Introduction

There are several data which indicate that cancer has increased in recent years worldwide,^{[1, 2]} causing a decrease in life expectancy population.^{[3, 4]} Several risk factors have been observed for cancer development such as alcoholism,^[5] obesity,^{[6, 7]} cigarette smoking,^[8] and foods rich in fat.^{[9, 10]} Furthermore, a study indicates that different genetic factors may be involved in different types of cancer cells;^[11-16] for example, a report displayed that mutations in the KRAS gene (Kirsten rat sarcoma virus) are related to cancer cell growth.^[17] Another study conducted in 436 French people with cancer indicates that 348 patients showed an established deleterious mutation in MSH2 (tumor suppressor gene).^[18] Besides, a report suggests that BCR-ABL gene mutations (which encode for a tyrosine kinase) are linked to leukemia developed.^[19] Another study indicates that mutations involved in HER2 (a membrane tyrosine kinase) are associated with breast cancer.^{[20, 21]} Furthermore, a study indicates that overexpression of cMYC (regulator gene that codes for transcription factors) is associated with breast cancer.^[22] Another report indicates that mutations in the EGFR gene are related to lung cancer cell growth from 15 to 20% which 10% correspond to kinase domain insertions in exon 20.^[23]

Materials and Methods

Figure 1 depicts the structure of twenty-six carbazole derivatives, which were utilized to ascertain if they may interact in the following ways with the JAK3 surface:

Ligand-protein complex

Interaction of twenty-six carbazole derivatives with JAK3 was evaluated using 3pjc protein,^[32] decernotinib and facitinib as controls in DockingServer program.^[33]

Pharmacokinetic parameters

Different pharmacokinetic factors for carbazole analogs (2, 5, 9, 17, 18, and 22) were determined with the use of the SwissADME software.^[34]

Toxicity analysis

The GUSAR software was utilized to assess the potential toxicity of carbazole analogs (2, 5, 9, 17, 18, and 22) as well as decernotinib and facitinib when administered through various routes.^[35]

Results and Discussion

Certain observations in the literature suggest that analogs of carbazoles may cause changes in the development of cancer cells; for example, alectinib drug has been used as monotherapy to treat lung cancer.^[36] Another study showed that compound 3,6-di(2-furyl)-9H-carbazole can act as a carcinogenic agent through topoisomerase II inhibition using MCF-7 (breast cancer) cell lines.^[37] Furthermore, a report displayed that some N-acylated carbazole derivatives can decrease breast cancer cell growth using MDA-MB-231 cell lines.^[38] Another report showed that a tetrahydrocarbazole derivative exerts anticancer activity against lung cancer using a Calu-1 cell line.^[39] Besides, a study indicates that the compound MHY407 can produce biological activity in breast cancer using an MCF-7 cell line.^[40] All of these studies suggest that some drugs are used to control the growth of cancer cells; however, their interaction with JAK3 in cancer cell growth is unclear. Thus, this study aimed to characterize the interaction of twenty-six carbazole analogs with JAK3 using 3pjc protein, decrnotinib, and facitinib as theoretical tools in the DockingServer program.

Protein-ligand

There are several protocols to characterize the protein-ligand complex formation; for example, a study displays the interaction of a carbazole-acetate derivative with tyrosinase and human glutathione receptors using the Autodock program.^[41] Other studies suggest that some carbazole analogs can interact with peroxisome proliferator-activated receptor gamma using GROMAC software.^[42] Besides, a report indicates that a carbazole-chrome-carboxamide analog can interact with cyclin-dependent kinase 2 using the Biovia Discovery Studio program.^[43] These theoretical data suggest that some carbazoles interact with different biomolecules; therefore, the objective of this research was to characterize the coupling of twenty-six carbazoles with JAK3 using DockingServer software.^[33] Table 1 and Figure 2 displayed the possible interaction of carbazole derivatives with 3pjc protein through coupling with different amino acid residues. In this way, the results indicate that Leu₈₂₈ could interact with carbazole analogs 1, 3-21, and 24 in a similar manner to decrnotinib and facitinib; these data suggest that possible biological activity produced by carbazole analogs could be conditioned by the interaction with different types of amino acids involved in carbazole-JAK3 complex formation. However, it is crucial to note that the formation of carbazole-protein complexes may be influenced by certain thermodynamic parameters.

Thermodynamic parameters

In the literature, there are some protocols to predict Protein-ligand complex formation.^[33] This research used the DockingServer program to characterize some thermodynamic parameters that can influence the formation of the carbazole-protein complex. Table 2 shows variations in energy levels for carbazole analogs compared to both decernotinib and facitinib drugs. Furthermore, the inhibition constant (Ki) was lower for carbazole analogs 2, 5, 9, 17, 18, and 22 compared with either decernotinib or facitinib drugs. These data suggest that carbazole derivatives 2, 5, 9, 17, 18, and 22 may be good JAK3 inhibitor agents. However, to confirm these findings, more research needs to be carried out using some biological models.

A = Est: Free Energy of Binding (kcal/mol); B = Est. Inhibition Constant, Ki (mM)

C = vdW + Hbond + desolv Energy (kcal/mol); D = Electrostatic Energy (kcal/mol)

E = Total Intermolec. Energy (kcal/mol); F = Interact. Surface.

Pharmacokinetic evaluation

Various substances with anticancer action can have their pharmacokinetic parameters predicted and analyzed using a variety of techniques; in this way, a report displayed the pharmacokinetic properties of several carbazoles using some theoretical programs to evaluate the possibility of some of these could have biological activity orally in humans.^[44] For example, the pharmacokinetic properties of compound SR13668 were determined using the system version of the data from software 10.2.^[45] This data suggests that some theoretical programs can be used to predict different pharmacokinetic parameters; in this way, this study aimed to characterize several pharmacokinetic factors for carbazole analogs 2, 5, 9, 17, 18, and 22 using the SwissADME program (Table 3). The determined data showed the possibility of variations in the metabolism of carbazole analogs, involving different types of cytochrome P450 isoenzymes. In this way, compound 2 shows low absorption compared with carbazole analogs 5, 9, 17, 18, and 22. Furthermore, the compound could act as an inhibitor of CYP2C9, which could cause changes in the biological activity of other drugs that are substrates of this isoenzyme such as celecoxib and flurbiprofen (nonsteroidal anti-inflammatory drugs),^[46] glibenclamide and gliclazide (antidiabetic agents).^[47]

Toxicity analysis

Some data suggest that several carbazole analogs may produce toxicity in different biological models;^{[48, 49]} for example, a report showed that carbazol‑oxadiazol‑one analog can produce a higher toxicity degree compared with a carbazol‑acetyl derivative.^[42] Analyzing these data, in this investigation the toxicity degree of carbazole derivatives 2, 5, 9, 17, 18, and 22 was determined using the Gussar program. The results indicate that compound 9 requires a higher dose through the intraperitoneal route compared to carbazole analogs 2, 5, 17, 18, and 22. These data suggest that the level of toxicity is associated with both dose and administration routes.

IP - Intraperitoneal route of administration

IV - Intravenous route of administration

Oral - Oral route of administration

SC - Subcutaneous route of administration

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