New biocompatible products shagaol and curcuminoid-like type used as adjuvantes in cancer radiotherapy

CANRADIOPROTECT

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Contracting authority: Executive Unit for the Financing of Higher Education, Research, Development and Innovation – UEFISCDI

Project number: PN-III-P2-2.1-PED-2019-1471

Partners: National Institute for Research & Development in Chemistry and Petrochemistry –ICECHIM

University of Bucharest, The Faculty of Biology

Contract number: 363PED/2020

Implementation period: 2020-2022

Budget: 600.000 lei



About

The proposal has as a general objective the obtaining of keto-enolic derivatives of natural origin and of synthesis. This project refers to an interdisciplinary field of research and aims to elaborate methods of synthesis and biological testing of new compounds, symmetrical β-di-ketones (curcumin derivatives) and asymmetric mono or di-ketones (shagaol derivatives). After morphostructural characterization, biocompatibility testing and establishing the structure-property relationship, biologically active products will be applied in the radiotherapy treatment of cancer. In this way it is expected that the new structures synthesized, analogues of shagaol and curcumin, have higher antitumor activity than that of natural compounds and be used as adjuvants in cancer therapy.

Obiectives

O1. Selection of natural compounds and synthesis of biologically active chromogens (curcumin and shagaol) with free radical capture properties and

choice of demonstrative experimental model.

O2. The study of the radioprotective and radio sensitizing processes of the compounds selected after the biocompatibility tests.

O3. The correlation of the intensity of the effect of radioprotection and radiosensitization with the structural characteristics of the selected compounds.

Estimated results:

New products:

• 2 compounds derived from curcumin with radioprotective / radiosensitizing properties;

• 2 analogous shogaol compounds with radioprotective / radiosensitizing properties.

Obtaining technologies and methods of application:

Technology for obtaining keto-ethylene products with radioprotective / radiosensitizing properties.

Method and conditions of treatment for radiotherapy and radiation protection of whole erythrocyte.


The main results regarding the synthesis, characterization and radioprotective or radiosensitizing properties manifested by new keto-enol derivatives will be disseminated by nationally and internationally  publication in specialized scientific journals: 5 articles ISI, 6 scientific communications at international conference and 2 national patent applications. 

Project development

Stage 1 – 2020

Selection of natural compounds with antioxidant properties and elaboration of concept design of the experimental model.

Following the analysis of data published in the literature on active compounds extracted from turmeric, the team from INCD-ICECHIM (CO) starts the

first experimental activities aimed at the extraction by classical methods of maceration and decoction of curcumin derivatives from turmeric root.

The aqueous extracts obtained were analyzed by thin layer chromatography and UV-Vis spectroscopy, and the solid residue from the extraction process was analyzed by FTIR spectroscopy, reflectance spectroscopy and thermogravimetric analysis. These studies were performed in order to identify, isolate and characterize the main compounds found in natural extracts of Curcuma Longa (turmeric).

In parallel, the research team of the partner, University of Bucharest (UB), conducted testing studies on two different cell types, the HeLa tumor cervix cell line, compared to the MRC-5 non-tumor cell line, to be able to highlight a specificity of action on malignant cells.

An Isolera Prime flash chromatograph system was purchased at this stage of the project. This makes it possible to separate the compounds from the mixtures or to characterize the unitary compounds in terms of purity. The device is useful in the project for conducting studies to evaluate plant mixtures obtained under different extraction conditions and for the characterization of newly synthesized compounds in the process of their separation and purification.

Isolera Prime is a high performance flash chromatography system consisting of the following integrated components:

• A solvent-resistant, color LCD touch screen with a resolution of 800 x 600 pixels. It serves both as a display and as a system input device via the touch controls on the screen.

• A fraction collector.

• A pump module, which directs the flow of liquid through the system.

• An internal detector, which provides the system with information on the light absorbance of solvents and samples passing through the flow cell of the detector.

Stage 2 – 2021

Alternatives for obtaining natural or synthetic compounds, choosing the functional model and testing them in the processes involved in radiotherapy

In the second stage, studies on the methods of extraction and characterization of the main compounds from turmeric and ginger were continued by CO, using the soxhlet method, microwave field extraction and assisted extraction by ultrasonic processes, in water or alcohol. The natural extracts obtained were characterized for the identification of the main constituent compounds, by thin layer chromatography, and then the compounds were separated

by the ˮflash chromatographyˮ process. The separated fractions were analyzed by UV-Vis spectroscopy, and together with the residual mass resulting from the extraction processes were characterized by FTIR spectra and diffuse reflection spectra.

Similar to the structures of the natural compounds curcumin and shagaol, extracted from turmeric and ginger, respectively, a series of compounds were synthesized using the solvent method or the microwave field. Thus, bis-aryl-1,6-heptadiene-3,5-dionic chromogens obtained in the microwave field. While curcuminic derivatives, 3,5-bis (styryl) pyrazolonic compounds were synthesized by the classical method. In parallel, 6-aryl-5-hexene-2,4-dione derivatives were synthesized by solvent-free in the microwave field. The compounds resulted from the condensation reaction between substituted benzaldehydes with acetylacetone, hexanal. At the same time, 6-aryl-5-hexen-2-one structures were obtained by the conventional solvent method, following successive condensation reactions between substituted benzaldehydes with acetone and then with hexanal. The synthesized compounds were purified by specific methods and characterized by thin layer chromatography, FTIR spectroscopy, UV-Vis spectroscopy, NMR and thermogravimetric analysis.

The UB partner, following the concept design established in the first stage, tested the curcumin and shagaol derivatives, synthesized by CO, using

the two in vitro cellular models, for the biocompatibility and toxicity tests, respectively. In parallel, in order to establish the treatment technology, the

efficiency of blood cells as cell models for the evaluation of the properties exerted by curcumin derivatives was first tested. The method chosen to

determine the activity of the enzyme glutathione peroxidase GPx is based on monitoring the formation of GSSG (glutathione disulfide), which is continuously reduced by adding an excess of glutathione reductase, generating a constant level of GSH (Glutathione). After choosing the test method, from the batch of compounds analyzed in terms of biocompatibility and cytotoxicity, two compounds were selected that showed very low cytotoxicity. These beta-diketone derivatives with the styryl-pyrazolonic structure and the curcumin standard were subjected to tests performed by exposure to gamma radiation (10 or 50 Gy), regarding the evaluation of the radiation protection properties on erythrocytes. Samples of erythrocyte membranes resulting from erythrocyte lysis and removal of intracellular contents were analyzed by THz spectroscopy. The test results showed that in terms of radiation protection, 3,5-bis (styryl) pyrazolone appears to be the most effective.

Results

Papers

1.     Raduly, F.M.; Raditoiu, V.; Raditoiu, A.; Frone, A.N.; Nicolae, C. A.; Purcar, V.; Ispas, G.; Constantin, M.; Raut, I. “Modeling the properties of curcumin derivatives in relation to the architecture of the siloxane host matrices”, Materials, 2022, 15, 267, doi.org/ 10.3390/ma15010267.

2.    Raduly, F.M.; Raditoiu, V.; Raditoiu, A.; Purcar, V. “Curcumin: Modern Applications for a Versatile Additive”, Coatings, 2021, 11, 519, doi.org/10.3390/coatings11050519.

3.   Avram, S.; Stan, M.; Udrea A.M.; Buiu, C.; Boboc, A.A.; Mernea, M.  “3D-ALMOND-QSAR Models to Predict the Antidepressant Effect of Some Natural Compounds”. Pharmaceutics, 2021, 13.9, 1449, doi.org/10.3390/pharmaceutics13091449.

4.    Niculae, R.; Mernea, M.; Ghica, L.; Mihăilescu, D. F. ”A Computational Study on Ca2+ Modulation of ASIC 1 Pharmacologic Properties”. Studia Universitatis Babes-Bolyai, Chemia, 2021, 66(3).

5.  Udrea, A. M.; Gradisteanu Pircalabioru, G.; Boboc, A. A.; Mares, C.; Dinache, A.; Mernea, M.; Avram, S. “Advanced Bioinformatics Tools in the Pharmacokinetic Profiles of Natural and Synthetic Compounds with Anti-Diabetic Activity”. Biomolecules, 2021, 11(11), 1692, doi.org/10.3390/biom11111692.

 

Papers under review

1. Ispas, C.G.; Raduly, F.M.; Raditoiu, V.; Stan, R.; Raditoiu, A.; Purcar, V. “Extraction and characterization of bioactive compounds from ginger (Zingiber officinale)”, U.P.B. Sci. Bull.

 

Communications

1. Ispas, C.G.; Raduly, F.M.; Raditoiu, V.; Raditoiu, A.; Purcar, V. Extraction by ultrasounds and characterization of bioactive compounds from Zingiber

Officinale, Simpozionul international „Priorities of chemistry for a sustainable development-PRIOCHEM”. XVII th Edition, October 27 – 29, 2021,

Bucuresti, ROMANIA.

2. Ispas, C.G.; Raduly, F.M.; Raditoiu, V.; Stan, R.; Raditoiu, A.; Purcar, V. Textile Coatings Based on Ginger Extracts with Antimicrobial Activity, Virtual

International Scientific Conference on “Applications of Chemistry in Nanosciences and Biomaterials Engineering – NanoBioMat 2021”, 25-27 November

2021, Bucuresti, ROMANIA.

3. M. Raduly, V. Raditoiu, A. Raditoiu, V. Purcar, G. Ispas, Curcumin derived pyrazoles with potential antitumor activities, European Symposium on

Organic Chemistry-ESOC, Virtual mini symposium, 2021.

4. M. Stan, I. C. Nica, C. D. Nițu, M. Raduly, A. Dinischiotu, A. Speranta, D. Mihailescu, M. Mernea, In vitro study of curcumin derivatives with potential antitumor activity, 1st International Electronic Conference on Biomedicine, 2021.

5. M. Raduly, V. Raditoiu, A. Raditoiu, L. Wagner, V. Purcar, G. Ispas, R. Manea, C. A. Nicolae, Synthesis of asymmetric β-diketone derivatives and their complexation with lanthanides, 23rd JCF Frühjahrssymposium-online, 2021.

6. M. Raduly, V. Raditoiu, A. Raditoiu, V. Purcar, G. Ispas, R. Manea, A. Frone, M.L. Jecu, M. Călin, C. A. Nicolae, Curcumin derivatives immobilized in silica

matrices by sol-gel processes used as bioactive coatings, International Congress of „Apollonia” University from Iași, Romania, 2021.

7. Nica, I. C.; Mernea, M.; Stoian, Gh.; Dinischiotu, A. Natural aspirin-like compounds from white willow (Salix alba) bark extract prevent structural changes of human hemoglobin during in vitro non-enzymatic glycation and fructation, preserving its peroxidase and esterase activity, Med. Sci. Forum 2021, 2(1), 23; doi.org/10.3390/CAHD2020-08602.

8. Avram, S., et al. The study of natural compounds as antidepressants by bioinformatics methods, Presented at the 1st International Electronic Conference on Biomedicine. Vol. 1. 2021, https://doi.org/10.3390/ECB2021-10268.

9. Nitu, D. C., et al. Evaluating the ability of some curcumin derivatives to induce oxidative stress in cervix cancer, Presented at the 4th International European Conference on Interdisciplinary Scientific Research, 8-9 august 2021, Warsaw, Poland.

10. M. Stan, I. C. Nica, M. Raduly, M. Mernea, Evaluation of protective effect of some curcumin derivatives against gamma radiations, Al XIV-lea Congres

Național de Citometrie, 24-26 noiembrie 2021.

Other scientific events:

International Exhibition of Inventions – INVENTICA 2021, June 23-25, Iasi, Romania.

Contact

National Institute for Research & Development in Chemistry and Petrochemistry – ICECHIM

202 Splaiul Independentei, 6th District, 060021 Bucharest, Romania

Project manager: Florentina Monica Raduly

Phone: +40 21 316.30.62/118

Fax: +40 21 312.34.93

E-mail:

Web:  www.icechim.ro