JPH045290A - New adenine derivative - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to novel adenine derivatives. The adenine derivative according to the present invention exhibits cytokinin activity, promotes cell division in plants, promotes growth of lateral buds, promotes germination, promotes flower bud formation and flowering, promotes fruit set, enlarges fruits, suppresses aging, and promotes substance accumulation in storage organs. It is a substance useful for promoting the activity of plant physiological functions such as.

[Prior art, problems to be solved by the invention] Conventionally,
Numerous compounds have been found to regulate the growth of plants, especially grains, fruits, and vegetables. Among these, substances belonging to the group of substances collectively called cytokinins are known to have many physiological effects, and representative compounds include zeatin, kinetin, and benzyladenine. These cytokinins promote cell division in plants,
It is known to have physiological effects on plants such as promoting the growth of lateral buds, promoting germination, promoting flower bud formation and flowering, promoting fruit set, fruit enlargement, suppressing aging, and promoting substance accumulation in storage organs.

Substances exhibiting cytokinin activity have conventionally existed in nature, but the amount of these natural products is limited and they are not easily available. Furthermore, although substances exhibiting cytokinin activity have many physiological effects, their practical applications are currently limited.

One of the reasons for this limited practical use is that conventionally known cytokinins have poor solubility in water, and are not sufficiently absorbed into plants or translocated to other organs. It will be done.

The object of the present invention is to provide a synthetic substance exhibiting cytokinin activity that is more water-soluble than conventionally synthesized cytokinin and more easily available than natural cytokinin.

[Means for Solving the Problems] In view of the above-mentioned circumstances, the present inventors have conducted numerous studies and studies on compounds that exhibit cytokinin activity and are water-soluble. The present invention was achieved by discovering that novel adenine derivatives having a side chain as shown in the following formula A and their salts are effective.

CH3/(CH2), -N [:A)\CH3 That is, the present invention relates to adenine derivatives and salts thereof represented by the following general formulas ■ and general formulas (provided that in -Shinzo I and general formula ■) , n is an integer of 2 or 3,
X represents a hydrogen atom or a chlorine atom, and Y represents a hydrogen atom or a 2-ribosyl group. However, this excludes the case where both X and Y are hydrogen atoms.

In the ship's warehouse, 2 contains equivalent amounts of hydrochloric acid, hydrobromic acid, sulfuric acid,
Nitric acid, phosphoric acid, formic acid or acetic acid. ) The adenine derivative represented by the above general formula (2) according to the present invention can be obtained, for example, by the following method.

2.6-dichloropurine or 6-chloropurine riboside and an amine derivative having a structure represented by the following formula B are mixed with a trialkylamine (for example, triethylamine or ethyldiisopropylamine) in an organic solvent such as an alcohol. ) can be synthesized by heating and reacting in the presence of

OCH.

/ NH2(CH2)l, N (:B)
＼ H3 (However, n is an integer of 2 or 3) - Onboard In order to obtain the adenine derivative of the present invention represented by ■ in high purity, it is first necessary to obtain the amine derivative represented by formula B in high purity. It's important. A method for obtaining the amine derivative represented by formula B with high purity is, for example, by reacting N,0-dimethylhydroxylamine and N-(haloalkyl)phthalimide to obtain a phthalimide derivative of the amine represented by formula B, There is a way to hydrolyze this. In this case, 2,6
- In the reaction with dichloropurine or 6-chloropurine riboside, it is also possible to react with 26-dichloropurine or 6-chloropurine riboside using a hydrolysis reaction solution without isolating the amine of formula B. .

A typical example of the adenine derivative represented by the general formula I according to the present invention is 2-chloro-N6-[2-(N-
Methoxy-N-methylamino)ethylcoadenine, formula ■
2-chloro-N'-[3-(N-methoxy-
N-methylamino)propyl]adenine, N6-[2-(N-methoxy-N-methylamino) represented by the formula ■
Ethylcoadenine riboside is mentioned.

In addition, the adenine derivative according to the present invention can be easily obtained by a conventional method using mineral acid salts such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, or organic acids such as formic acid and acetic acid. It can be in the form of an acid salt.

(However, n is an integer of 2 or 3, X is a hydrogen or chlorine atom, and Y is a hydrogen atom or a 2-ribosyl group. However, this excludes cases where both X and Y are hydrogen atoms. Hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, formic acid or acetic acid.) Specifically, for example, 2-chloro-N'-[2-(N-methoxy-N- [methylamino)ethyl] adenine hydrochloride, an adenine derivative represented by the general formula (■) in the present invention, and -
Table 1 shows the physical properties of the adenine derivative salt represented by Shipura (■).

2 IV 3 CIH, -230 to 231 (
Decomposition) 3 V 2 Hribs -127
~1294 VI 2 [1'lHHCl18
2-184 (Degradation) The adenine derivative represented by the above-mentioned ship stock (■) and the salt of the adenine derivative represented by the general formula (■) according to the present invention have plant hormone and cytokinin activity, and can be used as plant growth regulators. It is expected that it can be used in fields or as a medium component for plant tissue culture.

Next, Examples and Test Examples will be shown regarding the synthesis of the adenine derivative according to the present invention and its physiological activity.

[Examples] The present invention will be explained in more detail with reference to Examples, but the present invention is not limited to these Examples.

Example 1 Synthesis of 2-chloro-N6-[2-(N-methoxy-N-methylamino)ethylcoadenine (compound 1) As follows, a side chain diamine was first synthesized, and this
- reacted with dichloropurine.

(1) N-[2-(N-methoxyl N-methylamino)
Synthesis of N,0-dimethylhydroxylamine hydrochloride 390g
Isopropanol suspension containing (40,0 mmol) 5
5.56 ml of triethylamine was added dropwise to 0 ml. The mixture became homogeneous after 5 minutes. 2.45g for this
(10.0 mmol) of N-(2-bromoethyl)phthalimide was added, and the mixture was heated under reflux for 29 hours.

After cooling the reaction mixture to room temperature, it was poured into 200 ml of saturated sodium bicarbonate solution, extracted with chloroporm, and the organic layer was washed with water and dried over magnesium sulfate. Chloroform was distilled off, and the residue was separated and purified by silica gel column chromatography. 1. By development with 30% hexane-chloroform.
37g of N-(2-chloroethyl)phthalimide was eluted, and when further developed with chloroform, 0.96g (yield 41%) of N-[2-(N-methoxy-N-methylamino)ethylcottalimide was white. Obtained as crystals.

Melting point = 130-132°C 'H-NMR (CDCl2.TMS internal standard); δ = 2
．． 40 (s, 3N), 2.93 (t, J=
6Hz, 2H).

3.53 (s, 3H), 3.93 (t, J=6Hz,
2H).

7,77 (m, 4H) ppm (2) Synthesis of N-methoxy-N-methylethylenediamine Imide obtained in the above (1) 1. 78g (7゜42
mmol) was dissolved in 3Qml of methanol, and 0.
408 g (8.16 mmol) of hydrazine hydrate was added, and the mixture was heated under reflux for 6 hours.

The reaction mixture was cooled in a water bath and the resulting solid was filtered off. The filtrate was concentrated to 4 ml using an evaporator in a 5° C. water bath, 10 ml of ether was added thereto, and the precipitated solid was filtered off. After concentrating the filtrate, distill it under normal pressure to give 0.324g.
(yield 42%, boiling point 97-101°C) of N-methoxy-
N-methylethylenediamine was obtained.

' H-NMR (CDCl2. TMS internal standard)
;δ=1.83(s, 2H), 2.60(s, 3
H), 2.78 (m, 4tl).

3,52 (s, 3H) ppm (3) 2-chloro-N6-[2-(N-methoxy-N
-Methylamino)ethyl] Adenine Synthesis 2.6-Dichloropurine 0. 189g (1,0Qmmol) and ethyldiisopropylamine 0°174ml (1,00
mmo 1) was dissolved in 3 ml of n-butanol, 0.104 g (1° OQ mmol) of N-methoxy-N-methylethylenediamine was added thereto, and the mixture was heated under reflux on an oil bath for 5 hours.

The reaction mixture was poured into 20 ml of water, extracted with methylene chloride, and the organic layer was dried over magnesium sulfate. The solvent was distilled off under reduced pressure, and the resulting solid was decolorized with activated carbon and then recrystallized from methanol-chloroform to obtain 0.11 g (yield: 43%) of the title compound as colorless columnar crystals.

Melting point: 232-233°C (decomposition) 'H-NMR (DMSO-dll/CD, 0D=2/1
．． TMS internal standard); δ=2.58 (s, 3H)
, 2.86 (t, J=6Hz, 2H).

3,51 (S, 3H), 3.73 (tm,
J=6Hz, 2H)8,00(s, IH)I]J]
ln Infrared absorption spectrum (KBr); νmax = 2
950", 1590", 1295', 1250'
.

1040", 935'car' Ultraviolet absorption spectrum; λmax (H2O) = 212 (20, 100),
270 (15,700) nmλmax (0,1HMCI
) =2I3'h(20,500), 274(14,
300) runλmax (0, lNNa[)H) =
276 (16,100), 286” (2,10
0) nm elemental analysis; Calculated value as C9Hl3CIN60 C: 42.11%) l: 5.10%
N: 32.74% Actual value C: 42.30% H: 4.98% [33,02
% Example 2 Synthesis of 2-chloro-N'-[3-(N-methoxy-N-methylamino)propyl]adenine (compound 2) First, a side chain diamine was synthesized as follows, and this was converted into 2, 6
- reacted with dichloropurine.

(1) N-[3-(N-methoxy-N-methylamino)
Synthesis of Provircostalimide 5.56 ml of isopropatol suspension containing 390 g (40.0 mmol) of N,○-dimethylhydroxylamine hydrochloride was added
of triethylamine was added dropwise. The mixture became homogeneous after 5 minutes. Add to this 2.68g (10,0mrnol
) of N-(3-bromopropyl)phthalimide,
The mixture was heated under reflux for 33 hours.

After cooling the reaction mixture to room temperature, it was poured into 200 ml of saturated sodium bicarbonate solution, extracted with chloroform, and the organic layer was washed with water and dried over magnesium sulfate. Chloroform was distilled off, and the residue was separated and purified by silica gel column chromatography. 1. By developing with 30% hethane-chloroform.
03g of N-(3-chloropropyl)phthalimide was eluted, and when further developed with chloroform, 1.21g (
N-[3-(N-methoxy-N-methylamine)probylcotta imide was obtained as white crystals in a yield of 53%.

Melting point: 55-57°C 'H-NMR (CDCl2.TMS internal standard); 64,
93<quin, J=6Hz, 2H), 2.55
(s, 3H).

2,68 (t, J=6Hz, 2H), 3.5
2 (s, 3H).

3.80 (t, J=6Hz, 2H), 7.75 (m,
4H) ppm (2) 2-chloro-N6- [3-(N-
Synthesis of methoxy-N-methylamine)propyl]adenine 0.540g (2.18mmol) of N-[3-(
Dissolve N-methoxy-N-methylamino)propylcottalimide in 20ml of methanol, add 0.12ml of
7 ml (2.62 mmol) of hydrazine hydrate was added, and the mixture was heated under reflux for 6 hours.

After the reaction mixture was cooled to room temperature, 30 ml of ether was added thereto, and the mixture was left at 0° C. for 1 hour to sufficiently precipitate a solid.

Solids were removed by suction filtration, and the filtrate was dried over sodium sulfate and concentrated to 2 ml using an evaporator.

This concentrated solution was dissolved in 5 ml of impropatul, and 0.095 g of 2,6-dichloropurine (0,503 ml
mo 1) and ethyldiisopropylamine 0.088m
1 was added thereto, and the mixture was heated under reflux on an oil bath for 5 hours.

The precipitated crystals were filtered off and then recrystallized from methanol to obtain 0.049 g (yield: 8%) of the title compound as a white powder.

Melting point = 230-231°C (decomposition) 'HNMR (DMSO-ds, TMS internal standard); δ = 1.33 (quin, J = 6Hz, 2H)
, 2.50 (S, 3H).

2,65 (t, J=6Hz, 2H), 3.4
4 (s, 3N).

3,60(m, 2H), 8.08(s, IH)p
pm infrared absorption spectrum (KBr); vmax
=2930v', 2780'', 1585', 15
40', 1420'.

1335''', 1240''', 1125', 10
30', 910'cm' Ultraviolet absorption spectrum; λmax (H2O) 212 (24,700
), 271 (18,700) nmλmax(0
, INHC'l) 212 (23,000), 272
(16,400)nmλmax (0,lNNaOH)
277 (16,900), 283sh (13
,900) nm elemental analysis: C+otLsCINsO
Calculated value as C: 44.37% H: 5.58%
N: 31.04% Actual value C: 44.51% )l: 5.53% N: 30.
82% Example 3 Synthesis of N6-[2-(N-methoxy-N-methylamino)ethyl]adenine riboside (compound 3) 6-chloropurine riboside 0.110 g (0°384 m
mo 1), N-methoxy-N-methylethylenediamine 0.130 g (1,21 mmol) and ethyldiisopropylamine 0.174 ml (1,00 mmol 1)
was dissolved in 5 ml of n-butanol and heated under reflux on an oil bath for 5 hours.

The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (developing agent: ethanol/chloroform = 1/
4) to obtain 0.034 g (yield 25%) of the title compound as a white solid.

Melting point = 127-129°C 'H-NMR (CD, 00/CDC13 = 1/3.TM
S inside 111); δ=2.63 (s, 3H),
2.90 (t, J=6Hz, 2H), 3.
54 (s, 3H).

3,80 (m, 4H), 4.1-4.4 (m,
2H), 4.91 (t, J=6Hz, IH).

5, 85 (d, J=6Hz, IH), 8.00 (
s, LH), 8.20 (s, LH) ppm infrared absorption spectrum (KBr); vmax = 3100
", 2910', 1605', 1585', 1
330”.

1285'', 1085'', 1040', 755°
cm ultraviolet absorption spectrum; λmax (H2O) = 210 (20,400),
266 (21,000) nmλmax (0,1NHC
I) =266 (19,100) nmλmax (
0,1NNaOH) =266 (21,400) n
Calculated value as m elemental analysis + Cl4H2□~s[]s C: 47.45% H: 6.26% N: 23.
72% actual value C: 47.58% H: 6.25% N+23.4
5% Example 4 Preparation of 2-chloro-N6-[2-(N~methoxy-N-methylamine)ethyl]adenine hydrochloride (compound 4) 0.270 g (1,05 mm○1) of 2-chloro- N
Dissolve 6-[2-(N-methoxy-N-methylamino)ethyl]adenine in 15 ml of ethanol and add 1. 05 ml of 1N HCl was added.

The solvent was distilled off under reduced pressure, and the residue was washed with chloroform.
．． 290 g of crude product was obtained. This was recrystallized from ethanol to obtain 0.275 g (yield: 8'9%) of the title compound as colorless columnar crystals.

Melting point: 182-184°C (decomposition) 'H-NMR (DMSO-d6. TMS internal standard); δ = 2.72 (S, 3H), 3.07
(t, J=5Hz, 2N).

3,61 (s, 3H), 3.73 (m, 2
H), 8.60 (br, LH).

8,71 (s, IH) ppm Infrared absorption spectrum (KBr); νmax = 2
920', 2500br, 2150'', 1600
', 1550''.

1235', 1140'', 925°cm-' Elemental analysis; Calculated value as C, H, 4C12N6 [1 C: 3
6.87% H: 4.81% N: 28.67% Actual value C: 36.99% H: 4.83% N: 28.3
6% Test Example 1 Cytokinin activity test based on chlorophyll retention effect Rice seeds (variety: Southeast No. 11) were sown in a seedling box filled with soil and kept in a glass room (25°C during the day/15°C at night) for about 1 month. I let it grow.

A leaf piece with a length of 1 cm was cut from the center of the fourth leaf when the sixth leaf was expanded. 2 ml of test solution containing the specified concentration of test compound
A set of five cut leaf pieces were floated in a glass tube with an inner diameter of 32 mm. After being left at 30°C in the dark for 3 days, the leaf pieces were placed in a test tube containing 10ml of 80% ethanol and immersed in a hot bath at 80°C for 20 minutes to extract chlorophyll.

After cooling, add 80% ethanol to make 10ml.
Absorbance was measured at a wavelength of 65 nm.

The aging suppression rate due to the chlorophyll retention effect in the treated plot compared to the untreated plot was calculated using the following formula.

The results are shown in Table-2.

Concentration (mg/l) Test Example 2 Cytokinin Activity Test by Promoting the Synthesis of Betacyanin in Pygmy Corylus A plastic container (19 x 28 cm) was lined with two layers of filter paper and moistened with 60 ml of distilled water.

Seeds of Himogeido were sown on filter paper and kept in the dark at 27°C for 3 days to germinate. Shoots of uniform size were selected, the upper part of the hypocotyl was cut, and the seed coat was removed.

This cotyledon and the upper 3 to 4 mm of the hypocotyl were used for the assay. A glass tube with an inner diameter of 32 mm was lined with filter paper, and the test compound at a predetermined concentration and 0.0 g/l containing 0.5 g/l of tyrosine were placed in a glass tube with an inner diameter of 32 mm.
．． 0065M potassium phosphate buffer (pH 6,3)
Add 1 m, arrange 10 cotyledon sections, and place in the dark at 27
It was kept at ℃ for 20 hours. Ten cotyledon sections were placed in a test tube containing 3 ml of distilled water, frozen and thawed three times to extract betacyanin, the absorbance of the solution was measured, and the amount of pigment was determined from the difference in absorbance at wavelengths of 542 nm and 620 nm. Quantitated.

The results are shown in Table-3.

Test Example 3 Test on absorption and mobility in rice; Penetration and chlorophyll retention effect of Compound 1 and benzyladenine on rice leaf pieces Rice seeds (variety: Southeast No. 11) were sown in seedling boxes filled with potting soil, and placed in a glass room (during the day). Approximately 1 at 25℃/15℃ at night)
It was grown for a month.

A leaf piece with a length of 33 mm was cut from the center of the 5th leaf when the 7th leaf was expanded. A piece of circular filter paper was placed in a Petri dish with a diameter of 9 cm and moistened with 2.5 ml of distilled water. A slide glass was placed on top of this, and five cut leaf pieces were arranged on top of this slide glass so that the surface of the leaf pieces was facing upward. 0.0% of the test compound was placed in the center of this leaf. D2% Sorbo
10 μl of a test solution prepared by dissolving Ru 8214 (manufactured by Toho Chemical Industry Co., Ltd.) in an aqueous solution to a concentration of 10 mg/l was placed on the plate. After the petri dish was covered, it was placed in the dark at 25° C. for 5 days, and the length of the green part remaining due to the chlorophyll retention effect of the test solution was measured, centering on the area where the test solution was placed.

In addition, for the untreated plot, 10 μm of 002% Tsurupol-8214 (manufactured by Toho Chemical Industry Co., Ltd.) was placed on the center of the leaf. The test was performed in two replicates.

The results are shown in Table 4. In comparison at the same concentration, Compound 1 had a larger green area than benzyladenine, indicating better absorption and mobility.

Reference Example The solubility in water of the compound of the present invention and a conventionally known cytokinin substance was measured. As shown in the following table, the results showed superior solubility compared to conventionally well-known representative substances.

[Effects of the Invention] The adenine derivative according to the present invention not only exhibits cytokinin action, but also has higher water solubility and better water solubility than conventional synthetic products.
It has excellent permeability into living organisms and is expected to be used as a plant growth regulator.

benzyladenine Zeatin kinetin 0, 24 0, 18 14, 2 1.3 X1O-3 0, 14 X10-3

Claims (1)

[Claims] An adenine derivative represented by the following general formula I and a salt of an adenine derivative represented by the general formula II. ▲There are mathematical formulas, chemical formulas, tables, etc.▼[I] ▲There are mathematical formulas, chemical formulas, tables, etc.▼[II] (However, in general formula I and general formula II, n is 2
or an integer of 3, X represents a hydrogen atom or a chlorine atom, and Y represents a hydrogen atom or a 2-ribosyl group. However, this excludes the case where both X and Y are hydrogen atoms. In general formula II, Z is an equivalent amount of hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, formic acid or acetic acid. )