CS270539B1 - 3-Allyl-6-nitro-2-benzothiazolinone and its preparation - Google Patents

Abstract

The previously unknown 3-allyl-6-nitro-2-benzothiazolinone of formula I is obtained by the reaction of 6-nitro-2-benzothiazolinone in the presence of an equivalent amount of potassium hydroxide and allyl bromide in a mixture of water and ethanol under reflux. The compound, depending on the concentration, inhibits and re. stimulates the growth and synthesis of chlorophyll in green algae of monocotyledonous and dicotyledonous plants and callus tissue cultures, in which it supports the formation of plant organs.

Description

The present invention provides 3-allyl-6-nitro-2-benzothlazolnon and a process for its preparation.

To date, 3-aubatitated 6-nitro-2-benzothiazolnonone derivatives have been known as plant growth regulators (D'Amico 3.3., U.S. Pat. 4,282,029 (1981). Said compounds contain a substituent with a nitrogen heterocycle in position 3. 3-Substituted 6-nitro-2-benzothiazolinone derivatives are also known as herbicides. substances contain an amido group in the substituent in position 3 [D'Amico 3.3., U.S. Pat. 4,185,990 (1980). Of the simpler derivatives, 3-alkyl-6-nitro-2-benzothiazolinones with saturated alkyls are known.

We have now glorified that hitherto unknown compound of the formula

depending on the concentration inhibits the growth and synthesis of chlorophyll of green algae, monocotyledonous and dicotyledonous plants of callus tissue cultures, 3als that the compound of the formula IIS in a wide concentration range (10 to 10 mol.dm) tissue cultures and at the same time supports the formation of plant organs in kaluoova plotivovoj culture analogous to kinetin.

At the same time, the golden process for the preparation of the title compound was reacted with 6-nitro-2-bonzothiazolinone [Sidóová, E., author no. 265,776 / in the presence of a quantitative amount of potassium hydroxide and a weight-equivalent quantity or up to a 20% excess of allyl bromide in an aqueous-alcoholic medium at reflux for 15 minutes to 3 hours.

A 1: 2 mixture of water and ethanol is preferred.

The following examples further illustrate, but in no way limit, the preparation and properties of the compound of the invention.

Example 1

Preparation of 3-allyl-6-nitro-2-benzothiazolnon

6-Nitro-2-benzothiazolnon (5.9 g, 0.030 mol) was dissolved in a solution of potassium hydroxide (2 g, 0.030 mol) in a mixture of water (10 cm ³ ) and ethanol (20 cm ³ ). Allyl bromide (4.0 g, 0.033 mol) was added to the solution and the reaction mixture was refluxed for 2 hours. The crude product, mp 137-14 ° C, was obtained in an amount of 5.7 g (80.5%). The material was recrystallized from benzene using activated carbon, the pure product was extruded from the benzene solution with cyclohexane. Highly pure 3-allyl-6-nitro-2-benzothiazolnon, m.p. 147.5-148.5 ° C, was obtained in an amount of 4.1 g (57.9%). Mh: 236.25 For C ₁₀ H ₈ N ₂ 0 ₃ S calculated: C 50.84%, H 3.41%, N 11.86%, S 13.57% fused: C 60.99%, H 3 , 32%, N 12.00%, S 13.83%

Example 2 '

Effect of a compound of the invention on the growth and content of synthesized chlorophyll of the alga Chlorella vulgaris

Algae Chlorella vulgarie ea were pasted by stionionation by culturing at 25 2 1 ° C in a light regime: 16 hours light, 8 hours dark in a cultivating pad according to / shark, I., A.R. Algologist. Lab., Třeboň 1967: 71-140, Třeboň (1968) /. The test substance was added to the culture medium in dimethylformamide solution, and the same volume of solvent was added to the control samples. Samples were inoculated with the exact volume of homogeneous

CS 270 539 91 algal cell suspensions. After culturing for 7 days, the suspension! cells were measured by absorbance at 660 nm (proportional to algal cell weight) and chlorophyll content after extraction! dimethylformamide.

The compound of the invention at higher concentrations (1.10 ⁴ M) significantly inhibited the growth and synthesis of chlorophyll algae Chlorella vulgaris (44.0 + 2.3% compared to 100% control) at concentrations of 1.10 - 1.10 M for a slight stimulating.

Influence of the compounds according to the invention on the raet and chlorophyll content of the algae Chlorella vulgaris.

Concentration% Control β ₃ Absorbance at chlorophyll content mol / dm 660 nm

1.10 ⁴ 60.1 ± 0.6 44.0 ± 2.3 1.10 “ ⁵ 105.2 2 12.3 107.1 ± 20.5 1.10 “ ⁷ 108.2 ± 5.2 104.9 + 6.0 1.10 ⁹ 99.9 + 9.1 * mm · ’ 105.2 ± 8.5 1.10 ¹¹ 102.2 ± 2.5 103.9 + 3.3 1.10 “ ³ 99.4 + 7.1 104.4 ± 11.3

»Deviation from the average

Example 3

Test of the phytotoxicity of the compound according to the invention and its effect on the growth of fresh matter as well as the content of synthesized chlorophyll in the aboveground part of maize (Zea mays L.).

The phytotoxicity of the compound according to the invention was evaluated by measuring the prolongation growth of the root and stem (Murin, A., Koleková, A., Váchová, M. and Regula, š., Biologie / Bratislava) 35, 937 (I960); Sekerka, V. and Sutoris, V., Agriculture 31, 872 (1985). Maize / Zea mays L., variety TO 560 A /, year of harvest 1986, supplied by the Regional Seed Company of Trnava, was used as the material.

The corn seeds swelled in the solutions at the concentration given in the table for Example 3 for 24 hours. They were then placed on a moistened filter paper, wrapped in perforated polyethylene bags and allowed to germinate in a mandrel at 25 ° C + 1 ° C (ČSN 46 0610). After 72 hours of growth, the length of the roots and the aboveground part were measured. Distilled water was used as a control. The results were statistically evaluated by calculating the arithmetic mean, standard deviation and confidence interval / ISq _Q5 / for 95% probability / Reisenauer, R., Methods of Mathematical Statistics and Leh application in technology, SNTL Prague (1970), p. 207 /.

The highest concentration used (1.10 ⁴ ml.dm ³ ) had an inhibitory effect on the root length, the average length of which was only 24.75 ± 1.50 mm, which is 52.67% of the control. Stem length was only slightly below the control value (20.30. + 1.24 mm) compared to 22.03 2 θ, 31 mm for the control, which represents 92.15 5.63% of the control.

All other concentrations of the compound according to the invention used had a stimulatory effect, to a greater extent on the aboveground part than on the roots. The highest stimulation was recorded at a concentration of 110% mol.dm 3, where the measured root length (75.29 + 0.79 mm) represents 160.23 + 1.68% of the control. The width of the 48.92 + 2.70 mm stem compared to 22.03 2 0.31 mm of the control represents 222.06% of the control.

When testing the increase in fresh mass as well as the content of synthesized chlorophyll

CS 270 539 B1 in the above-ground part of the corn and the corn seeds were pre-germinated in the dark in rolls of rotating filter paper moistened with distilled water for 3 days. The ea was then placed in hydroponics containing the nutrient solution according to Hoogland / Hoogland, o. R., Snyder, W. C., Hoogland’s inorganic salt solution and A - Z supplement in Hewitt, E. □. / Ed / Sand and water culture methods used in the study of plant nutrltion-Nutrition Commonwealth agric. Bur ·, Farnham Hoyal, etr. 189 (1968) / reap. The effect of the compound according to the invention was tested at concentrations of 10 and 10 mol.dm, the concentration of dimethyl sulfoxide in the test and control samples was 0.005%. After 6 days of cultivation, the weight of the green part of the seedlings and the chlorophyll content were determined.

At the concentrations of the compound according to the invention tested, ea- showed a stimulating effect on both the growth of the fresh mass and on the circulation of the synrophyl synthesized in the aboveground part of maize.

Table for example 3

Effect of a compound of the invention on the prolonged growth of Zea mays L. roots and stems after 72 h. thorn incubation.

Mol.dm concentration “ ³ The root of the root Oížka stalks mm + I.S * '0.05 % + Is _c >, 05 mm + I. ,WITH, '0.05 % + I.S ⁵ 'C 1.05 0 / control / 46.99, + 3.49 100.00 ♦ 7.43 22.03 4 «Μ 0.31 100.00 4 • M 1.41 1. 10 “ ¹³ 75.29 ♦ * «Μ 0.79 160.23 1.68 48.92 4 M 2.70 222.06 4 12.26 1. 10 “ ⁹ 69.91 + 0.63 148.78 4 M 1.34 48.51 4 • M 2.98 220.20 _ + 13.53 1. 10 “ ⁷ 60.15 + 1.69 128.01 4 M 3.38 48.51 4 ee 2.39 211.12 4 * 10.85 1. 10 “® 58.83 + 2.34 125.20 4 SMI 4.98 42.79 4 • m 3.08 194.24 4 13.98 1. 10 “ ⁴ 24.75 + am 1.50 52.67 4 3.19 20.30 4 • mt 1.24 92.15 4 5.63

ISq ₀₅ interval epolahllvoeti for 95% probability.

Effect of a compound according to the invention on the growth of fresh matter and the chlorophyll content in the aboveground part of maize.

Concentration • j -3 mol .dm

About / control /. 10 ⁶ . 10 “ ⁸

Percentage increase compared to control

- ^{I, S,} 0,05 weight of green platoon chlorophyll content of seedlings

100.0 ♦ 7.95 100.03.72

127.6 * 8.82 117.1 20.66

119.3 ^ 6.32 121.5. + 3.17

ISq ₀₆ «confidence interval over 95% probability.

CS 270 539 Bl

Example 4

Influence of the compound according to the invention on the growth and chlorophyll content of the callus tissue culture of carrots (Caucus carota L.) using a complete nutrient medium

The material used was a carrot tissue culture (Caucus carota L.) isolated at the Chemical Institute of Comenius University, which was cultured on a liquid nutrient medium (Muraschige, T. and Skoog, F., Physiol. Plant. 15, 473 (1962) / using a paper knife / Erdelský, K., Acta Tao. Rer. Nature. Univ. Comenianae, Physiologia platarun 3, 1 (1971)] and by the addition of 0.1 mg dm of kinatin, 1.0 mg dm of 2,4-dichlorophenoxyacetic acid and 2% sucrose. To this nutrient medium was added a compound of the invention, the final concentration of which is given in the table to Example 4. Cultivation was carried out under a light of 7,000 lux, photoperiod 16 hours, 26 ° C £ 1 ° C, subculturing time 21 days in three passages. . The results were statistically evaluated.

The compound of the invention at a concentration of 1.1 ^-4 mol.dm ³ showed 60% growth inhibition with a concomitant increase in dry matter circulation. Total chlorophyll obeah decreased by 27%

Table for example 4

Influence of the compound according to the invention on the properties of the callus] tissue culture of Caucus careta L. after 21 days of subcultivation% of control + IS _O Concentration *

mol.dm IR +: «* ^ * ^S * 0.05 chl / dr. 1 ^{I, S,} 0.05 % t X I.S uš * 0.05 IR chl / dr. 0 / control / 1.43 + 0.04 0.2372 + 0.01 4.14 £ 0.07 100; 00 Jh 0.03 100.00. *. 4.22 1 . 10 ¹³ 3.60 + 0.04 0.8751 + 0.02 3.33 + 0.10 252.04 + 0.03 368.02 +, 8.43 1 . 10 “ ⁷ 2.60 + «T 0.02 0.7021 + 0.01 3.59 + 0.11 182.04 + 0.01 295.42 £ 4.22 1 . 10 ⁴ 0.86 + 0.04 0.1731 0.01 6.57 1 ° · ⁰⁹ 60.14 + 0.03 72.98. + 4.22

IR - growth index, chl / dr. - mg chlorophyll / 100 g dry matter; ^z * ^s * o 05 ^interv al reliability for 95% probability.

on average, while in the first passage it still reached the value of control and in the following gradually decreased.

The compound according to the invention at concentrations of 1.10 and 1.10 -mol.dm highly stimulated the growth index as well as the chlorophyll content, which reached up to three times the amount. Higher stimulation of chlorophyll content corresponds to. Example 3, where a higher stimulation of the aboveground rest of maize was found.

Example 5

The effect of the compound according to the invention on the growth and chlorophyll content of the callus tissue culture of carrots (Caucus carota L.) when used in combination with kinetin

Carrot callus tissue culture (Caucus carota Li) was used as material - culture conditions according to Example 4.

Standard nutrient medium (used as a control) to maintain disorganized growth of callus tissue cells contained a combination of kinetin (4.65). lO ⁷ mol. dm) and 2,4-dichlorophenoxyacetic acid (4.62.10 mol.dm). The compound of the invention (final concentration in the nutrient medium is given in the table to Example 5).

CS 270 S39 B1 5 was used in combination with kinetin to exclude 2,4-dichlorophenoxyacetic acid from the nutrient medium. '

In all cases, an increased growth index as well as chlorophyll content was observed, with higher values for the calf being higher at lower concentrations.

Example 6.

Influence of a compound according to the invention on the growth and circulation of chlorophyll callus tissue culture of carrots (Daucus carets L.) in the case of its use as the sole growth regulator in the nutrient medium

Carrot callus tissue culture (Daucue carota L.) was used as the material - culture conditions according to Example 4.

The compound of the invention, added to the nutrient medium without other pastes in varying concentrations (the resulting concentration in the nutrient medium is given in the table in Example 6) caused an increase in growth index compared to the control of 10 to 30% at all concentrations, while the chlorophyll content was stimulated only using very low concentrations (0.0) of the compound of the invention.

Example 7

The stimulating effect of the compound according to the invention on the formation of plant organs in the callus tissue culture of carrots (Caucus carota L.) when used as the sole growth regulator resp. in combination with kinetin

The effect of the compound of the invention on the formation of plant organs in the callus tissue culture of Caucus carota L. when used as the sole growth regulator resp. in combination with kinetin, after 21 days of subculturing.

Experimental variants Organ formation /% +, ^{I, s,} o ₍ Q5 / K + 2,4 - D 0 2.4 - 0 0 K 100.00. + 1.23 A 78.75 ♦ 1.34 8 102.64 + 1.47 C 87.75 + 1.24 D 75.43 + ^r M 1.20 K + A 81.25 2 1.87 K + B 106.24 + 1.46 K + c 108.17. + 1.54 K + D 112.49 2 1.57 -7 —3 K - kinetin in the amount of 4,65.10 mol.dm, 2,4-0 - 2,4-dichlorophenoxyacetic acid v

amount of 4.52.1O “ ⁶ mol.dm“ ³ , A, B, C, D - compound according to the invention in an amount with A - 1.10® mol.dm “ ³ , B - 1.10 ⁷ mol.dm“ ³ , C - l.lO ¹³ mol.dm “ ³ , D - l.lo ^1S mol.dm ³ , IS _n confidence interval for 95% probability.

v f UO

Carrot callus tissue culture (Daucus carota L.) was used as the material under the culture conditions of Example 4.

CS 270 539 Bl

Table for example 5

Influence of the compound according to the invention on the properties of the callus culture of Daucus carota L. when used in combination with kinetin, after 21 days of subculturing

Experimental variants IR 2 ^ * ³ * 0.05 Chl / dry. 2 % dry matter % control 2 IS _{O O5} 1 0.05 + I.S. «» '0.05 IR chl / dr. control 2.04 1 0.05 0.8008 2 0.03 2.68 + 0.10 100.00 +, 2.45 100.00 + 3.75 A 2.26 2 0.03 0.9141 2 0.03 2.94 2 0.06 111.45 ± 1.47 114.15 + 3.75 B 2.39 ♦ 0.04 0.9826 + 0.02 2.53 i 0.04 117.58 2 1.96 122.71 2 4.50 C 2.76 + 0.07 1.0041 1 0.02 2.69 + 0.02 135.28 ♦ 3.43 125.39 2 3.75 0 '2.80 + 0.04 M 1.0185 + 0.03 2.74 + 0.04 137.47 ± 1.96 127.18 2 3.75

IR - growth index, chl / dr; - mg chlorophyll / 100 g dry matter; Control - kinetin in the amount of 4.65. 10 mol.dm + 2,4-dichlorophenoxyacetic acid 4.52. 10 mol.dm; all variants of the experiment / A, B, C, 0 / contain kinetin in the same amount as the control, supplemented with the S-2 '-7 -3 -13 compound according to the invention in the amount: A - 1.10 mol.dm, B - 1.10 mol .dm, C - 1.10

«2 2 mol.dm, 0 - 1.10 m.ol.dm, IS _{Q og} confidence interval for 95% probability.

Table for example 6

Influence of the compound according to the invention on the properties of the callus tissue culture of Daucus carota L. when used as the sole growth regulator, after 21 days of subculturing

Experimental variants IR 2 ^I ' ^S ' 0.05 chl / dr. 2 ... t e % dry matter % control 2 IS _{0 (05} 2 i.s. '0.05 0.05 IR chl / dr. control 2.04 2 0.04 0.8008 2 0.03 2.68 * • * «* 0.09 100.00 2 1.96 100.00 2 3.75 2.4-0 2.08 2 0.05 0.6567 2 0.03 2.76 2 0.07 101.96 2 2.45 82.08 2 3.74 kinetin 2.38 2 0.04 0.9099 2 0.02 2.95 2 0.06 116.67 2 1.96 111.89 2 2.50 A 2.25 + 0.07 «Μ * 0.7159 + 0.01 • M * 3.27 + 0,0,7 110.29 + 3.43 89.95 + 1.25 B 2.62 + 0.06 «Μ * 0.7650 ♦ 0.02 2.98 2 0.06 128.43 2 2.94 95.53 2 2.50 C 2.68 + 0.04 Mt * 0.9993. 2 0.03 2.68 + 0.04 131.37 2 1.96 124.79 2 3.75 'D 2.37 2 0.06 0.9365 + 0.02 * 2.59 2 0.01 116.20 + 2.94 116.95 2 2.50

IR - growth index, chl / dr. - mg chlorophyll / 100 g dry matter; Control - kinetin in the amount of 4.65.1O'mol.dm + 2,4-dichlorophenoxyacetic acid in the amount of 4.52.10 mol.dm, t —6 * 2 ’

2,4-O-dichlorophenoxyacetic acid in an amount of 4.52.10 mol.dm, kinetin-kinotrin in an amount of 4.65.10 mol.dm; Variants of experiments A, B, C, D contained the compound according to

-12 of the lesion with the final concentration: A - 1.10 mol.dm, B - 1.10 mol.dm, C - 1.10 mol.dm ³ , 0 - 1.10 ^1S mol.dm ³ .

!.WITH. _{O 05} confidence interval for 95% probability.

CS 270 639 Bl

The compound according to the invention, added to the nutrient medium in a wide concentration range alone or together with kinetin, caused morphological differences compared to the control, in the first pass the calli grew without visible change, with a typical lumpy structure. In the second passage ea, on the surface of the calli, deep green dots began to appear, from which the foundations of roots and stems gradually developed. In the third stage, organogenesis reached the stage of well-developed young carrot plants. Their amount was evaluated in comparison with a nutrient medium containing separate kinetin, where it represented 100% of the formation of young plants. Raatlin organogenesis did not occur on the nutrient medium with the addition of 2,4-dichlorophenoxyacetic acid alone or on the standard nutrient medium (combination of 2,4-dichlorophenoxyacetic acid and kinetin). The calluses have a typical lumpy structure.

Importantly, the compound of the invention at higher concentrations inhibits the raat and chlorophyll synthesis of green algae, monocotyledonous and dicotyledonous plants and callus tissue cultures. Of particular interest is the fact that the compound of the invention in low _- e. «IR 'concentrations in a wide range (10 mol.dm to 10 mol.dm) stimulate the growth and synthesis of chlorophyll plants and callus tissue cultures and at the same time promote the formation of plant organs in callus tissue culture analogously to kinetin, so that it cannot be completely replaced.

The compound according to the invention can be used as an eco-stimulator of plant growth and callus tissue cultures here or in combination with other substances, and can be used pre-emergently or post-emergently for plants. The salt compound of the invention can be used for merithematic micropropagation of plants as an equivalent substitute for kinetin. In addition, the compound of the present invention can be used as an active ingredient of anti-allergic preparations or as an intermediate for 3alSie syntheses.

Claims (2)

The compound of the invention, added to the nutrient bladder in the Siroke Concentration Scale, or together with Kinetin, induced morphological differences over control. In the first passage, calluses grew without visible change, with a typical lumpy structure. In the second passage and on the surface of the kalua, they began to discover green points, from which they developed the foundations of roots and stems. In the third passage, the organogenesis has achieved a good level of developed young carrot reptiles. Their amount was compared with a nutrient bladder containing a separate clinker, where the formation of young plants was 100%. Organogenesis did not occur on the nutrient medium with the addition of 2,4-dichlorophenoxyacetic acid alone on standard nutrient / 2,4-dichlorophenoxyacetic acid and kinetin / nutrient. Kaluey had a typical lumpy structure. Significant is the fact that the compound of the invention at higher concentrations of inhibition and synthesis of chlorophyll of green algae, monocotyledonous and dicotyledonous plants and callus tissue cultures. Of particular interest is the fact that the compound of the invention in low e · 3 · 3 concentrations over a wide range of 1010 mol.dm to 10 mol. plant organs in callus tissue culture analogously to kinatin, so that it can be replaced. The compound of the present invention can be used as a plant growth and callus growth factor stimulator alone or in combination with lipids, for pre-emergence or post-emergence plants. In the violet compound of the present invention, a premixed mlcropropagmatic plant can be used as an equivalent substitute for clavin. In addition to the aforementioned compound of the present invention, it is possible to use as an active ingredient antiallial agents or as an msdzlproduct for 3alpha synthesis. OBJECT OF THE INVENTION 1. 3-Allyl-6-nitro-2-benzothiazolinone of Formula C = O ' 2. A process for the preparation of a compound according to claim 1, characterized in that the 6-nitro-benzothiazolinone is reacted in the presence of an equivalent amount of potassium hydroxide and equilibrium or up to 20% by weight. excess amount of allyl bromide in a mixture of water and ethanol at reflux temperature.