Project objectives

1) Investigation of the interaction of N-vinylsulfonylpyrrolidine derivatives with amines; determination of the influence of reaction conditions (solvent, temperature) and reagent structure on the synthetic outcome. Synthesis of water-soluble 1-sulfonylpyrrolidines.

2) Investigation of the interaction of N-vinylsulfonylpyrrolidine derivatives with amino acids; determination of the influence of reaction conditions (solvent, temperature) and reagent structure on the synthetic outcome. Synthesis of 1-sulfonylpyrrolidines containing amino acid residues in their structure.

3) Investigation of the reaction of 3-arylidene-1-pyrroline with fluoroquinolone drugs. Determination of the structure of the reaction products using a set of physical analytical methods.

4) Investigation of the reactions of phosphonium and pyridinium salts of 1-sulfonylpyrrolidine with various nucleophiles. Synthesis of new pyrrolidine derivatives that are difficult to obtain by other known methods.

5) Investigation of the cytotoxicity of the obtained compounds toward tumor and normal human cell lines.

Expected results

Synthesis of the starting N-(4,4-diethoxybutyl)ethenesulfonamide and N-vinylsulfonylpyrrolidines according to our previously developed methodology.

Using the most accessible and easily synthesized reagents as examples, the reaction conditions will be optimized to achieve the highest yield of pyrrolidine derivatives. The solvent, temperature, reaction time, and other parameters will be varied.

The interaction of N-vinylsulfonylpyrrolidines with a variety of amines and amino acids, including proteinogenic ones, will be carried out. The dependence of product yield on the substituent in the pyrrolidine ring, as well as on the amino acid used, will be established.

The structural features of the obtained 1-sulfonylpyrrolidines will be studied using a set of physicochemical methods of analysis (¹H, ³¹P, ¹³C NMR spectroscopy, IR spectroscopy, mass spectrometry, and X-ray crystallographic analysis).

Cytotoxicity toward normal and cancer human cell lines will be studied. Acute toxicity in mice will be determined. LD₅₀, LD₁₀₀, and MTD values will be established.

Based on the results of the conducted research, three articles will be published in high-impact journals, and presentations will be delivered at relevant scientific conferences.

1) The results obtained during the course of the project will be published in (three) scientific papers in peer-reviewed journals relevant to the project’s research area:

ChemMedChem, a peer-reviewed scientific journal focused on medicinal chemistry. CiteScore (Scopus) – 5.6. Journal Citation Indicator (Clarivate) – 0.82. Q3. Journal Impact Factor (Clarivate): 3.4 (https://www.scimagojr.com/journalsearch.php?q=4400151608&tip=sid&clean=0);

Journal of Organic Chemistry, a peer-reviewed scientific journal in the field of chemistry. CiteScore (Scopus) – 6.8. Journal Citation Indicator (Clarivate) – 1.1. Q1. Journal Impact Factor (Clarivate): 3.6 https://www.scimagojr.com/journalsearch.php?q=25896&tip=sid&clean=0;

Arabian Journal of Chemistry, an open-access peer-reviewed journal in the field of chemistry. CiteScore (Scopus) – 11. Q2. Journal Impact Factor – 6. Journal Citation Indicator (Clarivate) – 0.95

(https://www.scimagojr.com/journalsearch.php?q=19400158709&tip=sid&clean=0),

as well as in other journals indexed in the Science Citation Index Expanded and ranked in the 1st, 2nd, and/or 3rd quartiles by impact factor in the Web of Science database and/or having a CiteScore percentile of at least 60 in the Scopus database.

In addition, one article or review will be published in the Eurasian Journal of Chemistry (formerly Bulletin of the Karaganda University. Chemistry series) and in other peer-reviewed international or national journals recommended by the Committee for Quality Assurance in Science and Higher Education (CQASHE).

Dissemination of the research results within the scientific community is planned through participation in relevant scientific conferences of various levels.

The publication of monographs, books, and/or book chapters in foreign and/or domestic publishing houses is not planned.

Obtaining patents in foreign patent offices (European, American, Japanese), as well as in domestic or Eurasian patent offices: a patent of the Republic of Kazakhstan or the Russian Federation for an invention will be obtained.

The development of scientific, technical, and engineering documentation is not planned.

Dissemination of the research results among potential users, the scientific community, and the public: dissemination will be carried out through participation in international scientific conferences, as well as through national and regional mass media and social networks.

Other measurable results in accordance with the requirements of the funding call documentation and the specifics of the project. Additional information is provided in this section:

1. Field of application and target consumers of each of the expected results:

   As a result of the implementation of the proposed project, an extensive library of previously undescribed 1-sulfonylpyrrolidines exhibiting high cytotoxicity toward human cancer cell lines will be obtained. A structure–activity relationship analysis will be conducted, and lead compounds demonstrating high selectivity and low toxicity toward normal cells will be identified. This will serve as a basis for the further development of new types of antitumor agents and will contribute to the advancement of medicinal chemistry. Moreover, it will provide a reliable foundation for the practical implementation of new types of highly selective and low-toxicity antitumor agents.

2. Impact of the expected results on the development of the main scientific field and related areas of science and technology:

   Pharmaceutical companies may become the primary target consumers of the project’s scientific results, as the obtained compounds are expected to exhibit a broad spectrum of biological activity, reduced toxicity, and the potential to compete with expensive foreign analogues.

3. Applicability and/or potential for commercialization of the obtained scientific results:

   The obtained scientific results may be applied in the pharmaceutical industry and medicine as antitumor agents, as well as in the educational process for improving curricula, developing laboratory work, and designing practical training for students at various levels.

4. Social, economic, environmental, scientific, technical, multiplicative, and/or other effects of the project results, with justification, including addressing existing regional challenges:

   The results obtained during this project will significantly expand current knowledge in the field of medicinal chemistry.

5. Other direct and indirect results of the project, including their qualitative and quantitative characteristics:

   The results are planned to be presented at international and national conferences.

Results

The synthesis of N-(4,4-diethoxybutyl)ethenesulfonamide was carried out via the reaction of 4,4-diethoxybutan-1-amine with 2-chloroethanesulfonyl chloride in dichloromethane in the presence of triethylamine at room temperature. It is noteworthy that during the reaction, the sulfonyl group is simultaneously introduced at the nitrogen atom and hydrogen chloride is eliminated, resulting in the formation of a double bond.

The reliability of the obtained results is confirmed by the use of modern physicochemical methods: mass spectrometry (MALDI-TOF and ESI), IR spectroscopy, ¹H, ¹³C, and ³¹P NMR spectroscopy, as well as elemental and X-ray crystallographic analysis.

¹H, ¹³C, and ³¹P NMR spectra were recorded on Bruker AVANCE II-400 spectrometers operating at 400.1 MHz (¹H) and 100.6 MHz (¹³C), and on a Bruker Avance-600 spectrometer operating at 600.1 MHz (¹H) and 150.9 MHz (¹³C), referenced to the signals of residual protons of the deuterated solvent or carbon nuclei of the deuterated solvent (DMSO-d₆).

IR spectra were recorded on a Bruker Vector 22 Fourier-transform spectrometer in the range of 400–4000 cm⁻¹. The samples were analyzed as KBr pellets.

Melting points were determined using a Boetius apparatus. Elemental analysis was performed on a Carlo Erba EA 1108 instrument. The completeness of the reactions and the purity of the synthesized compounds were monitored by thin-layer chromatography (TLC) on SORBFIL PTSKh-AF-A-UV plates (Sorbpolymer, Krasnodar, Russia), using a benzene-ethanol (10:1) eluent and UV light as the visualizing agent.

N-vinylsulfonylpyrrolidines containing 1-phenylpyrazolidin-3-one, antipyrine, and diphenylphosphine oxide fragments at the second position of the heterocycle were obtained.

These compounds were obtained as a result of the reaction of N-(4,4-diethoxybutyl)ethenesulfonamide with 1-phenylpyrazolidin-3-one and antipyrine in benzene in the presence of an equimolar amount of trifluoroacetic acid.

The structure of the obtained compounds was confirmed by ¹H and ¹³C NMR spectroscopy and IR spectroscopy data. The formation of the product is clearly supported by the ¹H NMR spectra. In contrast to the spectrum of the initial acetal, the spectrum of the product lacks the quartet and triplet signals corresponding to the ethoxy groups. The presence of the pyrrolidine ring is evidenced by the increased complexity of the ¹H NMR spectrum compared to the starting compound. Specifically, the five protons of the pyrrolidine ring appear as four multiplets, which is associated with the increased complexity of the spin system.

The presence of the vinyl fragment is confirmed by the signals of the methylene group protons in the form of two doublets with chemical shifts of 5.86 ppm (J = 10.0 Hz) and 6.08 ppm (J = 16.6 Hz), as well as the methine proton appearing as a doublet of doublets at 6.36 ppm (J = 16.6 Hz, J = 9.9 Hz).

2-Hetarylpyrrolidines were isolated in yields of 67% and 81%, respectively.

A phosphorylated pyrrolidine containing a vinyl fragment was obtained via the reaction of N-(4,4-diethoxybutyl)ethenesulfonamide with diphenylchlorophosphine in chloroform in the presence of acetic acid. The new product was isolated in 87% yield. The spectral characteristics of this compound are similar to those described above. The presence of the phosphine oxide fragment in this compound was confirmed by ³¹P NMR spectroscopy data. In the ³¹P NMR spectrum of the obtained compound, a singlet with a chemical shift of 31 ppm is observed.

The optimal reaction conditions providing the highest product yield were identified.

For optimization of the process conditions, a model reaction between N-vinylsulfonylpyrrolidine containing a 1-phenylpyrazolidin-3-one fragment and the most accessible amino acid, glycine, was selected. Carrying out the reaction in a boiling EtOH:H₂O (4:1) mixture in the presence of Et₃N for 6 hours afforded the target product in only 15% yield. Increasing the reaction time to 24 hours led to a slight increase in yield. The use of water as a solvent also did not improve the yield. Subsequently, a screening of various basic catalysts was performed. The use of DMAP or pyridine increased the yield to 35%. Unexpectedly, replacing ethanol with methanol and conducting the reaction under reflux for 1 hour resulted in a product yield of 75%. Extending the reaction time to 24 hours allowed the isolation of the pyrrolidine derivative in 93% yield; therefore, these conditions were considered optimal.

The structure of the obtained compound was unambiguously confirmed by ¹H NMR spectroscopy. As a result of the aza-Michael reaction, which proceeds via the vinyl fragment of 2-arylpyrrolidine and the amino group of glycine, a secondary amine is formed. In the ¹H NMR spectrum of the product, the signals of the vinyl fragment in the region of 6 ppm disappear, while two multiplets corresponding to methylene protons appear in the region of 3 ppm.

Based on the results of the conducted studies, one article has been submitted to a journal indexed in the Science Citation Index Expanded, classified as Q2 by impact factor in the Web of Science database, and having a CiteScore percentile of 69 in the Scopus database.

Andrey Smolobochkin, Tanzilya Rizbayeva, Rakhymzhan Turmanov, Almir Gazizov, Nurgali Akylbekov, Saltanat Nakypova, Rakhmetulla Zhapparbergenov, Roza Narmanova, Saltanat Ibadullayeva, Alena Zalaltdinova, Marat Syzdykbayev, Julia Voronina, Anna Lyubina, Alexandra Voloshina, Elena Klimanova, Tatiana Sashenkova, Denis Mishchenko, Alexander Burilov / Design, Synthesis and Biological Evaluation of Taurine Derivatives Containing a Pyrrolidine Moiety, as Potential Anticancer Agents // Journal of Chemistry. Статус статьи: Under Review.