JP6592802B2 - Phytosphingosine derivatives with sulfonyl - Google Patents

Description

  The present invention relates to a novel phytosphingosine derivative, particularly a phytosphingosine derivative having a sulfonyl.

Atopic dermatitis is a chronic pruritic skin disease with dry skin and severe itching, and repetitive remission and exacerbation.
The skin of a healthy person has a sufficient amount of moisturizing ingredients and lipids (such as sphingolipids) in the stratum corneum on the surface of the skin, and has a skin barrier function.
However, the skin of patients with atopic dermatitis has a lipid metabolism abnormality in which moisturizing components and sphingolipids, particularly ceramide, are reduced, and the skin is dried and the barrier function is reduced.
For this reason, foreign substances (allergens) and microorganisms that cause allergies are likely to invade, and the peripheral nerve extends to the vicinity of the stratum corneum in the outermost layer of the epidermis, and itching occurs with a slight stimulus, scratching the affected area.
By scratching, the barrier function is further destroyed, inflamed, and the itch becomes more vicious.

The treatment of atopic dermatitis has been mainly based on suppression of inflammation, and steroids and immunosuppressive drugs have been used.
Although such a drug can suppress inflammation, it cannot improve the decrease in barrier function, so that it is difficult to maintain the state after the inflammation has subsided, and the inflammation often recurs.

In atopic dermatitis mouse model skin, it is known that ceramide is decreased and sphingosylphosphorylcholine is increased compared to healthy mice, and that this substance is one of the endogenous itch factors (non-patented) Reference 1).
In a normal state, ceramide is produced from sphingomyelin by the action of sphingomyelinase.
However, in patients with atopic dermatitis, sphingomyelin deacylase is expressed and activated, and sphingomyelin is metabolized to sphingosylphosphorylcholine by deacylating the amide part instead of ceramide. This has led to a decrease in ceramide in the skin (Non-patent Document 2).

Experimental Dermatology, 20, 894-898 (2011) J. Invest. Dermatol., 115 (3), 406-413 (2000)

The present invention suppresses inflammation of atopic dermatitis and prevents recurrence by suppressing the action of lipid metabolizing enzyme deacylase, which reduces the amount of sphingolipid ceramide on the skin surface, which is one of the causes of the reduction of the barrier function of the skin. The aim is to develop new types of therapeutic agents.
On the other hand, human sphingomyelin deacylase has not been isolated and structure-determined, and no attempt has been made to use its inhibitor as a therapeutic drug for atopic dermatitis.

Therefore, we considered that inhibition of deacylase may increase ceramide production in addition to suppression of itchiness, and we considered a transition state model of deacylase and synthesized the model compound.
It was assumed that the transition state in the hydrolysis of the amide bond of sphingomyelin and glucosylceramide by deacylase is a hydrate with water added to the amide bond.
This differs from the substrate amide in that the amide carbon changes from a planar structure to a tetrahedral structure.
Therefore, we thought that a structure with a tetrahedral structure next to the nitrogen atom could be the transition state model.
Moreover, considering the adaptation to an external medicine for skin, the 1-position hydroxy group of sphingosine was not a water-soluble phosphate ester found in sphingomyelin, but a highly lipid-soluble substituent.
As a result, a phytosphingosine derivative having a sulfonyl as a therapeutic agent for atopic dermatitis was found and the present invention was completed.
Hereinafter, the present invention will be described in detail.

In the present invention, unless otherwise specified, an alkyl group is linear or branched having 1 to 6 carbon atoms such as methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, heptyl and the like. A linear alkyl group having 7 to 15 carbon atoms such as heptyl, octyl, decatyl, tridecylpentadecyl, etc .; an acyl group is a linear or branched C _2-6 such as acetyl, propionyl, isobutyryl, etc. Alkyl-CO-group means each.

「式中、Ｒ^１は、水素原子またはアルキル基を；Ｒ^２はアルキル基を、Ｒ^３およびＲ^４は、同一または異なって水素原子またはアシル基を、Ｒ^５は、アルキル基を、それぞれ意味する。」The first invention of the present invention is a phytosphingosine derivative represented by the following general formula [1].

“Wherein R ¹ represents a hydrogen atom or an alkyl group; R ² represents an alkyl group, R ³ and R ⁴ are the same or different and represent a hydrogen atom or an acyl group, and R ⁵ represents an alkyl group, respectively. To do. "

「式中、Ｒ^１ａは、炭素数１〜６の直鎖状または分岐状アルキル基、Ｒ^５ａは、炭素数７〜１５直鎖状のアルキル基を、それぞれ意味する。」A more preferable compound is a phytosphingosine derivative represented by the following general formula [1a].

“In the formula, R ^1a represents a linear or branched alkyl group having 1 to 6 carbon atoms, and R ^5a represents a linear alkyl group having 7 to 15 carbon atoms.”

  In the compounds of the general formulas [1] and [1a], when isomers (for example, optical isomers, geometric isomers, tautomers, etc.) exist, the present invention includes these isomers, and , Solvates, hydrates and crystals of various shapes.

  The second invention of the present invention is a therapeutic agent for atopic dermatitis comprising the phytosphingosine derivative represented by the above general formula [1] or [1a] as an active ingredient.

Since the phytosphingosine derivative of the present invention has an inhibitory action on an enzyme (SCDase) known to cause deacylation of sphingomyelin, it is presumed to exhibit the inhibitory activity of sphingomyelin deacylase. The
Moreover, since the phytosphingosine derivative of the present invention significantly suppresses scratching behavior in atopic dermatitis model mice, it is useful as a therapeutic agent for atopic dermatitis.

It is a figure which shows the result of the application | coating test with an atopic dermatitis model mouse. In the figure, ● represents a test compound, and ○ represents a control.

The phytosphingosine derivative of the general formula [1] can be produced by combining known methods, and for example, can be produced by the following method.

“Wherein R ^{1 to} R ⁵ have the same meaning as described above.”

(1) The compound of general formula [4] can be produced by reacting the compound of general formula [2] with the compound of general formula [3].
This reaction may be performed in an ether solvent such as tetrahydrofuran in the presence of a base such as triethylamine.
The reaction temperature may be appropriately determined depending on the solvent used, but is 20 ° C. to 60 ° C., preferably room temperature.

(2) The compound of general formula [1] can be produced by reacting the compound of general formula [4] with the compound of general formula [5].
This reaction may be performed in an ether solvent such as tetrahydrofuran in the presence of a base such as triethylamine.
The reaction temperature may be appropriately determined depending on the solvent used, but is 20 ° C. to 60 ° C., preferably room temperature.

In the compound represented by the general formula [4] in the production method described above, when isomers (for example, optical isomers, geometric isomers, tautomers, etc.) exist, these isomers can be used. Also, solvates, hydrates and various forms of crystals can be used.
Further, after completion of the reaction, the target product may be used as it is in the next reaction without isolation.

  The phytosphingosine derivative of the general formula [1] is combined with excipients, adjuvants, additives and the like, and various preparations such as solutions, suspensions, syrups, elixirs, extracts, powders, granules, Fine granules, tablets, capsules, liquids, gels, creams, lotions, mists, aerosols, foams, and aerosols can be used.

The therapeutic agent for atopic dermatitis comprising the phytosphingosine derivative of the general formula [1] as an active ingredient is used as a dosage form (solution, cream, ointment, gel, patch, aerosol, etc.) used for external preparations. It is preferable to use it.
Moreover, they can be manufactured by a conventional method.

For external preparations, water, lower alcohols, solubilizers, surfactants, emulsion stabilizers, gelling agents, adhesives, and other commonly used base components for obtaining desired dosage forms as necessary As needed, vasodilators, corticosteroids, keratolytic agents, moisturizers, bactericides, antioxidants, cooling agents, fragrances, pigments, etc., as long as the effects of the present invention are not impaired. Can be blended.
Hereinafter, the present invention will be described more specifically with reference to production examples and test examples, but the present invention is not limited thereto.

Production Example 1
<N-tosyl-1-octanesulfonyl phytosphingosine>

(1) A solution of tosyl chloride (1.45 g, 7.61 mmol) in tetrahydrofuran (8 mL) was added to a suspension of phytosphingosine (2.3 g, 7.24 mmol) and triethylamine (1.2 mL, 8.69 mmol) in tetrahydrofuran (50 mL) at room temperature. .
After stirring for 2 hours, the reaction mixture was diluted with ethyl acetate (50 mL), washed twice with 5% hydrochloric acid, washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was recrystallized (solvent: ethyl acetate / hexane) to obtain N-tosylphytosphingosine (2.95 g, 86%).

Melting point: 120.0-121.0 ° C
1H-NMR (400MHz, CD3OD) δ: 7.75 (2H, d, J = 8.2Hz), 7.36 (2H, d, J = 8.2Hz),
3.57-3.55 (2H, m), 3.48-3.39 (3H, m), 2.42 (3H, s), 1.37-1.21 (26H, m),
0.89 (3H, t, J = 6.4Hz)
13C-NMR (100MHz, CD3OD) δ: 144.5, 140.1, 130.7, 128.1, 76.8, 73.2, 61.8, 57.0,
33.7, 33.1, 30.9, 30.8, 30.5, 26.7, 23.7, 21.5, 14.4.

(2) A solution of 1-octanesulfonyl chloride (734 mg, 7.61 mmol) in tetrahydrofuran (6.3 mL) was added to a solution of N-tosylphytosphingosine (1.55 g, 3.29 mmol) and triethylamine (0.55 mL, 3.94 mmol) in tetrahydrofuran (20 mL). Added at room temperature.
After stirring for 2 hours, the reaction mixture was diluted with ethyl acetate (30 mL), washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography and recrystallized (solvent: ethyl acetate / hexane) to obtain 714 mg (34%) of N-tosyl-1-octanesulfonylphytosphingosine (Compound 1).

Melting point: 106-106.5 ° C
1H-NMR (400MHz, CDCl3) δ: 7.77 (2H, d, J = 8.2Hz), 7.31 (2H, d, J = 8.3Hz),
4.40 (1H, dd, J = 4.1,10.5Hz), 4.24 (1H, dd, J = 2.7,11.0Hz), 3.65-3.61 (3H, m),
2.98 (2H, t, J = 7.3Hz), 2.43 (3H, s), 1.78 (2H, t J = 7.8Hz), 1.30-1.18 (36H, m),
0.90-0.86 (3H, m)
13C-NMR (100MHz, CDCl3) δ: 143.8, 137.3, 129.8, 127.2, 73.7, 72.4, 68.2, 54.4,
50.2, 32.0, 31.9, 31.7, 29.7, 29.4, 28.9, 28.1, 25.6, 23.2, 22.7, 22.6, 21.5,
14.1, 14.0.

Production Example 2
<N-tosyl-1-ethanesulfonyl phytosphingosine>

A solution of 1-ethanesulfonyl chloride (243 mg, 1.89 mmol) in tetrahydrofuran (4.4 mL) is added to a solution of N-tosyl-phytosphingosine (0.85 g, 1.80 mmol) and triethylamine (0.30 mL, 2.16 mmol) in tetrahydrofuran (10 mL) at room temperature. added.
After stirring for 4 hours, the reaction mixture was diluted with ethyl acetate (15 mL), washed twice with 5% hydrochloric acid, washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was recrystallized (solvent: ethyl acetate / hexane) to obtain 0.82 g (81%) of N-tosyl-1-ethanesulfonylphytosphingosine (Compound 2).

Melting point: 85.5-86.0 ° C
1H-NMR (400MHz, CDCl3) δ: 7.77 (2H, d, J = 8.7Hz), 7.34 (2H, d, J = 8.2Hz),
5.49 (1H, d, J = 7.8Hz), 4.41 (1H, dd, J = 4.6,11.0Hz), 4.26 (1H, dd, J = 2.7,10.5Hz),
3.66-3.59 (3H, m), 3.06 (2H, dd, J = 7.3,14.7Hz), 2.43 (3H, s),
1.34 (3H, dd, J = 7.3,7.8Hz), 1.32-1.11 (26H, m), 0.88 (3H, dd, J = 6.4,6.9Hz)
13C-NMR (100MHz, CDCl3) δ: 144.0, 137.0, 129.7, 127.1, 85.5, 74.2, 69.7, 54.9,
33.4, 31.9, 29.6, 25.6, 22.7, 21.6, 14.1.

Production Example 3
<N-tosyl-1-octanesulfonyl-3,4-diacetylphytosphingosine>

Acetic anhydride (0.073 mL, 0.77 mmol) was added to a solution of N-tosyl-1-octanesulfonylphytosphingosine (100 mg, 0.154 mmol) in pyridine (0.62 mL) at room temperature.
After stirring for 6 hours, pyridine was distilled off under reduced pressure, diluted with ethyl acetate (10 mL), washed successively with water and saturated brine, and dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure.
The residue was purified by silica gel column chromatography (20% ethyl acetate / hexane) to obtain 60 mg (53%) of N-tosyl-1-octanesulfonyl-3,4-diacetylphytosphingosine (Compound 3).

1H-NMR (400MHz, CDCl3) δ: 7.77 (2H, d, J = 8.2Hz), 7.34 (2H, d, J = 8.2Hz),
5.44 (1H, d, J = 9.2Hz), 4.97 (1H, dd, J = 3.2,7.3Hz), 4.89 (1H, dt, J = 3.2,9.6Hz),
4.14 (2H, d, J = 3.7Hz), 3.74 (1H, m), 3.04 (2H, t, J = 7.8Hz), 2.43 (3H, s), 2.06 (3H, s),
2.03 (3H, s), 1.80 (2H, t, J = 7.8Hz), 1.29-1.18 (36H, m), 0.90-0.86 (6H, m)
13C-NMR (100 MHz, CDCl3) δ: 170.8, 169.9, 143.9, 137.4, 129.9, 127.2, 72.5, 72.0,
66.3, 52.1, 50.5, 31.9, 29.7, 28.9, 28.1, 25.3, 23.1, 22.6, 14.0

Production Example 4
<N-tosyl-1-octanesulfonyl-4-acetylphytosphingosine>

(1) N-tosylphytosphingosine (500 mg, 1.06 mmol) was added to a DMF (8.5 mL) solution of bis (trifluoromethanesulfonic acid) di-tert-butylsilyl (560 mg, 1.27 mmol) at 0 ° C.
After stirring at 0 ° C. for 10 minutes and at room temperature for 15 minutes, triethylamine (0.44 mL, 3.18 mmol) was added and stirred for 10 minutes.
Saturated aqueous ammonium chloride solution (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL × 2).
The organic layer was washed successively with water and saturated brine, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (25% ethyl acetate / hexane) to obtain 592 mg (91%) of a protected silylene.

(2) A solution of acetic anhydride (28 mg, 0.30 mmol) in pyridine (0.1 mL) was added to a pyridine (0.1 mL) solution of a protected silylene (30 mg, 0.05 mmol) at room temperature.
After stirring for 24 hours, the mixture was diluted with methylene chloride (10 mL), washed successively with 5% aqueous citric acid solution, water and saturated brine, and dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (20% ethyl acetate / hexane) to obtain 18 mg (56%) of an acetylated product.

(3) Hydrogen fluoride · pyridine (4.7 mg, 0.17 mmol) was added to a solution of acetylated compound (73 mg, 0.11 mmol) in tetrahydrofuran (1.1 mL) at 0 ° C.
After stirring at room temperature for 20 minutes, a saturated aqueous sodium hydrogen carbonate solution (5 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (10 mL × 2).
The organic layer was washed successively with water and saturated brine, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (50% ethyl acetate / hexane) to obtain 48 mg (84%) of a diol.

(4) A solution of 1-octanesulfonyl chloride (14 mg, 0.067 mmol) in tetrahydrofuran (0.2 mL) was added to a solution of diol (33 mg, 0.064 mmol) and triethylamine (0.01 mL, 0.077 mmol) in tetrahydrofuran (0.3 mL) at room temperature. It was.
After stirring for 20 minutes, the reaction mixture was diluted with ethyl acetate (10 mL), washed successively with water and saturated brine, and dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (35% ethyl acetate / hexane) to obtain 36 mg (82%) of N-tosyl-1-octanesulfonyl-4-acetylphytosphingosine (Compound 4).

Melting point: 44.5-46.0 ° C
1H-NMR (400MHz, CDCl3) δ: 7.78 (d, J = 8.2Hz, 2H), 7.32 (d, J = 8.2Hz, 2H),
5.79 (d, J = 9.2Hz, 1H), 4.79 (d, J = 10.1Hz, 1H), 4.44 (dd, J = 10.1, 6.4 Hz, 1H),
4.21 (dd, J = 10.5,2.7Hz, 1H), 3.82 (m, 1H), 3.46 (m, 1H), 3.05 (dd, J = 16.0,7.8Hz, 2H),
2.95 (d, J = 5.5Hz, 1H), 2.43 (s, 3H), 2.10 (s, 3H), 1.828-1.750 (m, 2H), 1.40 (brs, 36H),
0.88 (dd, J = 13.2, 6.4Hz, 6H)
IR (KBr, cm-1) ν3485,3397,3303,2919,2850,1629,1469,1336,1159,1057

Production Example 5
<N-tosyl-1-pentanesulfonyl phytosphingosine>

(1) Sodium 1-pentanesulfonate is converted to 1-pentanesulfonic acid with 10% aqueous hydrochloric acid, and 1-pentanesulfonic acid (181 mg, 1.2 mmol) in dimethylformamide / hexane (1: 7,0.46 mL) is thionyl chloride (1: 7,0.46 mL). 0.15 mL, 2.0 mmol) was added dropwise, and the mixture was stirred at 70 ° C. for 1 hour, and then heated to reflux for 2 hours.
After returning to room temperature, the hexane layer was separated, washed with 5% aqueous sodium carbonate solution, and the aqueous layer was extracted with hexane.
The solvent was distilled off under reduced pressure to obtain 1-pentanesulfonyl chloride (87 mg, 42%).

(2) A solution of 1-pentanesulfonyl chloride (20 mg, 0.12 mmol) in tetrahydrofuran (0.3 mL) was added to a solution of N-tosyl-phytosphingosine (50 mg, 0.11 mmol) and triethylamine (0.018 mL, 0.13 mmol) in tetrahydrofuran (0.58 mL). At room temperature.
After stirring for 1.5 hours, the reaction mixture was diluted with ethyl acetate (5 mL), washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (40% ethyl acetate / hexane) to obtain 41 mg (61%) of N-tosyl-1-pentanesulfonylphytosphingosine (Compound 5).

Melting point: 77.5-78.5 ° C
1H-NMR (400 MHz, CDCl3) δ: 7.78 (2H, d, J = 8.0Hz), 7.32 (2H, d, J = 8.0Hz), 5.47 (1H, brs),
4.41 (1H, dd, J = 10.4,4.0Hz), 4.23 (1H, dd, J = 10.4,2.4Hz), 3.68-3.50 (3H, m),
3.01 (2H, t, J = 8.0Hz), 2.77 (1H, brs), 2.43 (3H, s), 2.01 (1H, brs),
1.78 (2H, tt, J = 8.0,7.6Hz), 1.42-1.30 (4H, m), 1.32-1.21 (26H, m),
0.89 (6H, tt, J = 7.6,7.0Hz)
13C-NMR (100 MHz, CDCl3) δ: 143.7, 137.3, 129.8, 127.2, 73.6, 72.4, 68.2, 50.1, 31.9, 30.18, 30.17, 29.69, 29.68, 29.67, 29.64, 29.62, 29.5, 29.4, 29.3, 25.7 , 25.6, 22.9, 22.7, 22.0, 21.5, 14.1, 13.7
IR (KBr, cm-1) ν3506,3407,3357,1470,1438,1351,1164

Production Example 6
<N-tosyl-1-heptanesulfonyl phytosphingosine>

(1) Sodium 1-heptanesulfonate is converted to 1-heptanesulfonic acid with 10% aqueous hydrochloric acid, and 1-heptanesulfonic acid (260 mg, 1.4 mmol) in dimethylformamide / hexane (1: 7,0.55 mL) is thionyl chloride ( 0.18 mL, 2.5 mmol) was added dropwise, and the mixture was stirred at 70 ° C. for 1 hour and then heated to reflux for 2 hours.
After returning to room temperature, the hexane layer was separated, washed with 5% aqueous sodium carbonate solution, and the aqueous layer was extracted with hexane.
The solvent was distilled off under reduced pressure to obtain 1-heptanesulfonyl chloride (241 mg, 84%).

(2) A solution of 1-heptanesulfonyl chloride (24 mg, 0.12 mmol) in tetrahydrofuran (0.3 mL) was added to a solution of N-tosyl-phytosphingosine (50 mg, 0.11 mmol) and triethylamine (0.018 mL, 0.13 mmol) in tetrahydrofuran (0.58 mL). At room temperature.
After stirring for 1 hour, the reaction mixture was diluted with ethyl acetate (5 mL), washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (40% ethyl acetate / hexane) to obtain 52 mg (74%) of N-tosyl-1-heptanesulfonyl phytosphingosine (Compound 6).

Melting point: 98.5-99.5 ° C
1H-NMR (400 MHz, CDCl3) δ: 7.78 (2H, d, J = 8.2Hz), 7.33 (2H, d, J = 8.2Hz),
5.32 (1H, d, J = 8.2Hz), 4.41 (1H, dd, J = 11.0,4.6Hz), 4.22 (1H, dd, J = 11.0,3.2Hz),
3.68-3.56 (3H, m), 3.01 (2H, t, J = 7.8Hz), 2.60 (1H, d, J = 6.0Hz), 2.44 (3H, s),
1.86 (1H, d, J = 5.5Hz), 1.77 (2H, tt, J = 7.8,7.4Hz), 1.42-1.35 (4H, m),
1.31-1.24 (30H, m), 0.89 (6H, dt, J = 6.9,5.1Hz)
13C-NMR (100 MHz, CDCl3) δ: 143.8, 137.2, 129.8, 127.2, 73.6, 72.4, 68.2, 54.4, 50.2, 32.09, 32.08, 31.9, 31.43, 31.42, 29.70, 29.67, 29.66, 29.65, 29.36, 29.35 , 28.11, 28.08, 25.62, 25.61, 23.2, 22.7, 22.5, 21.6, 14.1, 14.0
IR (KBr, cm-1) ν3460,3357,3265,1460,1441,1335,1324,1169

Production Example 7
<N-tosyl-1-nonanesulfonyl phytosphingosine>

(1) Sodium 1-nonanesulfonate is converted to 1-nonanesulfonic acid with 10% aqueous hydrochloric acid, and 1-nonanesulfonic acid (271 mg, 1.3 mmol) in dimethylformamide / hexane (1: 7,0.5 mL) is thionyl chloride (1: 7,0.5 mL). 0.16 mL, 2.52 mmol) was added dropwise, and the mixture was stirred at 70 ° C. for 1 hour, and then heated to reflux for 2 hours.
After returning to room temperature, the hexane layer was separated, washed with 5% aqueous sodium carbonate solution, and the aqueous layer was extracted with hexane.
The solvent was distilled off under reduced pressure to obtain 1-nonanesulfonyl chloride (267 mg, 91%).

(2) A solution of 1-nonanesulfonyl chloride (27 mg, 0.12 mmol) in tetrahydrofuran (0.3 mL) was added to a solution of N-tosyl-phytosphingosine (50 mg, 0.11 mmol) and triethylamine (0.018 mL, 0.13 mmol) in tetrahydrofuran (0.58 mL). At room temperature.
After stirring for 2 hours, the reaction mixture was diluted with ethyl acetate (5 mL), washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (40% ethyl acetate / hexane), recrystallized (solvent: ethyl acetate / hexane), and 41 mg (56 mg) of N-tosyl-1-nonanesulfonyl phytosphingosine (compound 7) was obtained. %)Obtained.

Melting point: 102.0-103.0 ° C
1H-NMR (400 MHz, CDCl3) δ: 7.78 (2H, d, J = 8.2Hz), 7.32 (2H, d, J = 8.2Hz), 5.45 (1H, brs), 4.40 (1H, dd, J = 11.0, 4.1Hz), 4.22 (1H, dd, J = 11.0,3.2Hz), 3.66-3.59 (3H, m),
3.01 (2H, t, J = 7.8Hz), 2.43 (3H, s), 2.05 (2H, brs), 1.78 (2H, tt, J = 8.2,7.8Hz),
1.43-1.35 (4H, m), 1.30-1.22 (34H, m), 0.88 (6H, dt, J = 6.9,1.4Hz)
13C-NMR (100 MHz, CDCl3) δ: 143.8, 137.2, 129.8, 127.2, 73.6, 72.4, 68.1, 54.4, 50.2, 32.1, 31.8, 29.69, 29.68, 29.66, 29.65, 29.63, 29.4, 29.23, 29.16, 29.14 , 29.0, 28.1, 25.6, 23.2, 22.7, 22.6, 21.5, 14.10, 14.07
IR (KBr, cm-1) ν3547,3465,3354,2918,2849,1470,1453,1440,1352,1164

Production Example 8
<Np-propylbenzenesulfonyl-1-octanesulfonyl phytosphingosine>

(1) A solution of p-propylbenzenesulfonyl chloride (1.07 mg, 0.49 mmol) in tetrahydrofuran (1.8 mL) is added to a suspension of phytosphingosine (150 mg, 0.47 mmol) and triethylamine (0.08 mL, 0.57 mmol) in tetrahydrofuran (2 mL). Added at room temperature.
After stirring for 2 hours, the reaction mixture was diluted with ethyl acetate (10 mL), washed twice with 5% hydrochloric acid, washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was recrystallized (solvent: ethyl acetate / hexane) to obtain Np-propylbenzenesulfonyl phytosphingosine (129 mg, 55%).

(2) A solution of 1-octanesulfonyl chloride (23 mg, 0.11 mmol) in tetrahydrofuran (0.3 mL) was added to Np-propylbenzenesulfonylphytosphingosine (50 mg, 0.10 mmol) and triethylamine (0.017 mL, 0.12 mmol) in tetrahydrofuran (0.5 mL). To the solution was added at room temperature.
After stirring for 5 hours, the reaction mixture was diluted with ethyl acetate (5 mL), washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (40% ethyl acetate / hexane) to obtain 46 mg (68%) of Np-propylbenzenesulfonyl-1-octanesulfonylphytosphingosine (Compound 8).

Melting point: 79.5-81.5 ° C
1H-NMR (400 MHz, CDCl3) δ: 7.80 (2H, d, J = 8.4Hz), 7.32 (2H, d, J = 8.4Hz),
5,49 (1H, t, J = 8.4Hz), 4.40 (1H, dd, J = 11.0,4.8Hz), 4.22 (1H, dd, J = 11.0,3.2Hz),
3.67-3.57 (3H, m), 3.01 (2H, t, J = 7.8Hz), 2.77 (1H, d, J = 6.8Hz), 2.66 (2H, t, J = 7.6Hz),
2.01 (1H, d, J = 5.6Hz), 1.78 (2H, tt, J = 8.4,7.8Hz), 1.66 (2H, qt, J = 7.8,7.6Hz),
1.40-1.37 (4H, m), 1.35-1.21 (32H, m), 0.95 (3H, t, J = 7.6Hz),
0.88 (3H, dt, J = 6.8,2.0Hz)
13C-NMR (100 MHz, CDCl3) δ: 148.4, 137.4, 129.2, 127.2, 73.7, 72.4, 68.2, 54.4, 50.2, 37.9, 32.10, 32.09, 32.08, 31.9, 31.7, 29.71, 29.69, 29.68, 29.66, 29.4 , 28.9, 28.1, 25.6, 24.1, 23.2, 22.7, 22.6, 14.1, 14.0, 13.7
IR (KBr, cm-1) ν3515,3326,3192,1469,1343,1162,1148

Production Example 9
<Np-pentylbenzenesulfonyl-1-octanesulfonylphytosphingosine>

(1) A solution of p-pentylbenzenesulfonyl chloride (1.21 mg, 0.49 mmol) in tetrahydrofuran (0.5 mL) is added to a suspension of phytosphingosine (150 mg, 0.47 mmol) and triethylamine (0.08 mL, 0.57 mmol) in tetrahydrofuran (3 mL). Added at room temperature.
After stirring for 1.5 hours, the reaction mixture was diluted with ethyl acetate (10 mL), washed twice with 5% hydrochloric acid, washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was recrystallized (solvent: ethyl acetate / hexane) to obtain Np-pentylbenzenesulfonyl phytosphingosine (158 mg, 63%).

(2) A solution of 1-octanesulfonyl chloride (22 mg, 0.105 mmol) in tetrahydrofuran (0.3 mL) was added to Np-pentylbenzenesulfonyl phytosphingosine (50 mg, 0.095 mmol) and triethylamine (0.016 mL, 0.114 mmol) in tetrahydrofuran (0.46 mL). To the solution was added at room temperature.
After stirring for 3 hours, the reaction mixture was diluted with ethyl acetate (5 mL), washed successively with water and saturated brine, and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure.
The residue was purified by silica gel column chromatography (35% ethyl acetate / hexane) to obtain 43 mg (64%) of Np-pentylbenzenesulfonyl-1-octanesulfonylphytosphingosine (Compound 9).

Melting point: 88.5-89.5 ° C
1H-NMR (400 MHz, CDCl3) δ: 7.79 (2H, d, J = 8.4Hz), 7.32 (2H, d, J = 8.4Hz),
5.37 (1H, d, J = 8.8Hz), 4.41 (1H, dd, J = 10.8,4.8Hz), 4.22 (1H, dd, J = 10.8,3.2Hz),
3.64-3.55 (3H, m), 3.01 (2H, t, J = 7.8Hz), 2.67 (2H, t, J = 8.0Hz), 2.64 (1H, d, J = 7.2Hz),
1.90 (1H, d, J = 5.6Hz), 1.78 (2H, tt, J = 8.0,7.6Hz), 1.63 (2H, tt, J = 7.6,7.8Hz),
1.34-1.31 (4H, m), 1.29-1.22 (38H, m), 1.21-0.86 (9H, m)
13C-NMR (100 MHz, CDCl3) δ: 148.7, 137.3, 129.2, 127.2, 73.7, 72.4, 68.1, 54.4, 50.2, 35.8, 32.16, 32.15, 31.9, 31.7, 31.41, 31.40, 30.7, 29.70, 29.68, 29.67 , 29.66, 29.65, 29.4, 28.95, 28.94, 28.1, 25.60, 25.59, 23.2, 22.7, 22.6, 22.4, 14.1, 14.04, 13.95
IR (KBr, cm-1) ν3515, 3323, 3190, 1469, 1343, 1162, 1151

Test example 1
Using an enzyme (SCDase) known to cause deacylation of sphingomyelin, it was evaluated by the inhibitory activity of SCDase on sphingomyelin deacylation.
0.05 μmol, 0.10 μmol, 0.15 μmol equivalent of the test compound [10 mg / mL (ethanol: acetone = 1: 1 (v / v))] was put in a plastic tube, and after water was removed, 0.8% TritonX-100 9.5 μL of a 50 mM sodium acetate buffer solution (pH = 6.0) and 0.5 μL of SCDase stock solution were added and reacted at 37 ° C. for 30 minutes.
In advance, 4 μL of sphingomyelin [10 mg / mL (chloroform: methanol = 1: 1 (v / v))] was placed in a plastic tube, the water was removed, the reaction mixture was added, and the mixture was reacted at 37 ° C. for 60 minutes.
Thereafter, the water was evaporated by an evaporator and dried.
After drying, 20 μL of methanol and 20 μL of chloroform were added, followed by centrifugation [4 ° C., 10,000 rpm, 5 minutes], 20 μL of the supernatant was taken, and water was removed again with an evaporator.
It was dissolved in 6 μL of an extract [chloroform: methanol = 2: 1 (v / v)] and spotted 6 times at 0.5 μL on TLC aluminum sheet silica gel 60 (Merck).
As standard products, sphingomyelin [10 mg / mL (chloroform: methanol = 1: 1 (v / v))] and sphingosylphosphosylcholine [10 mg / mL (chloroform: methanol = 1: 1 (v / v))] Was spotted on the same aluminum sheet.
After that, develop with a developing solution [chloroform: methanol: acetic acid: water = 50: 30: 8: 7 (v / v / v / v))] and dry, then add phosphomolybdic acid solution and heat to develop color. It was.
TLC analysis confirmed the formation of sphingomyelin and sphingosylphosphorylcholine.

<Results of TLC analysis>
In the control (without inhibitor), sphingomyelin was deacylated with SCDase and converted to sphingosylphosphorylcholine.
In N-tosyl-1-octanesulfonylphytosphingosine (Compound 1), as the concentration increased, the production of sphingosylphosphorylcholine decreased and an inhibitory activity was observed.
In addition, N-tosyl-1-ethanesulfonyl phytosphingosine (compound 2) and compounds 5 to 9 were also found to have inhibitory activity due to decreased production of sphingosylphosphorylcholine.
On the other hand, N-tosylphytosphingosine, similar to the control, produced sphingosylphosphorylcholine, indicating that it has no inhibitory activity against SCDase.

Test example 2
<Application test in atopic dermatitis model mouse>
NC-mice, which is a model mouse for atopic dermatitis, and N-tosyl-1-octanesulfonylphytosphingosine [5% (w / v), solvent (ethanol: acetone = 1: 1 (v / v))], the solvent alone was applied to the control group at 0.2 mL at a time, twice a day every 8 hours.
It was applied for 1 week, and the number of scratches on the application area per hour was counted on the 1st day, 1st day, 5th day, and 7th day. The result is shown in FIG.

Since the phytosphingosine derivative of the present invention has an inhibitory action on an enzyme (SCDase) known to cause deacylation of sphingomyelin, it is presumed to exhibit the inhibitory activity of sphingomyelin deacylase. The
Moreover, since the phytosphingosine derivative of the present invention significantly suppresses scratching behavior in atopic dermatitis model mice, it is useful as a therapeutic agent for atopic dermatitis.

Claims (6)

The following general formula [1]

“Wherein R ¹ represents a hydrogen atom or an alkyl group; R ² represents an alkyl group, R ³ and R ⁴ are the same or different and represent a hydrogen atom or an acyl group, and R ⁵ represents an alkyl group, respectively. To do. "
A phytosphingosine derivative represented by: The phytosphingosine derivative according to claim 1, wherein R ¹ is an alkyl group. The phytosphingosine derivative according to claim 1 or 2, wherein R ⁵ is a linear alkyl group having 7 to 15 carbon atoms. The following general formula [1]

“Wherein R ¹ represents a hydrogen atom or an alkyl group; R ² represents an alkyl group, R ³ and R ⁴ are the same or different and represent a hydrogen atom or an acyl group, and R ⁵ represents an alkyl group, respectively. To do. "
The therapeutic agent of atopic dermatitis which uses the phytosphingosine derivative represented by this as an active ingredient. The therapeutic agent for atopic dermatitis according to claim 4, wherein R ^{1 of the} phytosphingosine derivative is an alkyl group. The therapeutic agent for atopic dermatitis according to claim 4 or 5, wherein R ^{5 of the} phytosphingosine derivative is a linear alkyl group having 7 to 15 carbon atoms.

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