Antibacterial compounds

Abstract

The present invention relates to the following compounds for use in the treatment of a bacterial infection ##STR00001##
wherein the integers are as defined in the description. The invention also relates to compounds for use as medicaments, pharmaceutical compositions and some novel compounds.

Claims

1. A compound selected from the group consisting of: ##STR00021## and pharmaceutically acceptable salts thereof.

2. A pharmaceutical composition comprising a compound as claimed in claim 1 and a pharmaceutically acceptable carrier.

Description

EXPERIMENTAL PART

Preparation of Compound 1

(1) ##STR00014##

(2) A solution of 2-Amino-4,6-difluoro-1,3-benzothiazole (119256-40-5, 0.22 g, 1.18 mmol), 1-Adamantyl isocyanate (0.42 g, 2.36 mmol) and triethylamine (0.27 mL, 1.97 mmol) in THF (4 mL) was stirred and heated overnight at 60° C. The solution was cooled down to room temperature. Water and DCM were added. The organic layer was separated, dried over MgSO.sub.4, filtered and evaporated. The residue was purified by preparative LC on (dry loading 25 g+5 g 15-40 μm merck). Mobile phase (Gradient from 90% HEPTANE, 10% AcOEt to 70% HEPTANE, 30% AcOEt). Pure fractions were collected and evaporated to give a white powder, 0.125 g. This compound was then purified by achiral SFC on (DIETHYLAMINOPROPYL 5 μm 150×21.2 mm). Mobile phase (75% CO2, 25% MeOH). Pure fractions were collected and evaporated to a white powder, 0.09 g.

(3) The residue was crystallized from DIPE, filtered off and dried under vacuum at 60° C. to give Compound 1 as a white powder, 0.084 g, 20%, m.p.>260° C.

(4) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 10.67 (br. s., 1H), 7.70 (dd, J=1.5, 8.1 Hz, 1H), 7.22-7.31 (m, 1H), 7.05 (d, J=8.1 Hz, 1H), 3.84 (d, J=8.1 Hz, 1H), 1.57-1.90 (m, 14H)

Preparation of Compound 2

(5) ##STR00015##

(6) Compound 2 was prepared in the same way as Compound 1 from 2-amino-5-chlorobenzoxazole (61-80-3, 0.2 g, 1.19 mmol) affording the expected Compound 2, 0.161 g, 39%, m.p.>250° C.

(7) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.98 (br. s., 1H), 8.16 (br. s., 1H), 7.48-7.61 (m, 2H), 7.23 (dd, J=2.1, 8.7 Hz, 1H), 1.95-2.15 (m, 9H), 1.66 (br. s., 6H)

Preparation of Compound 3

(8) ##STR00016##

(9) A solution of 2-Amino-4,6-difluoro-1,3-benzothiazole (3 g, 16.11 mmol) and 1,1′-carbonyldiimidazole (2.87 g, 17.72 mmol) in dichloromethane (60 mL) was stirred overnight at room temperature. The precipitate was filtered off, washed with EtOH and dried under vacuum at 60° C. affording intermediate A as a white powder, 2.49 g, 55%, used as such for next step.

(10) A solution of intermediate A (1.99 g, 7.1 mmol), 2-Aminoadamantane hydrochloride (1.47 g, 7.81 mmol) and triethylamine (1.57 mL, 11.36 mmol) in THF (20 mL) was stirred at 60° C. overnight. The solution was cooled down to room temperature. Water and DCM were added. The organic layer was separated, dried over MgSO.sub.4, filtered off and evaporated. The residue was purified by preparative LC (Stationary phase: irregular SiOH 15-40 μm 300 g MERCK), Mobile phase: 80% HEPTANE, 20% AcOEt). Pure fractions were collected and the solvent was evaporated to give a white powder, 0.33 g. The compound was crystallized from DIPE, filtered off and dried under vacuum at 60° C. affording Compound 3 as a white powder, 0.271 g, 10%, m.p.=272° C.

(11) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.56 (br. s., 1H), 7.68 (dd, J=1.6, 8.2 Hz, 1H), 7.21-7.33 (m, 1H), 6.47 (s, 1H), 2.05 (br. s., 3H), 1.95 (br. s., 6H), 1.64 (br. s., 6H)

Preparation of Compound 4

(12) ##STR00017##

(13) A solution of 2-amino-5-chlorobenzoxazole (61-80-3, 0.3 g, 1.78 mmol) and 1,1′-carbonyldiimidazole (0.32 g, 1.96 mmol) in dichloromethane (6 mL) was stirred overnight at room temperature. The precipitate was filtered off, washed with EtOH and dried under vacuum at 60° C. affording intermediate A as a white powder, 0.19 g, 40%, used as such for next step.

(14) A solution of intermediate A (0.19 g, 0.72 mmol), 2-Aminoadamantane hydrochloride (0.15 g, 0.79 mmol) and triethylamine (0.16 mL, 1.15 mmol) in THF (4 mL) was stirred at 60° C. overnight. The solution was cooled down to room temperature. Water and DCM were added. The organic layer was separated, dried over MgSO.sub.4, filtered off and evaporated. Purification was carried out by flash chromatography over silica gel (40 g, 15-40 μm, Heptane/EtOAc from 90/10 to 70/30). Pure fractions were collected and the solvent was removed. The residue was crystallized from DIPE, filtered off and dried under vacuum at 60° C. affording Compound 4 as a white powder, 0.081 g, 33%, m.p.>250° C.

(15) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 11.24 (br. s., 1H), 8.81 (d, J=7.6 Hz, 1H), 7.52-7.63 (m, 2H), 7.25 (dd, J=2.1, 8.7 Hz, 1H), 3.91 (d, J=7.6 Hz, 1H), 1.58-1.99 (m, 14H)

Preparation of Compound 5

(16) ##STR00018##

(17) A solution of 2-Amino-6-(trifluoromethyl)-benzothiazole (777-12-8, 0.3 g, 1.39 mmol) and 1,1′-carbonyldiimidazole (0.25 g, 1.53 mmol) in dichloromethane (6 mL) was stirred overnight at room temperature. The precipitate was filtered off, washed with EtOH and dried under vacuum at 60° C. affording intermediate A as a white powder, 0.231 g, 53%, used as such for next step.

(18) A solution of intermediate A (0.231 g, 0.74 mmol), 2-Aminoadamantane hydrochloride (0.15 g, 0.81 mmol) and triethylamine (0.16 mL, 1.18 mmol) in THF (8 mL) was stirred at 60° C. overnight. The solution was cooled down to room temperature. Water and DCM were added. The organic layer was separated, dried over MgSO.sub.4, filtered off and evaporated. Purification was carried out by flash chromatography over silica gel (40 g, 15-40 μm, Heptane/EtOAc from 80/20 to 60/40). Pure fractions were collected and the solvent was removed. The residue was crystallized from DIPE, filtered off and dried under vacuum at 60° C. affording Compound 5 as a white powder, 0.141 g, 48%, m.p.>250° C.

(19) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.69 (br. s., 1H), 8.39 (s, 1H), 7.77 (d, J=8.5 Hz, 1H), 7.66 (dd, J=1.6, 8.5 Hz, 1H), 7.12 (d, J=7.9 Hz, 1H), 3.84 (d, J=7.9 Hz, 1H), 1.69-1.91 (m, 13H), 1.55-1.64 (m, 1H)

Preparation of Compound 6

(20) ##STR00019##

(21) A solution of 2-Amino-5,6-dimethyl-benzothiazole (29927-08-0, 0.25 g, 1.39 mmol) and 1,1′-carbonyldiimidazole (0.25 g, 1.53 mmol) in dichloromethane (6 mL) was stirred overnight at room temperature. The precipitate was filtered off, washed with EtOH and dried under vacuum at 60° C. affording intermediate A as a white powder, 0.351 g, 93%, used as such for next step.

(22) A solution of intermediate A (0.351 g, 1.29 mmol), 2-Aminoadamantane hydrochloride (0.27 g, 1.42 mmol) and triethylamine (0.29 mL, 2.06 mmol) in THF (8 mL) was stirred at 60° C. overnight. The solution was cooled down to room temperature. Water and DCM were added. The organic layer was separated, dried over MgSO.sub.4, filtered off and evaporated. Purification was carried out by flash chromatography over silica gel (40 g, 15-40 μm, Heptane/EtOAc from 90/10 to 70/30). Pure fractions were collected and the solvent was removed. The residue was crystallized from DIPE, filtered off and dried under vacuum at 60° C. affording Compound 6 as a white powder, 0.038 g, 8%, m.p.>260° C.

(23) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.36 (br. s., 1H), 7.60 (s, 1H), 7.40 (s, 1H), 7.17 (br. s., 1H), 3.82 (d, J=7.2 Hz, 1H), 2.28 (d, J=4.7 Hz, 6H), 1.54-1.89 (m, 14H)

Preparation of Compound 7

(24) ##STR00020##

(25) A solution of 2-Amino-benzothiazole-6-carboxylic acid methyl ester (0.3 g, 1.46 mmol) and 1,1′-carbonyldiimidazole (0.26 g, 1.6 mmol) in dichloromethane (6 mL) was stirred overnight at room temperature. The precipitate was filtered off, washed with EtOH and dried under vacuum at 60° C. affording intermediate A as a white powder, 0.426 g, 97%, used as such for next step.

(26) A solution of intermediate A (0.426 g, 1.41 mmol), 2-Aminoadamantane hydrochloride (0.29 g, 1.55 mmol) and triethylamine (0.31 mL, 2.25 mmol) in THF (8 mL) was stirred at 60° C. overnight. The solution was cooled down to room temperature. Water and CH.sub.2Cl.sub.2 were added. The organic layer was separated, dried over MgSO.sub.4, filtered off and evaporated. The residue was purified by achiral SFC (Stationary phase: DIETHYLAMINOPROPYL 5 μm 150×21.2 mm), Mobile phase: 85% CO2, 15% MeOH). Pure fractions were collected and the solvent was evaporated to give intermediate B as a white powder, 0.23 g, 42%.

(27) Lithium hydroxide monohydrate (0.22 g, 2.88 mmol) was added portion wise to a solution of intermediate B (0.222 g, 0.58 mmol) in THF (3 mL) and water (0.3 mL). The solution was stirred and heated at 60° C. for 2 hours. THF was evaporated and the mixture was acidified with HCl 3N. AcoEt was added, and the organic layer was separated, dried over MgSO.sub.4, filtered and evaporated to give 0.085 g, 40%. A solution of this intermediate (0.085 g, 0.23 mmol), Dimethylamine hydrochloride (0.028 g, 0.34 mmol), 1-hydroxybenzotriazole (0.037 g, 0.27 mmol), 1-(3-Dimethylaminopropoyl)-3-ethylcarbodiimide hydrochloride (0.053 g, 0.27 mmol) and N,N-diisopropylethylamine (0.082 mL, 0.46 mmol) in CH.sub.2Cl.sub.2 (2 mL) was stirred overnight at room temperature. Water and CH.sub.2Cl.sub.2 were added. The organic layer was extracted, washed twice with brine, dried over MgSO.sub.4, filtered and evaporated.

(28) Purification was carried out by flash chromatography over silicagel (15-40 μm, 24 g, CMA from 100/0/0 to 97/3/0.1). Pure fractions were collected and the solvent was evaporated to give Compound 7 as a white powder, 0.036 g, 39%, m.p.=224° C.

(29) .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 10.56 (br. s., 1H), 7.97 (s, 1H), 7.62 (d, J=8.2 Hz, 1H), 7.39 (dd, J=1.4, 8.2 Hz, 1H), 7.14 (d, J=5.7 Hz, 1H), 3.84 (d, J=5.7 Hz, 1H), 2.97 (br. s., 6H), 1.54-1.91 (m, 14H)

(30) Analytical Methods

(31) LCMS

(32) The mass of some compounds was recorded with LCMS (liquid chromatography mass spectrometry). The methods used are described below.

(33) General Procedure A

(34) The HPLC measurement was performed using an Alliance HT 2795 (Waters) system comprising a quaternary pump with degasser, an autosampler, a diode-array detector (DAD) and a column as specified in the respective methods below, the column is hold at a temperature of 30° C. Flow from the column was split to a MS spectrometer. The MS detector was configured with an electrospray ionization source. The capillary needle voltage was 3.15 kV and the source temperature was maintained at 110° C. on the ZQ™ (simple quadrupole Zspray™ mass spectrometer from Waters). Nitrogen was used as the nebulizer gas. Data acquisition was performed with a Waters-Micromass MassLynx-Openlynx data system.

(35) General Procedure B

(36) The LC measurement was performed using an Acquity UPLC (Waters) system comprising a binary pump, a sample organizer, a column heater (set at 55° C.), a diode-array detector (DAD) and a column as specified in the respective methods below. Flow from the column was split to a MS spectrometer. The MS detector was configured with an electrospray ionization source. Mass spectra were acquired by scanning from 100 to 1000 in 0.18 seconds using a dwell time of 0.02 seconds. The capillary needle voltage was 3.5 kV and the source temperature was maintained at 140° C. Nitrogen was used as the nebulizer gas. Data acquisition was performed with a Waters-Micromass MassLynx-Openlynx data system.

(37) General procedure C

(38) The HPLC measurement was performed using an Agilent 1100 series liquid chromatography system comprising a binary pump with degasser, an autosampler, a column oven, a UV detector and a column as specified in the respective methods below. Flow from the column was split to a MS spectrometer. The MS detector was configured with an electrospray ionization source. The capillary voltage was 3 kV, the quadrupole temperature was maintained at 100° C. and the desolvation temperature was 300° C. Nitrogen was used as the nebulizer gas. Data acquisition was performed with an Agilent Chemstation data system.

(39) General Procedure D

(40) The LC measurement was performed using a UPLC (Ultra Performance Liquid Chromatography) Acquity (Waters) system comprising a binary pump with degasser, an autosampler, a diode-array detector (DAD) and a column as specified in the respective methods below, the column is hold at a temperature of 40° C. Flow from the column was brought to a MS detector. The MS detector was configured with an electrospray ionization source. The capillary needle voltage was 3 kV and the source temperature was maintained at 130° C. on the Quattro (triple quadrupole mass spectrometer from Waters). Nitrogen was used as the nebulizer gas. Data acquisition was performed with a Waters-Micromass MassLynx-Openlynx data system.

(41) Method 1

(42) In addition to general procedure A: Reversed phase HPLC was carried out on a Sunfire C18 column (3.5 μm, 4.6×100 mm) with an initial flow rate of 0.8 ml/min. Two mobile phases (mobile phase A: 35% 6.5 mM ammonium acetate+30% acetonitrile+35% formic acid (2 ml/1); mobile phase B: 100% acetonitrile) were employed to run a gradient condition from 100% A (hold for 1 minute) to 100% B in 4 minutes, hold at 100% B at a flow rate of 1.2 ml/min for 4 minutes and reequilibrated with initial conditions for 3 minutes. An injection volume of 10 μl was used. Cone voltage was 20 V for positive and negative ionization mode. Mass spectra were acquired by scanning from 100 to 1000 in 0.4 seconds using an interscan delay of 0.3 seconds.

(43) Method 2

(44) In addition to general procedure A: Reversed phase HPLC was carried out on a Sunfire C18 column (3.5 μm, 4.6×100 mm) with an initial flow rate of 0.8 ml/min. Two mobile phases (mobile phase A: 25% 7 mM ammonium acetate+50% acetonitrile+25% formic acid (2 ml/l); mobile phase B: 100% acetonitrile) were employed to run a gradient condition from 100% A (hold for 1 minute) to 100% B in 4 minutes, hold at 100% B at a flow rate of 1.2 ml/min for 4 minutes and reequilibrated with initial conditions for 3 minutes). An injection volume of 10 μl was used. Cone voltage was 20 V for positive and negative ionization mode. Mass spectra were acquired by scanning from 100 to 1000 in 0.4 seconds using an interscan delay of 0.3 seconds.

(45) Method 3

(46) In addition to general procedure B: Reversed phase UPLC (Ultra Performance Liquid Chromatography) was carried out on a bridged ethylsiloxane/silica hybrid (BEH) C18 column (1.7 μm, 2.1×50 mm; Waters Acquity) with a flow rate of 0.8 ml/min. Two mobile phases (mobile phase A: 0.1% formic acid in H.sub.2O/methanol 95/5; mobile phase B: methanol) were used to run a gradient condition from 95% A and 5% B to 5% A and 95% B in 1.3 minutes and hold for 0.2 minutes. An injection volume of 0.5 μl was used. Cone voltage was 10 V for positive ionization mode and 20 V for negative ionization mode.

(47) Method 4

(48) In addition to general procedure C: Reversed phase HPLC was carried out on a YMC-Pack ODS-AQ C18 column (4.6×50 mm) with a flow rate of 2.6 ml/min. A gradient run was used from 95% water and 5% acetonitrile to 95% acetonitrile in 7.30 minutes and was hold for 1.20 minutes. Mass spectra were acquired by scanning from 100 to 1000. Injection volume was 10 μl. Column temperature was 35° C.

(49) Method 5

(50) In addition to general procedure A: Reversed phase HPLC was carried out on a Sunfire C18 column (3.5 μm, 4.6×100 mm) with an initial flow rate of 0.8 ml/min. Two mobile phases (mobile phase A: 35% 6.5 mM ammonium acetate+30% acetonitrile+35% formic acid (2 ml/1); mobile phase B: 100% acetonitrile) were employed to run a gradient condition from 100% A (hold for 1 minute) to 100% B in 4 minutes, hold at 100% B at a flow rate of 1.2 ml/min for 4 minutes and reequilibrated with initial conditions for 3 minutes. An injection volume of 10 μl was used. Positive ionization mode was used with four different cone voltages (20, 40, 50, 55 V). Mass spectra were acquired by scanning from 100 to 1000 in 0.4 seconds using an interscan delay of 0.1 seconds.

(51) Method 6

(52) In addition to general procedure D: Reversed phase UPLC was carried out on a Waters Acquity BEH (bridged ethylsiloxane/silica hybrid) C18 column (1.7 μm, 2.1×100 mm) with a flow rate of 0.35 ml/min. Two mobile phases (mobile phase A: 95% 7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100% acetonitrile) were employed to run a gradient condition from 90% A and 10% B (hold for 0.5 minutes) to 8% A and 92% B in 3.5 minutes, hold for 2 min and back to the initial conditions in 0.5 min, hold for 1.5 minutes. An injection volume of 2 μl was used. Cone voltages were 20, 30, 45, 60 V for positive ionization mode. Mass spectra were acquired by scanning from 100 to 1000 in 0.2 seconds using an interscan delay of 0.1 seconds.

(53) Method 7

(54) In addition to general procedure D: Reversed phase UPLC was carried out on a Thermo Hypersil Gold C18 column (1.9 μm, 2.1×100 mm) with a flow rate of 0.40 ml/min. Two mobile phases (mobile phase A: 95% 7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100% acetonitrile) were employed to run a gradient condition from 72% A and 28% B (hold for 0.5 minutes) to 8% A and 92% B in 3.5 minutes, hold for 2 min and back to the initial conditions in 0.5 min, hold for 1.5 minutes. An injection volume of 2 μl was used. Cone voltages were 20, 30, 45, 60 V for positive ionization mode. Mass spectra were acquired by scanning from 100 to 1000 in 0.2 seconds using an interscan delay of 0.1 seconds.

(55) Method 8

(56) In addition to general procedure D: Reversed phase UPLC was carried out on a Waters Acquity BEH (bridged ethylsiloxane/silica hybrid) C18 column (1.7 μm, 2.1×100 mm) with a flow rate of 0.35 ml/min. Two mobile phases (mobile phase A: 100% 7 mM ammonium acetate; mobile phase B: 100% acetonitrile) were employed to run a gradient condition from 75% A and 25% B (hold for 0.5 minutes) to 8% A and 92% B in 3.5 minutes, hold for 2 minutes and reequilibrated with initial conditions for 2 minutes. An injection volume of 2 μl was used. Cone voltages were 20, 30, 45, 60 V for positive ionization mode. Mass spectra were acquired by scanning from 100 to 1000 in 0.2 seconds using an interscan delay of 0.1 seconds.

(57) Method 9

(58) In addition to general procedure D: Reversed phase UPLC was carried out on a Thermo Hypersil Gold C18 column (1.9 μm, 2.1×100 mm) with a flow rate of 0.50 ml/min. Two mobile phases (mobile phase A: 95% 7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100% acetonitrile) were employed to run a gradient condition from 40% A and 60% B (hold for 0.5 minutes) to 5% A and 95% B in 3.5 minutes, hold for 2 min and back to the initial conditions in 0.5 min, hold for 1.5 minutes. An injection volume of 2 μl was used. Cone voltages were 20, 30, 45, 60 V for positive ionization mode. Mass spectra were acquired by scanning from 100 to 1000 in 0.2 seconds using an interscan delay of 0.1 seconds.

(59) Method 10

(60) In addition to general procedure A: Reversed phase HPLC was carried out on a Varian Pursuit Diphenyl column (5 μm, 4×100 mm) with a flow rate of 0.8 ml/min. Two mobile phases (mobile phase A: 100% 7 mM ammonium acetate; mobile phase B: 100% acetonitrile) were employed to run a gradient condition from 80% A, 20% B (hold for 0.5 minutes) to 90% B in 4.5 minutes, 90% B for 4 minutes and reequilibrated with initial conditions for 3 minutes. An injection volume of 10 μl was used. Cone voltages were 20, 40, 50, 55 V for positive ionization mode. Mass spectra were acquired by scanning from 100 to 1000 in 0.3 seconds using an interscan delay of 0.05 seconds.

(61) When a compound is a mixture of isomers which give different peaks in the LCMS method, only the retention time of the main component is given in the LCMS table.

D. Pharmacological Examples

MIC90 Determination for Testing Compounds Against M. tuberculosis

(62) Flat-bottom, sterile 96-well plastic microtiter plates were filled with 100 μl of Middlebrook (1×) 7H9 broth medium. Subsequently, an extra 100 μl medium was added to column 2. Stock solutions (200×final test concentration) of compounds were added in 2 μl volumes to a series of duplicate wells in column 2 so as to allow evaluation of their effects on bacterial growth. Serial 2-fold dilutions were made directly in the microtiter plates from column 2 to 11 using a multipipette. Pipette tips were changed after every 3 dilutions to minimize pipetting errors with high hydrophobic compounds. Untreated control samples with (column 1) and without (column 12) inoculum were included in each microtiter plate. Approximately 10000 CFU per well of Mycobacterium tuberculosis (strain H37RV), in a volume of 100 μl in Middlebrook (1×) 7H9 broth medium, was added to the rows A to H, except column 12. The same volume of broth medium without inoculum was added to column 12 in row A to H. The cultures were incubated at 37° C. for 7 days in a humidified atmosphere (incubator with open air valve and continuous ventilation). On day 7 the bacterial growth was checked visually.

(63) The 90% minimal inhibitory concentration (MIC.sub.90) was determined as the concentration with no visual bacterial growth.

(64) Time Kill Assays

(65) Bactericidal or bacteriostatic activity of the compounds can be determined in a time kill assay using the broth dilution method. In a time kill assay on Mycobacterium tuberculosis (strain H37RV), the starting inoculum of M. tuberculosis is 10.sup.6 CFU/ml in Middlebrook (1×) 7H9 broth. The antibacterial compounds are used at the concentration of 0.1 to 10 times the MIC.sub.90. 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.−1 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 is commonly defined as 3-log.sub.10 decrease in number of CFU per ml as compared to untreated inoculum. The potential carryover effect of the drugs is removed by serial dilutions and counting the colonies at highest dilution used for plating.

(66) MIC Values

(67) TABLE-US-00001 MIC90 (μg/ml) LV12076 LV12086 No human 10% human No human 20% human Compound serum serum serum serum Compound 0.5 1 0.25 1 1 0.5 1 0.25 1 Compound 0.06 0.25 0.03 0.25 3 0.06 0.25 0.03 0.25 Isoniazid 0.03 0.03 0.03 0.03 0.03 0.03 0.03 0.03

(68) This experiment was done in microplates; starting from dry powder.

(69) Kill Kinetics

(70) TABLE-US-00002 log CFU/ml (and days) strain compound 0 1 4 7 14 21 H37RV Control 6.35 6.52 6.84 8.00 7.96 9.19 Compound 1 - 6.35 5.57 2.00 2.00 2.00 2.00 0.5 μg/ml Compound 1 - 6.35 6.15 3.08 2.00 2.00 2.00 5 μg/ml Compound 3 - 6.35 5.64 2.60 2.00 2.00 2.00 0.06 μg/ml Compound 3 6.35 5.40 3.32 2.00 2.00 2.00 0.6 μg/ml Isoniazid 1 μg/ml 6.35 3.57 2.00 2.00 2.00 2.00