CS252502B1 - 4-x-3-benzyloxycarbonylmethyl-2-oxobenzothiazolines and method of their preparation - Google Patents

Description

The invention relates to 4-X-3-benzyloxycarbonylmethyl-2-oxobenzothiazolines of the general formula I rot

Ύ = 0 _Jm-ch ₂ cooch.

(<) where

X represents hydrogen, chlorine and a process for their preparation. 3-Benzyloxycarbonylmethyl-2-oxobenzothiazoline and 4-chloro-3-benzyloxycarbonylmethyl-2-oxobenaothiazoline are effective for the dedifferential transformation of the plant organism.

The present syntheses of variously substituted 2-oxobenzothiazolines were mainly oriented on their practical use in the field of pesticides. They have found application as growth regulators (D'Amico JJ: US Pat. 4,371,388 (1893); Chem. Abstract 99,22454 (1983); D'Amico JJ: US Pat. 4171213 (1979); Chem. Abstract 92 58764 (1983);

D. Amico J. J .: US Pat. 4187 097 (1977), Eur. pat. 2613 (1979). Chem. abstr. 92

419,332 (1983); Ueda I .: jap. pat. 92956 (1979); Chem. abstr. 92, 94383 (1983); Jap. pat. 105,605 (1980); Chem. abstr. 93, 216, 681 (1983), of the formula n-3-methyl (ethyl) -2-oxobenzothiazolines (Jap. Pat. 9040 546 (1977); USSR Pat. 713 523 (1978); D'Amico, JJ Europ. U.S. Patent No. 7,772 (1980), Chem Abstract 93, 150241 (1983), Japanese Patent No. 7 9032 787 (1977), USSR Patent No. 668,567 (1978), British Patent No. 1564,182 (1978) NSR U.S. Pat. No. 2846,980 (1978), also as a means to increase the sugar content of sugar-producing plants (D'Amico JJ: U.S. Pat. No. 576,512 (1975)).

R = CH ₃ CN, (CH ₂ ) 5 CN,

CH2COOCH3.

CH (COOCH3) 2,

CH (COO, C ₂ H 5) 2,

C-H2COOH,

CH2CONH2 and also as a herbicide [Sohler E; Seidler L .; Vano J .; Splhacek R .; Fedor E .: Csl. pat. 172,118 (1974)].

The 4-X-3-benzyloxycarbonylmethyl-2-oxobenzothiazolines of the invention have not been described in the literature to date.

The process for the preparation of the compounds according to the invention consists in that the derivatives of 4-X-2-oxobenzothiazolines of the formula II

R ¹ = H, halogen,

NO2, alkyl,

CFs, alkoxy,

R = alkyl, (CH _{2] n} OOCR ^z, (CH) _n OCNHN (R 2) _2, (CH _{2) n} C (OR 2) = NH.HX, _{(CH2) n} OSCSR2, (CH 2) _n O, CC -morpholino, (CH 2) _n N ⁺ (R) 3X-, (CH ₂ ) _n (COOOCR 2) = NH η = 1 to 4;

R ² = alkyl, such as fungicides, in particular 4-substltuova-

where

X is the same as in Formula I, reacted with YCHaCOOCHaCeHs, where

X stands for chlorine or bromine in organic solvents such as lower aliphatic alcohols, ketones and tetrahydrofuran at 60-80 degrees Celsius for 2-6 hours in the presence of triethylamine, triethylamine and potassium iodide, potassium hydroxide or potassium hydroxide and potassium iodide . The reaction is indicated by the following scheme:

/ · * ·.

X

X where

X and Y are as described above.

Procedures for alkylation of 4-X-2-oxobenzothiazolines

Example 1

5.8 g (0.1 mol of potassium hydroxide) were dissolved in heat in 80 ml of ethyl alcohol

15.1 g (0.1 mole of 2-hydroxybenzothiazole) 18.4 g (15 ml, 0.1 mole) of benzyl chloroacetate] were added dropwise with stirring, and the reaction mixture was heated at 90 degrees Celsius in a water bath for 6 hours. After cooling, the reaction mixture was poured onto ice, the crude product was isolated and crystallized from 80% ethanol or methanol, mp 99-102 degrees Celsius, yield 19 g (63%).

Example 2

To a solution of 15.1 g (0.1 mol) of 2-hydroxybenzothiazole in 100 ml of acetone or tetrahydrofuran was added 10.1 g (14 ml, 0.1 mol) of triethylamine and 16.6 g (0.1 mol) of potassium iodide. . While stirring, 18.4 g (15 mL, 0.1 mol) of benzyl chloroacetate were added dropwise, and then the reaction mixture was heated to 60 degrees Celsius in a water bath for 2 hours. After pouring into crushed ice, the crude product was isolated and crystallized from 80% ethanol or methanol. T. t. 100-102 degrees Celsius, yield 25.4 g (85%).

Example 3

5.6 g (0.1 mole) of potassium hydroxide and 15.1 g (0.1 mole) of potassium iodide were dissolved in heat in 80 ml of ethyl alcohol or methanol, and 18.4 g (15 ml) were added in portions with stirring. , 0.1 mol) of chloroacetic acid benzyl ester or 22.9 g (0.1 mol) of bromoacetic acid benzyl ester. The mixture was heated at reflux for 2 hours and poured onto crushed ice after cooling. Crystallization from 80% ethyl alcohol. Yield 23.4 g (78.4%, mp 100-102 ° C). C 16 H 13 NO 3 S M. h. 299.35

Analysis:% C

Calculated: 64.19

Found: 64.08 ¹ H NMR of 3-benzyloxycarbonylmethyl-2-oxobenzothiazoline:

4.85 (NCH 2, s, 2H),

5.14 (COOCH 2, s, 2H),

7-7.7 (ar, m, 9H).

solubility:

methanol, ethyl alcohol, acetone, benzene, dimethylformamide, dimethylsulfoxide.

Example 4

18.5 g (0.1 mol) of 2-hydroxy-4-chlorobenzothiazole were dissolved in 150 ml of acetone or tetrahydrofuran while warming,

10.1 g (14 ml, 0.1 mol) of triethylamine and 18.4 g (15 ml, 0.1 mol) of benzyl chloroacetate were added dropwise with vigorous stirring. The reaction mixture was then heated to reflux for 4 hours in a water bath. After pouring on ice, the product was crystallized from 80% ethyl alcohol or methanol. T. t. 140-142 ° C, yield 21.6 g (65%).

% Η% N% S

4.37 4.67 10.71

4.33 4 62 10.85

Example 5

To a solution of 18.5 g (0.1 mol) of 2-hydroxy-4-chlorobenzothiazole in 150 ml of acetone was added 10.1 g (14 ml, 0.1 mol) of triethylamine and

16.6 g (0.1 mol) of potassium iodide. 18.4 g (15 ml, 0.1 mole) of chloroacetic acid benzyl ester were added dropwise with vigorous stirring, and the reaction mixture was heated to 60-70 ° C for 4 hours. After cooling, the reaction mixture was poured onto ice and the isolated product crystallized from 80% ethanol. T. t. 139-142 ° C, yield 30 g (90%).

Example 6

5.6 g (0.1 mol) of potassium hydroxide and 18.5 g (0.1 mol) of 2-hydroxy-4-chlorobenzothiazole were dissolved in 100 ml of ethyl alcohol or methanol. 16.6 g (0.1 mol) of potassium iodide were then added and 18.4 g (15 ml, 0.1 mol) of benzyl chloroacetate benzyl ester or 22.9 g (0.1 mol) of benzyl ester were added dropwise with vigorous stirring. bromoacetic acid. The mixture was heated 4 hours at 80 C ^and after pouring onto ice, the product crystallized from 80% ethyl alcohol Joint. T. t. 139-142 ° C, yield 26.8 g (80.6%).

C16H12CINO3S M. h. 333.79

analysis: % C % H % N % with % Cl calculated: 57.57 3.62 4.19 9.60 10.62 found: 57.68 3.64 4.28 9.46 10.59

^{Χ Η} NMR of 4-chloro-3-benzyloxycarbonylmethyl-2-oxo-benzotiazolínu:

5.10 (NCH 2, s, 2H),

5.16 (COOCH 2, s, 2H),

7-7.7 (ar, m, 8H).

m-multiplet, s-singlet solubility ::

methyl alcohol, ethyl alcohol benzene, acetone dimethylsulfoxide, dimethylformamide.

¹ H NMR spectra were recorded on a TESLA BS 487 A 80 MH _Z instrument in deuterated dimethylsulfoxide with hexamethyldisiloxane standard.

Chemical growth and partitioning regulators with high biological activity and selectivity today have a leading role in the directed regulation of vital plant processes. They are used to increase the yield of plants (Šebánek, J .: VŠZ, Brno, 2, 322, 1965; Nátr, L., Kousalová, I .: Study information - basic and auxiliary sciences in agriculture. ÚVTI, Praha, 1973; Kuttna , J .: Growth Regulators and its Use in Agriculture and Horticulture, SZN, Praha, 1977) and in the study of differentiation and dedifferentiation of plant cells, tissues and organs in "In vivo" and "In vitro" conditions (Thimann, ,V. : The Auxins, In, Wilkins, B .: The Physiology of Plant Growth and Development, Mc Graw-Hill, London, p. 3, 1969; Gamburg, KZ: Biochimija auxina i jego Acta na kletky rastenij Nauka, 272 (1976) Reinert, J. Bajaj, YPS: Plant Cell, Tissue and Organ Culture, Springer-Verlag, Berlin, Heidelberg, New York, p-803, 1977). The mechanism of action of phytohormonal substances on integrating life processes - growth and transformation of plant organism is mentioned by several authors. Kefeli V. I .: Growth of Growth. Moscow. Kolos. 1973, p-120, assumes that a number of phytohormones are involved in the control of each physiological process. Auxins act to shift the ionic equilibrium of forces, activate m-RNA synthesis, stimulate enzyme synthesis, modify cell wall composition, softening and cell expansion due to cell wall decalcification. Gibberellins act on the synthesis of new RNAs in the nucleus, synthesis of proteins, enzymes and activate cell growth. Cytokinins directly affect m-RNA synthesis and enzyme synthesis, activate cell growth and activate t-RNA.

Current knowledge of the mechanism of action of phytohormones suggests that their main effect is in the field of the genetic code, where they act as derepressors and repressors of enzyme synthesis, thereby interfering with the transfer of genetic information in the transcription and translational processes according to the scheme:

DNA -, - - RNA--.

whites on —-> ^r '&

(I) or act eipigenetics, by modifying the product of proteosynthesis according to the scheme:

DNA ---> RNA ----> Stable and -----> raSi>. f / O (Gamburg Z. K., Biochimija auxina i jego dejstvije na kletki rastenij. Izd. Nauka, 1976, p-271). Another idea (Butenko RG, Experimental morphogenesis and differenciacija in culture kletok rastenij; In: 35 alegod. Timirjazev. Read. Nauka, pp 3.1975) assumes that phytohormonal agents activate the proliferation of differentiated cells in the transformation process by the following mechanisms: permeability of cell structures, their properties change. This is followed by activation of the synthesis of cytoplasmic proteins that are transported to the nucleus. They specifically bind with histones, repress the loci responsible for the synthesis of r-RNA and t-RNA. DNA replication begins only when a certain threshold level of RNA in the cell is reached. This is sufficient for the cell to begin to divide.

with

The essence of the directed inferentially differentiated transformation of cells and tissues of the plant organism "In vivo" and "In vitro" consists in treating the plant material with 3-benzyloxycarbonylmethyl-2-oxobenzothiazoline or 4-chloro-3-benzyloxycarbonylmethyl-2-oxobenzothiazoline. Evidently thiazoline or 4-chloro-3-benzyloxycarbone remains after treatment with the substance in a concentration of 10 ^-4 to 10 ^-3 mol. Prolonged growth of the stem and primary root is inhibited visibly. The meshes coarse in the prolongation zone. Morphoses appear on the apexes of the stem and root on which I differentiate! side roots. As a result of the faster growth of the inner part of the tissues, the apex surface layers, the differentiation and prolongation zones and the so-called. "Root truncation", the separation of superficial cortical layers from the inner stomach part of the cell which they intensively proliferate. In the exposed differentiation zone, due to inhibition of primary root growth, the side roots grow densely side by side and form sawtooth or fan-like morphous formations with a typical terematical form of organization.

Amorphous calluses are produced by dedifferentiation under "In vitro" conditions. At the stage of the primocalus, the tissue is disintegrating into cells. After inoculation with fresh medium, analogous to that of the ear during the ear differentiation, it retains typical unorganized growth. After application of the subject substances to callus cells, the fresh biomass weight during persistent dedifferentiation increases, depending on the concentration and exposure time of the substance with a maximum efficiency at a concentration of 10 ⁵ mol. dm ^-3 . The compounds applied according to the invention cause morphogenic processes and morphological effects known by the efficacy of hormonal effectors under the conditions of &quot; in vivo &quot; and &quot; in vitro &quot;.

The degree of efficacy of the subject compounds depends on the concentration used. Said biological effects highlight the importance of 3-benzyloxycarbonylmethyl-2-oxobenzothiazoline and 4-chloro-3-benzyloxycarbonylmethyl-2-oxobenzothiazoline as potential inducers of directed dedifferentiation transformation of organism, tissues and plant cells for biotechnological manipulation.

Example 7

In the first series of experiments under the conditions of "In vitro", the plant germ (Vicia sativa L. var. Fatiina; Vicia faba L. var. Inovec; Pisum sativum L. var. Emerald) was used to induce dedifferentiation. After inhibition (6-16 hrs) in distilled water, the seeds germinated in expanded nacre, in the dark in a thermostate for 72 hrs. After the perlite wash, the germs were dried with cotton wool, selected and 25-30 mm + 1 ^mm long germs were used for further tests. The germ files (5 pieces) were planted horizontally on moistened filter paper in medium sized petri dishes. 17 cm. The filter paper was saturated with the active ingredients in concentrations of 10 ² , 10 ³ , 10 ⁴ , 10 ⁵ , 10 ^-6 , 10 _{-10 and 10} -9 _mol . . dm ' ³ . A controlled series of germs was incubated on filter paper which was saturated with distilled water. Induction of dedifferentiation was performed in a thermostate in the dark at 25 ° C + 1 ° C for 72 to 96 hours. The effect of dedifferentiation transformation was determined visually using a microscopic magnifying glass. The maximum dedifferentiation efficacy is due to the test substance on the plant germs after treatment with the substances in concentrations of 10 ^-4 and 10 ^-3 mol. dm- ³ .

Example 8

In a second series of experiments under "in vitro" conditions, calogenesis was induced on apical segments of primary roots of the lid (Vicia sativa L. var. Solarka). The selected seeds (approximately the same size and color of osmosis) were sterilized with 5% chloramine solution for 1 hour and rinsed several times with sterile distilled water. The sterile seeds were germinated in Petri dishes to 0.8% in agar medium Sterile germs 25-30 millimeters + 1 mm long were used to induce calogenesis, and apical primary root segments of 10-15 mm length were decapitated and planted into Petri dishes in horizontal position at 0.6% modified solid agar medium according to Murashige. In the experimental variations, the medium contained the active ingredient in a concentration of 10 ^-2 to 10 ^-9 mol dm ^-3 .

2,4-D at a concentration of 10 ^-5 mol. dm ^-3 . Calogenesis was induced at 25 ° C, in the dark for 14 days. Preparation of the test material as well as cultivation were performed under aseptic conditions. The maximum dedifferentiation efficiency was found at a concentration of 5.10 ⁴ mol. dm ^-3 .

Example 9

In the third series of experiments, the effect of the substances on the formation of the fresh weight of the callus tissue of Haplopappus gracilis was determined. Callus from culture found in passage 53 grown on solid agar medium according to Murashige-Skooga was used as the inoculum. The initial inoculum weight ranged from ^1.1-2.3 g ^-1 . In the test series, the inoculum was seeded in 100 ml Erlenmeyer flasks on solid agar medium according to Murashige-Skooga with an active substance content of 10 ⁷ , 10 ^-6 , 10 ^-5 , 10 ^-3 mol. dm ^-3 in individual variants. The control series contained, as a phytomormonal effector, 2,4-D at a concentration of 10 ^-5 mol. dm ' ³ . The cultivation was carried out in a thermostate in the dark at 25 ° C. Samples were taken on 7, 14, and 21.

232502 day of cultivation, in 10 reps. Fresh weight production was evaluated gravimetrically. (See graph). The maximum increase in fresh weight was achieved after treatment with the active compound at a concentration of ¹ mol of ⁵ mol. . dm ^-3 . 4-C1-3-benzyloxycarbonylmethyl-2-oxobenzothiazoline showed the same efficacy.

The investigated substances cause analogous morphogenic processes and morphological effects as phytohormones under "In vivo" and "In vitro" conditions. The fresh weight of biomass after drug challenge to callus cells increases as a function of concentration and time of drug challenge with a maximum efficiency at a concentration of 10 ^-5 moles. dm ' ³ .

Chart legend:

Fresh callus weight gain due to different concentrations of 3-benzyloxycarbonylmethyl-2-oxobenzothiazoline.

control solution of 10 ^-3 M solution of ICs .l ^{-1 3} mol. I ^-1 solution of IO- ⁶ mol. 1 ^-1 solution of 10 ^-7 mol. I ¹

Claims (2)

SUBJECT CLAIMS 1. 4-X-3-Be, nzyloxycarbonylmethyl-2-oxobenzothiazolines of the general formula I: X is hydrogen or chlorine. 2. A method according to claim 1, wherein the 4-X-2-oxobenzothiazoline derivatives of the formula. II INVENTION * 00 where X is the same as in formula I, reacted with a compound of formula (II) YCH2COOCH2C6H5 where Y represents chlorine or bromine, in an environment of organic solvents such as aliphatic alcohols, ketones, and tetrahydrofuran at 60-80 ° C, for 2-6 hours in the presence of triethylamine or trlethylamine and potassium iodide or potassium hydroxide or potassium hydroxide and iodide potassium.