Studying the effects of a novel drug (NS-009) in the treatment of diabetes using animal models.

Abstract

Being consistently increasing around the world, diabetes requires special attention in the development of effective novel drugs having more potent and posing least side effects. Since the reporting of many diabetes drugs in the involvement of cancer and other disease implications, it became inevitable to test the efficacy of new drugs. Therefore, this study focused on the effects of a novel drug NS-009 on lowering the levels of both glucose and insulin in an experimental design utilizing Sprague-dawley rat models of diabetes. The study found the mean blood glucose values of 104 + 2.01, 229 + 3.93, and 125 + 6.31 mg/dL in and mean blood insulin levels of 6.83 + 0.65, 22.8 + 1.92, and 7.00 + 0.97 μU/mL in normal control, disease control and treatment groups, respectively. This is the first study of its kind demonstrating the dual effects of normalizing both the blood glucose and insulin levels.

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Introduction

A collection of metabolic disorders in which hyperglycemia stands out as their characteristic feature is generally known as the diabetes mellitus. The most common causes leading to this hyperglycemic blood levels include defective insulin secretion, its absorption and action in the cell, or sometime both of these. The long term consequences of the diabetic chronic hyperglycemia include initially damage and dysfunction which ultimately can lead to the failure, if uncontrolled, of various organs such as kidneys, heart, blood vessels, eyes and nerves (Unnikrishnan et al., 2016, Nathan, 2015).

Hyperglycemia can also be identified with some of its most obvious symptoms which include blurred vision, sudden weight loss or gain, polyphagia, polydipsia and polyuria. In addition to these, growth impairment also occurs in some cases while in others there is an increases susceptibility of various infections along with slower healing times of the body. There are two major types of diabetes – type 1 and type 2 diabetes – with the former known by the β-cells’ destruction while the latter being characterized by the insulin resistance of the tissues as well as its secretion deficits (American Diabetes, 2009).

Diabetes development in a person includes activity of several pathogenic processes. The most common of these is the destruction of pancreatic β-cells via the action of autoimmune cells on them. Consequently, there is deficiency of insulin in the body as the main source of insulin production is destroyed. In other instances of diabetes, some abnormal body processes result in an overall increased resistance of the body to the insulin, thereby creating an insulin deficiency situation in the disease condition despite the availability of the insulin. This inadequacy of insulin in either way, results in the anomalous metabolism of the tissue carbohydrate, protein and fat metabolism as well. Also, it is a common occurrence of both the diminished insulin production along with its increases tissue resistance in a same patient, in this manner making the identification of the primary cause of diabetes even harder (Wilcox, 2005, Cerf, 2013).

Although may drugs are currently available and in use by the public for the management of diabetes, still there is always a need for the development and identification of novel drugs so as to find more efficient ones among all. Therefore, the current study was designed to find the effects of a novel drug NS-009 on the production of glucose and insulin using rat models. The rat models have been used for this study as they share 90% genome identity with the human genome and thus can serve as appropriate substitutive models for human studies before clinical trial can begin.

Material and methods

Before starting the experiment on animal models of diabetes, an approval was taken from the Animal Ethics Committee of Australia. The experiment was designed to be completed within a timeframe of 4 weeks.

Animal selection and maintenance

A total number of 18 sprague-dawley rats were taken and divided into 3 groups of 6 rats each. Also, the rats taken for this study were all of same gender i.e., males, to avoid gender based differences which might arise in the results and hence may create ambiguity.

The animals were kept in cages with each cage representing 1 group of 6 rats. An appropriate bedding was provided for the rats with food and water available ad libitum. This provision of food and water was maintained only for the first week of the experiment. Also, the rats were kept in 12 hours light dark cycle with environmental temperatures maintained to around 22 + or – 1 °C.

Experimental groups and diet

During the first week of the experiment, all three groups of animals were given the normal diet followed by high fat diet to groups 2 and 3 during the 2^nd week while maintain the normal diet for the first group which is serving as a control group for this experimental design. In the final weeks of the experiment i.e., 3^rd and 4^th weeks, the group 4 of rats was given the treatment with the novel drug NS-009. In this way, the division of the animal groups is as follows: group 1 – control group; group 2 – diabetic type group; and group 3 – NS-009 treatment group with diabetes type.

Collection of blood samples

The blood samples from the rat tails were collected following the standard protocol. Briefly, the tails were immersed in a water bath with temperature maintained at 42 °C so as to dilate the blood vessels and help locate the tail vein. After this, the tail was dried with a clean towel and punctured with a syringe creating negative pressure. Then the blood was drawn by keeping the tail straight till the blood sample was collected. After collecting adequate sample, the syringe was removed carefully and a bandage with antiseptic solution was applied with a slight pressure to the tail to stop bleeding.

Statistical analysis

Of all the values collected for the serum biomarkers – glucose and insulin – mean values of the individual groups were calculated along with their respective standard errors. Moreover, the box plots of the values were plotted for all three groups with respect to each biomarker for a better visualization of the differences observed between control groups, diabetes groups and treatment groups.

Results

The results of the experiment are given in the table 1 and visually represented in the form of box plots in the Figure. 1. The mean glucose level values in the control group were 104 + 2.01 (mg/dL) which increased to 229 + 3.93 (mg/dL) in the diabetic group. However, in the third group – the diabetic mice with drug treatment group – the glucose mean values were again decreased to 125 + 6.31 (mg/dL) (Table 1 and Fig. 1 (a)).

About the insulin levels, the mean values came out to be 6.83 + 0.65 (μU/mL) in the control group; 22.8 + 1.92 (μU/mL) in the diabetes group and finally 7.00 + 0.97 (μU/mL) in the diabetic treatment group (Table 1 and Fig. 1 (b))