MD1824Z

MD1824Z - Use of 4-(2-(biphenyl-4-yl)-2-oxoethyl)-1-((2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl)methyl)-1H-1,2,4-triazole-4-ium bromide as an active compound against Fusarium avenaceum and Fusarium oxysporum fungi

Info

Publication number: MD1824Z
Application number: MDS20240111A
Authority: MD
Inventors: Флюр МАКАЕВ; Галина ЛУПАШКУ; Еуджения СТЫНГАЧ; Сергей ПОГРЕБНОЙ; Наталия СУКМАН; Лучиан ЛУПАШКУ; Светлана ГАВЗЕР; Николае КРИСТЯ
Original assignee: Публичное Учреждение Государственный Университет Молдовы
Priority date: 2023-09-28
Filing date: 2023-09-28
Publication date: 2025-10-31
Also published as: MD1824Y

Abstract

The invention relates to chemistry and agriculture, in particular to the use of a 1,2,4-triazole derivative as a fungicidal agent againstFusarium avenaceumandFusarium oxysporum.According to the invention, claimed is the use of 4-(2-(biphenyl-4-yl)-2-oxoethyl)-1-((2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl)methyl)-1H-1,2,4-triazole-4-ium bromide as an active compound against the fungiF. avenaceumandF. oxysporum- one of the causative agents of root rot.The technical result consists in increasing the fungitoxic activity of the compound according to the invention in relation to the closest analogue by 20…51% forF. avenaceumand by 19…50% forF. oxysporum, in the concentration range of 0.00125…0.01%.

Description

Invenţia se referă la chimie şi agricultură, şi în special la aplicarea unui derivat cuaternar al 1,2,4-triazoluluiîn calitate de compus activ contra fungilorFusarium avenaceumşiFusarium oxysporum. The invention relates to chemistry and agriculture, and in particular to the application of a quaternary derivative of 1,2,4-triazole as an active compound against the fungi Fusarium avenaceum and Fusarium oxysporum.

FungiiF. avenaceumşiF. oxysporumsunt agenţi patogeni ai putregaiului de rădăcină la grâu şi ai maladiilor spicelor la culturile păioase (Lupaşcu G. Putregaiul de rădăcină la grâul comun de toamnă. Chişinău: Print-Caro, 2020, 120 p.). The fungi F. avenaceum and F. oxysporum are pathogens of root rot in wheat and ear diseases in grassy crops (Lupaşcu G. Root rot in common autumn wheat. Chişinău: Print-Caro, 2020, 120 p.).

Conform datelor recente, contaminarea crescută a culturilor de grâu şi orz din Europa şi Asia cu micotoxine emergente, cum ar fi eniatinele sau bovericina, produse deF. avenaceumsugerează că această specie ar putea fi implicată în viitoarele crize de siguranţă alimentară (Ponts N., Gautier Ch., Gouzy J. et al. Evolution ofFusarium tricinctumandFusarium avenaceummitochondrial genomes is driven by mobility of introns and of a new type of palindromic microsatellite repeats. BMC Genomics, BioMed Central, 2020, 21 (1), p. 16 (10.1186/s12864-020-6770-2)). Întrucât speciileFusarium, ca şi multe alte micromicete, uşor se adaptează la preparatele chimice utilizate în măsurile de protecţie a plantelor, sunt deosebit de actuale cercetările cu privire la identificarea noilor compuşi cu activitate antifungică, ceea ce a şi prezentat scopul prezentelor investigaţii. According to recent data, the increased contamination of wheat and barley crops in Europe and Asia with emerging mycotoxins, such as eniatins or bovericin, produced by F. avenaceum suggests that this species could be involved in future food safety crises (Ponts N., Gautier Ch., Gouzy J. et al. Evolution of Fusarium tricinctum and Fusarium avenaceum mitochondrial genomes is driven by mobility of introns and of a new type of palindromic microsatellite repeats. BMC Genomics, BioMed Central, 2020, 21 (1), p. 16 (10.1186/s12864-020-6770-2)). Since Fusarium species, like many other micromycetes, easily adapt to chemical preparations used in plant protection measures, research on the identification of new compounds with antifungal activity is particularly relevant, which is the purpose of the present investigations.

Este cunoscută aplicarea compusului (Z)-1-(2,4-diclorfenil)-5-metil-2-(1H-1,2,4-triazol-1-il)hex-1-en-3-onă în calitate de remediu antifungic către fungiiAlternaria alternata şi Fusarium aquaeductuumcare reprezintă agenţi cauzali frecvent întâlniţi la putregaiul de rădăcină [1]. The compound (Z)-1-(2,4-dichlorophenyl)-5-methyl-2-(1H-1,2,4-triazol-1-yl)hex-1-en-3-one is known to be used as an antifungal agent against the fungi Alternaria alternata and Fusarium aquaeductuum, which are frequently encountered causal agents of root rot [1].

Dezavantajul acestei aplicări constă în aceea că activitatea antifungică nu este la un nivel suficient de înaltă. The disadvantage of this application is that the antifungal activity is not at a sufficiently high level.

Problema rezolvată de invenţie constă în extinderea gamei de aplicări a preparatelor din clasa 1,2,4-triazolilor în agricultură pentru combaterea putregaiului de rădăcină provocat de fungiiF. avenaceumşiF. oxysporum. The problem solved by the invention consists in expanding the range of applications of preparations from the 1,2,4-triazole class in agriculture for combating root rot caused by the fungi F. avenaceum and F. oxysporum.

Esenţa invenţiei constă în aceea că se revendică aplicarea unui compus din clasa 1,2,4 triazolilor:bromură de 4-(2-(bifenil-4-il)-2-oxoetil)-1-((2-(2,4-diclorfenil)-4-propil-1,3-dioxolan-2-il)metil)-1H-1,2,4-triazol-4-iu în calitate de compus activ contra ciupercilor fitopatogene din speciileF. avenaceumşiF. oxysporum -unii dintre agenţii cauzali ai putregaiului de rădăcină. Concentraţiile active variază în diapazonul 0,00125…0,01%. The essence of the invention is that the application of a compound from the class of 1,2,4 triazoles is claimed: 4-(2-(biphenyl-4-yl)-2-oxoethyl)-1-((2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl)methyl)-1H-1,2,4-triazol-4-yl bromide as an active compound against phytopathogenic fungi of the species F. avenaceum and F. oxysporum - some of the causative agents of root rot. The active concentrations vary in the range of 0.00125…0.01%.

Avantajale invenţiei constau în aceea că aplicarea compusului - bromură de 4-(2-(bifenil-4-il)-2-oxoetil)-1-((2-(2,4-diclorfenil)-4-propil-1,3-dioxolan-2-il)metil)-1H-1,2,4-triazol-4-iu contribuie la sporirea activităţii fungitoxice pentru unii dintre agenţii cauzali ai putregaiului de rădăcină- F. avenaceumşiF. oxysporumîn raport cu soluţia cea mai apropiată. The advantages of the invention consist in the fact that the application of the compound - 4-(2-(biphenyl-4-yl)-2-oxoethyl)-1-((2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl)methyl)-1H-1,2,4-triazol-4-yl bromide contributes to the enhancement of the fungitoxic activity for some of the causative agents of root rot - F. avenaceum and F. oxysporum in relation to the closest solution.

Rezultatul tehnic constă în sporirea activităţii fungitoxice a compusului din invenţie în raport cu soluţia cea mai apropiată cu 20…51% pentru fungulF. avenaceumşi cu 19…50% pentruF. oxysporumîn intervalul de concentraţii 0,01….0,00125%, respectiv, în ultimele zile de cultivare a fungilor. The technical result consists in increasing the fungitoxic activity of the compound of the invention in relation to the closest solution by 20…51% for the fungus F. avenaceum and by 19…50% for F. oxysporum in the concentration range 0.01….0.00125%, respectively, in the last days of cultivation of the fungi.

Exemplu de realizare a invenţiei Example of embodiment of the invention

1-((2-(2,4-diclorfenil)-4-propil-1,3-dioxolan-2-il)metil)-1H-1,2,4-triazol(1)şi 1-(bifenil-4-il)-2-bromoetanonă(2)au fost procurate de la firma Aldrich. 1-((2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl)methyl)-1H-1,2,4-triazole (1) and 1-(biphenyl-4-yl)-2-bromoethanone (2) were purchased from Aldrich.

Sinteza compusului(3)decurge conform schemei de realizare: The synthesis of compound (3) proceeds according to the following scheme:

Amestecul reactant alcătuit din părţi echimolare de 1-((2-(2,4-diclorfenil)-4-propil-1,3-dioxolan-2-il)metil)-1H-1,2,4-triazol(1)şi 1-(bifenil-4-il)-2-bromoetanonă(2)în acetonitril, se fierbe timp de 2 ore (control periodic cu ajutorul CSS). Apoi din soluţia reactantă se evaporă excesul de acetonitril în vid. Rămăşiţa solidă se purifică din acetonitril. Substanţa(3)cristalină se filtrează şi se usucă la temperatura camerei. The reaction mixture consisting of equimolar parts of 1-((2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl)methyl)-1H-1,2,4-triazole(1) and 1-(biphenyl-4-yl)-2-bromoethanone(2) in acetonitrile is boiled for 2 hours (periodic control by means of TLC). Then the excess acetonitrile is evaporated from the reaction solution in vacuo. The solid residue is purified from acetonitrile. The crystalline substance (3) is filtered and dried at room temperature.

Randamentul reacţiei este de 86%. The reaction yield is 86%.

Punctul de topire - 127-129ºC. Melting point - 127-129ºC.

Structura compusului(3)este confirmată pe baza analizei elementelor şi datelor spectrale: C29H28BrCl2N3O3; M: 617,36. The structure of compound (3) is confirmed based on elemental analysis and spectral data: C29H28BrCl2N3O3; M: 617.36.

Calculat, (%): C - 56,42; H - 4,57; N - 6,81 Calcd., (%): C - 56.42; H - 4.57; N - 6.81

Stabilit, (%): C - 56,11; H - 4,18; N - 6,42 Determined, (%): C - 56.11; H - 4.18; N - 6.42

IR (ν,cm-1): 3399; 2958; 2931; 1693; 1603; 1578; 1559; 1527; 1486; 1463; 1429; 1404; 1377; 1338; 1238; 1196; 1166; 1105; 1063; 1024; 990; 912; 867; 838; 815; 799; 763; 741; 721; 696. IR (ν,cm-1): 3399; 2958; 2931; 1693; 1603; 1578; 1559; 1527; 1486; 1463; 1429; 1404; 1377; 1338; 1238; 1196; 1166; 1105; 1063; 1024; 990; 912; 867; 838; 815; 799; 763; 741; 721; 696.

Obţinerea unui singur izomer(3)este confirmată prin metoda RMN. The obtaining of a single isomer (3) is confirmed by NMR.

Spectrul RMN1H (400 MHz, DMSO-d6, δ, ppm,J/Hz): 10,19 (s, 1H, H-14), 9,21 (s, 1H, H-15), 8,18 (d,J= 8,5, 2H, H-19), 8,00 (d,J= 8,5, 2H, H-20), 7,83 (d,J= 7,6, 2H, H-23), 7,75 (d,J= 2.1, 1H, H-3), 7,67 (d,J= 8,5, 1H, H-6), 7,58-7,53 (m, 3H, H-24), 7,48 (t,J= 7,3, 1H, H-25), 6,23 (s, 2H, H-16), 5,11 (s, 2H, H-13), 4,01 (t,J= 6,5, 1H, H-8), 3,97 (kvt,J= 6,5, 1H, H-9), 3,44 (dd,J= 8,0, 7,0, 1H, H-8), 1,41-1,19 (m, 4H, H-10,11), 0,87 (t,J= 7,1, 3H, H-12). 1H NMR spectrum (400 MHz, DMSO-d6, δ, ppm,J/Hz): 10.19 (s, 1H, H-14), 9.21 (s, 1H, H-15), 8.18 (d,J= 8.5, 2H, H-19), 8.00 (d,J= 8.5, 2H, H-20), 7.83 (d,J= 7.6, 2H, H-23), 7.75 (d,J= 2.1, 1H, H-3), 7.67 (d,J= 8.5, 1H, H-6), 7.58-7.53 (m, 3H, H-24), 7.48 (t,J= 7.3, 1H, H-25), 6.23 (s, 2H, H-16), 5.11 (s, 2H, H-13), 4.01 (t,J= 6.5, 1H, H-8), 3.97 (kvt,J= 6.5, 1H, H-9), 3.44 (dd,J= 8.0, 7.0, 1H, H-8), 1.41-1.19 (m, 4H, H-10,11), 0.87 (t, J = 7.1, 3H, H-12).

Spectrul RMN13C (101 MHz, DMSO-d6, δ, ppm): 190,40 (C-17), 145,45 (C-15), 145,61(C-16), 138,95 (C-4), 135,44 (C-2), 135,44 (C-2), 134,85 (C-18), 132,99 (C-22),132,69 (C-21), 131,27 (C-3), 130,69 (C-6), 129,67 (C-19), 129,46 (C-20), 129,27 (C-5), 128,03 (C-27), 127,94 (C-1), 127,74 (C-23), 127,58 (C-24), 106,02 (C-7), 76,95 (C-9), 70,29 (C-8), 56,08 (C-13), 54,31 (C-16), 34,18 (C-10), 18,89 (C-11), 14,24 (C-12). 13C NMR spectrum (101 MHz, DMSO-d6, δ, ppm): 190.40 (C-17), 145.45 (C-15), 145.61 (C-16), 138.95 (C-4), 135.44 (C-2), 135.44 (C-2), 134.85 (C-18), 132.99 (C-22),132.69 (C-21), 131.27 (C-3), 130.69 (C-6), 129.67 (C-19), 129.46 (C-20), 129.27 (C-5), 128.03 (C-27), 127.94 (C-1), 127.74 (C-23), 127.58 (C-24), 106.02 (C-7), 76.95 (C-9), 70.29 (C-8), 56.08 (C-13), 54.31 (C-16), 34.18 (C-10), 18.89 (C-11), 14.24 (C-12).

Compusul a fost obţinut conform procedeului descris în cererea de brevet: EP 0026990 A1 1981.04.15 The compound was obtained according to the process described in the patent application: EP 0026990 A1 1981.04.15

Izolarea fungilorF. avenaceumşiF. oxysporum- agenţii cauzali ai putregaiului de rădăcină la grâu s-a efectuat în condiţii aseptice pe mediu PDA (Potatoes Dextrosis Agar) (Методы экспериментальной микологии. Киев: Наукова думка, 1982. 550 p.). Acest mediu este unul din cele mai optime medii pentru izolarea, cultivarea şi cercetarea caracterelor morfologo-culturale ale patogenilor menţionaţi. Au fost utilizate fragmente mici de ţesut cu semne de putrefacţie de la baza tulpinii de grâu. Fragmentele au fost aseptizate în soluţie de hipoclorit de calciu de 2% timp de 1-2 min, după care s-au clătit de 2-3 ori în apă distilată, presat între 2 foiţe de hârtie de filtru şi plasat pe mediu în preajma flăcării de gaz. The isolation of the fungi F. avenaceum and F. oxysporum - the causal agents of root rot in wheat was carried out under aseptic conditions on PDA (Potatoes Dextrose Agar) medium (Methods of experimental mikology. Kiev: Naukova Dumka, 1982. 550 p.). This medium is one of the most optimal media for the isolation, cultivation and research of the morphological and cultural characteristics of the mentioned pathogens. Small fragments of tissue with signs of rotting from the base of the wheat stem were used. The fragments were asepticized in a 2% calcium hypochlorite solution for 1-2 min, after which they were rinsed 2-3 times in distilled water, pressed between 2 sheets of filter paper and placed on the medium near a gas flame.

Identificarea patogenilor s-a efectuat în baza caracteristicelor macro- şi microscopice conform determinatoarelor (Билай В.И. Фузарии. Киев: Наукова думка, 1977, 422 p.; Barnett H.L., Hunter B.B. Illustrated Genera of Imperfect Fungi, fourth edition. APS Press, 1998, 218 p.). Identification of pathogens was carried out based on macro- and microscopic characteristics according to the determinants (Bilay V.I. Fuzarii. Kiev: Naukova Dumka, 1977, 422 p.; Barnett H.L., Hunter B.B. Illustrated Genera of Imperfect Fungi, fourth edition. APS Press, 1998, 218 p.).

Compuşii din invenţie şi soluţia cea mai apropiată au fost suplimentaţi la mediul nutritiv PDA în concentraţiile 0,01 (1); 0,005 (2); 0,0025 (3); 0,00125 (4) care s-a aseptizat prin autoclavare la presiunea de 0,5 atm timp de 30 min. Martorul indicat în tabel reprezintă mediu nutritiv PDA nesuplimentat cu preparat. Mediul aseptizat, fierbinte s-a turnat în cutii Petri, câte 10 mL în fiecare. După solidificarea mediului, fungii au fost însămânţaţi - câte un disc de PDA cu miceliul fungului, cu diametrul de 4 mm în centrul cutiei Petri. Cutiile cu fungii însămânţaţi, au fost menţinute în termostat la temperatura de 24°C. Înregistrarea diametrului coloniilor (câte 2 diametre perpendiculare, media cărora a servit ca indice biometric) s-a efectuat din ziua 3-4 de la însămânţare, în dependenţă de viteza de creştere a fungilor. Experienţele s-au efectuat în 5 repetiţii. Datele au fost prelucrate statistic în pachetul de soft STATISTICA 7. The compounds of the invention and the closest solution were supplemented to the PDA nutrient medium in concentrations of 0.01 (1); 0.005 (2); 0.0025 (3); 0.00125 (4) which was sterilized by autoclaving at a pressure of 0.5 atm for 30 min. The control indicated in the table represents PDA nutrient medium not supplemented with the preparation. The sterilized, hot medium was poured into Petri dishes, 10 mL each. After the medium solidified, the fungi were seeded - one PDA disk with the fungal mycelium, with a diameter of 4 mm in the center of the Petri dish. The boxes with the seeded fungi were kept in a thermostat at a temperature of 24°C. The colony diameter was recorded (2 perpendicular diameters, the average of which served as a biometric index) from day 3-4 after seeding, depending on the growth rate of the fungi. The experiments were performed in 5 repetitions. The data were statistically processed in the STATISTICA 7 software package.

Tabelul 1 Table 1

Influenţa derivaţilor triazolici asupra creşterii funguluiF. avenaceum Influence of triazole derivatives on the growth of the fungus F. avenaceum

Variantă Concen-traţie, % Ziua 3 Ziua 4 Ziua 5 Diametrul coloniei, mm % din martor Diametrul coloniei, mm % din martor Diametrul coloniei, mm % din martor Martor - 51,6±1,6 - 79,4±1,6 - 90,0±0 - Invenţia 0,01% 8,3±0,7* 16,0 12,9±1,6* 16,2 18,4±1,0* 20,4 0,005 10,6±0,4* 20,6 17,9±0,7* 22,5 22,4±0,1* 24,9 0,0025 13,1±1,9* 25,5 22,0±2,0* 27,7 29,0±1,7* 32,2 0,00125 15,6±1,5* 30,3 24,0±0,9* 30,2 31,9±0,2* 35,4 Soluţia cea mai apropiată 0,01% 15,3±1,6* 29,6 24,4±1,8* 30,7 36,1±2,5* 40,1 0,005 20,4±0,4* 39,5 34,5±0,6* 43,5 50,4±0,2* 56,0 0,0025 26,8±1,2* 51,9 47,3±1,1* 59,5 68,0±1,4* 75,6 0,00125 32,8±1,6* 63,5 53,9±1,4* 67,9 77,5±1,6* 86,1Variant Concentration, % Day 3 Day 4 Day 5 Colony diameter, mm % of control Colony diameter, mm % of control Colony diameter, mm % of control Control - 51.6±1.6 - 79.4±1.6 - 90.0±0 - Invention 0.01% 8.3±0.7* 16.0 12.9±1.6* 16.2 18.4±1.0* 20.4 0.005 10.6±0.4* 20.6 17.9±0.7* 22.5 22.4±0.1* 24.9 0.0025 13.1±1.9* 25.5 22.0±2.0* 27.7 29.0±1.7* 32.2 0.00125 15.6±1.5* 30.3 24.0±0.9* 30.2 31.9±0.2* 35.4 Closest solution 0.01% 15.3±1.6* 29.6 24.4±1.8* 30.7 36.1±2.5* 40.1 0.005 20.4±0.4* 39.5 34.5±0.6* 43.5 50.4±0.2* 56.0 0.0025 26.8±1.2* 51.9 47.3±1.1* 59.5 68.0±1.4* 75.6 0.00125 32.8±1.6* 63.5 53.9±1.4* 67.9 77.5±1.6* 86.1

*- diferenţă de martor statistic semnificativă, p<0,05. *- statistically significant difference from control, p<0.05.

Conform rezultatelor prezentate, se atestă sporirea activităţii fungitoxice în ultima zi de cultivare în urma aplicării compusului din invenţie în raport cu cel din cadrul soluţiei celei mai apropiate asupraF. avenaceumcu 20…51% în intervalul de concentraţii 0,01….0,00125%, respectiv. According to the presented results, the fungitoxic activity increased on the last day of cultivation following the application of the compound of the invention compared to that of the closest solution on F. avenaceum by 20…51% in the concentration range 0.01….0.00125%, respectively.

Tabelul 2 Table 2

Influenţa derivaţilor triazolici asupra creşterii funguluiF.oxysporum The influence of triazole derivatives on the growth of the fungus F.oxysporum

Variantă Concentraţie, % Ziua 6 Ziua 7 Diametrul coloniei, mm % din martor Diametrul coloniei, mm % din martor Martor - 58,9±1,9 - 66,5±2,4 - Invenţia 0,01% 14,0±0,5* 23,8 15,6±0,7* 23,5 0,005 14,8±1,9* 25,0 16,5±1,9* 24,8 0,0025 14,8±0,3* 25,0 16,5±0,7* 24,8 0,00125 16,6±0,6* 28,2 19,3±0,6* 29,0 Soluţia cea mai apropiată 0,01% 22,3±3,0* 37,8 28,3±1,6* 42,5 0,005 32,0±1,0* 54,3 37,6±1,1* 56,6 0,0025 40,5±0,5* 68,7 46,9±0,5* 70,5 0,00125 44,3±0,9* 75,1 52,5±1,9* 79,0Variant Concentration, % Day 6 Day 7 Colony diameter, mm % of control Colony diameter, mm % of control Control - 58.9±1.9 - 66.5±2.4 - Invention 0.01% 14.0±0.5* 23.8 15.6±0.7* 23.5 0.005 14.8±1.9* 25.0 16.5±1.9* 24.8 0.0025 14.8±0.3* 25.0 16.5±0.7* 24.8 0.00125 16.6±0.6* 28.2 19.3±0.6* 29.0 Closest solution 0.01% 22.3±3.0* 37.8 28.3±1.6* 42.5 0.005 32.0±1.0* 54.3 37.6±1.1* 56.6 0.0025 40.5±0.5* 68.7 46.9±0.5* 70.5 0.00125 44.3±0.9* 75.1 52.5±1.9* 79.0

Conform rezultatelor prezentate, se atestă sporirea activităţii fungitoxice în ultima zi de cultivare în urma aplicării compusului din invenţie în raport cu cel din cadrul soluţiei celei mai apropiate asupraF. oxysporumcu 19…50% în intervalul de concentraţii 0,01….0,00125%, respectiv. According to the presented results, the fungitoxic activity increased on the last day of cultivation following the application of the compound of the invention compared to that of the closest solution on F. oxysporum by 19…50% in the concentration range 0.01….0.00125%, respectively.

1. MD 4823 B1 2022.09.30 1. MD 4823 B1 2022.09.30

Claims

Application of 4-(2-(biphenyl-4-yl)-2-oxoethyl)-1-((2-(2,4-dichlorophenyl)-4-propyl-1,3-dioxolan-2-yl)methyl)-1H-1,2,4-triazol-4-yl bromide as an active compound against the fungi Fusarium avenaceum and Fusarium oxysporum.