4-QUINOLINONE ANTIBACTERIAL COMPOUNDS

Abstract

The present invention relates to the following compounds pounds (I) wherein the integers are as defined in the description, and where the compounds may be useful as medicaments, for instance for use in the treatment of tuberculosis (e.g. in combination).

##STR00001##

Claims

1. A compound of formula (I): ##STR00130## wherein: R.sup.1 is represents is C.sub.1-6 alkyl, —Br, hydrogen or —C(O)N(R.sup.q1)R.sup.q2; R.sup.q1 and R.sup.q2 are, independently, hydrogen or C.sub.1-6 alkyl, or are linked together to form a 3-6 membered carbocyclic ring optionally substituted by one or more C.sub.1-3 alkyl substituents; Sub are one or more optional substituents that are halo, —CN, C.sub.1-6 alkyl, or —O—C.sub.1-6 alkyl (wherein the latter two alkyl moieties are optionally substituted by one or more fluoro atoms); the two “X” rings together area 9-membered bicyclic heteroaryl ring that contains between one and four heteroatoms, and optionally substituted by one or more substituents that are halo and C.sub.1-6 alkyl (itself optionally substituted by one or more fluoro atoms); L.sup.1 is an optional linker group; R.sup.x1 and R.sup.x2 are, independently, hydrogen or C.sub.1-3 alkyl; Z.sup.1 is one of (i) to (vi); ##STR00131## ##STR00132## (v) perfluoro C.sub.1-3 alkyl; or (vi) —F, —Br, —Cl or —CN; ring A is a 5-membered aromatic ring containing at least one heteroatom m), and is optionally substituted by one or more substituents that are, independently, R.sup.f; ring B is a 6-membered aromatic ring containing at least one heteroatom and is optionally substituted by one or more substituents that are, independently, R.sup.g; Y.sup.b is —CH.sub.2 or NH, and R.sup.h is one or more substituents on the 6-membered N and Y.sup.b-containing ring (which R.sup.h substituents may also be present on Y.sup.b); R.sup.a, R.sup.b, R.sup.c, R.sup.d and R.sup.e are, independently, hydrogen or a substituent that is B.sup.1; each R.sup.f, each R.sup.g and each R.sup.h (which are optional substituents), when present, are, independently, a substituent is B.sup.1; each B.sup.1 is independently, a substituent that is: (i) halo; (ii) —R.sup.d1; (iii) —OR.sup.e1; (iv) —C(O)N(R.sup.e2)R.sup.e3 (v) —SF.sub.5; (vi) —N(R.sup.e4)S(O).sub.2R.sup.e5; R.sup.d1 is C.sub.1-6 alkyl optionally substituted by one or more halo atoms; R.sup.e1, R.sup.e2, R.sup.e3, R.sup.e4 and R.sup.e5 are, independently, hydrogen or C.sub.1-6 alkyl optionally substituted by one or more fluoro atoms, or a pharmaceutically-acceptable salt thereof.

2. The compound of claim 1, wherein R.sup.1 is C.sub.1-3 alkyl.

3. The compound of in claim 1, wherein the “X” rings: contain at least one nitrogen atom; and/or contains one, two, three or four heteroatoms in total.

4. The compound of claim 1, wherein the “X” rings together are of formulae (IB): ##STR00133## wherein: one of X.sup.1 and X.sup.2 is N and the other is C; X.sup.3, X.sup.4 and X.sup.5 are C, CH, or a heteroatom; and/or none, one or two of X.sup.3, X.sup.4 and X.sup.5 are a heteroatom and the other is C (or CH) or CH.

5. The compound of claim 1, wherein: L.sup.1 is a direct bond, —O—, —C(R.sup.x1)(R.sup.x2)— or —OCH.sub.2—; R.sup.x1 and R.sup.x2 are, independently, hydrogen.

6. The compound of claim 1, wherein: none, one or two of R.sup.a, R.sup.b, R.sup.c, R.sup.d and R.sup.e is B.sup.1 and the others are hydrogen; and/or one of R.sup.b, R.sup.c and R.sup.d is B.sup.1 and the others are hydrogen.

7. The compound of claim 1, wherein B.sup.1 a substituent that is: (i) fluoro; (ii) —OR.sup.e1; (iii) C.sub.1-3 alkyl substituted by one or more fluoro atom; (iv) —C(O)N(R.sup.e2)R.sup.e3; (v) —N(R.sup.e4)S(O).sub.2R.sup.e5; or (vi) —SF.sub.5.

8. The compound of claim 1, wherein: R.sup.e2 and R.sup.e4 independently are hydrogen; and R.sup.e1, R.sup.e3 and R.sup.e5 are, independently C.sub.1-3 alkyl optionally substituted by one or more fluoro atoms.

9. (canceled)

10. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of the compound of claim 1.

11. (canceled)

12. (canceled)

13. A method of treatment of tuberculosis, comprising administration of a therapeutically effective amount of the compound of claim 1.

14. A combination of (a) the compound of claim 1, and (b) one or more other anti-tuberculosis agent.

15. A product containing (a) a compound of claim 1, and (b) one or more other anti-tuberculosis agent, as a combined preparation for simultaneous, separate or sequential treatment of a bacterial infection.

16. (canceled)

17. (canceled)

18. A method of treatment of tuberculosis, comprising administration of a therapeutically effective amount of a combination of claim 14.

19. A method of enhancing activity of another anti-tuberculosis agent, comprising administering the compound of claim 1 in combination with the another anti-tuberculosis agent.

20. A process for the preparation of a compound of formula (I) of claim 1, comprising: (i) conversion of a compound of formula (II): ##STR00134## by reaction with BBr.sub.3 or NaSCH.sub.3; or (ii) reaction of a compound of formula (III): ##STR00135## with a compound of formula (IV): ##STR00136##

21. The compound of claim 1, wherein: the 9-membered bicyclic heteroaryl ring consists of a 5-membered aromatic ring fused to another 6-membered aromatic ring; and/or the 9-membered bicyclic heteroaryl ring contains between one and four nitrogen, oxygen, or sulfur; and/or L.sup.1 is a direct bond, —O—, —OCH.sub.2—, —C(R.sup.x1)(R.sup.x2)— or —C(O)—N(H)—CH.sub.2—; Z.sup.1 is CF.sub.3; Ring A contains at least one nitrogen atom; Ring B contains at least one nitrogen atom; R.sup.d1 is substituted by one or more fluoro atoms.

22. The compound of claim 2, wherein R.sup.1 is methyl.

23. The compound of claim 3, wherein the “X” rings contain at least one nitrogen atom at the ring junction.

24. The compound of claim 4, wherein the heteroatom of X.sup.3, X.sup.4, and X.sup.5 is N, O, and/or S.

25. The compound of claim 6, wherein: one or two of R.sup.a, R.sup.b, R.sup.c, R.sup.d and R.sup.e is B.sup.1 and the others are hydrogen; or one of R.sup.a, R.sup.b, R.sup.c, R.sup.d and R.sup.e is B.sup.1 and the others are hydrogen; and/or R.sup.c is B.sup.1 and R.sup.b and R.sup.d are hydrogen.

26. The compound of claim 8, wherein R.sup.e1, R.sup.e3, and R.sup.e5 are methyl.

27. The combination of claim 14, wherein the one or more other anti-tuberculosis agent is an inhibitor of the electron transport chain of mycobacteria.

28. The combination of claim 27, wherein the inhibitor of the electron transport chain of mycobacteria is a cytochrome bc inhibitor, an ATP synthase inhibitor, a NDH2 inhibitor and/or an inhibitor of the menaquinone synthesis pathway.

29. The combination of claim 28, wherein the inhibitor of the menaquinone synthesis pathway is a MenG inhibitor.

30. The product of claim 15, wherein the one or more other anti-tuberculosis agent is an inhibitor of the electron transport chain of mycobacteria.

31. The combination of claim 30, wherein the inhibitor of the electron transport chain of mycobacteria is a cytochrome bc inhibitor, an ATP synthase inhibitor, a NDH2 inhibitor and/or an inhibitor of the menaquinone synthesis pathway.

32. The combination of claim 31, wherein the inhibitor of the menaquinone synthesis pathway is a MenG inhibitor.

33. The compound of claim 1, wherein the “X” rings together form: ##STR00137##

Description

PHARMACOLOGICAL EXAMPLES

[0374] In the tests described below, individual compounds of the invention/examples (or combinations containing such compounds, for instance cytochrome bd inhibitors of the invention/examples in combination with one or more other inhibitor(s) of a (different) target of the electron transport chain of mycobacteria, as described herein) may be tested. For instance, in Tests 1 to 4, combinations may be tested (e.g. combinations of test cytochrome bd compounds with known cytochrome bc inhibitors, such as Q203 and Compound X). Where a control cytochrome bd compound is employed, then CK-2-63 is employed.

[0375] The compound Q203 (cytochrome bc1 inhibitor) may be prepared in accordance with the procedures in J. Medicinal Chemistry, 2014, 57 (12), pp 5293-5305, as well as, in WO 2011/113606 (see Compound 289 “6-chloro-2-ethyl-N-(4-(4-(4-(trifluoromethoxy)phenyl)piperidin-1-yl)benzyl)imidazo[1,2-a]pyridine-3-carboxamide”).

[0376] Compound X is 6-chloro-2-ethyl -N-({4-[2-(trifluoromethanesulfonyl)-2-azaspiro[3.3]heptan-6-yl]phenyl}methyl)imidazo[1,2-a]pyridine-3-carboxamide, which is described as Compound 154 of WO 2017/001660 and may be prepared according to the procedures described therein.

[0377] CK-2-63 may be prepared in accordance with the procedures disclosed in WO 2017/103615 (see experimental and the disclosures therein, referring to WO 2012/2069856, where an experimental procedure is provided for “3-methyl-2-(4-(4-(trifluoromethoxy)phenoxy)phenyl)quinolin-4(1H)-one”).

[0378] MIC Determination Against M. tuberculosis: Test 1

[0379] Test compounds and reference compounds were dissolved in DMSO and 1 μl of solution was spotted per well in 96 well plates at 200× the final concentration. Column 1 and column 12 were left compound-free, and from column 2 to 11 compound concentration was diluted 3-fold. Frozen stocks of Mycobacterium tuberculosis strain EH4.0 expressing green-fluorescent protein (GFP) were previously prepared and titrated. To prepare the inoculum, 1 vial of frozen bacterial stock was thawed to room temperature and diluted to 5×10 exp5 colony forming units per ml in 7H9 broth. 200 μl of inoculum, which corresponds to 1×10 exp5 colony forming units, were transferred per well to the whole plate, except column 12. 200 μl 7H9 broth were transferred to wells of column 12. Plates were incubated at 37° C. in plastic bags to prevent evaporation. After 7 days, fluorescence was measured on a Gemini EM Microplate Reader with 485 excitation and 538 nm emission wavelengths and IC.sub.50 and/or pIC.sub.50 values (or the like, e.g. IC.sub.50, IC.sub.90, pIC.sub.90, etc) were (or may be) calculated.

[0380] MIC Determination Against M. tuberculosis: Test 2

[0381] Appropriate solutions of experimental and reference compounds were made in 96 well plates with 7H9 medium. Samples of Mycobacterium tuberculosis strain H37Rv were taken from cultures in logarithmic growth phase. These were first diluted to obtain an optical density of 0.3 at 600 nm wavelength and then diluted 1/100, resulting in an inoculum of approximately 5×10 exp5 colony forming units per ml. 100 μl of inoculum, which corresponds to 5×10 exp4 colony forming units, were transferred per well to the whole plate, except column 12. Plates were incubated at 37° C. in plastic bags to prevent evaporation. After 7 days, resazurin was added to all wells. Two days later, fluorescence was measured on a Gemini EM Microplate Reader with 543 excitation and 590 nm emission wavelengths and MIC.sub.50 and/or pIC.sub.50 values (or the like, e.g. IC.sub.50, IC.sub.90, pIC.sub.90, etc) were (or may be) calculated.

[0382] Time Kill Kinetics Assays: Test 3

[0383] Bactericidal or bacteriostatic activity of the compounds can be determined in a time kill kinetic assay using the broth dilution method. In this assay, the starting inoculum of M. tuberculosis (strain H37Rv and H37Ra) is 10.sup.6 CFU/ml in Middlebrook (1×) 7H9 broth. The test compounds (cyt bd inhibitors) are tested in combination with a cyt bc inhibitor (for example Q203 or Compound X) at the concentration ranging from 10-30 μM to 0.9-0.3 μM respectively. Tubes receiving no antibacterial agent constitute the culture growth control. The tubes containing the microorganism and the test compounds are incubated at 37° C. After 0, 1, 4, 7, 14 and 21 days of incubation samples are removed for determination of viable counts by serial dilution (10.sup.0 to 10.sup.−6) in Middlebrook 7H9 medium and plating (100 μl) on Middlebrook 7H11 agar. The plates are incubated at 37° C. for 21 days and the number of colonies are determined. Killing curves can be constructed by plotting the log.sub.10 CFU per ml versus time. A bactericidal effect of a cytochrome be and cytochrome bd inhibitor (either alone or in combination) is commonly defined as 2-log.sub.10 decrease (decrease in CFU per ml) compared to Day 0. The potential carryover effect of the drugs is limited by using 0.4% charcoal in the agar plates, and by serial dilutions and counting the colonies at highest dilution possible used for plating.

[0384] Phenotypic Assay to Determine the O.sub.2 Consumption Rate of Mycobacterium tuberculosis: Test 4

[0385] The aim of this assay is to evaluate the O.sub.2 consumption rate of Mycobacterium tuberculosis (Mtb) bacilli after inhibition of cyt bc1 and cyt bd, using extracellular flux technology. Inhibition of cyt bc1 (e.g. using known inhibitors such as Q203 or Compound X) forces the bacillus to use the less energetically efficient terminal oxidase cyt bd. The inhibition of cyt bd will cause a significant decrease O.sub.2 consumption. A sustained decrease of O.sub.2 consumption under membrane potential disrupting conditions, via the addition of the uncoupler CCCP, will show to the efficacy of the cyt bd inhibitor. The oxygen consumption rate (OCR) of Mtb (stain H37Ra) bacilli adhered to the bottom of a Cell-Tak (BD Biosciences) coated XF cell culture microplate (Agilent), at 5×10.sup.6 bacilli per well, was measured using the Agilent Seahorse XFe96. Prior to the assay Mtb bacilli are cultured for two days to an OD.sub.600 ˜0.7-0.9 in liquid medium, using 7H9 supplemented with 10% and 0.02% Tyloxapol. The assay media used is unbuffered 7H9 only supplemented with 0.2% glucose. For this assay the Compound X (final concentration of 0.9 μM, Compound X), is used to inhibit cyt bc1 and the cyt bd inhibitor, CK-2-63 (final concentration of 10 μM), is used as a positive control. The uncoupler CCCP is used at a final concentration of 1 μM.

[0386] In general, four basal OCR measurements are taken before the automatic addition of Compound X, through drug port A of the sensor cartridge, after which seven more OCR measurements are taken to allow enough time for the inhibition of cyt bc1. Next the cyt bd test compounds (final concentration of 10 μM), as well as the positive and negative controls (assay media with a final DMSO concentration of 0.4%), are added (drug port B) followed by seven OCR measurements. Finally, CCCP is added followed by three OCR measurements, this is done twice (drug ports C and D). For the control's measurements are performed in eight replicate wells and for the assay compounds six replicate wells per condition. Compounds are scored for their sustained inhibition of cyt bd in relation to the positive and negative controls.

[0387] Further Phenotypic Assay: Using a Cytochrome bc Knock-Out TB Strain and MIC Determination Against M. tuberculosis: Test 5

[0388] Appropriate solutions of experimental and reference compounds were made in 384 well plates with 7H9 medium. Samples of Mycobacterium tuberculosis strain H37Rv ΔctaE-ΔqcrCAB (Nat Commun 10, 4970, 2019, https://doi.org/10.1038/s41467-019-12956-2) were taken from cultures in logarithmic growth phase. These were first diluted to obtain an optical density of 0.4 at 600 nm wavelength and then diluted 1/150, resulting in an inoculum of approximately 5×10 exp5 colony forming units per ml. 30 μl of inoculum, which corresponds to 5×10 exp5 colony forming units, were transferred per well to the whole plate, except columns 23-24. Plates were incubated at 37° C., in an extra humidified incubator, in plastic bags to prevent evaporation. After 10 days, optical density at 620 nm wavelength was measured on an EnVision 2105 Multimode Plate Reader with a Photometric 620/8 excitation filter, and MIC.sub.50 and/or pIC.sub.50 values (or the like, e.g. IC.sub.50, IC.sub.90, pIC.sub.90, etc) were (or may be) calculated.

[0389] Pharmacological Results

[0390] Biological Data—Example A

[0391] Compounds of the invention/examples (or combinations, e.g. compounds of the invention/examples in combination with one or more other inhibitors of a target of the electron transport chain), for example when tested in any of Tests 1 to 3, may display activity.

[0392] Biological Data—Example B

[0393] Compounds of the examples were tested in Test 4 described above (in section “Pharmacological Examples”; O.sub.2 consumption rate testing), together with Compound X—a known cytochrome bc inhibitor—as described above, and the following results were obtained:

TABLE-US-00003 (i) % OCR after cyt (ii) % OCR after Example bd inhibitor cccp 1 26.9 35.1 2 24.8 38.3 3 28.75 38.75 4 32.75 40.05 5 43.48 49.26 7 36.5 54.6 6 39.7 58.4 8 82.67 59.43 9 58.71 62.22 10 43.4 63.9 11 62.79 69.17 12 62.09 79.37 13 61.74 83.9 15 91.58 94.75 16 80.34 95.29 17 98.64 97.68 18 76.01 98.7 19 99.4 100.5 20 78.5 102.86 21 81.6 103.4 22 82.7 109.8 23 97.51 110.11 24 99.13 113.29 25 65.2 114.3 26 71 116.7 27 111.36 117.59 28 123.3 117.7 29 64.6 118.5 30 93.8 120.2 31 68.97 120.39 33 61.2 123.6 35 102.9 129.2 36 88.6 130.6 37 93.8 132.68 38 95.63 132.88 39 110.6 133.8 40 95.33 134.33 41 84.09 137.15 42 80.1 137.9 43 109.2 138 44 76 138.6 45 94.41 138.84 46 91.4 140.4 47 99.34 141.23 49 101.3 143.58 50 114.33 144.07 52 82.2 148.1 55 109.2 148.3 57 102 149.2 58 139.74 153.33 59 107.31 154.34 60 114.505 155.065 61 108 155.2 63 85.8 157.9 64 109.65 158.95 65 99.86 158.97 67 99.28 161.97 68 129.7 164.4 69 150.98 165.1 70 102.2 165.65 71 95.71 167.55 72 99.84 169.69 73 105.2 171.2 74 86.4 173.3 76 111.72 175.05 77 110.77 175.26 78 135.74 178.3 79 112 180.5 80 122.15 180.98 81 98.58 182.7 82 96.77 186.45 86 32,57 39,44 87 33,23 38,98 90 25,77 30,69 110 40,37 51,51 124 31,72 37,02 125 45,66 64,44 126 43,71 51,74 127 49,62 65,3  88  63,905 76 89 55,82 62,29 91 49,11 56,23 98 54,17 73,86 107 58,92 76,98 112 31,36 38,59 116 72,01 85,87 120 59,83 89,24 123 50,75  60,795

[0394] Biological Data—Example C

[0395] Compounds of the examples are/were tested in Test 3 (the kill kinetics) described above, obtaining results expressed as a log reduction in CFUs per ml as compared to Day 0. The following results were obtained.

TABLE-US-00004 Log Day 21- Log Day 0 Log Day 21 Log Day 0 Control 6.66 9.16 +2.50 Compound X (0.17 μg/ml) 6.66 5.93 −0.73 Compound 6 (12 μg/ml) 6.66 9.06 +2.40 Compound 7 (12 μg/ml) 6.66 9.13 +2.47 Compound X (0.17 μg/ml) + 6.66 1.40 −5.26 Compound 6 (12 μg/ml) Compound X (0.17 μg/ml) + 6.66 2.27 −4.39 compound 6 (1.2 μg/ml) Compound X (0.17 μg/ml) + 6.66 5.76 −0.89 compound 6 (0.12 μg/ml) Compound X (0.17 μg/ml) + 6.66 1.40 −5.26 Compound 7 (12 μg/ml) Compound X (0.17 μg/ml) + 6.66 1.30 −5.36 Compound 7 (1.2 μg/ml) Compound X (0.17 μg/ml) + 6.66 5.39 −1.27 Compound 7 (0.12 μg/ml)

TABLE-US-00005 Log Day 0 Log Day 21 Log Day 21-Log Day 0 Control 5.56 8.73 +3.17 Q203 (0.168 μg/ml) 5.56 2.59 −2.97 Compound 6 (12 μg/ml) 5.56 8.64 +3.08 Compound 2 (12 μg/ml) 5.56 8.69 +3.13 Q203 (0.168 μg/ml) + 5.56 1.00 −4.56 compound 6 (12 μg/ml) Q203 (0.168 μg/ml) + 5.56 1.00 −4.56 compound 2 (12 μg/ml)

[0396] Biological Data—Example D

[0397] Compounds of the examples were re-tested in Test 5 described above, and the following results were obtained:

TABLE-US-00006 Compound number pIC.sub.50 35 5.622 23 <4.000 42 4.513 44 5.011 63 4.118 52 <4.000 29 4.017 22 5.426 25 5.510 6 6.185 2 5.876 55 4.443 29 5.023 4 5.904 33 <4.000 39 <4.000 46 <4.301 73 <4.000 30 5.535 28 4.998 1 5.787 21 4.945 70 <4.301 57 <4.301 12 6.053 68 <4.000 43 <4.000 74 <4.000 7 5.851 26 5.114 18 5.726 79 <4.000 5 5.896 54 5.226 72 6.584 20 5.663 40 <4.000 65 <5.000 41 4.470 77 5.734 80 <4.602 67 4.989 81 4.778 45 5.225 11 5.585 38 5.312 76 <5.301 71 <5.000 82 <5.301 31 <4.301 49 <4.301 58 <4.602 17 <5.301 15 4.244 27 <4.301 9 5.884 8 5.432 24 5.530 16 5.932 60 <4.301 13 5.385 86 5.881 87 6.261 88 5.272 89 6.241 90 5.828 91 5.760 98 5.704 107 5.479 110 5.423 112 5.732 116 5.338 120 6.157 123 5.383 124 5.771 125 6.178 126 4.965 127 5.491

[0398] Further Data

[0399] The compounds of the invention/examples may have advantages associated with in vitro potency, kill kinetics (i.e. bactericidal effect) in vitro, PK properties, food effect, safety/toxicity (including liver toxicity, coagulation, 5-LO oxygenase), metabolic stability, Ames II negativity, MNT negativity, aqueous based solubility (and ability to formulate) and/or cardiovascular effect e.g. on animals (e.g. anesthetized guinea pig). The data below that was generated/calculated may be obtained using standard methods/assays, for instance that are available in the literature or which may be performed by a supplier (e.g. Microsomal Stability Assay—Cyprotex, Mitochondrial toxicity (Glu/Gal) assay—Cyprotex, as well as literature CYP cocktail inhibition assays).

TABLE-US-00007 Mitotoxicity data: cpd Δ IC50,glu/ number IC50, glu IC50, gal IC50,gal Score 1 [x]100 [x]19.8 [x]5.06 positive 2 [x]100 [x]100 [x]0 negative 4 [x]50 [x]50 [x]0 negative 5 [x]106.39 [x]45.87 [x]2.32 inconclusive 7 [x]20 [x]20 [x]0 negative 6 26.5 22.1 1.2 negative 6 (repeat) 15.6 21.9 1.4 negative 8 [x]163.33 [x]180 [x]0.91 negative 12 [x]100 [x]100 [x]0 negative 13 [x]200 [x] 141.1 [x]1.42 negative 15 [x]44.26 [x]200 [x]0.22 negative 16 [x]200 [x]200 [x]0 negative 18 [x]23.35 [x]15.75 [x]1.48 negative 19 [x]100 [x]100 [x]0 negative 20 [x] 176.1 [x]125.31 [x]1.41 negative 21 [x]50 [x]50 [x]0 negative 22 139.5 9.9 >13.3 positive 23 200 200 n.a. negative 24 [x]20 [x]20 [x]0 negative 25 200 9.9 >20.2 positive 26 [x]13.31 [x]9.59 [x]1.39 negative 27 [x]100 [x]100 [x]0 negative 28 [x]100 [x]58.7 [x]1.7 negative 29 79.7 53.7 1.5 negative 30 [x]100 [x]100 [x]0 negative 31 [x]200 [x]75.95 [x]2.63 inconclusive 33 [x]100 [x]100 [x]0 negative 39 [x]100 [x]100 [x]0 negative 40 [x]200 [x]200 [x]0 negative 41 [x]53.91 [x]54.72 [x]0.99 negative 42 200 200 n.a. negative 43 [x]26.3 [x]10.6 [x]2.48 inconclusive 44 47.8 58.5 0.8 negative 45 [x]20 [x]20 [x]0 negative 46 [x]100 [x]100 [x]0 negative 50 [x]100 [x]86.53 [x]1.16 negative 52 133.5 108.0 1.2 negative 55 [x]200;[x]100 [x]200;[x]100 [x]0;[x]0 negative 57 [x]100 [x]100 [x]0 negative 58 [x]100 [x]100 [x]0 negative 60 [x]100 [x]100 [x]0 negative 63 200 96.8 >1.7 inconclusive (precipitation) 65 [x]20 [x]20 [x]0 negative 67 [x]30.03 [x]0.39 [x]76.87 positive 68 [x]100 [x]100 [x]0 negative 70 [x]100 [x]27.1 [x]3.69 inconclusive 72 [x]100 [x]100 [x]0 negative 73 [x]100 [x]100 [x]0 negative 74 [x]134.52 [x]121.15 [x]l.ll negative 77 [x]200 [x]200 [x]0 negative 79 [x]100 [x]100 [x]0 negative 80 [x]50 [x]50 [x]0 negative 81 [x]200 [x]200 [x]0 negative 82 [x]50 [x]50 [x]0 negative 86 [x]50 [x]50 [x]0 negative 87 [x]6.2 [x]5.16 [x]1.21 inconclusive 90 [x]200 [x]200 [x]0 negative 110 [x]200 [x]200 [x]0 negative 124 [x]200 [x]200 [x]0 negative 125 [x]200 [x]200 [x]0 negative 126 [x]25 [x]15.5 [x]1.61 inconclusive 127 [x]18.1 [x]15.85 [x]1.14 negative

[0400] In the table above, “negative” means that in the test, it was found to have low mitotoxicity (and hence no mitotoxicity alerts), “positive” means that there were some mitotoxicity alerts and “inconclusive” means that no accurate conclusion could be drawn, e.g. due to issues with the compound being tested in the assay, e.g. solubility or precipitation issues (e.g. compound may not be soluble enough or may precipitate).

[0401] In view of the data above, compounds of the invention/examples may be found to be advantageous as no mitotoxicity alerts were observed (e.g. in the Glu/Gal assay).

[0402] The following data were also generated:

[0403] Compound 6:

[0404] CVS—rCaCh, rNaCh & hERG IC.sub.50 (μm)=>10/>10/>10

[0405] AMES II b (+/−rat S9)=negative

[0406] GSH and CN adducts=negative

[0407] PK parameters in mice T.sub.1/2(h), CI (mL/min/kg), Fab %=5.6/1.69/64

[0408] CTCM Ca.sup.2+ transient h-cardiomyocytes HTS (μm)=0.1 μm, 0.2 μm, 0.5 μm, 1 μm, 2.5 μm, 5 μm (all no)

TABLE-US-00008 Cardio tox rCaCH (plC50), rNaCH (IC50), Compound hERG(DOF) (IC50) 44 3.8, 1.2, 1.5 63 >3.0, 2.8, >10 52 ≈1.9, 7.4, >10 29 3.3, 2.3, ≈9.1 22 >10, 6.9, >10 25 >10, >10, >10 55 >10, >10, >10 19 >10, >10, >10 4 >10, 2.3, >10 33 >10, >10, >10 70 >10, >10, >10 12 >10, >10, >3.02 26 >10, >10, >10 18 >10, 1.51, >10 5 >10, >10, >10 58 >10, >10, >10

[0409] Compounds of the invention/examples, may therefore have the advantage that: [0410] No in vitro cardiotoxicity is observed (for example either due to the CVS results or due to the Glu/Gal assay results, for instance low mitotoxicity (<3 in the Glu/Gal assay indicates no mitotoxicity alerts); and/or [0411] No reactive metabolite formation is observed (e.g. GSH);

[0412] for instance as compared to other compounds, for instance prior art compounds.