Pharmacological Inhibition of Mutant ALK for the Treatment of Non-Small

Cell Lung Cancer:

Muneeb Rasool1, Muhammad Sajjid2

University of Okara*all , Renala Khurd, Okara

Abstract:
Lung cancer is the most common among all the types of cancer worldwide with1.8 million
people diagnosed every year, leading to 1.6 million deaths every year according to the American
cancer society. The involvement of mutated Anaplasic Lymphoma Kinase (ALK) positive fusion
protein in the progression of NSCLC has made a propitious target to inhibit and treat NSCLC. In
the present study, the main motif is to screen the most effective inhibitor against ALK protein
7LIR with the potential pharmacological profile. The ligands selected were docked with
Molegro Virtual Docker (MVD) andCEP-37440 (PubChem CID- 71721648) was the best
docked pre-established compound with a permissible pharmacological profile.

Introduction:
Lung cancer remains the major cause of mortalities related to cancer. In the year 2016, there
were 224,390 new lung cancer cases diagnosed and 158,080 deaths occurred in the US. It is
more common in males than in females. (Gascoyne, Aoun et al. 1999)It retains the second
leading cause of deaths in females following breast cancer.NSCLCconstitutes 70-80% patients of
lung cancer and is further categorized on the basis of the site of its occurrence adenocarcinoma
(outer part of the lungs), squamous cell carcinoma However, in diseased state, it undergoes a
translocation mutation which leads to the initiation of cancer.(Roskoski Jr 2013) The molecular
mechanism of this translocation mutation involves the inversion of the 2nd chromosome, with
subsequent joining of exons1-13 of EML4 to exons 20-29 of alk, which results in the synthesis
of the abnormal chimeric protein functioning as tyrosine kinase.(Sharda, Khandelwal et al. 2019)
The different isoforms ofALK-EML4 fusion product have been reported (as a result of different
combinations brae– fusion point between the two exon sets) and various studies have shown the
involvement of all these fusion proteins in cancer development The chimeric protein also forms
chimeric fusions with other proteins like nucleophosmin. (Minguet, Smith et al. 2016)Several
cell lines as well as mouse models have shown the involvement of the chimeric protein in
tumorigenesis Transgenic mouse having ALK chimeric protein showed the onset of carcinogenic
nodules throughout the lungsearly after the birth and this pathological condition was reversed
with ALK -specific inhibitor.(Duyster, Bai et al. 2001)
Lung cancer is a type of cancer that starts in the lungs. Cancer starts when cells in the body begin
to grow out of control. iYour lungs are 2 sponge-like organs in your chest. Your right lung has 3
sections, called lobes. Your left lung has 2 lobes. The left lung is smaller because the heart takes
up more room on that side of the body.(Delsol, Lamant et al. 1997) Lung cancer is a type of
cancer that starts in the lungs. Cancer starts when cells in the body begin to grow out of
control. Your lungs are 2 sponge-like organs in your chest. Your right lung has 3 sections,
called lobes. Your left lung has 2 lobes. The left lung is smaller because the heart takes up more
room on that side of the body.(Mattsson, Turner et al. 2010)

Non-small cell lung cancer :

About 80% to 85% of lung cancers are NSCLC. The main subtypes of NSCLC are
adenocarcinoma, squamous cell carcinoma, and large cell carcinoma.(Minguet, Smith et al.
2016) These subtypes, which start from different types of lung cells, are grouped together as
NSCLC because their treatment and prognoses (outlook) are often similar. Squamous cell
carcinomas start in squamous cells, which are flat cells that line the inside of the airways in the
lungs. They are often linked to a history of smoking and tend to be found in the central part of
the lungs, near a main airway (bronchus).(Dardaei, Wang et al. 2018)

Materials and Method:

Selection of Ligand:

Bromide, Alectinib, Riboflavin , Ceritinib ,Brigatinib , ASP3026 , Oxalate ,

Luteolin ,Carbonate , Acetate.

Selection of Protein:

First of all, select the protein (7LIR) from PDB web site and download in the PDB format. 7LIR
protein run on chimera soft wear remove the chain except A chain, remove the ligand and water.
Minimize the protein and save in PDB.

Zinc database: The structures of ligand are obtained from ZINC database and download in SDF
Format. Then these SDF Format ligands are open in Chimera one by one and download their
structure in PDB format.

Chimera: To minimization of gene/protein structure, the tool chimera is used. Before

minimization of structures, chains, residues, ligands are removed

Docking:
Pyrx: Used to see the binding affinity of ligand and protein in complex.

Prediction of toxicity:

Protox tool: Used to see the toxicity of ligands.

Docking Results:

Compound Energy Requird

Hesperidin -8.4
Quercetin -6.8
Bromide -4.8
ceritnib -6.0
Brigatinib -5.9
ASP3026 -6.6
Proralen -7.5
Acetate -2.8
luteolin -6.0
Carbonate -2.9
Table No. 1: This table show the binding affinity of compound/ligand histoprotein show
maximum energy and carbonate show minimum binding energy.

Discussion:
Our study examined the pharmacological effects of cortex more medicinal ingredients in lung
cancer. The protein 7LIR is most effective protein in thiseliginds.disease targets via network
analysis. Hesperidin, .as the active ingredients related to the most targets. The drug-likeness
value of brigitinib. among the major active ingredients in cortex more. In addition, docking
studies showed that citinib.has a good affinity for the target protein 7lir. In nature, cyclomorusin
is a common flavonoid compound. Many studies have shown that most of the flavonoids
compound in traditional Chinesemedicine have strong antioxidant activity .Cyclomorusin has
been confirmed to have the dual effect of inhibiting and promoting uric acid excretionUntil
present, there is no relevant literature on cyclomorusin in lung cancer cells. iiTherefore, this
experiment provides a good reference value for the future study of cyclomorusin in moretumor
cells. Traditional Chinese medicine has the characteristic of less side effects, which provides a
new treatment strategy for improving the side effects of chemotherapy drugs in clinical practice
However, the specific ingredients of traditiona.iiiCell biology experiments in vitro showed that
cyclomorusin, the active component of cortex mori, had acertain inhibitory effect on lung cancer
cells in a time- and dose- concentration dependent manner andhad additive effect with cisplatin
against lung cancer cells. When treated lung cancer cells withcyclomorusin or cisplatin for 24h,
the results of flow cytometry analysis revealed that cyclomorusin promoted the apoptosis of lung
cancer cells significantly better than cisplatin. Relevant studies have confirmed that inhibition or
reduction of phosphorylation of PI3K/AKT signaling pathway related proteins can inhibit the
occurrence and development of tumor cells. A number of studies have demonstrated abnormal
activation of PI3K/AKT signaling in malignant tumors, which may have a bearing on the
occurrence and development of tumors, regulates cell proliferation, survival, transformation,
adhesion and extracellular matrix degradation, and participates in the whole process of liver
cancer.

CONCLUSION.
I have concluded that this ligands Hesperidin if we inhibit their function in lung than no cause
the lung cancer in human. A ligand HESPERDINE -8.4 which value is highest it is very useful
for inhibition. In addition, the pharmacological effects of cortex more and the major ingredients
cyclomorusin were comprehensive discussed from both theoretical and practical aspects by
network pharmacological analysis combined.

References:
Dardaei, L., et al. (2018). "SHP2 inhibition restores sensitivity in ALK-rearranged non-small-cell
lung cancer resistant to ALK inhibitors." Nature medicine 24(4): 512-517.

Delsol, G., et al. (1997). "A new subtype of large B-cell lymphoma expressing the ALK kinase
and lacking the 2; 5 translocation." Blood, The Journal of the American Society of Hematology
89(5): 1483-1490.

Duyster, J., et al. (2001). "Translocations involving anaplastic lymphoma kinase (ALK)."
Oncogene 20(40): 5623-5637.

Gascoyne, R. D., et al. (1999). "Prognostic significance of anaplastic lymphoma kinase (ALK)
protein expression in adults with anaplastic large cell lymphoma." Blood, The Journal of the
American Society of Hematology 93(11): 3913-3921.

Mattsson, G., et al. (2010). "Can cytoplasmic nucleophosmin be detected by

immunocytochemical staining of cell smears in acute myeloid leukemia?" Haematologica 95(4):
670.
Minguet, J., et al. (2016). "Targeted therapies for treatment of non‐small cell lung cancer—
recent advances and future perspectives." International journal of cancer 138(11): 2549-2561.

Roskoski Jr, R. (2013). "Anaplastic lymphoma kinase (ALK): structure, oncogenic activation,
and pharmacological inhibition." Pharmacological research 68(1): 68-94.

Sharda, S., et al. (2019). "A computer-aided drug designing for pharmacological inhibition of
mutant ALK for the treatment of non-small cell lung cancer." Current topics in medicinal
chemistry 19(13): 1129-1144.
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