Molecular Docking studies of THC-HCA on Cancer Receptors

 

S Narendra Kumar1, Chetan D M2, Lingayya Hiremath3, Ajeet Kumar Srivastava4, Muralidhara P L5, Jyothsana R6, Rithika Pravin Iye7, Ruchika Pravin Iyer8

1,3,4Assistant Professor, Department of Biotechnology, RV College of Engineering, Bangalore.

2Associate Professor, Department of Biotechnology, NMAMIT, NITTE, Karkala.

5Assistant Professor, Department of Chemical Engineering, RV College of Engineering, Bangalore.

6,7,8BE-Graduate Students, R V College of Engineeering, Bengaluru.

*Corresponding Author E-mail: narendraks@rvce.edu.in

 

ABSTRACT:

Cancer is a group of diseases which involves abnormal cell growth that can spread to other parts of the body. There are certain characteristics that are required to produce a malignant tumour. They include:  Avoidance of programmed cell death and uncontrolled cell division to name a few. The progression from normal cells to cells that can form a detectable mass to outright cancer involves multiple steps known as malignant progression. Cannabinoids are among the treatment available. Recent research has shown that cannabinoid coupled with hydroxy citric acid can be used in the treatment of melanoma. Hydroxy citric acid is a derivative of citric acid well known for its anti-obesity and anti-inflammatory property. The current project involves the coupling of tetrahydrocannabinol with hydroxy citric acid to develop a novel drug molecule for potential cancer treatment. Simulation tool like Argus lab is used to construct the 3D structure of the novel drug. Autodock was used to bind the drug with the receptors like TRPV1, CB1, CB2, 4QO1 and 2YGB.

 

KEYWORDS: Hydroxycitric acid (HCA), cannabinoids, receptors, tetrahydrocannabinol (THC), ligand, docking, anti-inflammation, cancer therapeutics, p-53 proteins, Brest Cancer and Neuro cancer.

 

 

INTRODUCTION:

Hydroxycitric acid is a derivative of citric acid well known for its anti-obesity and anti-inflammatory property. The current project involves the coupling of tetrahydrocannabinol with hydroxycitric acid to develop a novel drug molecule for potential cancer treatment. Simulation tool like Argus lab is used to construct the 3D structure of the novel drug.

 

METHODS:

THC-HCA construct was used as the ligand for the studies. This molecule was constructed using the following reaction in Argus Lab tool. Autodock was used to bind the drug with the receptors like TRPV1, CB1, CB2, 4QO1 and 2YGB. The values obtained were noted down in the table Table I.

 

 

The following receptors were chosen for this study:

TRPV1:

The Transient Receptor Potential Vanilloid 1 is a transmembrane protein that belongs to the family of proteins known as

 

 

HCA

THC

THC-HCA complex

Figure1: Molecular Docking in Auto Dock

 

Transient Receptor Potential (TRP) that function as cation channels1,4. TRPV1 gets activated when it binds with capsaicin or when tissue damage is sustained resulting in the production of a sharp pain or a burning sensation5,6. It has been found that the overexpressed TRPV1, during cancer progression undergoes significant downregulation. This can be due to the involvement of TRPV1 in triggering the Fas/CD95 intrinsic and extrinsic apoptotic pathway on sensitization3. Moreover, TRPV1’s involvement in the ubiquitylation of the epidermal growth factor receptor (EGFR) and lysosomal degradation in human epithelial cancer cells by recruiting Cbl via sensitization of the receptor with a suitable ligand4, highlight’s its importance as a cancer drug target. This has been based on studies that have demonstrated THC-acid and THC have antagonistic properties.2

 

4Q01:

The K-Ras gene is known to have some correlation to recovery when treated with cetuximab a monoclonal antibody. Patients having a mutation in this gene show less recovery than those who don’t.7 Mutations in the GTPase-K-Ras have known to create active lesions in humans thus leading to poor response to standard medication, thus active research to find its site of binding is in progress but problems commonly faced will be allosteric regulatory sites.8 K-Ras is an oncogene which is known to regulate homeostasis and tumorigenesis, it is also associated with attenuation of signals in MPAK pathway.9 Thus highlights its importance in cancer drug studies.

 

2 YBG:

There have been studies that indicate p53 is a great suppressor in oncogenic events and eliminates cancer cells through apoptosis.13,10 It also includes DNA damage response, metabolism and stem cell differentiation. p53 activity is regulated through events such as phosphorylation and acetylation.10 It is important to maintain genome integrity and controlled growth. Mutations in p53 are found in the Li–Fraumeni syndrome, a tumor-prone disorder affecting individuals to tumour of the brain, breast, bone and adrenal cortex.11 Most of the patients have mutations in the p53 family or are inactivated.12 Understanding the regulatory pathway will aid in treatment.12

 

CB2 (5ZTY):

The cannabinoid receptor 2 (CB₂) is a G protein-coupled receptor which belongs to the cannabinoid receptor family and in humans it is encoded by CNR2 gene.14  It’s found mainly in the immune system and 2-arachidonoylglycerol (2-AG) is the principle ligand of the receptor and plays a major role in regulation of immune response.15,5 Because of their location and their ability to reduce inflammation, cannabinoids with high binding affinity to CB2 receptors have demonstrated has shown efficiency in the treatment of conditions like Crohn’s disease16,19. There have been studies that show CB2 receptors have the potential to treat conditions that demonstrate hyper-inflammation including Alzheimer’s disease, multiple sclerosis etc. without triggering the psychoactive effects.17,18 This indicates that CB2 receptors may have therapeutic role in the treatment of chronic inflammatory diseases.18

 

CB1 (5TGZ):

Cannabinoid receptor type 1 (CB1) is a G protein-coupled cannabinoid receptor that is encoded by the CNR1 gene in humans.20 It is extensively expressed in peripheral and central nervous system.21. Many studies suggest that the primary function of CB1 is inhibition of neurotransmitter release. The principle ligands of the receptor are endocannabinoids such as anandamide and 2-arachidonoylglycerol (2-AG) and plant phytocannabinoids such as THC which is an active ingredient of the psychoactive drug cannabis.24 Binding of THC to CB1 has been shown to alter immune function in a rodent model which indicates that THC can induce changes in lymphocytes by activating CB1 receptor.22 Also, cannabinoid signalling system in peripheral organs is found to be regulating many pathophysiological activities including cancer.23 It is known that THC downregulates RAS-MAPK/ERK and P13K-AKT cell survival pathways leading to the induction of apoptosis in various types of cancers.25,26,27 Hence the concept of activation of CB1 by cannabinoids may be used in the development of cancer therapeutics.

 

RESULTS AND DISCUSSION:

The following docking results were obtained is shown in the Table I

 

Table I: Docking results

Receptor

Amino Acid

Donor Atom

Binding Energy

Kcal/mol

REF RMS

Root Atom Coordinates

X

Y

Z

TRPV1

(5IRZ)

Serine – 17

Aspartate - 20

Glutamate - 6

Lysine – 13

O

O

O

N

-6.79

-6.90

-6.47

-6.09

2.67

2.53

4.60

4.68

-6.753

-7.211

-7.426

-8.090

3.752

 1.870

-0.890

-1.931

1.063

0.329

0.840

1.261

4Q01

Glutamate-3

Serine-17

O

O

-4.81

- 4.81

12.28

16.45

20.745

-02.835

-17.746

0 0.806

-14.588

00.956

2YGB

Serine -96

Glutamate-171

O

O

-4.75

-4.74

28.30

18.30

6.173

-20.029

-25.943

-11.706

-10.797

- 09.893

CB2 (5ZTY)

Lysine-23

Serine-47

Aspartate-80

Glutamate-50

N

O

O

O

-3.54

-4.84

-5.57

-5.30

48.76

53.10

33.12

51.95

19.567

19.631

5.2

-3.148

-14.182

-12.912

-18.657

07.005

-43.571

-42.629

-45.700

-09.311

CB1(5TGZ)

Aspartate-163

Lysine - 273

Serine – 144

Glutamate- 258

O

N

O

O

-5.84

-5.04

-3.96

-4.72

300.35

303.86

300.46

287.27

49.566

29.382

48.646

42.552

39.858

34.999

38.740

35.629

295.812

278.793

295.276

282.438

 

TRPV1 Receptor

Serine-17

Asparate-20

Figure 2: Molecular Docking Results of THC-HCA on Integral Membrane TRPV1 Receptor at 17th and 20th Amino Acid positions

 

 

Glutamate-6

Lysine-13

Figure 3: Molecular Docking Results of THC-HCA on Integral Membrane TRPV1 Receptor at 6th and 13th Amino Acid Positions

4Q01 Receptor

 

 

Glutamate-3

Serine-17

Figure 4: Molecular Docking Results of THC-HCA on K-Ras Receptor at 6th and 3rd and 17th Amino Acid Positions

2YGB Receptor

 

 

Serine-96

Glutamate-171

Figure 5: Molecular Docking Results of THC-HCA on D13 scaffolding protein Receptor at 96th and 171th Amino Acid Positions

CB2 (5ZTY) Receptor

 

 

Lysine-23

Serine-47

Figure 6 Molecular Docking Results of THC-HCA on Human G Protein Coupled Receptor at 23rd and 47th Amino Acid Positions

 

 

Aspatate-80

Glutamate-50

Figure 7: Molecular Docking Results of THC-HCA on Human G Protein Coupled Receptor at 80th and 50th position of Amino Acids

 CB1 (5TGZ) Receptor

 

 

Lysine-183

Aspartate-193

Figure 8: Molecular Docking Results of THC-HCA on Human Cannabinoid Receptor at 183rd and 193rd Amino Acid Positions

 

 

 

Serine-144

Glutamate-258

Figure 9: Molecular Docking Results of THC-HCA on Human Cannabinoid Receptor at 144th and 258th position of Amino Acids

 

The molecular level docking studies for THC coupled HCA using various cancer receptors were studied. The docking results are as shown in the Table I with its lowest binding energy and positions of amino acids are shown in the Figures 2 to 9. Due to the extensive distribution in neurons the possibility of exploring TRPV1 as a probable drug receptor for tumours in brain and breast are promising and was justifiable with Carlo et al. Mutations in 4Q01 and 2YGB receptors leads to the pancreatic and colon cancers that can be effectively controlled by THC HCA combined analogues was justifiable from the docking results. The results obtained were completely new and nowhere stated in the literature. Finally it can be concluded that the results generated from above molecular simulation studies can be used to design a de novo lead molecule to address the further development in various cancer therapies. Thus THC-HCA analogue apart from its anti-obesity nature serves as a more potent nutraceutical and therapeutic agent for tumour control.

 

CONCLUSION:

The binding energy along with the Ref RMS values have enabled us to conclude that out of the different receptors that were considered, TRPV1 is theorized to be the most effective in binding with the HCA-THC construct. Due to the extensive distribution in neurons the possibility of exploring TRPV1 as a probable drug receptor for tumors in brain is promising. The RMS values allow us to understand the ability of the molecule to reproduce the correct conformation that is the most ideal for binding and hence most suitable. Since, the RMS of the other receptors was very high indicating that meaningful interaction between the receptor and ligand is unlikely.

 

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Received on 31.05.2019           Modified on 07.12.2020

Accepted on 19.08.2021         © RJPT All right reserved

Research J. Pharm. and Tech. 2022; 15(7):3195-3199.

DOI: 10.52711/0974-360X.2022.00535