And to determine the anti-inflammatory and anti- cancer mechanisms of the hybrid substances, studies of PGE2 production inhibition activity, cell cycle analysis meth[r]
(1)AND TRAINING SCIENCE AND TECHNOLOGY GRADUATE UNIVERSITY OF SCIENCE AND TECHNOLOGY
-
NGUYEN THI THUY HANG
“SYNTHESIS AND ANTIINFLAMMATORY, ANTIPROLIFERATIVE ACTIVITIES OF NEW COXIB–COMBRETASTATIN HYBRIDS’’
Scientific Field: Organic Chemistry Classification Code: 9.44.01.14
DISSERTATION SUMMARY
(2)
Vietnam Academy of Science and Technology Scientific Supervisors:
1 Assoc Prof Dr Ngo Quoc Anh Assoc Prof Dr Vu Dinh Hoang
1st Reviewer: 2nd Reviewer: 3rd Reviewer:
The dissertation will be defended at Graduate University of Science And Technology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Cau Giay District, Ha Noi City
At … hour… date… month … 2021
(3)INSTRODUCTION
1. The urgency of the thesis
Cancer is a group of diseases that involve disorganized cell division and cells that have the ability to invade other tissues by either growing directly into nearby tissue or moving to multiple locations different (metastatic) According to the Global Cancer Organization GLOBOCAN 2018, there are currently more than 300,000 people living with cancer nationwide, there are 164,671 new cases, 114,871 people die from this disease Globally, there are about 23 million people infected, of which more than 14 million people are newly infected and million hundred thousand people die The World Health Organization (WHO) ranked Vietnam in the top 50 countries in the top of the cancer map
Researching to find cancer treatment drugs with few side effects is one of the directions that is always interested in the scientific community Among current treatments, chemotherapy is a cancer treatment that uses one or more anticancer drugs - cytotoxic One of the anti-cancer drugs, used today in chemotherapy, influences the cell cycle to inhibit cancer cell growth and subsequently induce apoptosis (apoptosis)
(4)practice, it is still necessary to look for new agents that can overcome the limitations of resistance and the undesirable side effects of these therapies current method [3]
2 Objectives of the dissertation
1 Structured design of coxib - combretastatin hybrid compounds Synthesize coxib - combretastatin hybrid compounds
3 Screening for anti-cancer and anti-inflammatory activities of hybrid compounds
4 Identifying anti-inflammatory and anti-cancer mechanisms of hybrid compounds
5 Docking of a hybrid coxib - combretastatin compound with two purpose effect COX2 and tubulin
3 The main research contents of the thesis
- Research on coxib - combretastatin hybrid compounds synthesis
- Determination of the structure of coxib-combretastatin hybrid compounds - Screening the activity of coxib - combretastatin hybrid substances
- Study on mechanism of action of some coxib - combretastatin hybrid substances
(5)(6)DISSERTATION CONTENTS CHAPTER LITERATURE REVIEW
General presentation of anticancer compounds by tubulin inhibitory mechanism, profile of tubulin mechanism The group of tubulin-based compounds has always been a topic of concern in the field of anti-cancer drug research Combretastatin compounds with rich biological activity have been used in the treatment of a number of cancers They are known for their cytotoxic activity by inhibiting tubulin polymerization at the colchicine site [ 2] Up to now, these cancer treatment compounds with this mechanism are still being widely used and always a research direction that receives a lot of attention
Overview of combretastatin compounds, which belong to the class of cis -stilbene, a rich source of pathogens in the search for new drugs, typical compounds such as resveratrol and combretastatin A-4 phosphate are currently is clinically tested to treat Alzheimer's disease and cancer The recently isolated stilbene has been shown to have a diverse range of biological activities, including antioxidant, antibacterial, anti-malarial, cytotoxic, liver protective and anti-inflammatory properties Combretastatin A-4 (CA4) is also considered to be a potential cytotoxic agent by strongly inhibiting microtubule polymerization by binding to the binding point of colchicine on tubulin CA-4 is highly toxic on many cancer cell models, making it a very interesting target structure
(7)substance known for its potent anti-inflammatory activity, which selectively inhibits COX2 through its action Prostaglandins induce inflammation and pain without effects on prostaglandins COX1 that have a protective effect on the gastrointestinal tract Furthermore, Celecoxib inhibits the proliferation of human breast cancer in vitro models such as MCF7 and MDAMB-231 Some studies indicate that celecoxib and related compounds can induce cell cycle arrest at G0 /G1 stage leading to apotosis cyclic apoptosis, inhibition of tumor growth and prevents tumor angiogenesis in the absence of COX2
(8)CHAPTER EXPERIMENTS 2.1 Materials and equipments
2.1.1 Materials 2.1.2 Equipments
2.2 Methods
2.2.1 Organic synthesis method 2.2.3 Biological activity test method
2.3 Synthesis of coxib- combrestatin hybrid compounds
2.3.1 Synthesis of ester derivatives of coxib - combretastatin hybrid ester hybrid
Figure 2.2 Ester derivative synthesis of coxib - combrestatin hybrids compound
(i) Alkaline: t-BuOLi (3 mmol, eq), refluxe, (ii) Ethyl chlorooxoacetate (1 mmol, eq) (77), ml THF; (iii) HCl (4 mmol); refluxe, ml C2H5OH dry;
phenylhydrazin (1 mmol, eq) (78)
Synthesized 20 hybrids esters of coxib - combretastatin ester form substances from 79 to 98
2.3.2 Synthesis of coxib - combrestastatin hybrids compounds containing groups CF3
(9)(a) 100 (1.0 mmol), 99 etyl trifluoroacetate (1,2 eq) and NaH (2,5 eq) in THF (5 mL), h
(b), EtOH (5 mL), axit (1.0 eq); arylhydrazin hydrochloride 78 (1.0 mmol) is added consecutively to the residue and restored for hours Substance 102 was isolated by column chromatography
2.3.3 Synthesis of acid derivatives of coxib - combretastatin hybrid
Figure 2.8 Synthesis of coxib - combretastatin hybrids
(i) Alkaline: t-BuOLi (3 mmol, eq), refluxe, (ii) Ethyl chlorooxoacetate (1 mmol, eq) (77), ml THF; (iii) HCl (4 mmol); refluxe, ml C2H5OH dry;
phenylhydrazin (1 mmol, eq) (78) The product obtained after isolation through column chromatography was dissolved in the solvent system THF / MeOH / H2O = 3: 1: 1, then NaH (1,2 eq) was added to the mixture Carry out the reaction in hours to obtain compounds acid hybrids 103-122
successful synthesis of 20 acid hybridization of coxib – combretastatin hybrids 103-122
2.5 Biologically active testing of research compounds
(10)CHAPTER RESULTS AND DISCUSSIONS
The advantages of using a hybrid molecule over co-combination of multiple drugs at the same time may improve the limitations of adverse effects and resistance [107] Despite its outstanding activity, combretastatin still has many undesirable effects This is why the team aims to combine combretastatin, an anticancer compound, and celecoxib, a COX2-engineered anti-inflammatory agent, as derivatives for new hybrid compounds in hopes of finding new It has interesting biologically active properties such as the anticancer and anti-inflammatory properties of the parent substance and has less side effects
3.1 Design of the structure and biological activity of the hybrids
3.1.1 Design of hybrid molecular structure
(11)Figure 3.1. C Structure of celecoxib, combretastatin A4 and the coxib– combretastatin hybrids
3.1.2 Design of hybrid molecular biology activities
(12)multi-target drug by combining COX-2-selective inhibition with nitric oxide (NO)-dependent activities has been initiated [112-114]
In infammation, the activated immune cells such as macrophages secrete everal infammatory mediators such as proinfammatory cytokines, nitric oxide (NO) and prostaglandin E2 (PGE2) In our previous study, tested compounds and celecoxib were examined for their efects on nitric oxide (NO) production in LPS-activated murine macrophage RAW 264.7 cells 264.7 cells [115] Prostaglandin E2 is present in high concentrations in breast tumors and in metastatic cancers, in the absence of estrogen and progesterone receptors [116-117] Hence, the ability to inhibit production of PGE2 can be considered a good strategy in the design of anticancer agents However, the MCF7 breast cancer cells produced very low levels of PGE2, according to another report [109] Therefore, the active efficacy of the hybrid compounds was tested for the inflammatory response induced by LPS instead of by inhibiting PGE2 on MCF7 [118]
Therefore, cancer cell lines HT-29, Hep-G2, MCF-7 and inhibition of NO production were selected as initial tools to screen for anti-inflammatory activity of the study agent class And to determine the inflammatory and anti-cancer mechanisms of the hybrid substances, studies of PGE2 production inhibition activity, cell cycle analysis methods, apoptosis methods by nuclear staining cells with Hoechst 33342, study of apotosis-inducing activity by caspase-3 indicator, study of apoptosis induction by FITC-anexin V and PI stained cell photometric method Finally, substances selected and studied on biological mechanisms will be tested to interact with tubulin and COX2 targets by molecular docking method to confirm the biologic accuracy of the model in vitro
3.2 Synthesis of coxib - combretastatin hybrids
(13)oxalyl chloride arylhydrazine hydrochloride derivatives (Figure 2.1 , Figure 2.2, Figure 2.7, Figure 2.8) [109] Reaction of Claisen condensation between two components 1,2-diarylethanon and ethyl oxalyl chloride with alkaline agent using tert-BuOLi, obtained the lithium salt intermediate product of ethyl 2,4-dioxo-3,4-diarylbutanoate , the reaction is continued with arylhydrazine hydrochloride through the Knorr reaction catalyzed by hydrochloric acid to produce triarylpyrazole-3-carboxylate (hybrids compound 79 to 98) and (hybrids compound 103 to 122) A cycle of celecoxib derivatization was also applied (Figure 2.4, Figure 2.5) 3,4,5-trimethoxyphenyl - 1,1,1-trifluoro-2,4-butanedion (65, 102) obtained from Claisen condensation between acetophenone and ethyl trifluoroacetate derivatives then reacts with 4-sulfamidophenyl hydrazine halide salt to form 3,4,5-trimethoxyphenyl, a substance with structure similar to celecoxib
According to the reaction procedures described in Figure 2.1, Figure 2.2, 20 coxib-combretastatin hybrid compounds were successfully synthesized with efficiency from 64 to 83% (Table 3.1) 02 compounds containing CF3 group with difference rates are 95% respectively 78% (table 3.2) 20 acidic compounds of coxib - combretastatin hybrid with efficiency from 54 - 74% (Table 3.3); The substances are structurally determined by the methods of purification and nuclear magnetic resonance spectroscopy method NMR 1D, 2D
Of the hybrid compounds obtained, substances 79 to 98 and 102 to 122 were synthesized for the first time and never previously published And 01 compound has been published and also used as a comparator is celecoxib (65) which is known for many studies [78-79]
(14)3.3.1 Screening for biological activity of coxib - combretastatin hybrids
Screening activity of 20 ester-based coxib-combretastatin hybrid compounds and 02 hybrid compounds containing groups CF3
(15)inhibitory activity, emphasizing that they may have a better safety in the cardiovascular system than celecoxib [110]
Table 3.4 The cytotoxic effect of ester-type hybrid compounds on three cancer cell lines HT-29, Hep-G2, MCF7 and inhibition of NO production
Entry Comp R R’ R’’ IC50 (µM) Inhibits NO production
HT-29 Hep-G2 MCF-7
1 102
3,4,5-triOMe
4-NH2SO2 1.95±0.11 43.0 ± 6.0 39.96± 3.36 4.63± 0.59
2 96 H 4-F 4-NH2SO2 81.6 ± 10.6 18.7 ± 1.7 29.37± 1.90 7.97±0.51
3 81 H H 4-CN 116.83±7.7 37.6±3.5 >254 8.20± 0.93
4 82 H H 4-NO2 129.88±15.7 21.2±2.1 >242 9.67± 0.92
5 94 H 4-OH 4-NH2SO2 92.3 ± 7.2 18.8 ± 1.5 14.51±1.36 8.90± 0.79
6 93 H 4-Br 4-NH2SO2 73.7 ± 2.6 23.5 ± 1.8 35.48±2.85 19.27±0.27
7 91 H 4-Br 4-CN 116.01±13.6 32.6± 3.0 >211 20.16± 1.83
8 92 H 4-Br 4-NO2 91.58±7.8 153.2± 10 >203 21.30± 1.40
9 89 H 4-Br 4-OMe 21.01±4.3 39.6±2.9 >209 21.51± 1.48
10 90 H 4-Br 4-CF3 > 233 10.0±0.7 75.87±4.41 21.65± 1.86
11 97 H 4-Cl 4-NH2SO2 122.2 ± 9.7 14.1 ± 37.04±4.95 23.79± 3.11
12 85
4-OMe
4-OMe 4-CF3 134.10±15.7 107± 10 >201 29.80± 2.71
13 43 4-Me 4-NH2SO2 83.59 ± 5.0 58.4 ± 3.4 37.11±2.14 30.67±3.79
14 98 H 4,6-
diOMe
4-NH2SO2 4.41±0.17 136.0 ± 12 >197 39.93± 2.64
(16)Table 3.5 Comparison of cytotoxic effects of acid and ester hybrid compounds on MCF-7 cancer cells and inhibition of NO production
16 79 H H 4-OMe 30.0±13.4 >251 21.43±1.93 43.19±4.86
17 87
4-OMe
4-OMe 4-NO2 148.85±8.4 >211 >211 48.81± 2.62
18 86
4-OMe
4-OMe 4-CN 76.42±9.4 110.3±7.0 >220 48.86± 3.13
19 84
4-OMe
4-OMe 4-OMe 67.06±3.7 160±15 >218 49.32± 1.72
20 80 H H 4-CF3 59.52±5.5 17.3±2.0 63.17± 3.01 50.66± 4.46
21 88
4-OMe
4-OMe 4-NH2SO2 103.4 ±10.5 99.0±6.3 58.57± 4.70 68.43± 7.05
22 98 H 4,6-diOH 4-NH2SO2 > 208.5 173.9 ± 12 64.03± 5.75 83.60± 3.44
L-NMMA 8.39±0.31
Ellipticine 0.18± 0.04 0.44± 0.05 0.36± 0.04
STT R R’ R’’
IC50 (µM) Este (NO) Acid (NO) Este
(MCF-7)
Acid (MCF7)
1 3,4,5-triOMe 4-NH2SO2 1.95±0.11 43.0 ± 6.0 39.96± 3.36 4.63± 0.59
2 H 4-F 4-NH2SO2 81.6 ± 10.6 194.5±7.6 7.97±0.51 187.7±9.5
3 H H 4-CN 116.83±7.7 256.7±22.1 8.20± 0.93 154.5±15.1
4 H H 4-NO2 129.88±15.7 >261.1 9.67± 0.92 187.6±10.4
5 H 4-OH 4-NH2SO2 92.3 ± 7.2 75.8±2.6 8.90± 0.79 34.6±2.6
6 H 4-Br 4-NH2SO2 73.7 ± 2.6 >202.0 19.27±0.27 149.5±7.3
(17)Note: *: Statistics column of ester activity **: Statistics column of acidic activity
The super-active hybrid compounds 102, 96, 81, 82, 94 and celecoxib continued to be transferred to the PGE2 production test to further screen for the anti-inflammatory and anticancer activities of the active ingredient
3.3.2 Study on anti-inflammatory and anti-cancer mechanisms
8 H 4-Br 4-NO2 91.58±7.8 111.2±8.7 21.30± 1.40 34.0±6.4
9 H 4-Br 4-OMe 21.01±4.3 >223.7 21.51± 1.48 >223.7
10 H 4-Br 4-CF3 > 233 80.0±6.3 21.65± 1.86 129.5±9.7
11 H 4-Cl 4-NH2SO2 122.2 ± 9.7 >220.7 23.79± 3.11 191.1±20.6
12 4-OMe 4-OMe 4-CF3 134.10±15.7 140.3±16.7 29.80± 2.71 136.0±8.4
13 4-Me 4-NH2SO2 83.59 ± 5.0 30.67±3.79 30.0±3.3
14 H 4,6- diOMe 4-NH2SO2 4.41±0.17 >208.7 39.93± 2.64 94.0±6.4
15 H H 4-NH2SO2 > 233 >239.8 40.54±2.56 178.9±12.9
16 H H 4-OMe 30.0±13.4 >271.7 43.19±4.86 209.3 ± 20.0
17 4-OMe 4-OMe 4-NO2 148.85±8.4 185.9±9.8 48.81± 2.62 31.8±3.7
18 4-OMe 4-OMe 4-CN 76.42±9.4 157.0±12.2 48.86± 3.13 83.0±10.3
19 4-OMe 4-OMe 4-OMe 67.06±3.7 145.7±15.2 49.32± 1.72 75.7±6.9
20 H H 4-CF3 59.52±5.5 109.4±5.8 50.66± 4.46 203.8 ± 5.1
21 4-OMe 4-OMe 4-NH2SO2 103.4 ±10.5 >209 68.43± 7.05 150.0±14.7
22 H 4,6-diOH 4-NH2SO2 > 208.5 >222.0 83.60± 3.44 >222.0
L-NMMA 8.39±0.31
(18)3.3.2.1 Study on PGE2 production inhibition activity
Typical compounds 102, 96, 81, 82, 94, celecoxib continued to be tested for the ability to inhibit PGE2 production
Table 3.7 Effect of test compounds on PEG2 production in LPS-stimulated RAW 264.7 macrophages
PGE2 (pg/ml) Concentration
(µM) 102 96 81 82 94
Concentration
(µM) Celecoxib (43)
20 65.31 223.82 253.60 155.07 246.88 100 41.45
4 133.59 326.20 277.23 383.49 488.82 20 129.83
0.8 225.56 399.56 490.73 478.52 560.34 278.64
LPS 329.60
3.3.2.2 Cell cycle analysis
Compounds 96, 81, 82 and 94 increased the number of cells in G0/G1 phase and decreased them in S and G2/M phases as celecoxib did Celecoxib and related compounds ere also known to be able to inhibit
tumor growth and to induce apoptosis Some research also resulted in the stable G0 /G1 block efect of celecoxib with unclear involvement of COX-2 as well as of PGE2 [104] On other hand, compound 102 caused G2/M phase arrest as evidenced by the increase in cell number with a concomitant decrease in cells in phase S G2/M phase arrest prevents cells from exiting mitosis, a feature shared by microtubule inhibiting agents such as colchicine or combretastatins [78] In case of compound 102, replacement of the double bond of CA4 with pyrazole ring of celecoxib and maintaining trimethoxybenzene moiety of CA-4 were demonstrated to be crucial to obtain relevant cytotoxic and antimitotic efects [98, 115] Table 3.8 Percentage of cell by phases of tested compounds in MCF7cells entrie
Entries Compound Percentage of cell by phases (%) % G0/G1 % S % G2/M
1 Negative control
(DMSO 0,5%) 42.46 40.47 13.15
2 102 (10 µM) 40.72 35.09 21.04
3 96 (10 µM) 49.00 34.17 14.17
4 81 (10 µM) 45.22 34.46 17.45
5 82 (10 µM) 50.96 30.75 14.80
6 94 (10 µM) 49.55 34.50 14.34
(19)(-) control 102 96
81 82 94
Celecoxib (43)
Figure 3.3 Cell cycle analysis of tested compounds including 102, 96, 81, 82, 94 and celecoxib at the concentration of 10 µM on MCF-7 human breast cancer cells using Novocyte fow cytometry system (the experiment has been done one time)
3.3.2.3 Research results of apoptosis-causing activity of compounds 82 and 102
Since compounds 102 and 82 are potential inhibitors of antiproliferative activity, inhibiting PGE2 production more potent than celecoxib and selectively inhibiting cell cycle, it was selected for further evaluation of potential for apoptosis
(20)Table 9. Percent of condensation or fragmentation in the cell nucleus caused by compounds 102 and 82
Tế bào Apoptosis (%)
82 (20 µM) 82 (10 µM) 82 (5 µM) 102 (20 µM) 102 (10 µM) 102 (5 µM) Camptotheci n (5µM) (-) Control
5.33 ± 0.34 2.69± 0.22
2.42±
0.27 12.38± 0.96
5.18± 0.33
3.55±
0.39 22.71± 2.01
2.71± 0.29
The study samples obtained MCF7 cell image after being stained with Hoechst 33342 at different concentrations:
Negative control Camptothecin (5 µM)
102 (20 µM)
102 (10 µM) 82 (20 µM) 82 (10 µM)
Figure 3.4. Image of MCF7 cells under the influence of research samples stained with Hoechst 33342 at different concentrations
Study of apotosis-inducing activity by caspase-3 indicator
Table 3.10. Activation of caspase by compounds 102 and 82
% Tế bào Apoptosis Comp 102
(20 µM)
102 (10 µM) 102 (5 µM) 82 (20µM) 82 (10 µM) 82 (5 µM) Camptothecin (5µM) (-) control
Mean 1.36* 1.21* 1.01 1.08 1.12 1.11 1.67** 1.00
(SD) 0.033 0.013 0.037 0.023 0.056 0.040 0.020 0.070
(21)Study on apoptosis induction by FITC-anexin V and PI stained cell photometric method
Table 3.11. Percentage of apoptosis cells
Samples % Necrotic
cells
% Early
apoptosis cell
% % late
apoptosis cell
%
Apoptosis cells
Negative control 0.22 18.32 1.90 20.22
82 (20 µM) 1.39 18.49 4.08 22.57
82 (10 µM) 0.27 21.62 4.59 26.21
82 (5 µM) 0.16 22.74 3.27 26.01
102 (20 µM) 0.42 42.24 6.73 48.97*
102 (10 µM) 0.49 19.90 3.45 23.35
102 (5 µM) 0.27 15.84 2.17 18.01
Camptothecin (5µM) 0.08 73.83 4.96 78.79**
Ghi chú: * P<0.05; ** P<0.01
Cell incubated with compound 82 (20 µg/ml) in 48h
Cell incubated with compound 82 (10 µg/ml) in 48h
Cell incubated with compound 82 (5 µg/ml) in 48h
Cell incubated with compound 102 (20 µg/ml) in 48h
Cell incubated with compound 102 (10 µg/ml) in 48h
(22)Cell incubated with 0.5% DMSO 48h
Cell incubated with5 µM camptothecin in 48 h
Fig 3.5. Cell apoptosis of tested compounds via Dead Cell Apoptosis Kit with Annexin V FITC and PI for fowcytometry The x-axis represents the level of FITC-Annexin V staining; the y-axis represents the level of PI staining in log units
The conclusions determine the apoptosis induction potential of two study samples
Through the determination of apoptosis induction through three methods of detecting apoptosis of the active substance by staining the cell nucleus with Hoechst 33342, caspase - and cytometry can be summarized:
Sample 82 at test concentrations slightly increased the proportion of cells in early apoptosis and late apoptosis compared with negative control, reaching 22.57-26.21%
Sample 102, at a concentration of 20 µg / ml increased the incidence of early and late apoptosis cells from 18.3% to 42.2% and 1.90% to 6.7% in the negative control, respectively At lower concentrations there was a slight increase (10 µg / ml and /g / ml) in the incidence of apoptosis compared to the negative control
Camptothecin - positive control increased apoptosis rate, reaching from 78.79%
Sample 82 has not clearly shown the ability to induce apoptosis to research concentrations in the trials performed;
(23) Sample 102 at a concentration of 20 M induces apoptosis on MCF7 cells through: inducing concentration or fragmentation of the cell nucleus at a rate of 12,386%; induction of caspase -3 generation with concentration increased 1.36 times compared with negative control (P <0.01); Apoptosis increased and especially early apoptosis increased 42.24% through cell photometric technique (P <0.05)
Thereby, based on the results of cytotoxic activity, NO, PGE2 anti-inflammatory and apoptosis induction, we confirmed the mechanism and activity characteristics of the obtained hybrid compounds The results showed that the hybrid molecule in terms of both molecular structure and biological activity according to the design target model of the original hybrid molecule We have continued to opt for molecular docking analysis to simulate the interaction between engineered hybrid with COX2 and tubulin targets
4 Molecular docking study
According to AutoDock's rating criteria 4.2.6, the negative the value of energy, the better the ability of the compound to bind to the targeted receptor Compounds 102 and 82 were shown to be two potential candidates with good binding energies for both targets and can be considered as potential COX2 / tubulin dual inhibitors, their interactions analyzed added using Model Studio Visualizer The resulting computation can be used to better predict a protein mode inhi itor ased on the differen e Δ etween the COX2 intera tion value and tubulin (Equation 1–2)
ΔG=Do k s oreCOX2−Dockscroretubulin (1)
ΔLE=LECOX2−Letubulin (2)
Dock pose of compound 102 in the active site of COX2 showed three hydrogen bonds interaction between the sulfonamide group (SO2NH2) and
(24)residues Arg106 of COX2 and Ala250 of tubulin (distance = 2.99; 3.18 Å, respectively); particularly, the NO2 group of 6cwas observed to interact only
with residue Phe504 of COX2 This fnding supports our assumption that the sulfonamide and ethyl ester groups play an important role for the dual activities of compounds 102 and 82 On the other hand, the trimethoxybenzene and other aryl substituents are assumed to characterize the specifc binding of each compound to protein targets
Table 3.12. Set of designed compounds with respective docking score on COX2 and tubulin model
Designed compounds Dock score (kcal/mol)
COX2 (3LN1) Tubulin (1Z2B)
102 -11.90 -8.06
82 -11.92 -9.37
Combretastatin A4 -9.63 -7.75
Colchicin - -8.06
Celecoxib -11.50 -
Docking 82 -Cox2
Docking 82 – tubulin
Docking 102 - COX2 Docking 102 -tubulin
(25)Designed compounds
Ligand efciency (LE)
COX2 Tubulin ΔLE
102 - 0.38 - 0.26 -0.12
82 - 0.38 - 0.30 -0.08
Combretatin A4 - 0.42 - 0.34 -0.08
Colchicin - - 0.28 -
Celecoxib - 0.44 - -
CONCLUSION
The thesis has achieved the following main results:
1 Successful design and synthesis of 41 multi target coxib - combretastatin hybrids In which, there are 20 coxib - combretastatin ester hybrid compounds, 20 acid coxib - combretastatin hybrid compounds, 01 coxib - combretastatin hybrid compound contains CF3 group
2 Evaluated biological activity of 22 substances on human cancer cell lines HT-29, Hep-G2, MCF-7 and inhibited NO production In which 12 substances 102, 96, 81, 82, 94, 93, 91, 92, 89, 90, 97, 85 show outstanding activity compared with celecoxib radicals, 05 substances 102, 96, 81, 82, 94 has IC50 activity <10 µM
3 Evaluation of NO production inhibition and breast cancer MCF-7 activity against acid-based coxib - combretastatin hybrids and comparison of ester-based coxib - combretastatin hybrids However, the results showed that only 02 substances 118 and 114 exhibited cytotoxicity on the lines of MCF-7 stronger than celecoxib and 02 substances 116 and 111 are equivalent to inhibiting NO production of celecoxib
(26)5.Collected typical of compouds 82 and 102, which are bio-multi-target molecules, have outstanding activity in anticancer activity compared with combretastatin and anti-inflammatory compared to celecoxib
6 On the molecular docking model, it is confirmed that two compouds 82 and 102 show double activity with two purposes of tubulin and COX2 effects according to the design objectives of the thesis
7 The substances obtained were shown to be non-toxic to normal cells in an in vitro model
Request: Further research into the anticancer and anti-inflammatory activity of these hybrid compounds has been obtained and clinical trials for the introduction of new anti-cancer drugs
NEW CONTRIBUTIONS OF THE DISSERTATION
1 Successful design and synthesis of 41 multi target coxib - combretastatin hybrids
2 Screening anti-inflammatory and anti-proliferative activity of the news hybrids
3 Evaluating of biological mechanism of the hybrid compounds in vitro Determinning interaction affinity of the hybrid on tubulin and COX2 by
(27)PUBLICATIONS
1 Thuy Hang Nguyen Thi, Yen Tran Thi, Le Anh Nguyen, Ngoc Binh Vo, Quo Anh Ngo “Design, Synthesis and Biological Activities of New Pyrazole Derivatives Possessing Both Coxib and Combretastatins Pharmacophores”
Chem Biodiversity, 2019
2 Quoc Anh Ngo, Thuy Hang Nguyen Thi, Minh Quan Pham, Domenico De fno hi hao Do “Antiproliferative and antiinfammatory coxib– combretastatin hybrids suppress cell cycle progression and induce apoptosis of MCF7 breast cancer cells” o e u ar Diversity 2020
3 Nguyen Thi Thuy Hang, Nguyen Le Anh, Tran hị Yen, Vo Ngọ Binh, Vu Đinh Hoang, Ngo Quoc Anh “Synthesis study of hybrid derivatives between combretastatin and celecoxib” Vietnam Journal of Chemistry, 2017, 55(4E23), 235-239
National Library of Vietnam