JP2023164356A - Inhibitor for activity of auxin and anthranilic acid - Google Patents

Abstract

To provide a novel inhibitor for the activity of auxin and anthranilic acid, and a plant growth regulator.SOLUTION: Provided is an inhibitor for the activity of auxin and anthranilic acid that contains a compound represented by the following formula 1 or a salt thereof as an active ingredient (where, A is a single bond via -CH(=O)-NH- or -CH=CH- structure, R1 is any of a hydrogen atom, a halogen atom, or an alkyl group, R2 is any of a hydrogen atom, a halogen atom, or an alkyl group having 1 to 6 carbon atoms, R3 is a hydrogen atom, an alkyl group or a halogen atom, R4 is OH or OCH3, and R5 is any of an amino group, a halogen atom, a hydroxyl group, or a hydrogen atom. R1 and R2 may form a closed benzene ring).SELECTED DRAWING: Figure 1

Description

The present invention relates to an agent that inhibits both auxin and anthranilic acid activity, and a plant growth regulator.

In the agricultural field, controlling plant growth is an important technology for improving productivity. Currently, various types of plant growth regulators are in practical use for the purpose of regulating plant growth, and plant growth regulators contribute to improving crop yields and product quality.
Among them, auxin is one of the well-known substances as a hormone that controls plant growth.
Auxins are a group of plant hormones that primarily have the effect of promoting plant growth (elongation), and are synthesized at the tips of plant stems and exert their effects while moving toward the base of the stem. Its main effects are known to be promoting stem elongation, suppressing the growth of lateral buds, elongating leaves upward, promoting fruit enlargement, promoting aging, and promoting rooting, but it is also involved in cell division itself. Therefore, it is thought that it has various indirect effects in addition to these. Indole-3-acetic acid (IAA) and phenylacetic acid (PAA) are the most abundant naturally occurring auxins, and indole-3-butyric acid (IBA) is also found in corn and other foods. Synthetic auxins include naphthaleneacetic acid, naphthoxyacetic acid, 2,4-dichlorophenoxyacetic acid (2,4-D), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), 2methyl,4chlorophenoxy Examples include acetic acid (MCP), picloram, and dicamba.
These auxins have in common that they have an aromatic ring and a carboxyl group, but when classified structurally, those with a carboxyl group attached directly to the aromatic ring (picloram, dicamba, etc.) and those with an alkylene group between the aromatic ring and the carboxyl group. groups (indole-3-acetic acid, indole-3-butyric acid, naphthaleneacetic acid, phenylacetic acid, etc.) and those in which an alkyleneoxy group is interposed between the aromatic ring and the carboxyl group (2,4 -dichlorophenoxyacetic acid, 2,4,5-trichlorophenoxyacetic acid, 2-methyl-4-chlorophenoxyacetic acid (MCP), etc.). If the action of auxin is inhibited or the synthesis of auxin is inhibited, plant elongation will be suppressed, causing problems such as dwarfism.On the other hand, if this action can be artificially controlled using plant growth regulators, , the potential for agricultural use is considered to be extremely high.

In addition, research on auxin receptors, which had been unknown for more than 70 years since the discovery of auxin, has progressed, and in 1998 it was discovered that the TIR1 (transport inhibitor response 1) protein is an auxin receptor in Arabidopsis. Non-patent document 1), followed by the homolog of this protein AFB (auxin sign
(aling F boxproteins) 1 to 5 have also been discovered, and the mechanism of action of auxin is being clarified. Among these, TIR1, AFB1, AFB2, and AFB3 have been found to be the main receptors for auxin, which has an alkylene group or alkyleneoxy group interposed between the aromatic ring and carboxyl group, such as IAA and 2,4-D. (Non-Patent Document 2), and AFB5 has been found to be a receptor for picloram (auxin in which a carboxyl group is directly bonded to an aromatic ring) (Non-Patent Document 3). Furthermore, it has become clear that homologous genes for these proteins exist in all plants, and it has become clear that the auxin action mechanism revealed in Arabidopsis is applicable to other plants as well. Furthermore, it has been revealed that tryptophan aminotransferase (TAA1) and flavin monooxygenase (YUCCA), which were previously thought to exist in different biosynthetic pathways, are actually enzymes that exist in the same pathway. The main pathway for biosynthesizing IAA from indole-3-pyruvate, an intermediate in IAA biosynthesis, has been clarified (Non-Patent Document 4).

As such knowledge regarding auxin has become clearer, research has progressed to find substances that inhibit the expression of auxin activity, and a large number of auxin activity inhibitors and plant growth chemical regulators have been proposed (Patent Documents 1 to 3). 5). Examples of auxin inhibitors include those that inhibit auxin biosynthesis, those that inhibit active transport of auxin in plants, and those that inhibit binding to auxin receptors. Among these, auxin receptor inhibitors have been developed that inhibit the normal reaction of the receptor by binding a side chain to IAA, which is a natural auxin (Non-Patent Documents 5 and 6). Therefore, these auxin inhibitors had a structure in which an alkylene group was interposed between an aromatic ring and a carboxyl group.

On the other hand, the present inventors are continuing to conduct search research for compounds having auxin action. During this process, we discovered that anthranilic acid, which was thought to be a precursor of indoleacetic acid, directly induces the development of adventitious roots and further promotes the development of the root system. They have continued to research this effect and found that derivatives having an anthranilic acid skeleton also have similar auxin activity, and have already applied for patents (Patent Documents 6 to 9). That is, anthranilic acid causes the emergence of adventitious roots in plants and develops the root system.
Because of this effect, anthranilic acid is considered to be useful as a plant growth regulator, similar to auxin.

Substances that can inhibit the activity of auxin and/or anthranilic acid in plants are also considered useful as plant growth regulators. However, until now no substance has been provided that inhibits the activities of both auxin and anthranilic acid.

<Explanation of 3D structure>
By the way, it is generally known that the activity of drugs and low-molecular physiologically active substances is related to their three-dimensional structure.
It is known that when two aromatic hydrocarbons are bonded via an amide structure, the entire carbon skeleton becomes a planar structure. In the case of an amide structure, the carbon skeleton is known to have a rigid, planar structure, and this planar structure is called an "amide plane" (see Non-Patent Document 7). It is known that this occurs because the p-orbital of the nitrogen in the amide structure overlaps with the p-orbital of the carbonyl group (see Non-Patent Document 8). Therefore, when aromatic hydrocarbons are bonded via an amide bond, their p orbitals also overlap, so that the entire carbon skeleton is arranged on the same plane. Also, when a vinylene group is substituted for an amide bond, the p-orbital of the double bond in the vinylene group overlaps with the p-orbital of the aromatic hydrocarbon, so the entire carbon skeleton is similarly arranged on the same plane. Ru.
<Explanation of relationship with activity>
For this reason, in the case of compounds whose carbon skeletons require a planar structure to exhibit biological activity, it is possible to change the partial structure while maintaining the planar structure of the molecule without changing the overall biological activity. This ability is well known as bioequivalence (bioisostere). A particularly known example in which the same activity is maintained regardless of the order of the amino group and carbonyl group in the amide bond is amide isoster (see Non-Patent Document 8).

Furthermore, it is known that the same activity is maintained even if the above-mentioned amide structure is replaced with a vinylene structure, and it is also considered to be a bioisostere (see Non-Patent Document 8). No attempt has been made to date to search for inhibitors of auxin or anthranilic acid that take into consideration the maintenance of the three-dimensional structure (planar state) of the carbon skeleton of the compound.

International Publication No. 2008/150031 JP2013-67656A Japanese Patent Application Publication No. 2014-118404 Japanese Patent Application Publication No. 2014-169246 Japanese Patent Application Publication No. 2016-169221 International Publication No. 2016/035685 Japanese Patent Application Publication No. 2017-178852 JP2018-52865A JP2018-52866A

Ruegger et al (1998) The TIR1 protein of Arabidopsis functions in auxin response and is related to human SKP2 and yeast Grr1p. Genes and development 12 198-207 Dharmasiri et al. (2005) Plant development is regulated by ad family of auxin receptor F box proteins, Developmental Cell, Vol. 9, 109-119, 2005 Walsh et al. (2006) Mutations in an auxin receptor homolog AFB5 and in SGT1b confer resistance to synthetic piclinate auxins and not to 2,4-dichlorophenoxyacetic acid or indole-3-acetic acid in Arabidopsis, Plant Physiology, Vol. 142, 542 -552 Mashiguchi et al. (2011) The main auxin biosynthesis pathway in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America, 2011, doi:10.1073/pnas.1108434108 Hayashi et al. (2008) Small-molecule agonists and antagonists of F-box protein-substrate interactions in auxin perception and signaling. PNAS, vol. 105, no.14 Hayashi et al. (2012) Rational design of an auxin antagonist of the SCF(TIR1) auxin receptor complex. ACS Chem Biol, 16;7(3):590-8 Voet and voet (2011) Three dimensional structures of proteins “Biochemistry” (4th edition), p221 Silverman and Holladay (2014). Lead Discovery and Lead Modification. In “The Organic Chemistry of Drug Design and Drug Action (3rd edition)” p71. Yang et al. (2008) Inactive methyl indole-3-acetic acid ester can be hydrolyzed and activated by several esterases belonging to the AtMES esterase family of Arabidopsis. Plant Physiology, Vol.147, pp.1034-1045

Inhibiting the activities of both auxin and anthranilic acid is thought to lead to the development of new plant growth regulators.
An object of the present invention is to provide a substance that inhibits the activity of both auxin and anthranilic acid, and an agent for regulating plant growth that includes a substance that inhibits the activity of both auxin and anthranilic acid.

That is, the present invention consists of the following configuration.
1. An auxin and anthranilic acid activity inhibitor containing a compound represented by the following formula 1 or a salt thereof as an active ingredient.

(However, A is a single bond via a -CH(=O)-NH- or -CH=CH- structure, R1 is a hydrogen atom, a halogen atom, or an alkyl group, and R2 is a hydrogen atom, A halogen atom, an alkyl group having 1 to 6 carbon atoms, R3 is a hydrogen atom, an alkyl group, or a halogen atom, R4 is OH or OCH3, and R5 is an amino group, a halogen atom, a hydroxyl group, or hydrogen (R1 and R2 may form a closed benzene ring.)
2. The auxin and anthranilic acid activity inhibitor according to 1, which contains as an active ingredient a compound, a salt or an ester thereof, in which the halogen atoms of R1, R2, and R3 are fluorine atoms, and the alkyl groups of R1, R2, and R4 are methyl groups. .
3. An auxin and anthranilic acid activity inhibitor containing as an active ingredient one or more compounds selected from the following compounds (1) to (15).
(1) 4-[(phenylamino)carbonyl]-benzoic acid (2) 2-amino-4-[(phenylamino)carbonyl]-benzoic acid (2- Amino-4-[(phenylamino)carbonyl]-benzoic acid)
(3) Methyl-2-Amino-4-[(phenylamino)carbonyl]-benzoate
(4) 2-Amino-4-[(1-naphthylamino)carbonyl]-benzoic acid
(5) 2-Amino-4-[(2,3-dimethylphenylamino)carbonyl]-benzoic acid
(6) 2-Amino-4-[(2-fluorophenylamino)carbonyl]-benzoic acid
(7) 2-Amino-4-[(3-fluorophenylamino)carbonyl]-benzoic acid
(8) 2-Amino-4-[(4-fluorophenylamino)carbonyl]-benzoic acid
(9) 4-(benzoylamino)benzoic acid
(10) 2-hydroxy-4-(benzoylamino)benzoic acid (11) 2-Chloro-4-(benzoylamino)benzoic acid (2-Chloro-4- (benzoylamino)benzoic acid)
(12) 4-(4-fluorobenzoylamino)benzoic acid
(13) 4-Stilbenecarboxylic acid
(14) 2-amino-4-(benzoylamino)benzoic acid
(15) 2-Amino-4-styrylbenzoic acid
4. A plant growth regulator comprising the auxin and anthranilic acid activity inhibitor according to any one of 1 to 3.
5. A lateral root development inhibitor comprising the auxin and anthranilic acid activity inhibitor according to any one of 1 to 3.
6. A taproot elongation promoter comprising the auxin and anthranilic acid activity inhibitor according to any one of 1 to 3.
7. A plant overgrowth inhibitor comprising the auxin and anthranilic acid activity inhibitor according to any one of 1 to 3.
8. A branching development promoter comprising the auxin and anthranilic acid activity inhibitor according to any one of 1 to 3.
9. A flower freshness-preserving agent comprising the auxin and anthranilic acid activity inhibitor according to any one of 1 to 3.

The present invention provides an active inhibitor that inhibits both auxin and anthranilic acid, and a novel plant growth regulator containing the same.
When used on a plant during the growing season, the plant growth regulator of the present invention suppresses the growth effect of auxin on the plant, promotes tap root elongation, suppresses adventitious root elongation, suppresses sprouting, and pollen formation. It exerts effects such as suppressing the growth of lateral buds and suppressing the growth of the above-ground parts of the plant. It also suppresses the action of endogenous auxin in the plant body, suppresses overelongation during seedling raising, and promotes the formation of branches.
Furthermore, the plant growth regulator of the present invention does not cause plant death, which occurs with auxin inhibitors, when administered at a concentration that suppresses the growth of lateral roots.
The plant growth regulator of the present invention promotes the development of branches and extends the flowering period in mature plants.
Additionally, when used for flowers, the plant growth regulator of the present invention can delay the wilting of flowers and maintain the freshness of the flowers for a long period of time.

1 is a graph showing the results of an adzuki bean rooting test of Compound 1 (AAA12). 1 is a graph showing the test results of Compound 2 (AAA1) against indole acetic acid (IAA) in an adzuki bean rooting test. It is a graph showing the test results of Compound 2 (AAA1) against anthranilic acid in the adzuki bean rooting test. 1 is a graph of the results of an adzuki bean rooting test of Compound 3 (AAA1-me). It is a graph of the adzuki bean rooting test results of compound 4 (AAA11). It is a graph of the adzuki bean rooting test results of compound 5 (AAA18). It is a graph of the adzuki bean rooting test results of compound 6 (AAA23). It is a graph of the adzuki bean rooting test results of Compound 7 (AAA24). It is a graph of the adzuki bean rooting test results of compound 8 (AAA25). It is a graph of the adzuki bean rooting test results of compound 9 (AAA19). It is a graph of the adzuki bean rooting test results of compound 10 (AAA14). It is a graph of the adzuki bean rooting test results of Compound 11 (AAA30). It is a graph of the adzuki bean rooting test results of compound 12 (AAA36). It is a graph of the adzuki bean rooting test results of compound 13 (AAA34). It is a graph of the adzuki bean rooting test results of compound 14 (AAA38). It is a graph of the adzuki bean rooting test results of Compound 101 (Comparative Example 1). It is a graph of the adzuki bean rooting test results of Compound 102 (Comparative Example 2). It is a graph of the adzuki bean rooting test results of Compound 103 (Comparative Example 3). It is a graph of the adzuki bean rooting test results of Compound 104 (Comparative Example 4). It is an observation image of the taproot length 7 days after sowing in a taproot elongation inhibition test using Arabidopsis. It is a graph of the taproot length measurement results of the taproot elongation inhibition test using Arabidopsis. It is an observation image of the state of taproot development 11 days after sowing in a taproot elongation inhibition test using Arabidopsis. It is a graph showing the results of taproot length measurement 11 days after sowing in a taproot elongation inhibition test using Arabidopsis. It is a graph showing the measurement results of lateral root number density 11 days after sowing in a taproot elongation inhibition test using Arabidopsis. These are an observation image of lateral root density in test medium 1 and an observation image in test medium 2 in a test for endogenous auxin. It is a graph of measurement results of lateral root density in a test for endogenous auxin. This is an observation image of an effect confirmation test based on changes in the expression pattern of auxin-responsive genes. 1 is a graph of the results of an effect test of Compound 2 on lateral root formation of naphthalene acetic acid. It is a graph showing the results of a growth suppression effect test on broccoli seedlings. This is an observation image of a growth suppression effect test on broccoli seedlings. It is a graph showing the results of a branch development effect test on peas. These are the results of measuring the number of flowerings in a pot carnation flower flowering period extension test. It is a cumulative graph of the number of wilted flowers over time in a flowering period extension test of pot carnation flowers. It is a graph of the total number of wilted flowers in a flowering period extension test of pot carnation flowers. FIG. 1 shows the synthesis reaction of compound 15. FIG. 2 shows the synthesis reaction of compound 15. FIG. 3 shows the synthesis reaction of compound 15. FIG. 4 shows the synthesis reaction of compound 15. It is a graph showing the results of taproot length measurement 15 days after sowing in a taproot elongation inhibition test using Arabidopsis. It is a graph showing the results of a growth suppression effect test on broccoli seedlings.

In the process of searching for compounds that have an inhibitory effect on auxin and anthranilic acid, the present inventors discovered that the 4th position of the benzene ring in the anthranilic acid skeleton has a chemical structure in which the carbon skeletons are coplanar. They also discovered that a substance in which aromatic hydrocarbons are bonded with appropriate intermolecular distance has both anthranilic acid inhibitory effect and auxin inhibitory effect. It has been found that a substance having both of these inhibitory effects is useful as a plant growth regulator.
The term "plant growth regulator" as used in the present invention refers to a drug that has a regulating effect on the growth and development of plants by directly treating the plant with the agent, and is used to regulate the growth of agricultural crops.

The compound of the present invention that has an inhibitory effect on auxin and anthranilic acid and serves as a plant growth regulator is the following compound represented by Formula 1.

(However, A is a single bond via a -CH(=O)-NH- or -CH=CH- structure, R1 is a hydrogen atom, a halogen atom, or an alkyl group, and R2 is a hydrogen atom, A halogen atom, an alkyl group having 1 to 6 carbon atoms, R3 is a hydrogen atom, an alkyl group, or a halogen atom, R4 is OH or OCH3, and R5 is an amino group, a halogen atom, a hydroxyl group, or hydrogen (R1 and R2 may form a closed benzene ring.)
For convenience of explanation, the structure on the left side of A will be referred to herein as a "left ring structure" and the structure on the right side will be referred to as a "right ring structure".

In this compound, the structure of A connecting the left ring structure and the right ring structure is -CH(=O)-NH- or CH=CH structure, so that the left ring structure - A - right ring structure via A The chemical configurations of the carbon skeletons of are all arranged on the same plane (however, R4 is excluded from the configuration). Due to this arrangement, the compound exhibits inhibitory effects on both auxin and anthranilic acid.
The present inventors searched for a large number of compounds and found that the compound represented by the above formula 1 inhibits both the effects of auxin and anthranilic acid.

In addition, in the chemical structure of the compound of formula 1, when A is -CH(=O)-NH-, it has a double bond property and therefore takes a rigid, planar structure. This planar structure is called an "amide plane" (see Non-Patent Document 7). It is known that this occurs because the p-orbital of the nitrogen in the amide structure overlaps with the p-orbital of the carbonyl group (see Non-Patent Document 7). Therefore, when A is -CH(=O)-NH-, it also overlaps with the p orbital of the benzene ring/naphthalene ring adjacent to A, so the entire carbon skeleton other than the alkyl ester of R4 of the compound is on the same plane. Placed. Further, the order of the NH- and -CH(=O)- bonds in -CH(=O)-NH may be reversed.
This ability to change the structure of a molecule without changing its overall biological activity is known as bioequivalence (bioisostere). Furthermore, an example in which the same activity is maintained even if the order of -NH- and -CH(=O)- in the -CH(=O)-NH- structure is reversed is well known as amide isoster. (See Non-Patent Document 8).
It has been revealed that the compound used in the plant growth regulator of the present invention similarly maintains the same activity even if the order of the amino group and carbonyl group in the -CH(=O)-NH- structure is reversed.

Furthermore, it is known that even if the above-mentioned -CH(=O)-NH- structure is replaced with a -CH=CH- structure, the biological activity of the compound before substitution is similarly maintained (Non-Patent Document 8 reference). Similarly, even when the -CH(=O)-NH- structure of the compound used in the plant growth regulator of the present invention is replaced with the -CH=CH- structure, the inhibitory effect on auxin and anthranilic acid is maintained. It was revealed.
In this case, as in the -CH(=O)-NH- structure, the p orbital of the -CH=CH- structure also overlaps with the p orbital of the adjacent benzene ring/naphthalene ring, so the entire carbon skeleton other than R4 of the compound are placed on the same plane.
On the other hand, even if the carbon skeletons are on the same plane, 4-benzoylbenzoic acid (4-benzoylbenzoic acid, formula 15), which has only one carbon between the left ring structure and the right ring structure, and the left ring structure and It was found that biphenyl-4-carboxylic acid (Formula 16), in which the right ring structure is directly bonded without an atom, exhibits neither auxin inhibitory effect nor anthranilic acid inhibitory effect. From this, it was found that an appropriate intermolecular distance is required between the left ring structure and the right ring structure to maintain activity.
Furthermore, even if the intermolecular distance between the left ring structure and the right ring structure is appropriate, 2-amino-4-[(cyclohexylamino)carbonyl]benzoic acid (2-amino -4- [(cyclohexylamino)carbonyl]benzoic a
cid, Formula 17), and 2-amino-4-[(4-tert-butylphenylamino)carbonyl]benzoic acid (2-amino-4-[(4-tert-butylphenylamino)carbonyl]benzoic acid ( It was found that 2-amino-4-[(4-tert-butylphenylamino)carbonyl]benzoic acid, formula 18) also exhibits neither auxin inhibitory effect nor anthranilic acid inhibitory effect. From this, it became clear that in order to maintain activity, it is necessary that the entire left ring structure and right ring structure have a planar structure.

The agent having an inhibitory effect on auxin and anthranilic acid of the present invention can be used as a plant growth regulator. The compounds represented by Formula 1, especially Compounds 1 to 15 shown in Examples, all have strong inhibitory effects on both auxin and anthranilic acid.
The auxin inhibitory effect can be evaluated by apical bud elongation inhibition test, adventitious root development inhibition test, etc. using a known substance such as indole acetic acid (IAA) as a positive target.
Furthermore, the inhibitory effect of anthranilic acid can be confirmed as anti-anthranilic acid activity by comparing the inhibitory effect on adventitious root development using anthranilic acid as a positive target and as an indicator. This anti-anthranilic acid activity can be evaluated by the method disclosed in Patent Document 6, in which the cut ends of adzuki bean seedlings are soaked and the development and elongation of adventitious roots are observed. The inhibitory effect of anthranilic acid is hereinafter referred to as anti-anthranilic acid activity in the present specification.

The compound represented by Formula 1 can be synthesized as necessary.
For example, a compound in which A is -CH(=O)-NH- structure and R4 is OCH3 can be synthesized using 1-methyl 2-aminoterephthalate, monomethyl terephthalate, or methyl 4-aminobenzoate as starting materials. , compounds having substituents corresponding to various R1 to R3 can be obtained.
Note that the compound in which R4 is OH can be synthesized by hydrolyzing the above-mentioned compound in which R4 is OCH3. As the hydrolysis method, alkaline hydrolysis, acid hydrolysis, enzyme treatment, etc. can be used. Alkaline hydrolysis is preferred because it is easy to operate.
A compound in which A has a -CH═CH- structure can be synthesized by cross-coupling trans-2-phenylvinylboronic acid and a benzoic acid derivative substituted with halogen at the 4-position by a conventional method.

After the synthesis reaction is complete, to collect the product from the reaction solution, distill off the reaction solvent, add a product-soluble organic solvent that does not mix with water and water, adjust the pH of the aqueous phase as appropriate, and then perform solvent extraction. After collecting and drying the organic solvent layer and distilling off the organic solvent, it may be recrystallized from a single solvent or a mixed solvent as required. Further, if necessary, isolation may be performed by a separation means such as high performance liquid chromatography (HPLC).

Below, the names, English names, and chemical formulas of typical compounds used as the plant growth regulator of the present invention are illustrated. Naturally, the present invention is not limited to these compounds.

Compound 1 (AAA12)
Symbols in parentheses are abbreviations added for convenience. The same applies to the compounds shown below. 4-[(phenylamino)carbonyl]-benzoic acid

Compound 2 (AAA1)
2-Amino-4-[(phenylamino)carbonyl]-benzoic acid

Compound 3 (AAA1-me)
Methyl-2-Amino-4-[(phenylamino)carbonyl]-benzoate

Compound 4 (AAA11)
2-Amino-4-[(1-naphthylamino)carbonyl]-benzoic acid

Compound 5 (AAA18)
2-Amino-4-[(2,3-dimethylphenylamino)carbonyl]-benzoic acid

Compound 6 (AAA23)
2-Amino-4-[(2-fluorophenylamino)carbonyl]-benzoic acid

Compound 7 (AAA24)
2-Amino-4-[(3-fluorophenylamino)carbonyl]-benzoic acid

Compound 8 (AAA25)
2-Amino-4-[(4-fluorophenylamino)carbonyl]-benzoic acid

Compound 9 (AAA19)
4-(benzoylamino)benzoic acid

Compound 10 (AAA14)
2-hydroxy-4-(benzoylamino)benzoic acid

Compound 11 (AAA30)
2-Chloro-4-(benzoylamino)benzoic acid

Compound 12 (AAA36)
4-(4-fluorobenzoylamino)benzoic acid

Compound 13 (AAA34)
4-Stilbenecarboxylic acid

Compound 14 (AAA38)
2-amino-4-(benzoylamino)benzoic acid

Compound 15 (AAA37)
2-Amino-4-styrylbenzoic acid

The auxin and anthranilic acid activity inhibitor of the present invention can also be used in combination of two or more compounds. It can also be used in combination with known auxin synthesis inhibitors.

The auxin and anthranilic acid activity inhibitor of the present invention can be used as a plant growth regulator, a lateral root development inhibitor, a main root elongation promoter, a plant overelongation inhibitor, a branching development promoter, and It can be used as a flower freshness preserving agent.
Further, it may be formulated using carriers used in plant growth regulators and agricultural chemicals commonly used in the agricultural field, such as hydrating powders, emulsions, granules, powders, and surfactants. For example, solid carriers include mineral powders (kaolin, bentonite, clay, montmorillonite, talc, diatomaceous earth, mica, vermiculite, gypsum, calcium carbonate, phosphorous lime, etc.), vegetable powders (soybean flour, wheat flour, wood flour, tobacco, etc.). Powder, starch, crystalline cellulose, etc.), polymer compounds (petroleum resin, polyvinyl alcohol resin, polyvinyl acetate resin, polyvinyl chloride, ketone resin, etc.), alumina, waxes, etc. can be used. Examples of liquid carriers include alcohols (methanol, ethanol, propanol, butanol, ethylene glycol, benzyl alcohol, etc.), aromatic hydrocarbons (toluene, benzene, xylene, etc.), chlorinated hydrocarbons (chloroform, carbon tetrachloride, monochlorobenzene, etc.), ethers (dioxane, tetrahydrofuran, etc.), ketones (acetone, methyl ethyl ketone, etc.), esters (ethyl acetate, butyl acetate, etc.), acid amides (N,N-dimethylacetamide, etc.) , ether alcohols (such as ethylene glycol ethyl ether), or water can be used.

As the surfactant used for the purpose of emulsification, dispersion, diffusion, etc., any of nonionic, anionic, cationic, and amphoteric surfactants can be used. Examples of surfactants that can be used in the present invention include polyoxyethylene alkyl ether, polyoxyethylene alkylaryl ether, polyoxyethylene fatty acid ester, sorbitan fatty acid ester, polyoxyethylene sorbitan fatty acid ester, oxyethylene polymer , oxypropylene polymer, polyoxyethylene alkyl phosphate ester, fatty acid salt, alkyl sulfate ester salt, alkyl sulfonate, alkylaryl sulfonate, alkyl phosphate, alkyl phosphate ester salt, polyoxyethylene alkyl sulfate ester, These include quaternary ammonium salts, oxyalkylamines, lecithins, and saponins. Moreover, gelatin, casein, sodium alginate, starch, agar, polyvinyl alcohol, etc. can be used as adjuvants, if necessary.

The auxin and anthranilic acid activity inhibitor, plant growth regulator, lateral root development inhibitor, tap root elongation promoter, plant overelongation inhibitor, branching development promoter, and flower freshness preserving agent of the present invention are in the form of a preparation. There is no limit to the formulation, and it can be formed into any formulation such as powder, granules, granules, wettable powders, flowables, emulsions, and pastes. It can be produced by mixing, stirring, spray drying, etc. other components according to conventional methods.
When applied to plants, it can be used as a soil treatment agent, a foliar treatment agent, a seed treatment agent before sowing, a treatment agent for plants before transplantation, a treatment agent for plants at the time of transplantation, etc. Furthermore, in hydroponic culture, it may be used by mixing it with a hydroponic solution, and in tissue culture, it may be used by suspending or dissolving it in a medium.
The auxin and anthranilic acid activity inhibitor, plant growth regulator, lateral root development inhibitor, tap root elongation promoter, plant overelongation inhibitor, branching development promoter, and flower freshness preserving agent of the present invention are suitable for the adventitious roots of plants. Suppresses emergence and promotes tap root growth. Furthermore, when applied to a target plant, the tap root will elongate, the yield of accumulated sugars and starches will increase, branching will be promoted, and the yield will increase. Moreover, when applied during seedling raising, softening of the seedlings can be suppressed and strong seedlings for transplantation can be provided. Furthermore, if applied during storage of potatoes, sweet potatoes, etc., rooting and germination will be inhibited, allowing long-term storage.

The auxin and anthranilic acid activity inhibitor, plant growth regulator, lateral root development inhibitor, tap root elongation promoter, plant overelongation inhibitor, branching development promoter, and flower freshness preserving agent of the present invention are used for spraying. The concentration used in this case can be preferably in the range of 0.01 to 10,000 ppm, more preferably 1 to 5,000 ppm, particularly preferably 5 to 1,000 ppm of the compound of formula 1. Particularly when used for seedlings during the seedling-raising stage, it is desirable to spray 50 to 200 ml of the diluted solution at the above concentration per liter of culture soil. In this case, a spreading agent may be used, and the type and amount of the spreading agent used are not particularly limited. It is also effective when sprayed on the leaves.
When directly applied to soil, including when mixed with fertilizer, the amount used is preferably 100 to 10,000 g, particularly 500 to 5,000 g per hectare. In particular, when used for seedlings during the growing stage, it is desirable to use 0.001 to 10 g per liter of culture soil. In this case, it may be mixed in advance with the culture soil before sowing, or it may be sprayed during the seedling-raising period.

When used for seed treatment before sowing, use water, alcohols (methanol, ethanol, etc.), ketones (acetone, etc.), aromatic hydrocarbons (toluene, benzene, etc.), chlorinated hydrocarbons (chloroform, chloride, etc.). dilute with a liquid carrier such as methylene (such as methylene), ethers (such as diethyl ether), or esters (such as ethyl acetate) to a concentration of 0.01 to 100,000 ppm, and spray onto dried seeds, or immerse the dried seeds in the diluted solution. It can also be absorbed into seeds. The immersion time is not particularly limited, but is preferably 1 second to 120 minutes. Furthermore, the liquid carrier may be evaporated from the treated seeds by air drying, vacuum drying, heat drying, vacuum drying, or the like. It is also possible to use a formulation prepared using a solid carrier of mineral powder such as clay and adhered to the seed surface. It can also be mixed with commonly used seed coating agents and seed coating films to coat seeds.
When used in tissue culture or cell culture, the compound of formula 1 is added to a commonly used plant tissue culture medium (MS medium, White medium, Gamborg's B5 medium, etc.) at a concentration of preferably 0.01 to 0.01. It can be used dissolved or suspended at a concentration of 10,000 ppm, particularly preferably in the range of 0.1 to 1,000 ppm.
When directly absorbed into plants before transplantation, the roots or the entire plant can be immersed in a solution in which the compound of formula 1 is diluted or suspended to a concentration of 0.1 to 1000 ppm.

The timing of administration of the auxin and anthranilic acid activity inhibitor, plant growth regulator, lateral root development inhibitor, tap root elongation promoter, plant overelongation inhibitor, branching development promoter, and flower freshness preserving agent of the present invention is as follows: It can be used at any time during the growing period, but it should be sprayed as appropriate while checking the growth situation. Normally, if the seedlings grow too long, they are sprayed directly onto the plants to suppress this growth. Alternatively, it can be sprayed as appropriate during the storage period of the rhizomes after harvesting to suppress germination and rooting.
When spraying for the purpose of weeding, it can be sprayed at any appropriate time when weeding is required.

The auxin and anthranilic acid activity inhibitor, plant growth regulator, lateral root development inhibitor, tap root elongation promoter, plant overelongation inhibitor, branching development promoter, and flower freshness preserving agent of the present invention can be applied. Examples of plants include, but are not limited to, solanaceae such as tomatoes, green peppers, chili peppers, and eggplants; cucurbits such as cucumbers, pumpkins, melons, and watermelons; raw and spicy vegetables such as celery, parsley, and lettuce; and green onions. , green onions such as onions and garlic, legumes such as soybeans, groundnuts, green beans, peas, and azuki beans, other fruit vegetables such as strawberries, tap roots such as radish, turnips, carrots, and burdock, taro, cassava, potato, sweet potato, Potatoes such as Japanese yam, soft vegetables such as asparagus, spinach, honeysuckle, flowers such as lisianthus, stock, carnations, chrysanthemums, grains such as rice and corn, bentgrass, grass such as orifolia, rapeseed, sunflowers, etc. oil crops, sugar crops such as sugar cane and sugar beets, fiber crops such as cotton and rushes, forage crops such as clover, sorghum, and dent corn, deciduous fruit trees such as apples, pears, grapes, and peaches; Examples include citrus fruits such as mandarin orange, lemon, and grapefruit, and woody fruits such as azalea, azalea, and cedar. Among these, plants such as tomatoes, bell peppers, chili peppers, eggplants, cucumbers, pumpkins, melons, watermelons, celery, parsley, lettuce, green onions, onions, asparagus, lisianthus, stock, rice, bentgrass, oriole, sugar beets, rushes, etc. It is particularly effective against chrysanthemums, carnations, azaleas, azaleas, and grapes. In addition, examples of plants of the Poaceae family for the purpose of lodging prevention include rice, wheat, barley, rye, and corn.

Furthermore, for the purpose of improving the effects of the present invention, it can be used in combination with other plant growth regulators (auxin synthesis inhibitors) as described above, and a synergistic effect can be expected in some cases.
For example, when raising seedlings under conditions such as high planting densities, high humidity, and lack of sunlight, which tend to make them grow easily, anti-gibberellins (paclobutrazol, uniconazole, P, ancymidol, etc.), growth inhibitors (daminozid, etc.), and ethylene generators (ethephon, etc.) may be used in combination.

The plant growth regulator, lateral root development inhibitor, tap root elongation promoter, plant overelongation inhibitor, branching development promoter, and flower freshness preserving agent of the present invention include various insecticides, fungicides, microbial pesticides, fertilizers, etc. It is also possible to mix or use with.

1. Production example <Compound 1 (AAA12)>
5.0 g of monomethyl terephthalate was dissolved in 50 mL of methylene chloride solution to obtain a solution.
5.14 g of aniline, 5.33 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 3.88 ml of triethylamine, and 0.38 g of 1-hydroxybenzotriazole were dissolved in 200 mL of methylene chloride solution to form a solution. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 0.2 g of the obtained compound, 50 mL of 50% ethanol, and 5 mL of 1N sodium hydroxide were sequentially added to the container, and the mixture was stirred at 65° C. for 1 hour. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were washed with acetone and methanol and dried under reduced pressure to obtain 0.12 g (yield 1.8%) of the compound. I got it. The mass spectrum and melting point of this compound were measured, and the structure of Formula 5 was confirmed.
Proton NMR, mass spectrum, and melting point of this compound were measured, and the structure of Formula 2 was confirmed.

<Compound 2 (AAA1)>
5.0 g of 1-methyl 2-aminoterephthalate was dissolved in 50 mL of methylene chloride solution to form a solution. 10 ml of aniline, 4.9 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 10.7 ml of triethylamine, and 0.3 g of 1-hydroxybenzotriazole were dissolved in 200 ml of methylene chloride solution to form a solution. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals (Compound 2) were recrystallized from acetone and then dried under reduced pressure. 3 g of the obtained compound, 50 mL of 50% ethanol, and 5 mL of 1N sodium hydroxide were sequentially added to the container, and the mixture was stirred at 65° C. for 1 hour. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from acetone and dried under reduced pressure to obtain 0.36 g (yield 12.9%) of the compound. I got it.
Proton NMR, mass spectrum, and melting point of this compound were measured, and the structure of Formula 3 was confirmed.

<Compound 3 (AAA1-me)>
5.0 g of 1-methyl 2-aminoterephthalate was dissolved in 50 mL of methylene chloride solution to form a solution. 10 ml of aniline, 4.9 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 10.7 ml of triethylamine, and 0.3 g of 1-hydroxybenzotriazole were dissolved in 200 ml of methylene chloride solution to form a solution. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium bicarbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and methylene chloride was distilled off under reduced pressure to give 3.489 g (50.4%) of The compound was obtained. The mass spectrum and melting point of this compound were measured, and the structure of Formula 4 was confirmed.

<Compound 4 (AAA11)>
5.0 g of 1-methyl 2-aminoterephthalate was dissolved in 50 mL of methylene chloride solution to form a solution. Dissolve 7.16 g of 1-naphthylamine, 4.9 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 3.58 ml of triethylamine, and 0.35 g of 1-hydroxybenzotriazole in 200 mL of methylene chloride solution to obtain a solution. did. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 1 g of the obtained compound, 50 mL of 50% ethanol, and 5 mL of 1N sodium hydroxide were sequentially added to a container, and the mixture was stirred at 65° C. for 1.5 hours. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from acetone and dried under reduced pressure to obtain 0.33 g (yield 4.2%) of the compound. I got it.
Proton NMR, mass spectrum, and melting point of this compound were measured, and the structure of Formula 5 was confirmed.

<Compound 5 (AAA18)>
5.0 g of 1-methyl 2-aminoterephthalate was dissolved in 25 mL of methylene chloride solution to form a solution. Dissolve 6.51 g of 2,3-dimethylamine, 4.9 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 3.58 ml of triethylamine, and 0.35 g of 1-hydroxybenzotriazole in 125 mL of methylene chloride solution. It was made into a solution. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from a mixture of acetone and methanol and then dried under reduced pressure. 2.1 g of the obtained compound, 100 mL of 50% ethanol, and 10 mL of 1N sodium hydroxide were sequentially added to a container, and the mixture was stirred at 65° C. for 1 hour. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from acetone and dried under reduced pressure to obtain 0.30 g (yield 14.9%) of the compound. I got it.
Proton NMR, mass spectrum, and melting point of this compound were measured, and the structure of Formula 6 was confirmed.

<Compound 6 (AAA23)>
5.0 g of 1-methyl 2-aminoterephthalate was dissolved in 50 mL of methylene chloride solution to form a solution. Dissolve 5.97 g of 2-fluoroaniline, 4.9 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 3.58 ml of triethylamine, and 0.35 g of 1-hydroxybenzotriazole in 200 mL of methylene chloride solution. And so. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 0.57 g of the obtained compound, 50 mL of 50% ethanol, and 5 mL of 1N sodium hydroxide were sequentially added to the container, and the mixture was stirred at 65° C. for 1 hour. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from acetone and dried under reduced pressure to obtain a compound of 0.23 (yield 3.3%). I got it.
Proton NMR, mass spectrum, and melting point of this compound were measured, and the structure of Formula 7 was confirmed.

<Compound 7 (AAA24)>
5.0 g of 1-methyl 2-aminoterephthalate was dissolved in 50 mL of methylene chloride solution to form a solution. Dissolve 5.97 g of 3-fluoroaniline, 4.9 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 3.58 ml of triethylamine, and 0.35 g of 1-hydroxybenzotriazole in 200 mL of methylene chloride solution. And so. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 0.27 g of the obtained compound, 100 mL of 50% ethanol, and 10 mL of 1N sodium hydroxide were sequentially added to the container, and the mixture was stirred at 65° C. for 1 hour. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from acetone and dried under reduced pressure to obtain 0.15 g (yield 2.1%) of the compound. I got it.
The mass spectrum and melting point of this compound were measured, and the structure of Formula 8 was confirmed.

<Compound 8 (AAA25)>
5.0 g of 1-methyl 2-aminoterephthalate was dissolved in 50 mL of methylene chloride solution to form a solution. Dissolve 5.97 g of 4-fluoroaniline, 4.9 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 3.58 ml of triethylamine, and 0.35 g of 1-hydroxybenzotriazole in 200 mL of methylene chloride solution. And so. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 0.17 g of the obtained compound, 100 mL of 50% ethanol, and 10 mL of 1N sodium hydroxide were sequentially added to the container, and the mixture was stirred at 65° C. for 1 hour. After adding concentrated hydrochloric acid to the reaction solution to adjust the pH to 8.0 and extracting with ethyl acetate, concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted three times with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from acetone and dried under reduced pressure to obtain 0.08 g (yield 1.2%) of the compound. I got it.
Proton NMR, mass spectrum, and melting point of this compound were measured, and the structure of Formula 9 was confirmed.

<Compound 9 (AAA19)>
5.0 g of methyl 4-aminobenzoate was dissolved in 50 mL of methylene chloride solution to form a solution. 4.0 g of benzoic acid, 6.9 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 5 ml of triethylamine, and 0.5 g of 1-hydroxybenzotriazole were dissolved in 100 mL of methylene chloride solution to form a solution. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 1.0 g of the obtained compound, 100 mL of 50% ethanol, and 10 mL of 1N sodium hydroxide were sequentially added to a container, and the mixture was stirred at 65° C. for 1 hour. After adding concentrated hydrochloric acid to the reaction solution to adjust the pH to 8.0 and extracting with ethyl acetate, concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted three times with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from a mixture of acetone and methanol and dried under reduced pressure to give 0.41 g (yield 5. 1%) of the compound was obtained.
Proton NMR, mass spectrum, and melting point of this compound were measured, and the structure of Formula 10 was confirmed.

<Compound 10 (AAA14)>
5.0 g of methyl 4-aminosalicylate was dissolved in 50 mL of methylene chloride solution to form a solution. 3.65 g of benzoic acid, 6.2 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 4.5 ml of triethylamine, and 0.4 g of 1-hydroxybenzotriazole were dissolved in 200 mL of methylene chloride solution to prepare a solution. . The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 0.7 g of the obtained compound, 50 mL of 50% ethanol, and 6 mL of 1N sodium hydroxide were sequentially added to the container, and the mixture was stirred at 65° C. for 1 hour. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from acetone and dried under reduced pressure to obtain 0.25 g (yield 3.3%) of the compound. I got it.
The mass spectrum and melting point of this compound were measured, and the structure of Formula 11 was confirmed.

<Compound 11 (AAA30)>
5 g of 4-amino-2-chlorobenzoic acid was dissolved in 15 ml of methanol, 20 ml of trimethylsilyldiazomethane (about 10% hexane solution, about 0.6 mol/L) was added dropwise, and the mixture was stirred at room temperature overnight. The reaction solution was scaled up with ethyl acetate, washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then ethyl acetate was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 0.56 g of the obtained compound, 0.41 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 0.3 ml of triethylamine, and 0.3 g of 1-hydroxybenzotriazole were dissolved in 20 mL of methylene chloride solution. And so. 0.23 g of benzoic acid was dissolved in 5 ml of methylene chloride. The latter solution was added to the former solution under ice cooling and stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 0.19 g of the obtained compound, 20 mL of 50% ethanol, and 5 mL of 1N sodium hydroxide were sequentially added to the container, and the mixture was stirred at 60° C. for 3 hours. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from methanol and dried under reduced pressure to obtain 0.1 g (yield 1.3%) of the compound. I got it.
The mass spectrum and melting point of this compound were measured, and the structure of Formula 12 was confirmed.

<Compound 12 (AAA36)>
5.0 g of methyl 4-aminobenzoate was dissolved in 50 mL of methylene chloride solution to form a solution. Dissolve 4.6 g of 4-fluorobenzoic acid, 6.86 g of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, 5 ml of triethylamine, and 0.5 g of 1-hydroxybenzotriazole in 200 mL of methylene chloride solution to obtain a solution. did. The first solution was added to the second solution under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution was washed successively with water, 5% aqueous sodium hydrogen carbonate solution, 1N hydrochloric acid, and water, dried over anhydrous magnesium sulfate, and then methylene chloride was distilled off under reduced pressure. The obtained crystals were recrystallized from acetone and then dried under reduced pressure. 1 g of the obtained compound, 50 mL of 50% ethanol, and 5 mL of 1N sodium hydroxide were sequentially added to a container, and the mixture was stirred at 65° C. for 1 hour. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted with ethyl acetate. Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from a mixture of acetone and methanol and dried under reduced pressure to give 0.21 g (yield 2. 5%) of the compound was obtained.
The mass spectrum and melting point of this compound were measured, and the structure of Formula 13 was confirmed.

<Compound 13 (AAA34)>
5 g of ethyl stilbene-4-carboxylate, 50 mL of 50% ethanol, and 10 mL of 1N sodium hydroxide were sequentially added to the container, and the mixture was stirred at 65° C. for 6 hours. Concentrated hydrochloric acid was added to the reaction solution to adjust the pH to 8.0, and the mixture was extracted twice with ethyl acetate.Concentrated hydrochloric acid was added to the aqueous phase to adjust the pH to 2.5, and the mixture was extracted twice with ethyl acetate. After drying the ethyl acetate phase over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the obtained crystals were recrystallized from a mixture of acetone and methanol and dried under reduced pressure to give 0.16 g (yield: 3. 3%) of the compound was obtained.
The mass spectrum and melting point of this compound were measured, and the structure of Formula 14 was confirmed.

<Compound 14 (AAA38)>
0.5 g of 4-amino-2-nitrobenzoic acid, 25 mL of methanol, and 25 mL of trimethylsilyldiazomethane were added, and the mixture was stirred at room temperature for 2 hours. After adjusting the reaction solution to pH 8.0, 0.3 g of the compound obtained by drying the fraction extracted with ethyl acetate was dissolved in 40 ml of dichloromethane, and 0.27 mL of pyridine was added. The reaction solution was ice-cooled, 0.3 mL of benzoyl chloride mixed with 40 mL of dichloromethane was added, and the mixture was stirred for 20 hours while returning to room temperature. The reaction solution was washed with water, 5% aqueous sodium hydrogen carbonate solution, 1N acetic acid, and water, dehydrated over anhydrous magnesium sulfate, and then dried. The crystals were dissolved in n-hexane containing 20% ethyl acetate, and 2
It was adsorbed on a silica gel (Wako Gel C-100) column prepared with n-hexane containing 0% ethyl acetate, and what was eluted with n-hexane containing 50% ethyl acetate was dried, washed with diethyl ether, and dried to give 160 mg of crystals. I got it. The crystals were dissolved in ethyl acetate under an argon atmosphere, and 19.2 g of 10% palladium on carbon was added. The inside of the reaction vessel was replaced with hydrogen gas, and after stirring for 4 hours, 34.6 mg of 10% palladium on carbon was further added and stirred for 4 hours. The inside of the reaction vessel was replaced with argon, and after filtering through Celite, the solvent was distilled off under reduced pressure. 110.6 mg of the obtained crystals were dissolved in 15 mL of 50% ethanol and heated to 65°C. 1 mL of 1N sodium hydroxide was added to this, and the mixture was stirred for 3 hours. The reaction solution was washed with ethyl acetate, adjusted to pH 2.5 and extracted with ethyl acetate, and the ethyl acetate layer was separated and collected, dried over anhydrous magnesium sulfate, and dried under reduced pressure. Solidification gave 74 mg (10.5%) of compound 14.
The mass spectrum and melting point of this compound were measured, and the structure of Formula 15 was confirmed.

<Compound 15 (AAA37)>
Synthesis of 2-amino-4-styrylbenzoic acid (compound 15) (step 1)
The reaction in step 1 is shown in FIG.
After adding a 2.64 M hexane solution (0.85 mL) of n-BuLi to 2,2,6,6-tetramethylpiperidine (0.374 mL) at -78°C, the temperature was raised until the tetramethylpiperidine was dissolved. The mixture was again cooled to -78°C and stirred for 10 minutes. A 1.09M hexane solution (1.93 mL) of diethylzinc was added, and the temperature was raised to room temperature over 50 minutes. The resulting mixture was diluted with THF (1.6 mL) to prepare a 0.5M hexane-THF solution of LiTMP0.1 [ZnEt2(TMP)].
A 0.5 M hexane-THF solution of LiTMP0.1 [ZnEt2 (TMP)] (3
．． 0 mL) and stirred for 1 hour. After cooling on ice, a THF solution (1.6 mL) of copper(II) 2-ethylhexanoate (35.0 mg) was added. The obtained mixture was added to solvent-free Compound (R1) (838.0 mg) under ice cooling, and the mixture was stirred at room temperature overnight.
After adding 2-propanol to the resulting mixture, it was filtered through alumina, and the resulting filtrate was concentrated under reduced pressure. The obtained residue was subjected to silica gel column chromatography (hexane:ethyl acetate=49:1) to obtain 142.7 mg of intermediate IM-2 represented by the above formula in a yield of 58%.
The 1H-NMR data of the obtained intermediate IM-2 is shown below.
1H-NMR (CDCl3, 500.16MHz): δ 7.59 (d, 1H, J = 8.0 Hz, Ar-H), 7.55 (d, 2H, J = 6.9 Hz, 2H, Ar-H), 7.41 (t, 2H , J = 8.0 Hz, Ar-H), 7.35-7.33 (m, 2H, Ar-H), 7.31 (s, 1H, Ar-H), 7.24 (d, 1H, J = 15.8 Hz, =CH-) , 7.07 (d, 1H, J = 16.0 Hz, =CH-)

(Step 2)
The reaction in step 2 is shown in FIG.
A 1.03M hexane solution of DIBAL-H (5.7 mL) was added to a mixture of intermediate IM-2 (71.4 mg) and dichloromethane (2.9 mL) at -78°C, and the mixture was heated to -30°C over 30 minutes. The temperature rose to .
1M hydrochloric acid was added to the resulting mixture, stirred at room temperature for 1 hour, and then extracted with dichloromethane. The obtained organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure.
Tert-butyl alcohol (2.18 mL), water (0.73 mL), THF (1.5 mL), 2-methyl-2-butene (0.31 mL) and dihydrogen phosphate were added to the concentrated residue containing intermediate IM-3. Sodium dihydrate (113.0 mg) and sodium chlorite (108.0 mg) were added, and the mixture was stirred at room temperature for 40 minutes.
Saturated brine was added to the resulting mixture, and the mixture was extracted with ethyl acetate. The obtained organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure.
Acetonitrile (2.16 mL) and sodium iodide (48.0 mg) were added to the concentrated residue containing intermediate IM-4, and the mixture was stirred for 5 minutes. A 3M solution of trimethylsilyl chloride in acetonitrile (0.11 mL) was added and stirred at room temperature for 75 minutes.
A 10% aqueous sodium thiosulfate solution was added to the resulting mixture, and the mixture was extracted with ethyl acetate. The obtained organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained residue was recrystallized in hot toluene to obtain 12.4 mg of Compound A with a yield of 16%.
Proton NMR was measured for the obtained compound.

A compound in which A has a -CH═CH- structure can be synthesized by cross-coupling trans-2-phenylvinylboronic acid and a benzoic acid derivative substituted with halogen at the 4-position by a conventional method.
In addition, for example, a compound having an amino group at R5 and a -CH=CH- structure in A is hydrolyzed to form an imine by DIBIAL-H reduction from a nitrile intermediate synthesized from cyanostilbene using ortho-azidation. It can be synthesized by oxidizing the intermediate aldehyde produced in this manner to derive a carboxylic acid, and finally reducing the azide group (see Figure 37).
The compound of formula (IM-2) is prepared by azidating the cyanostilbene of formula (IM-1) at the ortho position using LiTMP0.1[ZnEt2(TMP)]/Cu(eh)2. can do.
In this case, the method described in Journal of American Chemical Society, Volume 139, Issue 33, Pages 11622-11628, 2017 can be referred to.

The compound of formula (IM-3) can be prepared by subjecting the nitrile of formula (IM-2) to DIBIAL-H reduction to form an imine and hydrolyzing the resulting imine. The compound of formula (A) can be prepared by oxidizing this aldehyde to derive a carboxylic acid and finally reducing the azide group. As for the oxidation conditions, it is preferable to use Pinnick-Claus oxidation, which has high functional group selectivity (see FIG. 38). Regarding the reduction of the azide group, reference can be made to the method described in Tetrahedron Letters, Volume 38, Issue 39, Pages 6945-6948, 1997.

The melting points, NMR, and mass spectrum values of Compounds 1 to 15 are shown in Table 1 below.

Compounds 1 to 15 can be prepared without using the above synthesis method. Moreover, commercially available compounds can also be used. Purification purity does not necessarily have to be a high purity product, but it can be used in the present invention.

2. Test example <Test to confirm the effect on adventitious root generation of cut ends of azuki beans soaked in auxin or anthranilic acid>
The anti-auxin and anti-anthranilic acid activities of Compounds 1 to 15 were evaluated using adventitious root development at the cut end of adzuki bean seedlings as an indicator.

(1) Test compounds Compounds 1 to 15 above and 4-benzoylbenzoic acid, which has a benzoyl group introduced at the 4-position of benzoic acid, as a compound with a similar chemical structural formula, in which part A is C(=O). (Compound 101, Comparative Example 1), biphenyl-4-carboxylic acid (Compound 102, Comparative Example 2), which has a structure in which A is directly connected without an atom, as a compound without the A part, and the carbon skeletons are on the same plane. 2-Amino-4-[(cyclohexylamino)carbonyl]-benzoic acid (Compound 103, AAA8, Comparative Example 3), 2-amino-4-[(4-tert-butylphenylamino)carbonyl ] -A test was conducted using benzoic acid (Compound 104, AAA9, Comparative Example 4).
The chemical formulas of Comparative Examples Compounds 101, 102, 103, and 104 are as follows.
<Comparative Example 1: Compound 101>
4-benzoylbenzoic acid

<Comparative Example 2: Compound 102>
biphenyl-4-carboxylic acid

<Comparative Example 3: Compound 103>
2-Amino-4-[(cyclohexylamino)carbonyl]-benzoic acid

<Comparative Example 4: Compound 104>
2-Amino-4-[(4-tert-butylphenylamino)carbonyl]-benzoic acid

Compound 103 is obtained by replacing the left ring of compound 2 with cyclohexane, and compound 104 is obtained by bonding tert-butyl to the left ring of compound 2, resulting in a compound in which the coplanarity of the carbon skeleton is lost. This is an example.

(2) Test method Each test compound was dissolved in an appropriate solvent and diluted with distilled water to prepare an aqueous solution with a final test concentration in the range of 10 μM to 1 mM. The pH of the solution was adjusted to pH 7, and adzuki bean adventitious root development promotion assay (Itagaki et al. 2003. Biological activities and structure-activity relationship of substitution compounds of N-[2-(3-indolyl)ethyl] succinamic
acid and N-[2-(1-naphthyl)ethyl] succinamic acid, derived from a new category of root-promoting substance, N-(phenethyl)succinamic acid analogs. Plant Soil 255: 67-75.).

In addition, the anti-auxin activity and anti-anthranilic acid activity of each test compound was determined in a test in which it was used in combination with the auxin indoleacetic acid (IAA) or atranilic acid (AntA). It was directly detected as
Note that the adzuki bean seedling sections used in the test were cut by a predetermined method, and the cut bases were immersed in each test solution for 48 hours. Then, the number of adventitious roots generated after 7 days was counted.
The number of repetitions of the test was 5 azuki seedlings per test.
Furthermore, when measuring the activity of each compound, the number of adventitious roots was similarly measured using adzuki bean seedlings treated with distilled water (DW) as a control. The measurement results were expressed as the total average value of the number of roots.

(3) Test results 1) Test results for compound 1 (AAA12) Figure 1 shows a graph of the test results.
Compound 1 (AAA12) completely inhibited rooting of azuki bean at a concentration of 0.1 mM. Rooting of a 10 μM solution of indole acetic acid or a 1 mM solution of anthranilic acid was also inhibited in a concentration-dependent manner.

2) Test results for compound 2 (AAA1) FIG. 2 shows a graph of the test results for compound 2 (AAA1) against indole acetic acid, and FIG. 3 shows a graph of the test results for anthranilic acid.
Compound 2 (AAA1) inhibited rooting in a 10 μM solution of indole acetic acid in a concentration-dependent manner (see FIG. 2). Note that the effect of indoleacetic acid was completely suppressed at a concentration of 0.3 mM. Compound 2 also inhibited rooting in a 0.3 mM anthranilic acid solution (FIG. 3).

3) Test results for compound 3 (AAA1-me) FIG. 4 shows a graph of the test results for compound 3 (AAA1-me).
Compound 3 (AAA1-me) inhibited rooting in a 10 μM solution of indole acetic acid or a 1 mM solution of anthranilic acid.

4) Test results for compound 4 (AAA11) FIG. 5 shows a graph of the test results for compound 4 (AAA11).
Compound 4 (AAA11) inhibited rooting in a 10 μM solution of indole acetic acid or a 1 mM solution of anthranilic acid in a concentration-dependent manner.

5) Test results for compound 5 (AAA18) FIG. 6 shows a graph of the test results for compound 5 (AAA18).
Compound 5 (AAA18) inhibited rooting in a 10 μM indoleacetic acid solution in a concentration-dependent manner. Compound 5 also inhibited rooting in a 1 mM anthranilic acid solution.

6) Test results for compound 6 (AAA23) FIG. 7 shows a graph of the test results for compound 6 (AAA23).
Compound 6 (AAA23) inhibited rooting in a 10 μM solution of indole acetic acid in a concentration-dependent manner. Furthermore, rooting was inhibited by 1 mM of anthranilic acid.

7) Test results for compound 7 (AAA24) FIG. 8 shows a graph of the test results for compound 7 (AAA24).
Compound 7 (AAA24) inhibited rooting in a 10 μM solution of indoleacetic acid and 1 mM anthranilic acid in a concentration-dependent manner.

8) Test results for compound 8 (AAA25) FIG. 9 shows a graph of the test results for compound 8 (AAA25).
Compound 8 (AAA25) inhibited rooting in a 10 μM solution of indoleacetic acid and 1 mM anthranilic acid in a concentration-dependent manner.

9) Test results for compound 9 (AAA19) FIG. 10 shows a graph of the test results for compound 9 (AAA19).
Compound 9 (AAA19) almost completely inhibited rooting in a 10 μM solution of indoleacetic acid and 1 mM anthranilic acid at a concentration of 0.1 mM.

10) Test results for compound 10 (AAA14) FIG. 11 shows a graph of the test results for compound 10 (AAA14).
Compound 10 (AAA14) almost completely inhibited rooting in a 10 μM solution of indoleacetic acid and 1 mM anthranilic acid at a concentration of 0.1 mM.

11) Test results for compound 11 (AAA30) FIG. 12 shows a graph of the test results for compound 11 (AAA30).
Compound 11 (AAA30) inhibited rooting in a 10 μM solution of indoleacetic acid and 1 mM anthranilic acid in a concentration-dependent manner.

12) Test results for compound 12 (AAA36) FIG. 13 shows a graph of the test results for compound 12 (AAA36).
Compound 12 (AAA36) inhibited rooting in a 10 μM solution of indoleacetic acid and 1 mM anthranilic acid in a concentration-dependent manner.

13) Test results for compound 13 (AAA34) FIG. 14 shows a graph of the test results for compound 13 (AAA34).
Compound 13 (AAA34) suppressed the rooting effect of a 10 μM solution of indole acetic acid in a concentration-dependent manner. Furthermore, rooting of 1 mM anthranilic acid was completely inhibited at a concentration of 0.1 mM.

14) Test results for compound 14 (AAA38) FIG. 15 shows a graph of the test results for compound 14 (AAA38).
Compound 14 (AAA38) inhibited rooting in distilled water at a concentration of 0.3 mM.

15) Test results for Compound 101 (Comparative Example 1) The test results for Compound 101 are shown in FIG. 16.
Compound 101 did not inhibit but enhanced the rooting effects of a 10 μM solution of indoleacetic acid and 1 mM anthranilic acid.

16) Test results for Compound 102 (Comparative Example 2) The test results for Compound 102 are shown in FIG. 17.
Compound 102 did not inhibit but enhanced the rooting effects of a 10 μM solution of indoleacetic acid and 1 mM anthranilic acid. Compound 102 at 0.3 mM killed the cut adzuki bean seedlings that were tested.

17) Test results for Compound 103 (Comparative Example 3) The test results for Compound 103 are shown in FIG. 18.
Compound 103 did not inhibit but enhanced the rooting effects of a 10 μM solution of indoleacetic acid and 1 mM of anthranilic acid.

18) Test results for Compound 104 (Comparative Example 4) The test results for Compound 104 are shown in FIG. 19.
Compound 104 did not inhibit the rooting effects of a 10 μM solution of indoleacetic acid and 1 mM of anthranilic acid. Compound 104 at 0.3 mM killed the cut adzuki bean seedlings that were tested.

The above test results and previous findings are summarized in Table 2 below.

As shown in Table 2, Compounds 1 to 15 have a carbon skeleton planar structure. It also has anti-auxin and anti-anthranilic acid effects. Further, compounds 1, 2, 4 to 14, which have a planar structure and the number of atoms in the skeleton of part A is 2, and compound 3, in which R4 is an alkyl bonded with an ester bond, exhibit anti-auxin and anti-anthranilic acid effects. On the other hand, compounds 101 and 102 (Comparative Examples 1 and 2) which have a similar structure but in which the A part is directly bonded without a carbonyl or an atom, and compounds 103 and 104 (Comparative Examples 3 and 4) which do not have a carbon skeleton planar structure, It did not show anti-auxin or anti-anthranilic acid effects. Therefore, the compound represented by Formula 1 has a planar carbon skeleton structure and has two intervening atoms in the A part, such as -CH(=O)-NH- or -CH=CH- (for example, - It was considered necessary to have a structure that can suppress the activity even when CH═CCH3- exists in order to exhibit both anti-auxin and anti-anthranilic acid effects.

(4) Consideration of test results From the test results, Compound 1 and Compound 8 and Compound 2 and Compound 14, in which the order of the amino group and carbonyl group of the amide bond are reversed, both exhibit anti-auxin activity. It was confirmed that the order of the amino group and carbonyl group does not matter.
Compounds 101 and 102 used as a control are compounds in which the left ring structure and the right ring structure have the same planar structure, but the binding mode of A is different. Compounds 101 and 102 did not exhibit anti-auxin activity or anti-anthranilic acid activity. Compounds 103 and 104, in which the carbon skeletons are not on the same plane, did not exhibit anti-auxin or anti-anthranilic acid activity.
On the other hand, Compound 3, in which R4 in Formula 1 is an ester-bonded alkyl, and the carbons constituting the alkyl group are not on the same plane, exhibited anti-auxin activity. The reason for this is that there are many types of esterases in plants (see Non-Patent Document 9), so by removing the ester structure of R4 in advance, it is assumed that it will not be involved in anti-auxin activity or anti-anthranilic acid activity. it was thought.
From the above, it is considered that for anti-auxin activity and anti-anthranilic acid activity, it is essential that the carbon skeletons other than the ester compound of R4 in Formula 1 be on the same plane.
In addition, 4-benzoylbenzoic acid (Compound 101) in which A in the structure of Formula 1 is carbonyl, and biphenyl-4-carboxylic acid (Biphenyl-4-carboxylic acid) in which A is directly bonded to A without an atom. 4-carboxylic acid) (Compound 102) did not show anti-auxin activity. From this, it was considered that A must have a -CH(=O)-NH- or -CH=CH- structure.

<Efficacy test of the compound of the present invention against exogenous auxin>
Next, Compound 2 (AAA1) was arbitrarily selected from the compounds of the present invention, and a growth regulation test against exogenous auxin was conducted using this compound.

Taproot elongation inhibition test 1 using Arabidopsis thaliana
(1) Test method Compound 2 (AAA1) of the present invention at concentrations of 10 μM, 20 μM, and 40 μM, and 1% sucrose-containing 1/2 MS solid medium (Nippon Pharmaceutical Co., Ltd.) containing indole acetic acid (IAA) at a concentration of 200 nM. ) was used as the test medium. This medium contains Arabidopsis wild type Col. -0 (Arabidopsis Biological Resource Center; Stock
CS7000) was sown and cultivated under long-day conditions (16 hours of light, 8 hours of darkness) and a temperature of 23°C.
A medium without addition of compound 2 and indole acetic acid (IAA) was set as a control (Cont), and a medium with only IAA added (IAA) was set as a control.
Next, 7 days after sowing, the tap root length of the germinated plants was measured. In addition, the main root length of each individual plant 11 days after sowing was measured in the same manner, and the number of lateral roots was also counted. The number of lateral roots was homogenized as the number per 1 mm of the main root length (number/mm).
The number of repetitions of the test was 8.

(2) Results 1) Taproot length a. 7 days after sowing. Taproot length Figure 20 shows an observation image of the taproot length 7 days after sowing. Further, FIG. 21 shows the results of measuring the tap root length.
The control (Cont) had a tap root length of about 25 mm. On the other hand, when 200 nM of IAA was added, taproot growth was significantly suppressed. When Compound 2 was added to this, the inhibition of taproot growth by IAA was resolved, and the taproot elongated in proportion to the amount of Compound 2 added (FIG. 21).

b. Rooting status As shown in FIG. 20, Cont showed normal rooting. On the other hand, when IAA was added, it was observed that the elongation of the main root was significantly hindered and lateral roots grew densely. Furthermore, when Compound 2 was added, the inhibition of taproot elongation was suppressed, and the same condition as Cont was observed.
From this observation result, it was confirmed that Compound 2 suppresses inhibition of taproot elongation through anti-auxin action.

2) Tap root length and number of lateral roots 11 days after sowing a. Taproot Length An observation image of the state of taproot development is shown in Fig. 22, a measurement result of the taproot length is shown in Fig. 23, and a lateral root number density is shown in Fig. 24.
As shown in FIG. 23, the taproot length exceeded 50 mm in Cont, but it only increased by 8 mm with IAA addition, which was almost the same as the measurement result on the 7th day. On the other hand, in the medium to which Compound 2 and IAA were added, the length was 40 mm when compound 2 was added at a concentration of 10 μM, 50 mm when 20 μM was added, and the length slightly exceeded 40 mm when 40 μM was added.

b. Lateral root density The lateral root density per 1 mm of taproot length is as shown in the image in Figure 22 and the graph of the measurement results in Figure 24. In the medium containing only IAA, the elongation of the taproot is suppressed and lateral roots develop and elongate. The density (number) of lateral roots per plant was extremely high. However, it was found that by adding Compound 2, it became comparable to Cont.

c. Observation of changes in the appearance of the rooting state As shown in Figure 22, from the images taken of the rooting state, it was observed that when only IAA was added to the medium, the taproot hardly elongated (see the second image from the left). ). On the other hand, it was observed that the main root and lateral roots were rooting in a favorable state by the addition of Compound 2.

From the above measurement and observation results of a, b, and c, Compound 2, that is, the compound according to the composition of the present invention, exhibits anti-auxin activity, grows the taproot after germination under auxin addition conditions, and grows the taproot along the taproot. It was found that the lateral roots were generated evenly and elongated further.

Taproot length suppression test 2 using Arabidopsis thaliana
(1) Test method Compound 15 (AAA37) of the present invention at concentrations of 10 μM, 20 μM, and 40 μM, and 1% sucrose-containing 1/2 MS solid medium (Nippon Pharmaceutical Co., Ltd.) containing indole acetic acid (IAA) at a concentration of 200 nM. ) was used as the test medium. This medium contains Arabidopsis wild type Col. -0 (Arabidopsis Biological Resource Center; Stock number CS7000) was sown and cultivated under long-day conditions (16 hours of light, 8 hours of darkness) and a temperature of 23°C. A medium without addition of compound 15 and indole acetic acid (IAA) was set as a control (Cont), and a medium with only IAA added (IAA) was set as a comparison control.
Next, 15 days after sowing, the taproot length of the germinated plants was measured. The number of repetitions of the test was 8.

(2) Results Figure 39 shows the results of measuring the length of the taproot on the 15th day of sowing. As shown in FIG. 39, the taproot length exceeded 70 mm in Cont, but it extended only 5 mm with IAA addition. On the other hand, in the medium supplemented with Compound 15 and IAA, the elongation was 30 mm at the addition concentration of Compound 15 of 0.04 mM and 50 mm at 0.1 mM.

<Suppression effect test on endogenous auxin>
Auxin (endogenous auxin) is produced within plants to control growth. The effect of compound 2 on this endogenous auxin was confirmed.
1. Test method A 1/2MS solid medium containing 1% sucrose (Nippon Pharmaceutical Co., Ltd.) containing compound 2 (AAA1) at four concentrations of 10 μM, 20 μM, and 40 μM was used as the test medium.
Separately, a medium containing 200 nM of IAA in addition to Compound 2 was prepared, and this was designated as Test Medium 2. This test medium 1 and test medium 2 contained Arabidopsis wild type Col. -0 (Arabidopsis Biological Resource Center) was sown and cultivated under long-day conditions (16 hours of light, 8 hours of darkness) at a temperature of 23°C. The number of lateral roots was measured 11 days after sowing. For the number of lateral roots, the measurement results were homogenized as the number per 1 mm of the main root length (number/mm).
The number of repetitions was 8.

2. Result a. Observation of Elongation of Taproot Length FIG. 25 shows an observation image of taproot growth when Compound 2 was added to Test Medium 1 (Cont) (upper row). Also shown are images observed when Compound 2 was added to Test Medium 2 (+IAA) (lower row).
In both cases, the taproot elongated smoothly.

b. Lateral Root Density Figure 26 shows a graph of lateral root density. As shown in test medium 1 (black bars), compound 2 reduces lateral root density in a concentration-dependent manner. This is considered to be due to the lack of endogenous auxin caused by the addition of Compound 2. On the other hand, in test medium 2 (white bar graph), it was found that the addition of indoleacetic acid increased the lateral root density compared to the addition of compound 2 alone. This was thought to be because the lack of endogenous auxin caused by Compound 2 was compensated for by the addition of IAA.

3. Discussion Compound 2, when used alone, suppresses the increase in lateral root density, as shown by the black bar graph. However, this effect of compound 2 is abolished by adding IAA (white bar graph). In other words, it was revealed that Compound 2 suppresses endogenous IAA, controls the growth of lateral roots, and regulates plant growth.

<Efficacy confirmation test of compound 2 based on changes in expression of auxin-responsive genes>
It is known that the Arabidopsis transformant promoter IAA19:GUS line (see The Plant Journal (2014), 77, 393-403) can be confirmed to respond to auxin. Using this promoter IAA19:GUS line, the inhibitory effect of compound 2 on auxin was confirmed.

1. Test method The Arabidopsis transformant promoter IAA19:GUS line (hereinafter referred to as the "transformant") described above is sown on a 1% sucrose-containing 1/2MS solid medium (Nippon Pharmaceutical Co., Ltd.) containing Compound 2 at a concentration of 40 μM. The plants were grown under long-day conditions (16 hours of light, 8 hours of darkness) at 23°C. As a comparative control, transformants were similarly inoculated and cultivated on a 1% sucrose-containing 1/2MS solid medium (Nippon Pharmaceutical Co., Ltd.) that did not contain Compound 2 (Cont).
10 days after seeding, the whole grown individuals were treated with GUS staining solution (2mM potassium ferricyanide, 10mM EDTA, 0.1% Triton X-100, 50mM Na Phosphate). ffer pH7.0, 0.5mM 5-bromo- Staining was performed with 4-chloro-3-indolyl β-D-glucuronide (X-Gluc) for 30 minutes at 37°C.
After staining, the reaction was stopped with ethanol:acetic acid=6:1 solution, decolorized with 100% ethanol solution, and observed with a stereomicroscope. This staining method specifically stained sites exhibiting auxin responsiveness.

2. Results The observed images are shown in Figure 27.
In the transformants grown in the medium not containing compound 2 (AAA1), as shown in the left image of Figure 27, many developing lateral root primordia were stained as sites showing auxin responsiveness (the ↓ area was stained). part).
On the other hand, in the transformants grown in the medium supplemented with Compound 2, the development of lateral root primordia stopped, and as shown in the right image of FIG. 27, there were no stained sites. In other words, Compound 2 was considered to reduce the auxin responsiveness of plants.
It has become clear that the anti-auxin action of the compounds of the present invention is exerted by reducing the auxin responsiveness of plants.

<Efficacy test of compound 2 on lateral root formation of naphthaleneacetic acid>
Naphthaleneacetic acid (also known as 1-naphthaleneacetic acid, hereinafter referred to as "NAA") is a plant growth regulator that exhibits auxin-like activity, and has effects such as regulating the number of fruits set, preventing fruit drop, promoting enlargement, and suppressing summer bud elongation. It is known to have. The effect of compound 2 on this NAA was tested.

1. Test method 1% sucrose-containing 1/2MS solid medium containing 50 nM and 100 nM NAA, and 1% sucrose-containing 1/2 MS containing 20 μM, 40 μM, and 100 μM compound 2 (AAA1) (Nippon Pharmaceutical Co., Ltd.) )) Prepare a solid medium, and add Arabidopsis wild type Col. Seeds of the -0 strain were sown and cultivated under long-day conditions (16 hours of light, 8 hours of darkness) at 23°C. The number of lateral roots was measured 10 days after sowing.

2. Test Results The measurement results of the number of lateral roots per 1 cm of taproot are shown in FIG.
At both NAA concentrations of 50 nM and 100 nM, the number of lateral roots per cm of the main root increased by the addition of NAA, but was suppressed in a concentration-dependent manner by the addition of Compound 2.

NAA is a different type of auxin than IAA. NAA is known as a type of auxin that is taken up into plants without an uptake carrier (see Planta (1996) 198: 532-541).
Compound 2 was confirmed to inhibit lateral root formation exhibited by NAA. This result confirmed that inhibition of the anti-auxin effect of Compound 2 does not inhibit auxin transport.

<Length suppression effect on broccoli>
A test was conducted to confirm the effect of Compound 2 on suppressing the growth of vegetable seedlings by densely planting them.
1. Test method A 288-hole cell tray was filled with seedling raising soil type B (Hokkaido Nozai Kogyo), and broccoli seeds (variety: Pixel (Sakata Seed)) were sown.
The plants were then cultivated in a greenhouse for 19 days, and then an aqueous solution of 0.1 mM, 1 mM, and 10 mM of Compound 2 to which 0.1% Approach BI (Kao Corporation) was added as a spreading agent was sprayed onto the leaves.
14 days after spraying, the maximum leaf length of the grown broccoli was measured. In addition, the investigation was conducted with 10 individuals and the number of repetitions was 2 (2 trays).

2. Test Results The results are shown in Figure 29. Furthermore, images of the measured seedlings are shown in FIG.
The maximum leaf length of the grown broccoli seedlings was reduced in size depending on the spray concentration of Compound 2. It was also confirmed that the spraying of Compound 2 shortened the plant height and resulted in sturdy seedlings.
In other words, it was revealed that Compound 2 suppresses the growth caused by dense broccoli cultivation. This was thought to result in suppression of the shade avoidance effect caused by auxin that occurs under dense planting conditions.

<Branch development effect test on pea>
1. Test method A 1/5000a Wagner pot was filled with Sukusuku Club 30 (Snow Brand Seedlings) as cultivation soil, and pea seeds (variety: Mijuka Kinukou (Snow Brand Seedlings)) were sown.
The plants were cultivated in a greenhouse for 50 days, and the growing point was removed at the 5th node.
Thereafter, an aqueous solution of 1 mM Compound 2 (AAA1) to which 0.1% Approach BI (Kao Corporation) was added as a spreading agent was sprayed onto the leaves. Foliar spraying was performed once a week.
For control (Cont), only the spreading agent-containing aqueous solution was sprayed onto the leaves in the same manner.
After that, cultivation was continued, and the number of branch nodes was counted four months after sowing.
The test group was repeated 2 times (2 pots), and the average value was calculated.

2. Results The results are shown in Figure 31. The number of branching nodes was 16 in the control, but when Compound 2 was sprayed, the number of branching nodes was 22.5, which clearly increased.
Compound 2 was found to have the effect of increasing the number of branches in pea.

<Spraying effect test on extending the flowering period of pot carnation flowers>
1. Test method: Potted carnations (variety: Shabon Rose (Snow Brand Seedlings)) at the flowering stage were irrigated with 100 ml of 1 mM AAA1 per plant.
The number of blooms and the number of wilted flowers were investigated from the day after application.
The number of repetitions was 3 (3 pots), the number of blooms and the number of wilted flowers were counted each day, and the average value was calculated.

2. Results Figure 32 shows the change over time in the number of blooms, and Figure 33 shows the change in the number of wilted flowers. On the last day of the observation period, the number of flowers in the pots treated with Compound 2 was 56.3, and the number of flowers in the untreated pots was 49.
Moreover, the cumulative number of wilted flowers did not appear during the period up to 15 days after the start of the test. After the 16th day, it increased over time. After the 17th day, the number of wilted flowers in the untreated control pot increased over time, whereas the increase in the number of wilted flowers in the pots treated with Compound 2 was significantly suppressed as shown in Figure 33. It had been. As shown in FIG. 34, the cumulative number of wilted flowers on the final day of observation was 6.7 on average in the pots treated with Compound 2, while it was 9.3 in the control.
On the final day of observation, the pots treated with Compound 2 remained in full bloom, while the pots without the additive (control) had fewer flowers and wilted flowers were noticeable. .
From the above test results, Compound 2 had the effect of suppressing the wilting of flowers after flowering, extending the flowering period, and enhancing the appearance of the flower pot.

<Length suppression effect on broccoli>
A test was conducted to confirm the effect of Compounds 9 and 10 on suppressing the growth of vegetable seedlings by densely planting them.
1. Test method A 200-hole cell tray was filled with seedling raising soil type B (Hokkaido Nozai Kogyo), and broccoli seeds (variety: Pixel (Sakata Seed)) were sown.
Then, they were cultivated in a constant temperature room (22°C) for 14 days, and then 1mM and 10mM aqueous solutions of compounds 9 and 10 were added to 0.1% Approach BI (Kao Corporation) as a spreading agent at a volume of 100ml ^{per 1m2} . Sprayed on leaves as shown.
Fifteen days after the spraying, the length of the true leaves of the grown broccoli was measured. In addition, the survey was conducted on 9 individuals and the number of repetitions was 3.

2. Test results The results are shown in Figure 40.
The length of the true leaves of the grown broccoli seedlings after spraying was reduced due to the spraying concentration of 10mM Compounds 9 and 10. Compound 9 tends to be smaller than compound 10 even at low concentrations.
That is, it was revealed that Compounds 9 and 10 suppressed the growth caused by dense broccoli cultivation. This was thought to result in suppression of the shade avoidance effect caused by auxin that occurs under dense planting conditions.

Claims (9)

An auxin and anthranilic acid activity inhibitor containing a compound represented by the following formula 1 or a salt thereof as an active ingredient.
(However, A is a single bond via a -CH(=O)-NH- or -CH=CH- structure, R1 is a hydrogen atom, a halogen atom, or an alkyl group, and R2 is a hydrogen atom, A halogen atom, an alkyl group having 1 to 6 carbon atoms, R3 is a hydrogen atom, an alkyl group, or a halogen atom, R4 is OH or OCH3, and R5 is an amino group, a halogen atom, a hydroxyl group, or hydrogen (R1 and R2 may form a closed benzene ring.) The activity of auxin and anthranilic acid according to claim 1, wherein the active ingredient is a compound, a salt or an ester thereof, in which the halogen atoms of R1, R2, and R3 are fluorine atoms, and the alkyl groups of R1, R2, and R4 are methyl groups. inhibitor. An auxin and anthranilic acid activity inhibitor containing as an active ingredient one or more compounds selected from the following compounds (1) to (15).
(1) 4-[(phenylamino)carbonyl]-benzoic acid (2) 2-amino-4-[(phenylamino)carbonyl]-benzoic acid (2- Amino-4-[(phenylamino)carbonyl]-benzoic acid)
(3) Methyl-2-Amino-4-[(phenylamino)carbonyl]-benzoate
(4) 2-Amino-4-[(1-naphthylamino)carbonyl]-benzoic acid
(5) 2-Amino-4-[(2,3-dimethylphenylamino)carbonyl]-benzoic acid
(6) 2-Amino-4-[(2-fluorophenylamino)carbonyl]-benzoic acid
(7) 2-Amino-4-[(3-fluorophenylamino)carbonyl]-benzoic acid
(8) 2-Amino-4-[(4-fluorophenylamino)carbonyl]-benzoic acid
(9) 4-(benzoylamino)benzoic acid
(10) 2-hydroxy-4-(benzoylamino)benzoic acid (11) 2-Chloro-4-(benzoylamino)benzoic acid (2-Chloro-4- (benzoylamino)benzoic acid)
(12) 4-(4-fluorobenzoylamino)benzoic acid
(13) 4-Stilbenecarboxylic acid
(14) 2-amino-4-(benzoylamino)benzoic acid
(15) 2-Amino-4-styrylbenzoic acid A plant growth regulator comprising the auxin and anthranilic acid activity inhibitor according to any one of claims 1 to 3. A lateral root development inhibitor comprising the auxin and anthranilic acid activity inhibitor according to any one of claims 1 to 3. A taproot elongation promoter comprising the auxin and anthranilic acid activity inhibitor according to any one of claims 1 to 3. A plant overgrowth inhibitor comprising the auxin and anthranilic acid activity inhibitor according to any one of claims 1 to 3. A branch development promoter comprising the auxin and anthranilic acid activity inhibitor according to any one of claims 1 to 3. A flower freshness-preserving agent comprising the auxin and anthranilic acid activity inhibitor according to any one of claims 1 to 3.