JPH06329697A - Medicine for improvement of bone disease - Google Patents

Abstract

PURPOSE:To provide a medicine for improvement of bone diseases, containing a bisphosphonated steroid hormone, etc., as the active component, free from decomposition in blood, exhibiting its pharmaceutical activity by selective adsorption to the bone, low in side effect and excellent in water solubility. CONSTITUTION:A bisphosphonated steroid hormone or a bisphosphonated estrogen respectively represented by the formula (AO is a residue of a steroid hormone, a synthetic estrogen, a plant estrogen, etc., respectively having a human bone resorption inhibitory activity or an osteogenesis promotion activity; R<1> is a lower alkylene or a lower hydroxyalkylene; R<2> is H or OH) and/or a salt thereof is used as the active component. An additive composed of lactose, wheat starch, polyethylene glycol, talc, magnesium stearate, etc., is blended therewith and the resultant mixture is dried at 40 deg.C under vacuum. Preparation is subsequently carried out according to the compressing method, etc., thus producing the objective medicine for improvement of bone diseases, capable of being selectively adsorbed without being decomposed in blood and exhibiting a pharmaceutical activity on the bone.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a therapeutic agent for bone diseases,
More specifically, the present invention relates to a therapeutic agent for bone diseases containing a compound in which a steroid hormone or the like is bound to a residue of a bisphosphonated compound and / or a pharmaceutically acceptable salt thereof.

[0002]

2. Description of the Related Art Conventionally, for the treatment of bone diseases, in addition to administration of calcium agents, active vitamin D, intramuscular injection of calcitonin preparations, steroidal hormones such as 17β-estradiol and norethandrolone, synthetic estrogens, and plant estrogens. Etc. are used. However, in this case, side effects such as the expression of female hormone action on the uterus and liver damage have been reported.

Therefore, in order to avoid the above-mentioned side effects, tetracycline, polymalonate or phosphonates, which are well known as compounds having affinity for bone, are combined with the above-mentioned steroid hormones, and the steroid hormones are added to the uterus. Attempts have been made to selectively accumulate in the bone rather than.

However, also in this case, depending on the binding method, the binding substance of the osteophilic compound and the steroid hormone is broken by the action of the degrading enzyme in the blood, and the released steroid hormone is released to the bone. It may not be able to adsorb selectively. If this is not the case, the two must be combined in a special way.

An example of such an attempt is disclosed in Japanese Patent Laid-Open No. 4-3.
52795, amino or hydroxylalkyl-
It has been reported as a novel bone agonist that has a local therapeutic effect on bone in which 1,1-bisphosphonic acid and a hydroxy-containing steroidal hormone are bound. However, it is actually adsorbed to bone selectively. It is not clear whether it exhibits its pharmacological action. Further, there remains a problem in terms of solubility in water.

[0006]

SUMMARY OF THE INVENTION The present invention has been made from the above viewpoint, and is not decomposed in blood,
It is an object of the present invention to provide a safe therapeutic agent for bone diseases that selectively adsorbs to bone, can sufficiently exert a pharmacological effect on bone, and has few side effects, and further, a therapeutic agent for bone diseases having excellent solubility in water. To do.

[0007]

Means for Solving the Problems As a result of intensive research to solve the above problems, the present inventor has found that bisphosphonated compounds and steroid hormones, synthetic estrogens, phytoestrogens and the like have alkylene groups or If linked by an ether bond using a hydroxyalkylene group, this compound will be selectively carried to the bone without being decomposed in blood and adsorbed to the bone, and part of the compound will be broken to release the steroid hormone. It is found that not only the bone resorption inhibiting action but also the bone formation promoting action is exerted on the bone, and the bisphosphonated compound is a hydroxyalkyl-1,1, -diphosphonic acid. In that case, in addition to the above characteristics, it was found that it is also excellent in water solubility
The present invention has been completed.

That is, the present invention is a therapeutic agent for bone diseases, which comprises a compound represented by the structural formula of Chemical formula 2 and / or a pharmaceutically acceptable salt thereof as an active ingredient.

[0009]

[Chemical 2]

However, in the chemical formula 2, A-O- is one residue selected from steroid hormones having human bone resorption inhibitory activity or osteogenesis promoting activity, synthetic estrogen, and plant estrogen, and R ¹ is , A lower alkylene group or a lower hydroxyalkylene group, and R ² represents a hydrogen atom or a hydroxyl group.

The present invention will be described in detail below. The therapeutic agent for bone diseases of the present invention contains, as an active ingredient, one or more of the compound represented by the chemical formula 2 and / or a pharmaceutically acceptable salt thereof.

In the above chemical formula 2, R ¹ represents a lower alkylene group or a lower hydroxyalkylene group, and the carbon number of these groups is preferably 1 to 10, more preferably 1 to 6, More preferably, it is 1-4. Also,
The number of hydroxyl groups contained in the lower hydroxyalkylene group is not particularly limited and is 1 or 2 or more.

Of the compounds represented by the above chemical formula 2, R
^The compound in which ¹ is a lower hydroxyalkylene group or the compound in which R ² is a hydroxyl group has particularly good solubility in water and is preferably used in the present invention together with a pharmaceutically acceptable salt of these compounds.

Examples of the steroid hormones include anabolic steroids, male hormones, estrogen hormones, luteinizing hormones and the like, and specifically 17β-estradiol, testosterone, norethandrolone, nandrolone, tibolone and the like.

The synthetic estrogens include stilbestrol, monomestrol, hexestrol, promethestrol, dienestrol, benzestrol. )
However, as the plant estrogens, genistein (Ge
nistein), biochanin (Biochanin), formononetin (Formononetin), daidzoin (Daidzoin), cameltrol (Canmestrol), miroestrol (Miroestr)
ol) is preferred.

In the following, as an example of the compound represented by the above chemical formula 2, the moiety represented by A--O-- is a residue obtained by reacting either one of the two hydroxyl groups of 17β-estradiol. The structure of the compound is shown below.

[0017]

[Chemical 3]

[0018]

[Chemical 4]

[0019] However, in Formula 3 and of 4, R ¹ is a lower alkylene group or a lower hydroxyalkylene group (one or more has been substituted with a hydroxyl group based on the hydrogen atoms of the lower alkylene group), R ² Represents a hydrogen atom or a hydroxyl group, respectively.

The method for producing these compounds will be described with reference to the reaction formula shown in Chemical formula 5, taking the case where R ^{2 of the} compound shown in Chemical formula 3 is a hydrogen atom as an example.

[0021]

[Chemical 5]

In the chemical formula 5, R ¹ represents a lower alkylene group or a lower hydroxyalkylene group, X represents a halogen atom, and Bn represents a benzyl group. In addition, in compound 3 and compound 4, when R ¹ is a lower hydroxyalkylene group, the hydroxyl group thereof is t-BuMe.
₂ Protected by Si (tert-butyldimethylsilyl) group.

DMF (Dimethylformamide) of NaH
A solution of 17β-estradiol (Compound 1) in DMF was added dropwise to the solution under ice-cooling, an alkyl halide was added, and the mixture was stirred at room temperature for 4 hours, and the obtained reaction product was extracted with ethyl acetate or the like, followed by ethyl acetate. Crystallize with hexane or the like to obtain compound 2. CH ₂ C of this compound 2 and 2,4-lutidine
t-BuMe ₂ SiOTf (tert-butyldimethylsilyltrifluoromethanesulfonate) was added dropwise to the ₁₂ solution under ice cooling, and the mixture was stirred at room temperature for 1 hour, and the obtained reaction product was extracted with ethyl acetate or the like. After that, it is purified by silica gel column chromatography or the like to obtain compound 3.

Next, the DMF solution of the phosphoric acid compound was added dropwise to the DMF solution of KH under ice cooling, and after stirring for 30 minutes, the DMF solution of the compound 3 was added dropwise and stirred at 30 ° C. for 9 hours to obtain The obtained reaction product is extracted with ethyl acetate or the like and then purified by silica gel column chromatography or the like to obtain compound 4. This compound 4 was added to a mixed solution of 1N-hydrochloric acid and methanol and stirred at room temperature for 4 hours. The obtained reaction product was extracted with ethyl acetate or the like, and then purified by preparative silica gel thin layer chromatography or the like to give compound 5 To get Finally, the compound 5 was dissolved in methanol, Pd (OH) ₂ was added, and the mixture was stirred under a hydrogen stream at room temperature for 1 hour. The obtained reaction product was purified to obtain a compound 6 (in the formula 3, R ² is a hydrogen atom). Compound) is obtained.

Further, the case where R ² is a hydroxyl group in the compound represented by Chemical formula 3 will be described using the reaction formula shown in Chemical formula 6.

[0026]

[Chemical 6]

However, in the chemical formula 6, R ¹ represents a lower alkylene group or a lower hydroxyalkylene group, R ³ represents an alkyl group, and X represents a halogen atom. Compound 8
In Compound 12, when R ¹ is a lower hydroxyalkylene group, the hydroxyl group is t-BuMe.
₂ Protected by Si group.

To a solution of NaH in DMF under ice-cooling, 17β
-Dripping a solution of estradiol (Compound 1) in DMF
Afterwards, the alkyl halide with carboxylic acid ester
The mixture was added and stirred at room temperature for 4 hours, and the obtained reaction product was extracted with ethyl acetate.
Etc., then crystallize with ethyl acetate-hexane, etc.
Obtain compound 7. CH of this compound 7 and 2,4-lutidine
₂Cl₂The solution was added to t-BuMe under ice cooling.₂SiOTf
After the dropping, the mixture was stirred at room temperature for 1 hour, and the obtained reaction product was mixed with acetic acid.
After extracting with chill, etc., silica gel column chromatograph
And the like to obtain compound 8.

This compound 8 was reacted with sodium hydroxide to give compound 9, which was added to a benzene solution of compound 9 at room temperature.
MF and xazalyl chloride were added dropwise, the mixture was stirred for 30 minutes, and then the solvent was evaporated under reduced pressure to give compound 10 (acid chloride form).
And This compound 10 is dissolved in anhydrous ether under a stream of argon, and triisopropylphosphite is added dropwise under ice cooling. After stirring under ice-cooling for 1 hour, room temperature for 18 hours, and further at 45 ° C. for 30 minutes, the solvent was distilled off under reduced pressure to obtain an acylphosphonoester compound 11.

Under an argon stream, diisopropyl phosphite and a diethylamine anhydrous ether solution were cooled with ice,
Anhydrous ether solution of compound 11 is added dropwise and stirred for 2 hours and a half. The solvent is distilled off under reduced pressure, and the residue is purified by silica gel chromatography or the like to obtain compound 12. This compound 12
Is added to a mixed solution of 1N-hydrochloric acid and methanol, the mixture is stirred at room temperature for 4 hours, the obtained reaction product is extracted with ethyl acetate and the like, and then purified by preparative silica gel thin layer chromatography and the like to obtain Compound 13.

Trimethylsilyl bromide was added dropwise to an anhydrous chloroform solution of compound 13 under an argon stream under ice cooling, and the mixture was stirred under ice cooling for 1 hour and at room temperature for 20 hours. After adding water under ice cooling and stirring at room temperature for 1 hour, the reaction solution was concentrated under reduced pressure, and the obtained powder was washed with benzene or the like to give Compound 1
4 (a compound in which R ² is a hydroxyl group in Chemical formula 3) is obtained.

The residue of the bisphosphonated compound can be introduced through the phenolic hydroxyl groups of synthetic estrogens and phytoestrogens in the same manner as above.

Next, of the compound represented by Chemical formula 4, which is another example, a method for producing a compound in which R ² is a hydrogen atom will be described using the reaction formula shown in Chemical formula 7.

[0034]

[Chemical 7]

In the chemical formula 7, R ¹ represents a lower alkylene group or a lower hydroxyalkylene group having a hydroxyl group protected by a benzyl group, etc., and Bn represents a benzyl group. In the compound 22, R ¹ represents a lower alkylene group or a lower hydroxyalkylene group.

Estrone (Compound 15) was added to a DMF solution of NaH under ice-cooling, stirred for 10 minutes, benzyl chloride was added dropwise, and the mixture was stirred at room temperature for 1 hour, and the obtained reaction product was extracted with ethyl acetate. After that, the compound 16 is obtained by crystallization with chloroform-hexane or the like. Compound 16, dihydric alcohol, p-toluenesulfonic acid hydrate and benzene are mixed, heated and stirred, and reacted for 16 hours while separating water using a Dean-Stark tube. The obtained reaction product is extracted with ethyl acetate and then crystallized with chloroform-hexane to give compound 17.

AlCl ₃ is added to THF (tetrahydrofuran) at 0 ° C. under an argon stream, LiAlH ₄ is then added little by little, and a THF solution of compound 17 is further added dropwise, followed by stirring under reflux for 3 hours. This reaction solution was diluted with ether, Na ₂ SO ₄ · 10H ₂ O was added, and the mixture was stirred for 12 hours.
The precipitated Al (OH) ₃ was filtered off and the compound 18 was collected from the filtrate.
To get Methanesulfonyl chloride was added dropwise to a pyridine solution of this compound 18 under ice cooling, and the mixture was stirred at room temperature for 2 hours,
After extraction with ethyl acetate, crystallization with ethyl acetate-hexane or the like gives compound 19. This compound 19 and NaI
Is dissolved in diglyme, stirred under reflux for 1 hour, extracted with ethyl acetate, and then crystallized with ethyl acetate-hexane or the like to obtain compound 20.

Next, the DMF solution of the phosphoric acid compound was added dropwise to the DMF solution of KH under ice cooling and stirred for 30 minutes, and then the DMF solution of the compound 20 was added dropwise and stirred for 48 hours at 30 ° C., and acetic acid was added. After extraction with ethyl or the like, purification by silica gel column chromatography or the like gives compound 21. To a methanol solution of this compound 21, Pd (OH) ₂ was added,
The mixture is stirred under a hydrogen stream at room temperature for 4 hours, and the obtained reaction product is purified to obtain a compound 22 (a compound in which R ² is a hydrogen atom in Chemical formula 4).

Further, among the compounds represented by Chemical formula 4, R ²
A method for producing a compound in which is a hydroxyl group will be described using the reaction formula shown in Chemical formula 8.

[0040]

[Chemical 8]

In the chemical formula 8, R ⁴ represents a lower alkylene group or a lower hydroxyalkylene group having a hydroxyl group protected by a benzyl group, etc., and Bn represents a benzyl group. In addition, in the compound 28 and the compound 29,
R ⁴ represents a lower alkylene group or a lower hydroxyalkylene group. In addition, the compound 23 as a starting material is a compound in which R ¹ is R ⁴ .CH ₂ among the compounds 18 in the above chemical formula 7, and this compound is obtained by the same production method as the above production method. is there.

1. Acetone solution of compound 23 under ice cooling, 2.
Add 8N Jones reagent (chromium (VI) oxide-sulfuric acid) until it becomes orange-green, and stir for 1.5 hours. Isopropyl alcohol is added to the reaction solution to decompose excess oxidizing agent, water is added to the reaction solution, extraction is performed with ethyl acetate, washing with water and dehydration (Na ₂ SO ₄ ), and then the solvent is distilled off. The residue is purified by silica gel column chromatography or the like to obtain compound 24.

DM in benzene solution of compound 24 at room temperature
F and xazalyl chloride are added dropwise, the mixture is stirred for 30 minutes, and then the solvent is distilled off under reduced pressure to give compound 25 (acid chloride form). This compound 25 was dissolved in anhydrous ether and anhydrous methylene chloride under an argon stream, and triisopropyl phosphite was added dropwise under ice cooling, under ice cooling for 1 hour, at room temperature for 18 hours, and further at 45 ° C. for 30 minutes. After stirring, the solvent is distilled off under reduced pressure to give compound 26 (acylphosphonoester form).

In an anhydrous ether solution of diisopropyl phosphite and diethylamine under an argon stream under ice cooling,
Anhydrous ether solution of compound 26 was added dropwise and stirred for 2 hours and a half, then the solvent was distilled off under reduced pressure and the residue was purified by silica gel chromatography to obtain compound 27.

Pd (O
H) ₂ was added, and the mixture was stirred at room temperature for 20 minutes under a hydrogen stream, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give compound 2
Get 8. Trimethylsilyl bromide was added dropwise to an anhydrous chloroform solution of compound 28 under an argon stream under ice cooling, and the mixture was stirred under ice cooling for 1 hour and at room temperature for 20 hours. Water was added thereto under ice cooling, and the mixture was stirred at room temperature for 1 hour, then the reaction solution was concentrated under reduced pressure, and the obtained powder was washed with benzene or the like,
A compound 29 (a compound in which R ² is a hydroxyl group in Chemical formula 4) is obtained.

Further, the above-mentioned production method exemplifies the production process of the compound represented by the above-mentioned chemical formula 2 used in the present invention, which limits the production method such as the starting material or the processing conditions. Not something.

The therapeutic agent for bone diseases of the present invention contains one or more of the above compounds and / or pharmaceutically acceptable salts thereof. The dosage form of the therapeutic agent for bone diseases of the present invention is
Although not particularly limited, generally harmless one or several types of vehicles, carriers, excipients, integrators, preservatives, stabilizers, flavoring agents and the like, which are generally acceptable in terms of formulation, are mixed, and tablets and granules are prepared. ,
It may be an internal preparation such as a capsule or a drench, an external preparation such as an ointment, a cream or a liquid, an injectable preparation such as a sterile solution or suspension. These can be manufactured using a conventionally known technique.

For example, the above compounds and / or salts thereof and a binder such as corn starch and gelatin, an excipient such as microcrystalline cellulose, a potato starch, a puffing agent such as sodium alginate and a sweetener such as lactose and sucrose. Distribute the powder,
It can be a tablet, granule, or capsule. Also,
In the case of an injection, distilled water for injection, polyethylene glycol or the like is used as a solvent, or a dispersant, a buffer, a preservative, an antioxidant or the like may be added thereto as required. When used as an external preparation, petrolatum, paraffin, oils and fats, lanolin and the like can be used as a base.

Among the above compounds, when a compound in which R ¹ is a hydroxylalkylene group and / or a salt thereof or a compound in which R ² is a hydroxyl group and / or a salt thereof is used, these are excellent in water solubility, and therefore, a preparation In many cases, it is advantageous in terms of conversion.

The dose varies depending on the age of the patient, the type of disease, the symptoms, etc., but generally, for oral administration per adult per day, the amount of the above compound or its salt is 0.1 mg to 10 g 0.01 mg for agents
~ 1 g, and by using as an external preparation in the range of 0.1 mg to 50 g, expected effects can be expected.

[0051]

EXAMPLES Examples of the present invention will be described below. First, production examples of the compound used in the present invention will be explained.

[0052]

[Production Example 1] 2.4 g of 55% NaH in 20 mL of DM
To the solution dissolved in F, add 1 mL to 30 mL DMF under ice cooling.
A solution in which 10 g of 7β-estradiol (the compound represented by 1 in Chemical formula 5) was dissolved was added dropwise. After the dropping is completed,
37.4 mL of 1,3-dibromopropane was added, and the mixture was stirred at room temperature for 4 hours. Then, the reaction solution was poured into ice water and extracted with ethyl acetate, the obtained extract was washed with water, dehydrated with MgSO _4, and then the solvent was distilled off. Furthermore, under reduced pressure,
Excess 1,3-dibromopropane was distilled off, and the residue was crystallized from ethyl acetate-hexane to give 3- (3-bromopropyloxy) -17β-hydroxy-estra-1,
3,5 (10) -triene (of the compounds shown by 2 in the above chemical formula 5, X = Br, R ¹ = (CH ₂ ) ₃ ) 1
1.4 g (yield 78.8%) was obtained.

The above 3- (3-bromopropyloxy)-
17β-hydroxy-oestra-1,3,5 (10)-
200 mg triene and 0.12 mL 2,4-lutidine
1 mL of CH₂Cl₂Under ice-cooling,
0.175 mL t-BuMe ₂After dropping SiOTf,
Stirred at room temperature for 1 hour. Then, pour the reaction solution into ice water,
Extract with ethyl acetate and wash the resulting extract with MgS
O_FourAfter dehydration with, the solvent is distilled off and the residue is filtered.
Purified by silica gel column chromatography, 3- (3
-Bromopropyloxy) -17β-tert-butyl
Rudimethylsilyloxy-estradio-1,3,5 (1
0) -triene (of the compounds represented by 3 in Chemical formula 5 above)
X = Br, R¹= (CH₂)₃Compound) 190 mg (yield
74.5%).

340 mg of 35% KH in 5 mL of DMF
A solution of 1.25 g of tetrabenzyl diphosphonate in 5 mL DMF was added dropwise to the solution dissolved in 5 mL of DMF, and the mixture was stirred for 30 minutes, and then the above-mentioned 3- (3-bromopropyloxy) -17β-. A solution of 1.0 g of tert-butyldimethylsilyloxy-estra-1,3,5 (10) -triene dissolved in 5 mL of DMF was added dropwise.
The mixture was stirred at 30 ° C for 9 hours. After that, the reaction solution is poured into water,
Extract with ethyl acetate and wash the resulting extract with MgS
After dehydration with O ₄ , the solvent was distilled off and the residue was purified by silica gel column chromatography to give 3-
(4,4-Diphosphonobutyloxy) -17β-tert-butyldimethylsilyloxy-estradiol
1,3,5 (10) -triene tetrabenzyl ester (of the compounds represented by 4 in the above Chemical formula 5, R ¹ = (C
H ₂ ) ₃ compound) (980 mg, yield 51.6%) was obtained.

200 mg of the above 3- (4,4-diphosphonobutyloxy) -17β-tert-butyldimethylsilyloxy-estra-1,3,5 (10) -triene tetrabenzyl ester was added to 1 N-hydrochloric acid 0.3 m.
The mixture was added to a mixed solution of L and methanol (4 mL) and stirred at room temperature for 4 hours. Then, the reaction solution was poured into ice water, extracted with ethyl acetate, the obtained extract was washed with water, dehydrated with MgSO ₄ , the solvent was distilled off, and the residue was subjected to preparative silica gel thin layer chromatography. And purified with 3- (4,4-diphosphonobutyloxy) -17β-hydroxy-estra-1,3,5 (10) -triene tetrabenzyl ester (of the compounds shown by 5 in the above Chemical Formula 5, R ¹ =
(CH ₂ ) ₃ compound) (153 mg, yield 86.5%) was obtained.

The above 3- (4,4-diphosphonobutyloxy) -17β-hydroxy-estra-1,3,5
(10) -triene tetrabenzyl ester 830 m
g in 30 mL of methanol, 150 mg
Of Pd (OH) ₂ was added, and the mixture was stirred under a hydrogen stream at room temperature for 1 hour. After that, DMF was added to the reaction solution, and Pd (O
H) ₂ was filtered off. The residue obtained by concentrating the filtrate was washed with methanol to give 3- (4,4-diphosphonobutyloxy) -17β-hydroxy-estra-1,3,5.
380 mg of (10) -triene (a compound of R ¹ = (CH ₂ ) ₃ among the compounds represented by 6 in Chemical formula 5) (yield 7
9.4%). The melting point of the obtained sample is 239 to 24
It was 0 ° C. In addition, the nuclear magnetic resonance (NM
R) shows the spectrum.

NMR δ (DMSO-d ₆ ) 0.67 (3H, s, 18-H ₃ ) 2.64 to 2.80 (2H, m, 6-H ₂ ) 3.49 (1H, t, J = 5 Hz, 17α-H) 3.80 to 3.91 (2H, m, -OCH _2- ) 6.59 (1H, d, J = 2Hz, 4-H) 6.65 (1H, dd, J = 9) , 2Hz, 2-H) 7.13 (1H, d, J = 9Hz, 1-H)

[0058]

[Production Example 2] A solution prepared by dissolving 1.1 g of NaH in 15 mL of DMF was added with estrone (15 in Chemical Formula 7 above) under ice cooling.
Was added thereto, and the mixture was stirred for 10 minutes, then 2.8 mL of benzyl chloride was further added dropwise, and the mixture was stirred at room temperature for 1 hour. Then, the reaction solution was poured into ice water, extracted with ethyl acetate, the obtained extract was washed with water, dehydrated with MgSO ₄ , the solvent was distilled off, and the residue was crystallized with chloroform-hexane to give 3- Benzyloxy-estra-1,
3,5 (10) -trien-17-one (1 in Chemical Formula 7 above)
8.7 g (yield 98.0%) of the compound represented by 6) was obtained.

8.0 g of the above 3-benzyloxy-oestra-1,3,5 (10) -trien-17-one, 2
7.2 mL of ethylene glycol, 420 mg of p-toluenesulfonic acid hydrate and 200 mL of benzene were mixed, heated and stirred, and reacted for 16 hours while separating water using a Dean-Stark tube. After cooling, the reaction solution was poured into ice water, extracted with ethyl acetate, the obtained extract was washed with water,
After dehydration with MgSO ₄ , the solvent was distilled off and the residue was crystallized from chloroform-hexane to give 3-benzyloxy-17,17-ethylenedioxy-estradiol.
8.1 g of 1,3,5 (10) -triene (a compound of R ¹ = (CH ₂ ) ₂ among the compounds represented by 17 in the above chemical formula 7)
(Yield 90.2%) was obtained.

400 mL of THF (tetrahydrofuran
40 g of AlCl 2 at 0 ° C. under an argon stream.₃Add
Then 6.8g LiAlH_FourWas added little by little.
Furthermore, 8.0 g of the above 3-benzyloxy was added to the reaction solution.
-17,17-ethylenedioxy-estra-1,3
The 5 (10) -triene was dissolved in 200 mL of THF.
The solution was added dropwise and stirred under reflux for 3 hours. After standing to cool, react
Dilute the solution with ether and add Na ₂SO_Four・ 10H₂Add O 1
Precipitated Al (OH) after stirring for 2 hours₃Filtered off and the filtrate
Was concentrated to give a residue, which was crystallized from ethyl acetate-hexane.
3-benzyloxy-17β- (2-hydroxy
Ethyloxy) -oestra-1,3,5 (10) -tri
Ene (of the compounds represented by 18 in the above Chemical Formula 7, R¹=
(CH₂)₂Compound) was obtained (yield 87.0%)
It was

7.0 g of the above 3-benzyloxy-17
β- (2-hydroxyethyloxy) -oestra-1,
To a solution of 3,5 (10) -triene in 15 mL of pyridine, 1.68 mL of methanesulfonyl chloride was added dropwise under ice cooling, and the mixture was stirred at room temperature for 2 hours. Then, the reaction solution was poured into ice water, extracted with ethyl acetate, the obtained extract was washed with water, dehydrated with MgSO ₄ , the solvent was distilled off, and the residue was crystallized with ethyl acetate-hexane, 3-
Benzyloxy-17β- (2-methanesulfonyloxyethyloxy) -oestra-1,3,5 (10) -triene (in the compounds represented by 19 in the chemical formula 7, R ¹ =
(CH ₂ ) ₂ compound) (7.35 g, yield 88.2%) was obtained.

7.3 g of the above 3-benzyloxy-17
β- (2-Methanesulfonyloxyethyloxy) -oestra-1,3,5 (10) -triene and 6.8 g of NaI were dissolved in 300 mL of diglyme, and refluxed for 1
Stir for hours. After that, the solvent was distilled off from the reaction solution, the obtained residue was extracted with ethyl acetate, washed with water, and washed with MgSO 4.
After dehydration with ₄ , the solvent was distilled off and the residue was crystallized from ethyl acetate-hexane to give 3-benzyloxy-1.
7β- (2-iodoethyloxy) -estra-1,
7.30 g of 3,5 (10) -triene (a compound of R ¹ = (CH ₂ ) ₂ among the compounds represented by 20 in Chemical formula 7)
(Yield 93.6%) was obtained.

1.0 g of 35% KH in 15 mL of DMF
3.75 g of tetraben was added to the solution dissolved in
Dilmethylene diphosphonate in 20 mL DMF
The dissolved solution was added dropwise and stirred for 30 minutes. Furthermore, anti
3 g of the above 3-benzyloxy-17β- (2-
Iodoethyloxy) -estra-1,3,5 (10)
-Dripping a solution of triene in 20 mL DMF
And stirred at 30 ° C. for 48 hours. After that, the reaction solution in water
Pour, extract with ethyl acetate, wash the resulting extract with water,
MgSO_FourAfter dehydration using
The residue was purified by silica gel column chromatography and 3
-Benzyloxy-17β- (3,3-diphosphono
Propyloxy) -oestra-1,3,5 (10) -to
Liene tetrabenzyl ester (shown as 21 in Chemical Formula 7 above)
R of the compounds¹= (CH₂) ₂Compound) 2.2
g (yield 40.3%) was obtained.

2.2 g of the above 3-benzyloxy-17
β- (3,3-diphosphonopropyloxy) -estra-1,3,5 (10) -triene Tetrabenzyl ester was dissolved in 50 mL of methanol, and P was added to the solution.
Add 200 mg of d (OH) _2, and add 4 at room temperature under hydrogen flow.
Stir for hours. Then, the reaction solution was filtered, and the residue obtained by concentrating the filtrate was crystallized from ethanol-hexane and recrystallized from acetonitrile-methanol to give 3-hydroxy-17β- (3,3-diphosphonopropyloxy). ) -Estra-1,3,5 (10) -triene (a compound of R ¹ = (CH ₂ ) ₂ among the compounds represented by 22 in the above chemical formula 7) 350 mg (yield 31.5%) was obtained. The melting point of the obtained sample was 244-245 ° C. In addition, in elemental analysis, C ₂₁ H ₃₂ O ₈ P ₂ (3-hydroxy-17β
-(3,3-Diphosphonopropyloxy) -estra-1,3,5 (10) -triene) C: 5
The analytical values of the obtained sample were C: 52.78 and H6.72 with respect to 2.22 and H: 6.81. Further, the NMR spectrum of the sample was measured. The results are shown below.

NMR δ (DMSO-d ₆ ) 0.82 (3H, s, 18-H ₃ ) 2.60 to 2.75 (2H, m, 6-H ₂ ) 3.38 (1H, t, J = 6 Hz, 17α-H) 3.45 to 3.68 (2H, m, -OCH _2- ) 6.49 (1H, d, J = 2Hz, 4-H) 6.54 (1H, dd, J = 9) , 2Hz, 2-H) 7.07 (1H, d, J = 9Hz, 1-H)

[0066]

MANUFACTURING EXAMPLE 3 The same procedure as in Manufacturing Example 2 was repeated except that trimethylene glycol was used in the same molar amount as the above ethylene glycol instead of the ethylene glycol used in the reaction in the above Manufacturing Example 2, and 3-hydroxy was used. -17β-
(4,4-diphosphonobutyloxy) -estra-
1,3,5 (10) -triene (a compound of R ¹ = (CH ₂ ) ₃ among the compounds represented by 22 in the above Chemical formula 7) was obtained.
The melting point of the obtained sample was 241 to 243 ° C. The NMR spectrum of the sample was as follows.

NMR δ (DMSO-d ₆ ) 0.70 (3H, s, 18-H ₃ ) 2.78 to 2.87 (2H, m, 6-H ₂ ) 3.25 (1H, t, J = 6Hz, 17α-H) 3.30 to 3.41 (2H, m, -OCH _2- ) 6.71 (1H, d, J = 2Hz, 4-H) 6.79 (1H, dd, J = 9) , 2Hz, 2-H) 7.20 (1H, d, J = 9Hz, 1-H)

[0068]

Production Example 4 The intermediate substance obtained in Production Example 3 above, 3-benzyloxy-17β- (3-hydroxypropyloxy) -estradi-1,3,5 (10) -triene (indicated by 23 in Chemical Formula 8 above). 2.10 g of the compound of R ⁴ = (CH ₂ ) ₂ among the prepared compounds was dissolved in 15 mL of acetone, and 2.8 N Jones reagent (chromium oxide (V
I) -sulfuric acid) was added until it became orange-green, and the mixture was stirred for 1 hour and a half. Isopropyl alcohol was added to the reaction solution to decompose excess oxidizing agent. Water was further added to this reaction solution, which was extracted with ethyl acetate, washed with water and dehydrated (Na ₂ SO ₄ ), and then the solvent was distilled off.

The residue was purified by silica gel column chromatography (chloroform / methanol = 30/1) and 3
-Benzyloxy-17β- (2-carboxyethyloxy) -estradi-1,3,5 (10) -triene (R ⁴ = (CH ₂ ) ₂ among the compounds represented by 24 in the above chemical formula 8)
1.25 g as a colorless solid (yield 57%)
Obtained.

Here, a part of this was recrystallized from ethyl acetate-n-hexane to purify it as colorless needle-like crystals, and the characteristics were examined using this as a sample. First, the melting point of the obtained sample was 116 to 117 ° C. In addition, in elemental analysis, C ₂₈ H ₃₄ O ₄ (3-benzyloxy-17β- (2
-Carboxyethyloxy) -estra-1,3,5
(10) -Triene) theoretical value C: 77.39, H:
The analytical value of the obtained sample is C: 77.
21, H: 7.93 was obtained. further,
The NMR spectrum of the sample was measured. The results are shown below.

NMR δ (CDCl ₃ ) 0.78 (3H, s), 1.13 to 1.75 (10H,
m) 1.86 (1H, m), 1.98 to 2.32 (4H,
m) 2.63 (2H, t, J = 6.2 Hz), 2.81
2.87 (2H, m) 3.44 (1H, t, J = 8.1Hz), 3.70 ~
3.84 (2H, m) 5.02 (2H, s), 6.70 (1H, d, J = 2.
7 Hz) 6.77 (1H, dd, J = 9.1, 2.0 Hz) 7.19 (1H, d, J = 9.1 Hz), 7.28-
7.45 (5H, m)

The above 3-benzyloxy-17β- (2-
Carboxyethyloxy) -estra-1,3,5 (1
217 mg of 0) -triene was dissolved in 5 mL of benzene, 1 drop of DMF and 0.088 mL of xazalyl chloride were added dropwise at room temperature, and the mixture was stirred for 30 minutes. The solvent was distilled off under reduced pressure and the acid chloride compound (3-benzyloxy-17β-
(2-chlorocarbonylethyloxy) -estradi
Thus, 1,3,5 (10) -triene (a compound of R ⁴ = (CH ₂ ) ₂ among the compounds represented by 25 in the above chemical formula 8) was obtained.

This 3-benzyloxy-17β- (2-
Chlorocarbonylethyloxy) -oestra-1,3
5 (10) -Triene (acid chloride form) was dissolved in 5 mL of anhydrous ether and 2.5 mL of anhydrous methylene chloride under an argon stream, and 0.343 mL of triisopropylphosphite was added dropwise thereto under ice cooling. This solution was cooled under ice for 1 hour, at room temperature for 18 hours, and at 45 ° C for 30 hours.
After stirring for a minute, the solvent was distilled off under reduced pressure, and the acylphosphonoester compound (3-benzyloxy-17β- (2-
Phosphonocarbonylethyloxy) -Estra-
1,3,5 (10) -triene diisopropyl ester (of the compounds represented by 26 in the above chemical formula, R ⁴ = (C
H ₂ ) ₂ compound)) was obtained.

Under a stream of argon, a solution of 0.249 mL diisopropyl phosphite and 0.041 mL diethylamine in 2 mL anhydrous ether was cooled with ice,
The above 3-benzyloxy-17β- (2-phosphonocarbonylethyloxy) -oestra-1,3,5 (1
A solution of 0) -triene diisopropyl ester (acylphosphonoester form) dissolved in 2.4 mL of anhydrous ether was added dropwise, and the mixture was stirred for 2.5 hours. The solvent was distilled off from this reaction solution under reduced pressure, and the residue was purified by silica gel chromatography (chloroform / methanol = 60/1) to give 3-benzyloxy-17β- (3,3-diphosphono-3-hydroxypropyl). Oxy) -estra-1,3,5 (10) -triene tetraisopropyl ester (of the compounds represented by 27 in the above Chemical Formula 8, R
⁴ = (CH ₂ ) ₂ compound) as a colorless oil 233 m
g (yield 62%) was obtained. As a result of mass spectrometric analysis of the obtained sample, the mass spectrum (FAB) was m / z: 749 (M
H ⁺ ). Further, in the infrared (IR) Measurement results of the spectrum characteristic absorption of phosphonate was observed at 1250 cm ^-1 or 990 cm ^-1. Further, the NMR spectrum of the sample was as follows.

NMR δ (CDCl ₃ ) 0.77 (3H, s), 1.11 to 1.74 (34H, 34H,
m) 1.85 (1H, m), 1.98 to 2.42 (6H,
m) 2.81 to 2.87 (2H, m), 3.46 (1H,
t, J = 7.8 Hz) 3.98 (2H, t, J = 5.4 Hz), 4.77-
4.94 (4H, m) 5.02 (2H, s), 6.71 (1H, d, J = 2.
7 Hz) 6.77 (1H, dd, J = 8.3, 2.7 Hz) 7.18 (1H, d, J = 8.3 Hz) 7.28 to 7.
45 (5H, m)

3-benzyloxy-17 obtained above
β- (3,3-diphosphono-3-hydroxypropyloxy) -estra-1,3,5 (10) -triene Tetraisopropyl ester 230 mg was dissolved in 5 mL of methanol, and Pd (OH) ₂ 17 mg was added to the solution. The mixture was stirred for 20 minutes at room temperature under a hydrogen stream. The reaction solution is filtered, and the filtrate is concentrated under reduced pressure to give 3-hydroxy-17β.
-(3,3-Diphosphono-3-hydroxypropyloxy) -oestra-1,3,5 (10) -triene
178 mg (yield 87%) of tetraisopropyl ester (a compound of R ⁴ = (CH ₂ ) ₂ among the compounds represented by 28 in Chemical formula 8 above) was obtained as a colorless oil.

As a result of mass spectrometric analysis of the obtained sample, the mass spectrum (FAB) was m / z: 659 (MH ⁺ ). In addition, the measurement result of the IR spectrum shows 1223c
Characteristic absorption of phosphonate was confirmed at m ^-1 and 996 cm ^-1 . Further, the NMR spectrum of the sample was as follows.

NMR δ (CDCl ₃ ) 0.73 (3H, s), 1.06 to 2.43 (41H, 41H,
m) 2.74-2.82 (2H, m), 3.43 (1H,
t, J = 8.2 Hz) 3.99 (2H, t, J = 5.5 Hz), 4.75-
4.94 (4H, m) 5.04 (1H, t, J = 8.2Hz, -C (OH)
(PO ₃ ipr) ₂ ) 6.60 (1H, d, J = 2.4 Hz) 6.73 (1H, dd, J = 8.3, 2.4 Hz) 7.09 (1H, d, J = 8) .3 Hz)

The above 3-hydroxy-17β- (3,3-
Diphosphono-3-hydroxypropyloxy) -oestra-1,3,5 (10) -triene Tetraisopropyl ester 1.04 g was dissolved in 20 mL of anhydrous chloroform to a solution of trimethylsilyl bromide 1.17 mL under an argon stream under ice cooling. Was added dropwise, and then the mixture was stirred under ice cooling for 1 hour and at room temperature for 20 hours.

To this reaction solution, 2 mL of water was further added under ice cooling, the mixture was stirred at room temperature for 1 hour, the reaction solution was concentrated under reduced pressure, and the residue was azeotropically distilled with ethanol to remove water under reduced pressure. Dried. The obtained powder was washed with benzene, and 3-hydroxy-17β- (3,3-diphosphono-3-hydroxypropyloxy) -estra-1,3,5 (10) -triene (indicated by 29 in the above chemical formula 8). R ⁴ =
(CH ₂ ) ₂ compound) as a pale yellow powder, 670 mg
(Yield 86%) was obtained.

As a result of mass spectrometric analysis of the obtained sample, the mass spectrum (FAB) was m / z: 491 (MH ⁺ ). The NMR spectrum of the sample was as follows.

NMR δ (CD ₃ OD) 0.80 (3H, s), 1.15 to 1.77 (10H, 10H,
m) 1.86 (1H, m), 2.01 to 2.46 (6H,
m) 2.71 to 2.81 (2H, m), 3.48 (1H,
t, J = 8.3 Hz) 3.98 (2H, s-like), 6.48 (1H,
d, J = 1.9 Hz) 6.53 (1H, dd, J = 8.3, 1.9 Hz) 7.06 (1H, d, J = 8.3 Hz)

The solubility of 3-hydroxy-17β- (3,3-diphosphono-3-hydroxypropyloxy) -estra-1,3,5 (10) -triene thus obtained in water is It was very good

Next, examples of the present invention will be described in which the compounds of the present invention as exemplified in the above Production Examples are blended as active ingredients.

[0085]

Example 1 Tablet The ingredients A in Table 1 were mixed, an appropriate amount of demineralized water was added and kneaded, and then vacuum dried at 40 ° C. for 16 hours. The obtained solid is crushed, the component B is added and mixed, and the mixture is tableted to 1000
Individual tablets were produced.

[0086]

[Table 1]

[0087]

Example 2 Capsule The components shown in Table 2 were mixed, and 20 mL of demineralized water was added thereto and kneaded, followed by vacuum drying at 40 ° C. for 16 hours. The obtained solid matter was crushed in a mortar and passed through a 16-mesh sieve to form granules, which were then filled into No. 3 gelatin capsules with a capsule filling machine to produce 1000 capsules.

[0088]

[Table 2]

[0089]

Example 3 Ointment The components shown in Table 3 were mixed by a conventional method to prepare a 1% by weight ointment.

[0090]

[Table 3]

[0091]

Example 4 Injectable Ingredient A in Table 4 was mixed by a conventional method, and distilled water for injection was added to 1000 mL to prepare 0.1 w / v% injectable agent.

[0092]

[Table 4]

<Effect of therapeutic agent according to the present invention> Regarding the therapeutic agent for bone diseases according to the present invention, 3-hydroxy-17β- (3,3-diphosphonopropyloxy) -estra-1, obtained in Production Example 2, was used. 3,5 (10) -triene and estradiol as a comparative example were dissolved in 5% dimethylformamide-95% corn oil at the ratios shown in Table 5, respectively, and Test Examples 1 and 2 and Comparative Examples 1 and 2 were used. A drug efficacy test was conducted.

[0094]

[Table 5] Here, the mol concentrations of the active ingredients in Test Example 1 and Comparative Example 2 are equal.

Ovulation, which is a source of estradiol secretion, was extracted from 7 to 8 8-week-old female Wistar rats to eliminate the influence of estradiol produced in vivo. Nothing was administered to the control group), and to the other groups, each test example and comparative example were subcutaneously administered once a day for 6 weeks from the next day at a ratio of 1 mL per 1 kg body weight. The feeding amount was the same for all rats.

On the day after the final administration of each test example and comparative example, the uterus and left femur of all rats were excised. Regarding the uterus, the weight was measured, the average of each group was calculated, and the data were compared with the standard error. For the left femur, first remove the soft tissue and measure the weight and volume to obtain the specific gravity (hereinafter, this weight is referred to as "wet weight" and the specific gravity is referred to as "wet specific gravity"), and then heat treated at 650 ° C for 18 hours to obtain ash. The specific gravity was calculated by measuring the weight and the volume (hereinafter, this weight is referred to as “ash content weight” and the specific gravity is referred to as “ash specific gravity”), and an average value was calculated for each to compare the data with standard error.

Table 6 shows the uterine weight, the wet weight of the left femur, the wet specific gravity and the ash content weight, and the ash content specific gravity of the control group, the test examples 1 and 2 and the comparative examples 1 and 2 administration groups. In the table, * indicates a risk factor of 5
%, ** indicates a risk rate of less than 1%, and *** indicates a risk rate of less than 0.1%, indicating that there is a significant difference from the control group.

[0098]

[Table 6]

1, 2 and 3, the uterine weight, the ash weight of the left femur and the ash specific gravity of the control group, the test examples 1 and 2 administration groups and the comparative examples 1 and 2 administration groups were compared. As is clear from these results, it was found that the therapeutic agent for bone diseases of the present invention, when used in an appropriate amount, has almost no effect on the uterus and strongly acts on bone to increase bone mass.

[0100]

EFFECTS OF THE INVENTION The therapeutic agent for bone diseases of the present invention is not decomposed in blood and is selectively adsorbed to bone to sufficiently exert a pharmacological effect on bone such as steroid hormones, synthetic estrogens, and plant estrogens. It can be exerted, and it is less likely to accumulate in the uterus, so there is less uterine enlargement and safety is high. Furthermore, if a compound having a hydroxyl group introduced into a part of its structure is used as an active ingredient of the therapeutic agent for bone diseases of the present invention, one having excellent solubility in water can be obtained.

[Brief description of drawings]

1 is a female Wist administered with test examples and comparative examples
The figure which shows the comparison of the uterine weight of ar rat.

FIG. 2 Female Wist administered with test example and comparative example
The figure which shows the comparison of the left femur ash content weight of ar rat.

[Fig. 3] Female Wist administered with test examples and comparative examples
The figure which shows the comparison of the left femur ash specific gravity of ar rat.

[Explanation of symbols]

In the figure, * indicates a risk rate of less than 5%, ** indicates a risk rate of less than 1%,
*** indicates a risk rate of less than 0.1%, which is significantly different from the control group.

Claims (5)

[Claims] 1. A therapeutic agent for bone diseases, which comprises a compound represented by the structural formula of Chemical Formula 1 and / or a pharmaceutically acceptable salt thereof as an active ingredient. [Chemical 1] However, in Chemical formula 1, A-O- represents one kind of residue selected from steroid hormones having human bone resorption inhibitory activity or osteogenesis promoting activity, synthetic estrogen, and plant estrogen, and R ¹ represents a lower alkylene. Group or a lower hydroxyalkylene group, and R ² represents a hydrogen atom or a hydroxyl group, respectively. 2. The therapeutic agent for bone diseases according to claim ^1, wherein R ¹ is a lower hydroxyalkylene group. 3. The therapeutic agent for bone diseases according to claim 1, wherein R ² is a hydroxyl group. 4. The therapeutic agent for bone diseases according to claim 1, wherein the steroid hormone is selected from anabolic steroids, androgens, estrogen, and luteinizing hormone. 5. The steroid hormone is 17β-
It is selected from estradiol, testosterone, norethandrolone, nandrolone and tibolone, and the synthetic estrogen is stilbestrol, monomestrol, hexestrol.
exestrol), Promethestrol,
It is selected from Dienestrol and Benzestrol, and the plant estrogen is Genistein, Biochanin,
Formononetin, Daizo
in), Camertrol (Canmestrol), and Miroestrol (Miroestrol), the therapeutic agent for bone diseases according to claim 1.