US20170313721A1

US20170313721A1 - Substituted 1,2,3-triazol-1-yl-methyl-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazoles as anti-mycobacterial agents and a process for the preparation thereof

Info

Publication number: US20170313721A1
Application number: US15/520,799
Authority: US
Inventors: Kushalava Reddy YEMPALLA; Gurunadham MUNAGALA; Samsher SINGH; Sumit Sharma; Inshad Ali Khan; Ram Asrey Vishwakarma; Parvinder Pal Singh
Original assignee: Council of Scientific and Industrial Research CSIR
Current assignee: Council of Scientific and Industrial Research CSIR
Priority date: 2014-10-21
Filing date: 2015-09-09
Publication date: 2017-11-02
Anticipated expiration: 2035-09-16
Also published as: EP3209669A1; US9822126B1; CN107108650B; JP2017531674A; CN107108650A; JP6467042B2; EP3209669B1; WO2016063298A1

Abstract

The present invention relates to new generation of triazole functionality containing 6-nitro-2,3-dihydroimidazooxazole agents for general formula I, their method of preparation and their use as drugs for treatment of tuberculosis, MDR-TB and XDR-TB either alone or in combination with other anti-tubercular agents. In general formula 1, X is selected from a group (CH₂)_nor a direct bond, where n is any number from 1-6, Y is selected from O, S or direct bond, R₁is selected from the group consisting of H, alkyl, substituted alkyl, aryl, substituted aryl, biaryl, substituted biaryl, heterocyclic and substituted heterocyclic, wherein the substituted heterocyclic is selected from any of the following rings consisting of piperazinyl, morpholinyl, piperidyl, pyridyl, triazolyl, triazinyl, pyrimidinyl, pyridazinyl, oxazolyl, furanyl, benzofuranyl, thiophenyl, pyrrolyl, imidazoyl, thiazoyl, quinolinyl, isoquinolinyl, benzooxazolyl and benzothiazolyl and the substitution on aryl and biaryl is selected from the group consisting of F, CI, Br, I, CF₃, OCF₃, OR₁₁, NO₂and alkyl chain from C1 to C14, wherein R₁₁is selected from the group consisting of H, alkyl, phenyl and substituted phenyl.

Description

FIELD OF THE INVENTION

The present invention relates to substituted 1,2,3-triazol-1-yl-methyl-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazolesas anti-mycobacterial agents. The present invention particularly relates to the compounds of 6-nitro-2,3-dihydroimidazooxazole scaffold that have been designed, synthesized and their biological evaluation result for anti-tuberculosis are presented. The present invention relates to novel compounds of general formula I, their method of preparations, and their use as drugs for treatment of tuberculosis.

BACKGROUND OF THE INVENTION

Tuberculosis remains a leading infectious cause of death worldwide and infects about one-third of the world's population. The World Health Organization (WHO) has estimated that if the present conditions remain unchanged, more than 30 million lives will be claimed by TB between 2000 and 2020. In 2012, an estimated 8.6 million people developed TB and 1.3 million died from the disease (including 320 000 deaths among HIV-positive people). TB has also been declared as a global health emergency because of the increase in secondary infections and/or co-infection in cancer and immunocompromised patients (such as those infected with human immunodeficiency virus). The existing lengthy TB therapy and emergence of multidrug resistant TB (MDR-TB) and extensively drug resistant TB (XDR-TB), [BemerMelchior, P.; Bryskier, A.; Drugeon, H. B. J. Antimicrob. Chemother. 2000, 46, 571; Abubaker, J.; Schraufnagel, D. J. Am. Med. Assoc. 2000, 283, 54; Dye. C.; Scheele, S.; Dolin, P.; Pathania, V.; Raviglione, M. C. J. Am. Med. Assoc. 1999, 282, 677] necessitates the development of new and potent anti-tuberculosis agents.
In this direction, we have initiated a medicinal chemistry programme on 6-nitro-2,3-dihydroimidazooxazole scaffold and discovered new potent structures (2954/DEL/2013) and in continuation, in the present invention, new generation triazole functionality containing 6-nitro-2, 3-dihydroimidazooxazole is synthesized and screened for anti-TB activity.

OBJECTIVE OF THE INVENTION

The main objective of the present invention is to provide new generation triazole functionality containing 6-nitro-2, 3-dihydroimidazooxazoles agents for treatment of tuberculosis.
Still another object of the present invention is to provide a process for the preparation of triazole functionality containing 6-nitro-2, 3-dihydroimidazooxazoles.
Still another object of the present invention is to provide treatment against multi-drug resistant (MDR) and extensive drug resistant (XDR) tuberculosis.

SUMMARY OF THE INVENTION

The present invention relates to new generation of triazole functionality containing 6-nitro-2, 3-dihydroimidazooxazole agents, their method of preparation and their use as drugs for treatment of tuberculosis.
Accordingly the present invention provides a compound having a general structure of formula I,
wherein,

- ‘X’ is selected from a group (CH₂)_nor a direct bond, where n is any number from 0, 1, 2 to 6,
- ‘Y’ is selected from a group O, S or direct bond,
- R_Iis selected from the group consisting of H, alkyl, substituted alkyl, aryl, substituted aryl, biaryl, substituted biaryl, heterocyclic and substituted heterocyclic, wherein the substituted heterocyclic is selected from any of the following rings consisting of piperazinyl, morpholinyl, piperidyl, pyridyl, triazolyl, triazinyl, pyrimidinyl, pyridazinyl, oxazolyl, furanyl, benzofuranyl, thiophenyl, pyrrolyl, imidazoyl, thiazoyl, quinolinyl, isoquinolinyl, benzooxazolyl and benzothiazolyl and the substitution on aryl and biaryl is selected from the group consisting of F, Cl, Br, I, CF₃, OCF₃, OR_I1, NO₂and alkyl chain from C1 to C14.
- wherein R_I1is selected from the group consisting of H, alkyl, phenyl, substituted phenyl.
- In an embodiment of the present invention, the representative compound of general formula I comprising:
(R)-2-{[4-(4-trifluoromethoxyphenyl)-1H-1,2,3-triazol-1-yl] methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁, Table 1)
(R)-2-{[4-(4-trifluoromethylphenyl)-1H-1,2,3-triazol-1-yl] methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₂, Table 1)
(R)-2-{[4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₃, Table 1)
(R)-2-{[4-(4-fluoro-3-methylphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₄, Table 1)
(R)-2-{[4-(2,4-difluorophenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₅, Table 1)
(R)-2-{[4-(4-phenoxyphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₆, Table 1)
(R)-2-[(4-pentyl-1H-1,2,3-triazol-1-yl)methyl]-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₇, Table 1)
(R)-2-{[4-(4-trifluoromethoxyphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₈, Table 1)
(R)-2-{[4-(4-trifluoromethylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₉, Table 1)
(R)-2-{[4-(3-chlorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₀, Table 1)
(R)-2-{[4-(4-bromophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₁, Table 1)
(R)-2-{[4-(4-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₂, Table 1)
(R)-2-{[4-(3-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₃, Table 1)
(R)-2-{[4-(2-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₄, Table 1)
(R)-2-{[4-(4-ethylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₅, Table 1)
(R)-2-{[4-(3-fluorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₆, Table 1)
(R)-2-{[4-(2-fluorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₇, Table 1)
(R)-2-{[4-(4-isopropylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₈, Table 1)
(R)-2-methyl-6-nitro-2-((4-((pyridin-2-yloxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₁₉, Table 1)
(R)-2-methyl-6-nitro-2-((4-((p-tolylthio)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₀, Table 1)
(R)-2-methyl-6-nitro-2-((4-(2-(p-tolyloxy)ethyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₁, Table 1)
(R)-2-methyl-2-((4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)methyl)-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₂, Table 1)

In an embodiment of the invention wherein the compound of general formula I, for use in treatment of tuberculosis.
In yet another embodiment of the invention of general formula I, wherein said compound exhibits an in-vitro anti-tuberculosis activity against H₃₇Rv Mycobacterium tuberculosis, MDR-TB (resistant to isoniazid and rifampicin) with MIC values in the range of 0.12 to 32 μg/ml.
In still another embodiment of the invention of general formula I, wherein said compound exhibits an in vitro anti-tuberculosis activity against XDR-TB (resistant to isoniazid, rifampicin, amikacin and moxifloxacin) with MIC values in the range of 0.12 to 32 (μg/ml).
In another embodiment of the invention of general formula I, wherein said compound does not exhibit any cytotoxicity upto 40 μg/ml in macrophage J774 cell line.
The compound of formula I exhibits an in vitro anti-tuberculosis activity against replicating and non-replicating stages of Mycobacterium tuberculosis with MIC values in the range of 0.12 to 32 μg/ml.
The compound of formula I exhibits an in vitro anti-tuberculosis activity against XDR Mycobacterium tuberculosis (resistant to isoniazid, rifampicin, amikacin and moxifloxacin), MDR-TB (resistant to isoniazid and rifampicin) with MIC values in the range of 0.12 to 32 μg/ml and the compound does not exhibit any cytotoxicity upto 40 μg/ml in macrophage J774 cell line.
The present invention also provides a process for preparation of the compound of formula I wherein the said process comprising the steps:

- i) reacting a compound of formula 8 in an organic solvent selected from group consisting of toluene, acetonotrile, DMF, dichloroethane or any combination thereof in the presence of an azide source selected from sodiumazide, trimethylsilylazide and tetrabutyl ammonium bromide at a temperature in the range of 25° C. to 80° C. for a period ranging between 1 h to 3 h to obtain the desired compound of formula 9.

- ii) reacting the compound of formula 9 with a base selected from a group consisting of sodium hydride, cesium carbonate, potassium carbonate or any combination thereof in an organic solvent selected from a group consisting of toluene, acetonotrile, DMF, tetrahydrofuran or any combination thereof in the presence of at a temperature in the range of 10° C.-25° C. for a period of 1 h to 12 h to obtain a desired compound of formula 10.

- iii) reacting the compound of formula 10 with a compound of formula selected from the group consisting of formula 13(a-k) or 14(a-g) or 15(a-d) in 1:1 tert-BuOH/H₂O mixture in the presence of sodiumascorbate and CuSO₄at a temperature in the range of 10° C. to 25° C. for a period of 1 h to 12 h to obtain the desired compound of formula I.

List of Abbreviations:

ATCC: american type culture collection
AcOH: acetic acid
CFU: colony forming units
DMAP: 4-dimethylaminopyridine
DCM: dichloromethane
d: doublet
dd: doublet of doublet
Et: ethyl
ESI: electron spray ionisation
FCS: fetal calf serum
H₃₇Rv: a well characterised virulent strain of Mycobacterium tuberculosis
h: hours
IC₅₀: half maximal inhibitory concentration
J: coupling constant
MIC: minimum inhibitory concentration
MS: mass spectrometry
ml: millilitre
MHz: mega hertz
m: multiplet
MDR-TB: Multi drug resistant tuberculosis
Me: methyl
min: minutes
m/z: mass to charge ratio
MTB: Mycobacterium tuberculosis
NMP: N-methylpyrrolidinone
Rif^R: rifampicin resistant tuberculosis
RPMI: rosewell park memorial institute medium
R_f: retention factor
s: singlet
TFA: trifluoroacetic acid
TLC: thin layer chromatography
TB: Tuberculosis
TDR-TB: Total drug resistant tuberculosis
t: triplet
tert: tertiary
WHO: world health organization
XDR-TB: Extensive drug resistant tuberculosis
μg: microgram
¹H NMR: proton nuclear magnetic resonance

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to new generation of triazole functionality containing 6-nitro-2, 3-dihydroimidazooxazole agents, their method of preparation and their use as drugs for treatment of tuberculosis.
The present invention describes a compound having general structure of formula I
wherein

- ‘X’ is selected from a group (CH₂)_nor a direct bond, where n is any number from 0, 1, 2 to 6,
- ‘Y’ is selected from a group O, S or direct bond,
- RI is selected from the group consisting of H, alkyl, substituted alkyl, aryl, substituted aryl, biaryl, substituted biaryl, heterocyclic and substituted heterocyclic, wherein the substituted heterocyclic is selected from any of the following rings consisting of piperazinyl, morpholinyl, piperidyl, pyridyl, triazolyl, triazinyl, pyrimidinyl, pyridazinyl, oxazolyl, furanyl, benzofuranyl, thiophenyl, pyrrolyl, imidazoyl, thiazoyl, quinolinyl, isoquinolinyl, benzooxazolyl and benzothiazolyl and the substitution on aryl and biaryl is selected from the group consisting of F, Cl, Br, I, CF3, OCF3, ORI1, NO2 and alkyl chain from C1 to C14.
- wherein R_I1is selected from the group consisting of H, alkyl, phenyl, substituted phenyl.

The most preferred compounds of general formula I are:

(R)-2-{[4-(4-trifluoromethoxyphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁, Table 1)
(R)-2-{[4-(4-trifluoromethylphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₂, Table 1)
(R)-2-{[4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₃, Table 1)
(R)-2-{[4-(4-fluoro-3-methylphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₄, Table 1)
(R)-2-{[4-(2,4-difluorophenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₅, Table 1)
(R)-2-{[4-(4-phenoxyphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₆, Table 1)
(R)-2-[(4-pentyl-1H-1,2,3-triazol-1-yl)methyl]-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₇, Table 1)
(R)-2-{[4-(4-trifluoromethoxyphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₈, Table 1)
(R)-2-{[4-(4-trifluoromethylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₉, Table 1)
(R)-2-{[4-(3-chlorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₀, Table 1)
(R)-2-{[4-(4-bromophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I_ii, Table 1)
(R)-2-{[4-(4-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₂, Table 1)
(R)-2-{[4-(3-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₃, Table 1)
(R)-2-{[4-(2-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₄, Table 1)
(R)-2-{[4-(4-ethylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₅, Table 1)
(R)-2-{[4-(3-fluorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₆, Table 1)
(R)-2-{[4-(2-fluorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₇, Table 1)
(R)-2-{[4-(4-isopropylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₈, Table 1)
(R)-2-methyl-6-nitro-2-((4-((pyridin-2-yloxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₁₉, Table 1)
(R)-2-methyl-6-nitro-2-((4-((p-tolylthio)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₀, Table 1)
(R)-2-methyl-6-nitro-2-((4-(2-(p-tolyloxy)ethyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₁, Table 1)
(R)-2-methyl-2-((4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)methyl)-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₂, Table 1)

The compounds of general formula I, are useful in treatment of tuberculosis. The compound of general formula I, exhibits an in vitro anti-tuberculosis activity against H₃₇Rv Mycobacterium tuberculosis, MDR-TB (resistant to isoniazid and rifampicin) with MIC values in the range of 0.12 to 32 μg/ml.
The compound of general formula I, exhibits an in vitro anti-tuberculosis activity against XDR-TB (resistant to isoniazid, rifampicin, amikacin and moxifloxacin) with MIC values in the range of 0.12 to 32 (μg/ml).
The compound of general formula I does not exhibit any cytotoxicity upto 40 μg/ml in macrophage J774 cell line.
An embodiment of the invention; provides a process for the preparation of a compound of formula 9, wherein the process step comprising of the reaction of compound of formula 8 in an organic solvent selected from toluene, acetonotrile, DMF, dichloroethaneor any combination thereof in the presence of azide source selected from sodiumazide, trimethylsilylazide, tetrabutyl ammonium bromide or any combination thereof at a temperature in the range of 25° C. to 80° C. for a period of 1 h to 3 h to obtain the desired compound of formula 9.
In another embodiment of the invention a process for the preparation of the compound of formula 10, wherein the process step comprising of the reaction of compound of formula 9 in an organic solvent selected from toluene, acetonotrile, DMF, tetrahydrofuran or any combination thereof in the presence of base selected from sodium hydride, cesium carbonate, potassium carbonate or any combination thereof at a temperature in the range of 10° C. to 25° C. for a period of 1 h to 12 h to obtain the desired compound of formula 10.
In another embodiment of the invention, a process for the preparation of the compound of general formula I, wherein the process step comprising the reacting compound of formula 10 with a compound of formula selected from the group consisting of formula 13 or 14 or 15 in 1:1 tert-BuOH/H₂O mixture in the presence of sodium ascorbate and CuSO₄at a temperature in the range of 10° C. to 25° C. for a period of 1 h to 12 h to obtain the desired compound of general formula I.
The present invention discloses process for synthesis of the fragments A and B which in turn can be used for synthesis of compounds of general formula I. The entire synthesis of compound I is illustrated by Reaction Schemes 1 to 4 as given below: —

EXAMPLES

Synthesis of Compounds:

The following examples are given by way of illustrating the present invention and should not be construed to limit the scope of the invention:

R)-3-(2-Chloro-4-nitro-1H-imidazol-1-yl)-2-hydroxy-2-methylpropylmethanesulfonate (8)

The synthesis of compound 8 was successfully synthesized from a starting material 4-Nitroimidiazole 1 as shown in scheme 1 by following known procedure (Sasaki, H.; Haraguchi, Y.; Itotani, M.; Kuroda, H.; Hashizume, H.; Tomishige, T.; Kawasaki, M.; Matsumoto, M.; Komatsu, M.; Tsubouchi, H. J. Med. Chem. 2006, 49, 7854.

Example 1

(R)-1-Azido-3-(2-chloro-4-nitro-1H-imidazol-1-yl)-2-methylpropan-2-ol (9)

To a solution of (R)-3-(2-Chloro-4-nitro-1H-imidazol-1-yl)-2-hydroxy-2-methylpropyl methanesulfonate (8) (10 mmol) in DMF (20 mL) was added sodium azide (30 mmol) and tetrabutyl ammonium bromide (1 mmol). After the solution was stirred at 80° C. for 3 h, the reaction mixture was extracted with ethyl acetate twice, and the combined organic layer was washed with brine, dried over sodium sulphate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography in 5% DCM/Ethyl acetate to give the compound 9 as yellow colour solid. The compound 9 is also prepared using the process as described above using different solvents such as toluene, acetonitrile or dichloroethane at 60-80° C. for a period of 2-4 hr as given in the following table.


Reactant	Solvent	Temp ° C.	Time	Reagents	Product

(R)-3-(2-Chloro-4-	Toluene	60	2	Sodium azide	(R)-1-Azido-3-(2-
nitro-1H-imidazol-1-					chloro-4-nitro-1H-
yl)-2-hydroxy-2-					imidazol-1-y1)-2-
methylpropyl					methylpropan-2-ol (9)
methanesulfonate (8)
(R)-3-(2-Chloro-4-	Acetonitrile	60	4	Trimethylsily	(R)-1-Azido-3-(2-
nitro-1H-imidazol-1-				lazide	chloro-4-nitro-1H-
y1)-2-hydroxy -2-					imidazol-1-y1)-2-
methylpropyl					methylpropan-2-ol (9)
methanesulfonate (8)
(R)-3-(2-Chloro-4-	dichloroethane	80	2	Sodium azide	(R)-1-Azido-3-(2-
nitro-1H-imidazol-1-					chloro-4-nitro-1H-
yl)-2-hydroxy-2-					imidazol-1-y1)-2-
methylpropyl					methylpropan-2-ol (9)
methanesulfonate (8)

Example 2

(R)-2-(Azidomethyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (10)

To a solution of (R)-1-Azido-3-(2-chloro-4-nitro-1H-imidazol-1-yl)-2-methylpropan-2-ol 9)(10 mmol) in DMF (20 mL) was added cesium carbonate (30 mmol) at below 15° C. portion wise, After the solution was stirred for 12 h at 25° C., The reaction mixture was poured into the ice water and extracted with ethyl acetate twice, and the combined organic layer was washed with brine, dried over sodium sulphate, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give the compound 10 as yellow colour solid.
The reaction is also carried out by using the different solvent such as toluene, acetonitrile or tetrahydrofurane at 25° C. for a period of 6-12 hr. to produce the compound 10.


Reactant	Solvent	Tem. ° C.	Time	Reagents	Product

(R)-1-Azido-3-(2-	DMF	25	6	Sodium	(R)-2-(Azidomethyl)-
chloro-4-nitro-1H-				hydride	2-methy1-6-nitro-2,3-
imidazol-1-y1)-2-					dihydroimidazo [2,1-
methylpropan-2-ol (9)					b] oxazole (10)
(R)-1-Azido-3-(2-	Toluene	25	6	Sodium	(R)-2-(Azidomethyl)-
chloro-4-nitro-1H-				hydride	2-methy1-6-nitro-2,3-
imidazol-1-y1)-2-					dihydroimidazo [2,1-
methylpropan-2-ol (9)					b] oxazole (10)
(R)-1-Azido-3-(2-	Acetonitrile	25	12	Potassium	(R)-2-(Azidomethyl)-
chloro-4-nitro-1H-				carbonate	2-methy1-6-nitro-2,3-
imidazol-1-y1)-2-					dihydroimidazo [2,1-
methylpropan-2-ol (9)					b] oxazole (10)
(R)-1-Azido-3-(2-	tetrahydro-	25	12	Sodium	(R)-2-(Azidomethyl)-
chloro-4-nitro-1H-	furan			carbonate	2-methy1-6-nitro-2,3-
imidazol-1-y1)-2-					dihydroimidazo [2,1-
methylpropan-2-ol (9)					b] oxazole (10)

Example 3

General Procedure for the Preparation of Compounds (I₁to I₂₂):
(R)-2-(Azidomethyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (10) (1 mmol) and 13 (a-k) or 14 (a-g) or 15 (a-d) suspended in 6 mL of a 1:1 tert-BuOH/H₂O mixture. While the mixture was being stirred, sodium ascorbate (0.1 mmol) was added, followed by CuSO₄pentahydrate (0.02 mmol). Left stirring for 12 h at 25° C., after which time it was diluted with water, and the solid was filtered off. The crude was purified by silica gel column chromatography to give the compounds I₁to I₂₂.

(R)-2-Methyl-6-nitro-2-((4-(4-(trifluoromethoxy)phenyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₁, Table 1, Scheme 3)

TLC (EtOAc:DCM 1:9): R_f=0.3; ¹H NMR (400 MHz, Acetone) δ 8.50 (s, 2H), 8.01 (t, J=4.5 Hz, 7H), 7.39 (d, J=8.0 Hz, 5H), 5.10 (q, J=14.9 Hz, 6H), 4.66 (d, J=11.2 Hz, 3H), 4.42 (d, J=11.2 Hz, 3H), 1.79 (s, 7H); MS (ESI+): m\z 410.0950.

(R)-2-Methyl-6-nitro-2-((4-(4-(trifluoromethyl)phenyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₂, Table 1, Scheme 3)

TLC (EtOAc:DCM 1:9): R_f=0.45; ¹H NMR (400 MHz, DMSO) δ 8.74 (s, 1H), 8.07 (t, J=4.0 Hz, 3H), 7.81 (d, J=8.3 Hz, 2H), 5.05 (d, J=14.8 Hz, 1H), 4.98 (d, J=14.8 Hz, 1H), 4.44 (d, J=11.3 Hz, 1H), 4.26 (d, J=11.3 Hz, 1H), 1.63 (s, 3H); MS (ESI+): m\z 394.1001.

(R)-2-((4-(4-Fluorophenyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6nitro-2,3-dihydro imidazo[2,1-b]oxazole (I₃, Table 1, Scheme 3)

TLC (EtOAc:DCM 1:9): R_f=0.35; ¹H NMR (400 MHz, DMSO) δ 8.54 (s, 1H), 8.05 (s, 1H), 7.87 (dd, J=8.7, 5.5 Hz, 2H), 7.28 (t, J=8.9 Hz, 2H), 5.01 (d, J=14.8 Hz, 1H), 4.95 (d, J=14.8 Hz, 1H), 4.42 (d, J=11.3 Hz, 1H), 4.24 (d, J=11.2 Hz, 1H), 1.63 (s, 3H); MS (ESI+): m\z 344.1033.

(R)-2-((4-(4-Fluoro-3-methylphenyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₄, Table 1, Scheme 3)

TLC (EtOAc:DCM 1:9): R_f=0.5; ¹H NMR (400 MHz, DMSO) δ 8.47 (s, 1H), 8.02 (s, 1H), 7.72 (d, J=7.0 Hz, 1H), 7.69-7.60 (m, 1H), 7.19 (t, J=9.1 Hz, 1H), 5.00 (d, J=14.8 Hz, 1H), 4.93 (d, J=14.8 Hz, 1H), 4.41 (d, J=11.3 Hz, 1H), 4.23 (d, J=11.3 Hz, 1H), 2.27 (s, 3H), 1.63 (s, 3H); MS (ESI+): m\z 358.1190.

(R)-2-((4-(2,4-Difluorophenyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₅, Table 1, Scheme 3)

TLC (EtOAc:DCM 1:9): R_f=0.25; ¹H NMR (500 MHz, Acetone) δ 8.35 (d, J=3.6 Hz, 2H), 8.21 (dd, J=15.5, 8.8 Hz, 2H), 7.81 (s, 2H), 7.20-7.11 (m, 4H), 5.15 (q, J=14.9 Hz, 6H), 4.68 (d, J=11.1 Hz, 3H), 4.43 (d, J=11.1 Hz, 3H), 1.79 (s, 8H); MS (ESI+): m\z 362.0939.

(R)-2-Methyl-6-nitro-2-((4-(4-phenoxyphenyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₆, Table 1, Scheme 3)

TLC (EtOAc:DCM 1:9): R_f=0.25; ¹H NMR (400 MHz, DMSO) δ 8.47 (s, 1H), 8.02 (s, 1H), 7.82 (d, J=8.7 Hz, 2H), 7.41 (t, J=7.9 Hz, 2H), 7.16 (t, J=7.4 Hz, 1H), 7.05 (dd, J=8.1, 6.0 Hz, 4H), 5.00 (d, J=14.8 Hz, 1H), 4.93 (d, J=14.8 Hz, 1H), 4.42 (d, J=11.3 Hz, 1H), 4.24 (d, J=11.3 Hz, 1H), 1.63 (s, 3H); MS (ESI+): m\z 418.1390.

(R)-2-[(4-Pentyl-1H-1,2,3-triazol-1-yl)methyl]-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (I₇, Table 1, Scheme 3)

TLC (EtOAc:DCM 1:9): R_f=0.5; ¹H NMR (400 MHz, Acetone) δ 8.18 (s, 1H), 7.80 (s, 1H), 4.62 (d, J=11.2 Hz, 2H), 4.40 (d, J=11.2 Hz, 2H), 2.25 (t, 2H), 1.75 (s, 3H), 1.21 (m, 4H), 1.15 (m, 2H), 0.91 (m, 3H); MS (ESI+): m\z 320.1579.

(R)-2-Methyl-6-nitro-2-((4-((4-(trifluoromethoxy)phenoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₈, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.45; ¹H NMR (400 MHz, Acetone) δ 8.18 (s, 1H), 7.79 (s, 1H), 7.28 (d, J=9.2 Hz, 2H), 7.14 (d, J=9.2 Hz, 2H), 5.22 (s, 2H), 5.08 (q, J=14.9 Hz, 2H), 4.65 (d, J=11.2 Hz, 1H), 4.42 (d, J=11.2 Hz, 1H), 1.78 (s, 3H); MS (ESI+): m\z 440.1056.

(R)-2-Methyl-6-nitro-2-((4-((4-(trifluoromethyl)phenoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₉, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.15; ¹H NMR (400 MHz, DMSO) δ 8.23 (s, 1H), 8.05 (s, 1H), 7.65 (d, J=8.7 Hz, 2H), 7.20 (d, J=8.5 Hz, 2H), 5.23 (s, 2H), 4.97 (q, J=14.8 Hz, 2H), 4.39 (d, J=11.3 Hz, 1H), 4.23 (d, J=11.3 Hz, 1H), 1.59 (s, 3H); MS (ESI+): m\z 424.1107.

(R)-2-((4(3-Chlorophenoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₁₀, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.25; ¹H NMR (400 MHz, DMSO) δ 8.21 (s, 1H), 8.05 (s, 1H), 7.30 (t, J=8.2 Hz, 1H), 7.11 (t, J=2.0 Hz, 1H), 6.99 (ddd, J=10.4, 8.2, 1.6 Hz, 2H), 5.14 (s, 2H), 4.96 (q, J=14.8 Hz, 2H), 4.39 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.3 Hz, 1H), 1.59 (s, 3H); MS (ESI+): m\z 390.0843.

(R)-2-(4-((4-Bromophenoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₁₁, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.35; ¹H NMR (400 MHz, DMSO) δ 8.20 (s, 1H), 8.04 (s, 1H), 7.44 (d, J=8.6 Hz, 2H), 6.98 (d, J=8.4 Hz, 2H), 5.11 (s, 2H), 4.96 (q, J=14.8 Hz, 2H), 4.38 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.2 Hz, 1H), 1.58 (s, 3H); MS (ESI+): m\z 434.0338.

(R)-2-Methyl-6-nitro-2-((4-((p-tolyloxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₁₂, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.3; ¹H NMR (400 MHz, DMSO) δ 8.17 (s, 1H), 8.05 (s, 1H), 7.08 (d, J=8.2 Hz, 2H), 6.88 (d, J=8.4 Hz, 2H), 5.07 (s, 2H), 5.02-4.88 (m, 2H), 4.38 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.3 Hz, 1H), 2.23 (s, 3H), 1.59 (s, 3H); MS (ESI+): m\z 370.1390.

(R)-2-Methyl-6-nitro-2-((4-((m-tolyloxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₁₃, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.3; ¹H NMR (400 MHz, DMSO) δ 8.17 (s, 1H), 8.03 (s, 1H), 7.15 (t, J=7.8 Hz, 1H), 6.77 (dd, J=16.5, 7.7 Hz, 3H), 5.07 (s, 2H), 4.95 (q, J=14.8 Hz, 2H), 4.38 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.3 Hz, 1H), 2.26 (s, 3H), 1.58 (s, 3H); MS (ESI+): m\z 370.1390.

(R)-2-Methyl-6-nitro-2-((4-((o-tolyloxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₁₄, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.35; ¹H NMR (400 MHz, Acetone) δ 8.14 (s, 7H), 7.78 (s, 6H), 7.12 (dt, J=22.2, 7.9 Hz, 26H), 6.85 (t, J=6.9 Hz, 8H), 5.17 (s, 21H), 5.07 (q, J=14.9 Hz, 27H), 4.62 (d, J=11.2 Hz, 12H), 4.40 (d, J=11.2 Hz, 12H), 2.13 (s, 20H), 1.77 (s, 36H); MS (ESI+): m\z 370.1390.

(R)-2-((4-((4-Ethylphenoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₁₅, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.5; ¹H NMR (400 MHz, Acetone) δ 8.11 (s, 1H), 7.79 (s, 1H), 7.13 (d, J=8.4 Hz, 2H), 6.92 (d, J=8.5 Hz, 2H), 5.19-4.98 (m, 6H), 4.63 (d, J=11.2 Hz, 2H), 4.40 (d, J=11.1 Hz, 2H), 2.58 (dd, J=15.1, 7.5 Hz, 2H), 1.76 (s, 3H), 1.18 (t, J=7.6 Hz, 14H); MS (ESI+): m\z 384.1546.

(R)-2-((4-((3-Fluorophenoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₁₆, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.2; ¹H NMR (400 MHz, DMSO) δ 8.21 (s, 1H), 8.03 (s, 1H), 7.30 (dd, J=15.3, 7.7 Hz, 1H), 6.93-6.72 (m, 3H), 5.12 (s, 2H), 4.96 (q, J=14.7 Hz, 2H), 4.38 (d, J=11.1 Hz, 1H), 4.22 (d, J=11.2 Hz, 1H), 1.59 (s, 3H); MS (ESI+): m\z 374.1139.

(R)-2-((4-((2-Fluorophenoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₁₇, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.3; ¹H NMR (400 MHz, Acetone) δ 8.18 (s, 1H), 7.80 (s, 1H), 7.29 (td, J=8.6, 1.6 Hz, 1H), 7.17-7.09 (m, 2H), 6.96 (ddd, J=7.8, 7.0, 1.5 Hz, 1H), 5.26 (s, 3H), 5.07 (q, J=14.9 Hz, 4H), 4.62 (d, J=11.2 Hz, 2H), 4.40 (d, J=11.2 Hz, 2H), 1.75 (s, 5H); MS (ESI+): m\z 374.1139.

(R)-2-((4-((4-Isopropylphenoxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₁₈, Table 1, Scheme 4)

TLC (EtOAc:DCM 1:9): R_f=0.35; ¹H NMR (400 MHz, DMSO) δ 8.18 (s, 1H), 8.07 (s, 1H), 7.14 (d, J=8.5 Hz, 2H), 6.91 (d, J=8.6 Hz, 2H), 5.07 (s, 2H), 4.96 (q, J=14.8 Hz, 2H), 4.39 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.2 Hz, 1H), 2.83 (dt, J=13.8, 6.8 Hz, 1H), 1.58 (s, 3H), 1.17 (d, J=6.9 Hz, 6H); MS (ESI+): m\z 398.1703.

(R)-2-Methyl-6-nitro-2-((4-((pyridin-2-yloxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₁₉, Table 1, Scheme 5)

TLC (EtOAc:DCM 1:9): R_f=0.4; ¹H NMR (400 MHz, DMSO) δ 8.19 (s, 1H), 8.04 (s, 1H), 7.98 (m, 1H), 7.25-7.38 (m, 3H), 5.10 (s, 2H), 4.95 (q, J=14.8 Hz, 2H), 4.38 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.3 Hz, 1H), 1.58 (s, 3H); MS (ESI+): m\z 357.1186.

(R)-2-Methyl-6-nitro-2-((4-((p-tolylthio)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₂₀, Table 1, Scheme 5)

TLC (EtOAc:DCM 1:9): R_f=0.3; ¹H NMR (400 MHz, DMSO) δ 8.17 (s, 1H), 8.05 (s, 1H), 7.08 (d, J=8.2 Hz, 2H), 6.88 (d, J=8.4 Hz, 2H), 5.07 (s, 2H), 5.02-4.88 (m, 2H), 4.38 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.3 Hz, 1H), 2.23 (s, 3H), 1.59 (s, 3H); MS (ESI-F): m\z 386.1161.

(R)-2-Methyl-6-nitro-2-((4-(2-(p-tolyloxy)ethyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (I₂₁, Table 1, Scheme 5)

TLC (EtOAc:DCM 1:9): R_f=0.35; ¹H NMR (400 MHz, DMSO) δ 8.17 (s, 1H), 8.05 (s, 1H), 7.08 (d, J=8.2 Hz, 2H), 6.88 (d, J=8.4 Hz, 2H), 5.02-4.88 (m, 2H), 4.38 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.3 Hz, 1H), 4.12 (d, J=4.8 Hz, 2H), 2.32 (d, J=4.8 Hz, 2H), 2.23 (s, 3H), 1.59 (s, 3H); MS (ESI+): m\z 384.1546.

(R)-2-Methyl-2-((4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)methyl)-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (I₂₂, Table 1, Scheme 5)

TLC (EtOAc:DCM 1:9): R_f=0.25; ¹H NMR (400 MHz, DMSO) δ 8.16 (s, 1H), 8.06 (s, 1H), 5.04 (s, 2H), 5.02-4.88 (m, 2H), 4.38 (d, J=11.3 Hz, 1H), 4.22 (d, J=11.3 Hz, 1H), 4.09-4.18 (d, 4H), 3.36-3.48 (m, 4H), 1.58 (s, 3H); MS (ESI+): m\z 349.1499.

Biological Evaluation

Example 1

In Vitro Activity of Compounds I_Ito I₂₂Against M. tuberculosis H₃₇Rv and Two Clinical Isolates (M. tuberculosis MDR & M. tuberculosis XDR)
MIC Determination:
MIC was determined by broth dilution method against M. tuberculosis H₃₇Rv (ATCC 27294; American Type Culture Collection, Manassas, Va.), M. tuberculosis MDR (resistant to isoniazid and rifampicin) and M. tuberculosis XDR (resistant to isoniazid, rifampicin, amikacin and moxifloxacin). The bacterial strains were grown for 10 to 15 days in Middlebrook 7H9 broth (Difco Laboratories, Detroit, Mich.) supplemented with 0.5% (v/v) glycerol, 0.25% (v/v) Tween 80 (Himedia, Mumbai India), and 10% ADC (albumin dextrose catalase, Becton Dickinson, Sparks, Md.) under shaking conditions at 37° C. in 5% CO₂to facilitate exponential-phase growth of the organism. Bacterial suspension was prepared by suspending M. tuberculosis growth in normal saline containing 0.5% tween 80 and turbidity was adjusted to 1 McFarland standard which is equivalent to 1.5×10⁷CFU/ml. The 2-fold serial dilutions of compounds I₁to I₂₂were prepared in Middle brook 7H9 (Difco laboratories) for M. tuberculosis in 100 μl per well in 96-well U bottom microtitre plates (Tarson, Mumbai, India). The above-mentioned bacterial suspension was further diluted in the growth media and 100 μl volume of this diluted inoculum was added to each well of the plate resulting in the final inoculum of 5×10⁵CFU/ml in the well and the final concentrations of compound I_Ito I₂₂ranged from 0.015 to 32 μg/ml (0.015, 0.03, 0.06, 0.12, 0.25, 0.5, 1, 2, 4, 8, 16, 32). The plates were incubated at 37° C. for 3-weeks in 5% CO₂. The plates were read visually and the minimum concentration of the compound showing no turbidity was recorded as MIC.

Results:

i) The compounds of general formula I (compounds I₁to I₂₂), were screened against both replicating & non-replicating stages of M. tuberculosis, wherein 7 compounds I₁₀, I₁₁, I₁₃, I₁₄, I₁₇, I₁₈and I₂₁showed MIC value of <1.0 μg/ml (results provided in Table 2). Three compounds I₁₁, I₁₇and I₂₁showed very potent MIC of 0.12, 0.25 and 0.25 μg/ml against replicating stages of M. Tuberculosis and MIC of 0.25, 0.5 and 0.25 against non-replicating stages of M. Tuberculosis. The results are given in Table 2.
ii) The compounds of general formula I (compounds I₁to I₂₂), were screened against both multi-drug and extensive-drug resistant strains of M. tuberculosis, wherein five compounds I₁₁, I₁₇, I₁₉, I₂₀and I₂₁showed MIC value of <1.0 μg/ml. Three compounds I₁₁, I₁₇and I₂₁showed very potent MIC of 0.12, 0.5 and 0.25 μg/ml against multi and extensive-drug resistant strains of M. Tuberculosis. The results are given in Table 2.

Example 2

Cytotoxicity Assay of Compounds I₁to I₂₂:

Cell Culture:

The study was carried out using macrophage J774 cells line (ATCC-USA). Cells were grown in Rosewell Park Memorial Institute Medium (RPMI) containing 10% fetal calf serum (FCS) and supplemented with 75 mg/litre penicillin, 100 mg/litre streptomycin, 110 mg/litre Sodium pyruvate, 2.38 gm/litre HEPES, 0.05 mM 2 β-mercaptoethanol, and 2 gm/litre NaHCO₃, in a humidified atmosphere in 5% CO₂at 37° C., and were sub-cultured at 1:4 ratio once a week.

Cell Treatment:

Cells were plated at a density of 3×10⁴cells/cm²and maintained in culture medium for 12 hours. Cells were seeded onto 96-well flat bottom plates and FCS was reduced to 5% for the experiment. Stock solutions of compounds 9 to 37 were prepared fresh to avoid oxidation. Cells were incubated with the compounds (40 μg/ml) for 24 hrs.

Cytotoxicity Assays:

After the completion of incubation, the medium was removed and cell viability was evaluated by assaying for the ability of functional mitochondria to catalyze the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to form formazan salt by mitochondrial dehydrogenases, and determined by Elisa reader at 450 nm (Multiskan Spectrum; Thermo Electron Corporation, USA). Percentage cytotoxicity was calculated with respect to the untreated cells.

Results:

Compounds I₁to I₂₂were not toxic up to 40 μg/ml concentration and the cytotoxicity assay results are shown in Table 2.

TABLE 1

Structure of representative compounds I₁to I₂₂of general formula I

Entries	Codes	Structures

1	I₁

2	I₂

3	I₃

4	I₄

5	I₅

6	I₆

7	I₇

8	I₈

9	I₉

10	I₁₀

11	I₁₁

12	I₁₂

13	I₁₃

14	I₁₄

15	I₁₅

16	I₁₆

17	I₁₇

18	I₁₈

19	I₁₉

20	I₂₀

21	I₂₁

22	I₂₂

TABLE 2

Anti-tuberculosis activity and cytotoxicity of representative
compounds of general formula I (I₁to I₂₂)

MIC (μg/ml)

	Compound	M. tb	Non-replicating	MIC			Cytotoxicity
S. No.	code	H₃₇Rv	strain	(Rif^R)	MDR	XDR	(μg/ml)

1.	I₁	2	4	2	2	1	>40
2.	I₂	2	8	4	2	2	>40
3.	I₃	1	2	2	2	2	>40
4.	I₄	2	4	1	4	2	>40
5.	I₅	1	8	4	1	4	>40
6.	I₆	2	2	1	1	1	>40
7.	I₇	2	4	8	2	8	>40
8.	I₈	2	4	2	2	2	>40
9.	I₉	1	2	4	4	4	>40
10.	I₁₀	0.5	1	2	2	4	>40
11.	I₁₁	0.12	0.25	0.12	0.12	0.25	>40
12.	I₁₂	1	2	8	2	8	>40
13.	I₁₃	0.5	0.5	1	1	2	>40
14.	I₁₄	0.5	2	1	1	1	>40
15.	I₁₅	2	2	4	2	4	>40
16.	I₁₆	1	2	2	2	2	>40
17.	I₁₇	0.25	0.5	1	0.5	1	>40
18.	I₁₈	0.5	1	2	1	4	>40
19.	I₁₉	1	2	0.5	0.25	0.5	>40
20.	I₂₀	2	1	0.25	0.5	2	>40
21.	I₂₁	0.25	0.25	1	0.25	0.25	>40
22.	I₂₂	2	1	2	2	2	>40

Claims

1. A compound of general formula I,

wherein

‘X’ is selected from a group (CH₂)_nor a direct bond, where n is any number from 0, 1, 2 to 6,

‘Y’ is selected from a group O, S or direct bond,

R_Iis selected from the group consisting of H, alkyl, substituted alkyl, aryl, substituted aryl, biaryl, substituted biaryl, heterocyclic and substituted heterocyclic, wherein the substituted heterocyclic is selected from any of the following rings consisting of piperazinyl, morpholinyl, piperidyl, pyridyl, triazolyl, triazinyl, pyrimidinyl, pyridazinyl, oxazolyl, furanyl, benzofuranyl, thiophenyl, pyrrolyl, imidazoyl, thiazoyl, quinolinyl, isoquinolinyl, benzooxazolyl and benzothiazolyl and the substitution on aryl and biaryl is selected from the group consisting of F, Cl, Br, I, CF₃, OCF₃, OR_I1, NO₂and alkyl chain from C1 to C14,

wherein R_I1is selected from the group consisting of H, alkyl, phenyl, substituted phenyl.

2. The compound of formula I as claimed in claim 1, wherein the compound is selected from the group consisting of the following compounds:

(R)-2-{[4-(4-trifluoromethoxyphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁)

(R)-2-{[4-(4-trifluoromethylphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₂)

(R)-2-{[4-(4-fluorophenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₃)

(R)-2-{[4-(4-fluoro-3-methylphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₄)

(R)-2-{[4-(2,4-difluorophenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₅)

(R)-2-{[4-(4-phenoxyphenyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₆)

(R)-2-[(4-pentyl-1H-1,2,3-triazol-1-yl)methyl]-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₇)

(R)-2-{[4-(4-trifluoromethoxyphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₈)

(R)-2-{[4-(4-trifluoromethylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₉)

(R)-2-{[4-(3-chlorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₀)

(R)-2-{[4-(4-bromophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₁)

(R)-2-{[4-(4-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₂)

(R)-2-{[4-(3-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₃)

(R)-2-{[4-(2-methylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₄)

(R)-2-{[4-(4-ethylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₅)

(R)-2-{[4-(3-fluorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₆)

(R)-2-{[4-(2-fluorophenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₇)

(R)-2-{[4-(4-isopropylphenoxy)methyl)-1H-1,2,3-triazol-1-yl]methyl}-2,3-dihydro-2-methyl-6-nitroimidazo[2,1-b]oxazole (compound I₁₈)

(R)-2-methyl-6-nitro-2-((4-((pyridin-2-yloxy)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₁₉)

(R)-2-methyl-6-nitro-2-((4-((p-tolylthio)methyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₀)

(R)-2-methyl-6-nitro-2-((4-(2-(p-tolyloxy)ethyl)-1H-1,2,3-triazol-1-yl)methyl)-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₁)

(R)-2-methyl-2-((4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)methyl)-6-nitro-2,3-dihydroimidazo[2,1-b]oxazole (compound I₂₂)

3. The compound of general formula I as claimed in claim 1, for use in the treatment of tuberculosis.

4. The compound of formula I as claimed in claim 1, wherein said compound exhibits an in vitro anti-tuberculosis activity against replicating and non-replicating stages of Mycobacterium tuberculosis with MIC values in the range of 0.12 to 32 μg/ml.

5. The compound of formula I as claimed in claim 1, wherein said compound exhibits an in vitro anti-tuberculosis activity against XDR Mycobacterium tuberculosis (resistant to isoniazid, rifampicin, amikacin and moxifloxacin), MDR-TB (resistant to isoniazid and rifampicin) with MIC values in the range of 0.12 to 32 μg/ml and the compound does not exhibit any cytotoxicity up to 40 μg/ml in macrophage J774 cell line.

6. A process for preparation of the compound of formula I as claimed in claim 1 wherein the said process comprising the steps:

i) reacting a compound of formula 8 in an organic solvent and in the presence of an azide source at a temperature in the range of 25° C. to 80° C. for a period ranging between 1 h to 3 h to obtain a compound of formula 9;

ii) reacting the compound of formula 9 in an organic solvent and in the presence of a base at a temperature in the range of 10° C.-25° C. for a period of 1 h to 12 h to obtain a compound of formula 10;

iii) reacting the compound of formula 10 with a compound of formula selected from the group consisting of formula 13 (a-k) or 14 (a-g) or 15 (a-d) in 1:1 tert-BuOH/H₂O mixture in the presence of sodiumascorbate and CuSO₄at a temperature in the range of 10° C. to 25° C. for a period of 1 h to 12 h to obtain the compound of formula I.

7. The process as claimed in step i) of claim 6, wherein the organic solvent is selected from toluene, acetonitrile, DMF, dichloroethane or any combination thereof.

8. The process as claimed in step i) of claim 6, wherein the azide source is selected from sodiumazide, trimethylsilylazide, tetrabutyl ammonium bromide or any combination thereof.

9. The process as claimed in step ii) of claim 6, wherein the base is selected from sodium hydride, cesium carbonate, potassium carbonate or any combination thereof.

10. The process as claimed in step ii) of claim 6, wherein the organic solvent is selected from toluene, acetonitrile, DMF, tetrahydrofuran or any combination thereof.

11. The compound of formula I as claimed in claim 2, wherein said compound exhibits an in vitro anti-tuberculosis activity against replicating and non-replicating stages of Mycobacterium tuberculosis with MIC values in the range of 0.12 to 32 μg/ml.

12. The compound of formula I as claimed in claim 2, wherein said compound exhibits an in vitro anti-tuberculosis activity against XDR Mycobacterium tuberculosis (resistant to isoniazid, rifampicin, amikacin and moxifloxacin), MDR-TB (resistant to isoniazid and rifampicin) with MIC values in the range of 0.12 to 32 μg/ml and the compound does not exhibit any cytotoxicity up to 40 μg/ml in macrophage J774 cell line.