PROTOCOL STUDY TO EXTRACT MANGIFERA INDICA L. POWDER , (MANGIFERIN PHYTOCONSTINUENT) AND MANGIFERA INDICA BARK OIL AND ITS ANTIDIABETIC, HEPATOPROTECTIVE, IMMUNOMODULATORY AND ANTICANCER ACTIVITY IN EXPERIMENTAL ANIMALS AND TRASGENIC NUDE MICE

   ABSTRACT

  Herbal antidiabetic preparations are often used as an add-on therapy in diabetics. Mangiferin 

indica (Anacardiaceae) stem bark contains a rich content of Mangiferin and is used traditionally 

in Indian Ayurvedic system to treat diabetes. Mangiferin has been reported to be potent AlphaGlucosidase inhibitor that also shows modulation of CYP450 enzyme that inhibits CYP3A4 

enzyme. Pioglitazone, a Thiazolidinediones derivatives decreases insulin resistance via its action 

at the peroxisome proliferator activated receptor subtype gamma (PPARγ) and emerged as a 

novel oral Antidiabetic agent in recent past. It is metabolised by multiple Cytochrome P450 

(CYP) isoenzymes, mainly by CYP2C8, CYP3A4 and CYP2C9. Hence it is speculated that 

Mangiferin may influence the kinetic, diabetic and Hepatotoxicity effect of pioglitazone which is 

particularly crucial, as any increment in its plasma level may raise safety concerns. 

Pharmacokinetic study was carried out by administrating Pioglitazone orally (10mg/kg) alone 

and in combination with Mangiferin (200mg/kg) pretreated in alloxan induced diabetic rats and 

its plasma levels were determined at various time points (0h, 1h, 2h, 4h, 8h, 12h, 18h, 24h) after 

oral administration by HPLC. Mangiferin pretreatment increased AUC0-∞ of pioglitazone. 

Pharmacodynamic effects of Pioglitazone with Mangiferin were evaluated in alloxan induced 

diabetic rats. The blood samples were collected from diabetic rats at different time intervals up to 

24hours and blood glucose was estimated. Mangiferin (200mg/kg, p.o) pretreatment has 

significantly altered the onset of Antidiabetic effect of Pioglitazone from 26.67 % to 31.5% and 

significantly enhanced the peak Antidiabetic effect from 69.25% to 73.99%. Duration of 

Antidiabetic effect was raised from more than 24hrs. 

Hepatotoxicity activity was estimated and Mangiferin significantly reduces the level of AST and 

ALT enzymes alone and in combination with pioglitazone. 

The study indicates that Therapeutic drug monitoring has to be required to readjust the 

therapeutic dose of Mangiferin and pioglitazone when they used concomitantly. 

Beside this oil fraction called mangifera indica bark oil obtained from pet ether fraction 

OF mangifera indica bark has anti cancer activity in nude mice (transgenic mice) to be studied.

Key Words: Mangiferin, Pioglitazone, Alloxan, mangifera indica bark oil Pharmacokinetic, Pharmacodynamic, Transgenic mice

Isolation, purification and characterization of Mangiferin from Mangifera indica L. stem barks^128,129.

Mangifera indica stem barks were collected from the Kyarkop region of Dharwad (Karnataka, India) in the month of September 2010. The stem barks were inspected to be healthy, collected and were washed with water thrice in the bucket and dried overnight over a cotton cloth. After that the materials were grounded to fine powder using a domestic electric grinder. Powdered stem barks were passes through the sieve no.20 and around 100 gram of powder was successively extracted with petroleum ether (2× 300ml; 4 hr each time) and acetone(400 ml 4 hr each time) soaked for 24hours to remove  tannins. Finally bark extracted with 70% ethanolic

solvent (2×400 ml; 4hr each time). The ethanolic fraction was concentrated using distillation unit and evaporated on water bath. Then mangiferin was separated as a yellow colour powder . Confirmative tests for isolated mangiferin were performed by dissolving isolated mangiferin in aqueous ethanol showed deep yellow colour solution due to presence of mangiferin. In addition, alcoholic solution of isolated mangiferin when treated with ferric chloride gave deep green colour while with ferric chloride and hydrochloric acid it produced greenish yellow precipitate immediately due to presence of mangiferin ^.

v     Purification of Mangiferin by crystallization¹³⁰

The Mangiferin obtained is dissolved in 70 % of ethanol and heated in the water bath for 30 minutes and filtered, the filtrate obtained is again reheated for a suitable period for super saturation and then cooled at room temperature. The yellow powder of Mangiferin was obtained after the complete evaporation of the solvent.

v    Determination of Melting Point ^129.

Small amount of Mangiferin was taken in a melting point capillary tube and placed in the melting point apparatus (make) and then it was switched on and the temperature was gradually increased. At certain temperature range the Mangiferin was melted and this temperature was notes as melting point of Mangiferin.

MARS- MAHATMA RAKESH SINGH