KR20000026054A - Fructigenine derivatives, pharmaceutically acceptable salt thereof and anti-inflammatory composition containing them - Google Patents

Abstract

PURPOSE: Fructigenine A derivatives, a pharmaceutically acceptable salt thereof and an anti-inflammatory composite ion containing the same are provided which can be used for treating inflammation effectively. CONSTITUTION: Fructigenine A derivatives of formula(1) as indole alkaloid prepared from strains of Penicillium sp. CJ-73 producing an anti-inflammatory substance and a pharmaceutically acceptable salt thereof and an anti-inflammatory composite ion containing the same useful as a medicament for the treatment of inflammation are described. In formula, R1 is C2-C4 alkenyl or C1-C4 alkyl, R2 is H, substituted or non substituted C1-C4 alkyl, -COCH2R6(R6 is H, C1-C6 alkyl, cyclo C3-C6 alkyl, heteroarylalkyl, substituted or non substituted phenyl or heteroaryl) or -COCHR7R8(R7R8 are the same or different H, C1-C4 alkyl, cyclo C3-C6 alkyl, heteroarylalkyl, substituted or non substituted phenyl or heteroaryl), R3 is H or C1-C4 alkyl, R4 and R5 are the same or different H or substituted or non substituted C1-C4 alkyl, aryl or benzyl. The compound has one and a half times or more the anti-inflammatory effect of indomathacin as a conventional anti-inflammatory agent by dropsy experimentation on mice.

Description

Derivatives of fructogenin A and pharmaceutically acceptable salts thereof and anti-inflammatory compositions containing them

The present invention relates to the synthesis of derivatives of alkaloids with anti-inflammatory efficacy. More specifically, the present invention relates to derivatives of fructogenin A of formula (1), which are alkaloid derivatives produced in a strain of the genus Penicillium , and pharmaceutically acceptable salts thereof and anti-inflammatory compositions comprising them.

<Formula 1>

Wherein R ¹ represents C ₂ -C ₄ alkenyl or C ₁ -C ₄ alkyl,

R ² is a hydrogen atom or substituted or unsubstituted C ₁ -C ₄ alkyl, wherein —COCH ₂ R ⁶ (wherein R ⁶ is a hydrogen atom, C ₁ -C ₄ alkyl, cycloC ₃ -C ₆ alkyl, heteroaryl A radical of alkyl, substituted or unsubstituted phenyl or heteroallyl, or a formula -COCHR ⁷ R ⁸ , wherein R ⁷ and R ⁸ are the same or different and represent a hydrogen atom, C ₁ -C ₄ alkyl, cycloC ₃ -C ₆ alkyl, heteroarylalkyl, substituted or unsubstituted phenyl or heteroallyl, or when R ⁷ is hydrogen, R ⁸ is a halogen atom selected from fluoro, chlorine, bromine and iodine or R ⁶ and R ⁷ together may form a cycloallyl or cycloalkyl group)

R ³ is a hydrogen atom or C ₁ -C ₄ alkyl,

R ⁴ and R ^{5, which} are the same or different, represent a hydrogen atom or represent a substituted or unsubstituted C ₁ -C ₄ alkyl, aryl or benzyl group.

Fructigenine A is an indole alkaloid found in Penicillum fructigenium by Kunizo et al in Japan in 1989 and has a molecular weight of 443.5 and a molecular formula of C ₂₇ H. ₂₉ N ₃ O ₃ and represented by the following chemical formula (2) (Chem. Pharm. Bull 37 (11) 2937-2939, 1989).

Kunizo et al. First identified the structure of the above compounds in the above paper and reported that they had an effect of inhibiting the growth of plant stem ends, but there was no mention of anti-inflammatory effects. In addition, no pharmacological effects on the anti-inflammatory effects of fructogenin A have been reported.

It is therefore an object of the present invention to provide novel derivatives of fructogenin A and pharmaceutically acceptable salts thereof and anti-inflammatory compositions comprising them.

The present inventors, while searching for a substance having an anti-inflammatory action in the fermentation product of the microorganism, found that the indole alkaloid compound of the formula (1) produced by the fungus belonging to the genus Penicillum has a strong anti-inflammatory effect The present invention has been completed by confirming that derivatives containing fructogenin A exhibit potent anti-inflammatory activity.

Thus, according to the present invention, there is provided a compound of formula (1) and a pharmaceutically acceptable salt thereof and an anti-inflammatory composition comprising them.

<Formula 1>

Wherein R ¹ represents C ₂ -C ₄ alkenyl or C ₁ -C ₄ alkyl,

R ² is a hydrogen atom or substituted or unsubstituted C ₁ -C ₄ alkyl, wherein —COCH ₂ R ⁶ (wherein R ⁶ is a hydrogen atom, C ₁ -C ₄ alkyl, cycloC ₃ -C ₆ alkyl, heteroaryl A radical of alkyl, substituted or unsubstituted phenyl or heteroallyl, or a formula -COCHR ⁷ R ⁸ , wherein R ⁷ and R ⁸ are the same or different and represent a hydrogen atom, C ₁ -C ₄ alkyl, cycloC ₃ -C ₆ alkyl, heteroarylalkyl, substituted or unsubstituted phenyl or heteroallyl, or when R ⁷ is hydrogen, R ⁸ is a halogen atom selected from fluoro, chlorine, bromine and iodine or R ⁶ and R ⁷ together may form a cycloallyl or cycloalkyl group)

R ³ is a hydrogen atom or C ₁ -C ₄ alkyl,

R ⁴ and R ⁵ are the same or different and represent a hydrogen atom or a substituted or unsubstituted C ₁ -C ₄ alkyl, aryl or benzyl group.

Hereinafter, the present invention will be described in more detail.

As used herein, the term “C ₁ -C ₄ alkyl” refers to a saturated straight or branched chain hydrocarbon radical having 1 to 4 carbon atoms such as methyl, ethyl, isopropyl, n-butyl, isobutyl, t-butyl and the like. And the term "halogen atom" means a halogen atom selected from fluoro, bromine, chlorine and iodine.

Among the compounds of the formula (1), preferred compounds are R^OneIs ethylene or ethyl and R²Are hydrogen, alkyl and allylcarbonyl derivatives and R³Is a hydrogen atom, R⁴, R⁵Is a hydrogen atom, substituted or unsubstituted C_One-C₄Compound of formula (1) which is an alkyl, aryl or benzyl group.

As described above, the novel compounds of formula (I) according to the invention may be administered to patients in need thereof in admixture with pharmaceutically acceptable excipients or carriers.

The compound can be very useful for healing inflammation. This compound can be used in a similar way to the typical anti-inflammatory drugs indomethacin, hydrocortisone and anti-inflammatory drugs.

Typically, a daily dose level of the compound of formula (1) is effective to administer about 5 to 10 mg of active ingredient per kg of body weight when orally administered to an inflammatory mammal. In some cases, the daily dose may be administered in several divided doses, for example 2 to 4 times a day. More preferred dosages may vary depending on the particular compound to be used, weight, sex, health condition, diet, time of administration, method of administration, and the severity of the disease.

The compounds of the present invention can be administered in the form of oral, injectable, and topical preparations, although not particularly limited thereto, as desired.

Solid form preparations for oral administration include capsules, tablets, pills, powders and granules, in which the active substance is sucrose, lactose, dicalcium phosphate, cellulose, pectin, dextrin, gelatin or It is mixed with at least one of inert solid carriers such as starch. Solid form preparations may also contain additional ingredients (such as lubricants such as magnesium stearate) other than inert solid carriers, as in conventional methods, and in the case of capsules, tablets and pills may also contain buffering agents. have.

Liquid form preparations for oral administration include pharmaceutically acceptable emulsions prepared by containing adjuvants such as wetting agents, emulsifiers, suspending agents, sweetening agents, fragrances and flavorings, including inert diluents commonly used in the art, such as water, Solutions, suspensions and syrups.

Compositions for rectal administration as topical preparations include suppositories which may contain, in addition to the active substance, excipients such as cocoa butter or suppository waxes.

On the other hand, the pharmaceutical preparation contains a pharmaceutically acceptable carrier and a metal salt of the compound of formula (I). Carriers include conventional hydrophilic or oil-water carriers that are dispersible in water, in particular oil-water emulsions that are dispersible or water-soluble in conventional semi-soft or creamy water, with minimal discomfort to the infection surface. Can be applied while giving. This composition can be prepared by simply mixing or homogeneously mixing the finely divided active compound with a hydrophilic carrier or base plaster.

Fructininin A (MS-73) according to the present invention is prepared as known in the literature (Chem. Pharm. Bull 37 (11) 2937-2939, 1989). It is possible to produce various derivatives using the obtained fructogenin A. Briefly described as follows.

Dihydrofruptogenin A is obtained by dissolving fructogenin A in an alcohol solvent such as ethanol, followed by a hydrogenation reaction at room temperature to remove the solvent. The reaction time is usually about 1 day. N-diacetyl fructogenin A can be prepared by dissolving fructogenin A in a solvent such as methylene chloride and then carrying out a reduction reaction at -78 ° C. N-propionyl fructogenin A can be prepared by subjecting N-diacetyl fructogenin A to an acylation reaction in an organic solvent such as methylene chloride.

Physicochemical properties of the prepared compounds can be confirmed by methods well known in the art. For example, the prepared compound of formula (1) can be analyzed by HR-MS (high resolution mass spectroscopy), H-NMR, C-MNR and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited in any way by these.

<Example 1>

Preparation of Dihydrofructigenin A (R ¹ = -CH ₂ CH ₃ , R ² = -COCH ₃ , R ³ = H, R ⁴ = H, R ⁵ = CH ₂ Ph)

Fructininin A (R ¹ = -CH = CH ₂ , R ² = -COCH ₃ , R ³ = H, R ⁴ = H, R ⁵ = CH ₂ Ph, 50 mg. 0.11 mmole) and 5% Pd / C (2 mg) was dissolved in 10 mL of ethanol, and then hydrogenated at room temperature under agitation for 1 day. After the reaction was terminated, after removing the Pd / C with celite, the ethanol solution was distilled off under reduced pressure. The reaction compound was extracted with methylene chloride (40 mL) and water, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure to obtain dihydrofruptogenin A (42 mg, 83.7%).

H-NMR (CDCl ₃ ): 8.0 (1H, bs), 7.08-7.51 (8H, m), 5.91 (1H, s), 4.26 (1H, dd, J = 6.7, 2.6 Hz), 3.74 (1H, m) ), 3.51 (1H, m), 2.90 (1H, dd, J = 14.4, 4.7 Hz), 2.66 (3H, s), 2.53 (1H, dd, J = 12.4, 5.6 Hz), 2.15 (1H, t, J = 12 Hz), 1.21 (2H, q, J = 7 Hz), 0.88 (6H, d, J = 7.1 Hz), 0.83 (3H, t, J = 7.4 Hz).

<Example 2>

Preparation of N-DiacetylFrutigenin A (R ¹ = -CH = CH ₂ , R ² = H, R ³ = H, R ⁴ = H, R ⁵ = CH ₂ Ph)

Fructininin A (50 mg. 0.11 mmole) was dissolved in methylene chloride (20 mL), and lithium aluminum hydride (8.3 mg, 0.22 mmole) was slowly added dropwise at -78 ° C, followed by stirring for 4 hours. After completion of the reaction, the excess lithium aluminum hydride was consumed with a small amount of water, extracted with methylene chloride (60 mL) and water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to give N-diacetylfructinin A (32 mg , 70.7%).

H-NMR (CDCl ₃ ): 7.55-6.7 (9H, m), 6.75 (1H, t, J = 7 Hz), 6.59 (1H, d, J = 7.7. Hz), 5.98 (1H, m), 5.67 ( 1H, s), 5.13 (1H, m), 4.12 (1H, m), 3.90 (1H, m), 3.58 (1H, dd, J = 14.5, 3.7 Hz), 2.84 (1H, dd, J = 14.4, 10.3 Hz), 2.50 (1H, dd, J = 12, 6.2 Hz), 2.40 (1H, dd, J = 11.3, 10 Hz), 1.14 (3H, s), 1.00 (3H, s).

<Example 3>

Preparation of N-propionylfructigenin A (R ¹ = -CH = CH 2, R ² = COCH ₂ CH ₃ , R ³ = H, R ⁴ = H, R ⁵ = CH ₂ Ph)

N-diacetyl fructogenin A (30 mg, 0.074 mmole) is dissolved in methylene chloride (10 mL) and then triethylamine (10.3 μl, 0.074 mmole) is added dropwise. The reaction was stirred at room temperature for 30 minutes, then propionyl chloride (7 μl, 0.08 mmole) was added dropwise and stirred at room temperature for 6 hours. After completion of the reaction, the mixture was extracted with methylene chloride (30 mL), and the organic solvent layer was washed with 10% HCl solution, saturated NaHCO ₃ and water in that order, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. Propionylfruptogenin A (21 mg, 61.7%) was obtained.

H-NMR (CDCl ₃ ): 7.52-7.11 (9H, m), 6.2 (1H, s), 5.81 (1H, m), 5.6 (1H, s), 5.13 (2H, m), 4.24 (1H, m ), 3.81 (1H, m), 3.57 (1H, dd, J = 3.7, 14.4 Hz), 3.52 (1H, m), 2.93-2.78 (1H, dd, J = 10.3, 12.5 Hz), 2.64-2.53 ( 2H, m), 2.23 (1H, t, J = 12.3 Hz), 1.27 (3H, t, J = 7.2 Hz), 1.14 (3H, s), 0.88 (3H, s).

Mass (Low Resolution FAB +) = 458.2 (M + 1)

Pharmacological Example

The method of edema inhibition experiment of mice measuring anti-inflammatory efficacy was performed as follows. The experimental animals were purchased after ICR mice (succulents, 5 weeks old), and used for feeding. The feeds were fed pelleted rat feed (Cheil Jedang), and the feed and water were fed freely. Mice were maintained under conditions of 12 hours of sunshine, 22 ± 2 ° C., and 60 ± 10% humidity.

TPA (12-o-tetradecanoylphorbol-13-acetate, manufactured by Sigma) as a base salt is dissolved in acetone to 1 µg / µl and used to 10 µl / ear. The pure purified compound was dissolved in acetone to 0.6, 0.3, 0.15, 0.075 mg / ear, and the control compound indomethacin was quantitated to 0.7, 0.3, 0.15 mg / ear, and dissolved in acetone and applied locally to the ears of mice.

The thickness of the right ear of the mouse before drug treatment is measured with a caliper gauge, and the topical application drug is dissolved in acetone and instilled into the right ear by 10 μl / ear. Immediately drop TPA into the right ear at 10 μl / ear. After 5 hours of TPA treatment, the thickness of the right ear is measured.

Experimental results were measured by comparing the thickness of the ear obtained before administration with the thickness of the ear obtained after administration to determine the edema rate and edema inhibition rate.

Tt-Tn edema rate (%) = --------- × 100 TnEc-Et edema inhibition rate (%) = --------- × 100Ec Tt: Right ear thickness after TPA treatment Tn: Right ear thickness before TPA treatment Ec: Edema rate by vehicle administration Et: Edema rate by administration of test substance and positive control

The experimental results are shown in the following table. (n = 3)

Dosing medication Capacity (mg / ear) Route of administration Edema rate (%) Inhibition Rate (%) Control 0 Topical application 101.92 - MS-73 0.075 72.53 28.84 0.15 65.00 36.23 0.3 52.16 48.82 0.6 36.00 64.68 Indomethacin 0.175 88.75 12.92 0.35 61.84 39.33 0.7 44.89 56.45

The effective concentration (ED ₅₀ ) of the mouse edema test showed that ED ₅₀ of MS-73 was 0.29 mg / ear and ED ₅₀ of indomethacin was 0.68 mg / ear, indicating that MS-73 is more than twice as effective as indomethacin. It was confirmed that it has a strong anti-inflammatory effect.

Derivatives of fructogenin A were synthesized and confirmed for anti-inflammatory action in mice. The results are as follows.

Medication Capacity (mg / ear) Route of administration Edema rate (%) Edema inhibition rate (%) Control - 127.02 - Fructininin A 0.60 Topical application 26.91 78.82 Dihydro Fructininin A 0.60 29.76 76.57 N-diacetyl fructogenin A 0.60 21.33 83.21 N-propionylfructigenin A 0.60 94.07 36.64 Hydrocortisone 0.30 26.56 79.09 Indomethacin 0.60 40.05 51.56

As shown in the above table, the anti-inflammatory action on the derivatives of fructogenin A could be confirmed.

Compounds according to the present invention have been confirmed to have a strong anti-inflammatory effect showing more than half the drug efficacy compared to the known anti-inflammatory agent indomethacin in the edema experiment using a mouse, they can be effectively used for the treatment of inflammation.

Claims (3)

The compound of formula (1) and a pharmaceutically acceptable salt thereof. In the above formula, R ¹ represents C ₂ -C ₄ alkenyl or C ₁ -C ₄ alkyl, R ² is a hydrogen atom or substituted or unsubstituted C ₁ -C ₄ alkyl, wherein —COCH ₂ R ⁶ (wherein R ⁶ is a hydrogen atom, C ₁ -C ₄ alkyl, cycloC ₃ -C ₆ alkyl, heteroaryl A radical of alkyl, substituted or unsubstituted phenyl or heteroallyl, or a formula -COCHR ⁷ R ⁸ , wherein R ⁷ and R ⁸ are the same or different and represent a hydrogen atom, C ₁ -C ₄ alkyl, cycloC ₃ -C ₆ alkyl, heteroarylalkyl, substituted or unsubstituted phenyl or heteroallyl, or when R ⁷ is hydrogen, R ⁸ is a halogen atom selected from fluoro, chlorine, bromine and iodine or R ⁶ and R ⁷ together may form a cycloallyl or cycloalkyl group) R ³ is a hydrogen atom or C ₁ -C ₄ alkyl, R ⁴ and R ⁵ are the same or different and represent a hydrogen atom or a substituted or unsubstituted C ₁ -C ₄ alkyl, aryl or benzyl group. The compound of claim 1 wherein R^OneIs ethylene or ethyl and R²Are hydrogen, alkyl and allylcarbonyl derivatives and R³Is a hydrogen atom, R⁴, R⁵Is a hydrogen atom, substituted or unsubstituted C_One-C₄Compounds of formula (1) which are alkyl, aryl or benzyl groups and pharmaceutically acceptable salts thereof. An anti-inflammatory composition comprising an anti-inflammatory effective amount of a compound of formula (I) according to claim 1 and / or a salt thereof and a pharmaceutically acceptable carrier.