CS276184B6 - Algicidal composition of the N-ethyl-2-pyrrolidinylmethyl ester type of alkoxycarbanilic acids - Google Patents

Algicidal composition of the N-ethyl-2-pyrrolidinylmethyl ester type of alkoxycarbanilic acids Download PDF

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CS276184B6
CS276184B6 CS902338A CS233890A CS276184B6 CS 276184 B6 CS276184 B6 CS 276184B6 CS 902338 A CS902338 A CS 902338A CS 233890 A CS233890 A CS 233890A CS 276184 B6 CS276184 B6 CS 276184B6
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ethyl
kak
alkoxycarbanilic
acids
compounds
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CS902338A
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CS233890A3 (en
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Katarina Rndr Csc Kralova
Frantisek Ing Csc Sersen
Jozef Rndr Csc Csollei
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Univ Komenskeho
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Abstract

Algicídny prostriedok, ktorý ako účinnú látku obsahuje zlúčeninu obecného vzorca I, kde OR znamená alkoxyl so 4 až 7 atómami uhlíka v orto- alebo meta-polohe benzénového jadra. Syntéza týchto zlúčenín je predmetom čs. autorského osvedčenia č. 211247 autorov: Csollei J., Beneš L., Borovanský A., Švec P. (1985). Zlúčeniny podlá vynálezu_posobia_|lgicídne pri končentráciách 1.10 mól.dm a vyšších (tj. pri koncentráciách 3,9 mg.dm a vyšších).Algicidal agent, which contains as an active substance a compound of the general formula I, where OR means alkoxy with 4 to 7 carbon atoms in the ortho- or meta-position of the benzene ring. The synthesis of these compounds is the subject of Czechoslovak author's certificate No. 211247 of the authors: Csollei J., Beneš L., Borovanský A., Švec P. (1985). The compounds according to the invention act_algicidally at concentrations of 1.10 mol.dm and higher (i.e. at concentrations of 3.9 mg.dm and higher).

Description

'-x CS 276184 B6

Vynález sa týká algicídneho prostriedku, ktorý ako účinnú látku obsahuje zlúčeninuobecného vzorca I

kde OR znamená alkoxyl so 4 až 7 atómami uhlíka v orto- alebo meta- polohe benzénovéhojadra.

Syntéza zlúčenín obecného vzorca I je predmetom čs. autorského osvedčenia č. 211247autorov: Csóllei J., Beneš L., Borovanský A., Švec P. (1985). Tieto zlůčeniny vykazujú lo-kálně anestetickú účinnost (Csollei J.: Farm. obzor 50, 407, 1981).

Pri dalšom štúdiu biologickej účinnosti zlúčenín obecného vzorca I sme zistili, žetieto výrazné inhibujú rast a syntézu chlorofylu rias.

Inhibičná účinnost zlúčenín obecného vzorca I na riasy sa testovala stacionárnou kul-tiváciou synchronněj kultúry rias Chlorella vulgaris. Hodnotil sa nárast buniek rias, akoaj obsah syntetizovaného chlorofylu. Podrobný popis metody hodnotenia je uvedený v příkla-de 1. Miera antialgálnej účinnosti zlúčenín obecného vzorca I je vyjádřená údajmi v tab. 1.Inhibičný vplyv na riasy sa sledoval u nasledujúcich zlúčenín obecného vzorca I: 1. N-etyl-2-pyrolidinylmetylester 2-butoxykarbanilovej kyseliny, hydrochlorid (pracovnéoznačenie KAK 2224) 2. N-ety1-2-pyrolidinylmetylester 2-pentyloxykarbanilovej kyseliny, hydrochlorid (KAK 2225) 3. N-etyl-2-pyrolidinylmetylester 2-hexyloxykarbanilovej kyseliny, hydrochlorid (KAK 2226) 4. N-etyl-2-pyrolidinylmetylester 2-heptyloxykarbanilovej kyseliny, hydrochlorid (KAK 2227) 5. N-etyl-2-pyrolidinylmetylester 3-butoxykarbanilovej kyseliny, hydrochlorid (KAK 2234) 6. N-etyl-2-pyrolidinylmetylester 3-pentyloxykarbanilovej kyseliny, hydrochlorid (KAK 2235) 7. N-etyl-2-pyrolidinylmetylester 3-hexyloxykarbanilovej kyseliny, hydrochlorid (KAK 2236) 8. N-cty1-2-pyrolidinylmctylcstcr 3-hcptyloxykarbanilovcj kyseliny (KAK 2237). TABULKA 1

Inhibičná účinnost látok obecného vzorca I na riasy Chlorella vulgaris Látka IC50 MIC IC50 10-6mól.dm“3 syntézy syntézy rastu chlorofylu chlorofylu rias KAK 2224 39,4 66,1 53,7 KAK 2225 23,4 60,3 30,2 KAK 2226 14,5 50,1 20,9 KAK 2227 8,0 15,8 12,0 KAK 2234 13,2 39,8 20,9 KAK 2235 12,3 25,1 15,0 KAK 2236 6,8 10,0 8,7 KAK 2237 6,9 11,0 8,3 CS 276184 B6 IC,-0 značí koncentráciu študovanej látky, pri ktorej dochádza k 50 %nej inhibícii hodnotc-ncho parametra rias vzhladom na kontrolné vzorky; MIC značí miniraálnu koncentráciu študovanej látky, pri ktorej dochádza k úplncj inhibíciihodnotenóho parametra rias.

Nasledujúci příklad bližšie osvětluje inhibičnú účinnost zlúčenín podlá vynálezu nazelené riasy. Příklad 1 Účinok zlúčenín podlá vynálezu na rast rias Chlorella vulgaris a obsahsyntetizovaného chlorofylu

Riasy Chlorella vulgaris sa pěstovali stacionárnou kultiváciou pri teplote 25+1 'c, pri svetelnom režime 16 hodin světlo - 8 hodin tma, v kultivačnom médiu podlá Setlíka (Setlík I.: Annual Report of Algolog Lab. for 1967, 71-140, Třeboň, 1968). K živnému médiu sa přidával roztok študovanej zlúčeniny v destilovanéj vodě takej koncentrácie, aby sa získala séria kultivačných baniek s bdstupňovanou koncentráciou látky a odstupňovaným bio-·· * logickým účinkom. Banky sa inokulovali přesným objemom homogénnej suspenzie rias. Po7-dennej kultivácii sa objem kultivačného média doplnil destilovanou vodou na povodný ob-jem (vyrovnanie odparu vody) a v suspenzii buniek sa merala absorbencia pri 660 nm (úměr-ná množstvu buniek rias) a obsah chlorofylu po jeho extrakcii dimetylformamidom z odcen-trifugovaných buniek rias. Celkový obsah chlorofylu rias v kultivačnom médiu sa vyhodno-til podlá rovnice: Chl /mg.dm 3/ = 17,9 Ag^? + 8,08 Agg^ g, kde A je absorbancia vzorkypri 647, resp. 664,5 nm v 1 cm kývete (Inskeep W. P., Bloom P.R.: Plant Physiol. 77 (2),483-5, 1985). Získané hodnoty týchto parametrov sa porovnávali s hodnotami parametrov kontrolnýchvzoriek (vyhodnotené ako % kontroly). Na základe týchto hodnot sa extrapoláciou vyčíslilikoncentrácie študovaných zlúčenín, pri ktorých sa dosiahla 50 %ná inhibícia (ICgg) a vpřípade hodnot chlorofylu aj úplná inhibícia (MIC). Z hodnot absorbancie sa MIC nevyhodno-covala, nakolko touto metodou sa stanovujú tiež odfarbené mřtve buňky rias, a teda aj priúplnéj inhibícii sa namerajú určité hodnoty absorbancie.

Například v případe testovania účinku zlúčeniny KAK 2237 sa k 12 ml kultivačného mé-dia s inokulom rias přidalo 0,12 ml vodného roztoku zlúčeniny KAK 2237 nasledovných kon-centrácií: 2,5.10-3, 1.10-3, 7,5.1θ"4, 5.10-4 a 1.10-4 mól.dra~3. Skutočné koncentráciezlúčeniny KAK 2237 v kultivačnom médiu a příslušné hodnoty inhibície rastu rias a synté-’zy chlorofylu, vyjádřené v % kontrolných vzoriek sú uvedené v tab. 2.

Hodnoty koncentrácie študovanej zlúčeniny, pri ktorej dochádza k 50 %nej, resp. úpl-nej inhibícii študovaného parametra rias sa zistuje extrapoláciou zo závislosti vyššieuvedených hodnot % zistenej inhibície sledovaného parametra od logaritmu aplikovanej kon-centrácie študovanej zlúčeniny. TABULKA 2

Inhibícia rastu rias a syntézy chlorofylu v přítomnosti zlúčeniny KAK 2237 G"' = smerodajná odchýlka

Konc. zlúč. KAK 2237 % kontroly + ”*6 ""3 10 mól.dm Chlorofyl Absorbancia 25 0 12,5 + 0 10 9,3 + 0,7 18,5 + 2,0 7,5 47,5 + 7,7 63,3 + 1,1 5,0 85,4 + 3,6 86,0 + 3,0 1,0 110,6 + 3,0 106,2 + 4,4

CS-276184 B6

The present invention relates to an algicidal composition which comprises a compound of formula I as active ingredient

wherein OR is 4 to 7 carbon atoms in the ortho or meta position of the benzene ring.

The synthesis of compounds of formula I is the subject of MS. Certificate No. 211247Author: Csóllei J., Beneš L., Borovanský A., Švec P. (1985). These compounds exhibit locally anesthetic activity (Csollei J .: Farming Horizon 50, 407, 1981).

In a further study of the biological activity of the compounds of formula I, we have found that these significantly inhibit the growth and synthesis of algal chlorophyll.

The algae inhibitory activity of the compounds of formula I was tested by stationary culture of synchronous algal culture Chlorella vulgaris. An increase in algal cells was evaluated, such as the content of chlorophyll synthesized. A detailed description of the evaluation method is given in Example 1. The level of anti-oral efficacy of the compounds of Formula I is shown in Table 1 below. 1. The algae inhibitory effect was observed for the following compounds of the formula I: 1. N-ethyl-2-pyrrolidinylmethyl ester of 2-butoxycarbanilic acid hydrochloride (KAK 2224 working label) 2. N-ethyl-2-pyrrolidinylmethyl ester of 2-pentyloxycarbanilic acid hydrochloride (KAK 2225) 3. N-Ethyl-2-pyrrolidinylmethyl ester of 2-hexyloxycarbanilic acid hydrochloride (KAK 2226) 4. N-Ethyl-2-pyrrolidinylmethyl ester of 2-heptyloxycarbanilic acid hydrochloride (KAK 2227) 5. N-ethyl-2- 3-butoxycarbanilic acid pyrrolidinylmethyl ester hydrochloride (KAK 2234) 6. 3-pentyloxycarbanilic acid N-ethyl-2-pyrrolidinylmethyl ester hydrochloride (KAK 2235) 7. N-ethyl-2-pyrrolidinylmethyl ester of 3-hexyloxycarbanilic acid hydrochloride (KAK 2236) N-ethyl-2-pyrrolidinylmethylcyclohexylcarbonic acid (KAK 2237). TABLE 1

Inhibitory activity of compounds of formula I on Chlorella vulgaris algae Substance IC50 MIC IC50 10-6 mole.dm 3 chlorophyll chlorophyll synthesis synthesis algae KAK 2224 39,4 66,1 53,7 KAK 2225 23,4 60,3 30,2 KAK 2226 14,5 50,1 20,9 KAK 2227 8,0 15,8 12,0 KAK 2234 13,2 39,8 20,9 KAK 2235 12,3 25,1 15,0 KAK 2236 6,8 10 IC, -0 indicates the concentration of the study substance at which 50% inhibition of the algal parameter is observed with respect to the control samples; MIC denotes the miniraal concentration of the study substance at which complete inhibition of the measured algal parameter occurs.

The following example further illustrates the green algae inhibitory activity of the compounds of the invention. Example 1 Effect of the compounds of the invention on the growth of algae Chlorella vulgaris and the content of synthesized chlorophyll

Algae Chlorella vulgaris were grown by stationary cultivation at 25 + 1 ° C, under light regime 16 hours light - 8 hours dark, in culture medium by Setlík (Setlík I .: Annual Report of Algolog Lab. For 1967, 71-140, Třeboň , 1968). To the nutrient medium, a solution of the study compound in distilled water was added to a concentration to obtain a series of culture flasks with a graded concentration of substance and a graded bio-logical effect. The flasks were inoculated with an exact volume of homogeneous algae suspension. After 7 days of culture, the volume of the culture medium was supplemented with distilled water to the flood volume (water vapor equalization) and the absorbance at 660 nm (proportional to the amount of algal cells) and the chlorophyll content after extraction with dimethylformamide from the effluent were measured in the cell suspension. cell algae. The total algal chlorophyll content in the culture medium was evaluated according to the equation: Chl / mg.dm 3 / = 17.9 Ag =? + 8.08 Agg / g, where A is the absorbance of the sample at 647, respectively. 664.5 nm in 1 cm nod (Inskeep WP, Bloom PR: Plant Physiol. 77 (2), 483-5, 1985). The obtained values of these parameters were compared with the values of the control sample parameters (evaluated as% of control). Based on these values, extrapolation was used to quantify the concentration of the compounds studied in which 50% inhibition (ICgg) and complete inhibition (MIC) were achieved. From the absorbance values, the MIC was not evaluated because this method also determined bleached algal cell counts, and thus, some absorbance values were also measured by total inhibition.

For example, when testing the effect of compound KAK 2237, 0.12 ml of an aqueous solution of KAK 2237 of the following concentrations was added to 12 ml of algal inoculum culture medium: 2.5.10-3, 1.10-3, 7.5.1θ "4 The actual concentrations of the compound KAK 2237 in the culture medium and the respective algal growth inhibition and chlorophyll synthesis values, expressed as a percentage of the control samples, are shown in Table 2.

The concentration of the study compound at which 50% of the compound is studied, respectively. the complete inhibition of the algal parameter studied is determined by extrapolation from the dependence of the aforementioned% found inhibition of the observed parameter on the logarithm of the applied compound concentration. TABLE 2

Inhibition of algal growth and chlorophyll synthesis in the presence of KAK 2237 G "" = standard deviation

Conc. bile. KAK 2237% control + ”* 6""3 10 mol.dm Chlorophyll Absorbance 25 0 12.5 + 0 10 9.3 + 0.7 18.5 + 2.0 7.5 47.5 + 7.7 63 , 3 + 1.1 5.0 85.4 + 3.6 86.0 + 3.0 1.0 110.6 + 3.0 106.2 + 4.4

Claims (1)

-·*·¥ CS 276184 B6 PATENTOVÉ NÁROKY Algicídny prostriedok, vyznačený tým, že ako účinnú látku obsahuje zlúčeninu obecnéhovzorca I o -CEg-O-C-EH· OR Í2h5 /1/ kde OR znamená alkoxyl so 4 až 7 atómami uhlíka v orto- alebo meta- polohe benzénovéhojadra.CS 276184 B6 PATENT REQUIREMENTS Algicidal composition, characterized in that it contains as active substance a compound of general formula I o -CEg-OC-EH · OR 2h 5/1 / wherein OR is a C 4 -C 7 alkoxyl in ortho or meta position of the benzene core.
CS902338A 1990-05-14 1990-05-14 Algicidal composition of the N-ethyl-2-pyrrolidinylmethyl ester type of alkoxycarbanilic acids CS276184B6 (en)

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