JPH09227372A - Agent for inhibiting adhesion or infiltration of leukocyte and agent for preventing or treating inflammation - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a medicine having an action on the inhibition of adhesion and infiltration of leukocyte and effective for preventing and treating inflammation, especially progressive chronic inflammation. SOLUTION: This medicine contains a samaderin derivative of the formula [R<1> is H, OH; R<2> is O, etc.; R<3> is H, O; R<4> is H, OH; R<5> is CH3 , etc.; R<6> is OH; R<7> is H or OH; R<8> , R<9> are each a group for together forming a group of the formula: -C(R<10> )H-COO-; or R<6> , R<7> are each a group for together forming a group of the formula: -O-CO-; R<8> is H; R<9> is O; R<10> is H, OH, etc.; the dotted line is a single bond or a double bond when R<2> is O, R<5> is CH3 ; when R<7> is OH and when R<8> and R<9> are each a group for together forming a group of the formula: -C(OH)H-COO-, R<4> is OH] as an active ingredient. The compound of the formula can be obtained by isolating a plant belonging to the family Simaroubaceae and subsequently purifying the isolated compound. The compound of the formula is orally administered at a daily dose of 0.1-200mg/adult by one to several portions or intravenously administered at a daily dose of 0.1-100 mg/adult by one to several portions.

Description

Detailed Description of the Invention

[0001]

The present invention relates to a compound represented by the following general formula (I):
The present invention relates to a leukocyte adhesion or invasion inhibitor, which contains a samadelin derivative represented by the formula (1) as an active ingredient, and a prophylactic or therapeutic agent for inflammation, and particularly to a prophylactic or therapeutic agent useful for progressive chronic inflammation.

[0002]

BACKGROUND OF THE INVENTION The Samaderin derivative represented by the general formula (I) (hereinafter referred to as Samaderin derivative (I)) is a Quassia indi
It is known to exist in ca and the like and have antimalarial activity and antitumor activity.

As drugs that have been clinically used as anti-inflammatory agents, steroids and non-steroidal anti-inflammatory agents for acute and chronic inflammatory diseases, chronic progressive inflammatory diseases (for example, rheumatism, degenerative diseases). (For arthritis, etc.), immunosuppressants and gold preparations are included. The mechanism of action of non-steroidal anti-inflammatory drugs is mainly suppression of inflammatory mediators. That is, it is a symptomatic action and is highly effective against acute diseases, but not so effective against chronic inflammatory diseases. Steroidal anti-inflammatory agents have very strong effects on acute and chronic inflammatory diseases, but at the same time, they have been reported to cause serious side effects, and therefore caution should be exercised when using them. The gold preparation is not applied to acute inflammatory diseases and is used for chronic rheumatism and the like, and since it has an immunomodulatory effect, its effect is delayed. However, gold preparations have also been reported to have side effects such as mucocutaneous symptoms, bone marrow suppression, renal damage, and respiratory damage. Therefore, as with steroidal anti-inflammatory drugs, sufficient caution is required in their use. Some immunosuppressive agents have been attracting attention for clinical application to chronic rheumatism, but side effects peculiar to the immunosuppressive agents are concerned.

It is known that leukocytes have enhanced adhesion and infiltration to vascular endothelial cells at inflammatory sites. Therefore, it is considered that inflammation can be suppressed by suppressing the adhesion of leukocytes to the blood vessel wall and the subsequent infiltration outside the blood vessel. In fact, in an animal model of acute or chronic inflammation,
It has been reported that inflammation can be suppressed by specifically suppressing the accumulation of leukocytes in the inflamed area [Inflammation, 12
(4), 313-317 (1992), The Journal of Immunology, 1
47, 4167-4171 (1991), Cellular Immunology, 142, 32.
6-337 (1992)]. Therefore, we will develop a drug with a new mechanism of action, which is different from the conventional ones, that is, a drug with a function of regulating the expression and expression of adhesion molecules that play an important role in adhesion and infiltration of leukocytes, and especially in adhesion. By doing so, it is considered that problems such as conventional side effects can be avoided.

An object of the present invention is the Samadelin derivative (I).
To provide a new pharmaceutical use of

[0006]

Means for Solving the Problems As a result of intensive studies, the present inventors have found that samadelin derivative (I) has an action of suppressing leukocyte adhesion and infiltration, and is useful as an agent for preventing or treating inflammation. It was found that the present invention has been completed.

That is, the present invention relates to a leukocyte adhesion or invasion inhibitor, and a prophylactic or therapeutic agent for inflammation, which contains the samadelin derivative (I) represented by the general formula described below as an active ingredient.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION The samadelin derivative (I) used in the present invention has the general formula (I):

[0009]

Embedded image

[In the formula, R ¹ represents a hydrogen atom or —OH,
R ² is ═O, —OH or

[0011]

Embedded image

R ³ is a hydrogen atom or ═O, R ⁴ is a hydrogen atom or —OH, and R ⁵ is —CH ₃ or —CO.
Indicates OCH ₃ . ^{R 6, R 7, R 8} , R 9 is, R ⁶ is -
OH, R ⁷ represents a hydrogen atom or —OH, R ⁸
And R ⁹ are both

[0013]

Embedded image

(In the formula, R ¹⁰ is a hydrogen atom, —OH or —
Or showing a O-CO-CH indicates a ₃₎ group forming, or represents a group R ⁶ and R ⁷ form a -O-CO- together, R ⁸ represents a hydrogen atom, R ⁹ is = O Indicates.

[0015]

Embedded image

Represents a single bond or a double bond, one in the general formula represents a single bond and the other represents a double bond. Where R ²
If There is = O, R ⁵ represents a -CH _3. R ⁷ is -O
H and R ⁸ and R ⁹ are both

[0017]

Embedded image

When it is a group forming R, R ⁴ represents —OH. The Samaderin derivative (I) is, for example, the 38th Annual Conference on Fragrances / Terpenes and Essential Oil Chemistry (1994.10.8-10) Proceedings (p.329-3).
30) according to the method described in 30).
It can be isolated and purified from sia indica. In particular,

[0019]

Embedded image

And the like.

[0021]

[Operations and effects] Samadelin derivative (I), which is an active ingredient of the present invention, has an effect of suppressing leukocyte adhesion and infiltration, and is useful for treatment and prevention of various pathological conditions such as inflammation and suppression of lesion progression. It is useful. Furthermore, since adhesion molecules that play an important role in adhesion of leukocytes are not only involved in inflammation but also involved in immunosuppression and arteriosclerosis, application to these diseases is expected.

When the samadelin derivative (I) is used as the above-mentioned drug, a pharmaceutically acceptable additive (for example,
(Carriers, excipients, diluents, etc.) are appropriately mixed with pharmaceutically necessary components to prepare a pharmaceutical composition in the form of powder, granules, tablets, capsules, syrups, injections, etc. It can be administered orally.

In the above-mentioned preparation, the Samaderin derivative (I) is included.
An effective amount of The dose varies depending on the compound used, the route of administration, the symptoms, the body weight or age of the patient, etc., but when administered orally to an adult patient for the treatment of inflammation, 0.1 to 200 mg / human / day, particularly 0.5 to 100 mg
/ Human / day is preferably divided into 1 to several times per day. In the case of intravenous administration, 0.1-100 mg / human /
It is desirable to administer daily, particularly 0.5 to 50 mg / human / day, divided into 1 to several times a day.

[0024]

EXAMPLES In order to explain the present invention in more detail, experimental examples and formulation examples will be given, but the present invention is not limited thereto.

Experimental Example 1: Neutrophil-Vascular Endothelial Cell Adhesion Inhibition Test Human umbilical vein-derived vascular endothelial cells (hereinafter abbreviated as HUVEC) were placed in 96-well plates in 100 μl culture medium (EGM-UV) per well. The cells were cultured at 37 ° C. under 5% CO ₂ until they reached confluence (about 25,000 cells / cm ² ). Then, once the culture medium (EGM-UV) was removed, tumor necrosis factor (TNF-α) 5JRU (Japanese R
efference Unit) / ml and the culture medium (EGM-U
V) 100 μl was added, and the mixture was allowed to act for 4 hours at 37 ° C. under 5% CO ₂ . On the other hand, BCECF-AM [2,7
-Bis- (2-carboxyethyl) -5,6-carboxyfluorescein acetoxymethyl ester] labeled human neutrophils (1.0 x 10 ⁶ cells / ml HBSS
(Prepared into Hanks-Balance-Salt-Solution)), the drug was allowed to act for 30 minutes. 100 μl of this human neutrophil suspension (1.0 × 10 ⁶ cells / ml) was added to TNF-
The α and drug were added to HUVEC, which was allowed to adhere for 30 minutes, and non-adherent cells were removed by centrifugation at 50 × g. The cells were treated with Triton X and measured at an excitation wavelength of 508 nm and a fluorescence wavelength of 531 nm to measure the number of adhered neutrophils. The adhesion inhibition rate was calculated by the following formula. Adhesion inhibition rate (%) = 100 × (T−S) / (T−C) (In the formula, C is the number of adherent neutrophils without treatment, and T is TNF-α.
Adhesion neutrophil number at the time of action, S shows the adhesion neutrophil number at the time of acting a drug and TNF-α) FIGS.
The relationship between MA-63, samaderin C, indakacin C and indakacin X concentrations and the neutrophil adhesion inhibition rate is shown. Adhesion-inhibiting activity was observed for all of samadelin B, samadelin X, MA-63, samaderin C, indakacin C and indakacin X.

Experimental Example 2: Effect on carrageenin pleurisy [lambda] -carrageenan (Sigma) was dissolved in physiological saline,
It was adjusted to 0.25%. The physiological saline was used for intravenous injection.
Samaderin B and Samaderin X were administered at 1 mg / kg. SD (Sprague-Dawley) male rats (6 weeks old) were lightly anesthetized with ether, and 0.25%
A carrageenin solution was injected into the right thoracic cavity. At the same time, each drug was administered via the tail vein in the amounts shown above. One hour later, the same amount of drug was administered through the tail vein. After 3 hours, the rat was again anesthetized with ether, and the abdomen was opened, and then the blood was killed from the abdominal aorta. After the thoracotomy, the exudate accumulated in the chest cavity was collected and the amount of the exudate was measured. After the collected exudate was diluted with physiological saline, the number of infiltrating leukocytes was measured using a hemocytometer. FIG. 7 shows the relationship between the drug used, the amount of exudate and the number of infiltrating leukocytes. From this result, the samadelin derivative was compared to the control group administered with physiological saline,
It can be seen that the amount of exudate and the number of infiltrating leukocytes are small, and inflammation is strongly suppressed.

Formulation Example 1 (tablets) In one tablet (1) Samadelin derivative (I) 30 mg (2) Lactose 39 mg (3) Microcrystalline cellulose 24 mg (4) Carmellose calcium 4 mg (5) Hydroxypropyl cellulose 2 mg (6) Magnesium stearate 1 mg Total amount 100 mg (1) to (4) were mixed uniformly, to which the aqueous solution (5) was added, kneaded, dried and sized, ( After 6) was added and mixed, the mixture was compression-molded to produce 100 mg tablets.

Formulation Example 2 (granule or fine granule) In 1 g of (1) samadelin derivative (I) 30 mg (2) lactose 850 mg (3) low-substituted hydroxypropyl cellulose 105 mg (4) hydroxypropyl cellulose 10 mg (5) Light anhydrous silicic acid 5 mg (1) to (3) are mixed uniformly, the aqueous solution of (4) is added and kneaded, and then granulated by an extrusion granulator,
After drying and sizing, (5) was added and mixed to prepare granules. In addition, a fine granule can be prepared by omitting the extrusion granulation step in this formulation.

Formulation Example 3 (hard capsule) In one capsule (1) Samadelin derivative (I) 30 mg (2) Lactose 27 mg (3) Microcrystalline cellulose 10 mg (4) Corn starch 10 mg (5) Hydroxypropyl Cellulose 2 mg (6) Magnesium stearate 1 mg Total amount 80 mg (1) to (4) were uniformly mixed, to which the aqueous solution (5) was added, kneaded, dried and sized, After adding (6) and mixing, 80 mg of this product was filled in a hard capsule to prepare a hard capsule.

Formulation Example 4 (Syrup) In 100 ml (1) Samadelin derivative (I) 0.2 g (2) D-sorbitol solution (70%) 70 g (3) Citric acid 0.3 g (4) Paraoxybenzoic acid Methyl 0.028 g (5) Propyl paraoxybenzoate 0.012 g (6) Propylene glycol 10 g (7) Purified water A suitable amount of (1), (4) and (5) is dissolved in (6), and further (2) And (3) were added and stirred until uniform, then (7) was added to bring the total amount to 1
The amount was set to 00 ml and used as a syrup.

Formulation Example 5 (Injection) In 2 ml (1) Samadelin derivative (I) 10 mg (2) Hydrochloric acid 0.1 ml (3) Sodium hydroxide proper amount (4) Distilled water for injection proper amount A mixture solution of the above components After that, an injection was prepared by a conventional method.

[Brief description of drawings]

FIG. 1 is a graph showing the relationship between samadelin B concentration and neutrophil adhesion inhibition rate.

FIG. 2 is a graph showing the relationship between the concentration of samadelin X and the inhibition rate of neutrophil adhesion.

FIG. 3 is a graph showing the relationship between MA-63 concentration and neutrophil adhesion inhibition rate.

FIG. 4 is a graph showing the relationship between samadelin C concentration and neutrophil adhesion inhibition rate.

FIG. 5 is a graph showing the relationship between indakacin C concentration and neutrophil adhesion inhibition rate.

FIG. 6 is a graph showing the relationship between indakacin X concentration and neutrophil adhesion inhibition rate.

FIG. 7 shows the effect of samadelin B and samadelin X on rat carrageenin pleurisy model.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Harumasa Toya 6-6 Naka-cho 6-6, Koyo-cho, Higashinada-ku, Kobe-shi, Hyogo Prefecture Room 404, Room 611 (72) Inventor Satoshi Nakata 2-10-27 Onohara Nishi, Minoh City, Osaka Prefecture Rakuzanso 2-E (72) Inventor Masumi Furukawa 9-29 Higashisanodai, Izumisano, Osaka (72) Inventor Takeshi Masagaki 45-B-903 Yamadaminami, Suita, Osaka

Claims (4)

[Claims] 1. A compound of the general formula (I): [In the formula, R ¹ represents a hydrogen atom or —OH, R ² represents ═O,
-OH or , R ³ represents a hydrogen atom or ═O, R ⁴ represents a hydrogen atom or —OH, and R ⁵ represents —CH ₃ or —COOCH ₃ . ^{R 6, R 7, R 8} , R 9 is, R ⁶ represents -OH,
R ⁷ represents a hydrogen atom or —OH, and R ⁸ and R ⁹ are both (In the formula, R ¹⁰ is a hydrogen atom, —OH or —O—CO—C.
Or a group which forms a shows the H _3), or represents a group R ⁶ and R ⁷ form a -O-CO- together, R ⁸ represents a hydrogen atom, a R ⁹ is = O. Embedded image Represents a single bond or a double bond, and in the general formula, one represents a single bond and the other represents a double bond. Here, when R ² = is O, R ⁵ represents a -CH _3. R ⁷ is —OH, and R ⁸ and R ⁹ are both Is a group which forms a, R ⁴ is bonded or invasion inhibitor of leukocytes to Samaderin derivative as an active ingredient represented by showing the -OH]. 2. A samadelin derivative is represented by: The leukocyte adhesion or infiltration inhibitor according to claim 1, which is one selected from the group consisting of: 3. General formula (I): [In the formula, R ¹ represents a hydrogen atom or —OH, R ² represents ═O,
-OH or , R ³ represents a hydrogen atom or ═O, R ⁴ represents a hydrogen atom or —OH, and R ⁵ represents —CH ₃ or —COOCH ₃ . ^{R 6, R 7, R 8} , R 9 is, R ⁶ represents -OH,
R ⁷ represents a hydrogen atom or —OH, and R ⁸ and R ⁹ are both (In the formula, R ¹⁰ is a hydrogen atom, —OH or —O—CO—C.
Or a group which forms a shows the H _3), or represents a group R ⁶ and R ⁷ form a -O-CO- together, R ⁸ represents a hydrogen atom, a R ⁹ is = O. Embedded image Represents a single bond or a double bond, and in the general formula, one represents a single bond and the other represents a double bond. Here, when R ² = is O, R ⁵ represents a -CH _3. R ⁷ is —OH, and R ⁸ and R ⁹ are both And R ⁴ represents —OH, the prophylactic or therapeutic agent for inflammation comprising a samadelin derivative represented by the formula: 4. A samadelin derivative is represented by: The preventive or therapeutic agent for inflammation according to claim 3, which is one selected from the group consisting of: