JPH07291857A - Preventing and therapeutic agent containing glycyrrhetinic acid compound as active ingredient - Google Patents

Abstract

PURPOSE:To obtain a preventing and therapeutic agent for bone resorption osteopathy such as malignant hypercalcemia, bone Paget disease or osteoporosis containing a glycyrrhetinic acid compound as an active ingredient. CONSTITUTION:This preventing and therapeutic agent for osteopathy contains a glycyrrhetinic acid compound expressed by the formula (R is H or glycosyl) or 3-dehydroglycyrrhetinic acid (salt) as an active ingredient. The compound is administered in a daily dose of 0.1-10mg/kg, preferably 0.3-3.0mg/kg as the sum total of the active ingredient for an adult in 1-3 divided portions.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a preventive and therapeutic agent for bone resorbable bone diseases such as malignant hypercalcemia, Paget's disease of bone and osteoporosis.

[0002]

2. Description of the Related Art In recent years, so-called geriatric diseases have increased along with a rapid increase in the elderly population. Among them, bone diseases such as osteoporosis have been desired to develop preventive and therapeutic methods as diseases that frequently cause bone fractures and lead to bedridden elderly people.

Once bone is formed, it is not a construct that does not change at all, and its structure and quantity are maintained on the balance between bone formation and bone resorption. Therefore, when the balance is lost due to aging or other causes, various bone diseases develop.

As diseases caused by abnormally increased bone resorption, malignant hypercalcemia caused by myeloma, lymphoma and the like, Paget's disease of bone caused by local bone resorption, the cause of which is unknown but aging Examples thereof include osteoporosis in which bone mass is reduced.

Bone is mainly composed of collagen fibers which are organic and calcium salts which are inorganic, and these two are combined to form a bone which is a strong structure which is strong against both tension and pressure. In particular, calcium salts account for 70% of the total bone weight, but in bone-resorbing bone diseases such as osteoporosis, calcium salts are eluted from the bones into the blood with the progress thereof, resulting in hypercalcemia. The calcium salts are gradually lost from the.

Up to now, a method for maintaining calcium has been adopted for the prevention of such diseases, and a method for supplementing calcium has been adopted for the therapy, and active vitamin D ₃ preparations and calcium preparations have been used. In addition, hormone agents such as estrogen preparations and calcitonin preparations have been used for the purpose of suppressing demineralization from bone.

[0007]

However, although the effects of these drugs are found to have some effects in reducing pain, preventing the progress of medical conditions, etc., the effects are not satisfactory, and are more reliable. There is a need for a medicinal component that exhibits various effects. Furthermore, when ingesting the ingredient on a daily basis for the purpose of preventing absorbable bone disease, it is desirable to take it orally, and especially as a blended ingredient of food and drink, it is highly safe that it can be taken with food and drink. Is desired.

[0008]

Means for Solving the Problems The present inventors have previously described PT
We know that calcium and inorganic phosphate release from cultured neonatal mouse calvaria induced by Hrp (parathyroid hormone-related protein) or its active fragments have a high correlation with the mechanism of absorbable bone diseases such as osteoporosis. It was Then, using this method, when searching for various compounds for preventive and therapeutic effects on resorbable bone diseases, hederagenin compounds were found to exhibit a calcium and inorganic phosphate release inhibiting effect in this evaluation system, Filed a patent application (Japanese Patent Application No. 5-17844)
1). Furthermore, the inventors of the present invention, regarding saponin compounds,
When we conducted a thorough screening using this method,
The inventors have found that various glycyrrhetinic acid compounds exhibit a calcium and inorganic phosphate release inhibiting effect in this evaluation system, and completed the present invention.

That is, the present invention is based on the formula (1)

[0010]

[Chemical 2]

(Wherein R represents a hydrogen atom or a glycosyl group) and 3-dehydroglycyrrhetinic acid in which the 3-position thereof is converted to a carbonyl group, or physiologically acceptable thereof. The present invention provides a preventive / therapeutic agent for bone diseases, which contains salt as an active ingredient.

The glycyrrhetinic acid compound used in the present invention means glycyrrhetinic acid [a compound of the formula (1) in which R is a hydrogen atom and the configuration at the 3-position is S], 3-
Epi-glycyrrhetinic acid [a compound in which R is a hydrogen atom in the formula (1) and the configuration at the 3-position is R] and glycyrrhetinic acid glycosides in which the hydrogen atom of the R in the formula (1) is substituted with various glycosyl groups Means the body, for example, the Spanish legume (Glycyrrhiza glab)
Examples thereof include glycosides such as glycyrrhizin which is a saponin component of ra).

Further, as shown in Examples below, since glycyrrhetinic acid itself which is an aglycone exhibits a bone resorption inhibiting action, a partial hydrolyzate of glycyrrhizinate and various glycosyltransferase reactions, etc., cause various glycosyl groups in R. Various glycyrrhetinic acid glycosides having introduced thereinto can also be effectively used in the present invention.

The glycyrrhetinic acid compound used in the present invention is a known compound known as an effective component of "Japanese licorice" in the Japanese Pharmacopoeia, and is used as an expectorant, an antidote, a liver disease agent, an antiallergic agent, an anti-inflammatory agent and the like. Is known,
It has not been found that these compounds have an effect of suppressing bone resorption and are effective for absorbable bone diseases such as osteoporosis.

The glycyrrhetinic acid compound used in the present invention may be, for example, a natural product purified from a plant such as the above-mentioned Spanish leguminous legume or a synthetic product synthesized by a known method. Further, it may be a semi-synthetic product in which various glycosyl groups are introduced into R by various glycosyltransferase reactions or the like to aglycone glycyrrhetinic acid. In addition, when using a natural product, it is not always necessary to purify it to a pure product, and an extract, a crudely purified product, a mixture of various glycyrrhetinic acid compounds, or the like can be used as long as the effects of the present invention are not impaired. . For example, since it is described in the 12th revised Japanese Pharmacopoeia as "liquorice", "liquorice powder", "liquorice extract" and "crude liquorice extract", commercially available products can be used as they are.

The active ingredient of the present invention is a component of a medicinal plant which has been widely used since ancient times, and no toxicity problem is observed in the usual oral administration. For example, the LD _{50 of} intraperitoneal administration of glycyrrhizin, which is one of the active ingredients of the present invention, to mice is 805 mg / kg (Japan Pharmaceutical Collection 3
P. 24, 1991 (Pharmaceutical Journal), LD in the intraperitoneal administration of aglycone glycyrrhetinic acid to mice.
₅₀ is described as about 308 mg / kg (page 325). In addition, it is generally known that saponin compounds exhibit hemolytic properties, and this hemolytic property is lost by removal of glycosyl groups (Bioactive Natural Substances, p. 419, Ito Dental Publishing, 1978), but the active ingredient of the present invention. Glycyrrhizin, which is a type of glycerin, has a weak hemolytic property (see Segal et al., Biochemical Pharmacology 15: 1411 1
966), the glycoside can be used as it is even in the case of transvascular administration.

The preventive / therapeutic agent for absorbable bone diseases of the present invention is
It is manufactured by using the above-mentioned glycyrrhetinic acid compound as an active ingredient and formulating it in combination with a known pharmaceutical carrier according to a conventional method. Further, in the case of producing a preparation by mixing these compounds, if desired, it may be mixed in the form of a physiologically acceptable salt such as sodium salt, potassium salt or ammonium salt of these compounds. It is possible.

The active ingredient of the present invention can be administered in various dosage forms according to a conventional method. For example, as an oral administration agent,
Capsules, tablets, granules, fine granules, syrups, dry syrups and the like can be exemplified. As parenteral administration agents, in addition to injections, suppositories such as suppositories and vaginal suppositories, nasal administration such as sprays Examples thereof include percutaneous absorbents such as agents, ointments and transdermal tapes.

It is also possible to add the active ingredient of the present invention to ordinary foods and drinks and take it on a daily basis. There is no particular limitation on the type of food that can be added, because the active ingredient of the present invention has a unique flavor, for example, beverages such as drinks, or foods with strong sweetness such as candy, the original taste of the food. It is preferable to add about 0.01 to 1% by weight, which is a range not impaired.

When the active ingredient of the present invention is orally administered, the total amount of the active ingredients is usually 1 to 1 per adult per day.
00 mg / kg, preferably 3 to 30 mg / kg
It may be administered in 3 divided doses. In addition, the intake amount when the active ingredient of the present invention is added to foods and drinks and ingested on a daily basis is similar to the dose amount for oral administration. These can be appropriately increased or decreased depending on age, symptoms, and the like.

When the active ingredient of the present invention is administered parenterally, the dose may be about one-tenth that of oral administration in consideration of the blood concentration of the active ingredient. , The total of active ingredients is usually 0.1 to 10 per adult per day.
1 mg / kg, preferably 0.3-3.0 mg / kg
It may be administered in 3 divided doses. These doses can be appropriately increased or decreased depending on age, symptoms and the like.

Next, with respect to the glycyrrhetinic acid compound which is the active ingredient of the present invention, a method for evaluating the calcium and inorganic phosphate release inhibiting action will be described.

PTHrp is a protein identified as a causative agent of human hypercalcemia, and PTHr is a fragment consisting of amino acid residues 1-34.
p (1-34) is the active form. And this PTHr
p (1-34) has been shown to promote bone resorption in vitro (Journal of Clinical Investigation, Vol. 81, No. 2, 596-600).
Page, 1988; End Clinology, Volume 123, 28
41-2848, 1988), it can be used as an evaluation system for osteoporosis.

Therefore, the in vitro bone resorption inhibitory effect of the active ingredient of the present invention was evaluated using cultured newborn mouse calvaria and PTHrp (1-34). This evaluation system is based on the method of Royce using forearm bones of fetal rats and ⁴⁵ Ca (Journal of Clinical Investigation, 44, No. 1, 103-116, 1).
965), and has the advantage that many specimens can be safely evaluated without using radioisotopes.

This improved method has already been widely used as a screening system for therapeutic agents for bone diseases (Journal of the Japanese Society for Bone Metabolism, Vol. 8, No. 3, 221 page, 1990; 9).
Volume 3, Issue 239, 1991; Japanese Journal of Pharmacology, Volume 55, Addendum 1,
120 pages, 1991), but the outline is as follows.

That is, the calvaria of 4 to 6-day-old ICR mice was collected and pre-cultured for 1 day, and then the medium to be tested and the medium to which 10 ⁻⁸ M PTHrp (1-34) had been added were exchanged. After further culturing for 2 days, the calcium and inorganic phosphorus concentrations in the culture supernatant are measured after completion of the culture, and PTHr
This is a method for comparison with the addition group containing only p (1-34).

When the bone resorption inhibitory effect was evaluated by this method, as shown in the Examples below, the group to which the compound of the present invention was added was compared with the group treated with PTHrp (1-34) alone, and the culture supernatant was It was found that the calcium and inorganic phosphorus concentrations in the product were reduced.

[0028]

The glycyrrhetinic acid compound, which is the active ingredient of the present invention, is effective in treating calcium and inorganic matter from the calvaria of newborn mouse cultivated by the parathyroid hormone-related peptide PTHrp (1-34), as shown in the Examples below. It has the effect of suppressing phosphate release. As described above, the glycyrrhetinic acid compound used as an active ingredient in the present invention has an action of suppressing the release of calcium and inorganic phosphate, so that malignant hypercalcemia, Paget's disease of bone, osteoporosis, and other absorbable bone diseases are suppressed. It is presumed that effective prevention / treatment is possible.

[0029]

EXAMPLES Next, the present invention will be described in more detail based on examples, but the present invention is not limited to these examples.

Example 1. By PTHrp (1-34)
Calcium and calcium from evoked cultured neonatal mouse calvaria
Measurement of Inhibitory Effect on Inorganic Phosphorus Release ICR mice (4 to 6 days old) are cut out from the calvaria to remove soft tissue, and then punched out to a diameter of 4 mm. 5% FBS-BGJb medium [BGJb medium containing 5% fetal bovine serum (Fitton-Jackson m
(Origination / Sigma))]
Put each piece in an 8-well plate and pre-culture for 24 hours under the conditions of 37 ° C., 5% carbon dioxide / air. After the pre-culture,
The medium is exchanged with 5% FBS-BGJb medium containing 10 ⁻⁸ M PTHrp (1-34) and the sample having the concentration shown in the table, and the cells are further cultured for 48 hours.

After culturing, VX10 manufactured by JEOL Ltd.
Using an 00 type biochemical automatic analyzer, the calcium concentration of the culture supernatant was measured by the OCPC method, and the inorganic phosphorus concentration was measured by the molybdic acid direct method, and compared with the value of the control group to which only PTHrp (1-34) was added. Then, the bone resorption effect was examined. The results are shown in [Table 1].

[0032]

[Table 1]

As is clear from the results shown in [Table 1], all the glycyrrhetinic acid compounds of the present invention suppressed the release of calcium and inorganic phosphate. From these results, it is strongly suggested that the glycyrrhetinic acid compound of the present invention can prevent or suppress malignant hypercalcemia, and it is clear that the compound of the present invention is useful as a prophylactic / therapeutic agent for bone diseases. Became.

Example 2. Production of capsules [Prescription] Glycyrrhizin 100 parts (parts by weight) Potato starch 148 parts 〃 Magnesium stearate 2 parts 〃

[Manufacturing Method] The above ingredients were well mixed by a muller according to the formulation, and 250 g of No. 1 hard gelatin capsule was mixed.
Each capsule was filled with mg to obtain a capsule containing 100 mg of glycyrrhizin in one capsule.

Example 3. Manufacture of rectal suppository Witepsol H-15 was heated and melted, glycyrrhetinic acid was added to this at a concentration of 12.5 mg / ml and mixed uniformly, and then 2 ml of this was placed in a rectal suppository mold.
Each was injected and cooled to prepare a rectal suppository containing 25 mg of glycyrrhetinic acid in one preparation.

Example 4. Preparation of drink [prescription] Glycyrrhizin 4g DL-sodium tartrate 10mg Succinic acid 1mg Liquid sugar 80g Citric acid 1.2g Vitamin C 1g Flavor 1ml Potassium chloride 0.1g Magnesium sulfate 50mg

[Production Method] The above ingredients were dissolved in 800 ml of distilled water according to the recipe, and distilled water was added to obtain a total volume of 1000 ml.
Then, sterilize with 0.22 μm sterilization filter and
Aseptically fill each 00 ml in a brown bottle, 40 per drug
A drink preparation containing 0 mg of glycyrrhizin was obtained.

Example 5. Production of candy [Prescription] Japanese Pharmacopoeia licorice powder 2 g Purified water 1 g Granulated sugar 49 g Syrup 48 g Citric acid 0.5 g Lemon flavor 0.5 g

[Production Method] Granulated sugar and starch syrup were melted by heating in accordance with a conventional method, and the other components were suspended in purified water and added, and then uniformly mixed to produce a candy of 2 g per grain. Each grain contains 40 mg of licorice powder.

[0041]

The glycyrrhetinic acid compound, which is the active ingredient of the present invention, suppresses the release of calcium and inorganic phosphorus from the calvaria of the cultured neonatal mouse caused by PTHrp (1-34), and thus has a bone resorption effect. It was confirmed to suppress. These active ingredients are ingredients of medicinal plants that have been widely used since ancient times, and there is no particular problem in safety.

Therefore, the preventive / therapeutic agent for absorbable bone diseases of the present invention is useful for the prevention or treatment of absorbable bone diseases such as malignant hypercalcemia, Paget's disease-induced osteoporosis and the like. . Further, by adding the active ingredient of the present invention to food and drink and ingesting it daily, it is also useful as a health food or the like for the prevention of absorbable bone diseases. (that's all)

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[Procedure amendment]

[Submission date] March 17, 1995

[Procedure Amendment 1]

[Document name to be amended] Statement

[Name of item to be corrected] 0032

[Correction method] Change

[Correction content]

[0032]

[Table 1]

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication location A61K 31/70 C07C 62/32 9450-4H C07H 15/256 Z C07J 63/00 9051-4C (72 ) Koichi Nakahara 1-1-1, Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka Prefecture Suntory Biomedical Research Institute

Claims (2)

[Claims] 1. The following formula (1): (In the formula, R represents a hydrogen atom or a glycosyl group.) A prophylactic / therapeutic agent for a bone disease containing a glycyrrhetinic acid compound or 3-dehydroglycyrrhetinic acid or a physiologically acceptable salt thereof as an active ingredient . 2. The preventive / therapeutic agent according to claim 1, wherein the bone disease is malignant hypercalcemia, Paget's disease of bone or osteoporosis.