1. 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.inbknanjwade@yahoo.co.in Cell No: 00919742431000

2. Graphical Abstract 7 September 201114th ACC and ANRAP, Bangkok2

3. 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 201114th ACC and ANRAP, Bangkok3

4. 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 201114th ACC and ANRAP, Bangkok4

5. Annona squamosa (Sugar–Apple) 7 September 201114th ACC and ANRAP, Bangkok5

6. Nigella sativa 7 September 201114th ACC and ANRAP, Bangkok6

7. Material and methods Animals Preparation of drug Chemicals Drug administration Streptozotocin-induced diabetes Experimental design Biochemical analysis 7 September 201114th ACC and ANRAP, Bangkok7

8. 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 201114th ACC and ANRAP, Bangkok8

9. 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 201114th ACC and ANRAP, Bangkok9

10. 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 m g /kg body weight) daily using an intragastric tube for 30 days. 7 September 201114th ACC and ANRAP, Bangkok10

11. 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 201114th ACC and ANRAP, Bangkok11

12. Table 1: Polyherbal Formulation of Annona Squamosa and Nigella sativa (Composition and Concentration) Sl. NoBotanical NameCommon Name FamilyPart used Conc. 1.Annona squamosa SharifaAnnonnacceaeMature d fruits 50 2.Nigella sativeKalonjiRanunculaceaeseeds50 7 September 201114th ACC and ANRAP, Bangkok12

13. GroupFasting blood glucose (mg/dI) Plasma insulin (IU/ml) Normal81.04 ± 2.2911.26 ± 0.96 Diabetic control 262.24 ± 22.233.48 ± 0.69 Diabetic + Polyherbal formulation (50 mg/kg) 209.58 ± 12.465.59 ± 0.34 Diabetic + Polyherbal formulation (100 mg/kg) 155.58 ± 11.696.03 ± 0.45 Diabetic + Polyherbal formulation (200 mg/kg) 104.16 ± 6.567.15 ± 0.45 Diabetic + Tolbutamide (250 mg/kg) 110.65 ± 9.356.32 ± 0.48 7 September 201114th ACC and ANRAP, Bangkok13 Table 2: Changes in blood glucose and plasma insulin levels of control and experimental animals

14. GroupsTBARSHydroperoxide LiverKidneyLiverKidney Normal 0.58 ± 0.0660.68 ± 0.0570.38 ± 4.5455.34 ± 4.55 Diabetic + Polyherbal formulation (200 mg/kg) 0.56 ± 0.0790.78 ± 0.0668.59 ± 5.1452.48 ± 4.75 Diabetic control 1.54 ± 0.061.65 ± 0.1299.98 ± 5.9878.29 ± 4.58 Diabetic + Polyherbal formulation (200 mg/kg) 0.64 ± 0.0530.97 ± 0.0780.55 ± 5.6960.99 ± 4.78 Diabetic + Tolbutamide (250 mg/kg) 0.67 ± 0.0881.02 ± 0.0884.35 ± 5.4562.45 ± 5.56 7 September 201114th ACC and ANRAP, Bangkok14 Table 3: Changes in levels of TBARS and Hydroperoxides in liver and kidney of control and experimental animals

15. Table 4: Changes in levels of cholestrol, free fatty acids, triglycerides and phospholipids in liver of control and experimental animals ( mg/100g wet. tissue) GroupsCholesterolFree fatty acidsTriglyceridesPhospholipids Normal335.44 ± 18.90606.10 ± 1.76344.50 ± 23.201598.00 ± 19.30 Normal + Polyherbal formulation (200 mg/kg) 328.38 ± 5.74601.93 ± 9.00341.10 ± 21.001593.10 ± 24.10 Diabetic control496.56 ± 13.10921.60 ± 44.60622.50 ± 18.801858.60 ± 18.70 Diabetic + Polyherbal formulation (200mg/kg) 398.65 ± 17.54769.16 ± 3.30456.25 ± 17.301718.80 ± 14.40 Diabetic + Tolbutamide (250 mg/kg) 405.21 ± 9.79802.80 ± 3.77530.80 ± 35.701769.30 ± 17.60 7 September 201114th ACC and ANRAP, Bangkok15

16. GroupsCholesterolFree fatty acidsTriglyceridesPhospholopids Normal + Polyherbal formulation (200 mg/kg) 372.08±8.28432.51±1.60278.75±14.601442.50±43.30 Diabetic control 546.90 ±23.80 743.00 ±5.70 501.10 ±34.10 2041.50 ±33.60 Diabetic+Polyher bal 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 201114th ACC and ANRAP, Bangkok16 Table 5: Changes in levels of cholestrol, free fatty acids, triglycerides and phospholipids in kidney of control and experimental animals ( mg/100g wet. tissue)

17. 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 201114th ACC and ANRAP, Bangkok17

18. Liposomes 7 September 201114th ACC and ANRAP, Bangkok18 –Spherical vesicles with a phospholipid bilayer Hydrophilic Hydrophobic

19. 7 September 201114th ACC and ANRAP, Bangkok19

20. Liposome Preparation 7 September 201114th ACC and ANRAP, Bangkok20

21. Liposome Preparation 7 September 201114th ACC and ANRAP, Bangkok21

22. Liposome Preparation 7 September 201114th ACC and ANRAP, Bangkok22

23. 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 201114th ACC and ANRAP, Bangkok23

24. 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 20112414th ACC and ANRAP, Bangkok

25. My Research Group 7 September 201114th ACC and ANRAP, Bangkok25

26. THANK YOU E-mail: bknanjwade@yahoo.co.in Cell No: 0091 9742431000bknanjwade@yahoo.co.in 7 September 201114th ACC and ANRAP, Bangkok26