Design And Insilico Investigation of Novel Kojic Acid Isoniazid Derivatives as Potent Anti Tuberculosis Agents

Tuberculosis: Tuberculosis (TB) is an infectious disease usually caused by the bacterium Mycobacterium tuberculosis (MTB). Tuberculosis generally affects the lungs, but can also affect other parts of the body. Most infections do not have symptoms, in which case it is known as latent tuberculosis.¹ Latent tuberculosis infection (LTBI) means a patient is infected with Mycobacterium tuberculosis, but the patient does not have active tuberculosis. About 10% of latent infections progress to active disease which, if left untreated, kills about half of those infected. ¹ The classic symptoms of active TB are a chronic cough with blood-containing sputum, fever, night sweats, and weight loss. Tuberculosis is spread through the air when people who have active TB in their lungs cough, spit, speak, or sneeze. People with latent TB do not spread the disease. Active infection occurs more often in people with HIV/AIDS and in those who smoke. Diagnosis of active TB is based on chest X-rays, as well as microscopic examination and culture of body fluids. Diagnosis of latent TB relies on the tuberculin skin test (TST) or blood tests. Tuberculosis  (TB) is an infectious disease usually caused by the bacterium Mycobacterium tuberculosis (MTB).² Approximately 8 million new cases and 2 million deaths from tuberculosis occur each year.³ Tuberculosis is caused by a bacterium called mycobacterium tuberculosis. Mycobacterium tuberculosis (Figure No. 1) is slender rod like bacillus, 0.5 micro meter, is neutral on gram staining.

Isoniazid: Numerous functions of the pyridine ring as a heterocyclic moiety have been discovered. These can be found in many different kinds of medications. Isoniazid (INH) is one such pyridine-containing substance. INH is known by its chemical name, isonicotinic acid hydrazide. An antibiotic called isoniazid (INH) is recommended as a first line of treatment for active Mycobacterium tuberculosis infections. Isoniazid is a first-line drug used in active tuberculosis (TB) treatment since 1952,⁴ The only reason why isoniazid (INH) derivatives need to be synthesized is because Mycobacterium tuberculosis bacteria are now resistant to both isoniazid and several other first-line medications, including pyrazinamide, ethambutol (EMB), and rifampicin (RIF).⁵ Isoniazid possesses multiple characteristics, including antimycobacterial, antibacterial, anti-virus, antimicrobial, antimalarial, and anti-convulsant effects, in addition to its ability to prevent tuberculosis. Isoniazid is a first-line drug used in active tuberculosis (TB) treatment since 1952,⁴ Isoniazid use has been linked to a number of adverse drug reactions (ADRs), including hepatotoxicity, peripheral neuropathy, gastrointestinal issues such nausea and vomiting, psychosis, cutaneous adverse drug reactions (CADRs), and pancreatitis.⁶ It was Meller and Malley that originally identified this synthetic medicine in 1912.⁶

Pharmacokinetics:

Absorption: After administration by mouth or intramuscular (IM), there is a quick and thorough absorption. One to two hours is when the plasma concentration peaks. INH is taken up by the small intestine and then made its way to the liver through the portal system.

Distribution: INH is rapidly distributed into all body tissue compartments, including cerebrospinal fluid. Plasma protein binding is 10% to 15%.

Metabolism: N-acetyltransferase 2 (NAT2) in the liver and intestines acetylates INH to render it inactive. Acetyl hydrazine is oxidized by CYP2E1 into hepatotoxic metabolites once INH is acetylated. Because they can oxidize more quickly, people who metabolize quickly may produce more hepatotoxins. Against hepatotoxins, glutathione S-transferase (GST) offers protection. When GST is combined with hazardous metabolites, the resulting metabolites are largely water soluble, comparatively non-toxic, and quickly eliminated.

Elimination: Saliva and faces excrete tiny amounts of the drug, while urine excretes the majority of the medication (75–95%) and its metabolites.⁶

Adverse Effect: NH use has been linked to a number of negative consequences, the majority of which are transitory and low grade. Most patients report gastrointestinal side effects, although some also suffer pruritus and redness. A common side effect of INH therapy is peripheral neuropathy, but with an incidence of less than 0.2%. The risk may increase in patients with diabetes, HIV, nutritional deficiency, renal failure, alcoholism, and in women who are pregnant or breastfeeding.^8,9 Drug-induced lupus erythematosus (DILE) has been reported in up to 1% of patients taking INH.¹⁰

Kojic Acid: Kojic acid is a fungal metabolic product produced by a few species of aspergillus, especially by A.oryzae, which has the Japanese common name koji. This acid is a by product in the fermentation process of malting rice, for use in the manufacturing of sake, the Japanese rice wine. From the mycelium of the fungus Aspergillus oryzae cultivated on steamed rice, Saito discovered kojic acid in 1907. Kojic acid orginally named as koji which is the name of fungus from which kojic acid derived. Yabuta then gave the substance the name kojic acid in 1913. Additionally, Yabuta clarified the structure of kojic acid. Kojic acid can produced by variety of microorganisms under various aerobic condition by various carbohydrate source and lethal dose in living things is about 1g/kg. Since the early twentieth century, it has been known as an additive to prevent browning of food material such a crab, shrimp and fresh vegetable in food industry (e.g., as an antioxidant or anti browning agent) in order to preserve their freshness and to inhibit discoloration.³² It play an important role in iron overload disease such as beta thalassemia or anaemia, since it possess iron chelating activity, also it form stable complexes of metal kojate via reaction of kojic acid with acetate salt such as tin, beryllium, zinc, copper, nickel, cobalt, iron, manganese, chromium, gold, palladium, indium, gallium, vanadium, aluminium. Kojic acid is a well-known whitening and lightening ingredient that is widely used in cosmetic and skin care formulations. Its processes include inhibiting the formation of melanin, which contributes to the whitening of the skin. The market for kojic acid has existed for almost 40 years, starting in 1955. charles Pfizer & American companies announced their 1^st try to produce its organic acid. ¹¹

Experimental Work:

Draw the scheme diagram using chemdraw software or chemsketch. Specific atom given the specific colour for easy indication.

Estimation of Lipinski’s rule of 15 synthetic compound using molinspiration software or also used swissADME software. Evaluation of drug likness and pharmacokinetic parameter using swissADME Software. Estimation toxicity profile using data warrior software. Estimation of binding energy of newly synthesis koalniazid molecules using autodock software. Resulting binding energy of 25 koalniazid compound compare with standard isoniazid drug.

Lipinski rules of five: the exception of one compounds, all 15 compounds comply with the Lipinski rules for analysis. there are, Since molecule number 4 (N-((5-Hhydroxy-2-(hydroxymethyl)-4-oxo-4H-pyran-3-yl)(3-nitrophenyl)methyl)isonicotinohydrazide) has a hydrogen bond acceptor value of greater than 10, it does not meet the third Lipinski rules analytical criteria for high bioavailability. According to the lipinski rules of analysis only one compound not good bioavailability remining compound is obey the lipinski rules of analysis so that compound is good bioactivity score. (Table No. 2)

Toxicity profile: Isoniazid molecules are a common medicine used to treat tuberculosis, but they have a severe toxicity profile that includes mutagenesis, tumerogenic, reproductive toxicity, and irritation. The newelly design 15 compounds overcomes this drawback.  some of them lower toxicity profile compare to standard drug of isoniazid. Isoniazid contain hydrazine molecules responsible for toxicity of compounds, various derivatives added to the  molecules decrease toxicity profile. (Table No. 3)

Molecular Docking analysis: To ensure the interaction between the synthetic compounds and tuberculosis disorder associated targets, we performed molecular docking analysis using autodock 4.2.7. each of the compounds was docked with enoyl reductase target individually, the output of all ligands was given by energy values in kcal/mol as shown table. These compounds showed very good binding affinity with target enoyl reductase receptor when compared to standard drugs (isoniazid).The docking score of the compounds targeted enoyl reductase enzyme receptor was comparted with the score of the drug isoniazid which is used as a potent drug for tuberculosis disorder. Table show the result obtained through the docking study between the 15 compound with enoyl reductase . that all compound involved in this study interacted with the enoyl reductase attractively when compared with standard drug, and the compound molecules 10,18 obtained lower interaction energy. Being shown to be more stable in complex with the site of the macromolecules. Molecule 2 more interaction compare to standard drug of isoniazid and also stable conformation with molecules 2 at the active site of enoyl reductase. (Table No. 4)

Table No. 2

Table No. 3

Table No. 4