-By

Aalpesh Kachhadiya
2009H108403H
What is Tuberculosis?
Infectious disease caused by the Mycobacterium
tubercule species
Primarily target lungs, but also kidney lung apices,
bone marrow, kidneys, and meninges
About 32% world’s population [2 billion people] are
infected with TB.
In India alone, one person dies of TB every minute.
Types of tuberculosis:
 MDR tuberculosis
 XDR tuberculosis
 Latent tuberculosis
Stages of M.tuberculosis Infection
How Mtb tackle immune system?
CURRENT TB THERAPY
The treatment duration is a minimum of 6 months,
with 4 drugs (isoniazid, rifampin, pyrazinamide, and
ethambutol) typically given daily for the first 2
months and with 2 drugs (isoniazid and rifampin)
administered for 4 additional months.
Compliance-the emergence of drug resistance.

Potential for drug-drug interactions, primarily those

between rifampin and many of the anti-HIV drugs.
Current therapy:
ISONIAZID

AN T
T
RIFAMPIN

E S I S
R
PYRAZINAMIDE

ETHAMBUTOL

STREPTOMYCIN
Objectives for TB Drug Development:
Simplify treatment/reduce treatment duration

Have an acceptable toxicity profile

Be an active against MDR/XDR TB

Be useful in HIV –infected patients with TB

Be active against latent TB

No Drug –drug interaction

 Global TB Drug Portfolio
September 2005
Discovery Preclinical Clinical Testing

Carboxylates Nitrofuranylamides Diamine SQ-109 Diarylquinoline TMC207

TB Alliance, Wellesley College NIAID, University of Tennessee Sequella Inc. Johnson & Johnson

Nitroimidazole Analogs Dipiperidines (SQ-609) Gatifloxacin

Cell Wall Inhibitors
NIAID, Novartis Institute for Tropical Diseases, OFLOTUB Consortium, Lupin, NIAID TBRU,
Colorado State University, NIAID Sequella Inc.
TB Alliance Tuberculosis Research Centre, WHO TDR

Dihydrolipoamide Acyltransferase Non-Fluorinated Quinolone Moxifloxacin

Novel Antibiotic Class Bayer Pharmaceuticals, CDC TBTC, Johns
Inhibitors TaiGen
GlaxoSmithKline, TB Alliance Hopkins University, NIAID TBRU, TB Alliance
Cornell University, NIAID

InhA Inhibitors Picolinamide Imidazoles Synthase Inhibitor FAS20013 Nitroimidazole PA-824

GlaxoSmithKline, TB Alliance NIAID, TAACF) FASgen Inc. Chiron Corporation, TB Alliance

Translocase I Inhibitors Nitroimidazo-oxazole OPC-67683

Isocitrate Lyase Inhibitors (ICL) Pleuromutilins Sequella Inc., Sankyo Otsuka
GlaxoSmithKline, TB Alliance GlaxoSmithKline, TB Alliance

Macrolides Quinolones Nitroimidazo-oxazole Back-up Pyrrole LL-3858

TB Alliance, University of Illinois at Chicago KRICT/ Yonsei University, NIAID, Otsukai Lupin Limited
TAACF, TB Alliance

Methyltransferase Inhibitors Screening and Target Identification

Anacor Pharmaceuticals AstraZeneca

Natural Products Exploration

BIOTEC, California State University, ITR, NIAID, Thiolactomycin Analogs
TAACF, University of Auckland NIAID, NIH
M. Spigelman, GATB
Newer therapy

The American Journal of Medicine (2008) 121, 835-844

MOA of New Drugs in clinical trial:
 Dormant Mtb target

Active Mtb target

Targets for/of New Anti-TB drug

L,D-transpeptidase
Two transpeptidase present:
D,D transpeptidase-43linkage generation 
Inhibited by β lactam antibiotics
L,D transpeptidase – 33 linkage generation  80% of
pepetide linkage
Inhibition of both enzymes ,attenuated growth,virulence
and ability to persist
Also sensitized Mtb to amoxicillin-clavulanate –invitro
and invivo
GTPases:
 PI3K play essential role in
maturation of phagosomes
 LAM: prevents PI3P generation
on phagosomes
 SapM: act as PI3P phosphatase
to remove this lipid from
phagosomes
 Rab22a-member of family
Rabs5(GTPase)-prevent
phagosome maturation
 FtsZ-another assembling
GTPase enzyme in Mtb
MEP(2 C -methyl- D-erythritol 4-phosphate)
Isoprenoid is essential which either functions as toxins,growth
inhibitors or secondary metabolites
Mtb isoprenoids:
 Pol-P “linkerunit” between two essential cell wall

components,arabinogalactan and peptidoglycan

 Menaquinone:essential for electron transport chain

All isoprenoids are derived from the repitive condensation of

Isopentyl diphosphate and dimethylallyl
diphosphate:synthesised only through MEP pathway
Known inhibitors for MEP enzymes:
Summary of targets:
Remodeling of Existing antibacterial class
References:
 Zhenkun Ma et.al- ‘Global tuberculosis drug development pipeline: the need and
the reality’, Series: Tuberculosis 5
 Gyanu Lamichhane, ‘Review-Novel targets in M. tuberculosis : search for new drugs’,
Cel Press
 Anil Koul et.al- ‘The challenge of new drug discovery for tuberculosis,
Nature:Review’, doi:10.1038/nature09657
 Spigelman, ‘New Tuberculosis Therapeutics: A Growing Pipeline’, J Infect Dis.-2007
,S28-34
 Laxman S. Meena, Review:”Guanosine triphosphatases as novel therapeutic targets in
tuberculosis”, International Journal of Infectious Diseases 14 (2010) e682–e687
 Hyungjin Eoh, Review:”The Mycobacterium tuberculosis MEP (2 C -methyl- D-erythritol
4-phosphate) pathway as a new drug target”, Tuberculosis 89 (2009) 1–11
 Jossy van den Boogaard, Minireview: ” New Drugs against Tuberculosis: Problems,
Progress, and Evaluation of Agents in Clinical Development”, ANTIMICROBIAL
AGENTS AND CHEMOTHERAPY, Mar. 2009, p. 849–862
  Working group on New TB Drug: http://www.newtbdrugs.org/project.php?id=25, date-
24th April 2011
Change of target:GSK and TB alliance
The glyoxylate pathway uses isocitrate lyase (ICL) and
malate synthase to incorporate carbon during growth
of microorganisms on acetate or fatty acids as the
primary carbon source.
ICL:project was discontinued due to lack of
druggability of the ICL targets.
Structural analysis indicated that malate synthase is a
much more druggable target by virtue of its deeper
and more hydrophobic binding domain.
Mycolic acid cyclopropanation:
Mycolic acid :Major constitute of cell wall

Cyclopropanation require to unite different mycolic acid

molecule
 Multiple iso-enzymes present.

Dioctylamine : broad inhibition of cyclopropanation-

bacteriostatic at 6μM
DprE1/DprE2:
DprE1 heterodimerizes with DprE2 to form active enzyme
Converts : to incorporate into the cellwall
decaprenylphosphoryl ribose decaprenylphosphoryl
arabinose
Inhibitor: BTZ043 (1-4ng/ml)- benzothiazinone class
: Dinitrobenzamides
Showed similar potency in various strain of TB:
 Drug sensitive
 XDR

 MDT

 extensively resistant strain

Mycothiol ligase:
Mycothiol, low molecular weight thiol that protects Mtb from
toxicity of antibiotics and oxidative stresses

Dequalinium chloride
High affinity
Inhibition of growth :0.3-1.2μg/mL
ATP phosphoribosyl transferase:
Histidine biosynthesis pathway enzymes
Histidine essential for the growth of Mtb
Conversion of:

5 –phospho-α-D-ribose 1- HisG L- Histidine

diphosphate

Inhibited by:
Nitrobenzothiazole Phosporibosyl -ATP
ATP synthase:
Involve in the synthesis of ATP

Diaryl quinoloine compound(R201910):Affinity for the α & β

subunit of the enzyme.

Inhibit the enzyme in dose dependent manner

Active in dormant and drug resistant strain also

20,000 times less affinity for Human ATP synthase

Currently in Clinical trials designated asTMC-207

Protein processing: deformylase and methionine
aminopeptidase
mRNA translation begins with the incorporation of formylated

methionine at the amino (N) terminus.

This residue is removed in a two-step process:

o deformylation by peptide deformylase

 Inhibitor: LBK-611 displayed a MIC in the micromolar range

o hydrolysis of methionine by methionine aminopeptidase

 Inhibitor: 2,3-dichloro-1,4 napthoquinones

 Human beings are devoid of above enzyme, hence good target for anti-TB activity
Isocitrate lyase:
Role: fatty acid synthesis, virulence characteristics
Inhibitor: 3-nitro-propionate
Inhibition of isocitrate lyase activity causes genetic
loss of icl1 and icl2 gene.
Results into loss in ability to:
 Grow in fatty acids,
 Loss of virulence
 Growth defect during acute and chronic phase of
infection

Recent news:
The Proteasome complex:
Role: degradation of oxidized proteins,
Proteasome activity required to maintain infection
during chronic phase
Morphology:
 Accessory ATPase enzyme associated with
proteoasome Mpa.
 Presence of catalytically active site threonine residue
 Inhibitor:
 Threonine site : Oxathiazol-2-one
Synergizes with the immune control of Mtb infection
DosR(DevR)
After engulfment by macrophage, Mtb experience
hypoxia and stress condition(NO)
Hence increased levels of DosR(DevR) which are
transcription factor, that binds to major groove of
DNA as a tetramer – activates dormancy regulon
physiological adaption
Presence of 48 genes
Inhibition of DosR alone might be effective in
controlling growth or have synergistic effects with
other anti-TB drug
Maltosyltransferase GlgE
Accumalation of Maltose-1-phosphate is toxic to Mtb
Adversely affect the Electron transport chain .
GlgE utilizes maltose-1-phosphate to elongate 1,4-
glucan chains.
Transcriptional profile was similar to that elicited by
Pot.cyanide
Gut flora and mammals lack proteins homologous to
GlgE ,hence suitable drug target