WO1996003371A1 - N-acyl-n-(substituted cinnamoyl)ethylenediamine derivative - Google Patents

Abstract

A novel N-acyl-N-(substituted cinnamoyl)ethylenediamine derivative represented by general formula (I) and serving as an antiphlogistic and/or NGF production accelerator, (wherein R1 represents the acyl residue of a C¿16? or higher unsaturated fatty acid; R?2 and R3¿ represent each independently hydrogen or lower alkyl; and n represents 1 or 2). This compound is expected to be usable as, for example, an antiphlogistic because it has the activities of O¿2?- production inhibitor in human neutrophilic leukocytes and intracellular phospholipase A¿2? inhibition, and as a remedy for dementia, spinal cord injury, peripheral nerve injury, diabetic neuropathy and amyotrophic lateral sclerosis because it has the activity of accelerating NGF production.

Description

 Specification

N-acyl-N-substituted cinnamoylethylenediamine derivatives

The present invention, 0 ₂ _ production inhibitory activity, intracellular Hosuhoripa Ichize A ₂ inhibitory activity, and / or N with nerve growth factor (NGF) production promoting activity - § Shiru N- substituted cinnamoyl ethylenedioxy § Mi emissions It concerns derivatives. Background art

The inflammatory response is a kind of host defense effect that is activated when a harmful stimulus invades the living body. However, as a result, swelling, pain, organ dysfunction, and other troubles are caused, and death often occurs. From acute inflammation associated with type 1 allergy to external effects to chronic inflammation due to nephritis and rheumatic diseases, the causes, developmental processes, and symptoms are extremely wide and complex. A drug called an anti-inflammatory agent is used as this symptomatic treatment, and is roughly classified into steroidal anti-inflammatory drugs and non-steroidal anti-inflammatory drugs, and various glucocorticoid-indomethacin etc. are representative. . However, steroid anti-inflammatory drugs have an inhibitory effect on protein synthesis including various proteinaceous mediators, and have a wide range of pharmacological effects and a large curative effect, but are known to cause serious side effects. It has recently been shown that the separation of pharmacological and side effects is almost impossible. On the other hand, non-steroid anti-inflammatory drugs mainly act as cyclooxygenase inhibitors (suppress prostaglandin production). The effect is limited due to its pharmacological action. Therefore and development of anti-inflammatory agent is required based on a new pharmacological action, 0 ₂ - production inhibitory activity, the compounds having intracellular phospholipase A ₂ inhibitory activity is considered a substance that meets the purpose.

That is, the force various active oxygen species are the most central mediators involved in tissue damage during inflammation, ', its production leukocytes (neutrophils, macrophages) due to 0 ₂ _ production by. It is known that this active oxygen is involved in various diseases such as duodenal ulcer, gastric sac, arteriosclerosis, ischemic disease of brain and heart, cancer, aging, cataract, autoimmune disease, inflammation, arthritis, and edema. Therefore, the potential of this inhibitor as a medicine has been studied [Pharmacia, Vol. 29, No. 9, 1014, 1029 (1993)].

Also an enzyme recently its presence intracellular Hosuhorino instrument over peptidase A ₂ is Akiraraka, the main inducing inflammation Meje one coater der Ru prostaglandins by its activation, leuco Toryen, platelet activation It is known that the enzymatic production of factor (PAF) is commonly initiated [proteins, nucleic acids, enzymes, Vol. 36, No. 3, 325 (1991)]. PAF is a powerful inflammatory mediator along with prostaglandins and leukotrienes, but its synthesis inhibitor is not yet known. Thus, intracellular phosphorylase Horipaze eight ₂ is considered useful as a prophylactic or therapeutic agent for inflammatory disorders and allergic diseases.

On the other hand, NGF differentiates nerve cells in vitro to promote neurite outgrowth, maintains the survival of nerve cells, and administers NGF into the brain in animal experiments to improve the memory-learning effects of old rats. It is known to be improved. It is also known to prevent neuronal cell death from cerebral ischemia: J. Neurosci., 6, 2155 (1986), Brain Res., 293, 305 (1985), Science 235, 214 (1986λ Proc. Natl. Acad. Sci. USA, 83, 9231 (1986), Annals of Neurology, 120, 275 (1986, Chemistry and Biology, Vol. 29 , No. 10, 640 (1991)].

 In Alzheimer's senile dementia, it has been confirmed in many cases that most of the cholinergic neurons of the Meinert nucleus, which are the nerve cells that control memory and thinking, are killed and lost. It has been clarified that NGF is essential for survival and differentiation [Geriatric Neurology, 3, 751 (1996)]. Lance and Olson et al. Reported that mouse NGF (6.6π ^ Ζ3 months) was administered intraventricularly to Alzheimer's disease patients to improve the symptoms of dementia (1991 Alzheimer's Disease Treatment Symposium). .

 In addition, NGF acts as a trophic factor not only in the central nervous system but also in peripheral perception and the sympathetic nervous system, and is an essential factor for nerve regeneration. That is, it is considered that it can be used for the treatment of spinal cord injury, peripheral nerve injury, diabetic neuropathy, amyotrophic lateral sclerosis and the like.

 However, in order to actually use NGF for treatment of humans, mass production of NGF has not been established, and NGF does not pass through the blood-brain barrier, which makes it inconvenient to administer NGF from the periphery. There is. Disclosure of the invention

The present inventors have results of various search specific Ν- Ashiru Ν- substituted cinnamoyl ethylenedioxy § Mi emissions derivatives, 0 ₂ - production inhibitory activity, the intracellular host Suhoripaze A ₂ inhibitory activity and Ζ or NG F production The inventors have found that they have a promoting activity and completed the present invention.

That is, the present invention provides the following general formula _{R 'NH- CH 2 CH E NH} irC -CH = in CH represented by N- Ashiru -N- substituted cinnamoyl ethylenedioxy § Mi emissions derivative conductor (wherein, R ¹ is higher unsaturated one 6 or more carbon atoms Represents an acyl residue of a fatty acid, R ² and R ³ independently represent a hydrogen atom or a lower alkyl group, and n is 1 or 2), and 0 ₂ -production inhibitory activity containing the active ingredient as an active ingredient; It provides intracellular phospholipase ₂ inhibitory activity and NGF production promotion activity.

 Examples of the acyl residue of the higher unsaturated fatty acid having 16 or more carbon atoms in the above general formula include a hexadecenoyl group, an oleoyl group, a linoleoyl group, a linolenoyl group, an eicosatrienoyl group, an arachidonyl group, and an eicosapentaenoyl group. And a docosatetraenoyl group and a docosahexaenoyl group. The lower alkyl group means an alkyl group having 1 to 6 carbon atoms, and examples thereof include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a pentyl group, and a hexyl group.

The N-acyl-N-substituted cinnamoylethylenediamine derivative of the present invention can be produced, for example, according to the following scheme.

'OH o ϋ r H ₂ N-eCH _z CH ₂ NH rH

 Condensing agent,

R 'NH- CH ₂ CH _z NH rH

HO— C

本発明の各薬剤は、 治療のために経口的あるいは非経口的に投与 することができる。 経口投与剂としては散剤、 顆粒剤、 カプセル剤、 錠剤などの固形製剤あるいはシロップ剤、 エリキシル剤などの液状 製剤とすることができる。 また、 非経口投与剤として注射剤とする ことができる。

Each agent of the present invention can be administered orally or parenterally for treatment. For oral administration, solid preparations such as powders, granules, capsules and tablets or liquid preparations such as syrups and elixirs can be used. In addition, it can be prepared as an injection for parenteral administration.

 These preparations are produced in a conventional manner by adding a pharmacologically and pharmaceutically acceptable production aid to the active ingredient. Furthermore, it is also possible to prepare a sustained-release preparation by a known technique. When the production aid is used, the amount of the N-acyl-N-substituted cinnamoyl ethylenediamine derivative in each drug of the present invention is usually 0.1 to 10% by weight, preferably 0.2 to 5% by weight. %.

To prepare a solid preparation for oral administration, the active ingredient must be mixed with excipients such as lactose, starch, crystalline cellulose, calcium lactate, magnesium metasilicate, magnesium anhydride, etc., to give a powder. If necessary, add a binder such as sucrose, hydroxypropylcellulose and polyvinylpyrrolidone, and a disintegrating agent such as carboxymethylcellulose and calcium carboxymethylcellulose. Alternatively, dry granulation is performed to obtain granules. To produce tablets, these powders and granules may be compressed as they are or by adding a lubricant such as magnesium stearate or talc. These granules or tablets are coated with an enteric base such as hydroxypropylmethylcellulose phthalate, methacrylic acid, methyl methacrylate copolymer, etc., and then coated with enteric-coated preparations, or coated with ethyl cellulose, carnauba wax, hydrogenated oil, etc. Formulation. To prepare capsules, powders or granules are filled into hard capsules, or the active ingredient is dissolved as it is or dissolved in glycerin, polyethylene glycol, sesame oil, olive oil, etc., and then coated with a gelatin film to prepare soft capsules. can do.

 To manufacture a liquid preparation for oral administration, the active ingredient and a sweetener such as sucrose, sorbitol, and glycerin are dissolved in water to give a clear syrup, and then an elixir by adding essential oils and ethanol. Emulsion or suspension may be prepared by adding arabia, gum tragacanth, polysorbate 80, carboxymethylcellulose sodium and the like. If desired, flavoring agents, coloring agents, preservatives and the like may be added to these liquid preparations.

In order to manufacture an injection, the active ingredient is adjusted to a pH adjusting agent such as hydrochloric acid, sodium hydroxide, lactose, sodium lactate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride, Dissolve in distilled water for injection together with a tonicity agent such as glucose, filter aseptically and fill the sample, or freeze dry in a vacuum after adding mannitol, dextrin, cyclodextrin, gelatin, etc. It may be a dissolvable injection agent. The active ingredients are lecithin and polysorbate 8 0. Polyoxyethylene hydrogenated castor oil and the like can be added and emulsified in water to prepare an emulsion for injection.

 In order to manufacture a rectal preparation, the active ingredient and a suppository base such as cocoa butter, a fatty acid tri-, di- or monoglyceride, or polyethylene glycol are moistened, melted, poured into a mold and cooled, or The active ingredient may be dissolved in polyethylene glycol, soybean oil, or the like, and then covered with a gelatin film.

 Each drug of the present invention having the above constitution can be produced by a known production method, for example, a method described in the Japanese Pharmacopoeia 10th Edition General Rules for Preparations or a method with appropriate improvements. BEST MODE FOR CARRYING OUT THE INVENTION

 Next, the present invention will be described specifically with reference to examples, but the present invention is not limited to these examples. Reference Example 1. Synthesis of N-4, 7.10, 13, 16, 19-docosahexaenoylethylenediamine (MA-EDA)

3. 28 g (10 mmol) of docosahexaenoic acid (95% or more) and 1.78 g (II bandol) of N, N-carbonyldiimidazole are dissolved in anhydrous tetrahydrofuran (hereinafter abbreviated as THF) I ODIU, The reaction was carried out at room temperature for about 1 hour under a nitrogen stream. Then, a solution of 1.5 g (25 mmol) of ethylenediamine (hereinafter abbreviated as EDA) and 2.55 g (25 marl ol) of triethylamine dissolved in 10 mL of anhydrous THF was added to the reaction mixture under ice-cooling. The reaction was performed for 2 hours. After completion of the reaction, 60 mL of 0.1N hydrochloric acid and 300 mL of chloroform-methanol (2: l) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. A small amount of Silica gel dissolved in form and pre-activated with chloroform

(Place on a 160 gJ column, black-mouthed form 500 mL, black-mouthed formumethanol (98: 2) 100 mL, black-mouthed form-methanol-water (85: 15: 1) 1000 DIL, black-mouthed form-methanol concentrated ammonia water ( The target fraction was collected by thin layer chromatography — (TLC) analysis as an index and concentrated to give a pale yellow color. 2.86 g (yield: 77.3%) of the target product was obtained as an oily substance.This compound was analyzed by TLC (silica gel plate, developing solvent: black-form-methanol-water (75: 25: 2)). As a result, a single spot was detected by detection with iodine, 0.2-ninhydrin reagent and 50% sulfuric acid-methanol reagent.

EI-MS (m / z): 370 (M ⁺ ) Reference Example 2. Synthesis of N-5, 8, 11, 14, 17-eicopentaenoylethylenediamine (EPA-EM)

Eicosapentaenoic acid (95% or more) (3.03 g (10rainol)) and N, N-carbonyldiimidazole (1.78 g) were dissolved in anhydrous THF (10 mL) and reacted at room temperature under a nitrogen stream for about 1 hour. Next, a solution prepared by dissolving 1.5 g (25 Dimol) of EDA and 2.55 g (25 mmol) of triethylamine in 10 mU of anhydrous THF was added to the reaction solution under ice-cooling, and the mixture was reacted for 2 hours. After completion of the reaction, 60 niL of 0.1N hydrochloric acid and 300 niL of chloroform-methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The resulting concentrate is placed on a silica gel (160 g) column, 1000 mL of black form-methanol-water (85: 15: l), and black form-methanol-concentrated aqueous ammonia (85: 15: 1). The eluate was flowed and eluted in the following order. The eluted fraction obtained was collected by thin layer chromatography (TLC) analysis as an index, and the target fraction was collected and concentrated.

2.72 g of the target substance (79% yield) was obtained by TLC analysis (silica gel plate, eluent: chloroform-methanol-water (75: 25: 2)). A single spot was detected by detection with the 0.2¾ ninhydrin reagent and 50% sulfuric acid-methanol reagent.

 5 EI-MS (πι / ζ): 344 (M +) Reference Example 3. Synthesis of N-9,12-octadecadienolethylenediamine (LA-ED A)

 2.8 g (10 mmol) of linoleic acid (95% or more) and 1.78 g (ll mmol) of Ν, Ν-carbodildimidazole were dissolved in anhydrous THF 10 mL, and reacted at room temperature under a nitrogen stream for about 1 hour. Then, a solution of 1.5 g (25 mmol) of EM and 2.55 g (25 mol) of triethylamine dissolved in 10 mL of anhydrous THF was added to the reaction mixture under ice-cooling, and the mixture was reacted for 2 hours. After completion, 60 mL of 0.1N hydrochloric acid and 300 mL of mouth methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The resulting concentrate is placed on a column of gel (160 g), and the form of methanol-water (85: 15: l) (1000 mL) and the form-methanol-aqueous ammonia (85: 15: 1) are used in this order. The eluate was run and eluted. The target fractions were collected from the obtained eluted fractions using thin layer chromatography (TLC) analysis as an index, and concentrated to obtain 2.430 g (yield: 75.5%) of the target product. The substance was analyzed by TLC (silica gel plate, developing solvent: black form-methanol-aqueous-water (75: 25: 2)), which was detected by iodine, 0.2-ninhydrin reagent and 50-methanol sulfate-methanol reagent. A single spot was shown.

EI-MS (Di / z): 322 ( ⁺ ) Example 1. N- [3- (3,4-dihydroxyphenylpropyl): -X '(4,7,10,13,16,19-docosahexaenoyl) ethylenediamine (DHA-ED Synthesis of (A-CA)

6!¾)を得た。 なお、 本物質は、 TLC分析（シリ力ゲルプレー 卜、 展開溶媒： クロ口ホルム一メタノー ルー水（ 85:15:1))を行ったところ、 沃素、 紫外線及び 50¾硫酸—メ 夕ノール試薬による検出で単一スポッ トを示した。 709 mg (3.94 mmolj and Ν'-hydroxysuccinimide (hereinafter abbreviated as HOSu)) of 543 mg (4.73 mmol) of 3,4-dihydroxycinnamic acid (capionic acid) in 3 mL of N-dimethylformamide and 5 mL of dichloromethane Then, 903 mg (4.73 mmol) of 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (hereinafter abbreviated as WSC) was added, and the mixture was reacted for 2 hours. A solution prepared by dissolving 1.457 g (3.94 mmol) of DHA-EDA obtained in Reference Example 1 and 482 mg (4.73 ramol) of triethylamine in 5 mL of dichloromethane was added under ice-cooling, and reacted at room temperature for 6 hours under a nitrogen stream. After completion of the reaction, 60 mL of 0.1N hydrochloric acid and 300 mL of chloroform-methanol (2: 1) were added to the reaction solution, the layers were separated, and the lower layer was separated and concentrated under reduced pressure. Silica gel dissolved in black-mouthed form and previously activated with black-mouthed form 100 g), and eluted with 500 mL of black-mouthed form, 500 DIL of black-mouthed formum (98: 2), 500 mL of black-mouthed form-methanol (95: 5) in the order of 500 mL, and eluted. The target fractions were collected from the fractions using thin layer chromatography (TLC) analysis as an index, and the target fractions were concentrated.

6! ¾). The substance was subjected to TLC analysis (silica gel plate, developing solvent: black-mouthed form-methanol solution (85: 15: 1)), which was detected with iodine, ultraviolet light and 50¾ sulfuric acid-methanol reagent. Indicates a single spot.

'HN R (400Hz, C ₅ D ₅ N): δ 0.9Κ3Η, t, -CH ₃ ), 2.03C2H, m, -CH ₂ CH ₃ ;.

2.16 (2H, t, CH _z CH ₂ C0NH), 2.27C2H, m,-CH = CHCH ₂ CH = CHCH ₂ CH ₂ , 2.79 (10H, m, -CH = CHCH 2 CH = CH-), 3.20~3.50 (4H, m, -NHCH Z CH Z NH- j,

5.33 (12H, m, -CH = CHCH ₂ CH = CH-), 6.35 (lH, d, -CH = CHC ₆ H ₃ (0H) ₂ ),

6.90 to 7.10 (3H, t, -C ₆ H ₃ (0HJ ₂ ), 7.42 (1H, d, -ΟΗ = αθ ₆ Η ₃ (ΟΗ) ₂ ), 8.09 (1H, t, -C0NHCH ₂ CH _2- ) , 8.35 (1H, t, -NHCOCH ^ CH-). Example 2. X- [3- (3,4-dihydroxyphenylpropyl)]-N '-(5, 8, 11, 14) Of 17,17-Ecopeneenoyl) ethylenediamine (EPA-EDA-CA)

 After dissolving 783 mg (4.35 mniol) of 3,4-dihydroxycinnamate (capionic acid) and 600 mg of HOSu (5.22 mmol) in 3 mL of N, N-dimethylformamide and 5 mL of dichloromethane, W997 mg (5.22 ol) Was added and reacted for 2 hours. Then, a solution of 1.497 g (4.35 ol) of EPA-EDA obtained in Reference Example 2 and 532 mg (5.22 mmol) of triethylamine dissolved in 5 mL of dichloromethane at 5 mL of dichloromethane was added to the reaction mixture under ice-cooling, and the mixture was added under a nitrogen stream. The reaction was performed at room temperature for 6 hours. After completion of the reaction, 60 mL of 0.1N hydrochloric acid and 300 mL of chloroform-formanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The obtained concentrate was dissolved in a small amount of chloroform at a port, applied to a silica gel (100 g) column, and 482 mg (yield: 20.9%) of the desired product was obtained from a fraction eluted at a port-form-methanol (95: 5). . When this substance was analyzed by TLC (silica gel plate, developing solvent: black form-methanol-water-aqueous (85: 15: 1)), it was detected by iodine, ultraviolet light and 50¾ sulfuric acid-methanol reagent. One spot was shown.

_{1H-NR (400Hz, C 5} D5N ;: δ 0.92 (3H, t, -CH 3),

1.56C2H, Di, -CH ₂ CH = CHCH ₂ CH _2- ),

2.05 (4H, m, -CH ₂ CH ₃ , -CH ₂ CH ₂ CH ₂ CONH-), _{2.1K2H, m, -CH 2 C0NH -} ), 2.80 (8H, m, -CH = CHCH 2 CH = CH-),

3.30 to 3.50 (4H, ni, -NHCH ₂ CH ₂ NH-) _t 5.33C10H, m, CH = CHCH _Z CH = CH-),

6.35 (1H, d, -CH = CHC ₆ H ₃ (0H) ₂ ), 6.90-7.10 (3H, t, -C ₆ H ₃ (0H) ₂ j,

7.42 (1H, d, -CH = CHC _fe H ₃ (0H) ₂ ), 8.09 (1H, t, -C0NHCH ₂ CH _2- ),

 8.35 (1H, t, -NHC0CH-CH-). Example 3. Ν '-[3- (3,4-dihydroxyphenylpropyl)]---(9,12-octadecadienoy) 3) Synthesis of ethylenediamine (LA-EDA-CA) 3,4-dihydroxycinnamic acid (cafic acid) 396oig (2.2mol) and HOSu 304mg (2.6ramol) were mixed with 3mL of N, N-dimethylformamide and dichloromethane. After dissolution in 5 mL of methane, WSC 504rag (2.6 niiol) was added and reacted for 2 hours. Then, a solution of 718 mg (2.2 mmol) of LA-EDA obtained in Reference Example 3 and 269 mg (2.64 mmol) of triethylamine dissolved in 5 mL of anhydrous THF was added to the reaction mixture under ice-cooling, and the mixture was stirred at room temperature under a nitrogen stream. For 6 hours. After completion, 60 mL of 0.1N hydrochloric acid and 300 mL of chloroform-methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The obtained concentrate was dissolved in a small amount of chloroform and applied to a silica gel UOOg) column, and 380 mg (yield: 39¾) of the desired product was obtained from a fraction eluted with chloroform-methanol (95: 5). The substance was analyzed by TLC (silica gel plate, eluent: chloroform-form-methanol-aqueous solution (85: 15: 1)) and detected with iodine, ultraviolet light and 50% sulfuric acid-methanol reagent. It showed a single spot.

¹ HN R (400Hz _> C ₅ D ₅ N ;: δ 0.85 (3H, t, -CH ₃ j,

1.23 to 1.26 (16Η, πι, -CH ₂ -λ 1.49C2H, t, -CH _z C0 Hj,

2.00C4H, in,-CH = CHCH ₂ CH = CHCH _2- ), 2.73 (2H, DI, -CH = CHCH ₂ CH = CH 3.30 ~ 3.50C4H, m, -NHCH ₂ CH ₂ NH-), 5.3K4H, m, -CH = CH-),

6.35C1H, d, -CH = CHC _i> H ₃ (0H ₂ ), 6.90 to 7.10C3H, t, -C _t H ₃ (0H) ₂ ),

7.42C1H, d, -CH = CHC ₆ H ₃ (0H) ₂ ), 8.09 (1H, t, -C0NHCH ₂ CH _2- ),

 8.35 (1H, t, -NHC0CH-CH-). Example 4. N- [3- (4-hydroxy-3-methoxyphenylpropyl)]-Ν '-(4,7,10,13,13) Synthesis of 16, 19-docosahexaenoyl) ethylenediamine (DHA-EDA-HMCA)

 After dissolving 524 mg (2.7 mmol) of 4-hydroxy-3-methoxycinnamic acid (ferulic acid) and 372 mg (3.24πιπιο1) of HOSu in 3 mL of N, N-dimethylformamide and 5 mL of dichloromethane, 619 mg of WSC (3.24 O1) was added and reacted for 2 hours. Next, a solution of 986 mg (2.7 vol) of DHA-EDA obtained in Reference Example 1 and 330 mg (3.24ππηο1) of triethylamine dissolved in 5 mL of dichloromethane was added to the reaction mixture under ice-cooling, and the mixture was added under a nitrogen stream. The reaction was carried out at room temperature for 4 hours. After completion of the reaction, 60 mL of 0.1N hydrochloric acid and 300 niL of chloroform (2: l) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The obtained concentrate was loaded on a silica gel (100 g) column, and the desired product 775 (yield: 53.2 得) was obtained from the fraction eluted with chloroformumethanol (98: 2). Developing solvent: black-mouthed form-methanol-aqueous (85: 15: 1)) showed a single spot by detection with iodine, ultraviolet light and 50¾ sulfuric acid-methanol reagent.

¹ HN R (400Hz, C ₅ D ₅ N): δ 0.9Κ3Η, t, -CH ₃ ), 2.03C2H, m, -CH ₂ CH ₃ ), 2.10 (2H, t, CH ₂ CH ₂ CONH-J, 2.25C2H, m, -CH = CHCH ₂ CH = CHCH ₂ CH ₂ ), 2.79 (10H, ni, -CH = CHCH ₂ CH = CH-), 3.10 ~ 3.40C4H, m, -NHCH ₂ CH ₂ NH-) , _{3.79 (3H, s, -C 6} H 3 (0CH 3) 0H), 5.33C12H, m, -CH-CHCH 2 CH = CH-), 6.25 (1H, d, -CH = CHC 6 H 3 (0CH 3 ) 0H),

6.79 to 7.00 (3H, t, -C ₆ j { ₃ (0CH ₃ ) 0iO.

7.30 (1H, d, -CH = CHC ₆ H ₃ (0CH ₃ ) 0H),

7.80C1H, t, - HC0CH = CH- ), 7.95 (1H, t, -C0 HCH 2 CH z -).

 In addition, EPA-EDA-HMCA, LA-EM-HMCA and LA-DET-HMCA were similarly synthesized using EPA-EDA, LA-EDA and LA-DET, respectively, instead of MA-EDA. Example 5 N- [3- (3,4-Dimethoxyphenylpropyl)]-N '-(4,7,10,13,16,19-docosahexaenoyl) ethylenediamine (MA- EDA '' MCA) Synthesis

After dissolving 479 mg (2.3 mmol) of 3,4-dimethyloxycinnamic acid and 317 mg (2.76 mmol) of HOSu in 3 mL of N, N-dimethylformamide and 5 mL of dichloromethane, add 527 mg (2.76niraol) of WSC and react for 2 hours I let it. Next, a solution of A-EDA 828rag (2.24 mmol) obtained in Reference Example 1 and 274 mg (2.7 ramol) of triethylamine dissolved in 5 mL of dichloromethane was added to the reaction mixture under ice-cooling, and the mixture was cooled to room temperature under a nitrogen stream. For 3 hours. After completion, 60 mL of 0.1N hydrochloric acid and 300 mL of chloroform-methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The obtained concentrate was applied to a silica gel (60 g) column, and 923 rag (yield: 73.6%) was obtained from a fraction eluted with chloroform (98: 2). The substance was analyzed by TLC (silica gel plate, developing solvent: chloroform-form methanol-water (85: 15: 1)), and was analyzed using iodine, ultraviolet rays, and 50% sulfuric acid-methanol reagent. Detection showed a single spot. 'H-NMR (400Hz, C 5 D 5 NJ: δ 0.9K3H, t, -CH 3), 2.03C2H, m, -CH 2 CH 3 J, 2.16 (2H, t, -CH 2 CH 2 CONH-j , 2.27 (2H, m, CH = CHCH ₂ CH = CHCH ₂ CH _2-

2.79 (10H, m, -CH = CHCH 2 CH = CH-), 3.20~3.50 (4H, m, -NHCH 2 CH Z NH-),

3.79C6H, s, -C ₆ H ₃ (0CH ₃ ; ₂ ;, 5.33 (12H, m, -CH = CHCH ₂ CH = CH-),

6.5K1H, d, -CH = CHC ₆ H ₃ (OCH ₃ ) ₂ ), 6.95 to 7.15 (3H, t, -C ₆ ] | ₃ (0CH ₃ ) ₂ ),

7.4K1H, d,-= CHC ₆ H ₃ (0CH ₃ ) ₂ ), 8.16C1H, t, -C0NHCH ₂ CH _2- ),

8.39C1H, t, -NHC0CH = CH-).

 EPA-EDA-MCA and LA-EM-MCA were also synthesized using EPA-EM and LA-EM, respectively, instead of DHA-EDA. Reference Example 4. Synthesis of N-9,12-octadecadienoljethylentriamine (LA-DET)

Linoleic acid (95% or more) 2.8 g (10rainol) and N, N-carbodildimidazole 1.78 g (llmmol) were dissolved in anhydrous THF (10 mL), and reacted at room temperature under a nitrogen stream for about 1 hour. Next, a solution prepared by dissolving 2.06 g of diethylenetriamine (hereinafter abbreviated as DET) and 2.06 g of triethylamine in 10 mL of anhydrous THF was added to the reaction solution under ice-cooling, and the reaction was carried out for 4 hours. After completion of the reaction, 60 mL of 0.1 N hydrochloric acid and 300 πL of chloroform-methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The resulting concentrated solution is applied to a silica gel (160 g) column, and the mixture is treated with black-mouthed form, black-mouthed methanol (95: 5), black-mouthed form-methanol-water (80: 20: 1), chloroform-methanol The eluate was eluted by flowing the eluate in the order of concentrated aqueous ammonia (80: 20: 1). The eluted fraction obtained was collected by thin layer chromatography (TL analysis as an index, and the target fraction was collected and concentrated to give 1.78 g (yield 48%) of the target product as a pale yellow oil. The substance is analyzed by TLC (Silica gel plate, developing solvent: black-mouthed form-methanol mono-concentrated ammonia water (80: 20: 1)), the single detection by iodine, 0.2% ninhydrin reagent and 50% sulfuric acid-methanol reagent Spotted.

EI-S (m / z): 365 (M +) Reference Example 5. Synthesis of N-5,8, ll, 14,17-eicopentaenoylethylethylenetriamine (synthesis of EPA-DEO)

Eicosapentaenoic acid (95% or more) 3.03 g (10 related ol) and N, N-carbonyldiimidazole ^ ^^! ^ Was dissolved in ¹ ! ^ 10 mL, and reacted at room temperature under a nitrogen stream for about 1 hour. Next, a solution of 2.06 g (20 mmol) of DET and 2.06 g of triethylamine dissolved in 10 mL of anhydrous THF was added to the reaction mixture under ice-cooling, and the mixture was reacted for 4 hours. After completion of the reaction, 60 mL of 0.1 N hydrochloric acid (60 mL) and 300 niL of form-methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The obtained concentrate is applied to a silica gel (160 g) column, and the mixture is concentrated in form: black form, black form-methanol (95: 5), black form-methanol-water (80: 20: 1), chloroform-methano, 1-ru The eluate was eluted by flowing an eluate in the order of aqueous ammonia (80: 20: 1). The target fraction was collected from the eluted fraction obtained using TLC analysis as an index, and concentrated to give 1.85 g (yield 47.8 mg) of the target product as a pale yellow oily substance. Force gel plate, developing solvent: black-mouthed form-methanol-concentrated aqueous ammonia (80: 20: 1)), a single spot was detected with iodine, 0.2-ninhydrin reagent and 50% sulfuric acid-methanol reagent showed that.

EI-MS (m / z): 387 (M +) Reference Example 6. Synthesis of N-4, 7, 10, 13, 16, 19-docosahexaenoyl acetylene triamine (DHA-DET)

 3.28 g (10raniol) of docosahexanoic acid (95% or more) and 1.78 g (llniraol) of N, N-carbonyldiimidazole were dissolved in anhydrous THF 10 mL, and reacted at room temperature under a nitrogen stream for about 1 hour. Next, a solution prepared by dissolving 2.06 g (20 mmol) of DET and 2.06 g of triethylamine in 10 mU of anhydrous THF was added to the reaction mixture under ice-cooling, and the mixture was reacted for 4 hours. After completion of the reaction, 60 mL of 0.1N hydrochloric acid and 300 mL of formaldehyde methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The resulting concentrate is applied to a silica gel (160 g) column, and the following are used: kuroguchi form, kuroguchi form-methanol (95: 5), kuroguchi formuminol-monoethanol (80: 20: 1), kuroguchi form-methanoine The eluate was eluted by flowing the eluate in the order of roux-concentrated aqueous ammonia (80: 20: 1). The target fraction was collected from the obtained eluted fraction using TLC analysis as an index, and concentrated to obtain 2.06 g (yield: 50.0¾) of the target product as a pale yellow oil. The substance was analyzed by TLC (silica gel plate, developing solvent: black-mouthed formum alcohol-concentrated aqueous ammonia (80: 20: 1)), using iodine, 0.2% ninhydrin reagent and 50% sulfuric acid-methanol reagent. Detection showed a single spot.

EI-MS (m / z): 413 ( ⁺ ) Example 6. N- [3- (3,4-dimethoxyphenylpropyl)]-(9,12-octadecadienol) Synthesis of diethylenetriamine (LA-DET-DMCA)

Dissolve 424 mg (2.04 DIDIO1) of 3,4-dimethyloxycinnamate and 396 mg (2.4 mmol) of N, N ^: -carbonyldiimidazole in 10 mU of anhydrous THF. The reaction was carried out at room temperature for 2 hours. Next, a solution prepared by dissolving 746 mg (2.04 mmol) of LA-DET obtained above and 208 mg (2.64 mmol) of triethylamine in 10 mL of anhydrous THF was added to the reaction solution, and reacted for about 4 hours. After completion of the reaction, 60 mL of 0.1N hydrochloric acid and 300 mL of chloroform-methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The obtained concentrate was loaded on a silica gel (60 g) column, and eluted with chloroform-methanol-acetic acid (85: 15: 1). The target fraction was collected from the obtained eluted fraction using TLC analysis as an index, and concentrated to obtain 74 mg (yield 45.6%) of the target product as a pale white powder. This substance was analyzed by TLC (silica gel plate, developing solvent: black form-methanol-water (85: 15: 1)) and detected by iodine, ultraviolet light and 50% sulfuric acid-methanol reagent. Spots were shown.

'H-NMR (400 Hz, C ₅ D ₅ N): δ 0.85C3H, t, -CH ₃ ),

1.23-1.26 (16Η, πι,-CH _2- ), 1.49 (2Η, t, -CH ₂ C0NH-),

2.00 (4H, m, -CH = CHCH ₂ CH = CHCH _2- ), 2.73 (2H, m, -CH = CHCH ₂ CH = CH-),

2.94 to 3.00 (4H, t, CH ₂ NHCH _2- ),

3.31 to 3.43 (4H, m, -NHCH ₂ CH ₂ NHCH ₂ CH ₂ NH-),

3.78 (6H, s, -C ₆ H ₃ (0CH ₃ ) ₂ ), 5.3K4H, m, -CH = CH-),

6.5K1H, d, -CH = CHC ₆ H ₃ (0CH ₃ ) ₂ ), 6.95 to 7.15 (3H, t, -C ₆ H ₃ (0CH ₃ ) ₂ ), 7.40 (1H, d, -CH = CHC ₆ H ₃ (OCH ₃ ) ₂ ), 8.06C1H, t, -C0NHCH ₂ CH _2- ),

8.33 (1H, t, -NHC0CH = CH-). Example 7. N- [3- (3,4-Dimethoxyphenylpropenyl)]-X,-(5, 8, 11, 14, Synthesis of 17-eicopenylenol) diethylenetriamine (EPA-DET-DMCA) 362 mg (l.74ππηο1) of 3,4-dimethoxycinnamic acid and 338 mg (2.08 mniol) of N, N-carbonyldiimidazole were dissolved in anhydrous THF 10 mL, and reacted at room temperature under a nitrogen stream for 2 hours. Next, a solution prepared by dissolving 674 mg (1.74 mmol) of EPA-DET obtained above and 177 mg of triethylamine in 10 mU of anhydrous THF was added to the reaction solution, and the mixture was reacted for about 4 hours. After completion of the reaction, 60 mL of 0.1N hydrochloric acid and 300 mL of chloroform-methanol (2: 1) were added to the reaction solution, and the mixture was separated. The lower layer was separated and concentrated under reduced pressure. The obtained concentrate was loaded on a silica gel (60 g) column, and eluted with black form-methanol-acetic acid (85: 1 5: 1). The target fractions were collected from the obtained eluted fractions using TLC analysis as an index and concentrated to obtain 440 mg (yield: 43.8%) of the target compound as a pale white powder. The substance was analyzed by TLC (silica gel plate, developing solvent: black form-methanol / aqueous (85: 15: 1)) and detected by iodine, ultraviolet light, and 50% sulfuric acid-methanol reagent. Spotted.

_{1H-NMR (400Hz, C 5} D 5 N): δ 0.92C3H, t, -CH 3),

1.56 (2H, m, -CH ₂ CH = CHCH _Z CH _Z- ),

2.05C4H, m, -CH ₂ CH ₃ , -CH ₂ CH ₂ CH ₂ CONH-),

2.1K2H, t, -CHzCONH-), 2.80 (8H, m, -CH = CHCH 2 CH = CH),

2.97-3.02 (4H, m, -CH ₂ NHCH ₂ ),

_{3.34~3.36 (4H, m, -NHCH 2} CH 2 NHCH Z CH 2 NH),

3.79C6H, s, -C ₆ H ₃ (0CH ₃ ) ₂ ), 5.33 (10H, m, -CH = CHCH _Z CH = CH-),

6.5K1H, d, -CH = CHC ₆ H ₃ (OCH ₃ ) ₂ ), 6.95 to 7.15C3H, t, -C ₆ H ₃ (0CH ₃ ) ₂ ),

7.40 (1H, d, -CH = CHC ₆ H ₃ (OCH ₃ ) ₂ ), 8.09 (1H, t, -C0NHCH ₂ CH _2- ),

8.35 (1H, t, -NHC0CH = CH- ;. Example 8. N- [3- (3,4-dimethyloxypropyl)]-N "-(4,7,10,13,16,19-docosahexaenoyl) diethylenetriamine (DHA-DET-MCA) Synthesis of

 406 mg (l.94ππηο1) of 3,4-dimethoxycinnamate and 379 mg (2.34 mmol) of N, N-carbonyldiimidazole were dissolved in 10 mU of anhydrous THF and reacted at room temperature under a nitrogen stream for 2 hours. Next, a solution of 805 mg (l.95 mmol) of DHA-DET obtained above and 199 mg of triethylamine dissolved in 10 mL of anhydrous THF was added to the reaction solution, and the mixture was reacted for about 4 hours. After completion of the reaction, 60 mL of 0.1N hydrochloric acid and 300 mL of chloroform-methanol (2: 1) were added to the reaction solution, and the mixture was separated. The obtained concentrate was loaded on a silica gel (60 g) column, and eluted with chloroform-methanol-acetic acid (85: 1 5: 1). The eluted fraction was collected by TLC analysis as an index to collect the target fraction, which was concentrated to obtain 498 mg (yield: 42.5%) of the target compound as a pale yellow powder. The substance was analyzed by TLC (silica gel plate, eluent: chloroform-form-methanol-water (85: 15: 1)), and a single spot was detected using iodine, ultraviolet light and 50¾ sulfuric acid-methanol reagent. showed that.

¹ H-NMR (400 Hz, C ₅ D ₅ N): δ 0.9Κ3Η, t, -CH ₃ ), 2.03C2H, m, -CH _z CH ₃ j, 2.16 (2H, t, -CH _Z CH ₂ CONH- ), 2.27 (2H, III, CH = CHCH ₂ CH = CHCH ₂ CH ₂ , 2.79C10H, m, -CH = CHCH ₂ CH = CH-), 3.00 to 3.05C4H, t, -CH ₂ NHCH _2- ) ,

3.11 ~ 3.22C4H, m,-HCH _Z CH ₂ NHCH ₂ CH _Z NH),

3.79 (6H, s, -C ₆ H ₃ (0CH3) ₂ ) ₍ 5.33C12H, m, -CH = CHCH _Z CH = CH),

6.5K1H, d, -CH = CHC ₆ H ₃ (OCH ₃ ) ₂ ), 6.95 to 7.15 C3H, t, -C ₆ H ₃ (0CH ₃ ) ₂ j, 7.41 (lH ₍ d, -CH-CHC ₆ H ₃ (OCH ₃ ) ₂ ),

_{8.16 (1H, t, -C0NHCH 2} CH 2), 8.39C1H, t, -NHCOCH = CH-). Test Example 1. Measurement of human neutrophil o ₂ -production inhibitory activity

The human acute myeloid leukemia cell HL-60, fetal calf serum was 3 x 10 ⁵ cells / mL will by Uni suspended in RPMI-1640 fold locations containing 10% and dimethicone Rusuruhokishido 1.4, C0 ₂ incubator _{(37 ° C, C0 2 5.0} %, humidity 100¾) were differentiated for 4 days culture to neutrophil-like in. Added differentiated human Bok neutrophils MEM medium Fuyunorure head no addition, was dispensed its 25 L in a 96-well play Bok to prepare cell concentration 2XLO ^b pieces / mL. The compounds of the present invention as a methanol solution of a predetermined concentration 200-fold, a material obtained by adding the same medium 200 This was incubated for 15 min at 25 L addition C0 ₂ incubator to the cell suspension. At this time, 25 L of the same medium was added to the sample-free group and the 100 ° inhibition control group.

After incubation for 15 min, the 100% inhibition control group was treated with 4 mg / mL saline solution of cytochrome C, 200 g / niL ethanol solution of TPA in l ^ L / mL and SOD 15000 L 7 mL (= 4.2 mg / mL) saline. solution 40 L / mL and those 50 added with the other in the Chitoku opening one厶C solution things that the TPA alone was added in the same concentration 50 / L was added, color was developed by 1 hr at C0 ₂ incubator . 550ηπι a plate reader (control, 570 nm) by measuring the absorbance of the 0 ₂ - to quantify the original cytochrome C instead produced by, 0 relative to the control group ₂ - from production inhibition rate 50% production inhibition concentration (IC ₅ Q ). Table 1 shows the results.

Table 10 ₂ -Production inhibitory activity Sample IC 5o (βg / ml)

L A-D E T-HMC A 4.4

 LA—D E T-DMC A 3.8

 E PA -DE T -DMCA 4.0

 DH A-D E T-DMCA 3.8

Test example 2. Measurement of phospholipase A ₂ inhibitory activity

As phospholipase A ₂ (PLA ₂ ), an 85 kDa cytoplasmic PLA ₂ (cPLA ₂ ) purified from egret platelets based on a previous report (FEBS Lett., 282, 326-330, 1991) was used. Was measured. The compound of the present invention was dissolved in methanol and used as a test solution. 1M DOO Lys - HCl (pH 9.0) was added to test solution and c P LA ₂ solution mixed-buffer 25 Les 50mM calcium chloride solution 20 L was a 200 L, and reacted at 37 hand 20 min. Then, as a substrate 1 Palmi toyl - 2- ^[l4 C] § La Kidonoiru - glycerin port phosphoethanolamine § Mi down the (0.5nmol / 50000dpm / 2 OO L ) was added and further reacted at 37 ° C 20 min . 1.25 mL of Dole reagent (isopropanol: heptane: IN sulfuric acid = 10: 40: 1) was added to stop the reaction, [ ^14C ] arachidonic acid fraction was collected by Dole's method, and its radioactivity was measured by liquid scintillation. The enzyme activity was measured by measuring with a counter. Table 2 shows the results. PLA ₂ inhibitory activity sample IC ₅ o (^ g / ml)

L A- EDA—DMC A 7.0

 DH A-ED A— DMCA 7.1

 DH A -EDA-HMC A 7.5

Test example 3. NGF production promoting activity against L-M cells

LM cells suspended in 199 medium containing 0.5% peptide at a concentration of 2 × 10 ⁴ / ml were inoculated into each well of a 96-well multiplate in an amount of 0.2 ml and cultured for 2-3 days. Thereafter, these cells were replaced with a test medium (199 medium containing 0.5% serum albumin) containing the compound of the present invention at various concentrations, and cultured for further 24 hours. After completion of the culture, the amount of NGF produced by the LM cells and released into the culture supernatant was measured by the enzyme immunoassay described below.

[Method of measuring NGF]

 Polystyrene 96-well multi-plate (MS manufactured by Sumitomo BeiClient)

-3496F) and an anti-mouse beta NGF polyclonal antibody (NGF prepared from mouse submandibular gland), which was prepared by the inventors using NGF as an antigen according to a standard method. LS. Furukawa, I. amo, Y. Furukawa. I Akazawa, E. Satoyoshi K. Itoh and K. Hayashi., J. Neurochem., 40, 734-744 (1983) =) (pH8.3) Left at C for 4 hours. Ma After removing the antibody not adsorbed to the well plate, each well was washed three times with a washing solution. Dispense 40 L of the standard beta-NGF (manufactured by Toyobo) solution or the culture supernatant of L-1 M cells obtained in the above experiment into each well, leave at 4 ° C for 18 hours, and then use the standard solution or The sample solution was removed ₍ each well was further washed three times).

 A solution (40 mU / nil, pH7.6) of beta-galactosidase-labeled anti-beta NGF monoclonal antibody (Boehringer Mannheim) was dispensed into each well in 50 portions, and allowed to stand at 37 ° C for 4 hours. It was removed and each hole was washed three times in the same manner as above. A solution of 4-methylumberifuryl-β-ϋ-galactoside (Sigma) (20 g / ml, pH 7.6) was dispensed into each well in a lOO ^ L, and allowed to react at room temperature for 1.5 hours. The enzyme reaction was stopped by dispensing 0.2N glycine sodium hydroxide buffer (pHIO, 3) into each well in 100 L portions, and the fluorescence intensity of the generated 4-methylumberifferon was measured with a plate reader. The NGF amount was calculated from the standard curve. Table 3 NGF production promotion activity Sample Production promotion activity

L A- EDA-C A About 10 times at 22 g / mL

 E PA-EDA-CA 6 g / mL about 6 times

DH A-EDA-CA about 4 times at 3 g / mL Industrial applicability

N- Ashiru N- substituted cinnamoyl ethylenedioxy § Mi emissions derivative conductor of the present invention, 0 ₂ arsenide Bok neutrophil - having production inhibitory activity, intracellular phospholipase eight ₂ inhibitory activity, pharmaceutical such as an anti-inflammatory agent It can be used as a therapeutic agent for dementia, spinal cord injury, peripheral nerve injury, diabetic neuropathy, amyotrophic lateral sclerosis, etc. because of its NGF production promoting action.

Claims

The following general formula, R'NH- CHzCHzNH r

N-acyl-N-substituted cinnamoylethylenediamine derivative represented by a- (wherein, R ¹ represents an acyl residue of a higher unsaturated fatty acid having 16 or more carbon atoms, and R ^z and R ³ independently represents a hydrogen atom or a lower alkyl group, and n is 1 or 2). Enclosure . 2 claim 1 0, and N Ashiru N- substituted cinnamoyl ethylenedioxy derivative as an active ingredient according ₂ - production inhibitor. 3. Claim 1 intracellular phospholipase A ₂ inhibitor as an active ingredient N Ashiru -N-substituted cinnamoyl ethylenedioxy Ryo amine derivative according.  4. A nerve growth factor production promoter comprising the N-acyl-N-substituted cinnamoylethylenediamine derivative according to claim 1 as an active ingredient.