JP2513452B2 - Histamine release inhibitor - Google Patents

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a histamine release inhibitor containing a sembrane type diterpene compound as an active ingredient.

[0002]

2. Description of the Related Art Sembran type diterpenes are goat eyes ( G
orgonacea ) and the order of Umitosaka ( Alcyo )
nuclea ), which has recently been found to have antitumor activity and has attracted attention [for example, B. Tursch et al., Tetrahedron 31: 129, 19].
1975, England and Weinheimer (AJW
Einheimer et al., Tetrahedron Letters 2923
Page, 1977, United Kingdom].

In view of such circumstances, Hirayama et al., Sarcoph, a coelenterate animal belonging to the order of the sea turtle, Sarcoph
As a result of intensive investigations focusing on yton glaucum ), a novel sembrane-type diterpene compound having an anti-solid tumor action was found [eg, JP-A-56-61].
No. 318]. However, it has only been reported so far about the anti-solid tumor action of the compound, and there has been no report on the anti-inflammatory action and the action on inflammatory cells (polymorphonuclear leukocytes / mast cells).

Mast cells, which are a type of inflammatory cells, are basophilic granular cells that are metachromatically positive for basic pigments widely distributed in connective tissue, and release histamine and leukotriene, which are mediators of allergic reaction. It has been known. From the era when connective tissue was understood only from its function as a supporting tissue of the living body, at present, the supply of cell nutrient, excretion of cell metabolism, preservation of cell environment, and the group of cells against extrinsic invasion by becoming a place of inflammation It has changed so that it can be caught as a place to control various cellular activities such as protection. Mast cells themselves have been re-recognized as important cells that carry various physiological activities due to recent advances in cell biology and biochemistry (Metabolism, Vol. 13, No. 5, Special Issue on Mast Cells, Nakayama Shoten).

[0005] For example, in recent years, new findings have been accumulated on the etiology of ulcerative colitis, and the persistent inflammation of the large intestine mucosa, which is the main symptom of this disease, is manifested in the mechanism of defense against invasion of antigens such as bacteria from the intestinal tract. It is believed that the foundation is the breakdown and establishment of autoimmunity to the large intestine. Both immediate and delayed allergies are involved in this disease. In the acute stage, establishment of immediate-type allergy in the large intestine mucosa is in the foreground of the pathological condition. That is, at this time, high plasma histamine levels, large intestinal mucosal eosinophilia, decreased mast cells, etc. are observed. In response to this immediate allergy, induction of vasoactive substances, microcirculatory disturbance of the intestinal tract occurs,
Symptoms in the acute phase appear due to increased permeability of the intestinal mucosa and enhanced muscle contraction. It has already been reported that cromoglycate, which is a histamine release inhibitor that suppresses mast cell degranulation, is effective in ulcerative colitis. That is, it is expected that a compound showing a strong inhibitory action on histamine release from mast cells will be very effective in treating ulcerative colitis.

It has recently been known that histamine plays an important role not only as a mediator in the early stage of acute inflammation but also as a regulator of chronic inflammation.
For example, there is a receptor for histamine in fibroblasts, and it is said that histamine promotes collagen production. It has also been confirmed that mast cells increase in the fibrous tissue. Therefore, it is considered that suppressing the release of histamine by acting on mast cells indirectly suppresses the promotion of fibrosis, that is, the promotion of collagen production in the tissue in which inflammation is becoming chronic.
For example, it is useful for suppressing diseases associated with abnormal growth of fibers, pulmonary fibrosis, post-treatment keloid treatment, and abnormal growth of fibers due to trauma.

[0007]

DISCLOSURE OF THE INVENTION The present inventors have been carrying out research aiming to develop an effective ingredient having an action of suppressing histamine release,
Focusing on the effect on the skin of the sembrane-type diterpene compound contained in the coelenterate C. elegans, as a result of intensive studies, it was found that the compound has a strong inhibitory effect on histamine release, and the present invention was completed. Came to do.

[0008]

That is, the present invention provides the following general formula (I):

[0009]

Embedded image

A histamine release inhibitor containing a sembrane type diterpene compound represented by the formula (wherein R represents a hydrogen atom or an acyl group) as an active ingredient. Hereinafter, the present invention will be described in detail.

The cembrane-type diterpene compound represented by the above general formula (I) used in the present invention is a coelenterate animal belonging to the order of the sea turtle, Sarcophyton.
glaucum ), for example, JP-A-56-6131
It is obtained by the method described in JP-A-8. That is, the compound according to the present invention can be separated by silica gel column chromatography from the lipid fraction of the extract of C. communis.

[0012] Greater mushrooms usually live in the coral reefs of the Indian Ocean and the Pacific Ocean, for example, the Greater Mushrooms that inhabit the Red Sea are known to contain sarcophines and 16-deoxo sarcophines [Bernstein (J. Bernstein, et al., Tetrahedron, Vol. 30, page 2817, 1974, England and Y. Kashman, et al., Tetrahedron, Vol. 30, page 3615, 1974, England]. The components contained in C. terrestris mushrooms may differ depending on the time of collection and the place of collection, so it is necessary to appropriately select and decide.

During extraction, it is preferable to dehydrate and shred so that the surface becomes less viscous. Extraction solvents include, for example, alcohols such as methanol, ethanol, isopropanol; halogenated hydrocarbons such as chloroform; hydrocarbons such as benzene, hexane, heptane; ethers such as ethyl ether, isopropyl ether, dioxane; acetone, methyl ethyl ketone. And the like; organic solvents such as esters such as ethyl acetate, and mixed solvents thereof.

During the extraction operation, in order to avoid decomposition of the contained components, it is preferable to make the contact area with air as small as possible, or to carry out in an inert gas atmosphere. The extraction can be carried out at room temperature, but in order to accelerate the extraction, it may be carried out under heating. The solvent of the extract obtained by separating from the residue by a conventional method can be distilled off by a conventional method to obtain a crude extract.

The crude extract may be treated with activated carbon or a conventional method [for example, J. Folch's method--Fauls et al., Journal of Biological Chemistry (J.
Biol. Chem. ) 226, 497, 1957
Year, refer to US country]. The lipid fraction of the extract of C. mellifera thus obtained is a viscous brown oil.

For further purification, chromatography may be used. Chromatography may be either column chromatography or preparative thin layer chromatography. As a packing material for column chromatography,
Silica gel, alumina, cellulose powder, activated carbon and the like are used. The elution solvent may be appropriately selected and determined according to the filler, but when silica gel is used as the filler, hexane, hexane-ethyl acetate (volume ratio 0.95 to 0.9: 0.05 to 0.1) and the like are preferable. Further, the crude fraction obtained here is further purified by column chromatography by changing the packing material and the elution solvent.
It can also be isolated.

As the gel for preparative thin layer chromatography, silica gel, alumina, cellulose powder or the like is used. As the developing solvent, a hexane-acetic acid ethyl ester mixed solvent is suitable. Among the compounds represented by the general formula (I) thus obtained, those in which R is a hydrogen atom are represented by Sarcophitol (Sarcophitol).
ytol) -A. When this sarcophytol-A is acylated by a conventional method, sarcophytol-
When an acylated product such as A acetate is further subjected to ester decomposition of this sarcophytol-A acylated product by a conventional method, sarcophytol-A is obtained.

As the active ingredient of the present invention, it is not always necessary to use a pure product of the sembrane-type diralpen compound represented by the general formula (I), which is isolated as described above, and a mixture of these substances or a mixture thereof may be used. It is also possible to use the extract, partially purified product and the like. The cembran-type diterpene compound obtained by the above-mentioned method or an extract or partially purified product of C. mellifera containing the compound is added to the drug of the present invention as an active ingredient.

The histamine release inhibitor according to the present invention can be administered by any method, but preferably the following method is carried out. That is, parenteral administration such as subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection and the like, as well as oral administration are possible.

The dose is determined according to the age, health condition, weight of the patient, type of treatment, if any, and frequency of treatment, nature of desired effect, etc., but generally the daily dose of active ingredient Is 50 to 2,000 mg, especially 100 to 500 mg
Is preferable, and it is administered once or dividedly (preferably orally). In the case of oral administration, it is used in the form of tablets, capsules, powders, elixirs, etc., and in the case of parenteral administration, it is used in the form of liquid such as liquid or suspension and sterilized liquid form. When used in the form as described above, a solid or liquid non-toxic pharmaceutical carrier may be included in the composition.

As an example of the solid carrier, a usual gelatin type capsule is used. In addition, the active ingredient is tableted or powdered with or without an auxiliary drug. These capsules, tablets and powders generally contain 5-95% by weight, preferably 25-90% by weight of the active ingredient. As the liquid carrier, animal or plant origin oil such as peanut oil, soybean oil, mineral oil, sesame oil, or synthetic oil can be used. In general, saline, dextrose or similar saccharide solution, glycols such as ethylene glycol, propylene glycol, polyethylene glycol are preferred as liquid carriers.

When administered parenterally by intramuscular injection, intravenous injection or subcutaneous injection, it is used as a sterile solution to which other solutes such as salt or glucose are added in order to make the solution isotonic. Suitable solvents for injection include sterile water, lidocaine hydrochloride solution (for intramuscular injection), physiological saline, glucose, liquid for intravenous injection, electrolyte solution (for intravenous injection)
Etc. In the case of these injections, it is usually 0.
The active ingredient may be contained in an amount of 5 to 20% by weight, preferably 1 to 10% by weight. For orally administered liquids,
Suspensions or syrups containing 0.5-10% by weight of active ingredient are preferred. As the carrier in this case, fragrance, syrup,
An aqueous excipient such as a pharmaceutical micelle is used.

[0023]

EXAMPLES The present invention will be described in more detail with reference to examples below, but the present invention is not limited to the following examples unless it exceeds the gist thereof. Example 1 1% (W / V) lambda carrageenin (λ-carrag) was added to the left foot pad of Wistar male rats (120 to 150 g).
eenin) 0.75 ml subcutaneously injected, left foot 3
Causes edema peaking after ~ 4 hours (rat carrageenan paw edema method). It is known that histamine is involved in this edema immediately after the injection of carrageenin, followed by prostaglandins, leukotrienes, and further chemical mediators such as bradykinin, which is an excellent model of inflammation.

Using this model, sarcophytol A was given orally in advance, and 1 hour after that, carrageenin was injected to induce inflammation and the effect on edema was examined (edema was measured by a foot volume meter). The edema rate was calculated as the rate of increase in foot volume after inflammation was induced relative to the foot volume before inflammation was induced. That is,

[0025]

[Equation 1] Table 1 shows the results.

[0026]

[Table 1]

Example 2 In vitro using mast cells prepared from rat peritoneal cavity
It is possible to reproduce histamine release from mast cells in tro. That is, mast cells are pre-incubated with an IgE antibody (for example, an IgE antibody prepared by sensitizing a rabbit with a protein prepared from the epidermis of Ascaris suum), and then IgE
Addition of an antigen (eg, a protein prepared from Ascaris epidermis) results in histamine release from mast cells. Such release of histamine is caused by artificial stimulant compound 48/80, A
It can also be produced by adding TP (adenosine triphosphate) to mast cells. When the compound 48/80 and a protein extracted from the epidermis of Ascaris suum were used as stimulants to examine the action in vitro, it was confirmed that the substance has a strong histamine release inhibitory action.

That is, mast cells were collected from the abdominal cavity of male Wistar rats weighing 400 to 450 g according to a conventional method. 5 × 10 ⁴ cells were added to Krebs Ringer buffer (pH
Compound 48/80 after 10 minutes incubation in 7.4)
Was added to 100 ng. After further culturing for 10 minutes, cells were removed by centrifugation (3000 rpm × 5 minutes), and histamine in the supernatant was removed by the method of Shore et al. (J. Phar. Ex.
p. Ther, Vol 127 , 182-186 (195).
9)). The total amount of histamine contained in mast cells was separately determined, compound 48/80 was treated as a stimulant, and the amount of histamine released thereafter was determined as a ratio to the total amount of histamine. That is

[0029]

[Equation 2]

The results are shown in Table 2.

[Table 2]

[0031]

INDUSTRIAL APPLICABILITY The histamine release inhibitor of the present invention acts on mast cells and strongly suppresses histamine release, and is therefore expected to be effective against various diseases caused by histamine. . That is, it is not only useful as an antiallergic agent, but is also effective against diseases in which the pharmacological mechanism has been recently analyzed and it is speculated that mast cells are involved in its etiology, such as ulcerative colitis. .

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication C07C 69/145 C07C 69/145

Claims (1)

(57) [Claims] 1. A compound represented by the following general formula (I) A histamine release inhibitor comprising a sembrane-type diterpene compound represented by the formula (wherein R represents a hydrogen atom or an acyl group) as an active ingredient.