Computational Studies and Synthesis of few Thiazolidine-2,4-Dion Analogues for Peroxisome Proliferator Activator Receptor- γ Agonist as Useful Antidiabetic Agent

 

Shital N. Chaskar*, Dr. Sanjay J. Kshirsagar

Department of Pharmaceutical Chemistry, MET’s Institute of Pharmacy, Nasik, Maharashtra, India.

*Corresponding Author E-mail: shitalchaskar7@gmail.com

 

ABSTRACT:

Peroxisome proliferator activated receptor (PPAR)-γ is a member of the nuclear hormone receptor super family and the molecular target for the Thiazolidinediones (TZD), used clinically to treat insulin resistance in patients with type 2 diabetes. In addition to their ecacy to improve insulin sensitivity. Diabetes mellitus is a metabolic disease characterized by the presence of chronic hyperglycemia accompanied by greater or lesser impairment in the metabolism of carbohydrates, proteins and lipids. The present work shows the rational design of some novel PPAR-γ agonists involving computational study performed by Molecular Design Suite (MDS) in to ligand binding domain of PPAR-γ receptor to explore conformations of molecules. The compounds synthesized using multi-step synthesis protocol. The purity of synthesized compound was ascertained by IR, NMR, Mass and elemental analysis and tested by oral glucose tolerance test (OGTT). Two molecules showed most prominent activity on hyperglycemic control.

 

KEY WORDS: Diabetic Mellitus, Thiazolidinedione, Computational study.

 


1. INTRODUCTION:

Diabetes Mellitus is one of the fastest growing disease according to International Diabetes Federation report of 2011 an estimated 366 million people had DM, by 2030 this number is estimated to almost around 552 million. Its incidence is increasing rapidly, and by 2030 this number is estimated to almost around 555 million (Baynes J et al, 2015). Diabetes mellitus is a metabolic disease characterized by the presence of chronic hyperglycemia accompanied by greater or lesser impairment in the metabolism of carbohydrates, proteins and lipids.

 

The effect of these includes long term damage, dysfunction and failure of various vital organs (Rang HP et al, 2007). TZD class of oral agents for treatment of type 2 diabetes mellitus. It increases insulin sensitivity by affecting the peroxisome proliferators activated receptor-ɣ (PPAR-ɣ). Acting as an agonist to this receptor, they decrease insulin resistance in adipose tissue, skeletal muscles and liver (Mark S, 2003).

 

TZDs, such as rosiglitazone, pioglitazone, troglitazone, and ciglitazone are insulin-sensitizing drugs and have constituted a major advance in the recent therapeutic management of T2DM. In addition to improving insulin sensitivity, TZDs have also effects on TG, FFA, and ketone body level in several animal models of T2DM. Current medications are effective in maintaining hyperglycemia. Diet, exercise and careful monitoring of blood glucose level are the keys to manage diabetes so that patient can live a relatively normal life (A Ramachandran et al). There is an urgent need for novel anti-diabetic agents that should have efficacy with potential to reduce complications.

2. MATERIAL AND METHODS:

2.1.  Instrumentation:

All chemicals used for synthesis were purchased from Sigma-Aldrich and Merck. Melting point was measured on melting point apparatus. The infrared spectra were recorded in KBr discs on win IR FTS 135 instrument. The 1H NMR spectra were recorded by Sigma-Aldrich as internal standard. Mass spectra were obtained on JEOL-JMS-DX 303 instrument. Thin layer chromatography was carried out using silica gel G-coated TLC plates.

 

2.2.  QSAR:

Quantitative structure-activity relationship (QSAR) methodology was introduced by Hansch et al. in the early 1960s. The approach stemmed from linear free-energy relationships in general and the Hammett equation in particular. It is based on the assumption that the difference in structural properties accounts for the difference in biological activities of compounds. According to this approach, the structural changes that affect the biological activities of a set of congeners are of three major types: electronic, steric, and hydrophobic. These structural properties are often described by Hammett electronic constants, Verloop STERIMOL parameters, hydrophobic constants, etc. The quantitative relationship between a biological activity (or chemical property) and the structural parameters could be conventionally obtained using multiple linear regression (MLR) analysis. Many different approaches to QSAR have been developed since Hansch’s seminal work. These include both 2D (two-dimensional) and 3D (three-dimensional) QSAR methods. The major differences of these methods can be analyzed from two viewpoints: (1) the structural parameters that are used to characterize molecular identities and (2) the mathematical procedure that is employed to obtain the quantitative relationship between a biological activity and the structural parameters.

 

2.3.  Synthesis of target compounds:

2.3.1.       Synthesis of 2,4-thiazolidinedione:

In a 250ml three-necked flask, a solution containing 56.4g(0.6M) of Chloroacetic acid in 60ml of water and 45.6g(0.6M) of Thiourea was dissolved in 60ml of water. The mixture was stirred for 15minute till occurrence of white precipitates. To contents of flask was now added slowly 60ml of conc. hydrochloric acid from dropping funnel to dissolve the precipitates, after which the reaction mixture was stirred and refluxed for 16-17hrs at 100-110ºC, on cooling the contents of flask were solidified to a mass of clusters of white needles. The product was filtered and washed with water to remove traces of hydrochloric acid and dried. It was purified by recrystallization from ethyl alcohol. The yield was 76%.

2.3.2.       Synthesis of 5-Substituted 2,4-thiazolidinediones:

To solution of P-hydroxy benzaldehyde (0.25M) and Compound A(0.25M) in hot glacial acetic acid (50ml), fused sodium acetate (1.8g) was added and then it was refluxed for 6-7hrs with occasional shaking. It was poured in cold water, then product obtained was filtered, wash with water, alcohol and ether. Product was recrystallised from glacial acetic acid to obtain yellowish brown needle like crystals. The yield was 69%.

 

2.3.3.       Synthesis of Synthesis of 4-(2,4-dioxothiazolidin-5-ylidene)methyl)phenyl benzoate:

In a conical flask containing 0.005M(1.12gm) of Compound B (5-(4-hydroxybenzylidene)thiazolidine-2,4-dione) in which few ml NaOH Solution is added just to dissolve the compound. In this solution add dropwise Benzoyl Chloride 0.005M(0.8ml), stirred it well up to 15 minutes. Product which separated was collected under pressure by suction pump and dried. It was then recrystallized from ethanol to yield crystals.

 

2.3.4.       Synthesis of 4-(2,4-dioxothiazolidin-5-ylidene)methyl)phenyl-2-chlorobenzoate:

In a conical flask containing 0.005M (1.12gm) of Compound B (5-(4-hydroxybenzylidene) thiazolidine-2,4-dione) in which few ml NaOH Solution is added just to dissolve the compound. In this solution add dropwise 2- Chloro Benzoyl Chloride 0.005M (0.8ml), stirred it well up to 15 minutes. Product which separated was collected under pressure by suction pump and dried. It was then recrystallized from ethanol to yield crystals.

 

2.3.5.       Synthesis of 4-(2,4-dioxothiazolidin-5-ylidene)methyl)phenyl-4-chlorobenzoate:

In a conical flask containing 0.005M (1.12gm) of Compound B (5-(4-hydroxybenzylidene) thiazolidine-2,4-dione) in which few ml NaOH Solution is added just to dissolve the compound. In this solution add drop wise 4- Chloro Benzoyl Chloride 0.005M (0.8ml), stirred it well up to 15 minutes. Product which separated was collected under pressure by suction pump and dried. It was then recrystallized from ethanol to yield crystals.

 

2.3.6.       Synthesis of 4-(2,4-dioxothiazolidin-5-ylidene)methyl)phenyl-3-bromobenzoate:

In a conical flask containing 0.005M (1.12gm) of Compound B (5-(4-hydroxybenzylidene) thiazolidine-2,4-dione) in which few ml NaOH Solution is added just to dissolve the compound. In this solution add drop wise 3- Bromo Benzoyl Chloride 0.005M (0.8ml), stirred it well up to 15 minutes. Product which separated was collected under pressure by suction pump and dried. It was then recrystallized from ethanol to yield crystals.


 

 

 


3.      RESULT AND DISCUSION:

3.1.  Design of PPAR-γ agonist:

Thiazolidinedione is hypoglycemic agents which are structurally compose of an acidic head group connected to lipophilic Tail by a phenoxyalkyl linker.

 

 

Fig. 1. Pharmacophore of PPAR-γ agonist.

 

3.2.  Chemistry

The IR spectra of synthesized TZD molecules. The primary amine was characterized by peaks in range 3300-3500 cm-1, NH bending was observed in range 1600-1650 cm-1. The 1H NMR shows the prominent structural features. The mass spectra described the base peaks having exact molecular mass with fragment peaks.

 

3.3.  In vivo estimation of Blood glucose level:

3.3.1.       Oral glucose tolerance test (OGTT):

Healthy wistar rats of both sexes weighing 200 to 250 gm included in this study. Animals were randomly divided into three groups, five animals in each group.

Group I: Vehicle control (normal saline).

Group II: Diabetic control (Glucose 2g/Kg).

Group III: Diabetic + MCR-3-15/17 and MCR-4-15/17 [test control]

Rats were dosed following an fast for 6 hrs. 2 gm/kg glucose administered to each rat of glucose control group and blood samples were collected from rat tails and the glucose level was checked using blood glucose meter after 0, 15, 30, 60, 120 min. The Plain drug suspension prepared by using DMSO (Dimethyl sulfoxide) and glucose level were checked. The experiment was again repeated by administering samples containing glucose and optimized synthesized derivatives in the same dose to test control.

 

TABLE.1. Hypoglycemic effects of synthesized test compounds

Treatment

Blood Glucose Level (mg/dl)

30min

60min

120min

Pioglitazone

142±3.44

96.25±6.98

90.25±1.31

MCR-011

136±1.95

113.5±2.21

102±1.58

MCR-012

131±1.68

117.5±1.70

107.75±3.01

Glucose control

150.5±14.26

149±0.70

131±2.17

Saline control

119±2.27

116.5±2.39

114±2.16

Values are expressed as mean ± S.E.M

 

The pharmacological evaluation of the compounds showed increase in blood glucose level. Decrease in the blood glucose level compared to control. The compounds MCR-003-16/17 and MCR-004-15/17 showed highest percentage decrease of blood glucose level at the dose of 1/10th that of LD50 among the evaluated compounds compared to control. The analysis of structural features revealed that substitution of thiazolidinedione group enhanced the hypoglycemic potential of the synthesized compounds.

 

4.      CONCLUSION:

Most drugs are available and approved currently but cannot reach an adequate level of glycemic control in diabetic patients, and have many side effects, thus there is urgent need of newer agents with the potential to maintain normal glucose level along with better tolerance. There are different drugs are available in the market for the treatment of diabetes mellitus, not a single drug, they are used in combination with insulin is effective in maintaining blood glucose (HbA1C) levels below 7 as recommended by American Diabetes Association.

 

Four compounds were synthesized by using three steps, in first step thiazolidinedion was synthesized, in step second by using these thiazolidinedione, 5-(4-substituted benzylidine)-2,4-thiazolidinedion derivatives were synthesized. Then analyses of compounds were performed by melting point, Thin-layer chromatography, FT-IR, NMR, Mass spectroscopy and elemental analysis. Biological evaluation was performed by Oral glucose tolerance test.

 

5. REFERNCES:

1.       Baynes J Diabetes Metab (2015) Volume 6, Journal of diabetes and metabolism.

2.       Rang HP, Dale MM, Ritter JM, Moore PK.(2007),rang and dale pharmacology china: Churchill livingstone.6th edition p.402-408

3.       Mark S. (2003), Insulin and hypoglycemic agents, Abraham DJ. Bergers medicinal chemistry and drug discovery. Vol.4 6th edition. united states of America :john wiley and sons inc p.1-37

4.       Mcgrady, Angele, James H. (1999), Coplementary/Alternative therapies in General medicine: diabetes mellitus.john WS and Joseph jj. Comple/alterna medicine: an evidence based approach.st.louis,mosby

5.       Donald McClain, Animal Models of Diabetic Complications Consortium Intraperitoneal Glucose Tolerance Testing (IPGTT) Version: 1.0

 

 

 

 

 

 

 

 

Received on 03.07.2017       Accepted on 21.09.2017     

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Res. 2017; 7(4): 265-268.

DOI:   10.5958/2231-5691.2017.00042.9