solution of propargylbromide (1.55 mL, 80% in toluene, 14.4 mmol, 1.00 eq.) in dry THF (15 mL) is added dropwise. When the reaction mixture starts to reflux, it is cooled to 0 °C. After complete addition of the bromide, the reaction mixture is stirred for 1 h at room temperature. A full conversion is assumed and the crude material is directly used in the subsequent step.

[0194] The metronidazole-aldehyde (500 mg, 2.96 mmol, 1.00 eq.) is dissolved in dry THF (8 mL) and 5.30 mL (3.84 mmol, 1.30 eq.) of previously synthesized Grignard reagent is added. The mixture is stirred for 2 h at room temperature and then poured into ice water. Saturated aqueous NH4CI solution (20 mL) is added to dissolve the precipitate and the organic layer is separated. The aqueous layer is extracted with Et2O (3 x 30 mL). The combined organic extracts are washed with brine (2 x 20 mL) and dried over Na2SO4. After evaporation of the solvent the residue is purified by column chromatography (EtOAc 100%) and HPLC to obtain 34 mg (5%, 0.16 mmol) of the desired probe Metro-P2.

[0195] TLC: Rf= 0.41 (EtOAc) [UV],

[0196] 'H NMR (300 MHz, CDCh) 8 (ppm) = 2.18 (t, = 2.7 Hz, 1H, H-12), 2.56 - 2.60 (m, 2H, H-10), 2.72 (s, 3H, H-6), 4.14 - 4.36 (m, 2H, H-8, H-9) 4.77 (d,3J= 11.8 Hz, 1H, H-8), 8.06 (s, 1H, H-4).

[0197] 13C NMR (75 MHz, CDCh) 6 (ppm) = 14.6 (C-6), 25.4 (C-10), 50.7 (C-8), 69.0 (C-9), 72.3 (C-12), 78.5 (C-l l), 132.7 (C-4), 140.2 (C-5), 152.1 (C-2).

[0198] HR-MS (ESI): m / z = calc. [M+H]+: 210.0878, found: 210.0874.

[0199] 2.3 Metro-P3 (2-methyl-5-nitro- 1 -((prop-2 -yn- 1 -yloxy)methyl)- IH-imidazole)

[0200] Metro-P3

[0201] To a suspension of (2-methyl-5-nitro-7J7-imidazol-l-yl)methanol (250 mg, 1.59 mmol, 1.00 eq.) and CS2CO3 (1.20 g, 3.91 mmol, 2.30 eq.) in THF (10 mL) is added propargyl bromide solution (80% in toluene, 0.40 mL, 3.66 mmol, 1.20 eq.) and heated to 70 °C for 96 h. The reaction mixture is quenched via addition of 6H2O (5 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers are subsequently washed with brine (20 mL) and water (20 mL), dried over Na2SO4 and the solvent is removed in vacuo. The crude product is further purified performing column chromatography (hexane / EtOAc 1 : 1) and high-performance liquid chromatography (HPLC) to obtain 136.5 mg of the desired probe Metro-P3 (44%, 0.70 mmol) as a white solid.

[0202] TLC: Rf= 0.36 (EtOAc) [UV],

[0203] 'H NMR (300 MHz, CDCh) 6 (ppm) = 2.51 (t, = 2.4 Hz, 1H, H-l 1), 4.05 (s, 3H, H-6), 4.25 (d, = 2.4 Hz, 2H, H-9), 4.79 (s, 2H, H-7), 7.97 (s, 1H, H-4).

[0204] 13C NMR (75 MHz, CDCh) 6 (ppm) = 34.2 (C-6), 58.5 (C-9), 63.2 (C-7), 76.2 (C-l 1), 78.2 (C-10), 129.6 (2 x C, C-4, C-5), 147.8 (C-2). HR-MS (ESI): m / z = calc. [M+H]+: 196.0717, found: 196.0715.

[0205] 2.4 MF-01 (l-(2-Methoxyethyl)-2-methyl-5-nitro-lH-imidazole)

[0206] MF-01

[0207] CS2CO3 (1.71 g, 5.28 mmol, 4.50 eq.) is added to metronidazole (200 mg, 1.17 mmol, 1.00 eq.) dissolved in THF (10 mL) and stirred at room temperature for 30 min. Methyl iodide (582 pL, 1.33 g, 9.36 mmol, 8.00 eq.) is added and the solution stirred at room temperature for 21 h. The solvent is removed under reduced pressure, the remaining residue is dissolved in EtOAc and then filtered. The crude product is purified by flash silica chromatography (SiCh, hexane / EtOAc = 1 / 1, 2.5 x 20 cm) and HPLC to afford MF-01 (28.4 mg, 153 pmol, 13 %) as a white solid.

[0208] TLC: Rf= 0.35 (EtOAc) [UV],

[0209] HPLC: / R = 5.0 min

[0210] *H-NMR (400 MHz, CDCh): 6 [ppm] = 2.75 (s, 3H, H-6), 3.30 (s, 3H, H-9), 3.73 (t,3J= 4.8 Hz, 2H, H-8), 4.59 (t,3J= 4.8 Hz, 2H, H-7), 8.07 (s, 1H, H-4).

[0211] 13C-NMR (101 MHz, CDCh): 6 [ppm] = 13.6 (q, C-6), 47.6 (t, C-7), 59.4 (q, C-9), 70.7 (t, C-8), 126.9 (d, C-4), 136.6 (s, C-5), 150.8 (s, C-2).

[0212] HR-MS (ESI): m / z = calc. [M+H]+: 186.0873, found: 186.0871.

[0213] 2.5 MF-02 (2-(Methoxymethyl)-l-methyl-5-nitro-lH-imidazole)

[0214] MF-02 CS2CO3 (1.04 g, 3.18 mmol, 2.50 eq.) is added to (l-methyl-5-nitro-7J / -imidazol-2-yl)methanol (200 mg, 1.27 mmol, 1.00 eq.) dissolved in THF (10 mL) and stirred at room temperature for 30 min. Methyl iodide (560 pL, 1.26 g, 8.89 mmol, 7.00 eq.) is added and the solution is stirred at room temperature for 21 h. The solvent is removed under reduced pressure, the remaining residue is dissolved in EtOAc and then filtered. The crude product is purified by flash silica chromatography (SiCh, hexane / EtOAc = 1 / 1) and HPLC to afford MF-02 (84.8 mg, 495 pmol, 39 %) as a white solid.

[0215] TLC: Rf= 0.48 (EtOAc) [UV],

[0216] HPLC: / R = 6.6 min.

[0217] *H-NMR (400 MHz, CDCh): 6 [ppm] = 3.41 (s, 3H, H-8), 4.04 (s, 3H, H-6), 4.65 (s, 2H, H- 7), 7.98 (s, 1H, H-4).

[0218] 13C-NMR (101 MHz, CDCh): 6 [ppm] = 34.4 (q, C-6), 58.9 (q, C-8), 66.3 (t, C-7), 130.2 (d, C-4), 148.5 (s, C-5), 157.3 (s, C-2).

[0219] 2.6 MF-03 (l-(2-ethoxyethyl)-2-methyl-5 -nitro- IH-imidazole)

[0220] MF-03

[0221] 200 mg of metronidazole (1.00 eq., 1.17 mmol) and 457 mg of caesium carbonate (1.20 eq.,

[0222] 1.40 mmol) are suspended in 5 mL dry THF. To this suspension, 0.20 ml of ethyl bromide (2.40 eq., 306 mg, 2.80 mmol) are slowly added and then heated to reflux. Over a period of 60 h, 4.80 equivalents of ethyl bromide (612 mg, 5.60 mmol) and one equivalent of caesium carbonate (380 mg, 1.16 mmol) are added. The suspension is quenched with 20 mL dest. water. The reaction mixture is extracted with ethyl acetate (3 x 20 mL). The combined organic layers are washed with 20 mL of saturated NaCl solution and dried over sodium sulfate. The excess solvent is removed in vacuo. The crude product is separated chromatographically (SiCh, hex / EtOAc = 1 :5) to obtain an orange-brown solid (37.8 mg, 0.19 mmol, 16%).

[0223] TLC: Rf= 0.30 (hexane / EtOAc 1 :5) [UV],

[0224] *H-NMR (400 MHz, CDCh): 8 [ppm] = 1.10 (t,3J= 7.0 Hz, 3H, H-10), 2.58 (s, 3H, H-6),

[0225] 3.41 (q,3J = 7.0 Hz, 2H, H-9), 3.73 (t,3J= 5.0 Hz, 2H, H-8), 4.51 (t,3J= 5.0 Hz, 2H, H-7), 7.98 (s, 1H, H-4).13C-NMR (100 MHZ, CDCh): 6 [ppm] = 14.4 (C-6), 15.1 (C-10), 47.1 (C-7), 67.1 (C-9), 69.1 (C-8), 131.6 (C-4), 138.4 (C-5), 151.7 (C-2).

[0226] HR-MS (ESI): m / z = calc. [M+H]+: 200.1030, found: 200.1029.

[0227] 2.7 MF-04 2-(Ethoxymethyl)-l-methyl-5-nitro-lH-imidazole (MF-RP-09)

[0228] MF-04

[0229] CS2CO3 (2.49 g, 7.64 mmol, 4.00 eq.) and tetrabutylammonium iodide (TBAI, 317 mg, 0.45 mol%) are added to (l-methyl-5-nitro-7Z / -imidazol-2-yl)methanol (300 mg, 1.91 mmol, 1.00 eq.) dissolved in THF (10 mL) and stirred at room temperature for 30 min. Ethyl bromide (0.56 mL, 7.64 mmol, 4.00 eq.) is added and the solution is stirred at reflux for 21 h. The reaction mixture is filtered, and the solvent removed under reduced pressure. The crude product is dissolved in EtOAc, filtered and purified by flash silica chromatography (SiCh, hexane / EtOAc = 1 / 2) followed by HPLC purification. MF-04 is obtained pure as a yellow oil (80.0 mg, 1.11 mmol, 58%).

[0230] TLC: Rf= 0.44 (hexane / EtOAc 2: 1) [UV],

[0231] HPLC: fe = 7.7 min

[0232] 'H NMR (400 MHz, CDCh) 8 [ppm] = 1.22 (t,3J = 7.1 Hz, 3H, H-9), 3.57 (q,3J= 7.1 Hz, 2H, H-8), 4.04 (s, 3H), 4.66 (s, 2H), 7.95 (s, 1H).

[0233] 13C NMR (100 MHz, CDCh) 6 = 15.1 (C-9), 33.9 (C-6), 64.7 (C-8), 66.8 (C-7), 130.8 (C-4), 139.7 (C-5), 149.0 (C-2).

[0234] HR-MS (ESI): m / z = calc. [M+H]+: 186.0873, found: 186.0872.

[0235] 2.8 MF-05 (2-methyl-5 -nitro- 1 -(2 -propoxy ethyl)- / / / -imidazole)

[0236] MF-05

[0237] CS2CO3 (1.14 g, 3.51 mmol, 4.99 eq.) is added to metronidazole (150 mg, 876 pmol, 1.00 eq.) dissolved in THF (10 mL) and stirred at room temperature for 30 min. Propyl iodide (0.85 mL, 8.76 mmol, 10.0 eq.) is added and the solution is stirred at reflux overnight. The reaction mixture is filtered and the solvent removed under reduced pressure. The crude product is purified by flash silica chromatography (SiCh, hexane / EtOAc = 1 / 2) to afford MF-05 (38.5 mg, 184 pmol, 21%) as a yellow oil.

[0238] TLC: Rf= 0.23 (hexane / EtOAc 1:2) [UV],

[0239] 'H NMR (400 MHz, CDCh) 6 [ppm] = 0.83 (t,3J = 7.1 Hz, 3H, H-l l), 1.49 (ps,3J = 7.1 Hz, 2H, H-10), 2.58 (s, 3H, H-6), 3.31 (t,3J= 7.1 Hz, 2H, H-9), 3.73 (t,3J= 4.5 Hz, 2H, H-8), 4.52 (t,3J= 4.5 Hz, 2H, H-7), 7.99 (s, 1H, H-4).

[0240] 13C NMR (100 MHz, CDCh) 6 [ppm] = 10.6 (C-l l), 14.4 (C-6), 22.9 (C-10), 47.1 (C-7), 69.3 (C-9), 73.4 (C-8), 131.9 (C-4), 138.3 (C-5), 151.6 (C-2).

[0241] HR-MS (ESI): m / z = calc. [M+H]+: 214.1186, found: 214.1184.

[0242] 2.9 MF-06 (l-methyl-5-nitro-2-(propoxymethyl)-U / -imidazole)

[0243] CS2CO3 (3.32 g, 7.64 mmol, 4.00 eq.) is added to (l-methyl-5-nitro-7J / -imidazol-2-yl)methanol 400 mg, 2.55 mmol, 1.00 eq.) dissolved in THF (10 mL) and stirred at room temperature for 30 min. Propyl iodide (1.59 mL, 15.3 mmol, 6.00 eq.) is added and the solution is stirred at reflux over night. The reaction mixture is filtered, and the solvent removed under reduced pressure. The crude product is purified by flash silica chromatography (SiCh, hexane / EtOAc = 1 / 2) to obtain MF-06 pure as a yellow oil (57.6 mg, 281 pmol, 11%).

[0244] TLC: Rf= 0.57 (hexane / EtOAc 1:3) [UV],

[0245] 'H NMR (400 MHz, CDCh) 6 [ppm] = 0.91 (t,3J = 7.2 Hz, 3H, H-10), 1.61 (ps,3J = 7.2 Hz, 2H, H-9), 3.44 (t,3J= 7.2 Hz, 2H, H-8), 4.02 (s, 3H, H-6), 4.62 (s, 2H, H-7), 7.93 (s, 1H, H-4).

[0246] 13C NMR (100 MHz, CDCh) 6 [ppm] = 10.6 (C-10), 22.9 (C-9), 33.8 (C-6), 65.3 (C-8), 73.0 (C-7), 131.6 (C-4, C-5), 149.2 (C-2).

[0247] HR-MS (ESI): m / z = calc. [M+H]+: 200.1030, found: 200.1028.

[0248] 2.10 MF-07 (l-(2-(allyloxy)ethyl) -2-methyl-5-nitro-lH-imidazole)

[0249] MF-07

[0250] 200 mg of metronidazole (1.17 mmol, 1.00 eq.) and 762 mg of caesium carbonate (2.34 mmol, 2.00 eq.) are suspended in 10 mL dry THF. To this suspension, 0.30 ml of allyl bromide (.2.80 mmol, 3.00 eq.) is slowly added and then heated to 70°C. After 23 h, the suspension is quenched with 20 mL of dest. water. The reaction mixture is extracted with ethyl acetate (3 x 20 mL). The combined organic layers are washed with 20 mL of saturated NaCl solution and dried over sodium sulfate. The excess solvent is removed in vacuo. The crude product is separated chromatographically (1 :5 hexane to ethyl acetate) to obtain MF-07 pure as a gold-brown solid (74.5 mg, 0.35 mmol, 30%).

[0251] TLC: Rf= 0.40 (hexane / EtOAc 1 :5) [UV],

[0252] 'H NMR (400 MHz, CDCh) 8 [ppm] = 2.57 (d,3J= 2.6 Hz, 3H, H-6), 3.75 (t,3J= 5.1 Hz, 2H, H-8), 3.84 - 3.93 (m, 2H, H-9), 4.52 (t,3J= 5.1 Hz, 2H, H-7), 5.09 - 5.20 (m, 2H, H-l l), 5.66 - 5.81 (m, 1H, H-10), 7.98 (d,3J= 2.6 Hz, 1H, H-4).

[0253] 13C-NMR (100 MHZ, CDCh): 6 [ppm] = 14.5 (C-6), 46.9 (C-7), 68.7 (C-8), 72.3 (C-9), 117.7 (C-l l), 132.0 (C-4), 133.8 (C-10), 138.4 (C-5), 151.7 (C-2).

[0254] HR-MS (ESI): m / z = calc. [M+H]+: 212.1030, found: 212.1030. 2.11 MF-08 (2-((allyloxy)methyl)-l-methyl-5 -nitro- 177-imidazole)

[0255] CS2CO3 (1.56 g, 4.77 mmol, 3.00 eq.) is added to (l-methyl-5-nitro-7J / -imidazol-2-yl)methanol 250 mg, 1.59 mmol, 1.00 eq.) dissolved in THF (12 mL) and stirred at room temperature for 30 min. Allyl iodide (0.44 mL, 4.77 mmol, 3.00 eq.) is added and the solution is stirred at reflux for 3 days. The reaction mixture is filtered, and the solvent removed under reduced pressure. The crude product is purified by flash silica chromatography (SiCh, hexane / EtOAc = 1 / 1) and HPLC to obtain MF-08 pure as a colourless oil (138 mg, 700 pmol, 44%).

[0256] TLC: Rf= 0.40 (hexane / EtOAc 1: 1) [UV],

[0257] 'H NMR (400 MHz, CDCh) 8 [ppm] = 4.02 - 4.06 (m, 5H, H-6, H-8), 4.66 (s, 2H, H-7), 5.20 - 5.37 (m, 2H, H-10), 5.77 - 5.96 (m, 1H, H-9)), 7.95 (s, 1H, H-4).

[0258] 13C NMR (100 MHz, CDCh) 6 [ppm] = 33.9 (C-6), 64.1 (C-8), 72.0 (C-7), 118.8 (C-10), 131.0 (C4, C5), 133.3 (C-9), 148.8 (C-2).

[0259] HR-MS (ESI): m / z = calc. [M+H]+: 198.0873, found: 198.0872.

[0260] 2.12 MF-09 (2-methyl-l-(2-((3-methylbut-2-en-l-yl)oxy)ethyl)-5-nitro-lH-imidazole)

[0261] MF-09

[0262] 200 mg metronidazole (1.17 mmol, 1.00 eq.) and 762 mg of caesium carbonate (2.34 mmol, 2.00 eq.) are suspended in 9 mL dry THF. To this reaction mixture, 0.40 ml 3,3-dimethylallyl bromide (3.51 mmol, 3.00 eq.) are slowly added and then heated to reflux. After 18 h, the suspension is quenched with 20 mL of dest. water. The reaction mixture is extracted with ethyl acetate (3 x 20 mL). The combined organic layers are washed with 20 mL of saturated NaCl solution and dried over sodium sulfate. The excess solvent is removed in vacuo. The crude product is separated chromatographically (hexane / EtOAc) followed by HPLC purification. The desired product MF-08 is obtained pure as a white solid remained (29.0 mg, 0.12 mmol, 10%). TLC: Rf= 0.42 (hexane / EtOAc 1 :5) [UV],

[0263] *H-NMR (400 MHz, CDCh): 8 [ppm] = 1.17 (s, 3H, H-13), 1.76 (s, 3H, H-12), 2.52 (s, 3H, H-6), 4.47 (t,3J= 5.0 Hz, 2H, H-8), 4.59 (m,3J= 7.3 Hz, 1.5 Hz, 4H, H-9, H-7), 5.32 (tt, 1H, H-10), 7.98 (s, 1H, H-4).

[0264] 13C-NMR (100 MHZ, CDCh): 6 [ppm] = 14.2 (C-6), 18.1 (C-12), 25.8 (C-13), 45.4 (C-7), 65.3 (C-9), 65.6 (C-8), 117.5 (C-10), 130.1 (C-4), 132.6 (C-l l), 140.9 (C-5), 151.1 (C-2). HR-MS (ESI): m / z = calc. [M+H]+: 240.1343, found: 240.1343.

[0265] Example 3 Biological testing

[0266] 3.1 Inhibition assays

[0267] The compounds of the present invention were tested against numerous bacterial strains. Tables 2 and 3 below summarize the results and indicate the minimal inhibitory concentration (MIC) values of the compounds against the indicated bacterial strains.

[0268] Table 2: Measured MIC values of metronidazole, Metro-Pl, Metro-P2 and Metro-P3 in different bacterial strains: H. pylori (microaerophilic), C. difficile (anaerobic), E. coli (aerobic), S. aureus (aerobic). Read out of MIC assays after 3 days of incubation (H. pylori and C. difficile') and after one day (E coli and S. aureus). Assay was performed in duplicates.

[0269] MIC values H. pylori C. difficile E. coli S. aureus

[0270] Metronidazole 12.5 - 50pM 1 pM >1 mM >1 mM

[0271] Metro-Pl 190 nM 6.25 pM >1 mM >1 mM

[0272] Metro-P2 12.5 - 50 pM 1 pM >1 mM >1 mM

[0273] Metro-P3 250 - 380 nM 6.25 pM >1 mM >1 mM

[0274] In H. pylori, the determined MIC of metronidazole (12.5 - 50 pM) was within the literature MIC range of 10 to 50 pM for susceptible strains (Megraud 2007; Wu et al., 2000), which is important for the accuracy and comparability of the obtained MIC values in this assay among each other. The MIC value of Metro-P2 (12.5 - 50 pM pM) was identical to the one of metronidazole. Surprisingly, the MIC values of Metro-Pl (250 nM - 380 nM nM) and Metro- P3 (190 - 250 nM nM) were up tolOO fold more potent than the determined value for metronidazole (12.5 - 50 pM). A huge increase in antibiotic activity was observable, which may be attributed to the modification of the free hydroxyl group to the ether moiety in both probes. Therefore, several derivatives were synthesized and additionally assessed for their antimicrobial activity in susceptible and resistant / / , pylori strains.

[0275] MIC assays were additionally performed for susceptible and metronidazole resistant H. pylori strains within a concentration range from 100 nM to 1 mM. The MIC assay for susceptible H. pylori was performed twice and visual read out of the assay was done after 3 days of incubation with metronidazole or respective probes. The resulting values are summarized in Table 3.

[0276] Table 3: Measured MIC values of metronidazole, Metro-Pl, Metro-P2, Metro-P3, MF01

[0277] - MF09 in H. pylori (wildtype, ATCC 26695) and a metronidazole-resistant H. pylori strain (clinical isolate). Read out of MIC assays after 3 days of incubation. MIC assays were performed within a concentration range from 90 nM to 50 pM for susceptible strains and from

[0278] 950 nM to 500 pM.

[0279] MIC (H. pylori MIC (H. pylori

[0280] Compound

[0281] 26695) metro-res)

[0282] Dimetridazole- 125-250 pM

[0283] OH

[0284] Metronidazole 12.5 pM 250 pM

[0285] Metro-Pl 390 nM 125 pM

[0286] Metro-P2 3.13 pM 250 - 500 pM

[0287] Metro-P3 195 nM 62.5 pM

[0288] MF01 780 nM 125 pM

[0289] MF02 390 nM 125 pM

[0290] MF03 390 nM 62.5 - 125 pM

[0291] MF04 390 nM 125 pM

[0292] MF05 390 nM 62.5 pM

[0293] MF06 390 nM 62.5 pM

[0294] MF07 390 nM 125 pM

[0295] MF08 390 nM 62.5 pM

[0296] MF09 1.56 -3.13 pM 125 pM

[0297] A MIC shift for metronidazole and all tested compounds against drug resistant H. pylori was observed, which is an indication for similar mechanisms of resistance.. However, Metro-P3, MF03, MF05, MF06, MF08 still had better MIC values (62.5 pM) in resistant H. pylori than metronidazole and Metro-P2 (250 - 500 pM) and were close to the lower limit of MIC range of metronidazole in susceptible / / , pylori strains (10 - 50 pM) (Megraud 2007; Wu etal., 2000). 3.2 Target identification

[0298] Target identification experiments showed that to-date two unknown protein targets of metronidazole in H. pylori are also involved in the mechanism of action of these compounds, namely:

[0299] - H. pylori chaperonine HpGroEL (P42383), and

[0300] - the thiol peroxidase HpTpx (025151).

[0301] MS based binding site studies identified a hydroxylamine intermediate of metronidazole-probes bound to the active site cysteine of these proteins as a suicide inhibitor. Additionally, HpTpx and HpGroEL could be successfully modified with metronidazole or Metro-P3 during recombinant protein expression (exemplarily shown for HpTpx in Figure 4). Further functional activity assays for both enzymes showed reduced or complete loss of activity: ATPase activity assays showed up to 60% reduction of HpGroEL upon modification with metronidazole or Metro-P3. Antioxidant activity assays showed modification-dependent loss of activity if metronidazole or Metro-P3 are covalently bound to HpTpx in the active site rendering H. pylori in total less effective to eliminate oxidative stress leading to cell death. However, most interesting is that the novel compounds e.g Metro-P3 modify HpTpx more readily than Metronidazole (as shown in vitro during recombinant protein expression in E. coH. Figure 6 and in-situ in H. pylori).

[0302] 3.3 Crystallography

[0303] Crystallography also revealed completely different binding motifs of Metronidazole and Metro-P3 to HpTpx which additionally showcases that even minor changes to the structure lead to significant alterations in the binding affinity and mode of action of nitroimidazoles in H. pylori (Figure 7). Metro-P3 binds to HpTpx in its reduced state (which is the more stable and more prevalent form in the cytoplasm in H. pylori, Figure 7B), and forms a helix dipole to stabilize the ligand, whereas Metronidazole binds to HpTpx shortly after the disulfide bridge is openend in the catalytic cycle (Figure 7A). Forming the active site disulfide bridge in HpTpx leads to constrained geometry in the protein (cysteines are far apart from each other) making the oxidized state of HpTpx less thermodynamically stable. Due to the different binding motifs we also have additional proof that Metro-P3 is more favoured to inhibit the activity of HpTpx and support our invention that small changes in our structures do have a significant change in mode of action compared to the current gold standard metronidazole. In summary, HpTpx is mainly contributing to the activity increase of our novel nitroimidazole compounds (which exhibits >60-fold enhanced activity against Heliobacter pylori compared to the parent compound Metronidazole.

[0304] 3.4 Cell toxicity

[0305] Mohindra and Rauth (1976) published an IC50 value >10 mM for metronidazole in HeLa cells after 14 days of incubation without media change rendering metronidazole a non-toxic drug for human cells. However, the incubation time was set very high in this experiment and as Metro- Pl and Metro-P3 are up to 100 fold more potent in H. pylori than metronidazole it was important to know if this boost in antimicrobial activity is also directly linked to an increase in human cell toxicity. Therefore, MTT assays in HeLa and HepG2 cells were performed for different concentrations of compound (range 1 pM to 1 mM) with an incubation time set to24 hours.

[0306] Table 4: Measured IC50 values after MTT readout of nitroimidazole compounds. Read out of MTT assays after 24 hours of incubation. MTT assays were performed within a concentration range from 10 pM to 1 mM for both cell lines (HeLa and HepG2).

[0307] MTT MTT

[0308] Compound (HeLa) (HepG2)

[0309] Metronidazole > 1 mM n.d.

[0310] Metro-Pl > 1 mM > 1 mM

[0311] Metro-P2 > 1 mM > 1 mM

[0312] Metro-P3 > 1 mM > 1 mM

[0313] MF01 > 1 mM > 1 mM

[0314] MF02 > 1 mM > 1 mM

[0315] MF03 > 1 mM > 1 mM

[0316] MF04 > 1 mM > 1 mM

[0317] MF05 > 1 mM > 1 mM

[0318] MF06 > 1 mM > 1 mM

[0319] MF07 > 1 mM > 1 mM

[0320] MF08 > 1 mM > 1 mM

[0321] MF09 > 1 mM > 1 mM

[0322] After read-out of the assay, no IC50 values could be determined up to a final concentration of 1 mM for all tested compounds in HeLa and HepG2 cell lines since no decrease in metabolic activity was observable. Only for Metro-Pl a slightly diminished metabolic activity could be observed for a final concentration of 10 mM and Metro-Pl started to impose cytotoxicity for HeLa cells at concentrations >10 mM. But it was not possible to determine the exact IC50 value due to the insolubility of Metro-Pl in DMSO in higher concentrations. IC50 values for compounds are summarized in table 4. A few selected examples are shown in Figure 2.

[0323] 3.5 Plasma stability

[0324] We additionally performed in vitro plasma stability assays in mouse serum for compounds Metro-Pl, Metro-P2, Metro-P3, MF01 to MF09 for 28 hours. All tested compounds showed very good stability and no half-life could be determined. Exemplarily, after 28h, 75%% of Metro-Pl, 99% of Metro-P2, 82% of Metro-P3, 66% of MF01, 81% of MF03 and 87% of MF07 were still present in the sample and not decomposed (see Figure 3).

[0325] Table 5 Overview of all in vitro data. Metronidazole is the starting material for synthesized compounds Metro-Pl, Metro-P2, MF01, MF03, MF05, MF07 and MF09. Dimetridazole-OH is the starting material for synthesized compounds Metro-P3, MF02, MF04, MF06 and MF08. Both are added for reference.

[0326] MICMICMTTMTTPlasma

[0327] Dimetridazole- _ , _ . . , a , ,

[0328] 3.13 pM 780 nM > 1 mM . n.d. n.d.

[0329] Oil

[0330] Metronidazole 12.5 pM 3.125 pM > 1 mM n.d.

[0331] Metro-Pl 390 nM < 97 nM > 1 mM > 1 mM

[0332] Metro-P2 3.13 pM 780 nM - 1.56 pM > 1 mM > 1 mM

[0333] Metro-P3 195 nM 195 nM - 390 nM > 1 mM > 1 mM

[0334] MF-01 780 nM < 97 nM > 1 mM > 1 mM

[0335] MF-02 390 nM 195 nM - 390 nM > 1 mM > 1 mM

[0336] MF-03 390 nM < 97 nM > 1 mM > 1 mM

[0337] MF-04 390 nM 390 nM > 1 mM > 1 mM

[0338] MF-05 390 nM < 97 nM > 1 mM > 1 mM

[0339] MF-06 390 nM 390 nM > 1 mM > 1 mM

[0340] MF-07 390 nM < 97 nM > 1 mM > 1 mM

[0341] MF-08 390 nM 390 nM > 1 mM > 1 mM

[0342] MF-09 1.56 -3.13 pM 390 nM > 1 mM > 1 mM

[0343] 3.6 ADME (absorption, distribution, metabolism, and excretion studies)

[0344] Characterization of absorption, distribution, metabolism, and excretion (ADME) properties was performed for 6 potential candidates (Metro-Pl, Metro-P3, MF-01, MF-02, MF-03 and MF-07) to provide further safety considerations of the nitroimidazole compounds of the present invention. 4 out of 6 compounds achieve necessary ADME threshold values showcasing that the compounds of the present invention do hold immense potential for further drug development (Table 6).

[0345] Table 6 Overview of ADME and protein plasma binding data: 6 candidates (Metro-Pl, Metro-P3, MF01, MF02, MF03, MF07) were in vitro tested for ADME properties and protein plasma binding to human or mouse plasma. mouse Mouse human Human Mous Huma PPB PPB

[0346] Half-life Clearance Half-life Clearance e n Mous Huma microsome [pl / min / m microsome [pl / min / m plasm plasm e [%] n [%] s [min] g protein] s [min] g protein] a a

[0347] Half- Halflife life

[0348] [min] [min]

[0349] Metro 17.3 80.0 11.2 124.0 > 240 > 240 66.2 68.9

[0350] -Pl

[0351] Metro > 60 < 23 > 60 < 23 > 240 > 240 45.1 50.0

[0352] -P3

[0353] MF-01 > 60 < 23 > 60 < 23 > 240 > 240 76.7 74.4

[0354] MF-02 > 60 < 23 > 60 < 23 > 240 > 240 62.6 61.8

[0355] MF-03 > 60 < 23 > 60 < 23 > 240 > 240 75.5 69.4

[0356] MF-07 43.3 32.0 57.8 24.0 > 240 > 240 75.3 72.5

[0357] 3.7 Pharmacological data

[0358] Further pharmacokinetic data was acquired. The results of the pharmacokinetic experiments in mice are shown for 4 compounds in Figure 8. Depending on the type of infection, different concentrations in plasma, urine and feces are desired. Here, ideal PK data could already be observed for Metro-P3 to treat infections in the gastrointestinal tract: low concentrations of Metro-P3 in plasma and urine, higher and sustained concentration in feces leading to long compound dwell time in gastrointestinal tract high and therefore favourable conditions for e.g. H. pylori treatment in the human stomach. However, compounds MF-01 and MF-03 show higher plasma and urine concentrations without getting cleared rapidly. Thus, the compounds of the present invention can be fine-tuned to different treatment and infection modalities. The features disclosed in the foregoing description, in the claims and / or in the accompanying drawings may, both separately and in any combination thereof, be material for realizing the invention in diverse forms thereof.

[0359] REFERENCES

[0360] Ang CW, Jarrad AM, Cooper MA, Blaskovich MA, Journal of Medicinal Chemistry 2017, 60, 7636-7657.

[0361] Bandow JE, Brbtz H, Leichert LIO, Labischinski H, Hecker M, Antimicrobial Agents and Chemotherapy 2QQ3, 47, 948-955.

[0362] Boucher HW, Talbot GH, Bradley JS, Edwards JE, Gilbert D, Rice LB, Scheid M, Spellberg B, Bartlett J, Clin. Infect. Dis. 2009, 48, 1-12.

[0363] Davies J, Davies D, Microbiology and Molecular Biology Reviews 2010, 74, 417-433.

[0364] Foroumadi A et al. Arch. Pharm. Res. 2004, 27(5), 502-506.

[0365] Jamshidi H et al. Chem. Biol. Drug Des. 2022, 100, 935-946.

[0366] Lehours P, Yilmaz, O., Helicobacter 2007, 12, 1-3.

[0367] Li Z-Z et al. Eur JMedChem. 2019, 179, 723-735.

[0368] Lbfmark S, Edlund C, Nord CE, Clinical infectious diseases 2010, 50, S16-S23.

[0369] Megraud F, Gz / t 2004, 53,1374-1384.

[0370] Megraud F, Gz / t 2007, 56, 1502-1502.

[0371] Mohindra JK, Rauth AM, Cancer Res., 1976, 36, 930-936.

[0372] Perez -Perez GI, Rothenbacher D, Brenner H, Helicobacter 2004, 9, 1-6.

[0373] Vakil N, Megraud F, Gastroenterology 2007, 133, 985-1001.

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Claims

Claims1. A compound having general formula I or II or III(I) (II) (HI) whereinR1is Ci-Ce alkyl or Ci-Ce haloalkyl,R2is Ci-Ce alkyl, C2-Ce alkenyl or C3-Ce alkynyl, wherein for R2said Ci-Ce alkyl, C2-Ce alkenyl or C3-Ce alkynyl is optionally substituted with halogen, OR3or SR3, wherein R3is hydrogen or Ci-Ce alkyl, or a pharmaceutically acceptable salt thereof, under the proviso that for a compound of general formula II or III: when R1is methyl then R2is not ethyl or C1-C4 alkyl substituted with halogen, OH, OMe, NH2, NH(CH3)2or NH(CH2-CH3)2, when R1is CF3then R2is not C1-C4 alkyl substituted with halogen, when R1is isopropyl then R2is not C1-C4 alkyl substituted with halogen; under the proviso that for a compound of general formula III: not both R1and R2are methyl, for use in the treatment of non-viral infections or a disease caused by a non-viral infection.

2. The compound for use according to claim 1, wherein R1is methyl and the compound has formula la or Ila or Illa3. The compound for use according to claim 1 or 2, whereinR2is Ci-Ce alkyl, preferably C1-C4 alkyl, more preferably methyl, ethyl, propyl, isopropyl, n- propyl, n-butyl, isobutyl, sec-butyl, tert-butyl, neopentyl, or n-hexyl; orR2C2-C6 alkenyl, preferably C2-C5 alkenyl, more preferably allyl, vinyl, dimethylallyl, 1- butenyl, 2-butenyl, 3-butenyl, 1- pentenyl, 2-pentenyl, 3-pentenyl, or 4-pentenyl; orR2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, but-l-yne, 1- butynyl, 2-butynyl, 3-butynyl, 1-propynyl, or 2-propynyl.

4. The compound for use according to any one of claims 1 to 3, wherein R2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, which is substituted with halogen, orOR3, wherein R3is hydrogen or methyl, orSR3, wherein R3is hydrogen or methyl.

5. The compound for use according to any one of claims 1 to 4, wherein the compound has formula laalkynyl, preferably C3-C4 alkynyl, more preferably propargyl, or but-l-yne; or R2is Ci-Ce alkyl, preferably methyl; or wherein the compound has formula Ilaand R2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, or but-l-yne; or R2is Ci-Ce alkyl, preferably C1-C4 alkyl, more preferably methyl, or ethyl; R2is C2-C6 alkenyl, preferably C2-C5 alkenyl, more preferably allyl, vinyl, or dimethylallyl; or wherein the compound has formula Illaalkynyl, preferably C3-C4 alkynyl, more preferably propargyl, or but-l-yne.

6. The compound for use according to any one of claims 1 to 5, which is selected from7. The compound for use according to any one of claims 1 to 6, wherein the non-viral infections are infections with protozoa, anaerobic bacteria or microaerophilic bacteria, preferably an infection with Helicobacter pylori, Clostridioides difficile, Trichomonas vaginalis, Giardia intestinalis (G. lambid), Entamoeba histolytica, Fusobacterium nucleatum, or Gardnerella vaginalis, and / or wherein the infection is with a Helicobacter pylori strain having resistance against metronidazole.

8. A compound having general formula IIIwhereinR1is Ci-Ce alkyl or Ci-Ce haloalkyl, andR2is Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl, wherein said Ci-Ce alkyl, C2-C6 alkenyl or C3-C6 alkynyl is optionally substituted with halogen, OR3or SR3, wherein R3is hydrogen or Ci-Ce alkyl, or a pharmaceutically acceptable salt thereof, under the proviso that not both R1and R2are methyl,when R1is methyl then R2is not ethyl or C1-C4 alkyl substituted with halogen, OH,OMe, NH2, or NH(CH3)2, when R1is CF3 then R2is not C1-C4 alkyl substituted with halogen, when R1is isopropyl then R2is not C1-C4 alkyl substituted with halogen.

9. The compound of claim 8, whereinR1is methyl and the compound has formula Illa10. The compound of claim 8 or 9, whereinR2is Ci-Ce alkyl, preferably C1-C4 alkyl, more preferably methyl, ethyl, propyl, isopropyl, n- propyl, n-butyl, isobutyl, sec-butyl, tert-butyl, neopentyl, or n-hexyl; orR2C2-Ce alkenyl, preferably C2-Cs alkenyl, more preferably allyl, vinyl, dimethylallyl, 1- butenyl, 2-butenyl, 3-butenyl, 1- pentenyl, 2-pentenyl, 3-pentenyl, or 4-pentenyl; orR2is C3-C6 alkynyl, preferably C3-C4 alkynyl, more preferably propargyl, but-l-yne, 1- butynyl, 2-butynyl, 3-butynyl, 1-propynyl, or 2-propynyl.

11. The compound of any one of claims 8 to 10, wherein R2is Ci-Ce alkyl, C2-Ce alkenyl or C3-C6 alkynyl, which is substituted with halogen, orOR3, wherein R3is hydrogen or methyl, orSR3, wherein R3is hydrogen or methyl.

12. The compound of any one of claims 8 to 11, wherein the compound has formula Illaalkynyl, preferably C3-C4 alkynyl, more preferably propargyl, or but-l-yne.

13. The compound of any one of claims 8 to 12, which is14. A pharmaceutical composition comprising(i) at least one compound according to any one of claims 8 to 13,(ii) optionally, pharmaceutical excipient(s) and / or carrier.

15. The compound of any one of claims 8 to 13 for use in medicine.

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