JPH02254A - Novel dipeptide compound - Google Patents

Abstract

NEW MATERIAL:A compound of the formula (R is H or protecting group for the amino group; Y is H or OH which may have a protecting group; X is an acidic amino acid residue which may have one or more protecting groups) and a salt thereof. USE:An agricultural and horticultural agent having a plant growth controlling action. Since the compound exhibits a remarkable action especially against a plant whose growth is depressed under the stress of low temperature, the compound is a ready and economical agricultural and horticultural agent. PREPARATION:The compound of the formula is synthesized by a conventional method in the peptide chemistry. A condensation method for forming the peptide bond includes an azide method, active ester method, mixed acid anhydrides method, acid chloride method, enzymatic condensation method and method using a condensing agent (e.g. N, N'-dicyclohexylcarbodimide).

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a novel dipeptide compound having plant growth regulating activity.

(Prior Art) In recent years, the food problem is predicted to become more serious, and efforts are needed to secure and even increase the production of agricultural products, and much research is being conducted. They believe that it is important to minimize the reduction in agricultural production due to abnormal weather, etc., and have been searching for compounds that can normalize the growth of agricultural crops exposed to low-temperature environments. As a result, the present inventors discovered that the dipeptide compound of the present invention has a plant growth regulating effect, and completed the present invention.

(Problems to be Solved by the Invention) An object of the present invention is to provide a novel dipeptide compound having a plant growth regulating effect and an agricultural and horticultural agent containing the compound as an active ingredient.

(Means for solving the problems) The dipeptide of the present invention is a novel compound represented by the following general formula (1).

R-HN -CHz -CHCHz -CO-X [wherein, R is hydrogen or a protecting group for an amino group, Y is hydrogen or a hydroxyl group which may have a protecting group, and X is one or more protecting groups. In the present invention, peptides and amino acids are designated by the International Union of Pure and Applied Chemistry (ItlP).
AC) - represented by abbreviations adopted by the International Union of Biochemistry (IUB) and abbreviations frequently used in the field; for example, the following abbreviations are used.

GABA: γ-aminobutyric acid GABOB: β-hydroxy-T-aminobutyric acid G
lu: glutamic acid Asp: aspartic acid In the above general formula (1), X is Glu%D -Glu, A
sp.

It represents an acidic amino acid residue such as D-Asp, and the amino acid residue may have one or more protecting groups. For example, preferred compounds include GABA Glu, GABA-D
-Glu.

GABA-As11. GABA-D-Asp,
GABOB-Glu, GABOB-D-Glu
, GABOB-Asp, GABOB-D
-Asp etc.

The dipeptide compound of the present invention can be produced by conventional methods in peptide chemistry. That is, the dipeptide compound of the present invention can be synthesized by either a liquid phase method or a solid phase method.

Condensation methods for forming peptide bonds include azide method, active ester method, mixed acid anhydride method, acid chloride method, enzymatic condensation method, N,N''-dicyclohexylcarbodiimide (DCC), and water-soluble carbodiimide. , N, N'
- a method using a condensing agent such as carbonyldiimidazole;
Alternatively, N-hydroxysuccinimide, N
Conventional peptide condensation methods can be used, such as the DCC-additive method using additives such as -hydroxy-5-norbornene-2,3-dicarboximide, 1-hydroxybenzotriazole, and the like.

Amino acids that serve as raw materials for the condensation reaction can be those that have commonly used protecting groups, and amino groups and side chain functional groups that do not participate in the reaction can be protected by known methods, or The carboxyl group or amino group may be activated.

As the protecting group for the amino group that does not participate in the reaction, the protecting groups used in normal peptide synthesis can be used.
Lower alkoxycarbonyl groups such as butoxycarbonyl and t-pentoxycarbonyl, aralkyloxycarbonyl groups such as benzyloxycarbonyl, or substituents such as O-chlorobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, and p-methoxybenzyloxycarbonyl. Examples include an aralkyloxycarbonyl group having an aralkyloxycarbonyl group.

As the protecting group for the carboxyl group of acidic amino acid residues such as glutamic acid and aspartic acid, the protecting group for the carboxyl group used in ordinary peptide synthesis, i.e., substitution of aralkyloxy groups such as benzyloxy, p-methoxybenzyloxy, etc. Aralkyloxy group having a group, t-
Lower alkoxy groups such as butoxy, ' t-pentoxy,
Cycloalkyloxy groups such as cyclopentyloxy and cyclohexyloxy, or 4-pyrisiloxy groups can be used.

Furthermore, as the protecting group for the hydroxyl group of β-hydroxy-T-aminobutyric acid, the protecting group for the hydroxyl group used in ordinary peptide synthesis can be used, that is, the 7-ralkyl group such as benzyl,
Examples include aralkyl groups having substituents such as bromobenzyl and chlorobenzyl, and lower alkyl groups such as t-butyl.

These substituents can be removed by conventional means such as catalytic reduction and acid decomposition.

The reaction temperature, time, solvent, etc. in the condensation reaction and deprotecting group reaction can be set according to reaction conditions used in ordinary peptide synthesis.

The dipeptide compound of the present invention includes pharmaceutically acceptable salts thereof, such as inorganic salts with alkali metals such as sodium and potassium, alkaline earth metals such as calcium and barium, and other metals such as aluminum; Examples include salts of ammonium and organic amines.

Further, the compound of the present invention includes its metal complex compounds, such as complex compounds with zinc, nickel, cobalt, copper, iron, etc.

These salts and metal complex compounds can be prepared by known methods.
They can be produced from the dipeptides of the present invention separately, or they can be converted into each other.

The amino acid residue in the present invention may be any one of a monomer, a monomer, and a monomer.

The obtained compound of the present invention is purified by conventional means such as chromatography and recrystallization, and subjected to elemental analysis, melting point, and NMR.
, IR, , LIV, mass spectrometry, etc. were used for identification. Note that the specific optical rotation was measured using the D-line of sodium.

An example of the manufacturing method of the present invention will be explained below using Examples.

In the following examples, the following abbreviations for substituents, reagents, etc. are used.

2: Benzyloxycarbonyl 0Bzl: Benzyloxy TosOH: Toluenesulfonic acid EDC, HC1: 1-Ethyl-3-(3'-dimethylaminopropyl)-carbodiimide hydrochloride (Example) Example 1゜+1) 5.36 g (D) ethylamine, 11.
9 g (7) Z-GABA-011 and 26.
5 g TosOH-Glu(OBzl)-0Bzl
Dissolved in 250-g of methylene chloride and cooled with water to give 53+ g
Nol EDC and 11Cl were added, and the mixture was stirred at 0°C for 2 hours and at room temperature for 20 hours.

After distilling off the solvent under reduced pressure, the residue was dissolved in ethyl acetate-water, and the organic layer was separated. After extracting the aqueous layer with ethyl acetate, the organic layers were combined, mixed with 10% citric acid aqueous solution, water, 5%
% sodium bicarbonate aqueous solution and saturated brine. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to obtain white crystals.

This was analyzed by silica gel column chromatography (eluted with ethyl acetate/chloroform-1/1), and 17.
4g of Z-GABA-Glu(OBzl)-08
I got zl.

Yield: 64% Melting point: 94-95°C [α]Ft': +1.0 (cL CHCl3) The following compound was obtained in the same manner.

Z -GABA -D -Glu(OBzl) -0B
zl Yield: 69% Melting point: 92-93℃ (α) ” Knee 1.1 (cl, CHCl5)Z-
GABA-Asp(OBzl) -08zl yield near 0
% Melting point: 105-106℃ (α) ”: +15.4 (cL CHCl
5) Z-GABA-D-Asp(OBzl)-08
zl Yield near 0% Melting point: 104-106℃ (α) ”: -15,9 (cl, CHCl
5) Z-ROL-GABOB-Glu(OBzl)-0
8zl white powder crystals + 2117.4g of Z-GABA-G 1u (OBzl
)-08zl, methanol 200wd, acetone 50-
and water lO−, and catalytic reduction was performed in the presence of 109 (palladium-carbon catalyst) at room temperature and under a hydrogen atmosphere at normal pressure. After 20 hours, the disappearance of the raw material was confirmed by thin layer chromatography. After that, the catalyst was removed by filtration.The filtrate was distilled off under reduced pressure, and water and toluene were added to the residual oil.
Acetic acid was azeotropically removed under reduced pressure. The residual oil was crystallized from ethanol and then recrystallized from water-ethanol to obtain 6.6 g of GIB^-Glu (compound 1) in the form of a white powder.

Yield: 89% Melting point: 17B-180℃ (d)'': -4,9 (cl, HtO) Elemental analysis: C*H+hNxOs 0.2H*O as
% N% N% Calculated value: 45.84 7.01 11.
8B actual value? 46.02 7.3G
11.75-position MR (0.1NDgO, internal standard t-Bu
O mouth δ-1,23) +1.91-2.02 (3tl,
m), 2.13-2.22 (11, ugly), 2.
42 (2H, t, J=7.5Hz), 2.46 (2H
, t, J-71Lz), 3.02 (2H, t, J-
7,5Hz), 4.32 (IH, dd, J-5,9H
z) The following compounds were obtained in the same manner.

GABA-D-Glu (Compound 2) Yield: 86% Melting point: 175-176°C (d) ”: +5.0 (cl+ Hg0) Elemental analysis: Cq H+ h N t 0%・0.2H1
0% N% N% Calculated value: 45.84 7.01 11°88 Actual value: 45.84 7.11 1
1.7ONMR (0,1NDzO, internal standard t-BuO
Mouth δ-1,23): 1.92-2.04 (3H,-)
, 2.15-2.24 (IH, m), 2.43
(211,t, J=7.5Hz), 2.48(2H,
t, J-711z), 3.02 (2H, t, J=7.
5Hz), 4.37 (111, dd, J-5,911
z) GABA-Asp (Compound 3) Yield: 92% Melting point: 184-185°C (d)”: +11.7 (cL Hid
) Elemental analysis: C* HI 4 N * 0% N% N% Calculated value as Os: 44.03 6.47 12.
84 actual value: 44.13 6.79 1
2.5ONMR (0,1NO!O, internal standard t-BuO
D δ−1,23) +1.95(2H,tt, J=7
．． 5. 7.5Hz), 2.43 (21,t.

J-7,5Hz), 2.93 (IH, dd, J=7.
17Hz), 2.98 (18, dd, J-5, 17H
2), 3.01 (211,t, J-7,5Hz).

4.77 (IH, dd, J-5,7Hz) GABA -
D-Asp (Compound 4) Yield: 89% Melting point: 180-182°C (d) ”: -11,0 (cl, HtO) Elemental analysis: 0% as C@HIaNzOs
N% N% Calculated value: 44.03 6.47 12.
84 actual measurement value: 43.98 6.67 1
2.62-position MR (0, INO!0. Internal standard t-BuO
D δ-1,23): 1.95 (21 positions, tt, J
-7,5,7,5112), 2.42(2H,t.

J-7, 5Hz), 2.87 (IH, dd, J-7,
17Hz), 2.94 (IH, dd, J-5, 171
1z), 3.10 (21+, t, J-7,5Hz).

4.69 (IH, dd, J-5, 7Hz) 0L-GAB
OB-Glu (Compound 5) Yield: 62% N? IR(0,1ND!0.Internal standard tlE t-BuO
Mouth δ, 1.23): 1.95-2.06 (IH, m)
, 2.17-2.26 (IH, m), 2.47
2.53 (211, a+), 2.54-2.58 (
2H, m), 2.96-3.03 (1B, m),
3.16-3.22 (IH, +w), 4.23-
4.31 (III, m).

4.40-4.46 (IH, a+) (Effect) 1. Plant growth regulating effect The plant growth regulating effect of the compound of the present invention was investigated using rice (Nipponbare) stored at 4°C.

A filter paper soaked with the test water collection solution (I XIO-6M) was placed in a Petri dish to serve as a germination bed, and test seeds were sown.

The seeds that germinated on the fourth day were transferred to a plant culture test tube that did not contain the test aqueous solution, and a growth test was conducted. Seven days after transplantation, the above-ground and underground parts were weighed.

- The test was conducted in a dark place at 20°C with 30 individuals in the group, and the average value and standard error were determined.

Examples of the results are shown in Tables 1 and 2.

(Umbrella Umbrella Umbrella: p < 0.0011 Rate: p < o, o
s, *: p < 0.01) Test drug control Compound 1 Compound 2 Compound 3 Compound 4 Test drug control Compound 1 Compound 2 Compound 3 Compound 4 1st Upper surface part (ms) 44.9 55.8 55. 5 57.1 54.8 0.98 1.02”” 1.08”” 0.89s0 0.91°0 2nd outer ground upper type (Akebono) 21.4 26.1 26.1 25 .2 24.5 0.58 0.71°11 0.98” 0.77” 0.66” Seed root (am) 49.2 61.5 55.2 63.3 66.8 2.70 2. 83” 4.67 3.32” 3.3411 Underground type (qr) 23.6 30.8 30.0 24.6 28.3 1.89 1.23” 0.74” 1°28 1. 86 (Effect) As is clear from the above results, the compound of the present invention has an excellent plant growth promoting effect.
Under low temperature stress (20℃, the optimum growth temperature for rice is approximately 30℃)
) has a particularly pronounced effect on plants whose growth has been inhibited,
It has the characteristic of normalizing plant growth.

In addition, this plant growth normalizing effect can be obtained by treating seeds with the compound of the present invention only during the germination period, so it is very simple and economical, and the compound of the present invention is extremely useful as an agricultural and horticultural agent. .

Claims (2)

[Claims] (1) General formula (I): ▲Mathematical formulas, chemical formulas, tables, etc.▼(I) [In the formula, R is hydrogen or a protecting group for an amino group, Y is hydrogen or a hydroxyl group that may have a protecting group, X represents an acidic amino acid residue which may have one or more protecting groups. ] and its pharmaceutically acceptable salts. (2) General formula (I): ▲Mathematical formulas, chemical formulas, tables, etc.▼(I) [In the formula, R is hydrogen or a protecting group for an amino group, Y is hydrogen or a hydroxyl group that may have a protecting group, X represents an acidic amino acid residue which may have one or more protecting groups. ] or a pharmaceutically acceptable salt thereof as an active ingredient.