CHAPTER 4

RESULTS

4.1 SELECTION OF RECEPTORS AND PROTEINS

4.1 Selection of receptor and proteins
The receptors co-crystallized with ligands were obtained from the Protein Data Bank(PDB). The selection of the
receptor was based on the understanding of the effects of either its upregulation or downregulation."Figure
4.1a, 4.1b, and 4.1c represents the PDB 3D format of 4NKW and 2YHD respectively;

Figure 4.1a : 3D structural representation of 4nkw

Figure 4.1b : 3D structural representation of 2yhd
4.2 Validation of docking protocol
The validation of the docking protocol aimed to ensure in-silico reproducibility or close reproducibility of the
experimental protein-ligand interactions obtained from the Protein Data Bank for the protein. The results of the
docking validations are shown in the figure below;

Figure 4.2a :super imposed view of 4NKW reference compound in green and docked reference compound
in blue.
Figure 4.2b: super imposed view of 2yhd REFRENCE compound in green and docked refrence
compound in blue .
4.3 Molecular docking of the phytochemicals
The molecular docking of the phytochemicals was conducted on proteins 4NKW and 2YHDto assess the binding
activity between the phytochemichals and the proteins active site.Phytochemicals exhibiting superior binding
affinities compared to reference standards and existing drugs were identified by their lower binding energies. The
mean binding energies were calculated and presented in the table below. These promising phytochemicals were
characterized by lower binding energy values as lower
binding energy indicates stronger
ligand binding, with the significance of binding energy values determined
by the largest negative magnitude, indicating the most favorable
conformation of the ligand-protein complex formed when the ligand
effectively binds to the protein's active site. The table also presents the standard deviation of
the phytochemicals with better binding affinities than that of the reference compounds and already existing drugs
The table below represents the standard deviation and mean of the phytochemicals with better binding affinities
than that of the reference compounds and already existing drugs,
Table 4.1a; Phytochemicals with better binding energy values against 4NKW with their mean and standard
deviation.

4.4 post docking analysis

25 phytochemicals of higher affinities with multi targeting functions against the two proteins obtained
in comparison with reference compounds and used for further studies are shown in the table 4.2 below;
S/N PHYTOCHEMICALS
1 ursolic acid
2 vincristine
3 vinblastine
4 ergosterol peroxide
5 dehydroergosterol
6 Rg5
7 oleanolic acid
8 ocotillos
9 bufadienolide
10 artemisinin
11 indicaxabthin
12 curcumin
13 proanthrocyanidins
14 aloesin
15 lapachol
16 delta -9
17 kigelinone
18 daucosterol
19 kaempferol
20 aloresinA
21 chlorogenic acid
22 B- sitosterol
23 protopanaxatriol
24 protopanaxatriol
25 lycophene

4.5 Assessment of protein-ligand interaction

The protein-ligand interaction of the front runner phytochemicals was assessed using discovery studio
visualizer and it revealed that for protein 4NKW, the percentage of amino acids that interacted with the reference
compound is within the range of

4.6 Swiss target prediction

The results obtained from the swiss target prediction showed that the protein class targeted by the reference
componds is also targeted and binded by the forerunner phytochemicals as-shown below;

Table 4.4; Front runner Phytochemicals for 4NKW and 2YHD with the
class of protein targeted or binding to;

4.7 Phytochemical analysis.

Results of phytochemical screening of the extracts of Melissa officinalis, moringa oleifera, Catharanthus roseus are
shown in table 4.5 below

S/N Melissa moringa Catharanthus

officinalis oleifera roseus
1 Meyer’s test for + - +
alkaloids

2 Hager’s for + - +
alkaloids

3 Dragendoff’s test + - +
for alkaloids

4 Lead acetate test + + +

 for flavonoids

5 Sodium hydroxide - + +
test for flavonoids

6 Saponins - - -
7 Tannins + + +
8 Steroids + - +
9 Phlembotannins + - -
10 Terpenoids + + +
11 Cardiac glycosides + + +

Key
+ for positive
- for negative