# Molecular Modeling Studies of Some Uracil and New Deoxyuridine Derivatives

Authors: Yousra Abdel-Mottaleb; M. S. A. Abdel-Mottaleb
Journal: Journal of Chemistry
 (2016)

---

## Abstract

Molecular modeling results reported in this paper are crucial in highlighting the quantitative relationship between the optimized structure and computed molecular properties related to four newly synthesized uracil derivatives with promising biological potential as anticancer bioactive agents. Moreover, 5-fluorouracil (5-FU) and its tautomers and thiouracils molecular properties are studied and correlated with their biological activities. The great medical importance of these and similar molecular systems requires research on their quantitative structure-activity relationships (QSAR) in order to further improve our knowledge about how receptor binding, selectivity, and pharmacological effects are achieved. Modeling is performed in the ground and the first singlet excited states using density functional theory (DFT) and its time-dependent extension (TD-DFT), respectively.


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## Body

## 1. Introduction

Uracils are considered as privileged structures in drug discovery with broad topics of biological activities and synthetic accessibility. Antiviral and anti-tumor are the two most widely reported activities of uracil analogs, however, they also exhibit herbicidal, insecticidal and bactericidal activities. Their antiviral potential is based on the inhibition of a key step in viral replication pathway resulting in potent activities against HIV, hepatitis B and C, the herpes viruses, and so forth. Uracil derivatives such as 5-fluorouracil or 5-chlorouracil were the first pharmacological active derivatives to be generated [1–10]. Numerous modifications of uracil structure (see Figure 1) have been performed to tackle toxicity problems resulting in the development of derivatives possessing better pharmacological and pharmacokinetic properties including increased bioactivity, selectivity, metabolic stability, absorption, and lower toxicity. Researches of new uracils and fused uracil derivatives as bioactive agents are related to modifications of substituents at different positions of the pyrimidine ring [11].Figure 1
Structure and labeling of the molecules studied (blue, gray, light blue, red, and white spheres are representing nitrogen, carbon, fluorine, oxygen, and H atoms, resp.).5-Fluorouracil (5-FU) is an antimetabolite of the pyrimidine analog type and a well-known antitumor agent, which has been widely used in the treatment of solid tumors such as colon or breast cancers [12]. Because 5-FU is similar in shape to uracil but does not perform the same chemistry as uracil, the drug inhibits RNA replication enzymes, thereby eliminating RNA synthesis and stopping the growth of cancerous cells. Although 5-FU has had clinical success as a single agent, it has been modified by different ways to synthesize its derivatives, which may improve its therapeutic index because of its well-known side effects such as short half-life, wide distribution, low selectivity, and various toxic side effects. Recently chemists have paid more attention to the conjugates of 5-FU with a wide spectrum of low- or high-molecular-weight carriers (Figure 2). Generally, the introduction of fluorine atoms into organic molecules usually promotes dramatic changes in their biological properties [13].Figure 2
Mechanism of action of the enzyme TS on dUMP to produce the methylated dTMP leading to DNA generation and cell reproduction. 5-FU is converted into FdUMP in the human body followed by interaction with TS to form a covalent link leading to dysfunction of TS and stopping of DNA generation. As a result, replication and cell division are blocked.Since the target of 5-FU is Thymidylate Synthase (TS), it is of utmost importance to discuss the role of TS enzyme, which catalyzes the conversion of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP) (or 5′-thymidylic acid). It provides the sole de novo pathway for biosynthesis of dTMP (Figure2). It is worth pointing out, however, that fluorinated FdUMP (2′-deoxy-5-fluorouridine 5′-(dihydrogen phosphate)) is generated by conversion of 5-FU in the body. Because of the central role of TS in the synthesis of an essential DNA precursor and its importance as a chemotherapeutic target, the enzyme has been much studied [14, 15].Electronic and structural factors thought of as binding requirements of substrates and analogs will be studied because there are several extremely potent inhibitors of TS. 5-Fluorouracil is considered as mechanism-based inhibitor of TS because it forms stable covalent adduct during TS catalytic function [16].Thus, we are interested in highlighting the relationship between the optimized structure and chemical properties related to the biological activity of the uracils, predominantly for 5-fluorouracil (5-FU), uracil (U), thiouracils (TU), and dithiouracil (DTU) as well as some new uracil derivatives (5′-aminodeoxyuridines (I), (II), and (III)) and amino acid ester conjugate of 5-FU (IV) [12] with promising biological potential. Uridine is a glycosylated pyrimidine analog containing uracil attached to a ribose ring (or more specifically. a ribofuranose) via a β-N1-glycosidic bond.Gaussian 09 and Spartan’14 Modeling packages will be applied using density functional theory (DFT) and its time-dependent extension (TD-DFT) [17–21].Ground- as well as excited-state electronic properties such as optimized geometry parameters and partial atomic charge distribution are obtained for each molecule studied as well as drawings of total electron density mapped with the ESP calculated for surface points only [20]. These drawings provide important visualized information on the molecule as seen by another molecule. Furthermore, reactivity indices (such as chemical potential, global hardness, and softness) of the drug molecules are calculated to map molecular reactivity as inhibitors of Thymidylate Synthase (TS), which plays the central role in the synthesis of an essential DNA precursor, and its importance as a chemotherapeutic target. Furthermore, interaction between 5-FU and the in-body converted form FdUMP will be studied to find out the influence of 5-FU on the reactivity of the substrate FdUMP that enforced TS enzyme to stop functioning.


## 2. Computations

The theoretical calculations are carried out using Gaussian 09, Gaussian Inc. (USA), quantum chemical package within the framework of DFT and TD-DFT (with 6 MOs taken into account in the CI) and visualized by Gausview05 program. 12- and 6-core pro-MAC computers were used to perform the computations. The geometry optimization of each of the studied molecule was carried out in vacuum as implemented in the Gaussian 09 package [22]. The geometry of a molecule was optimized using the hybrid exchange-correlation B3LYP [22] functional with the 3-21G basis set, which is a good compromise between accuracy and efficiency. The further expansion of the basis set has less impact on the accuracy of the molecular parameters. We have also used the 6-31+G(d) basis set for some of the studied species and obtained similar results. Therefore, we will report the results obtained from the 3-21G basis set. Basis set 6-31+G(d,p) was used in case of dUMP and FdUMP molecules.The DFT-B3LYP method has been demonstrated to predict excellent geometries and energies. Vibration frequency calculations were performed to confirm whether the obtained geometry represents a transition or minimum energy structure. The useful reaction field model used for solvation is the conductor-like polarizable continuum model (CPCM), which performs a PCM (polarized continuum model) calculation [22].User-friendly Spartan’14 (parallel suite version, Wavefunction, Inc., USA) computational chemistry software was also used to facilitate our computational tasks. DFT-B3LYP with 6-31G
∗ basis set was used for studying the interactions between 5-FU and FdUMP.


## 3. Results and Discussion

### 3.1. DFT Local Indices for Reactivity and Liability of Electron Density

Local properties are highly desirable in establishing a reactivity-oriented description of molecular systems. Global reactivity indices were estimated according to the equations recommended by Politzer and Murray [23]. In particular, the electronic chemical potentials (μ), chemical hardness (η), and softness (S
=
1
/
η) of the inhibitors studied were evaluated in terms of the one-electron energies of the frontier molecular orbital HOMO and LUMO, using the following equations:(1)
μ
≈
1
2
ϵ
L
U
M
O
+
ϵ
H
O
M
O
,
η
≈
ϵ
L
U
M
O
-
ϵ
H
O
M
O
,where ϵ
H
O
M
O and ϵ
L
U
M
O are the energies of the highest occupied and the lowest unoccupied molecular orbitals, HOMO and LUMO, respectively. The chemical potential μ measures the escaping tendency of electron from equilibrium and the global hardness η can be seen as the resistance to charge transfer (or the band gap as has been shown in Figure 3), while softness S gives indication of how large is electron transfer to/from the molecule when the chemical potential changes [23–25].Figure 3
Representation of chemical potential and molecular hardness and its relationship with HOMO/LUMO.The calculated values are tabulated in Table1. Escaping tendency of electronic charge μ-value slightly increases in solvent cage relative to that computed in vacuum and solvent nature is of minor importance. Chemical potential, which measures the escaping tendency of electrons, and is related to molecular electronegativity (−chemical potential; that is, χ
=
-
μ) decreases in the orderDTU > 5-FU > AMINOFU > TU > U > U124OH-COCH3 > U124F > U124OH,whereas hardness decreases in the order5-FU > U > U124OH > U124OH-COCH3 > U124F > AMINOFU > TU > DTU.Our results are in agreement with the general conclusions that the introduction of fluorine atoms into organic molecules accounts for their remarkable reactivity indices and usually promotes dramatic changes in their biological properties.Table 1
Ground and excited states’ computed reactivity indices values in a.u.∗ of (a) 5-FU, (b) uracil, (c) TU, and (d) DTU in different media.
(a)
Calculated parameters
Vacuum
H
2O
EtOH
DMSO
S
0
S
1
S
0
S
1
S
0
S
1
S
0
S
1
ϵ
L
U
M
O
−0.0326
−0.0724
−0.0332
−0.0747
−0.0332
−0.0442
−0.0333
−0.0443
ϵ
H
O
M
O
−0.2772
−0.2676
−0.2819
−0.2686
−0.2818
−0.2418
−0.2819
−0.2418
−μ
0.155
0.170
0.158
0.172
0.158
0.143
0.158
0.143
η
0.245
0.195
0.249
0.194
0.249
0.198
0.249
0.198
S
∗
4.089
5.124
4.022
5.155
4.023
5.060
4.022
5.062
∗Units: Hartree and Hartree−1 for S
∗.
(b)
Calculated parameters
Vacuum
H
2O
EtOH
DMSO
S
0
S
1
S
0
S
1
S
0
S
1
S
0
S
1
ϵ
L
U
M
O
−0.0251
−0.0625
−0.0298
−0.0651
−0.0295
−0.0649
−0.2977
−0.0650
ϵ
H
O
M
O
−0.2660
−0.2682
−0.2746
−0.2688
−0.2742
−0.2688
−0.2745
−0.2688
−μ
0.146
0.165
0.152
0.167
0.152
0.167
0.152
0.167
η
0.241
0.206
0.245
0.204
0.245
0.204
0.245
0.204
S
∗
4.152
4.863
4.085
4.907
4.086
4.904
4.086
4.902
(c)
Calculated parameters
Vacuum
H
2O
EtOH
DMSO
S
0
S
1
S
0
S
1
S
0
S
1
S
0
S
1
ϵ
L
U
M
O
−0.7711
−0.08705
−0.08759
−0.08860
−0.08709
−0.08853
−0.08743
−0.08858
ϵ
H
O
M
O
−0.21387
−0.23176
−0.22459
−0.22965
−0.22408
−0.22975
−0.22443
−0.22969
−μ
0.146
0.159
0.156
0.159
0.156
0.59
0.156
0.159
η
0.137
0.145
0.137
0.141
0.137
0.141
0.137
0.141
S
∗
7.312
6.910
7.300
7.087
7.300
7.081
7.300
7.087
(d)
Calculated parameters
Vacuum
H
2O
EtOH
DMSO
S
0
S
1
S
0
S
1
S
0
S
1
S
0
S
1
ϵ
L
U
M
O
−0.9777
−0.0927
−0.1035
−0.0947
−0.1033
−0.0946
−0.1035
−0.0947
ϵ
H
O
M
O
−0.2186
−0.2354
−0.2772
−0.2342
−0.2268
−0.2342
−0.2270
−0.2342
−μ
0.158
0.164
0.165
0.161
0.161
0.164
0.165
0.165
η
0.121
0.143
0.124
0.139
0.124
0.140
0.124
0.140
S
∗
8.267
7.010
8.088
7.171
8.900
7.163
8.092
7.169
(e)
Calculated parameters
U124 X=OH. R=OCCH3 (I)aminodeoxyuridine
U124 X=OH. R=H (II)deoxyuridine
U124 X=F. R=H (III)uridine
Amino-FU (IV)5-FU-amino acid ester conjugate
S
0
S
1
S
0
S
1
S
0
S
1
S
0
ϵ
L
U
M
O
−0.02308
−0.05903
−0.01683
−0.05338
−0.02104
−0.05322
−0.03273
ϵ
H
O
M
O
−0.26161
−0.26023
−0.25667
−0.25404
−0.25764
−0.25585
−0.26791
−μ
0.142
0.160
0.137
0.154
0.139
0.155
0.150
η
0.239
0.201
0.240
0.201
0.237
0.203
0.235
S
∗
4.192
4.970
4.169
4.983
4.227
4.935
4.242The calculated reactivity descriptors listed in Table1(a) favor least charge transfer liability in case of 5-FU. Electronic and structural factors thought of as binding requirements of substrates and analogs studied could shed light on its role as potent inhibitors of TS. Our results explain why 5-FU is previously considered as mechanism-based inhibitor of TS because it forms stable covalent adduct during TS catalytic function [16].Further support comes from the computed Mulliken charges on individual atoms of molecules (see Supplementary Table  1 of the Supplementary Material available online athttp://dx.doi.org/10.1155/2016/5134732). Inspection of Supplementary Table 1 (Supplementary Material) reveals generally the following.(1) Nitrogen and oxygen atoms have the majority of negative charges in all molecules studied, whereas C4 of the uracil ring (tagged 5 in case of 5-FU) has the majority of positive charges.(2) They increase in charges due to solvation.(3) The general sequence of decreasing charges on the simple uracils is 5-FU > U > TU > DTU.And in case of 5′-amino-2′-deoxyuridines and the amino acid ester conjugate of 5-FU (IV) order is I ≥ II > III > IV.(4) Remarkable charge alternation is observed in case of 5-FU gas phase, whereas solvation induces charge equalization on different atoms.


### 3.2. Analysis of Mapped Molecular Electrostatic Surface Potential (ESP)

The molecular electrostatic potential surface (ESP) provides a visual method to understand the relative polarity of the compounds [23, 24]. Electrostatic potential surfaces mapped (ESP map) with electron density are shown in Figure 4 and Figure 5 illustrates the charge distributions of the molecule three dimensionally and is very useful in research of molecular structure with its physiochemical property relationship [25–27].Figure 4
ESP map of simple uraciles. Isovalue = 0.0004; ESP value ranges between −175 kJmol−1 (red) and +279 kJmol−1 (blue).Figure 5
Transparent ESP map of uredines (I, II, and III) and amino acid ester of 5-FU (IV). Isovalue = 0.0004; ESP value ranges between −140 kJmol−1 (red) and +140 kJmol−1 (blue).These pictures illustrate an electrostatic potential model of the compounds, computed at the 0.0004 a.u. isodensity surface. ESP maps will be used to determine how molecules interact with one another (see Section3.3).


### 3.3. The Interaction between 5-FU and Its 5-Fluoro-2′-deoxyuridine 5′-Monophosphate (FdUMP)

Administered, as the premetabolite, 5-FU is a mechanism-based inhibitor of TS that is used in chemotherapy.We used Spartan’14 parallel suite (8 threads, 4-core Intel i7-based processor) Mac version to simplify exploration of the effect of 5-FU administered on the FdUMP produced in human body. This has a direct impact on TS suicide substrate leading to a synergistic effect on blocking cell division and replication. Based on the ESP maps, we examined different orientations for steric fit to FdUMP with almost the same results as reflected in the partial atomic charges. We scored each on the basis of the equilibrium geometry minimization as well as the energy of the electrostatic interaction with the substrates of the enzyme. Global reactivity descriptors, chemical potential, hardness, and softness beside partial atomic charges as well as ESP maps of the molecules, are calculated (Tables2 and 3) and visualized in Figure 6. The interaction between 5-FU and FdUMP results in the following. (1) Two H-bonds form between H2⋯O8 and H22⋯O5 with optimized bond lengths 1.784 and 1.837 Å. (2) Molecular electronegativity (expressed as increasing chemical potential) increases due to charge transfer from 5-FU part to FdUMP moiety (see HOMO-LUMO charge transfer in Figure 7) concomitant with decrease in global hardness and increase in global softness. Thus, it could be reasonably predicted that these changes should result in more reactive adduct when covalently combined with TS, as suggested by the mechanism of TS function, leading to faster stop action of TS enzyme in producing DNA. As a result, cell division and DNA replication should be remarkably blocked. (3) Atomic charges remarkably changed, in particular, on the interacting sites of the molecular fragments. It is worth pointing out that the nonconverted portions of the 5-FU enhance the reactivity of the converted 5-FU to FdUMP towards the TS enzyme leading to efficient blocking of cell division and DNA replication.Table 2
Reactivity descriptors for the complex resulting from the interactions between 5-FU and the human body generated 5-F-2′-dUMP (values are in eV).
Calculated parameters
5-FU
5-F-2′-dUMP
5-F-2′-dUMP + 5FU complex
ϵ
L
U
M
O
−1.38
−1.27
−1.53
ϵ
H
O
M
O
−6.79
−6.59
−6.58
-
μ
4.085
3.93
4.055
η
5.41
5.32
5.05
S
∗
0.185
0.188
0.198
∗eV−1.Table 3
Computed partial atomic charges on individual molecules and on the adduct resulting from the interactions between 5-FU and the in-body produced FdUMP enzyme suicide agent of TS enzyme (ES = electrostatic, M = Mulliken, and N = natural charge).
Mol.
Number
Atomlabel
Atomic charges on the individual drugsFdUMP and 5FU
Number
Atomlabel
Atomic charges on the adduct5FU-FdUMP
ES
M
N
ES
M
N
FdUMP
1
N1
−0.186
−0.54
−0.471
1
N1
−0.073
−0.542
−0.476
2
C2
0.615
0.805
0.828
2
C2
0.607
0.794
0.827
3
N3
−0.566
−0.723
−0.672
3
N3
−0.582
−0.758
−0.668
4
C4
0.577
0.599
0.608
4
C4
0.541
0.622
0.62
5
C5
0.017
0.253
0.259
5
C5
0.087
0.257
0.261
6
C6
−0.12
0.033
−0.035
6
C6
−0.228
0.037
−0.02
7
O7
−0.529
−0.513
−0.624
7
O7
−0.505
−0.505
−0.617
8
O8
−0.475
−0.483
−0.571
8
O8
−0.513
−0.545
−0.639
9
F9
−0.143
−0.292
−0.33
9
F1
−0.154
−0.291
−0.329
10
C10
0.379
0.312
0.288
10
C10
0.317
0.295
0.292
11
C11
−0.288
−0.322
−0.519
11
C11
−0.56
−0.337
−0.524
12
C12
0.122
0.087
0.074
12
C12
0.43
0.087
0.069
13
C13
0.13
0.112
0.04
13
C13
0.179
0.127
0.047
14
O14
−0.558
−0.623
−0.751
14
O14
−0.67
−0.629
−0.751
15
O15
−0.413
−0.52
−0.593
15
O15
−0.426
−0.522
−0.592
16
C16
−0.138
−0.064
−0.13
16
C16
−0.161
−0.059
−0.122
17
O17
−0.315
−0.537
−0.871
17
O17
−0.344
−0.536
−0.874
18
P18
1.094
1.178
2.565
18
P18
1.126
1.162
2.565
19
O19
−0.614
−0.517
−1.052
19
O19
−0.627
−0.515
−1.054
20
O20
−0.613
−0.692
−1.037
20
O20
−0.616
−0.684
−1.034
21
O21
−0.597
−0.653
−1.018
21
O21
−0.598
−0.653
−1.019
22
H22
0.371
0.363
0.453
22
H22
0.403
0.42
0.476
23
H23
0.209
0.216
0.27
23
H23
0.242
0.227
0.274
24
H24
0.08
0.189
0.257
24
H24
0.087
0.186
0.255
25
H25
0.116
0.192
0.269
25
H25
0.18
0.192
0.271
26
H26
0.158
0.201
0.282
26
H26
0.187
0.174
0.261
27
H27
0.085
0.15
0.231
27
H27
0.01
0.172
0.236
28
H28
0.092
0.147
0.246
28
H28
0.099
0.165
0.249
29
H29
0.383
0.395
0.475
29
H29
0.421
0.406
0.485
30
H30
0.12
0.169
0.235
30
H30
0.136
0.184
0.245
31
H31
0.151
0.18
0.237
31
H31
0.161
0.183
0.241
32
H32
0.418
0.452
0.529
32
H32
0.417
0.449
0.527
33
H33
0.439
0.446
0.526
33
H33
0.438
0.448
0.527
5-FU
1
F0
−0.188
−0.387
−0.322
34
F0
−0.134
−0.274
−0.314
2
C1
−0.053
0.218
0.223
35
C1
0.043
0.278
0.275
3
C2
0.006
0.182
0.015
36
C3
−0.087
0.031
−0.047
4
N3
−0.683
−1.002
−0.761
37
N2
−0.407
−0.678
−0.633
5
C4
1.057
1.216
0.984
38
C7
0.652
0.79
0.832
6
O5
−0.664
−0.619
−0.658
39
O5
−0.565
−0.539
−0.668
7
N6
−0.884
−1.047
−0.813
40
N6
−0.539
−0.696
−0.67
8
C7
0.905
0.889
0.719
41
C8
0.578
0.58
0.604
9
O8
−0.581
−0.586
−0.592
42
O1
−0.479
−0.481
−0.57
10
H1
0.237
0.32
0.272
43
H1
0.199
0.199
0.261
11
H2
0.412
0.404
0.463
44
H2
0.365
0.418
0.47
12
H3
0.436
0.411
0.468
45
H3
0.363
0.363
0.453Figure 6
ESP map of FdUMP (a), 5-FU (b), the adduct of (a) and (b) (c), the full ESP map of the adduct (d), and the formed two H-bonds between (a) and (b) (e).
(a)
(b)
(c)
(d)
(e)Figure 7
Frontier MOs of 5-FU (a), FdUMP (b), and the complex adduct of 5-FU and FdUMP (c) showing the charge transfer nature of the transition HOMO (mainly on 5-FU fragment to LUMO localized on FdUMP fragment of the complex).
(a)
(b)
(c)


### 3.4. Tautomers Stabilities

In a previous study [28] it was confirmed that the diketo tautomer is the most stable form of uracil and its derivatives in a water solution. In solid state 5-FU exists also in the ketonic form [29]. Tables 4(a) and 4(b) and Figure 8 list the all possible tautomers that are generated using Spartan’14 tautomer search and optimized by B3LYP/6-31G
∗ model. In agreement with the aforementioned studies, our thermodynamic data indicates the stability of ketonic form (T6) relative to other forms. Examining frontier orbitals reveals that T2 with lowest energy gap is the most reactive form. The least energy and thermodynamically stable T3 that exhibits largest dipole moment shows substantial solvation energy change in aqueous solution.Table 4
(a) Solvation energy, frontier orbital energies, dipole moment, and zero-point energy of the six possible tautomers of 5-FU (see Figure8). (b) Total energies in the gas phase (E) and aqueous solution (E
a
q) in Hartree (atomic unit) and thermodynamic parameters of the six possible tautomers of 5-FU.
(a)
Tautomer
SolvationE (kJ/mol)
E HOMO (eV)
E LUMO (eV)
Dipole(debye)
ZPE(kJ/mol)
T6
−38.92
−6.79
−1.38
3.90
207.48
T1
−80.48
−6.16
−1.26
5.03
200.36
T5
−105.10
−5.80
−1.02
3.41
192.36
T2
−105.35
−6.05
−2.04
6.84
198.77
T4
−110.20
−6.29
−1.82
5.30
199.15
T3
−142.32
−6.34
−0.96
8.92
199.35
(b)
Tautomer
E (au)
E
a
q (au)
H
o (au)
S
o (J/molo)
G
o (au)
T6
−514.035880
−514.050703
−513.949227
342.8
−513.988156
T1
−513.948808
−514.003016
−513.882627
352.01
−513.922601
T5
−513.956560
−513.998531
−513.825376
361.45
−513.866422
T2
−513.967035
−513.997688
−513.867448
352.56
−513.907485
T4
−513.951220
−513.991344
−513.872636
352.57
−513.912674
T3
−513.907176
−513.947206
−513.864678
353.32
−513.90480Figure 8
The six possible tautomers of 5-FU.


## 3.1. DFT Local Indices for Reactivity and Liability of Electron Density

Local properties are highly desirable in establishing a reactivity-oriented description of molecular systems. Global reactivity indices were estimated according to the equations recommended by Politzer and Murray [23]. In particular, the electronic chemical potentials (μ), chemical hardness (η), and softness (S
=
1
/
η) of the inhibitors studied were evaluated in terms of the one-electron energies of the frontier molecular orbital HOMO and LUMO, using the following equations:(1)
μ
≈
1
2
ϵ
L
U
M
O
+
ϵ
H
O
M
O
,
η
≈
ϵ
L
U
M
O
-
ϵ
H
O
M
O
,where ϵ
H
O
M
O and ϵ
L
U
M
O are the energies of the highest occupied and the lowest unoccupied molecular orbitals, HOMO and LUMO, respectively. The chemical potential μ measures the escaping tendency of electron from equilibrium and the global hardness η can be seen as the resistance to charge transfer (or the band gap as has been shown in Figure 3), while softness S gives indication of how large is electron transfer to/from the molecule when the chemical potential changes [23–25].Figure 3
Representation of chemical potential and molecular hardness and its relationship with HOMO/LUMO.The calculated values are tabulated in Table1. Escaping tendency of electronic charge μ-value slightly increases in solvent cage relative to that computed in vacuum and solvent nature is of minor importance. Chemical potential, which measures the escaping tendency of electrons, and is related to molecular electronegativity (−chemical potential; that is, χ
=
-
μ) decreases in the orderDTU > 5-FU > AMINOFU > TU > U > U124OH-COCH3 > U124F > U124OH,whereas hardness decreases in the order5-FU > U > U124OH > U124OH-COCH3 > U124F > AMINOFU > TU > DTU.Our results are in agreement with the general conclusions that the introduction of fluorine atoms into organic molecules accounts for their remarkable reactivity indices and usually promotes dramatic changes in their biological properties.Table 1
Ground and excited states’ computed reactivity indices values in a.u.∗ of (a) 5-FU, (b) uracil, (c) TU, and (d) DTU in different media.
(a)
Calculated parameters
Vacuum
H
2O
EtOH
DMSO
S
0
S
1
S
0
S
1
S
0
S
1
S
0
S
1
ϵ
L
U
M
O
−0.0326
−0.0724
−0.0332
−0.0747
−0.0332
−0.0442
−0.0333
−0.0443
ϵ
H
O
M
O
−0.2772
−0.2676
−0.2819
−0.2686
−0.2818
−0.2418
−0.2819
−0.2418
−μ
0.155
0.170
0.158
0.172
0.158
0.143
0.158
0.143
η
0.245
0.195
0.249
0.194
0.249
0.198
0.249
0.198
S
∗
4.089
5.124
4.022
5.155
4.023
5.060
4.022
5.062
∗Units: Hartree and Hartree−1 for S
∗.
(b)
Calculated parameters
Vacuum
H
2O
EtOH
DMSO
S
0
S
1
S
0
S
1
S
0
S
1
S
0
S
1
ϵ
L
U
M
O
−0.0251
−0.0625
−0.0298
−0.0651
−0.0295
−0.0649
−0.2977
−0.0650
ϵ
H
O
M
O
−0.2660
−0.2682
−0.2746
−0.2688
−0.2742
−0.2688
−0.2745
−0.2688
−μ
0.146
0.165
0.152
0.167
0.152
0.167
0.152
0.167
η
0.241
0.206
0.245
0.204
0.245
0.204
0.245
0.204
S
∗
4.152
4.863
4.085
4.907
4.086
4.904
4.086
4.902
(c)
Calculated parameters
Vacuum
H
2O
EtOH
DMSO
S
0
S
1
S
0
S
1
S
0
S
1
S
0
S
1
ϵ
L
U
M
O
−0.7711
−0.08705
−0.08759
−0.08860
−0.08709
−0.08853
−0.08743
−0.08858
ϵ
H
O
M
O
−0.21387
−0.23176
−0.22459
−0.22965
−0.22408
−0.22975
−0.22443
−0.22969
−μ
0.146
0.159
0.156
0.159
0.156
0.59
0.156
0.159
η
0.137
0.145
0.137
0.141
0.137
0.141
0.137
0.141
S
∗
7.312
6.910
7.300
7.087
7.300
7.081
7.300
7.087
(d)
Calculated parameters
Vacuum
H
2O
EtOH
DMSO
S
0
S
1
S
0
S
1
S
0
S
1
S
0
S
1
ϵ
L
U
M
O
−0.9777
−0.0927
−0.1035
−0.0947
−0.1033
−0.0946
−0.1035
−0.0947
ϵ
H
O
M
O
−0.2186
−0.2354
−0.2772
−0.2342
−0.2268
−0.2342
−0.2270
−0.2342
−μ
0.158
0.164
0.165
0.161
0.161
0.164
0.165
0.165
η
0.121
0.143
0.124
0.139
0.124
0.140
0.124
0.140
S
∗
8.267
7.010
8.088
7.171
8.900
7.163
8.092
7.169
(e)
Calculated parameters
U124 X=OH. R=OCCH3 (I)aminodeoxyuridine
U124 X=OH. R=H (II)deoxyuridine
U124 X=F. R=H (III)uridine
Amino-FU (IV)5-FU-amino acid ester conjugate
S
0
S
1
S
0
S
1
S
0
S
1
S
0
ϵ
L
U
M
O
−0.02308
−0.05903
−0.01683
−0.05338
−0.02104
−0.05322
−0.03273
ϵ
H
O
M
O
−0.26161
−0.26023
−0.25667
−0.25404
−0.25764
−0.25585
−0.26791
−μ
0.142
0.160
0.137
0.154
0.139
0.155
0.150
η
0.239
0.201
0.240
0.201
0.237
0.203
0.235
S
∗
4.192
4.970
4.169
4.983
4.227
4.935
4.242The calculated reactivity descriptors listed in Table1(a) favor least charge transfer liability in case of 5-FU. Electronic and structural factors thought of as binding requirements of substrates and analogs studied could shed light on its role as potent inhibitors of TS. Our results explain why 5-FU is previously considered as mechanism-based inhibitor of TS because it forms stable covalent adduct during TS catalytic function [16].Further support comes from the computed Mulliken charges on individual atoms of molecules (see Supplementary Table  1 of the Supplementary Material available online athttp://dx.doi.org/10.1155/2016/5134732). Inspection of Supplementary Table 1 (Supplementary Material) reveals generally the following.(1) Nitrogen and oxygen atoms have the majority of negative charges in all molecules studied, whereas C4 of the uracil ring (tagged 5 in case of 5-FU) has the majority of positive charges.(2) They increase in charges due to solvation.(3) The general sequence of decreasing charges on the simple uracils is 5-FU > U > TU > DTU.And in case of 5′-amino-2′-deoxyuridines and the amino acid ester conjugate of 5-FU (IV) order is I ≥ II > III > IV.(4) Remarkable charge alternation is observed in case of 5-FU gas phase, whereas solvation induces charge equalization on different atoms.


## 3.2. Analysis of Mapped Molecular Electrostatic Surface Potential (ESP)

The molecular electrostatic potential surface (ESP) provides a visual method to understand the relative polarity of the compounds [23, 24]. Electrostatic potential surfaces mapped (ESP map) with electron density are shown in Figure 4 and Figure 5 illustrates the charge distributions of the molecule three dimensionally and is very useful in research of molecular structure with its physiochemical property relationship [25–27].Figure 4
ESP map of simple uraciles. Isovalue = 0.0004; ESP value ranges between −175 kJmol−1 (red) and +279 kJmol−1 (blue).Figure 5
Transparent ESP map of uredines (I, II, and III) and amino acid ester of 5-FU (IV). Isovalue = 0.0004; ESP value ranges between −140 kJmol−1 (red) and +140 kJmol−1 (blue).These pictures illustrate an electrostatic potential model of the compounds, computed at the 0.0004 a.u. isodensity surface. ESP maps will be used to determine how molecules interact with one another (see Section3.3).


## 3.3. The Interaction between 5-FU and Its 5-Fluoro-2′-deoxyuridine 5′-Monophosphate (FdUMP)

Administered, as the premetabolite, 5-FU is a mechanism-based inhibitor of TS that is used in chemotherapy.We used Spartan’14 parallel suite (8 threads, 4-core Intel i7-based processor) Mac version to simplify exploration of the effect of 5-FU administered on the FdUMP produced in human body. This has a direct impact on TS suicide substrate leading to a synergistic effect on blocking cell division and replication. Based on the ESP maps, we examined different orientations for steric fit to FdUMP with almost the same results as reflected in the partial atomic charges. We scored each on the basis of the equilibrium geometry minimization as well as the energy of the electrostatic interaction with the substrates of the enzyme. Global reactivity descriptors, chemical potential, hardness, and softness beside partial atomic charges as well as ESP maps of the molecules, are calculated (Tables2 and 3) and visualized in Figure 6. The interaction between 5-FU and FdUMP results in the following. (1) Two H-bonds form between H2⋯O8 and H22⋯O5 with optimized bond lengths 1.784 and 1.837 Å. (2) Molecular electronegativity (expressed as increasing chemical potential) increases due to charge transfer from 5-FU part to FdUMP moiety (see HOMO-LUMO charge transfer in Figure 7) concomitant with decrease in global hardness and increase in global softness. Thus, it could be reasonably predicted that these changes should result in more reactive adduct when covalently combined with TS, as suggested by the mechanism of TS function, leading to faster stop action of TS enzyme in producing DNA. As a result, cell division and DNA replication should be remarkably blocked. (3) Atomic charges remarkably changed, in particular, on the interacting sites of the molecular fragments. It is worth pointing out that the nonconverted portions of the 5-FU enhance the reactivity of the converted 5-FU to FdUMP towards the TS enzyme leading to efficient blocking of cell division and DNA replication.Table 2
Reactivity descriptors for the complex resulting from the interactions between 5-FU and the human body generated 5-F-2′-dUMP (values are in eV).
Calculated parameters
5-FU
5-F-2′-dUMP
5-F-2′-dUMP + 5FU complex
ϵ
L
U
M
O
−1.38
−1.27
−1.53
ϵ
H
O
M
O
−6.79
−6.59
−6.58
-
μ
4.085
3.93
4.055
η
5.41
5.32
5.05
S
∗
0.185
0.188
0.198
∗eV−1.Table 3
Computed partial atomic charges on individual molecules and on the adduct resulting from the interactions between 5-FU and the in-body produced FdUMP enzyme suicide agent of TS enzyme (ES = electrostatic, M = Mulliken, and N = natural charge).
Mol.
Number
Atomlabel
Atomic charges on the individual drugsFdUMP and 5FU
Number
Atomlabel
Atomic charges on the adduct5FU-FdUMP
ES
M
N
ES
M
N
FdUMP
1
N1
−0.186
−0.54
−0.471
1
N1
−0.073
−0.542
−0.476
2
C2
0.615
0.805
0.828
2
C2
0.607
0.794
0.827
3
N3
−0.566
−0.723
−0.672
3
N3
−0.582
−0.758
−0.668
4
C4
0.577
0.599
0.608
4
C4
0.541
0.622
0.62
5
C5
0.017
0.253
0.259
5
C5
0.087
0.257
0.261
6
C6
−0.12
0.033
−0.035
6
C6
−0.228
0.037
−0.02
7
O7
−0.529
−0.513
−0.624
7
O7
−0.505
−0.505
−0.617
8
O8
−0.475
−0.483
−0.571
8
O8
−0.513
−0.545
−0.639
9
F9
−0.143
−0.292
−0.33
9
F1
−0.154
−0.291
−0.329
10
C10
0.379
0.312
0.288
10
C10
0.317
0.295
0.292
11
C11
−0.288
−0.322
−0.519
11
C11
−0.56
−0.337
−0.524
12
C12
0.122
0.087
0.074
12
C12
0.43
0.087
0.069
13
C13
0.13
0.112
0.04
13
C13
0.179
0.127
0.047
14
O14
−0.558
−0.623
−0.751
14
O14
−0.67
−0.629
−0.751
15
O15
−0.413
−0.52
−0.593
15
O15
−0.426
−0.522
−0.592
16
C16
−0.138
−0.064
−0.13
16
C16
−0.161
−0.059
−0.122
17
O17
−0.315
−0.537
−0.871
17
O17
−0.344
−0.536
−0.874
18
P18
1.094
1.178
2.565
18
P18
1.126
1.162
2.565
19
O19
−0.614
−0.517
−1.052
19
O19
−0.627
−0.515
−1.054
20
O20
−0.613
−0.692
−1.037
20
O20
−0.616
−0.684
−1.034
21
O21
−0.597
−0.653
−1.018
21
O21
−0.598
−0.653
−1.019
22
H22
0.371
0.363
0.453
22
H22
0.403
0.42
0.476
23
H23
0.209
0.216
0.27
23
H23
0.242
0.227
0.274
24
H24
0.08
0.189
0.257
24
H24
0.087
0.186
0.255
25
H25
0.116
0.192
0.269
25
H25
0.18
0.192
0.271
26
H26
0.158
0.201
0.282
26
H26
0.187
0.174
0.261
27
H27
0.085
0.15
0.231
27
H27
0.01
0.172
0.236
28
H28
0.092
0.147
0.246
28
H28
0.099
0.165
0.249
29
H29
0.383
0.395
0.475
29
H29
0.421
0.406
0.485
30
H30
0.12
0.169
0.235
30
H30
0.136
0.184
0.245
31
H31
0.151
0.18
0.237
31
H31
0.161
0.183
0.241
32
H32
0.418
0.452
0.529
32
H32
0.417
0.449
0.527
33
H33
0.439
0.446
0.526
33
H33
0.438
0.448
0.527
5-FU
1
F0
−0.188
−0.387
−0.322
34
F0
−0.134
−0.274
−0.314
2
C1
−0.053
0.218
0.223
35
C1
0.043
0.278
0.275
3
C2
0.006
0.182
0.015
36
C3
−0.087
0.031
−0.047
4
N3
−0.683
−1.002
−0.761
37
N2
−0.407
−0.678
−0.633
5
C4
1.057
1.216
0.984
38
C7
0.652
0.79
0.832
6
O5
−0.664
−0.619
−0.658
39
O5
−0.565
−0.539
−0.668
7
N6
−0.884
−1.047
−0.813
40
N6
−0.539
−0.696
−0.67
8
C7
0.905
0.889
0.719
41
C8
0.578
0.58
0.604
9
O8
−0.581
−0.586
−0.592
42
O1
−0.479
−0.481
−0.57
10
H1
0.237
0.32
0.272
43
H1
0.199
0.199
0.261
11
H2
0.412
0.404
0.463
44
H2
0.365
0.418
0.47
12
H3
0.436
0.411
0.468
45
H3
0.363
0.363
0.453Figure 6
ESP map of FdUMP (a), 5-FU (b), the adduct of (a) and (b) (c), the full ESP map of the adduct (d), and the formed two H-bonds between (a) and (b) (e).
(a)
(b)
(c)
(d)
(e)Figure 7
Frontier MOs of 5-FU (a), FdUMP (b), and the complex adduct of 5-FU and FdUMP (c) showing the charge transfer nature of the transition HOMO (mainly on 5-FU fragment to LUMO localized on FdUMP fragment of the complex).
(a)
(b)
(c)


## 3.4. Tautomers Stabilities

In a previous study [28] it was confirmed that the diketo tautomer is the most stable form of uracil and its derivatives in a water solution. In solid state 5-FU exists also in the ketonic form [29]. Tables 4(a) and 4(b) and Figure 8 list the all possible tautomers that are generated using Spartan’14 tautomer search and optimized by B3LYP/6-31G
∗ model. In agreement with the aforementioned studies, our thermodynamic data indicates the stability of ketonic form (T6) relative to other forms. Examining frontier orbitals reveals that T2 with lowest energy gap is the most reactive form. The least energy and thermodynamically stable T3 that exhibits largest dipole moment shows substantial solvation energy change in aqueous solution.Table 4
(a) Solvation energy, frontier orbital energies, dipole moment, and zero-point energy of the six possible tautomers of 5-FU (see Figure8). (b) Total energies in the gas phase (E) and aqueous solution (E
a
q) in Hartree (atomic unit) and thermodynamic parameters of the six possible tautomers of 5-FU.
(a)
Tautomer
SolvationE (kJ/mol)
E HOMO (eV)
E LUMO (eV)
Dipole(debye)
ZPE(kJ/mol)
T6
−38.92
−6.79
−1.38
3.90
207.48
T1
−80.48
−6.16
−1.26
5.03
200.36
T5
−105.10
−5.80
−1.02
3.41
192.36
T2
−105.35
−6.05
−2.04
6.84
198.77
T4
−110.20
−6.29
−1.82
5.30
199.15
T3
−142.32
−6.34
−0.96
8.92
199.35
(b)
Tautomer
E (au)
E
a
q (au)
H
o (au)
S
o (J/molo)
G
o (au)
T6
−514.035880
−514.050703
−513.949227
342.8
−513.988156
T1
−513.948808
−514.003016
−513.882627
352.01
−513.922601
T5
−513.956560
−513.998531
−513.825376
361.45
−513.866422
T2
−513.967035
−513.997688
−513.867448
352.56
−513.907485
T4
−513.951220
−513.991344
−513.872636
352.57
−513.912674
T3
−513.907176
−513.947206
−513.864678
353.32
−513.90480Figure 8
The six possible tautomers of 5-FU.


## 4. Conclusion

Reactivity descriptors and electrostatic surface potential mapped onto total electron density (ESP map) of eight uracil derivatives, namely, 5-fluorouracil (5-FU), uracil (U), thiouracils (TU), and dithiouracil (DTU) as well as some new uracil derivatives (5′-aminodeoxyuredines I, II, and III) and amino acid ester conjugate of 5-FU (IV), have been calculated and visualized. The results revealed the following.(a) Calculated molecular electronegativity (−chemical potential; that is,χ
=
-
μ) decreases in the order DTU > 5-FU > IV > TU > U > I > III > II.(b) On the other hand, hardness, which measures the resistance to electron transfer, is closely linked to band gap. It decreases in the order 5-FU > U > II > I > III > IV > TU > DTU.Our results are in agreement with the general conclusions that the introduction of fluorine atoms into uracil rings accounts for their remarkable reactivity indices and usually promotes dramatic changes in their biological properties.(c) The calculated reactivity descriptors favor least charge transfer liability in case of 5-FU and the general sequence of decreasing Mulliken charges on the simple uracils is 5-FU > U > TU > DTU.(d) And in case of 5′-amino-2′-deoxyuridines and the amino acid ester conjugate of 5-FU (IV) order is I ≥ II > III > IV.(e) Remarkable charge alternation is observed in case of 5-FU gas phase, whereas solvation induces charge equalization on different atoms.(f) As can be seen from the ESP maps of the studied molecules, while regions having the negative potential are over the oxygen atoms of the uracil part of the molecules; the regions having the zero or small positive potential are over the remaining part. These results provide a visualized recognition tool for the studied molecules, guiding its reactivity towards receptors.(g) The interaction between 5-FU and FdUMP results in a synergistic effect in blocking cell division and DNA replications.Finally, amongst the tautomers of 5-FU, the diketo form is the most stable one. Molecular modeling, thus, is accelerating the pace of cancer drug efficiency and discovery.



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*Source: 5134732-2016-08-28.xml*