JPH0651624B2 - Drugs for prevention of bone loss or treatment or prevention of regenerative disorders - Google Patents

Description

The present invention relates to the prevention of bone loss or the treatment of regenerative disorders, which comprises a vitamin D ₂ series 1α, 25-dihydroxylated compound and a pharmaceutically acceptable carrier. Or, it relates to a preventive drug.

More specifically, the present invention provides a pharmaceutically acceptable carrier for 1α, 25-dihydroxyvitamin D ₂ and its (24R) -epimer, the corresponding 5,6-trans-isomer, and certain C-25-alkyl derivatives. The present invention relates to a drug for preventing bone loss or treating or preventing regenerative disorders.

(Problems to be Solved by Prior Art and Invention) The importance of the hydroxylated form of vitamin D as a substance for controlling the metabolism of calcium and phosphate in animals and humans is as follows.
To date, it has been well recognized through numerous disclosures in patents and general literature, and as a result of these, hydroxyvitamin D
Derivatives are finding increasing clinical and veterinary use as agents for the treatment and treatment of bone disorders associated with disorders of calcium metabolism. Vitamin D ₃ is known to be hydroxylated in vivo to 25-hydroxyvitamin D ₃ and then to 1α, 25-dihydroxyvitamin D ₃ ,
Here, the latter is generally accepted as the active hormone form of vitamin D ₃ . Similarly, a very effective vitamin D ₂ metabolite, 1α, 25-dihydroxyvitamin D ₂ (1α, 25- (OH) ₂ D ₂ ) is a vitamin D ₂ to 25-
It is formed through the hydroxy vitamin _{D 2 25-OH-D 2} ). Both of these hydroxylated vitamin D ₂ compounds have been isolated and identified (Deluca et al., US Pat. No. 3,585,22).
No. 1, 3,880, 894). These metabolites derived from vitamin D ₂ is (S) of carbon 24 - characterized by the stereochemistry. The object of the present invention is to provide a drug for preventing bone loss or treating or preventing regenerative dyscrasia, which comprises a vitamin D ₂ 1α, 25-dihydroxylated compound and a pharmaceutically acceptable carrier.

(Means for Solving Problems) A drug for preventing or ameliorating bone loss comprising a vitamin D ₂ 1α, 25-dihydroxylated compound and a pharmaceutically acceptable carrier has been developed. It was Here, the vitamin D ₂ -based 1α, 25-dihydroxylated compound is a compound having the general structures A and B shown below.

(In the formula, R ₁ , R ₂ and R ₃ are selected from the group consisting of hydrogen and an acyl group, and X is an alkyl or aryl group. In these structures, the absent center of carbon 24 is (R) or ( S) may have a sequence.) Of the vitamin D ₂ -based 1α, 25-dihydroxylated compounds, R ₁ , R ₂ and R ₃ in the general structure A or B are hydrogen,
A compound in which X is an alkyl group having 1 to 6 carbon atoms (however,
A 5,6-cis compound of the above general structure A, wherein C-5
Except compounds where the 4-methyl substituent has the S-sequence and X is a methyl group. ) Is preferable as an active ingredient of the drug of the present invention.

That is, the present invention is: 1. a compound selected from the following group (In the formula, R ₁ , R ₂ and R ₃ are hydrogen, and X is a carbon number 1
~ 6 alkyl groups. However, the above-mentioned 5,6-cis compound except that the C-24-methyl substituent has an S-sequence and X is a methyl group is excluded. 2. A drug for preventing bone wear or treating or preventing regenerative disorders, which comprises at least one of the above) and a pharmaceutically acceptable carrier; 2. The bone according to the above 1, wherein X in the compound of the above formula is a methyl group. 3. A preventive or regenerative agent for the treatment or prophylaxis of ataxia, 3. A compound according to the above formula, wherein the C-24-defective center has an (R) -sequence. 4. A drug for treating or preventing disorder, 4. Prevention of bone loss or treatment or prevention of regenerative disorder according to claim 1, wherein the C-24-defective center of the compound of the above formula has (S) -sequence. 5. A drug for the prevention or treatment of bone loss or regenerative dysfunction according to claim 1, wherein the compound of the above formula is 1α, 25-dihydroxy-24-epivitamin D _2. compound l [alpha], 25-dihydroxy-5,6-trans - the vitamin D ₂ as set forth in claim 1, wherein the bone Therapeutic or prophylactic agents Worn preventing or reproduction disorders diseases, 7. the formula of the compound is l [alpha], 25-dihydroxy-5,6-trans-24-epi - a vitamin D ₂ of the first claim of the bone Drugs for preventing wear or treating or preventing regenerative disorders, 8.1 α, 25-dihydroxy-5,6-trans-vitamin D ₂
And / or 1α, 25-dihydroxy-5,6-trans-24
- epi - said containing vitamin D ₂ according to claim 1 wherein the prevention or reproduction disorders diseases for the treatment or prevention agents wear bone, 9.1α, 25- dihydroxy-5,6-trans - vitamin D ₂
And 1α, 25-dihydroxy-vitamin D ₂ for preventing bone loss or treating or preventing regenerative dystonia according to claim 1, 10.1α, 25-dihydroxy-5,6-trans-vitamin D
_{2. A} drug for the prevention of bone loss or the treatment or prophylaxis of regenerative dysfunction, which comprises ₂ and 1α, 25-dihydroxy-5,6-trans-24-epivitamin D ₂ , and 11.1α, 25-dihydroxy Vitamin D ₂ , 1α, 25-dihydroxy-24-epi-Vitamin D ₂ , 1α, 25-dihydroxy-5,6-trans-vitamin D ₂ and 1α, 25-dihydroxy-5,6-trans-24-epi- A drug for preventing bone loss or treating or preventing regenerative dyscrasia according to claim 1, which contains vitamin D _2. 12.1α, 25-dihydroxyvitamin D ₂ and 1α, 25-dihydroxy-24-epi-vitamin D The first consisting of ₂
13. A drug for the prevention of bone loss or the treatment or prevention of regeneration disorder according to the item 13.1α, 25-dihydroxy-5,6-trans-vitamin D
₂ and 1α, 25-dihydroxy-5,6-trans-24-epi-vitamin D ₂ or 1α, 25-dihydroxy-24-
The first containing either one of epi-vitamin D ₂
The drug according to item 1, wherein the drug is a drug for preventing or treating bone loss or for treating or treating regenerative disorders, 14. The drug according to item 1, wherein the drug is a drug for preventing or treating osteoporosis.

Specific examples of the vitamin D ₂ -based 1α, 25-dihydroxylated compound include 1α, 25-dihydroxyvitamin D ₂ , the corresponding (24R) -epimer, 1α, 25-dihydroxy-24-epivitamin D ₂ , respectively. 5,6-trans-isomers, namely 5,6-trans-1α, 25-dihydroxyvitamin D ₂ and 5,6-trans-1α, 25-dihydroxy-24-epivitamin D ₂ , and C-25 of the compound of
Included are alkyl or aryl homologs, ie, compounds where X above is an ethyl, propyl, isopropyl or phenyl group.

The term "acyl" as used herein means an aliphatic acyl group having 1 to 6 carbon atoms (alkanoyl group) including all possible isomeric forms such as formyl, acetyl, butyryl, isobutyryl and valeryl, and an aromatic acyl group (aroyl group). , For example, vinzoyl or a methyl, halo or nitro substituted benzoyl group or the general formula ROOC (CH ₂ ) _n CO- or ROOCCH ₂ -O-CH ₂ CO
Means an acyl group derived from a dicarboxylic acid having-(wherein n is an integer (including 0 and 4) having a value of 0 to 4 and R is hydrogen or an alkyl group). Representative of such dicarboxylic acyl groups are oxalyl,
Malonyl, succinoyl, glutaryl, adipyl and diglycolyl. The term "alkyl" includes hydrocarbon groups of 1 to 6 carbons, including all isomeric forms, for example:
It means methyl, ethyl, propyl, isopropyl, butyl, isobutyl and the like. The term "aryl" refers to phenyl, benzyl or alkyl substituted phenyl group isomers.

Specific examples of chemical processes for obtaining the compounds according to the invention are depicted in the attached Process Scheme I. In the following description of this process, numbers (eg, 1, 2, 3, etc.) indicate particular products so numbered in Process Scheme I. The wavy line for the substituent (methyl) in C-24 indicates that this substituent is R or S.
It indicates that any of the sequences may be taken.

A preferred starting material for the process for obtaining the compounds according to the invention is the vitamin D-ketal derivative of structure (1). In general, compound (1) is used as a mixture of 24R and S epimers (for example, when both C-24 epimers of a 1α, 25-dihydroxyvitamin D ₂ compound are required) and individual 24R and S epimers are used. Separation of the epimers is conveniently performed at a later stage of the method. However, the net of (1)
The 24S or pure 24R epimers are equally suitable as starting materials, whereby the former compound is processed by the indicated synthetic steps to provide the (24S) -1α, 25-dihydroxy product and the latter compound is Similar treatment yields the corresponding (24R) -1α, 25-dihydroxylated product.

Starting material (1) is converted to the desired 1α-hydroxylated form via a cyclovitamin D derivative (Deluca et al., US Pat. Nos. 4,195,027 and 4,260,549). Thus, conventional treatment of compound (1) with toluenesulfonyl chloride gives the corresponding C-3-tosyl compound (2), which is subjected to solvolysis in alcoholic medium to give the new 3,5- This produces a cyclovitamin D derivative (3). Structure (3) in solvolysis in methanol
Yields a cyclovitamin with Z = methyl. However, the use of other alcohols such as ethanol, 2-propanol, butanol and the like in this reaction gives analogous cyclovitamin D compounds (3) where Z is an alkyl group derived from the alcohol, eg ethyl, isopropyl, butyl and the like. . When the intermediate (3) was subjected to allyl oxidation with selenium dioxide and hydroperoxide, 1α of the structure (4) was obtained.
Yields the hydroxy-analog. Compound (4) is subsequently acetylated to give ₁ -of the structure (5, R ₁ = acetyl)
Give acetate. If desired, other 1-0-acylated compounds (structure 5, where R ₁ = acyl such as formate, propionate, butyrate, benzoate, etc.)
Is prepared by a similar conventional acylation reaction. This 1-O-acylated derivative is then subjected to acid catalyzed solvolysis. When this solvolysis is carried out in a solvent containing water, the 5,6-cis-vitamin D intermediate of the structure (6, R ₁ = acyl, R ₂ = H) and the corresponding 5,6-trans-compound (structure 7, R ₁ = acyl, R ₂ = H) is obtained in a ratio of about 3-4: 1. These 5,6-cis and 5,6-trans-isomers can be separated at this stage, for example by high performance liquid chromatography. If desired, the C-1-O-acyl group can be removed by basic hydrolysis and R ₁ and R ₂
Compounds (6) and (7) having ═H can be obtained. Also, if desired, these 1-O-acylates can be further acylated at the C-3-hydroxy group using conventional acylation conditions to provide structure (6) or (7) (wherein R ₁ and R _{2 s} may be the same as or different from each other, and the corresponding 1,3-di-O-acylated product of (representing an acyl group) can be obtained. Alternatively, the hydroxycyclovitamin of structure (4) is subjected to acid catalyzed solvolysis in a medium containing a low molecular weight organic acid to give structures (6) and (7) (where R ₁ = H, R ₂ = acyl). Where the acyl group is derived from the acid used in the solvolysis reaction) to obtain the 5,6-cis and trans compounds. The next step in the process is to remove the ketal protecting group and produce the corresponding 25-ketone. Conversion of ketals to ketones occurs under the acidic conditions required for ketal hydrolysis of 22 (23) -2 double bonds.
This step is very important as the isomerization to the 23 (24) -conjugated position must be accomplished in unison. Furthermore, the conditions must be chosen to avoid the elimination of labile allyl C-1-oxygen functional groups. This conversion can be successfully accomplished by careful hydrolysis at moderate temperatures with organic acid catalysis. Thus, the 5,6-cis-compound (6) is treated with p-toluenesulfonic acid in aqueous alcohol to give the corresponding ketone (8). In order to avoid unintended removal of the C-1 oxygen functional group during this reaction, the C-1 of compound (6)
Protecting the hydroxy group (eg R ₁ = acyl, R ₂
= Hydrogen or acyl) is preferred.

Subsequent reaction of the ketone (8) with a methyl Grignard reagent gives the desired 1α, 25-dihydroxyvitamin D ₂ compound of structure (9). If the starting material used in the above method is a mixture of two C-24-epimers, compound (9) will contain 24S and R-epimers (9a and 9a, respectively).
It will be obtained with a mixture of b). Separation of this epimeric mixture can be accomplished by chromatographic methods, including 1α, 25-dihydroxyvitamin D ₂ (structure 9a, 24S-
Stereochemistry) and its 24R-epimer, 1α, 25-dihydroxy-24-epivitamin D _{2 of} structure 9b, are both obtained in pure form. Such separation of epimers is of course not necessary if the compounds are used as a mixture.

When the 5,6-trans-25-ketal intermediate of structure (7) was subjected to ketal hydrolysis in a similar manner, structure (10)
5,6-transketone intermediate of 5-6-trans-1α, 25-dihydroxyvitamin D ₂ of Grignard reaction structure (11) with a reagent of methylmagnesium bromide or the like was added to this process. Depending on the nature of the starting material (1) used, it is provided as the 24S or 24R-epimer or as a mixture of both epimers. If obtained as a mixture of epimers, the epimers are separated by chromatography to give 5,6-trans-1α, 25-dihydroxyvitamin D ₂ (11a) and its 24R-epimer,
5,6-trans-1α, 25-dihydroxy-24-epitavitamin D ₂ of structural formula (11b) is obtained. The reaction steps with these 5,6-trans-intermediates can be carried out in a manner quite similar to that applicable to the 5,6-cis compounds described above.

The structures (8) and (10) of the novel side chain ketones have various 1
It is the most useful and versatile intermediate in that it can be used to prepare α, 25-dihydroxyvitamin D ₂ side chain analogs. In particular, these keto-intermediates are 5,6-cis- or 5,6-trans- having the side chain general formula:
It can be useful in the preparation of 1α, 25-dihydroxyvitamin D ₂ analogs.

(Where X is an alkyl or aryl group) For example, treatment of ketone (8) with ethylmagnesium bromide provides a corresponding hydroxyvitamin D ₂ analog wherein X is an ethyl group in the side chain general formula shown above. give. Similarly, treatment of (8) with isopropylmagnesium or phenylmagnesium bromide provides side chain analogs where X is isopropyl or phenyl, respectively. Treatment of the 5,6-trans-25-ketone intermediate of structure (10) with an alkyl or aryl Grignard reagent in a similar manner has a side chain which is an alkyl or aryl group derived from the Grignard reagent used for X 5 It gives the 6,6-trans-vitamin D ₂ analogue.

Isotopically labeled Grignard reagents of keto intermediates (8) or (10) (eg C ³ H ₃ -MgBr, ¹⁴ CH ₃ MgBr,
C ² H ₃ MgBr, etc.) to give 1α, 25-dihydroxyvitamin D ₂ or its trans isomer and the corresponding C
In -24- epimeric isotopically-labeled form, that is, X of the side chain shown above is _{^{C 3 H 3, 14 CH 3}} , C 2 H 3, 13 CH 3 , or other isotope-labeled with alkyl It is also clear that it provides a convenient way of preparing the compound, which in turn is selected from aryl groups.

The alkyl or aryl analogues of 5,6-cis or trans-1α, 25-dihydroxy-vitamin D ₂ described above are effective substituents of the parent compound when very high lipophilicity is required, On the other hand, the above-mentioned isotope-labeled compound can find use as a reagent for analytical application.

Furthermore, for therapeutic applications, the free hydroxy compounds represented by structures A and B above, wherein R ₁ , R ₂ and R are
₃ = H) is commonly used, but in some such applications the corresponding hydroxy-protected derivative would be effective and preferred. Such hydroxy-protected derivatives are represented, for example, by the general formulas A and B above, R ₁ , R ₂
And one or more of R ₃ is an acylated compound showing an acyl group.

Such an acyl derivative can be obtained by subjecting a free hydroxy compound to a conventional acylation method such as hydroxyvitamin D.
It can be conveniently prepared by treating either of the _two products with an acyl halide or acid anhydride in a suitable solvent such as pyridine or an alkylpyridine. By proper choice of reaction time, acylating agent, temperature and solvent, partially or fully acylated acylated derivatives of structure A or B as known in the art can be obtained. For example, 1α, 25-
Treatment of dihydroxyvitamin D ₂ (9a) with acetic anhydride in a pyridine solvent at room temperature gives 1,3-diacetate, while the same reaction at elevated temperature yields the corresponding 1,3,25-
This produces triacetate. This 1,3-diacetate can be further acylated at the C-25 position with different acyl groups. Treated with, for example, benzoyl chloride or succinic anhydride,
The 1,3-diacetyl-25-benzoyl- or 1,3-diacetyl-25-succinoyl-derivatives are obtained, respectively. 1,3,25
The triacyl derivative can be subjected to selective hydrolysis in mild base to provide the 1,3-dihydroxy-25-O-acyl compound. The free hydroxy groups here can be reacylated with different acyl groups if desired. Similarly, 1,3-diacyl derivatives can be subjected to partial acyl hydrolysis to give 1-O-acyl and 3-O-acyl compounds, which can be further reacylated with different acyl groups. Other hydroxyvitamin D ₂ products (eg 9b, 11a / b or their corresponding 25
-Alkyl or aryl analogs) provides the corresponding desired acyl derivatives of structure A or B (wherein any or all of R ₁ , R ₂ and R ₃ are acyl).

1α, 25 as well as the previously known vitamin D ₂ metabolite
-Dihydroxyvitamin D ₂ (9a), the novel compounds according to the invention show a marked vitamin D-like activity and thus a desirable alternative to known vitamin D ₂ or D _{3 in} a wide range of therapeutic or veterinary applications. It becomes a thing. Particularly preferred products in this respect are structures 9b and 11a and 11b or their acylated derivatives. This novel compound is well known in various diseases such as rickets of vitamin D resistance, osteomalacia, hypoparathyroidism, skeletal dysplasia, pseudohypoparathyroidism, osteoporosis, Paget's disease and medical practice. It can be used to ameliorate or correct various calcium and phosphorus imbalances that occur as a result of similar bone and mineral related disease states. The compounds can also be used for the treatment of animal mineral imbalances such as fever in lactation, foot frailty in poultry or improved eggshells in chickens. Their use for the treatment of osteoporosis is of particular noteworthy.

During menopause, women suffer significant bone loss, ultimately leading to bone deficiency disease, which eventually results in vertebral compression, natural fractures and long bone fractures. This disease, commonly known as post-menopausal osteoporosis, has become an important medical problem in the United States and in other countries where women live for at least 60-70 years.
In general, the disease is often diagnosed by squeezed fractures of one or more vertebrae with X-ray evidence of bone loss, often accompanied by bone pain and reduced physical activity. It is known that the disease is accompanied by diminished calcium absorption capacity, diminished levels of sex hormones, especially estrogens and androgens, and a negative calcium balance.

The methods of treating this disease have changed significantly. For example,
Supplementing calcium itself has not been successful in preventing or treating the disease. Injection of sex hormones, especially estrogen, which has been reported to be effective in preventing the rapid wear and tear of bone experienced in postmenopausal women, is difficult because of its fear of carcinogenicity. It was With respect to other treatment methods, various results have been reported again, and some of them may combine high dose vitamin D, calcium and fluoride. The main problem with this approach is that fluoride is structurally unfavorable bone,
It is known to induce so-called focal bones and, in addition to this, increase the incidence of fractures and produce many side effects of causing a gastrointestinal reaction by administering a large amount of fluoride.

Similar symptoms are present in senile osteoporosis and steroid-induced osteoporosis, the latter of which is recognized to result from glucocorticoid (corticosteroid) treatment for long-term disease states.

Various vitamin D ₃ metabolites increase calcium uptake and maintenance in the body of mammals showing evidence of bone loss or having such physiological tendencies, but in response to physiological demand. It is also characterized by its complementary vitamin D-like properties that in turn mobilize calcium in bone. Epi compounds of the compounds according to the present invention, especially 24-
Epi-1α, 25-dihydroxyvitamin D ₂ (24-epi-
1,25- (OH) ₂ D ₂ ) is outstandingly suitable for the prevention or treatment of physiological disorders in mammals characterized by bone loss. Because they exhibit some of the perceived Vitamin D-like properties that affect calcium metabolism, such as increasing intestinal calcium transport and acting on bone mineralization, but even at high doses, serum calcium levels are high. Is not increased. This observed property makes it clear that administration of this compound does not fluidize the bone. This fact, together with the ability of this compound to administer bone mineralization when administered, is useful in the prevention or treatment of the prevailing calcium disorders evidenced by bone loss, such as post-menopausal osteoporosis, senile osteoporosis, and steroid-induced osteoporosis. On the other hand, it is shown to be an ideal compound. This compound can be readily applied for the prevention or treatment of other disease states in which bone loss is an indicator, for example for the treatment of patients undergoing renal dialysis which faces bone loss as a result of dialysis. It's obvious.

Examples The following examples contribute to their outstanding suitability for the prevention or treatment of disease states exhibiting bone loss 24-Epi-1,25 (OH)
It will help explain the properties of ₂ D ₂ .

Example 1 Weaning male rats into a journal of Suda et al.
Journal of Nutrition 100 , 1
049-1052 (1970) described in the vitamin D deficient diet,
They were placed under special diet conditions modified to contain 0.02% calcium and 0.3% phosphorus. After 2 weeks on this special diet, the animals were given 1,25-dihydroxyvitamin D ₂ or 24-ep
The 1,25-dihydroxyvitamin D _2, 5% ethanol solution 0.1m propanediol were given daily subcutaneous injections.
Twelve hours after the last dose, animals were sacrificed and blood calcium and intestinal calcium transport was measured. These measurement results
The indicated levels of compound administered are shown in FIGS. 1 and 2. The measurement of intestinal calcium transport shown in Figure 2 was performed by the American Journal of Martin and Deluca.
Physiology (American Journal of Physio-logy) 21
6 , 1351-1359 (1969).

Example 2 Weanling male rats were treated with high calcium (1.2% calcium), low phosphorus (0.1%) as described by Suda et al., Supra.
% Phosphorus) special dietary conditions. Rats were fed this special diet for 3 weeks and were divided into two groups. One group was given 1,25 (OH) ₂ D ₂ and the other was given 24-EP 1,25 (OH) ₂ D ₂ , both groups containing 5% ethanol in propanediol. The compound was given subcutaneously in 0.1 m at the dose level of the compound indicated by the data points in FIG. After continuing this dosing daily for 7 days, the animals were sacrificed and the amount of serum inorganic phosphorus was measured. Results are shown in FIG.

The femur of the rat was taken out and the ash content of the bone was measured. The femur was cut free of attached connective tissue and extracted in anhydrous alcohol for 24 hours and in diethyl ether for 24 hours using a Soxhlet extractor. The bone was ashed at 600 ° F for 24 hours. Ash weight was determined by measuring constant weight. Results are shown in FIG.

The results of the acute toxicity test are shown in Table 1 below. As shown in Table 1, the semi-lethal dose of 24-epi-1α, 25- (OH) ₂ D ₂ in mice was 1.25 to 2.5 mg / kg by oral administration,
The semi-lethal dose in rats is over 5.0 mg / kg for oral administration, over 0.2 mg / kg for subcutaneous administration, 5.0 mg / kg for males and 10.0 mg / kg for females by intraperitoneal administration. It was a larger value. This is, for example, 1α, 25
-It is about the same value as in the case of dihydroxyvitamin D ₃ (1α, 25- (OH) ₂ D ₃ ). Moreover, 24-epi-1α, 2 is more preferable than 1α-hydroxyvitamin D ₃ (1α- (OH) D ₃ ).
5- (OH) ₂ D ₂ is less toxic.

The results of the two studies described in Examples 1 and 2 above are:
24-Epi-1,25- (OH) ₂ D ₂ is 1α, 25-dihydroxyvitamin D ₃ (1,25- (OH) ₂ D in response to the occurrence of bone mineralization and stimulation of intestinal calcium transport. It is explained that it is as effective as ₂ ). In short, there is no real meaningful difference between the two groups of FIG. 2 and FIG. On the other hand, in the case of a low phosphorus diet, the increase in serum inorganic phosphate resulting from fluidization of bone is very significantly affected by 1,25- (OH) ₂ D ₂ , but 24-epi-1,25- ( It means that it is hardly stimulated by (OH) ₂ D ₂ . Similarly, extremely high doses of serum calcium levels of about 750 pmol / day (first
The 24-epi compound did not show any effect on the mobilization of calcium from the bone, as even pointed out in the figure). On the other hand, 1,25-dihydroxyvitamin D
₂ , the fluidization effect is clear even at much lower doses. Elevated serum calcium in rats on a low-calcium diet is a measure of the potential for bone mobilization, and increased blood phosphorus in low-phosphorus diets is also a measure of bone mobilization. , These results are 24-epi-1,25
-(OH) ₂ D ₂ is able to completely stimulate its unexpected properties, namely intestinal calcium transport and new bone mineralization, but with minimal efficacy in mobilizing bone calcium,
It exhibits properties which make this compound particularly suitable for the treatment of disease states which are manifested by bone loss.

24-epi -1,25- (OH) ₂ D _2, unique properties as described above, the method and extent which could not have been realized so far, various vitamin D response process (intestinal calcium absorption, bone It provides a rare opportunity to control mineral fluidization and bone mineralization). This possibility is that the epi-compounds of the invention are administered to mammals alone (with suitable and acceptable excipients) or in combination with other vitamin D derivatives which exhibit a full spectrum of D-like activity. Arises from the fact that By such means, therefore, 24-epi-
It makes it possible to combine the active properties of analogues with the general activity of other vitamin D metabolites or analogues. Administration of 24-epi-1,25- (OH) ₂ D ₂ alone, as indicated above,
It stimulates intestinal calcium transport and bone mineralization, but no or minimal bone mineral mobilization occurs. But,
The latter activity is known for vitamin D derivatives (eg 1,25-
(OH) ₂ -D ₃ , 1α, 25- (OH) ₂ D ₂ , 1α-OH-D ₃ and related analogues) can be induced by co-administration of one or more of them. By controlling the relative amounts of compounds administered, a degree of control over intestinal calcium absorption versus the relative strength of the bone mineral mobilization process can be exerted in ways not previously possible with vitamin D derivatives. . The bone mobilization activity of co-administration of 24-epitaxy compounds with other vitamin D compounds is particularly advantageous in situations where some bone mobilization is required. For example, in some circumstances, bone must first be fluidized before new bone is sacrificed. Under such conditions, vitamin D or vitamin D derivatives that induce bone mobilization, such as 1α-hydroxyvitamin D ₃ or D ₂ , 1α, 25-dihydroxyvitamin D ₃ or D ₂ , 25-
Hydroxyvitamin D ₃ or D ₂ , 24,24-difluoro-2
5-hydroxyvitamin D ₃ , 24,24-difluoro-1α,
25-dihydroxy - vitamin D _3, 24- fluoro-25
Hydroxyvitamin D ₃ , 24-Fluoro-1α, 25-dihydroxyvitamin D ₃ , 2β-Fluoro-1α-hydroxyvitamin D ₃ , 2β-Fluoro-25-hydroxyvitamin D ₃ , 2β-Fluoro-1α, 25-dihydroxy-
Vitamin D ₃ , 26,26,26,27,27,27-hexafluoro-1
α, 25-dihydroxy-vitamin D ₃ , 26,26,26,27,27,2
7-hexafluoro-25-hydroxyvitamin D ₃ , 24,2
5-dihydroxyvitamin-D ₃ , 1α, 24,25-trihydroxyvitamin D ₃ , 25,26-dihydroxyvitamin-D
₃ , 1α, 25,26-trihydroxyvitamin D _{3 in combination} with 24-epi-1,25- (OH) ₂ D ₂ to control the ratio of 24-epi compound and bone mobilizing vitamin D compound Curing allows the degree of fluidization of bone to be adjusted until the desired medical or physiological purpose is achieved. Suitable and effective mixtures are, for example, 1α, 25-dihydroxyvitamin D ₂ and 1
α, 25-Dihydroxy-24-epivitamin D ₃ (9a and 9b)
Or a mixture of the corresponding 5,6-trans compounds (11a and 11b), or these free hydroxy compounds or other combinations of the four compounds as their acylated forms. The compounds of the present invention or their conjugates with other vitamin D derivatives or other therapeutic agents are orally administered as a sterile parenteral solution by injection or infusion, easily administered through the skin or from the digestive tract in the form of suppositories. To be done. Advantageously, the compound is given in a dose of 0.1-100 μg per day. About 0.5-25 μg / day for osteoporosis
Doses of are generally effective. This compound is administered alone or in combination with other vitamin D derivatives, the proportion of each compound in combination depending on the particular disease state targeted and the desired degree of bone mineralization and / or bone fluidization. .
The actual amount of 24-epi compound used in the treatment of osteoporosis, where the preferred compound is 24-epi-1α, 25- (OH) ₂ D ₂ , is not critical. In all cases, the compound should be used in an amount sufficient to induce bone mineralization. Using more than about 25 μg of the compound in combination with 24-epi-compound or bone fluidization-derivatized vitamin D derivative per day is generally unnecessary to achieve the desired results and is economical. It is not a proper implementation. In practice, higher doses of the compound are used when the goal is to treat the disease state, while lower doses are generally used for prophylactic purposes. However, in any case, the dosage given, as is well known to those skilled in the art, will depend on the particular compound being administered, the disease to be treated, the condition of the patient and other appropriate medical practice, activity of the drug. It should be understood that the patient's response will be adjusted by anything that requires modification.

The agents of the present invention can be prepared by combining the compounds of the present invention with non-toxic pharmaceutically acceptable carriers.
The carrier includes a usual carrier as well as a shaping agent. Such carriers may be solid or liquid such as corn starch, lactose, sucrose, peanut oil, olive oil, sesame oil and propylene glycol. If a solid carrier is used, the dosage form of the agents of the invention may be tablets, capsules, powders, troches, or lozenges. If a liquid carrier is used, soft gelatin capsules, syrups or suspensions,
The emulsion or solution can be a dosage form. The dosage form may also contain preservatives, stabilizers, adjuvants such as wetting or emulsifying agents, dissolution promoters and the like. They may also contain therapeutically valuable substances such as other vitamins, salts, sugars, proteins, hormones or other pharmaceutical compounds.

The compounds according to the present invention are explained in more detail by reference examples 1 to 7 below. In these reference examples, specific products represented by Arabic numerals (for example, compound 1,
(2,3, etc.) refer to such numbered structures in Process Scheme I.

Reference Example 1 1α-hydroxy-3,5-cyclovitamin D (4, Z =
Methyl) A solution of compound (1) (50 mg) (as a mixture of 24R and S epimer) in dry pyridine (300 μ) was treated with 50 mg of p-toluenesulfonyl chloride at 4 ° C. for 30 hours. The mixture was poured onto ice / saturated NaHCO ₃ with stirring and the product was extracted with benzene. The combined organic layers are NaHCO ₃ aqueous solution,
It was washed with H ₂ O, aqueous CuSO ₄ solution and water, dried over MgSO ₄ and evaporated. The crude 3-tosyl derivative (2) was stirred with anhydrous methanol (10 m) and NaHCO ₃ (150 mg) at 55 ° C. for 8.5.
Heated for an hour and subjected to solvolysis. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to ~ 2m. Benzene (80m) is then added, the organic layer is washed with water, dried,
Evaporated. The resulting cyclovitamin (3, Z = methyl) can be used for the next oxidation without further purification.

Crude product (3) in CH ₂ Cl ₂ (4.5 m) was ice-cooled, SeO ₂
(5.05 mg) and t-BuOOH (16.5μ) were added to a CH ₂ Cl ₂ (8m) solution containing anhydrous pyridine (50μ). 0
After stirring for 15 minutes at ° C, the reaction mixture was warmed to room temperature. further
After 30 minutes, transfer the mixture to a separatory funnel and add 10% NaOH.
It was shaken at (30m). Add ether (150m),
The separated organic phase was washed with 10% NaOH, water, dried and evaporated. The oily residue was applied to a silica gel thin layer plate (20 x 20
cm plate, AcOEt / hexane 4: 6), 1α-
20 mg of the hydroxy derivative (4, Z = methyl) was obtained. Mass spectrum m / e: 470 (M ⁺ , 5), 438 (20), 87 (100); NMR (CDCl
₃ ) δ0.53 (3H, s, 18-H ₃ ), 0.63 (1H, m, 3-H), 4.19 (1H, d, J = 9.
5Hz, 6-H), 4.2 (1H, m, 1-H), 4.95 (1H, d, J = 9.5Hz, 7-H), 5.17
And 5.25 (2H. Each m, 19-H ₂ ), 5.35 (2H, m, 22-H and 23-H). Reference Example 2 Acetylation of compound (4) Cyclovitamin (4, Z = methyl) (18mg ) In pyridine (1 m) and acetic anhydride (0.33 m) was heated at 55 ° C. for 2 hours. The mixture was poured into ice cold saturated NaHCO ₃ and extracted with benzene and ether. Combine organic extracts with water, saturated
Wash with aqueous CuSO ₄ and NaHCO ₃ , dry and evaporate 1-
An acetoxy derivative (5, Z = methyl, acyl = acetyl) (19 mg) was obtained. Mass spectrum m / e: 512 (M, 5), 42
0 (5), 87 (100); NMR (CDCl ₃ ) δ 0.53 (3H, s, 18-H ₃ ), 4.18 (1
H, d, J = 9.5Hz, 6-H), 4.97 (2H, m, 7-H and 19-H), 5.24 (2H, m, 1-
H and 19-H), 5.35 (2H, m, 22-H and 23-H). Reference Example 3 1α-acetoxy-3,5-cyclovitamin (5) (R ₁ =
Acetyl) sorborisis Cyclovitamin (5) (4.5 mg) dioxane / H ₂ O
The solution in a 3: 1 mixture (1.5m) was heated at 55 ° C. p-Toluenesulfonic acid (1 mg in 20 [mu] of water) was then added and heating continued for 15 minutes. The mixture was poured into saturated Nacho ₃ / ice, benzene and ether extracted. The organic phase is NaHCO ₃
Washed with water and dried over MgSO ₄ . Evaporation of the solvent gave a residue containing compounds (6) (where R ₁ = acetyl, R ₂ = H) and (7) (where R ₁ = acetyl, R ₂ = H), which , 2% in hexane as eluent 2-
HPLC with propanol (6.2 mm x 25 cm Zorbax-Sil)
Separated by above chromatography. If needed,
The product can be further purified by rechromatography.

Reference Example 4 Ketone (8) is obtained by ketal hydrolysis of compound (6).

To a solution of ketal (6, R ₁ = acetyl, R ₂ = H) (1.35 mg) in ethanol (1.5 m) was added p-toluenesulfonic acid (0.34 mg in 45 μm water) and the mixture was refluxed.
Heated for 30 minutes. The reaction mixture was poured into diluted NaHCO ₃ and extracted with benzene and ether. The organic extracts are combined, washed with water, dried over MgSO _4, and evaporated. High pressure liquid chromatography of the crude mixture (4% 2-
Propanol / hexane, 6.2mm × 25cm Zorbax-Sil)
Contains some unreacted ketal (6) (0.12 mg, collected at 48 m) and the desired ketone (8, R ₁ = acetyl and R ₂ = H) (0.36 mg, 52 m) characterized by the following data:
Gathered in). Mass spectrum m / e: 454 (M ⁺ ,
9), 394 (17), 376 (10) 134 (23), 43 (100); NMR (CDCl ₃ ) δ 0.5
3 (3H, s, 18-H ₃ ), 1.03 (3H, d, J = 6.5Hz, 21-H ₃ ), 1.13 (3Hd, J =
7.0Hz, 28-H ₃ ), 2.03 (3H, s, CH ₃ COO), 2.12 (3H, s, CH ₃ CO), 4.
19 (1H, m, 3-H), 5.03 (1H, m, 19-H), 5.33 (3H, broad m, 19
H, 22-H and 23-H), 5.49 (1H, m, 1-H), 5.93 (1H, d, J = 11Hz, 7-
H), 6.37 (1H, d, J = 11Hz, 6-H); UV (EtOH) λmax 266nm, 250n
m, λmin 255 nm. Reference example 5 Obtaining products (9a) and (9b) by reacting ketone (8) with methylmagnesium bromide Excess in ketone (8, R ₁ = acetyl, R ₂ = H) in anhydrous ether CH ₃ MgBr
(2.85M solution in ether). The reaction mixture was stirred at room temperature for 30 minutes, then quenched with aqueous NH ₄ Cl and extracted with benzene, ether and CH ₂ Cl ₂ . The organic phase was washed with dilute NaHCO ₃ , dried over MgSO ₄ and evaporated. The mixture of (9a) and (9b) thus obtained was subjected to high performance liquid chromatography (6% 2-propanol / hexane, 4.6 mm x 25 cm Zorbax-Sil) to elute in order of pure epimer (9a) and (9a). 9b) was obtained. 1α, 25-dihydroxyvitamin D ₂ (9a): UV (EtOH) λmax 265.5nm, λ
min277.5nm; mass spectrum m / e: 428 (M ⁺ , 6), 410 (4), 35
2 (4), 287 (6), 269 (10), 251 (10), 152 (42), 134 (100), 59 (9
9); NMR (CDCl ₃ ) δ 0.56 (3H, s, 18-H ₃ ), 1.01 (3H, d, J = 6.5H
z, 28-H), 1.04 (3H, d, J = 6.5Hz, 21-H), 1.14 and 1.18 (6H, each s,
26-H and 27-H), 4.24 (1H, m, 3-H), 4.43 (1H, m, 1-H), 5.01 (1H,
m, 19-H), ~ 5.34 (3H, Broad m, 19-H, 22-H and 23-H), 6.02 (1H,
d, J = 11Hz, 7-H), 6.39 (1H, d, J = 11Hz, 6-H). 1α, 25-dihydroxy-24-epivitamin D ₂ (9b): UV (Et
OH) λmax265.5nm, λmin277.5nm; mass spectrum, m / e
428 (M ⁺ , 13), 410 (9), 352 (7), 287 (11), 269 (15), 251 (13), 1
52 (52), 134 (100), 59 (97). Reference Example 6 Conversion of compound (7) to 5,6-trans-1α, 25-dihydroxyvitamin D ₂ compounds (11a) and (11b) Ketal intermediate Hydrolysis of the form (1, R ₁ = acetyl, R ₂ = H) using the conditions described in Reference Example 2 yielded the corresponding
The structure of 5,6-trans-25-ketone (10, R ₁ = acetyl, R ₂ = H) is provided, and this ketone is then prepared in Reference Example 3
Reaction with methylmagnesium bromide using conditions similar to that gives epimers (11a) and (11b) which are separated by high performance liquid chromatography (HPLC) to give pure 1α, 25-dihydroxy-. 5,6-Trans-vitamin D ₂ (11a) and 1α, 25-dihydroxy-5,6-
Trans-24-epivitamin D ₂ (11b) can be obtained. If required, the structural designation can be confirmed by isomerization to the respective 5,6-cis compound (9a, 9b) by known techniques.

5,6-trans-1α, 25-dihydroxyvitamin D ₂ (11a): U
V (EtOH) λmax273.5nm, λmin230nm; mass spectrum, m / e 428 (M,
8), 410 (3), 287 (3), 269 (7), 251 (34), 134 (100), 59 (78) .5,6-trans-1α, 25-dihydroxy-24-epivitamin D ₂
(11b): UV (EtOH) λmax273.5nm, λmin230nm; mass spectrum, m
/ e 428 (M ⁺ , 10), 410 (4), 352 (4), 287 (5), 269 (9), 251 (8), 15
2 (37), 134 (100), 59 (82). Reference Example 7 Preparation of alkyl and aryl analogs of 1α, 25-dihydroxyvitamin D ₂ compound Ketone intermediate (8) (R ₁ = acetyl, R ₂ = H) is treated with (a) ethyl magnesium bromide, (b) propyl magnesium bromide, (c) isopropyl magnesium bromide, (d) butyl magnesium bromide, and (e) phenyl magnesium bromide, using conditions similar to those described in Reference Example 5. Then, the corresponding hydroxyvitamin D _{2 represented} by the following formula
Is obtained.

In the above formula, X is as follows.

(A) Ethyl (b) Propyl (c) Isopropyl (d) Butyl (e) Phenyl 5,6-trans-ketone intermediate (10) (R ₁ = acetyl,
R ₂ = H) is similarly treated with the Grignard reagents listed above to give the corresponding 5,6-trans-hydroxyvitamin D ₂ product having the formula shown below.

In the above formula, X is as follows.

(A) Ethyl (b) Propyl (c) Isopropyl (d) Butyl (e) Phenyl A suitable starting material for the compounds according to the invention is the vitamin D-ketal derivative of structure (1), which is a British patent specification. It can be obtained according to Process Chimes II and III described in US Pat. No. 2,127,023 or US Pat. No. 4,448,721.
In general, it is convenient to use compound (1) as a mixture of 24R and 24S epimers (for example when both C-24 epimers are desired) and to separate the individual 24R and 24S epimers later. However, the pure 24S- or pure 24R-epimer of (1) is likewise suitable, the former providing the 24S-1α, 25-dihydroxy product and the latter the corresponding 24-R-product. Process scheme I Process scheme II Process scheme III

[Brief description of drawings]

Figure 1 shows 1,25- (OH) ₂ D ₂ or 24-epi-1,25- (OH)
A graph showing the relationship between the dose of ₂ D ₂ compound and the amount of calcium in serum, FIG. 2 shows 1,25- (OH) ₂ D ₂ or 24-epi-1,25- (OH)
A graph showing the relationship between the dose of ₂ D ₂ compound and the amount of intestinal calcium transport, FIG. 3 shows 1,25- (OH) ₂ D ₂ or 24-epi-1,25- (OH)
Fig. 4 is a graph showing the relationship between the dose of ₂ D ₂ compound and serum inorganic phosphorus. Fig. 4 shows 1,25- (OH) ₂ D ₂ or 24-epi-1,25- (OH)
₂ is a graph showing the relationship between ₂ D ₂ compound dose and total bone ash content.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Rafal R. Inventor. Sishinski Poland Pastura 1 Warsaw 02-093 Department of Chemistry University of Warsaw (72) Inventor Yoko Tanaka United States 53711 Wisconsin Madison Hirtoptop Drive 613 (56) References US Patent 4260549 (US, A)

Claims (14)

[Claims] 1. A compound selected from the following group: (In the formula, R ₁ , R ₂ and R ₃ are hydrogen, and X is a carbon number 1
~ 6 alkyl groups. However, the above-mentioned 5,6-cis compound except that the C-24-methyl substituent has an S-sequence and X is a methyl group is excluded. A pharmaceutical agent for preventing bone wear or treating or preventing regenerative disorders, which comprises at least one of the above) and a pharmaceutically acceptable carrier. 2. A drug for the prevention of bone loss or the treatment or prevention of regenerative disorders as claimed in claim 1, wherein X in the compound of the above formula is a methyl group. 3. A drug for the prevention or the treatment or prevention of remodeling disorders of bone according to claim 1, wherein the asymmetric center of C-24 of the compound of the above formula has (R) -sequence. 4. A drug for the prevention or treatment of bone atrophy or regenerative dyscrasia according to claim 1, wherein the asymmetric center of C-24 of the compound of the above formula has (S) -sequence. 5. A drug for the prevention of bone loss or for the treatment or prophylaxis of regenerative dyscrasia according to claim 1, wherein the compound of the above formula is 1α, 25-dihydroxy-24-epivitamin D ₂ . 6. The use according to claim 1, wherein the compound of the above formula is 1α, 25-dihydroxy-5,6-trans-vitamin D ₂ , for the prevention of bone loss or the treatment or prevention of regenerative disorders. Drug. 7. A method according to claim 1, wherein the compound of the above formula is 1α, 25-dihydroxy-5,6-trans-24-epivitamin D ₂ , preventing bone loss or treating regenerative disorders. Or a preventive drug. 8. A 1α, 25-dihydroxy-5,6-trans-
Vitamin D ₂ and / or 1α, 25-dihydroxy-5,6-
The drug for preventing bone loss or treating or preventing regenerative dyscrasia according to claim 1, which contains trans-24-epi-vitamin D ₂ . 9. 1α, 25-dihydroxy-5,6-trans-
Vitamin D ₂ and 1α, 25-dihydroxy-vitamin D ₂
A drug for preventing bone wear or treating or preventing regenerative dyscrasia according to claim 1. 10. The method according to claim 1, which comprises 1α, 25-dihydroxy-5,6-trans-vitamin D ₂ and 1α, 25-dihydroxy-5,6-trans-24-epi-vitamin D _2. For the prevention of bone loss or the treatment or prevention of regenerative disorders. 11. A 1α, 25-dihydroxyvitamin D ₂ , 1
α, 25-dihydroxy-24-epi-vitamin D ₂ , 1α, 2
5-dihydroxy-5,6-trans-vitamin D ₂ and 1
The drug for preventing bone wear or for treating or preventing regenerative dysfunction according to claim 1, which contains α, 25-dihydroxy-5,6-trans-24-epi-vitamin D ₂ . 12. Prevention of bone loss or treatment of regenerative dyscrasia according to claim 1, which comprises 1α, 25-dihydroxyvitamin D ₂ and 1α, 25-dihydroxy-24-epi-vitamin D _2. Prophylactic drug. 13. 1α, 25-Dihydroxy-5,6-trans-vitamin D ₂ and 1α, 25-dihydroxy-5,6-trans-24-epi-vitamin D ₂ or 1α, 25-dihydroxy-24- The drug for preventing bone loss or treating or preventing regenerative dyscrasia according to claim 1, which contains one of epi-vitamin D ₂ . 14. The drug according to claim 1, which is a drug for preventing or treating osteoporosis.