Polyherbal Formulations for Diabetes Prof. Dr. Basavaraj K. Nanjwade. M.Pharm., Ph.D. Department of Pharmaceutics KLE University College of Pharmacy Belgaum, Karnataka , India E-mail: bknanjwade@yahoo.co.in Cell No: 00919742431000

14th ACC and ANRAP, Bangkok Graphical Abstract 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Introduction Plant formulation and combined extracts of plants are used a drug of choice rather than individual. Various herbal formulations such as diamed, coagent db, Diasulin, and hyponidd, are well known for their antidiabetic effects. Polyherbal formulation of Annona squamosa and Nigella sativa is composed of medicinal plants (Table 1), which are traditionally used for antidiabetic and antihyperlipidemic activity. The present investigation was undertaken to study the effect of the polyherbal formulation of Annona sqamosa and Nigella sativa on lipidperoxidation and tissue lipid profile in streptozotocin induced diabetic rats. 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Research Background Polyherbal formulation of Annona squamosa and Nigella sativa on blood glucose, plasma insulin, tissue lipid profile, and lipidperoxidation in streptozotocin induced diabetic rats. Aqueous extract of Polyherbal formulation of Annona squamosa and Nigella sativa was administered orally (200 mg/kg body weight) for 30 days. The different doses of Polyherbal formulation on blood glucose and plasma insulin in diabetic rats were studied and the levels of lipid peroxides and tissue lipids were also estimated in streptozotocin induced diabetic rats. The effects were compared with tolbutamide. Treatment with Polyherbal formulation and tolbutamide resulted in a significant reduction of blood glucose and increase in plasma insulin. 7 September 2011 14th ACC and ANRAP, Bangkok

Annona squamosa (Sugar–Apple) 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Nigella sativa 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Material and methods Animals Preparation of drug Chemicals Drug administration Streptozotocin-induced diabetes Experimental design Biochemical analysis 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Experimental design In the experiment, a total of 42 rats (30 diabetic surviving rats, 12 normal rats) were used. The rats were divided into seven groups of six rats each after the induction of streptozotocin diabetes. 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Experimental design Group1: Normal treated rats Group 2:Normal rats given aqueous solution of Polyherbal formulation (200 mg/kg body weight) daily using an intragastric tube for 30 days. Group 3:Diabetic control rats. Group 4: Diabetic rats given aqueous solution of Polyherbal formulation (50 mg/kg body weight) daily using an intragastric tube for 30 days. 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Experimental design Group 5: Diabetic rats given aqueous solution of Polyherbal formulation (100 mg/kg body weight) daily using an intragastric tube for 30 days. Group 6: Diabetic rats given aqueous solution of Polyherbal formulation (200 mg/kg body weight) daily using an intragastric tube for 30 days. Group 7: Diabetic rats given aqueous solution of Tolbutamide (250 mg/kg body weight) daily using an intragastric tube for 30 days. 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Biochemical analysis Estimation of blood glucose and plasma insulin Estimation of lipid peroxidation Estimation of lipids i. Lipids ii. For total cholesterol estimation iii. Fro triglycerides estimation iv. Phopholipids content v. Free fatty acids vi. Statistical analysis 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Table 1: Polyherbal Formulation of Annona Squamosa and Nigella sativa (Composition and Concentration) Sl. No Botanical Name Common Name Family Part used Conc. 1. Annona squamosa Sharifa Annonnacceae Matured fruits 50 2. Nigella sative Kalonji Ranunculaceae seeds 7 September 2011 14th ACC and ANRAP, Bangkok

Fasting blood glucose (mg/dI) Plasma insulin (IU/ml) Table 2: Changes in blood glucose and plasma insulin levels of control and experimental animals Group Fasting blood glucose (mg/dI) Plasma insulin (IU/ml) Normal 81.04 ± 2.29 11.26 ± 0.96 Diabetic control 262.24 ± 22.23 3.48 ± 0.69 Diabetic + Polyherbal formulation (50 mg/kg) 209.58 ± 12.46 5.59 ± 0.34 Diabetic + Polyherbal formulation (100 mg/kg) 155.58 ± 11.69 6.03 ± 0.45 Diabetic + Polyherbal formulation (200 mg/kg) 104.16 ± 6.56 7.15 ± 0.45 Diabetic + Tolbutamide (250 mg/kg) 110.65 ± 9.35 6.32 ± 0.48 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Table 3: Changes in levels of TBARS and Hydroperoxides in liver and kidney of control and experimental animals Groups TBARS Hydroperoxide Liver Kidney Normal 0.58 ± 0.066 0.68 ± 0.05 70.38 ± 4.54 55.34 ± 4.55 Diabetic + Polyherbal formulation (200 mg/kg) 0.56 ± 0.079 0.78 ± 0.06 68.59 ± 5.14 52.48 ± 4.75 Diabetic control 1.54 ± 0.06 1.65 ± 0.12 99.98 ± 5.98 78.29 ± 4.58 0.64 ± 0.053 0.97 ± 0.07 80.55 ± 5.69 60.99 ± 4.78 Diabetic + Tolbutamide (250 mg/kg) 0.67 ± 0.088 1.02 ± 0.08 84.35 ± 5.45 62.45 ± 5.56 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Table 4: Changes in levels of cholestrol, free fatty acids, triglycerides and phospholipids in liver of control and experimental animals ( mg/100g wet. tissue) Groups Cholesterol Free fatty acids Triglycerides Phospholipids Normal 335.44 ± 18.90 606.10 ± 1.76 344.50 ± 23.20 1598.00 ± 19.30 Normal + Polyherbal formulation (200 mg/kg) 328.38 ± 5.74 601.93 ± 9.00 341.10 ± 21.00 1593.10 ± 24.10 Diabetic control 496.56 ± 13.10 921.60 ± 44.60 622.50 ± 18.80 1858.60 ± 18.70 Diabetic + Polyherbal formulation (200mg/kg) 398.65 ± 17.54 769.16 ± 3.30 456.25 ± 17.30 1718.80 ± 14.40 Diabetic + Tolbutamide (250 mg/kg) 405.21 ± 9.79 802.80 ± 3.77 530.80 ± 35.70 1769.30 ± 17.60 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Table 5: Changes in levels of cholestrol, free fatty acids, triglycerides and phospholipids in kidney of control and experimental animals ( mg/100g wet. tissue) Groups Cholesterol Free fatty acids Triglycerides Phospholopids Normal + Polyherbal formulation (200 mg/kg) 372.08±8.28 432.51±1.60 278.75±14.60 1442.50±43.30 Diabetic control 546.90±23.80 743.00±5.70 501.10±34.10 2041.50±33.60 Diabetic+Polyherbal formulation (200 mg/kg) 435.20±12.18 556.80±8.50 382.90±9.28 1684.00±28.80 Diabetic+ Tolbutamide (250 mg/kg) 449.90±13.49 600.30±3.40 438.66±39.30 1819.30±34.70 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Conclusion Polyherbal formulation of Annona squamosa and Nigella sativa, exert a significant antihyperlipidemic. This could be due to combined effect of Annona squamosa and Nigella sativa. Hence the antihyperlipidemic effect of polyherbal formulation of Annona squamosa and Nigella sativa in particular could be considered as of possible therapeutic value 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Liposomes Spherical vesicles with a phospholipid bilayer Hydrophilic Hydrophobic Hydrophobic 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Liposome Preparation 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Liposome Preparation 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Liposome Preparation 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok Lipid Peroxidation Most phospholipid liposomes contain unsaturated acyl chains as part of their molecular structure and susceptible to oxidative degradation. It can be minimized by the use of animal derived lipids like egg PC, which has less saturated lipids, use of light resistant containers, use of antioxidants are useful in minimizing oxidation. 7 September 2011 14th ACC and ANRAP, Bangkok

Diabetic Current Research Mangifera indica(stem,fruits,etc) – Anacardiaceae – Mango Gossypiumherbaceum(flowers,etc ) – Malvaceae – Cotton Cocos nucifera(roots,etc) – Arecaceae – Coconut Lawsonia inermis(bark,etc) – Lythraceae - Mendhi 7 September 2011 14th ACC and ANRAP, Bangkok

14th ACC and ANRAP, Bangkok My Research Group 7 September 2011 14th ACC and ANRAP, Bangkok

THANK YOU E-mail: bknanjwade@yahoo.co.in Cell No: 0091 9742431000 7 September 2011 14th ACC and ANRAP, Bangkok