Identification of Alpha-glucosidase as a potential enzyme for the treatment of diabetes mellitus 2 by In-Silico approach


1.    INTRODUCTION

Diabetes mellitus is the disease of ineffectual control of blood vessels of glucose. Type 2 diabetes mellitus (Type 2 DM) is a rapidly increasing world health problem referred to as adult-onset diabetes. It is called adult-onset as the patients are diagnosed later in life with this disease. The symptoms appear late making the disease unidentifiable unless around the age of 45. According to Raelic (2016) Type 2 DM is considered to be the disease of middle or older aged people but with the increase of obesity and unhealthy lifestyle Type 2 DM is increasingly being diagnosed at a younger age (Rahelić, 2016). On the onset of Type 2 DM insulin feedback is diminished and this is called insulin resistance. About 3000 years ago an Egyptian manuscript first discussed insulin resistance. Type 2 DM is a multifactorial disease and develops from the presence of several genetic, environmental, and behavioral factors. In other words, the interaction of different factors contributes to the development of Type 2 DM. People having Type 2 DM are more susceptible to different health complications including heart disease and stroke. This study aims to identify alpha-glucosidase as a potential enzyme for the treatment of diabetes mellitus type 2 disease.

2.    Key Words

Type 2 Diabetes Mellitus, Alpha-Glucosidase, Molecular Docking, Auto Dock Vina, In-Silico Drug Identification, phytomedicines

3.    Research Questions

1.      Is Alpha Glucosidase act as a potential enzyme for the treatment of Diabetes mellitus?

2.      Is it possible to delay carbohydrate absorption from the small intestine using Alpha-glucosidase inhibitors (AGIs)?

3.      Is there a potential in plant terpenoids to be used as inhibitors for pancreatic amylase?

4.      Is it possible to reduce obesity and heart diseases in the world by treating Diabetes Mellitus Type 2?

5.      Can Diabetes Mellitus Type 2 be treated by inhibiting alpha-glucosidase without disturbing the bacterial flora of the human body?

 

4.    Objectives

1.      To virtually screen, a library of about 200 plant terpenoids against alpha-amylase.

2.      To utilize the potential of alpha-glucosidase for the discovery of potent drugs against diabetes mellitus type 2

3.      To indirectly treat obesity and heart diseases by treating diabetes mellitus type 2.

4.      To repurpose already existing plant terpenoids for the wellbeing of diabetes mellitus type 2 patients.

5.    Motivation of Research

According to research by Rahelic (2016)  with his colleagues examined about a million people and submitted a systematic review and meta-analysis. They concluded that the younger age diabetics are more to microvascular and macrovascular complications and they are associated with an increased risk of mortality (Rahelić, 2016).

According to World Health Organization, T2DM was the 9th leading cause of death in 2019 (Diabetes, n.d.). Globally in 2021, 462 million individuals are suffering from T2DM and it counts for 6.28% of the world's population. According to Ralph A. DeFronzo, the rate of Type 2 DM will be increased to 591.9 million by 2023, which counts to a 55% increase in Type 2 DM between 2013 to 2023 (DeFronzo et al., n.d.). There is currently no cure for T2DM and its prevalence is increasing rapidly throughout the world causing a high rate of mortality and morbidity particularly in youngsters. Worldwide around 8.4% of deaths are caused by Type 2 DM and its related microvascular or macrovascular complications, as stated by Rahelic in 2016 (Rahelić, 2016).

According to a report published by Narayan and colleagues in 2020 (in an epidemiological study) that South Asians are more susceptible to T2DM than other ethnic groups even at low BMI and they count for almost 2 billion people on the Earth. Further Narayan and his colleagues added that normal-weight south Asians are at more risk to get Type 2 DM as compared to normal-weight people of other ethnic groups. (Narayan & Kanaya, 2020). Until now there is no definite treatment for Type 2 DM or the metabolic dysfunction associated with it. Therefore, there is an urgent need to unknot the cure for T2DM to upgrade the lifestyle of T2DM patients.

6.    Research Contribution

Several papers have been published that describe the effects of Alpha Glucosidase inhibitors in type 2 diabetes, this research will contribute to identifying more alpha-glucosidase inhibitors as an effective way to control diabetes type 2. According to Dirir and his colleagues (2021), there are just three approved inhibitors for alpha-glucosidase as a treatment for Type 2 DM which include miglitol, acarbose, and voglibose. All of them have their limitations, gastrointestinal side effects and none of them can be considered as a promising treatment as Type 2 DM is related to several other health complications and each patient should be prescribed the treatment according to the severity of microvascular and macrovascular complications. (Dirir et al., 2021a) There is a need to find new and potent inhibitors for alpha-glucosidase and this research will contribute to the fulfillment of this need. Academia will get advancement to the knowledge by adding new knowledge through this research. Through this research pharmaceutical industry can contribute to the well-being of society by finding a plant-derived medicine for the treatment of the ninth most common disease in the world (Type 2 DM).

7.    Literature Review

Diabetes mellitus can be treated by slowing starch degradation in the intestine by using Alpha-glucosidase inhibitors. According to Bashary et al (2019) type 2 diabetes is linked to diminished glucose tolerance due to insulin resistance. Any injury in the islets of beta cells of the pancreas can cause insulin deficiency that will greatly affect the consumption of glucose by the liver, muscles, and adipose tissues which results in glucose intolerance. Several other genetic, environmental and living conditions also contribute to the development of insulin resistance. (Bashary et al., 2019).

7.1.    Type 2 Diabetes Mellitus

According to Halim et al, the chronic Hyperglycemia condition that results in a group of metabolic disorders is called diabetes and this hyperglycemic condition is due to the deficiency in the levels of insulin. In other words patients with type 2 diabetes mellitus reveal aberrant insulin secretion and insulin action. Genetic, environmental, and some other factors also contribute to T2DM. The main feature of type2 diabetes includes the lack of demand for insulin which is obligatory for the prevention of ketoacidosis. . These impediments can generate substantial influence on the organ system of the body. (Halim et al., n.d.). According to Oguntibeju (2019), these complications are divided into two groups namely vascular complications and non-vascular complications. There are two types of vascular complications, one is microvascular and the other one is macro complications. Neuropathy, nephropathy, and retinopathy are microvascular complications whereas, macrovascular complications include cerebrovascular disease (CVA), coronary artery disease (CAD), coronary heart disease, and peripheral vascular disease (PVD). Additionally, the researcher also suggested that the possibility that an individual will develop cardiovascular diseases naturally increases with the presence of diabetes (Halim et al., n.d.) (Oguntibeju, 2019)

The popular non-insulin treatment for Type 2 DM is modulation of pathophysiological abnormalities that relate to an increase in glucose concentration in blood (hyperglycemia).

7.1.1        Beta Cell Physiology in Type 2 DM:

According to Al-Mrabeh, the beta cells are the part of the pancreas which is the largest gland of the body. Beta cells secrete insulin and along with insulin pancreas participate in the production of glucagon through alpha cells and somatostatin through gamma cells. Beta cells are stimulated through hormonal productions and they can also be stimulated through non-hormonal productions like zinc and ATP. In diabetic patients, the amount of beta cells in the pancreas is found to be lowest than non-diabetics (Biomedicines & 2021, n.d.)

7.1.2.      Insulin Secretion:

In 2018 while talking about the mechanism of insulin action Petersen and Shulman stated that beta-cells of the pancreas sense the output from neurotransmitters, hormones, and different substrate concentrations (e.g. Glucose and amino acids like arginine) and maintain the plasma concentration of insulin accordingly. A non-diabetic person may only need a 0.2 U of insulin for 2 hours in a daily working routine, while an obese and insulin-resistant person will require 45 U of insulin for 2 hours in a daily working routine. This difference in required plasma insulin concentration depicts the importance of beta-cell secretion in a normal amount. Moreover, if insulin secretion is impaired by beta-cell dysfunctioning, it may lead to hyperglycemia which contributes to Type 2 diabetes (Petersen & Shulman, 2018).

7.1.3.      Insulin Resistance:

According to the latest review paper by Courtney and Olefsky (2021) Insulin resistance is caused by abnormal secretion of insulin from beta-cells of the pancreas. Due to obesity, inactive lifestyle and some genetic predispositions beta-cells experience stress and exhibit a continuous decrease in their functioning and decline in the capacity of beta-cells to secrete insulin, thus it caused them to secrete abnormal concentrations of insulin (Courtney & Olefsky, 2021). Courtney and Olefsky further added that mostly this decline in insulin secretion is observed in the liver and muscle as the liver and muscle are mostly involved in the absorption of carbohydrate through lots of channels in their cell membrane, but the decline in insulin concentration can also occur in the gastrointestinal tract, adipose tissue, kidney, and the brain. The metabolic consequences of abnormal insulin concentration in different cells of the body lead to hyperglycemia, hyperuricemia, and hypertension (Courtney & Olefsky, 2021).

7.1.4.       Mitochondrial Disfunction:

It is still the topic of debate that whether mitochondrial dysfunction is the cause of insulin resistance or the result of insulin resistance. According to Courtney and her colleague, impaired secretion of adiponectin and an increase in the number of reactive oxygen species is associated with mitochondrial dysfunction in adipose tissue and non-adipose tissue respectively (Courtney & Olefsky, 2021).

7.2.  Alpha-Glucosidase

The alpha-glucosidase enzyme is a hydrolase that converts starch of dietary sources into simple sugars like glucose to fulfill the glucose requirements of the body. Humans require this enzyme to use glucose from complex carbohydrate sources. Dietary sources do not contain glucose in the simple form and the complex form cannot be absorbed through the small intestine.  In the absence of deficiency of these alpha-glucosidase in a normal person blood plasma can run out of glucose, loss of ATP can occur and it can lead to serious complications and death. But, According to Hedrington and Davis (2019) in their publication about the “Considerations while using alpha-glucosidase inhibitors in the treatment of Type 2 DM”, hyperglycemia (increase in blood glucose concentration more than normal) is the most common complication in Type 2 DM and alpha-glucosidase is the source of increase in blood glucose, inhibition of alpha-glucosidase can be the treatment for the patients with Type 2 DM (Hedrington & Davis, 2019).

Moreover, according to Hedrington and Davis (2019), alpha-glucosidase inhibitors are the class of non-invasive drugs that can cause mild side effects including diarrhea. But, their side effects are dose-dependent and short-lived. Thus, the side effects are cured within a short time and can be managed by reducing the dose of the drug (Hedrington & Davis, 2019). Sometimes, the complications in diabetes are caused by the poor dietary habits of the patient. Doctors always recommend that diabetic patients avoid the consumption of exogenous sugar but not following these guidelines of the doctor's patients often cannot restrict themselves from sugar consumption leading to the consequences of hyperglycemia. Alpha-glycosidase inhibitors could be the savior in such situations and these drugs can be prescribed when the patient has intentionally or non-intentionally consumed a large amount of drug. Before letting this glucose reach the blood and cause type 2 DM complications the glucose can be made non-available to the blood by restricting its absorption from the intestine.

According to Dirir et al (2021) until now only three alpha-glycosidase inhibitors are being utilized in clinical trials which are given as acarbose, miglitol, and voglibose (Dirir et al., 2021b) Their structures are given in Fig 1.1. Therefore, it is required to find more inhibitors for alpha-glycosidase with increased efficiency.

Fig 1.1

Text Box: Voglibosee                                                                      

Text Box: Miglitol

 

                                   

Text Box: Acarbose

                                                                                                                         

7.3.  In-Silico Drug Designing

            In- Silico is a Computer-aided method for drug design playing an ever-increasing role in drug discovery that is important in the cost-effective recognition of promising drug candidates.

According to Chandrasekaran et al (2018) in-silico, drug designing is extensively applied in drug discovery and designing. In this process, recognition of the suitable drug target is the first and foremost task. Recognition of such targets is necessary to expose a sufficient level of ‘confidence’ and to know their pharmaceutical application to the disease under inspection (Chandrasekaran et al., 2018)

According to Guan et al (2019) in silico drug designing is the need of the time. One of the benefits of this technique is that it requires less time, less financial resources, and is more effective. According to various needs of the experiments and differentiating nature of compounds, in-silico drug designing or computational drug designing is classified into two groups, which are structural-based drug designing and ligand-based drug designing (Guan et al., 2019). Ligand-based drug design will be used in this research.

7.4.  Ligand-based approaches in drug repurposing:

Drug repurposing is the advancing field of drug designing in which the previously approved drugs are studied to check if they have an affinity for other compounds (unintended targets) and if they can be used to treat another disease. Drug repurposing is also named drug repositioning. Thus by repurposing drugs scientists design new treatments from the same drug and these newly designed drugs are not needed to be tested in clinical trials as they have previously been approved and have gone through clinical trials (sciences & 2013, n.d.).

            According to Gaulton et al (2017), in drug repurposing, the ligand-based approach is the method of repurposing a drug by assuming that ligand with similar structure will have similar properties and thus can be utilized in similar functions (Gaulton et al., n.d.). Databases like drug bank, ChEMBL, and PubChem contains thousands of compounds that are from various sources. Some of these compounds are approved as drugs, some are reported for various biochemical activities. Researchers in the ligand-based approach of drug repurposing take compounds to form these databases and use them to check if they have interactive properties for any of their targets.

Accordingly, ligands are small compounds that have been isolated in the laboratory by scientists and their structures have been determined through various techniques like X-ray crystallography, and after that their structures have been submitted in the in silico databases. Thus, these structures make a library of compounds in the form of databases.  These databases are the representation of the huge reservoir of different kinds of information like their ADMET properties (Properties about the Absorption, Distribution, Metabolism, Excretion, and Toxicity), binding affinity, and their different structural aspects (Gaulton et al., n.d.). According to Shameer et al (2017), the advances in in-silico drug repurposing have developed databases that contain properties like bioactivity data and therapeutic properties (Shameer et al., n.d.).

According to Gaulton et al (2017), the databases (e.g. ChEMBL, drug bank, and PubChem) have 3D structures of the compounds these compounds can be downloaded in different forms and they can be prepared using different software tools as if they are being prepared in the laboratory. After that, they are interacted using different online or offline tools and their interactions can be seen with the target compound. Good interactions in the in silico experiments are the evidence of their good supposed interactions in vitro and in vivo (Gaulton et al., n.d.).

7.5.  Molecular Docking

            According to Kitchen et al (2004) molecular docking is a tool for drug repositioning. Several drug designing and drug repurposing approaches can be utilized by using the tool of molecular docking. Whether it is a ligand-based drug designing approach or any other approach the basic principle in molecular docking is the same. Molecular docking tools predict whether two compounds can bind to each other or not. The results are shown as binding affinity and binding energies. The lower the binding energy, the better the drug can bind to the target. On the other hand, the higher the binding affinity, the better the drug can bind to the target (Kitchen et al., n.d.).

Recently it was reported by Dakshanamurthy et al (2012) molecular docking has become very successful in in-silico drug repurposing and drug designing (Dakshanamurthy et al., 2012). As discussed by Kitchen et al (2012) in molecular docking for example if the ligand-based approach is applied) different small compounds (ligands) have been taken from different databases and they are prepared by using different tools after preparation these small compounds are docked in docking software such as auto dock vina. Further stated by Kitchen et al (2012)The interactions of the ligand and the target can be seen by docking and by different algorithms several properties can be predicted. The interactions and other properties predict whether a ligand can be used as an inhibitor for a certain compound or not (Kitchen et al., n.d.).

7.6.  Auto Dock Vina

According to Muzio Auto dock, vina is a molecular docking tool. That is used to check the interactions between any two compounds. One of these compounds is the predicted drug which is chosen due to several previous publications indicating the role of the compound in a certain disease or the property of the compound that predict its affinity for a second compound. While the second compound is a biochemical target that is involved in the prognosis of a disease and its inhibition can cure a disease. The biochemical target is a large compound mostly an enzyme while the ligand is a small compound. The quality of interactions is determined by several inbuilt algorithms in auto dock vina (Muzio et al., n.d.).

7.7.  Phytomedicines:

Phytomedicines are plant-based drugs that are extracted from the roots, shoots, leaves, bark, or any part of the plant. They are plant secondary metabolites and have several different functional groups that have the affinity for binding with the biochemical target molecule in the body in physiological conditions. Till 2021, many plant-based medicines have been approved by FDA and are being utilized for the treatment of different diseases and disorders (Ben Hlel et al., 2021). According to Wu and Charles (2020), these drugs are called botanical drugs and are named Veregen (approved in 2006) and Fulyzaq (also named Mytesi) in 2012. Although it is still a scientific challenge to describe their regulatory consistency (Wu et al., 2020).

Ben Hlel in 2021 argued that For complicated metabolic disorders like diabetes and obesity plant-based drugs can prove to be effective. Going through the history of metabolic disorders, one can find that whenever conventional medicines have no way out and no cure for the disorders, traditional medicines always have shown their potential as stated by Ben Hlel (Ben Hlel et al., 2021).

8.            The Research Model

Figure 1.2

 

 

 

 

 

 

 

 

Figure 1.2: Research Model

In this research model, some simple but expertized steps will be performed and different software will be utilized at different steps. As indicated by the model the first step will be the Selection of both the ligands and the target. The second step will be the preparation of both the ligands and the target. In the third step both the prepared ligands and the target will be interacted by using molecular docking software. Auto dock vina will be used as molecular docking software in this research. In the end, the results will be interpreted based on different values shown by the autodock vina. These values will indicate the strength of the interaction of the ligands and targets and thus predict the possibility of the binding of the ligand and the target. Precisely speaking the ligands will be the terpenoids extracted from the plant and the target molecule will be the alpha-glucosidase.

Furthermore, the molecular docking strategy that will be used in this study is ligand-based drug designing and the research analysis is quantitative.

9. The Hypothesis of the Research Study

Plant terpenoids will prove to be the potential inhibitors for the enzyme alpha-glucosidase contributing to the treatment of diabetes mellitus type 2.

10. Research Gap

Till now there is no conclusive treatment for Type 2 DM that can curatively reduce the metabolic dysfunction and complications related to Type 2 DM. According to Aziz (2012), the most considerable cause of the complications of Type 2 DM is hyperglycemia. Various approaches have been utilized to treat the complications of Type 2 DM by treating hyperglycemia, which is the increase in plasma or blood glucose concentration more than normal. Further, Aziz stated that among these approaches and treatments most famous ones are the use of injectable insulin for the treatment of Type 1 DM and the use of drugs in combination with injectable insulin in the case of Type 2 DM (M. A. Aziz, 2012). But, Chatterjee and Davies (2015) in “Current management of Diabetes Mellitus and future directions” added to this that rather than treatming Type 2 DM by injecting insulin that could be harmful and can contribute to insulin resistance and other physiological complications Type 2 DM can be treated by using medications than will directly target the factors that are increasing the glucose concentration in the plasma contributing to hyperglycemia. These medications are termed as non-insulin medicines and are more useful for the healthy life style of the diabetic patients (Chatterjee & Davies, 2015).

As investigated by Bhowmick and Banu in their recent publication about the “Therapeutic targets of Type 2 Diabetes ” in 2017, a way to manage the glucose concentration in the plasma is to suppress the absorption of glucose from the intestine so if it is being ingested through the exogenous way it will not reach to the blood it can be done by inhibiting the enzymes that are responsible for the absorption of glucose from the intestine. Another way is to suppress gluconeogenesis (synthesis of glucose from non-carbohydrate sources) from the liver. This gluconeogenesis is called hepatic gluconeogenesis (Bhowmick et al., n.d.). Further added by Bhowmick and Banu (2017) a third dominant way to maintain glucose concentration in the plasma or blood is to decrease reabsorption of glucose through kidneys. Glucose, when filtered through kidneys, is often taken back in the blood from the kidneys this is called the reabsorption of glucose from the kidneys. All of these three ways of treating hyperglycemia are termed non-insulin medications and the most prominent non-insulin medications include alpha-glucosidase inhibitors and sodium-glucose co-transporter 2 inhibitors (Bhowmick et al., n.d.).

Recently many researchers have proved the importance of alpha-glucosidase as a target enzyme and many classes of compounds have been docked and virtually screened against it. Hedrington and Devis in 2019 and Dirir and his colleagues in 2021, argued that the inhibition of Alpha-Glucosidase is explicitly involved in treating diabetes mellitus type 2. Alpha-glucosidase should be studied at a higher level to evaluate and deploy its capability for the treatment of one of the most frequent metabolic disorders of the world; diabetes mellitus type 2 (Hedrington & Davis, 2019) (Dirir et al., 2021b). The most common anti-diabetic drugs that work by inhibiting alpha-glucosidase and retarding carbohydrate digestion include Acarbose- Precose or Glucobay (Hedrington & Davis, 2019) (Dirir et al., 2021b).

            Further Dirir (2021) added that various medicinal plant extracts have been studied for suppressing alpha-glucosidase and they have shown surpassing outcomes than acarbose but they interfere with gut flora and produce anti-nutritional effects. After acquiring satisfying results, selected compounds were passed through different phases of clinical trials before reaching the market. Previously endorsed drugs should be screened against alpha-glucosidase to identify if any of them can be repurposed as a potential inhibitor of alpha-glucosidase (Dirir et al., 2021b).

11. METHODOLOGY

11.1 Research Design

The research design is ligand-based drug designing. Plant terpenoids are selected as ligands as some terpenoids have shown good interactions with alpha-glucosidase. Based on the ligand-based drug design approach the related compounds will suppose to be showing good interactions with the alpha-glucosidase.

Furthermore, the research model shown in figure 1.2 will be followed. The first step, the selection of ligand and the target molecule has been performed. As discussed earlier the plant terpenoids are selected as the ligands and alpha-glucosidase enzyme is the target molecule. The second step will be performed now which is the preparation of the active sites of both ligands and target molecules. Then they will be docked in auto dock vina software.

11.2 Sampling Procedure and Sample Size

Following the in-silico sampling methods, the procedure of sampling will be the downloading of required compounds from the relevant databases and their libraries will be made. Almost, 200 plant terpenoids will be selected as ligands and collected in the form of a virtual library.

11.3 Sampling Technique

The drug repurposing technique is utilized in this study. In this sampling technique plant terpenoids will be screened against a targeted enzyme to repurpose and reposition their role in diabetes mellitus type 2 (T2DM).

11.4 Tools of Data Collection

Following databases will be used as the tools of data collection.

·         PubChem

·         Protein Data Bank

11.5 Data Collection Procedure and Methods

For the collection of the data, PubChem will be used as a protein database. PubChem has thousands of small molecules reported by researchers and the 3D structures of these small molecules are available that can be downloaded and collected in one place. Plant terpenoids will be found from PubChem by searching their names one by one and by downloading their SDF file one by one. If the SDF file of any compound will not be available. Its structure can be manually drawn using drawing software like the Chemdraw. Or the canonical smile or isomeric smiles of these ligands can be pasted in this software.

Moreover, the 3D structure of the alpha-glucosidase will be downloaded from the Protein Data Bank in the PDB file formate.

11.6 Data Analysis Methods and Software

All the collected data will be analyzed by using the following two software.

·         Open Babble

·         Auto Dock Vina

Open Babble is a simple software that can convert ligand files from one format to another format. Any input format can be converted to any open babble supported output format. Open Babble will be used to convert the PDB file downloaded from the Protein Data Bank into a pdbqt file. The ligand files that will be downloaded in SDF format will also be converted to pdbqt files by using Open Babble software. The pdbqt format is the only format supported by auto dock vina.

Moreover, Auto dock vina will be used to perform rapid screening of protein-ligand interactions. After the conversion of enzyme in pdbqt formate by open babble software, the domains of the enzyme will be selected in the form of a grid in Auto Dock vina. It is necessary to select the grid box otherwise the docking will not be performed. A grid box will limit the sites of interactions of the ligands to the targeted enzyme. If there will be no limit in the interactions thousands of possibilities will be found by the software and the processor of the computer will not be capable of performing such a heavy command. Thus the shorter the grid box the easier the will be docking process and lesser time will be required accordingly. Ligands will be provided the grid area to confirm their position of attachment.

Preparation of the grid area will be a crucial process and it will be the process that will provide the direction to the research. The grid area will be prepared by selecting the active domains of the enzyme. Active domains will be found by reviewing the literature. The literature will be studied that accordingly. Active domains will be found by finding the sites where the ligands were previously bounded when the enzyme will be downloaded in PDB format. In case of the presence of more than one domain, more than one grid boxes will be made and docking will be performed more than one time and the results of all the grid boxes will be interpreted separately. This process will make the research more precise and effective. The topmost effective ligands will be selected based on the lowest binding energies of the ligands.

 11.7 Expected Results

As some terpenoids have shown good binding with the alpha-glucosidase it is expected that by following the ligand-based drug designing approach the terpenoids that will be used in this research will show good binding as well. Although all of the ligands may not have an effective docking score, it is expected that most of the ligands will be found to be potent inhibitors of alpha-glucosidase.

11.8 Time Frame to complete the Research Study

1.      Data collection will require about a month

2.      Data preparation will require a month as well

3.      Docking will take about a week

4.      Manuscript writing will take about one month

12. Total Required Time:

As it is the computational drug designing, it will require a lesser amount of time as compared to the in vitro or in vivo approaches. With the prowess and proficiency in all the tools mentioned in the research proposal, this whole research will be completed within 4 months with the completion of the manuscript as well.

13. Proposed budget:

There will be no need to buy any samples, particularly for this study. The research will be performed with the help of a good quality laptop and no budget will be required.

14. Required apparatus:

This study will require a laptop with a good working processer. Furthermore, some software will be required as described previously in the proposal. This software will include Docking software like Auto Dock vina. Moreover, Open Babble will be required for forming pdbqt files and Pymol will be required to view the interaction between ligands. Also, Chemdraw software will be required to draw compounds that will not be downloaded from the PubChem in the form of SDF files.

15. Discussion

Type 2 diabetes is the disability of the body to regulate and use sugar (glucose) as a fuel which causes a large amount of sugar to circulate in the bloodstream. This high sugar level eventually leads to other disorders of the circulatory, immune, and defense systems. Furthermore, type 2 DM can cause various microvascular and macrovascular complications.

There are two main reasons for this poor regulation of body sugar. One is that the pancreas does not release enough insulin and the other is that the cells badly respond to insulin and take in less sugar due to which more glucose circulates in the blood. To compensate for the bad eating habits of the patients it is necessary to develop medicines that can stop the increase in blood glucose concentrations even if a high glucose diet is taken by the patient. It can be done by stopping the conversion of glucose from the dietary complex carbohydrates and eventually inhibiting the absorption of the glucose through the small intestine in the blood.

Alpha-glucosidase is a hydrolyzing enzyme that breaks down the starch of the food in the form of glucose to make the carbohydrate available to the blood. Thus, the food items comprising of complex carbohydrates are metabolized by alpha-glucosidase in the pancreas and transformed into glucose. This glucose is fatal for T2DM patients. Inhibiting alpha-glucosidase diminishes the formation of glucose by hydrolysis thus exogenous glucose becomes no longer available to damage the body.

Plant-based medicines have been proved to be multipurpose and beneficial especially in the case of metabolic disorders like diabetes and obesity. Phytomedicines are the traditional medicines that have long been used by ancient people but the biochemistry behind them was unknown till the advancements in the sciences. Nowadays various plant extracts are being tested to check their affinities for different targets. Several plant-based medicines have been approved by FDA as well as discussed in the proposal earlier.

Moreover, there are only three small molecules (Acarbose, Miglitol, and Voglibose) are in clinical trials. Thus there is a need to test more plant-based compounds again the alpha-glucosidase to find more effective and potent inhibitors. For this purpose, various plant terpenoids are extracted from different sources which is the main focus of this research. By analyzing the binding affinities of these Ligands with the targeted enzyme many other potent drugs can be identified which can play major role in the treatment of type 2 diabetes mellitus.


 

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