Translate this page into:
Assessment of two 1,2,3 triazole-related compound derivatives as having antiangiogenic property using cultured aorta ring and chorioallantonic membrane assay
*Corresponding author: Huda Ghassan Hameed, Department of Pharmacology, College of Pharmacy, Alshaab University, Baghdad, Iraq. huda.ghassan@alshaab.edu.iq
-
Received: ,
Accepted: ,
How to cite this article: Hameed HG, Sahib HB, Omran ZS. Assessment of two 1,2,3 triazole-related compound derivatives as having antiangiogenic property using cultured aorta ring and chorioallantonic membrane assay. Indian J Physiol Pharmacol. 2025;69:222-31. doi: 10.25259/IJPP_54_2024
Abstract
Objectives:
In this work, two 1,2,3 triazole related compounds (designated as H1 and H4) will be examined for their impact against new blood formation using an in vivo experimental model of forming a chorionic plexus in the chorioallantoic membrane (CAM) and an ex vivo sprouting microvessel model using rat aorta rings. Additionally, the mechanism behind their anti-angiogenic action will be explored. Inhibiting angiogenesis can be a valuable treatment avenue for cancer disorders because new blood vessel formation will facilitate tumor growth, invasion, and metastasis.
Materials and Methods:
Using the rat aortic ring (RAR) Assay, the rat aorta was divided into 1-mm sections as rings, were added to a prepared growth medium with specific additives to each well. Both synthesized 1,2,3 triazole related derivatives each alone was added at different dosages. The study proceeded with applying the CAM Assay where 10 mg/ml of each derivative was added and tested separately. A digital camera was used to take pictures of each CAM, and the blood vessel dimensions were measured digitally.
Results:
The RAR assay demonstrated that both new chemical entities inhibited angiogenesis in a concentration-effect dependent relationship. The CAM results demonstrated its significant blood vessel disruption action.
Conclusion:
This study showed that the derivatives of 1,2,3 triazoles significantly hinder the angiogenesis process, and could be classified as angiogenesis inhibitors.
Keywords
Angiogenesis
Chorioallantoic
Endothelial cells
1
2
3 triazole derivatives
INTRODUCTION
Angiogenesis is an essential physiologically controlled process that occurs under healthy conditions, for example, the menstrual cycle wound healing and embryogenesis. It is involved in many pathological conditions such as connective tissue disease, psoriasis, diabetic retinopathy, vascular ischemia and diseases of the blood vessels.[1]
Angiogenesis is a multi-step process, which includes firstly an angiogenic stimulation characterised by an increase in endothelial cell permeability, cell proliferation and capillary sprout length. Second, proteolyzaion of the basement membrane’s matricellular components is due to the activation of matrix metalloproteinases. Third, the development of tubes due to the endothelial cell multiplication and migration outside of the pre-existing capillary wall, and finally, the development of the basement membrane, adherent junctions and endothelial cells stabilise the capillary.[2]
It has been found that suppression or inhibition of angiogenesis using different therapeutic modalities is a very beneficial approach in the management of malignancies such as leukaemia and vascular aneurysms.[3,4] Previous research has shown that tumours can only grow and die up to 1–2 mm3 in diameter in the absence of angiogenesis due to lacking nutrients and oxygen, while in angiogenesis-rich cell culture, some tumour cells can grow and divide up to more than 2 mm3 in diameter.[5] Therefore, angiogenesis development is regulated by angiogenesis inducers and inhibitors.[6]
Herbs have long been used to cure cancer; in fact, traditional medicines for treating a wide range of illnesses have their roots primarily in the use of herbs. Researchers still view plants as crucial sources for the discovery of novel therapeutic factors, even though the actual molecules obtained from plants are not typically used as pharmaceuticals such as Phoenix dactylifera Seeds,[7] Cuminum cyminum seeds extract,[8] Ziziphus spina-christi leaves extracts,[9] Chalcones[10] and Prunus dulcis seed oil.[11] There are many medications that overcome the angiogenesis process at different cellular levels and they are potentially of benefit in cardiovascular and malignant diseases such as leukaemia and vascular aneurysms.[3,4] Heterocyclic compounds, including triazoles, are commonly investigated in the field of medical chemistry.[12,13]
The core of triazoles is three nitrogen, two carbons, and five-membered nitrogen atoms, which significantly participate in cellular activity.[14] The physiochemical properties of the 1,2,3-triazoles are aromatic compounds, having hydrogen binding properties and they are stable, that is resist acidic or basic hydrolysis and resist oxidising or reducing conditions even at high temperatures.[15] From a chemical point of view, triazoles are able to form a variety of non-covalent interactions that target many biological systems or organs through hydrophobic interactions, hydrogen bonds, van der Waals forces and dipole-dipole bonds.
Several studies found that 1,2,3-triazole derivatives have antibacterial,[16,17] antimalarial,[18] antiviral[19,20] and anticancer effects.[21,22]
A lot of pharmaceuticals that having 1,2,3-triazole core are licensed as anticonvulsants (rufinamide), antibiotics (cefatrizine), lactamase inhibitors (tazobactam) and anticancer (carboxyamidotriazole).[23]
There is evidence that medicines with anti-inflammatory properties can inhibit the angiogenesis process through several mechanisms.[24]
The rationale for this study is to investigate the antiangiogenic properties of N-benzyl phenyl isoserine derivatives [Figure 1] are related to the 1,2,3 triazoles and they have shown anti-inflammatory properties.[25,26] Therefore, this study aimed to look for the antiangiogenic properties of these derivatives using experimental models of aortic rings culture and chorioallantoic membrane assay.
- Chemical structure of N-benzyl phenyl isoserine derivatives.
MATERIALS AND METHODS
This research was conducted at the Department of Pharmacology, College of Medicine, Al-Nahrain University from 1 October 2022 to 1 April 2023. The study was approved by the Scientific and Ethical Committees at the College of Medicine, University of Al-Nahrain (No. 107, Date: 23 November 2022).
The derivatives will be referred to in the article as H1 & H4. Both compounds were generously provided by the Department of Pharmaceutical Chemistry, College of Pharmacy, University of Al-Kafeel, Najaf, Iraq. The chemical structures of the H1 are (3-benzamido-2-(4-((4-formylphenoxy)methyl)-1H-1,2,3-triazol-1-yl)-3-phenylpropanoic acid) [Figure 2] and of the H4 are (3-benzamido-2-(4-((4-nitrophenoxy)methyl)-1H-1,2,3-triazol-1-yl)-3-phenylpropanoic acid) [Figure 3]. The structure of the compounds was characterised by Fourier transform infrared analysis (FTIR) and 1H nuclear magnetic resonance analysis (NMR). These new chemical entities were dissolved in dimethyl sulfoxide and diluted with the M199 medium.
- Chemical structure of H1 compound.
- Chemical structure of H4 compound.
In this study, two experimental models for angiogenesis were used:
Rat aorta ring and angiogenesis assay
A modified technique of that mentioned by Brown et al.[27] was used for the preparation of aortic rings. Male Wistar rats of 5-week-old were anaesthetised using diethyl ether anaesthesia, animals were sacrificed by cervical dislocation.
The fibro-adipose tissues were removed from the thoracic aorta, washed with serum-free medium and cross-sections of 1 mm thickness were obtained.
The tissue specimens were cleaned of peri adventitial fibro adipose material and residual blood clots, this was then cut into 1 mm thick aortic ring segments which were exposed to the medium M199 and it was immersed in a solution composed of fibrinogen and aprotinin at concentrations of 3 mg/mL and 5 mg/mL, respectively. One aortic ring was then placed in each 48-well plate filled with 300 μL of (3 mg/mL) fibrinogen and (5 mg/mL) of Aprotinin in medium (M199) supplemented with 20% heat inactivated fetal calf serum (HIFCS), 0.1% έ-aminocaproic acid, 1% L-Glutamine, 1% amphotericin and 0.6% gentamicin were added to each well and ring tissues were placed in the centre of the well, followed by 15 μL of thrombin (50 National Institutes of Health [NIH] U/mL) in 0.15 M NaCl. Immediately after embedding the vessel fragment in the fibrin gels, 0.5 mL of medium M199, the compound H1 and H4 each alone were applied to the upper layer at different concentrations ranging between 12.5 and 200 μg/mL. Then, the samples were incubated in an incubator at 37°C and supplemented with 5% carbon dioxide (CO2). After 72 h of incubation, the top layer was washed with M199 media and evidence of angiogenesis was detected as the formation of new vessels using a camera and ImageJ’s apparatus (https://imagej.net/Fiji/ Downloads software program). The inhibitory effect of the H1 compound was determined using the method introduced by Nicosia et al.[28]
The following formula was used to determine the percentage of blood vessel inhibition:
Blood vessels inhibition (%) = 1 − (A0/A) × 100
Where: A0: The distance of the test substance’s blood vessel growth in millimetres; A: The distance of the negative control’s blood vessel growth in millimetres. The distance was determined using an ordinary microscope at 40 magnification power and the negative control sample represented the aortic ring treated with the solvent used to dissolve the tested compounds (dimethyl sulfoxide, 1% v/v).
Quantification and imaging of rat aortic ring (RAR) assay analysis
Radial spacing between the sprouts: We first collected live specimens phase contrast pictures, followed by utilizing the rolling ball radius of 700 pixels surrounding the aortic piece to abolish the backdrop from each image using ImageJ’s subtract background function (https://imagej.net/Fiji/ Downloads). This process improved contrast and reduced noise. Subsequently, the sprouts were highlighted exclusively, while the aortic ring and any empty spaces were erased or ignored using the ‘alter threshold’ option.[29]
Chorioallantois membrane (CAM) in vivo assay
Fertilised chicken eggs from the veterinary medicine college at Baghdad University poultry’s grounds were collected and cleaned using filth with 70% ethanol, then put in a CO2 incubator for 72 h at 37°C with a humidity of about 60%. The eggs were positioned horizontally and rotated repeatedly. The eggs were incubated for another 24 h, followed by sucking out 2 mL of albumin and closing the tiny hole that had been perforated down by the side ‘to provide better visualisation of the formed CAM, where the CAM will detach from the egg shell.’ Following the creation of a tiny square window of about (3–4 cm in length diameter) in the eggshell, test samples that had been pre-soaked in rounded discs of filter paper were placed on the CAM and the window was sealed with antiseptic surgical adhesive tape. After that, the eggs were incubated for another 72 h. The zone of blood vessel disruption or inhibition had been calculated and photographed.
The test compounds (H1 and H4) were prepared as 10 mg/mL, 50 μL (the final dose was 0.5 mg/disc of each compound). These samples were then placed alone on a 6 mm Whitman filter paper disc to be coated with them before being allowed to dry at 45–50°C. The loaded and dried discs were placed on the CAM. The digital camera was used to take pictures of each CAM and record its dimensions. The entire procedure was carried out in an aseptic environment.[30]
Analysis and imaging of CAM
Tools for studying angiogenesis using images have been successfully developed. According to Mutka and Bart,[31] image-based phenotyping can overcome some of the drawbacks of the more conventional techniques mentioned above. While the ability to handle image, data has improved thanks to machine learning, good trait classification or quantification frequently requires huge datasets, which can be difficult to obtain. More affordable, few-shot image analysis technologies are, therefore, required to efficiently segment and quantify illness symptoms. In this study, we used the Fiji version of ImageJ to investigate the vessel network. Images were opened and the background was corrected. Both the manual and ImageJ’s Skeletonize (2D/3D) plugins were utilised in the Fiji (Fiji is Just ImageJ) edition of the software.[32]
Quantification and grading for the responses were + (3–6 mm), ++ (6–9 mm) and +++ (>10 mm). The findings were displayed as mean ± standard deviations of means. An image analyser (BIOCOM Visiolab TM 2000) was used to quantify the inhibitory zone.[33]
Statistical analysis
The data were presented as numbers, percentages and whenever possible as mean and standard deviation. The mean value of six experiments at each concentration of H1 or H4 compounds was used in plotting the best-fit line of the log concentration curve. The X-axis represents the log concentration of each compound, while the Y-axis represents the percentage of inhibition. The log concentration-inhibitory effect (%) was created to estimate the inhibitory effect of each compound at a specific concentration using a Pearson’s correlation test with a best-fit line regression analysis. The IC50 (Inhibitory Concentration 50%) of each compound was calculated using a regression equation by adjusting 50% at the y value. The relative potency of H1-to-H4 was calculated by dividing the percentages of growth inhibition (%) of H1 by H4 at specific fixed concentrations for both H1 and H4. An independent two-sample, two-tailed t-test was used to test the significant differences in the inhibitory effects of H1 and H4. P < 0.05 is the lowest limit of significance. The Excel 2010 program was used for computing the data (Microsoft Cooperation, Redmond, Washington, U.S.).
RESULTS
Rat aorta ring experimental model
Tables 1 and 2 showed that both compounds inhibit angiogenesis by more than 50% at any concentration ranging between 12.5 and 200 μg/mL. The efficacy of the H1 compound is significantly higher than that of H4 in inhibiting angiogenesis. The maximum mean inhibitory effect of both compounds was 69% at a concentration of 200 μg/mL. The results revealed that all concentrations of H1 and H4 compounds each alone in the ongoing study retained the antiangiogenic activity. A higher percentage of angiogenesis inhibition was observed at a concentration of 200 μg/mL. These findings indicated that both compounds inhibited the growth of the blood vessels in a concentration-effect dependent. The maximum inhibition was observed at a concentration of 200 μg/mL, while the serial dilutions of these compounds at concentrations of <200 μg/mL produced variable inhibitory effects.
| Concentration μg/mL | Percentage of inhibition±SD |
|---|---|
| 200 | 69±1.8 |
| 100 | 69±3.4 |
| 50 | 62±1.1 |
| 25 | 58±0.9 |
| 12.5 | 55±0.3 |
SD: Standard deviation
| Concentration μg/mL | Percentage of inhibition±SD |
|---|---|
| 200 | 69±0.4 |
| 100 | 61±3.36 |
| 50 | 50±6.2 |
| 25 | 50±0.9 |
| 12.5 | 51±3.65 |
SD: Standard deviation
From the linear regression equation, the IC50 of H1 and H4 compounds were calculated given the values of 0.91 and 1.16 μg/mL, respectively [Figure 4]. Therefore, H1 is more potent than H4 by 1.3-fold.
- Comparison between H1 and H4 compounds on the inhibition of new vascularisation using the rat aortic ring assay. Blue line represents the inhibition of blood vessel growth by H1, while the orange line represent inhibition by H4.
Microscopic finding
Figures 5a and 6a showed that H1 and H4 compounds inhibited the microvasculature outgrowth at IC50 of 0.91 and 1.16 1.16 μg/mL, respectively, compared with untreated aortic rings. This observation seems to be significantly (P < 0.05) concentrations-dependent as illustrated in Figures 5c and 5d, 6c and d, respectively. 50% of inhibition of vascular formation was observed at a concentration of 100 μg/mL and a higher concentration of 200 μg/mL produced complete inhibition.
- (a) Untreated control showing normal angiogenesis with complete vascularization, (b-f) Treated are sample with (6.25, 12.5, 25, 50, 100, and 200 µg/mL H1 showing partial inhibition of vascular sprouting demonstrating concentration related significant disruption of microvascular formation.
- Rat aortic ring assay images of control and 24 h treated with different concentrations of H4. (a) Untreated control showing normal angiogenesis with complete vascularization. (b-f) Treated sample with (6.25, 12.5, 25, 50, 100, and 200 µg/mL H1 showing partial inhibition of vascular sprouting demonstrating concentration related significant disruption of microvascular formation. Research laboratory and the obtaining results are similar to more sophisticated models.
Chorioallantoic membrane (CAM) experimental model
Both compounds inhibit the angiogenesis process as there is a reduction in the length of the blood vessels (mm), which is detected as the efficacy of the H1 compound is not significantly (P = 0.312) lower than that of the H4 compound in inhibiting new blood vessel formation. The regression in blood vessel growth was assessed using the previously indicated scoring system. Both chemical entities achieved a considerable score (+++), that is the blood vessel formation is inhibited by >10 mm. A reduction of the blood vessel length ranged between 9–13 and 9–15 mm for H1 and H4 compounds, respectively, as shown in Tables 3 and 4. The results of the CAM study showed inhibition of angiogenesis and a distortion of the vascular architecture. Figures 7 and 8 showed a significant decrease in the number of blood vessels due to the effects of H1 and H4, respectively. Both new chemical entities significantly inhibited angiogenesis, indicating that they have antiangiogenic properties in vivo. The potency of H4 is a little bit higher than H1. This observation supports the results obtained with the ex vivo and in vitro studies.
| Egg no. | Length of inhibited blood vessels (mm) | Inhibition scoring |
|---|---|---|
| 1 | 9 | ++ |
| 2 | 11 | +++ |
| 3 | 12 | +++ |
| 4 | 11 | +++ |
| 5 | 13 | +++ |
| 6 | 9 | ++ |
| Mean±SD | 10.6±1.86 | +++ |
CAM: Chorioallantoic membrane, SD: Standard deviation. The grading system: + (3 - 6 mm), ++ (6 - 9 mm), and +++ (> 10 mm), reflecting the size of the inhibitory zone
| Egg no. | Length of inhibited blood vessels (mm) | Inhibition scoring |
|---|---|---|
| 1 | 15 | +++ |
| 2 | 11 | +++ |
| 3 | 12 | +++ |
| 4 | 11 | +++ |
| 5 | 13 | +++ |
| 6 | 9 | ++ |
| Mean±SD | 11.83±2.04 | +++ |
CAM: Chorioallantoic membrane, SD: Standard deviation. The grading system: + (3 - 6 mm), ++ (6 - 9 mm), and +++ (> 10 mm), reflecting the size of the inhibitory zone
- Chick chorioallantoic membrane assay images of control and 24-h-treated Eggs with H1 compound; (a and c) control chorioallantois membrane (CAM) showing normal blood vessel sprouting and branching, indicating physiological angiogenesis. (b and d) CAM treated with H1 compound for 24 h, demonstrating disruption of blood vessel formation and a reduction in vessel branching, indicative of the compound’s antiangiogenic activity.
- Chick chorioallantoic membrane assay images of control and 24-h-treated eggs with H4 compound; (a and c) control chorioallantois membrane (CAM) showing normal blood vessel sprouting and branching, indicating physiological angiogenesis. (b and d) CAM treated with H4 compound for 24 h, demonstrating disruption of blood vessel formation and a reduction in vessel branching, indicative of the compound’s antiangiogenic activity.
DISCUSSION
1,2,3-Triazoles are one of the most important classes of nitrogen-containing heterocycles. Cefatrizine derivatives, carboxyamido-triazoles, azido-β-lactam, amprenavir derivatives, 1,2,3-triazole-dithiocarbamate-urea hybrid and N-((1-benzyl-1H-1,2,3-triazole-4-yl)methyl) arylamide derivatives are 1,2,3-triazole-containing compounds that demonstrated anticancer action.[34] Studying the molecular mechanism of the compounds was not in our objectives of the study, but the chemical structure of the compound, including the position and orientation of the aldehyde group may contribute to its enhanced inhibitory activity. Certain 1,2,3-triazole derivatives have been shown in previous studies to mitigate mitochondrial dysfunction, ameliorate hypoxia-induced oxygenation damage and reduce vascular dysfunction.[34] H1 compound could be similarly act like these compound that have been studied. Further investigation to confirm that are needed.
The current work shows that these new chemical entities inhibit the angiogenesis process in two experimental models. The H1 compound is significantly more efficient than the H4 compound in inhibiting the angiogenesis process on the RAR, but it has the same inhibitory effect as the H4 on the CAM. Therefore, both compounds are in favour of inhibiting angiogenesis in tumour growth or metastasis, while the H1 compound may be a preferable new chemical entity for inhibiting angiogenesis in cardiovascular disorders. In this study, two models of angiogenesis were used to ascertain the anti-angiogenic properties of two new chemical entities. Most recent studies used mouse aortic rings for studying specific molecular changes, but the RAR model is still used for screening anti-angiogenic molecules.[35] These models provide valuable insights into the biological effects of compounds and their potential therapeutic applications. One commonly used in vivo model for angiogenesis is the RAR assay, which involves culturing aortic rings in a gel matrix.[36] This assay allows the observation of sprouting blood vessels from the aortic rings.
Another frequently employed in vivo model is the CAM assay, which utilises the embryonic membrane of a developing chicken egg. The CAM assay allows direct visualisation of blood vessels and their response to angiogenic stimuli.[37] The RAR assay and CAM assay are commonly used experimental approaches to evaluate the anti-angiogenic activity of compounds or substances.[36,38] We believe that using such model is feasible, available in any research laboratory and the obtaining results are similar to more sophisticated models.
The discrepancy in efficacy between H1 and H4 in both models could be related to: Their effects on different receptors or metabolic pathways in each experimental model. Hence, it can be suggested that H1 compound has pleiotropic effects.
Previous studies have proven that the presence of heteroatoms directly affects their toxicity against cancer cells.[39] 1,2,3-Triazole hybrids have attracted interest from all around the world. In the field of organic and medicinal chemistry, this lead chemical has the potential to be a highly effective therapeutic candidate.[40] In 2021, Wang et al.[41] study 1,2,3-triazole scaffolds derivatives, and the results shows inhibitory activity on vascular endothelial growth factor receptor-2 (VEGFR-2) with one derivative having better anti-angiogenesis ability than sunitinib.
The H1 compound has been found to be more potent than the H4 compound, indicating that H1 could act as an anti-tumour because most studies used the RAR model to assess angiogenesis by demonstrating inhibition of the formation of microvessels that are present in malignancies or their metastasis. Therefore, it is possible that the H1 compound induces antiangiogenic properties through different mechanisms, including attenuation of angiogenesis sprouting;[42] reducing new vessel formation[43] and reducing the expression of vascular and basic fibroblast growth factors.[44] Hence, the anti-angiogenesis of these compounds could be attributed to the inhibition of proteolytic damage to the basement membrane of the capillary vessels, proliferation, migration, induction of endothelial cell tubulogenesis, vessel formation and pruning and pericellular stabilisation.[45] The above effects are the results of the H1 and H4 acting on certain receptors or metabolic pathways, which our opinion suggested that they act on the Integrin receptors: Integrins are cell surface receptors that mediate cell adhesion and signalling. They are involved in angiogenesis by regulating endothelial cell migration and adhesion to the extracellular matrix,[46] or might be the action of specific metabolic pathways. However, it is challenging to determine which mechanism may play a predominant role in the observed anti-angiogenic effects; it is important to conduct further investigations to elucidate the drug’s mechanism of action.
Therefore, these compounds act directly on the endothelial cells of the RAR, as previous studies demonstrated that triazoles inhibit angiogenesis in the above-mentioned different steps of angiogenesis.[47] The variability of these compounds in producing maximum inhibition of angiogenesis is related to the critical factors that affect the RAR assay, which included the animal species (rats responded to angiogenesis less than mice) and sex (females responded less than males), the culture media and the addition of growth factors.[48]
The second model used in this study was the CAM. It is a non-specific model for angiogenesis as it was used in different fields, for example, photodynamic therapy.[49] Therefore, it is expected to find that there is a non-significant difference between H1 and H2 chemical entities in angiogenesis, and it can be postulated that these compounds were acting on angiogenesis that is related to cardiac biology.[50] Recent studies improved the critical use of this model to study tumour genesis and cancer chemotherapy by inoculating patients’ tumour cells into CAM to produce tumours in chicken eggs, which is not done in this study, and it is an important limitation of this work.[51] Because angiogenesis is a complex biological process, different assays provide complementary information that helps researchers gain a more comprehensive understanding. Assays such as the CAM assay, the RAR assay and the endothelial cell tube formation assay are designed to mimic specific physiological conditions or tissue types.[37] Some variations can occur due to the inherent differences between assays, including variations in experimental conditions, model systems and the specific endpoints being measured, but aligning results sure gives strength to the conclusion.
Therefore, both H1 and H4 reduced the length of new blood vessels through different mechanisms related to environmental conditions, for example, oxygen deprivation, glucose supplement, and reactive or nitrative oxygen species.[52-54] In our work, all the environmental conditions were adjusted; therefore, these new chemical entities were acting directly on the formation of new vessels. Literature surveys do not disclose the effects of triazoles on the angiogenesis of the CAM model, but recent studies demonstrated that 1,2,3-triazoles inhibit VEGFR-2, leading to suppression of cellular growth in the cultured cancer cell.[55]
Limitation of the study
Limitations of our study include the absence of inoculation of patients’ tumour cells into CAM to produce tumours, and the lack of techniques such as Western blotting and gene expression analysis to investigate the underlying molecular mechanism. Without a clear understanding of the underlying mechanism, it is challenging to determine whether the observed effects in the RAR and CAM assays could be translated to other angiogenesis-related processes or disease models. The above-mentioned limitations will not negatively impact our results regarding the angiogenesis process, but it is absolutely required in studying the effects of these compounds on the tumour cell.
CONCLUSION
1,2,3-triazole derivatives are new chemical entities that inhibit the angiogenesis process in both experimental animal models and will be promising molecules for tumorigenesis, particularly H1 compound, and in pathological conditions associated with angiogenesis. According to the animal model, these molecules act directly on the angiogenesis process.
Acknowledgement:
We would like to express our sincere gratitude to all individuals who supported us throughout the completion of this study.
Ethical approval:
The research/study was approved by the Institutional Review Board at College of Medicine/Al-Nahrain University, approval number 20200969, dated 23rd November 2022.
Declaration of patient consent:
Patient’s consent was not required as there are no patients in this study.
Conflicts of interest:
There are no conflicts of interest.
Use of artificial intelligence (AI)-assisted technology for manuscript preparation:
The authors confirm that there was no use of artificial intelligence (AI)-assisted technology for assisting in the writing or editing of the manuscript and no images were manipulated using AI.
Financial support and sponsorship: Nil.
References
- Understanding angiogenesis and its clinical applications. Ann Clin Lab Sci. 2000;30:227-38.
- [Google Scholar]
- Angiogenesis and its therapeutic opportunities. Mediators Inflamm. 2013;2013:127170.
- [CrossRef] [PubMed] [Google Scholar]
- Angiogenesis in cancer and other diseases. Nature. 2000;407:249-57.
- [CrossRef] [PubMed] [Google Scholar]
- Plaque neovascularization and antiangiogenic therapy for atherosclerosis. J Am Coll Cardiol. 2007;49:2073-80.
- [CrossRef] [PubMed] [Google Scholar]
- The good, the bad, and the ugly: Neutrophils, angiogenesis, and cancer. Cancers (Basel). 2022;14:536.
- [CrossRef] [PubMed] [Google Scholar]
- Antiangiogenic activity of sweet almond (Prunus dulcis) oil alone and in combination with aspirin in both in vivo and in vitro assays. Asian Pac J Cancer Prev. 2022;23:1405-13.
- [CrossRef] [PubMed] [Google Scholar]
- The anti-angiogenic activity of Phoenix dactylifera seeds methanol extract in vivo study. Iran J Pharm Sci. 2018;14:83-92.
- [Google Scholar]
- Anti-angiogenic activity of Cuminum cyminum seeds extract: In vivo study. Karbala J Pharm Sci. 2017;8:1-9.
- [Google Scholar]
- Anti-angiogenic activity of Ziziphus spina-christi leaves extracts. Int J Pharm Sci Rev Res. 2015;35:32.
- [Google Scholar]
- The anti-angiogenic activity of p-hydroxy chalcone. Int J Pharm Sci Rev Res. 2016;37(1):117-121.
- [Google Scholar]
- Anti-proliferative activity of Prunus dulcis seed oil alone and in combination with aspirin on human umbilical vein endothelial (HUVECs) and Kaposi sarcoma (KS) cells. J Popul Ther Clin Pharmacol. 2023;30:220-31.
- [CrossRef] [Google Scholar]
- Pyrrole: An insight into recent pharmacological advances with structure activity relationship. Eur J Med Chem. 2018;157:527-61.
- [CrossRef] [PubMed] [Google Scholar]
- Synthesis of medicinally relevant scaffolds-triazoles and pyrazoles in green solvent ionic liquids. Curr Organ Chem. 2021;25:1523-38.
- [CrossRef] [Google Scholar]
- Synthesis of pharmacological relevant 1,2,3-triazole and its analogues-a review. Curr Org Synth. 2021;18:101-24.
- [CrossRef] [PubMed] [Google Scholar]
- Cu-catalyzed azide-alkyne cycloaddition. Chem Rev. 2008;108:2952-3015.
- [CrossRef] [PubMed] [Google Scholar]
- Comprehensive review on the anti-bacterial activity of 1,2,3-triazole hybrids. Eur J Med Chem. 2019;168:357-72.
- [CrossRef] [PubMed] [Google Scholar]
- Medicinal attributes of 1,2,3-triazoles: Current developments. Bioorg Chem. 2017;71:30-54.
- [CrossRef] [PubMed] [Google Scholar]
- A review on diverse heterocyclic compounds as the privileged scaffolds in antimalarial drug discovery. Eur J Med Chem. 2018;158:917-36.
- [CrossRef] [PubMed] [Google Scholar]
- Targeting the entrance channel of NNIBP: Discovery of diarylnicotinamide 1,4-disubstituted 1,2,3-triazoles as novel HIV-1 NNRTIs with high potency against wild-type and E138K mutant virus. Eur J Med Chem. 2018;151:339-50.
- [CrossRef] [PubMed] [Google Scholar]
- Dihydropyrimidinone/1,2,3-triazole hybrid molecules: Synthesis and anti-varicella-zoster virus (VZV) evaluation. Eur J Med Chem. 2018;155:772-81.
- [CrossRef] [PubMed] [Google Scholar]
- Recent advancements in 1,4-disubstituted 1H-1,2,3-triazoles as potential anticancer agents. Anticancer Agents Med Chem. 2018;18:21-37.
- [CrossRef] [PubMed] [Google Scholar]
- Structure-activity relationship (SAR) study and design strategies of nitrogen-containing heterocyclic moieties for their anticancer activities. Eur J Med Chem. 2017;125:143-89.
- [CrossRef] [PubMed] [Google Scholar]
- Click chemistry: 1,2,3-triazoles as pharmacophores. Chem Asian J. 2011;6:2696-718.
- [CrossRef] [PubMed] [Google Scholar]
- Inhibition of angiogenesis by non-steroidal anti-inflammatory drugs: From the bench to the bedside and back. Curr Drug Targets Inflamm Allergy. 2005;4:31-8.
- [CrossRef] [PubMed] [Google Scholar]
- Design, synthesis anticytokine storm and molecular docking studies of N-benzyl phenyl isoserine derivatives containing 1, 2, 3-triazoles. Azerbaijan Med J. 2022;62(05)
- [Google Scholar]
- In silico design, and in vivo evaluation of synthesized phenyl isoserine derivative and used as anticytokine storm. Egypt J Chem. 2023;66:151-9.
- [Google Scholar]
- Endogenous regulation of angiogenesis in the rat aorta model. Role of vascular endothelial growth factor. Am J Pathol. 1997;151:1379-86.
- [Google Scholar]
- A simplified aortic ring assay: A useful ex vivo method to assess biochemical and functional parameters of angiogenesis. Matrix Biol Plus. 2020;6-7:100025.
- [CrossRef] [PubMed] [Google Scholar]
- Anti-angiogenic activity of Matricaria chamomilla flowers methanol extract-in vivo study. Int J Pharm Sci Rev Res. 2016;41:367-71.
- [Google Scholar]
- Image-based phenotyping of plant disease symptoms. Front Plant Sci. 2014;5:734.
- [CrossRef] [PubMed] [Google Scholar]
- 3D reconstruction of histological sections: Application to mammary gland tissue. Microsc Res Tech. 2010;73:1019-29.
- [CrossRef] [PubMed] [Google Scholar]
- Chick chorioallantoic membrane (CAM) assay as an in vivo model to study the effect of newly identified molecules on ovarian cancer invasion and metastasis. Int J Mol Sci. 2012;13:9959-70.
- [CrossRef] [PubMed] [Google Scholar]
- 1, 5-disubstituted-1, 2, 3-triazoles counteract mitochondrial dysfunction acting on F1FO-ATPase in models of cardiovascular diseases. Pharmacol Res. 2023;187:106561.
- [CrossRef] [PubMed] [Google Scholar]
- The aortic ring assay and its use for the study of tumor angiogenesis. Methods Mol Biol. 2016;1464:63-72.
- [CrossRef] [PubMed] [Google Scholar]
- The aortic ring model of angiogenesis: A quarter century of search and discovery. J Cell Mol Med. 2009;13:4113-36.
- [CrossRef] [PubMed] [Google Scholar]
- Microvascular experimentation in the chick chorioallantoic membrane as a model for screening angiogenic agents including from gene-modified cells. Int J Mol Sci. 2021;23:452.
- [CrossRef] [PubMed] [Google Scholar]
- Quantitation of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane. Microvasc Res. 1994;47:31-40.
- [CrossRef] [PubMed] [Google Scholar]
- 1,2,3-Triazole-containing hybrids as leads in medicinal chemistry: A recent overview. Bioorg Med Chem. 2019;27:3511-31.
- [CrossRef] [PubMed] [Google Scholar]
- A recent overview of 1, 2, 3-triazole-containing hybrids as novel antifungal agents: Focusing on synthesis, mechanism of action, and structure-activity relationship (SAR) J Chem. 2022;2022:1-50.
- [CrossRef] [Google Scholar]
- Design, synthesis, and in vitro and in vivo anti-angiogenesis study of a novel vascular endothelial growth factor receptor-2 (VEGFR-2) inhibitor based on 1,2,3-triazole scaffold. Eur J Med Chem. 2021;211:113083.
- [CrossRef] [PubMed] [Google Scholar]
- Diosmetin inhibits tumor development and block tumor angiogenesis in skin cancer. Biomed Pharmacother. 2019;117:109091.
- [CrossRef] [PubMed] [Google Scholar]
- Myricetin inhibits breast tumor growth and angiogenesis by regulating VEGF/VEGFR2 and p38MAPK signaling pathways. Anat Rec (Hoboken). 2019;302:2186-92.
- [CrossRef] [PubMed] [Google Scholar]
- Catalpol suppressed proliferation, growth and invasion of CT26 colon cancer by inhibiting inflammation and tumor angiogenesis. Biomed Pharmacother. 2017;95:68-76.
- [CrossRef] [PubMed] [Google Scholar]
- The role of integrins in inflammation and angiogenesis. Pediatr Res. 2021;89:1619-26.
- [CrossRef] [PubMed] [Google Scholar]
- Carboxyamido-triazole inhibits angiogenesis by blocking the calcium-mediated nitric-oxide synthase-vascular endothelial growth factor pathway. J Pharmacol Exp Ther. 2000;292:31-7.
- [CrossRef] [PubMed] [Google Scholar]
- Critical factors in measuring angiogenesis using the aortic ring model. J Genet Syndr Gene Ther. 2013;4:1000147.
- [Google Scholar]
- The chicken embryo chorioallantoic membrane as an in vivo model for photodynamic therapy. Methods Mol Biol. 2022;2451:107-25.
- [CrossRef] [PubMed] [Google Scholar]
- The chick embryo as an expanding experimental model for cancer and cardiovascular research. Dev Dyn. 2014;243:216-28.
- [CrossRef] [PubMed] [Google Scholar]
- Patient derived chicken egg tumor model (PDcE Model): Current status and critical issues. Cells. 2019;8:440.
- [CrossRef] [PubMed] [Google Scholar]
- Methylglyoxal scavengers attenuate angiogenesis dysfunction induced by methylglyoxal and oxygen-glucose deprivation. Oxid Med Cell Longev. 2022;2022:8854457.
- [CrossRef] [PubMed] [Google Scholar]
- Reduced O2 concentration during CAM development--its effect on angiogenesis and gene expression in the broiler embryo CAM. Gene Expr Patterns. 2012;12:236-44.
- [CrossRef] [PubMed] [Google Scholar]
- High-affinity Cu(I)-chelator with potential anti-tumorigenic action-a proof-of-principle experimental study of human h460 tumors in the CAM assay. Cancers (Basel). 2022;14:5122.
- [CrossRef] [PubMed] [Google Scholar]
- The effect of novel synthetic semicarbazone-and thiosemicarbazone-linked 1,2,3-triazoles on the apoptotic markers, VEGFR-2, and cell cycle of myeloid leukemia. Bioorg Chem. 2022;127:105968.
- [CrossRef] [PubMed] [Google Scholar]