Pyridine-Ureas as Potential Anticancer Agents: Synthesis and In Vitro Biological Evaluation

In our endeavor towards the development of effective anticancer agents, a novel series of pyridine-ureas 8a–n were synthesized. All the newly prepared derivatives were evaluated in vitro for their growth inhibitory activity towards the proliferation of breast cancer MCF-7 cell line. Compounds 8e and 8n were found to be the most active congeners against MCF-7 cells (IC50 = 0.22 and 1.88 µM after 48 h treatment; 0.11 and 0.80 µM after 72 h treatment, respectively) with increased activity compared to the reference drug doxorubicin (IC50 = 1.93 µM). Moreover, eight selected pyridines 8b, 8d, 8e, 8i, 8j and 8l–n were evaluated for their in vitro anticancer activity according to the US-NCI protocol. Pyridines 8b and 8e proved to be the most effective anticancer agents in the NCI assay with mean inhibition = 43 and 49%, respectively. Both 8b and 8e exhibited anti-proliferative activity against all tested cancer cell lines from all subpanels growth inhibition (GI for 8b; 12–78%, GI for 8e; 15–91%). Pyridines 8b and 8e were screened in vitro for their inhibitory activity against VEGFR-2. Both compounds inhibited VEGFR-2 at micromolar IC50 values 5.0 ± 1.91 and 3.93 ± 0.73 µM, respectively. The most active pyridines were filtered according to the Lipinski and Veber rules and all of them passed these filters. Finally, several ADME descriptors were predicted for the active pyridines through a theoretical kinetic study.


Introduction
Cancer represents one of the most important health problems worldwide because it is deemed to be the second major cause of mortality throughout the world after cardiovascular diseases.The high mortality rate that occurs in cancer patients is due to the late diagnosis, and consequently the delayed initiation of the medical treatment of the disease.Although there are a large number of chemotherapeutic drugs available, the medical need is still largely unmet.According to the latest GLOBOCAN statistics, about 14.1 million newly diagnosed cancer cases were recorded.Among the newly diagnosed cases in females, breast cancer ranked in the first position worldwide and was regarded as the main cause of deaths attributed to cancer [1,2].Accordingly, there is a pressing necessity to pay much effort to modify drug leads from the point of view of drug design to afford new bioactive chemical entities that offer improvements over current therapies.
In the current medical era, non-fused pyridines have emerged as a substantial class of heterocycles, which are endowed with diverse biological activities, chiefly anticancer activities [3][4][5].Several pyridine-based small molecules have been approved as anticancer drugs, to name just a few: Sorafenib I (Nexavar ® , Figure 1), Regorafenib II (Stivarga ® , Figure 1), Vismodegib III (Erivedge ® , Figure 1) and Crizotinib IV (Xalkori ® , Figure 1) [6][7][8].Subsequently, extensive efforts have been devoted to develop several pyridine-based derivatives as effective anticancer agents.BRN-103 V (Figure 1) is a nicotinamide derivative that suppresses the VEGF-induced phosphorylation of VEGFR-2 and the activation of AKT and eNOS.Also, it inhibits VEGF-induced migration, proliferation and capillary-like tube formation of HUVECs [9].SKLB610 VI (Figure 1) is a multi-targeted kinase inhibitor activity towards VEGFR-2 and FGFR-2.SKLB610 has anti-proliferative effects, especially on human colorectal cancer cell line HCT-116 and human NSCLC cell line A549, with significant activity on tumor xenografts in nude mice without evident toxicity [10].
On the other hand, urea derivatives represent one of the most useful classes of anticancer agents, with a wide range of activities towards various tumors [11].Urea functionality is the main pharmacophoric feature in several anticancer drugs such as Sorafenib I and Regorafenib II.Linifanib VII (Figure 1), an ureido indazole derivative, has selective inhibitory activity towards VEGFR and PDGFRs.Linifanib is in phase II clinical trials for patients with locally advanced or metastatic non-small cell lung cancer [12].SLC-0111 VIII (Figure 1), an ureido benzenesulfonamide derivative, is currently in phase I/II clinical trials as a potential anticancer drug.SLC-0111 exhibited selectivity towards inhibition of the transmembrane isoforms human carbonic anhydrase (hCA) IX/XII (over the cytosolic isoforms hCA I/II).Also, SLC-0111 has the ability to block human breast cancer invasion, delay tumor growth and diminish the cancer stem cell population in vivo [13][14][15][16].
Molecules 2018, 23, x 2 of 17 necessity to pay much effort to modify drug leads from the point of view of drug design to afford new bioactive chemical entities that offer improvements over current therapies.
In the current medical era, non-fused pyridines have emerged as a substantial class of heterocycles, which are endowed with diverse biological activities, chiefly anticancer activities [3][4][5].Several pyridine-based small molecules have been approved as anticancer drugs, to name just a few: Sorafenib I (Nexavar ® , Figure 1), Regorafenib II (Stivarga ® , Figure 1), Vismodegib III (Erivedge ® , Figure 1) and Crizotinib IV (Xalkori ® , Figure 1) [6][7][8].Subsequently, extensive efforts have been devoted to develop several pyridine-based derivatives as effective anticancer agents.BRN-103 V (Figure 1) is a nicotinamide derivative that suppresses the VEGF-induced phosphorylation of VEGFR-2 and the activation of AKT and eNOS.Also, it inhibits VEGF-induced migration, proliferation and capillary-like tube formation of HUVECs [9].SKLB610 VI (Figure 1) is a multi-targeted kinase inhibitor activity towards VEGFR-2 and FGFR-2.SKLB610 has anti-proliferative effects, especially on human colorectal cancer cell line HCT-116 and human NSCLC cell line A549, with significant activity on tumor xenografts in nude mice without evident toxicity [10].
On the other hand, urea derivatives represent one of the most useful classes of anticancer agents, with a wide range of activities towards various tumors [11].Urea functionality is the main pharmacophoric feature in several anticancer drugs such as Sorafenib I and Regorafenib II.Linifanib VII (Figure 1), an ureido indazole derivative, has selective inhibitory activity towards VEGFR and PDGFRs.Linifanib is in phase II clinical trials for patients with locally advanced or metastatic non-small cell lung cancer [12].SLC-0111 VIII (Figure 1), an ureido benzenesulfonamide derivative, is currently in phase I/II clinical trials as a potential anticancer drug.SLC-0111 exhibited selectivity towards inhibition of the transmembrane isoforms human carbonic anhydrase (hCA) IX/XII (over the cytosolic isoforms hCA I/II).Also, SLC-0111 has the ability to block human breast cancer invasion, delay tumor growth and diminish the cancer stem cell population in vivo [13][14][15][16].Recently, our research team has paid much attention to developing diverse novel small molecules, based on the pyridine core (structures IX and X, Figure 2) or incorporating the ureido functionality (structures XI and XII, Figure 2), as potent anticancer agents.These molecules displayed promising anticancer activities through different molecular and enzymatic targets, such as cytotoxic action [17,18], VEGFR-2 inhibition [19,20] or tumor-associated hCA isoform IX and XII inhibition [21].
Taking the above into account and as a continuation of our endeavor towards developing novel anticancer agents [22][23][24][25], it was thought worthwhile to extend our examinations to probe certain pyridine-urea derivatives displaying potent anticancer activity.In this study, a new series of pyridine-ureas 8a-n (Figure 2) was synthesized as potential anticancer agents.The latter synthesized Recently, our research team has paid much attention to developing diverse novel small molecules, based on the pyridine core (structures IX and X, Figure 2) or incorporating the ureido functionality (structures XI and XII, Figure 2), as potent anticancer agents.These molecules displayed promising anticancer activities through different molecular and enzymatic targets, such as cytotoxic action [17,18], VEGFR-2 inhibition [19,20] or tumor-associated hCA isoform IX and XII inhibition [21].
Taking the above into account and as a continuation of our endeavor towards developing novel anticancer agents [22][23][24][25], it was thought worthwhile to extend our examinations to probe certain pyridine-urea derivatives displaying potent anticancer activity.In this study, a new series of pyridine-ureas 8a-n (Figure 2) was synthesized as potential anticancer agents.The latter synthesized pyridine-ureas were evaluated for their growth inhibitory activity towards the proliferation of breast cancer MCF-7 cell line.Moreover, eight selected pyridines 8b, 8d, 8e, 8i, 8j and 8l-n were evaluated for their in vitro anticancer activity according to US-NCI protocol over 58 cancer cell lines.Moreover, the most active pyridines were screened in vitro for their inhibitory activity against VEGFR-2.Finally, several ADME descriptors were predicted for the active pyridines through a theoretical kinetic study.
Molecules 2018, 23, x 3 of 17 pyridine-ureas were evaluated for their growth inhibitory activity towards the proliferation of breast cancer MCF-7 cell line.Moreover, eight selected pyridines 8b, 8d, 8e, 8i, 8j and 8l-n were evaluated for their in vitro anticancer activity according to US-NCI protocol over 58 cancer cell lines.Moreover, the most active pyridines were screened in vitro for their inhibitory activity against VEGFR-2.Finally, several ADME descriptors were predicted for the active pyridines through a theoretical kinetic study.
Structures of some pyridine-/urea-based anticancer agents reported by our research group (IX-XII), and the target pyridine-ureas 8a-n.
Postulated structures of the newly synthesized pyridine-ureas 8a-n were in full agreement with their spectral and elemental analyses data (Supplementary Materials).

In Vitro Anti-Proliferative Activity
The in vitro anti-proliferative activity of the newly prepared pyridine-ureas 8a-n was examined against breast cancer MCF-7 cell line.The assay was carried out, as triplicates, utilizing the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay as described by T. Mosmann [26].Doxorubicin and Sorafenib were included in this assay as a reference drug.The results were expressed as median growth inhibitory concentration (IC 50 ) values that represent the compound concentrations required to afford a 50% inhibition of cell growth after 48 and 72 h of incubation, compared to the untreated controls (Table 1).Postulated structures of the newly synthesized pyridine-ureas 8a-n were in full agreement with their spectral and elemental analyses data (Supplementary Materials).

In Vitro Anti-Proliferative Activity
The in vitro anti-proliferative activity of the newly prepared pyridine-ureas 8a-n was examined against breast cancer MCF-7 cell line.The assay was carried out, as triplicates, utilizing the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay as described by T. Mosmann [26].Doxorubicin and Sorafenib were included in this assay as a reference drug.The results were expressed as median growth inhibitory concentration (IC50) values that represent the compound concentrations required to afford a 50% inhibition of cell growth after 48 and 72 h of incubation, compared to the untreated controls (Table 1).From the obtained results, it was obvious that most of the prepared pyridine-ureas 8a-n exhibited excellent to moderate anti-proliferative activity against MCF-7 breast cancer.Regarding the activity after 48 h treatment, compound 8e (IC 50 = 0.22 µM) was found to be the most potent derivative as it was 8.7 times more active than Doxorubicin (IC 50 = 1.93 µM) and 20 times more than Sorafenib (IC 50 = 4.50 µM).Also, compound 8n displayed excellent anti-proliferative activity against MCF-7 cells (IC 50 = 1.88 µM) which is comparable to Doxorubicin and better than Sorafenib.Furthermore, pyridines 8a-d, 8g, 8i, 8k and 8l displayed potent activity against MCF-7 cells with IC 50 range 3.03-7.03µM.Moreover, pyridines 8j and 8m were moderately active with IC 50 values of 10.09 and 23.02 µM, respectively.Unfortunately, compounds 8f and 8h did not display significant activity towards MCF-7 cells (IC 50 > 50 µM).
On the other hand, investigation of the anti-proliferative activity towards MCF-7 cells at 72 h of treatment elucidated that compounds 8g, 8j and 8l possessed higher activity at 72 h compared to 48 h.The drop in activity after longer incubation time, 72 h, could be explained by the rapid metabolism for such compounds or that the tested MCF-7 cells were resistant to compounds 8g, 8j and 8l upon 72 h treatment.With an exception of compound 8m (IC 50 = 13.1 µM) and compounds 8f and 8h (IC 50 > 50 µM), all the tested pyridines possessed potent activity against MCF-7 cells with IC 50 range 0.11-5.14µM, upon 72 h treatment.From the obtained results, it was obvious that most of the prepared pyridine-ureas 8a-n exhibited excellent to moderate anti-proliferative activity against MCF-7 breast cancer.Regarding the activity after 48 h treatment, compound 8e (IC50 = 0.22 μM) was found to be the most potent derivative as it was 8.7 times more active than Doxorubicin (IC50 = 1.93 μM) and 20 times more than Sorafenib (IC50 = 4.50 μM).Also, compound 8n displayed excellent anti-proliferative activity against MCF-7 cells (IC50 = 1.88 μM) which is comparable to Doxorubicin and better than Sorafenib.Furthermore, pyridines 8a-d, 8g, 8i, 8k and 8l displayed potent activity against MCF-7 cells with IC50 range 3.03-7.03μM.Moreover, pyridines 8j and 8m were moderately active with IC50 values of 10.09 and 23.02 μM, respectively.Unfortunately, compounds 8f and 8h did not display significant activity towards MCF-7 cells (IC50 > 50 μM).
On the other hand, investigation of the anti-proliferative activity towards MCF-7 cells at 72 h of treatment elucidated that compounds 8g, 8j and 8l possessed higher activity at 72 h compared to 48 h.The drop in activity after longer incubation time, 72 h, could be explained by the rapid metabolism for such compounds or that the tested MCF-7 cells were resistant to compounds 8g, 8j and 8l upon 72 h treatment.With an exception of compound 8m (IC50 = 13.1 μM) and compounds 8f and 8h (IC50 > 50 μM), all the tested pyridines possessed potent activity against MCF-7 cells with IC50 range 0.11-5.14μM, upon 72 h treatment.

NCI, USA Cytotoxicity Assay towards 60 Cancer Cell Lines
The structures of all the newly prepared pyridine-ureas 8a-n were submitted to the National Cancer Institute (NCI) Developmental Therapeutic Program.Eight compounds 8b, 8d, 8e, 8i, 8j and

NCI, USA Cytotoxicity Assay towards 60 Cancer Cell Lines
The structures of all the newly prepared pyridine-ureas 8a-n were submitted to the National Cancer Institute (NCI) Developmental Therapeutic Program.Eight compounds 8b, 8d, 8e, 8i, 8j and 8l-n were selected to be examined for their in vitro growth inhibitory activity.The anticancer assays were carried out according to the protocol of the Drug Evaluation Branch, NCI, Bethesda [27][28][29].The selected pyridine-ureas were tested at one dose primary anticancer assay against a panel of approximately 58 cancer lines (concentration 10 −5 M).The tested human cancer cell lines emerged from nine different cancer subtypes: leukemia, colon, melanoma, ovarian, lung, CNS, renal, breast and prostate cancers.The sulforhodamine B (SRB) protein assay was adopted to estimate the cell viability and growth [30].The results were reported as mean-graph of the percentage growth of the treated cells, and presented as percentage growth inhibition (GI%) caused by the test compounds (Table 2).The gained data disclosed that most tested pyridines possessed distinctive patterns of selectivity and sensitivity towards the different NCI cancer cell panels (Supplementary Material).

Cytotoxic Activity against Non-Tumorigenic Human WI-38 Cells
The cytotoxic activity of compounds 8b and 8e were evaluated against non-tumorigenic human lung fibroblast WI-38 cell line utilizing the MTT colorimetric assay developed by T. Mosmann [26] (Table 3).Interestingly, both the tested compounds 8b and 8e displayed non-significant weak cytotoxic activity towards WI-38 cell line with IC50 values of 91.5 ± 4.53 and 83.14 ± 5.21 μM, respectively.

Cytotoxic Activity against Non-Tumorigenic Human WI-38 Cells
The cytotoxic activity of compounds 8b and 8e were evaluated against non-tumorigenic human lung fibroblast WI-38 cell line utilizing the MTT colorimetric assay developed by T. Mosmann [26] (Table 3).Interestingly, both the tested compounds 8b and 8e displayed non-significant weak cytotoxic activity towards WI-38 cell line with IC 50 values of 91.5 ± 4.53 and 83.14 ± 5.21 µM, respectively.

VEGFR-2 Inhibitory Assay
In an attempt to gain further mechanistic insights for the promising anti-proliferative activity of the prepared pyridine-ureas, compounds 8b and 8e, with significant anti-proliferative activity, were evaluated in vitro for their VEGFR-2 inhibitory activity.Sorafenib, a pyridine urea-based and well-known FDA-approved VEGFR-2 inhibitor, was used as the reference drug.The results are reported as median inhibition concentrations (IC 50 ) which are determined as triplicate determinations from the standard curve and listed in Table 4. Results revealed that the tested pyridines 8b and 8e exhibited modest VEGFR-2 inhibitory activity with IC 50 values of 5.0 ± 1.91 and 3.93 ± 0.73 µM, respectively, with respect to the reference drug Sorafenib (IC 50 = 0.09 ± 0.01 µM).

Physicochemical Properties and ADME Profiling
With a view to investigate drug-like physicochemical and pharmacokinetics properties of the target pyridine-ureas, several ADME descriptors for the active pyridines (8a, 8b, 8e, 8g, 8i, 8l and 8n) were estimated, in addition to the assessment of the criteria of both the Lipinski rule of five [31] and Veber rule [32].A computer-aided theoretical kinetic study was performed via Discovery Studio 2.5 software (Accelrys, San Diego, CA, USA) to estimate different ADME descriptors for the most active anti-proliferative pyridines 8a, 8b, 8e, 8g, 8i, 8l and 8n, Table 5.
All the tested pyridines exhibited low aqueous solubility levels along with a good level of human intestinal absorption.Regarding blood-brain barrier penetration, the tested pyridines were discovered to possess certain penetrability to the blood-brain barrier.On the other hand, the investigated pyridines passed the filter of both the Lipinski rule of five and Veber rule, which implies that the synthesized pyridines have the privilege to possess good oral bioavailability, Table 6.81.71 1 The upper limit of the number of hydrogen bond donors is 5; 2 the upper limit of the number of hydrogen bond acceptors is 10; 3 the upper limit of the molecular weight is 500; 4 the upper limit of the AlogP (the log value of octanol-water partition coefficient) is 500; 5 the upper limit of the number of rotatable bonds is 10; 6 the upper limit of the polar surface area is 140 Å 2 ; * the upper limit of the sum of the hydrogen bond donors and acceptors is 12.

Chemistry
Melting points were measured with a Stuart melting point apparatus (Bibby Scientific Limited, Staffordshire, UK) and were uncorrected.Infrared measurements (neat, thin film) were carried out using Schimadzu FT-IR 8400S spectrophotometer (Shimadzu, Kyoto, Japan). 1 H-NMR and 13 C-NMR experiments were carried out using Bruker AVF-400 (400/100 MHz) and AVC-500 (500/125 MHz) (Bruker, Karlsruhe, Germany), respectively.Chemical shifts (δ H ) are reported relative to TMS as the internal standard.All coupling constant (J) values are given in hertz.Chemical shifts (δ C ) were reported as follows: s, singlet; d, doublet; m, multiplet.High-resolution mass spectra (EI and ESI) were recorded using a Bruker MicroTOF spectrometer (Bruker Daltonics, Bremen, Germany) by the internal service at the University of Oxford.Analytical thin layer chromatography (TLC) (Merck KGaA, Darmstadt, Germany) on silica gel plates containing UV indicator was employed routinely to follow the course of reactions and to check the purity of products.All reagents and solvents were purified and dried by standard techniques.Compounds 2a,b, 3a,b, and 4a,b [17,33] were previously prepared.To a suspension of esters 3a,b (10 mmol) in methyl alcohol, 99% hydrazine hydrate (3 mL) was added.The reaction mixture was heated under reflux for 2 h.The precipitate formed was collected by filtration while hot, washed with water, dried and recrystallized from ethanol to produce hydrazides 4a,b in 80% and 86% yield, respectively.The physical properties and spectral data of 4a,b were identical with those reported [28].In an ice bath, a mixture of hydrazides 4a,b (5 mmol) and sodium nitrite (0.5 g, 7 mmol) was stirred in glacial acetic acid for 1 h, then stirring was continued at room temperature for another 1 h.The obtained solid was collected by filtration, washed with cold water and air-dried to furnish 6-(4-methoxyphenyl/3,4-dimethoxyphenyl)-2-methylnicotinoyl azide 5a,b, which used in the next reaction without further purification.Then, the appropriate nicotinoyl azide 5a,b was heated under reflux in dry xylene for 1 h before addition of anilines 7a-g.The reaction mixture was refluxed for 3 h then allowed to cool to room temperature.The formed precipitate was filtered off, washed with cold acetone, dried and recrystallized from dioxane to furnish the target pyridines 8a-n.[26,34,35].The 50% inhibitory concentration (IC 50 ) was estimated, after 48 and 72 h for MCF-7 cells, from graphic plots of the dose response curve for each conc.using Graphpad Prism software (GraphPad Software, Inc., San Diego, CA, USA).The data presented are the mean of at least three separate experiments.

In Vitro Cytotoxic Activity by NCI-USA
The anticancer assays were performed in accordance with the protocol of the Drug Evaluation Branch, NCI, Bethesda [27][28][29].A 48 h drug exposure protocol was used and sulforhodamine B (SRB) protein assay [30] was applied to estimate the cell viability and growth, as reported earlier [36,37].

Measurement of Inhibitory Activity against VEGFR-2
VEGFR-2 enzyme inhibition was measured for pyridines 8b and 8e using an anti-phosphotyrosine antibody with the Alpha Screen system (PerkinElmer, Waltham, MA, USA) according to the manufacturer's protocol and referring to reported instructions [19].The concentration of the test compound causing 50% inhibition (IC 50 ) was calculated from the concentration-inhibition response curve (triplicate determinations) and the data were compared with sorafenib as standard VEGFR-2 inhibitor.

Physicochemical Properties and ADME Profiling
Physicochemical properties and ADME profiling for the active pyridines (8a, 8b, 8e, 8g, 8i, 8l and 8n) were performed using Discovery Studio 2.5 (Accelrys, San Diego, CA, USA).The examined pyridines were drawn as a small library and prepared via prepare ligand protocol in order to find the suitable orientation in 3D.Then, the prepared library was filtered adopting the Lipinski and Veber rules protocols.ADME profiling was predicted for the designed library using ADME descriptors protocol.

Conclusions
In summary, herein we report the synthesis of a novel series of pyridine-ureas 8a-n.All the prepared pyridines were evaluated for their anti-proliferative activity towards the breast cancer MCF-7cell line.Pyridines 8e and 8n were the most active members towards MCF-7 cells (IC 50 = 0.22 and 1.88 µM after 48 h treatment; 0.11 and 0.80 µM after 72 h treatment, respectively).Furthermore, eight selected pyridines 8b, 8d, 8e, 8i, 8j and 8l-n were examined for their in vitro anticancer activity according to US-NCI protocol.Pyridines 8b and 8e emerged as the most effective anticancer agents in the NCI assay with mean inhibition = 43 and 49%, respectively.Both 8b and 8e exhibited anti-proliferative activity against all tested cancer cell lines from all subpanels (GI for 8b; 12-78%, GI for 8e; 15-91%).Pyridines 8b and 8e were screened in vitro for their inhibitory activity against VEGFR-2.Both compounds inhibited VEGFR-2 at micromolar IC 50 values 5.0 ± 1.91 and 3.93 ± 0.73 µM, respectively.The most active pyridines were filtered according to the Lipinski and Veber rules and all of them passed these filters.Finally, several ADME descriptors were predicted for the active pyridines through a theoretical kinetic study.Further mechanistic studies are in progress in our laboratories and will be reported upon in the future Supplementary Materials: The following are available online.Spectral data and NCI One Dose Mean Graph.

Figure 3 .
Figure 3. Mean % growth inhibitions of the tested pyridines over NCI 58 cell line panel.

Figure 3 .
Figure 3. Mean % growth inhibitions of the tested pyridines over NCI 58 cell line panel.

Figure 4 .
Figure 4. (A) The most sensitive cell lines towards the target pyridines 8b, 8d, 8e and 8i according to the GI%; (B) the most sensitive cell lines towards the target pyridines 8j, 8l, 8m and 8n according to the GI%.

Figure 4 .
Figure 4. (A)The most sensitive cell lines towards the target pyridines 8b, 8d, 8e and 8i according to the GI%; (B) the most sensitive cell lines towards the target pyridines 8j, 8l, 8m and 8n according to the GI%.

2 .
General Procedure for the Preparation of Target Pyridine-Ureas 8a-n

Author
Contributions: H.A.A.-A.and W.M.E.conceived and designed the experiments; W.M.E., M.E.-N.and H.S.I. carried out the experiments; H.A. and H.S.I. analyzed and interpreted the data; H.A.A.-A., W.M.E. and M.E.-N.prepared the manuscript; All authors have read and approved the final manuscript.Funding: "This research received no external funding" Conflicts of Interest: The authors declare no conflict of interest.

Table 1 .
In vitro anti-proliferative activity of pyridine-ureas 8a-n against MCF-7 breast cancer cell line.

Table 1 .
In vitro anti-proliferative activity of pyridine-ureas 8a-n against MCF-7 breast cancer cell line.

Table 2 .
Percentage growth inhibition (GI%) of in vitro subpanel tumor cell lines at 10 µM concentration for compounds 8b

Table 3 .
Cytotoxic activity of compounds 8b and 8e against non-tumorigenic human lung fibroblast WI-38 cell line.IC 50 values are the mean ± S.D. of three separate experiments. a
a IC 50 values are the mean ± S.D. of three separate experiments.

Table 5 .
ADME studies results for all the synthesized compounds.The base 10 logarithm of the molar solubility as predicted by the regression model based by DS; 2 Categorical solubility level.

Table 6 .
Lipinski rule of five parameters results for all the synthesized compounds.