الفهرس 
Study of the protein binding displacement of co-administered drugs highly bound to serum albumin using different analytical techniques and molecular docking
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Abstract
Study of the protein binding displacement of co-administered drugs highly bound to serum albumin using different analytical techniques and molecular docking
This thesis is concerned with studying the binding interaction of tropisetron with human serum albumin and displacement between Tropisetron and PPIs with human serum albumin by using several spectroscopic techniques , HPLC , voltammetry and molecular docking
SimulationThe thesis consists of the following sections:
Section 1: Introduction It includes:
An overview about plasma protein binding, human serum albumin, different approaches for studying plasma protein binding,displacement drugs and investigated drugs
Section 2: Literature review
Literature review about the method of analysis for tropisetron, and lansoprazole
Section 3: Aim of the work.
IT includes the aim of this work and the basis on which the work has been done
Section 4: Spectroscopic methods of tropisetron and lansoprazole
This section is divided into 2 parts
Part1: Experimental
Part2: Results and Discussion Which includes: UV absorption measurement, Fluorescence spectroscopy measurements, Synchronous fluorescence spectroscopy, FT-IR analysis.
UV absorption measurement part:
The purpose of the study was to compare the spectra of human serum albumin solutions in absence and presence of tropisetron / lansoprazole , the intensity of the absorption bands at approximately 205 nm and at near 280 nm increased with the increase of the concentration of tropisetron / lansoprazole , indicating the formation of a ground state complex. which could be an evidence of static quenching interaction
Fluorescence spectroscopy measurements:
The purpose of the work was to measure the Fluorescence quenching of the co-solutions of human serum albumin and tropisetron / lansoprazole in phosphate buffered saline (pH = 7.4) at different temperatures .
The fluorescence intensity at λex =289 nm of human serum albumin decreased regularly with increasing the concentration of tropisetron / lansoprazole , while the maximum emission wavelength was almost not changed, suggesting a change in the surrounding environment of the fluorophores (Trp residues in HSA serum albumin) due to binding interaction with tropisetron / lansoprazole and that the binding region of Tropi/ Lanso is mostly in the vicinity of Trp residues. To elucidate the mechanism of fluorescence quenching by which the energy was transferred from human serum albumin to tropi / lanso, the quenching experiments were performed at three temperatures (298 , 303 , 310 0K) and the fluorescence quenching constant (KSV) of bovine serum albumin was calculated using the well-known Stern– Volmer equation which indicated static quenching mechanism rather than a dynamic quenching mechanism.
Moreover, the Binding parameters and identification of the binding site of Tropi / Lanso on human serum albumin, was determined by the modified Stern-Volmer equation. The determined values of Kb indicated that there was a strong binding interaction of Tropi / Lanso with human serum albumin.
In the meantime, decrease in the Kb values was observed with the increase of temperature, suggesting that the stability of the Tropi / Lanso – human serum albumin complex decreased with the increase of temperature.
The competitive displacement experiments:
Identification of the binding sites of Tropi / Lanso on human serum albumin were explored in this part, the competitive displacement experiments were carried out using phenylbutazone , ibuprofen and digoxin as site I , site II and site ІІІ-specific probes and it can be concluded that the binding site of Tropi/ Lanso was mainly located within hydrophobic cavities of subdomain IIA (site I) of human serum albumin.
Thermodynamic parameters and binding mode determination:
Determination of the signs and magnitudes of the thermodynamic parameters were conducted in this part which included the free energy change (ΔG0 ), the enthalpic change (ΔH0 ) and entropic change (ΔS0 ) in the binding process of proteins with drug can be used to confirm the binding modes using Van’t Hoff’s equation
Synchronous fluorescence spectroscopy:
In this part, it was previously reported that when values of Δλ were set at 15 and 60 nm, respectively, it can provide the characteristic information about the microenvironment in a vicinity of disparate chromophores.
The synchronous fluorescence spectra of human serum albumin in the presence of Tropi / Lanso when Δλ were set at 60 nm and 15 nm revealed that the intensity of synchronous fluorescence decreased instantaneously upon addition of Tropi/ Lanso in agreement with the steady-state fluorescence results.
FT-IR analysis:
In this section the conformational change of human serum albumin upon binding to Tropi / Lanso was evaluated by recording FT-IR spectra of free and drug bound human serum albumin in phosphate buffer pH=7.4.
Such as it was previously reported that the IR spectrum of protein exhibits two characteristic absorption bands (amide I and amide II). These are the prominent vibrational bands of the protein backbone. Upon addition of Tropi/ Lanso, a shift in the position of both amide bands was noticed.
Section 5 : chromatography method of tropisetron and Lansoprazole
This section is divided into 3 parts
Part1: Experimental
Part2: Results and Discussion
Part3: application of displacement between Tropi and PPIs
Section 6: voltammetry method
This section is divided into 2 parts
Part1: Experimental
Part2: Results and Discussion
Section 7: docking study
This section is divided into two parts:
Part 1: which includes,
Docking procedure, Study of the binding characteristics to drug site I, Study of the binding characteristics to site II
Part 2: which includes:
1. The validation of the docking setup
2. Application of the validated molecular docking setups
Section 8 :References
Section 9 : English summary
Section 10: Arabic summary.