anti-TB Agents Research Articles

Design, synthesis and structure-activity relationship study of wollamide B; a new potential anti TB agent.

Wollamide B is a cationic antimycobacterial cyclohexapeptide that exhibits activity against Mycobacterium bovis (M. bovis) (IC50 of 3.1 μM). Aiming to define its structural activity relationship (SAR), optimizing potency and pharmacokinetic properties, libraries of analogues were synthesized following a standard Fmoc-based solid phase peptide synthesis approach. The antimycobacterial activities of wollamide B and all the synthesized analogues were tested against Mycobacterium tuberculosis (Mtb) H37Rv. Parallely, in vitro drug metabolism and pharmacokinetic (ADME) profiling was done for the synthesized compounds to evaluate their drug likeness. Among the 25 synthesized wollamides five of them showed potent activities with MICs ≤ 3.1 μM and found to be nontoxic against human HepG2 cells up to 100 μM. The results of the in vitro ADME profiling revealed the remarkable plasma stability and very good aqueous solubility of the class in general while the metabolic stability was found to be moderate to low. Of particular note, compounds 7c (MIC = 1.1 μM) and 13c (0.6 μM) that exhibited good balance of antimycobacterial activity vs. optimal pharmacokinetic properties could be used as a new lead for further development.

Revisiting the mutant prevention concentration to guide dosing in childhood tuberculosis.

The mutant prevention concentration (MPC) is a well-known concept in the chemotherapy of many bacterial infections, but is seldom considered in relation to tuberculosis (TB) treatment, as the required concentrations are generally viewed as unachievable without undue toxicity. Early studies revealed single mutations conferring high MICs of first- and second-line anti-TB agents; however, the growing application of genomics and quantitative drug susceptibility testing in TB suggests a wide range of MICs often determined by specific mutations and strain type. In paediatric TB, pharmacokinetic studies indicate that despite increasing dose recommendations, a proportion of children still do not achieve adult-derived targets. When considering the next stage in anti-TB drug dosing and the introduction of novel therapies for children, we suggest consideration of MPC and its incorporation into pharmacokinetic studies to more accurately determine appropriate concentration targets in children, to restrict the growth of resistant mutants and better manage drug-resistant TB.

Adolescent tuberculosis: A challenge and opportunity to prevent community transmission

Abstract Introduction Adolescents may present with adult-type pulmonary tuberculosis (TB), including cavity disease in upper lobes and smear-positive sputum, which involves a significant transmission risk for social and family contacts. Patients and methods A retrospective (2007–2012) observational study of a case series of TB was conducted in children and adolescents ( 12 years at diagnosis were compared. Results The series consisted of 124 patients (56.5% males, median age: 4.0 years). In half of the cases, the patient was of immigrant origin and TB was diagnosed after clinical–radiological suspicion, intra-thoracic disease being the most common (91.9%). Cultures yielded positive results in one third of cases (37.9%) and isolates were sensitive to oral first-line anti-TB agents in 100%. Median (interquartile range) duration of treatment was 6 (6–9) months, directly observed therapy was needed in 10 patients, and there was a satisfactory outcome after treatment in 98.4%. Among adolescents, TB was more prevalent in females (63.2%) and immigrant patients (68.4%), comorbidity at diagnosis and lung cavity forms were more common, and the source case was identified only in 21.1% of the patients. Conclusion Adult-type pulmonary TB is common among adolescents, may be associated with underlying medical conditions, and is often diagnosed late, posing a significant transmission risk to the community.

Validation of Mycobacterium tuberculosis dihydroneopterin aldolase as a molecular target for anti-tuberculosis drug development

An early step of target validation in antimicrobial drug discovery is to prove that a gene coding for a putative target is essential for pathogen's viability. However, little attention has been paid to demonstrate the causal links between gene essentiality and a particular protein function that will be the focus of a drug discovery effort. This should be considered an important step in target validation since a growing number of proteins are found to exhibit multiple and unrelated tasks. Here, we show that the Mycobacterium tuberculosis (Mtb) folB gene is essential and that this essentiality depends on the dihydroneopterin aldolase/epimerase activities of its protein product, the FolB protein from the folate biosynthesis pathway. The wild-type (WT) MtFolB and point mutants K99A and Y54F were cloned, expressed, purified and monitored for the aldolase, epimerase and oxygenase activities using HPLC. In contrast to the WT MtFolB, both mutants have neither aldolase nor epimerase activities in the conditions assayed. We then performed gene knockout experiments and showed that folB gene is essential for Mtb survival under the conditions tested. Moreover, only the WT folB sequence could be used as a rescue copy in gene complementation studies. When the sequences of mutants K99A or Y54F were used for complementation, no viable colonies were obtained, indicating that aldolase and/or epimerase activities are crucial for Mtb survival. These results provide a solid basis for further work aiming to develop new anti-TB agents acting as inhibitors of the aldolase/epimerase activities of MtFolB.

An ethnobotanical survey of plants used traditionally to treat tuberculosis in the eastern region of O.R. Tambo district, South Africa

Worldwide, tuberculosis (TB) is a major human killer claiming about 1.5 million lives per annum. Medicinal plants are believed to serve as a source for products that can serve as anti-TB agents. Despite the importance of plants for human health and subsistence in both developed and developing countries, loss of biodiversity-based cultural knowledge and traditions is a commonly reported phenomenon, therefore, documenting such knowledge before it disappears completely is a necessity. The study was carried out to document medicinal plants that are traditionally used for the treatment of TB in the eastern region of O.R. Tambo district, Eastern Cape Province of South Africa. Information about plants (names, parts and methods of preparation) that are used for treating TB was gathered from traditional medical practitioners (TMP) using questionnaires. Twenty-four plant species belonging to 19 families that are used for the treatment of TB were revealed by this study. The Apiaceae, Amaryllidaceae, Asparagaceae, Compositae, and Leguminosae were the most prominent, represented by two species each. Out of the 24 plants reported, Protorhus longifolia (Bernh.) Engl (Anacardiaceae), Phymaspermum acerosum Källersjö (Compositae) and Strychnos henningsii Gilg (Loganiaceae) were the most frequently mentioned species, and were reported for the first time for TB treatment. Most of the plants (91%) documented are administered orally; the root (54%) is the most common plant part used, while decoctions and infusions are the main preparatory methods. Indigenous knowledge of medicinal plants used in the treatment of TB exists in the eastern region of O.R. Tambo district, and TMP still play an important role in delivering primary health care services. The ethnobotanical information about the plant species mentioned in this study may serve as baseline data for future studies on their pharmacological effects and to identify those that have potential in the development of anti-TB drugs.

Bedaquiline and delamanid: Sharing the indian experience

Currently, India is home to the highest number of multi-drug resistant tuberculosis (MDR-TB) patients in the world. Until now, MDR-TB was being treated with repurposed drugs which are toxic and are required to be given for up to 2 years. Two new oral anti-TB drugs, bedaquiline and delamanid have received approval in 2012 and 2014 respectively. While presenting the pharmacology, evidence of efficacy, WHO recommendations, current status of access as well as future directions regarding these two novel anti-TB agents, this paper delves into the India narrative recounting the short clinical experience thus far.

Recent Trends in Treatment of Multidrug Resistant-Tuberculosis-A Review

Drug resistance is reduction in effectiveness of an anti-microbial agents used in inhibiting the microorganisms or curing a disease. Mycobacterium tuberculosis is responsible for causing tuberculosis can acquire multiple drug resistance by not responding to Isoniazid and Rifampicin the two most powerful anti-TB agents. The complications of drug resistance in TB elevates due to some of the risk factors like inadequate treatment compliance, noncompliance of the patients to the treatment. Proper diagnosis of the disease and switching to modified drug therapy may improve the treatment outcome. The drug treatment involves usage of combination of drugs Isoniazid, Rifampicin, Ethambutol along with other antimicrobial agents, and nutritional therapy, immunotherapy. In this review we discussed about the occurrence, treatment of multi drug resistance tuberculosis along with drugs under trials and nutritional requirements.

Tuberculosis and Transplantation.

Mycobacterium tuberculosis is a major opportunistic pathogen in transplant recipients. Compared to that in the general population, the frequency of tuberculosis (TB) is 10 to 40 times higher in hematopoietic stem cell transplant (HSCT) recipients and 20 to 74 times higher in solid-organ transplant (SOT) recipients. Transplant recipients with TB are also more likely to develop disseminated disease, have longer time to definitive diagnosis, require more invasive diagnostic procedures, and experience greater anti-TB treatment-related toxicity than the general population. Specific risk factors for TB in SOT recipients include previous exposure to M. tuberculosis (positive tuberculin skin tests and/or residual TB lesions in pretransplant chest X ray) and the intensity of immunosuppression (use of antilymphocyte antibodies, type of basal immunosuppression, and intensification of immunosuppressive therapy for allograft rejection). Risk factors in HSCT recipients are allogeneic transplantation from an unrelated donor; chronic graft-versus-host disease treated with corticosteroids; unrelated or mismatched allograft; pretransplant conditioning using total body irradiation, busulfan, or cyclophosphamide; and type and stage of primary hematological disorder. Transplant recipients with evidence of prior exposure to M. tuberculosis should receive treatment appropriate for latent TB infection. Optimal management of active TB disease is particularly challenging due to significant drug interactions between the anti-TB agents and the immunosuppressive therapy. In this chapter, we address the epidemiology, clinical presentation, diagnostic considerations, and management strategies for TB in SOT and HSCT recipients.

Spectinamides are effective partner agents for the treatment of tuberculosis in multiple mouse infection models.

Objectives: New drug regimens employing combinations of existing and experimental antimicrobial agents are needed to shorten treatment of tuberculosis (TB) in humans. The spectinamides are narrow-spectrum semisynthetic analogues of spectinomycin, modified to avoid intrinsic efflux by Mycobacterium tuberculosis. Spectinamides, including lead 1599, have been previously shown to exhibit a promising therapeutic profile in mice as single agents. Here we explore the in vivo activity of lead spectinamides when combined with other agents. Methods: The efficacy of 1599 or 1810 was tested in combination in three increasingly advanced TB mouse models. Mice were infected by aerosol and allowed to establish acute or chronic infection, followed by treatment (≤4 weeks) with the spectinamides alone or in two- and three-drug combination regimens with existing and novel therapeutic agents. Bacteria were enumerated from lungs by plating for cfu. Results: Herein we show the following: (i) 1599 exhibits additive or synergistic activity with most of the first-line agents; (ii) 1599 in combination with rifampicin and pyrazinamide or with bedaquiline and pyrazinamide promotes significantly improved efficacy in the high-dose aerosol model; (iii) 1599 enhances efficacy of rifampicin or pyrazinamide in chronically infected BALB/c mice; and (iv) 1599 is synergistic when administered in combination with rifampicin and pyrazinamide in the C3HeB/FeJ mouse model showing caseous necrotic pulmonary lesions. Conclusions: Spectinamides were effective partner agents for multiple anti-TB agents including bedaquiline, rifampicin and pyrazinamide. None of these in vivo synergistic interactions was predicted from in vitro MIC chequerboard assays. These data support further development of the spectinamides as combination partners with existing and experimental anti-TB agents.

Mycobacterial cell wall biosynthesis: a multifaceted antibiotic target

Mycobacterium tuberculosis (Mtb), the etiological agent of tuberculosis (TB), is recognized as a global health emergency as promoted by the World Health Organization. Over 1 million deaths per year, along with the emergence of multi- and extensively-drug resistant strains of Mtb, have triggered intensive research into the pathogenicity and biochemistry of this microorganism, guiding the development of anti-TB chemotherapeutic agents. The essential mycobacterial cell wall, sharing some common features with all bacteria, represents an apparent 'Achilles heel' that has been targeted by TB chemotherapy since the advent of TB treatment. This complex structure composed of three distinct layers, peptidoglycan, arabinogalactan and mycolic acids, is vital in supporting cell growth, virulence and providing a barrier to antibiotics. The fundamental nature of cell wall synthesis and assembly has rendered the mycobacterial cell wall as the most widely exploited target of anti-TB drugs. This review provides an overview of the biosynthesis of the prominent cell wall components, highlighting the inhibitory mechanisms of existing clinical drugs and illustrating the potential of other unexploited enzymes as future drug targets.

Eco-Friendly and Highly Efficient Synthesis, Including Multigram Synthesis, of Aldehyde Isonicotinoyl Hydrazones Using Sonochemistry

Background: The isoniazid has been a key compound in first-line tuberculosis (TB) treatment, usually in combination with other drugs. Following on from the use of isoniazid itself, various derivatives of isoniazid have also been investigated as anti-TB agents and other diseases. This work aims to synthesize, multigram scale, salicylaldehyde isonicotinoyl hydrazone from isoniazid and salicylaldehyde, using ultrasound. Additionally, we have been interested in establishing a general, smaller-scale, ultrasonic driven synthesis of arylaldehyde isoniazid N-acylhydrazones, particularly for use in our biological studies. Methods: A Multiwave Eco-Sonics QR750 ultrasonic generator (20 KHz, 750 W) equipped with a converter/ transducer, and titanium oscillator (horn, diameter = 4 mm) was used for the ultrasonic irradiantion. Compounds were characterized by melting points, infrared spectra, mass spectra (CG/MS), electron microscopy, X-ray powder diffraction, solid-state 13C and solution NMR spectra. Results: This work describes a green chemistry protocol, multigram scale (ca 4 mole) and highly efficient synthesis utilizing ultrasonic irradiation of salicylaldehyde isonicotinoyl hydrazone, a widely used compound in biological studies. With a short reaction time of 30 min, and a facile work up, involving washing with water, the important bioactive product was obtained in a high yield, 98.7%, and in a high purity 99.43%. Also prepared using ultrasonic irradiation with short reaction times, in high yields and high purities, but on smaller scales, were a series arylaldehyde isoniazid N-acylhydrazones, all of which exhibit promising activity against cancer and tuberculosis. Comparisons with the classical procedures indicate advantages in the ultrasonic irradiated methodology. Conclusion: We report the high yielding ultrasonic synthesis of pure salicylaldehyde isonicotinoyl hydrazone on a 4 mole scale. In addition, a series of ten isoniazid N-acylhydrazone derivatives has been obtained using ultrasonic irradiation. The ultrasonic procedures are simple, safe, with short reaction times, and produce high yields. Keywords: Green chemistry, isoniazid, N-acylhydrazone, SIH, bioactive, sonochemistry.

Modelling the Transitional Dynamics of Mycobacterium Tuberculosis Strain

The World Health Organization’s targets of eliminating Tuberculosis (TB) by 2050 is challenged by the emergence and spread of drug resistance TB. However, the traditional mechanism of resistance is that of acquired resistance, whereby the mycobacterium Tuberculosis (MTB) strain develops mutations under selective pressure of insufficient drug therapy. These mutations have the tendency of changing the drug target protein, restricting the bacteria to the anti-TB agent. We propose a discrete state markov chain model with three disease states: Drug Susceptible (DS), Multi Drug Resistant (MDR) and Extra Drug Resistant (XDR) to further study the transitional dynamics of the MTB strain. The study made use of a retrospective data on resistant pattern to first line and second line anti TB drugs. The structural properties of the model established life expectancies of DS and MDR strains as well as the probability of first resistance of the DS strain. Key estimates were assessed by the bootstrapping procedure which converged in estimates to the actual data. If the experiment were repeated infinitely many times, in 95 out of 100, the interval 2.782 x 10-7 to 0.018 will contain the true probability of first mutation of the DS strain. A key contribution of this study is the revealing therapeutic cycle of the treatment regime of the TB disease based on the TB progression data which saw the period after the 20th cycle of the treatment being prominent in some key strain dynamics. These findings may also help explain further the pharmacodynamic properties of the "first line" anti-Tuberculosis drugs for enhance TB treatment. Journal of Medical and Biomedical Sciences (2016) 5(2), 13-23

Naturally Occurring Anti-TB Agents: Isolation, Chemical Transformations and In Vitro Antitubercular Activities of Secondary Metabolites of Rhizomes of Alpinia galanga

A bioactivity-guided chemical examination of the acetone extract of the rhizomes of Alpinia galanga led to the isolation of six secondary metabolites, eucalyptol derivative (1) and phenylpropanoids (2–6). The structures of all of the isolated compounds (1–6) were elucidated on the basis of their spectral data. The isolated compounds (1–6) were in vitro assayed against active and dormant phenotypes of Mycobacterium tuberculosis H37Ra, respectively. Interestingly, 1′S-1′-acetoxychavicol acetate (2) showed good antitubercular activities against both active and dormant phenotypes of M. tuberculosis with IC50 values of 1.04 µM and 2.69 µM, respectively. Tsuji-Trost and homodimerization reactions of the active compound (2) respectively resulted in the formation of two analogues, 7 and 8. Both of these synthesized analogues were also found to be active in vitro against active [IC50 s of 3.24 and 3.87 µM, respectively, for compounds 7 and 8] and dormant [IC50 s of 8.33 and 2.41 µM, respectively, for compounds 7 and 8] phenotypes of M. tuberculosis H37Ra, respectively.

Interactions of linezolid and second-line anti-tuberculosis agents against multidrug-resistant Mycobacterium tuberculosis in vitro and in vivo

The objectives of this study were to evaluate the interactions between linezolid (LZD) and second-line anti-tuberculosis (TB) agents in susceptible and multidrug-resistant (MDR) TB in vitro, and to validate the in vitro results in a murine TB model. The minimum inhibitory concentrations of LZD and seven second-line anti-TB drugs against H37Rv and three multidrug-resistant clinical isolates were determined by Alamar Blue assay, and the interaction patterns of LZD and the seven second-line anti-TB agents against the four isolates were studied using a dynamic checkerboard method. The activities of these combinations against Mycobacterium tuberculosis were evaluated in a murine model of TB. The combination of LZD + capreomycin exhibited partial synergism for three of four isolates, LZD + para-aminosalicylic acid exhibited partial synergism for two of four isolates, and LZD + levofloxacin and LZD + amikacin exhibited partial synergism for one of four isolates; all other combinations showed indifference or an additive effect in vitro. The activities of six combinations and the standard regimen rifampicin + isoniazid + pyrazinamide were investigated in a murine model of TB (infection with H37Rv). Significant reductions in colony-forming units (CFU) were found in LZD + capreomycin and LZD + clofazimine groups when the CFU in the lungs on day 0 (the day of beginning treatment) was compared with the CFU in the lungs after 2 months of treatment. These combinations of LZD and second-line anti-TB drugs were all active against MDR-TB with indifference or an additive effect, except LZD + capreomycin, which showed partial synergy.

Bis-biguanide dihydrochloride inhibits intracellular replication of M. tuberculosis and controls infection in mice.

While there is an urgent need to develop new and effective drugs for treatment of tuberculosis (TB) and multi-drug resistant TB (MDR-TB), repurposing FDA (U.S. Food and Drug Administration) -approved drugs for development of anti-TB agents may decrease time and effort from bench to bedside. Here, we employed host cell-based high throughput screening (HTS) assay to screen and characterize FDA-approved, off-patent library drugs for anti-Mycobacterium tuberculosis (MTB) activities. The cell-based HTS allowed us to identify an anti-cancer drug of bis-biguanide dihydrochloride (BBD) as potent anti-mycobacteria agent. Further characterization showed that BBD could inhibit intracellular and extracellular growth of M. smegmatis and slow-growing M. bovis BCG. BBD also potently inhibited replication of clinically-isolated MTB and MDR-TB strains. The proof-of-concept study showed that BBD treatment of MTB-infected mice could significantly decrease CFU counts in the lung and spleen. Notably, comparative evaluation showed that MTB CFU counts in BBD-treated mice were lower than those in rifampicin-treated mice. No apparent BBD side effects were found in BBD-treated mice. Thus, our findings support further studies to develop BBD as a new and effective drug against TB and MDR-TB.

Bioluminescent Reporters for Rapid Mechanism of Action Assessment in Tuberculosis Drug Discovery.

The tuberculosis (TB) drug discovery pipeline is fueled by compounds identified in whole-cell screens against the causative agent, Mycobacterium tuberculosis Phenotypic screening enables the selection of molecules that inhibit essential cellular functions in live, intact bacilli grown under a chosen in vitro condition. However, deducing the mechanism of action (MOA), which is important to avoid promiscuous targets, often requires significant biological resources in a lengthy process that risks decoupling medicinal chemistry and biology efforts. Therefore, there is a need to develop methods enabling rapid MOA assessment of putative "actives" for triage decisions. Here, we describe a modified version of a bioluminescence reporter assay that allows nondestructive detection of compounds targeting either of two macromolecular processes in M. tuberculosis: cell wall biosynthesis or maintenance of DNA integrity. Coupling the luxCDABE operon from Photorhabdus luminescens to mycobacterial promoters driving expression of the iniBAC operon (PiniB-LUX) or the DNA damage-inducible genes, recA (PrecA-LUX) or radA (PradA-LUX), provided quantitative detection in real time of compounds triggering expression of any of these promoters over an extended 10- to 12-day incubation. Testing against known anti-TB agents confirmed the specificity of each reporter in registering the MOA of the applied antibiotic in M. tuberculosis, independent of bactericidal or bacteriostatic activity. Moreover, profiles obtained for experimental compounds indicated the potential to infer complex MOAs in which multiple cellular processes are disrupted. These results demonstrate the utility of the reporters for early triage of compounds based on the provisional MOA and suggest their application to investigate polypharmacology in known and experimental anti-TB agents.

Genetics-directed drug discovery for combating Mycobacterium tuberculosis infection

Mycobacterium tuberculosis (Mtb), the pathogen of tuberculosis (TB), is one of the most infectious bacteria in the world. The traditional strategy to combat TB involves targeting the pathogen directly; however, the rapid evolution of drug resistance lessens the efficiency of this anti-TB method. Therefore, in recent years, some researchers have turned to an alternative anti-TB strategy, which hinders Mtb infection through targeting host genes. In this work, using a theoretical genetic analysis, we identified 170 Mtb infection-associated genes from human genetic variations related to Mtb infection. Then, the agents targeting these genes were identified to have high potential as anti-TB drugs. In particular, the agents that can target multiple Mtb infection-associated genes are more druggable than the single-target counterparts. These potential anti-TB agents were further screened by gene expression data derived from connectivity map. As a result, some agents were revealed to have high interest for experimental evaluation. This study not only has important implications for anti-TB drug discovery, but also provides inspirations for streamlining the pipeline of modern drug discovery.

Development of a prediction system for anti-tuberculosis drug-induced liver injury in Japanese patients.

Drug-induced liver injury (DILI) is a common adverse drug reaction in patients receiving antituberculosis (anti-TB) treatment. Among the anti-TB agents, isoniazid (INH) is the primary drug that causes hepatotoxicity in TB patients with DILI. Previous reports in several populations have consistently demonstrated an association between polymorphisms in the N-acetyltransferase 2 (NAT2) gene, which is responsible for INH hepatic metabolism, and a risk of DILI in TB patients. In this study, the genetic and baseline clinical data from 366 Japanese patients with TB (73 patients with DILI and 293 without DILI) were used to develop a system to predict DILI risk due to anti-TB agents. The distribution of the NAT2 acetylator status among the TB patients with DILI was 31 (42.5%), 29 (39.7%), and 13 (17.8%) for rapid, intermediate, and slow acetylators, respectively. A significant association was observed between NAT2 slow acetylators and DILI risk (odds ratio 4.32, 95% confidence interval 1.93–9.66, P value=5.56×10−4). A logistic regression model based on age and NAT2 genotype revealed that the area under the curve for the receiver-operating characteristic curve was 0.717. The findings demonstrated that slow NAT2 acetylator status is a significant predictor of the risk of DILI by anti-TB agents, and a personalized anti-TB treatment approach may aid in making treatment decisions and reducing the incidence of DILI.

First-line treatment of paediatric tuberculosis requires consideration of disease type and severity followed by careful monitoring

The first-line treatment of paediatric tuberculosis (TB) involves long-term treatment with regimens containing multiple anti-TB agents (i.e. isoniazid, rifampicin, pyrazinamide and/or ethambutol). The choice of treatment regimen is based on the clinical manifestation of TB, disease severity and the risk of antibacterial resistance. Paediatric patients should be carefully monitored for treatment compliance, response to treatment and drug-related adverse events, of which hepatotoxicity is the most common.

Hepatoprotective properties of oleanolic and ursolic acids in antitubercular drug-induced liver damage.

To estimate to what extent the mixture of ursolic acid and oleanolic acid, in addition to the antitubercular standard regime, affects the hepatotoxicity profile. Liver injury was induced in male BALB/c mice by administering, per os and daily for 11 weeks, a combination of anti-Tubercular (anti-TB) agents Rifampicin (10mg/kg), Isoniazid (10mg/kg), and Pyrazinamide (30mg/kg). The ursolic acid and oleanolic acid mixture at doses of 100 or 200μg/mouse/day was subcutaneously injected throughout the entire study period (11weeks). Biochemical and hematological analysis was supplemented by liver histological examination. Animals treated with the mixture of triterpenic acids exhibited significantly decreased aspartate transaminase and alanine aminotransferase levels and amelioration of the histopathological alterations produced by the anti-TB drugs. The triterpene mixture was able to prevent the steatosis induced by the anti-TB drugs.