Crystalline vitamin D derivative

A crystalline (5Z,7E,23S)-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5,7,10(19)-trie n-1.alpha.,3.beta.,23,25-tetraol monohydrate has an excellent preservation stability, and is therefore useful as a medicine for treating or preventing bone diseases, tumor or psoriasis.

BACKGROUND OF THE INVENTION 
1. Field of the Invention 
The present invention relates to a crystalline vitamin D derivative useful 
for treating or preventing a disease due to abnormality in calcium 
absorption, transportation or metabolism, tumor or psoriasis, a method for 
preparing the derivative and a medical use thereof. Further, the present 
invention relates to a stable pharmaceutical composition containing the 
vitamin D derivative. 
2. Description of the Related Art 
As one of vitamin D derivatives, 
(5Z,7E,23S)-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5,7,10(19)-trie 
n-1.alpha.,3.beta.,23,25-tetraol of formula (2): 
##STR1## 
is known to be useful as a medicine for treating or preventing various 
bone diseases due to abnormality in calcium absorption, transportation or 
metabolism (for example, rickets, osteomalacia and osteoporosis), tumor or 
psoriasis (Japanese Patent KOKAI (Laid Open) No. 63-45249 and U.S. Pat. 
No. 5,030,626). 
Neither Japanese Patent KOKAI (Laid-Open) No. 63-45249 nor U.S. Pat. No. 
5,030,626 specifically disclose a vitamin D derivative hydrate, although 
they specifically disclose the vitamin D derivative in the form of 
non-crystal which is amorphous or non-crystalline powder. 
In the meantime, it is known that an activated vitamin D is topically 
administered in the form of an external formulation to treat intractable 
skin diseases including psoriasis (EP 129003, EP 177920, WO 86/02527 and 
Japanese Patent KOKAI (Laid-Open) No. 63-183534). 
The vitamin D derivative of formula (2) has such problems as poor 
preservation stability and difficult handling in preparation of a 
pharmaceutical formulation. 
In general, an activated vitamin D is chemically unstable, particularly 
under the exposure of light. Thus, when administered in the form of an 
external formulation, the activated vitamin D is particularly unstable in 
the therapeutical use. 
Accordingly, it is important to provide a pharmaceutical composition 
containing an activated vitamin D wherein the vitamin D is stable even 
under the exposure of light after externally administered. 
SUMMARY OF THE INVENTION 
The inventors of the present application have intensively researched to 
solve the problems as stated above and found out that a monohydrate of the 
vitamin D derivative of formula (2) in the form of crystal is extremely 
excellent in preservation stability. Further, the inventors have found out 
a stable pharmaceutical composition containing the vitamin D derivative. 
Thus, the present invention has been completed and accomplished. 
Accordingly, an object of the present invention is to provide a crystalline 
vitamin D derivative of formula (1): 
##STR2## 
Another object of the present invention is to provide a method for 
preparing a crystalline vitamin D derivative of formula (1), which 
comprises the step of: 
subjecting a non-crystalline vitamin D derivative of formula (2): 
##STR3## 
to crystallization treatment in an organic solvent having water added 
thereto. 
Further, another object of the present invention is to provide a 
crystalline vitamin D derivative of formula (1) for therapeutical use. 
Further, another object of the present invention is to provide a medicament 
comprising a crystalline vitamin D derivative of formula (1) as an active 
ingredient. 
Further, another object of the present invention is to provide a medicament 
for the treatment or prophylaxis of a disease due to abnormality in 
calcium absorption, transportation or metabolism, tumor or psoriasis, 
which comprises a crystalline vitamin D derivative of formula (1) as an 
active ingredient. 
Further, another object of the present invention is to provide a medicament 
for the treatment or prophylaxis of psoriasis, which comprises a 
crystalline vitamin D derivative of formula (1) as an active ingredient. 
Further, another object of the present invention is to provide a 
pharmaceutical composition comprising a crystalline vitamin D derivative 
of formula (1) as an active ingredient and a pharmaceutically acceptable 
material. 
Further, another object of the present invention is to provide a method for 
preparing a pharmaceutical composition which comprises the step of mixing 
a crystalline vitamin D derivative of formula (1) with a pharmaceutically 
acceptable material. 
Further, another object of the present invention is to provide use of a 
crystalline vitamin D derivative of formula (1) in the preparation of a 
medicament or pharmaceutical composition for the treatment or prophylaxis 
of a disease due to abnormality in calcium absorption, transportation or 
metabolism, tumor or psoriasis.

DETAILED DESCRIPTION OF THE INVENTION 
The present invention provides the crystalline vitamin D derivative of 
formula (1), that is, crystalline 
(5Z,7E,23S)-26,26,26,27,27,27-hexafluoro-9,10-secocholesta-5,7,10(19)-trie 
n-1.alpha.,3.beta.,23,25-tetraol monohydrate. The crystalline vitamin D 
derivative of formula (1) is extremely excellent in stability, and is 
therefore useful as a medicine for treating or preventing bone diseases, 
tumor or psoriasis. 
The crystalline vitamin D derivative of formula (1) is preferably one 
showing a powder X-ray diffraction spectrum in average values of 
interplanar spacing d and relative intensity as given at Table 1 in 
Example 1 described hereinafter. 
A method for preparing the crystalline vitamin D derivative of formula (1) 
is described below in detail. 
The crystalline vitamin D derivative of formula (1) can be prepared by 
subjecting the non-crystalline vitamin D derivative of formula (2) to 
crystallizion treatment in an organic solvent having water added thereto. 
The organic solvent includes aliphatic hydrocarbons such as n-pentane, 
n-hexane and n-heptane; aromatic hydrocarbons such as benzene and toluene; 
ethers such as diethyl ether, tetrahydrofuran and t-butylmethyl ether; 
ketones such as acetone, methylethyl ketone and methylisobutyl ketone; 
esters such as ethyl acetate; and mixtures thereof. 
The organic solvent is preferably the aliphatic hydrocarbon, aromatic 
hydrocarbon, and mixture of the aliphatic hydrocarbon or aromatic 
hydrocarbon with the ether. 
The crystallization treatment may be carried out by, for instance, 
dissolving the non-crystalline vitamin D derivative of formula (2) in the 
organic solvent at temperature of 10.degree. C. to 35.degree. C.; if 
necessary, distilling off partially the organic solvent; adding water 
thereto; and crystallizing the vitamin D derivative. The vitamin D 
derivative may be crystallized at temperature of 0.degree. C. to 
10.degree. C. 
Preferably, the non-crystalline vitamin D derivative is dissolved in a 
mixture of the aliphatic hydrocarbon or aromatic hydrocarbon with the 
ether, ketone or ester having a lower boiling point than that of the 
aliphatic hydrocarbon or aromatic hydrocarbon, then the ether, ketone or 
ester is distilled off, followed by addition of water to crystallize the 
vitamin D derivative. The aliphatic hydrocarbon or aromatic hydrocarbon 
may also be distilled off in order to raise yield of the crystalline 
vitamin D derivative. 
The amount of the organic solvent after distilled off may be in the range 
of 5 to 200 parts by weight, preferably 8 to 100 parts by weight per part 
by weight of the non-crystalline vitamin D derivative. 
Alternatively, water may be added to the organic solvent, before the 
non-crystalline vitamin D derivative is dissolved in the organic solvent. 
When crystallizing the vitamin D derivative, seed crystals may be added 
thereto. 
Water may be added in an amount of at least 0.1 part by weight, preferably 
1 to 20 parts by weight per part by weight of the non-crystalline vitamin 
D derivative. 
The period of time required for crystallizing the vitamin D derivative may 
vary depending on the crystallization speed, but is usually in the range 
of 30 minutes to 24 hours. 
The thus obtained crystalline vitamin D derivative of formula (1) may be 
recovered by filtration. Upon recovery, the crystalline vitamin D 
derivative may be, if necessary, washed with the organic solvent as used 
in the crystallization treatment. 
The crystalline vitamin D derivative of formula (1) according to the 
present invention may be subjected to dehydration under reduced pressure 
to produce a corresponding crystalline non-hydrate of the vitamin D 
derivative, namely, a crystalline vitamin D derivative of formula (2). 
The crystalline vitamin D derivative of formula (1) exhibits the same 
pharmacological activities as those of the non-crystalline vitamin D 
derivative of formula (2), and is therefore useful as a medicine for 
therapy. More specifically, the crystalline vitamin D derivative of 
formula (1) is useful as a medicine for treating or preventing bone 
disease (for example, rickets, osteomalacia and osteoporosis) caused by 
abnormality in calcium absorption, transportation or metabolism, tumor, or 
psoriasis, likewise the non-crystalline vitamin D derivative of formula 
(2) as described in Japanese Patent KOKAI (Laid-Open) No. 63-45249 and 
U.S. Pat. No. 5,030,626. 
A pharmaceutical composition may be prepared by mixing the crystalline 
vitamin D derivative of formula (1) with a pharmaceutically acceptable 
material. 
The pharmaceutical composition may be in the form of an formulation for 
topical or systemical administration. 
The formulation for topical administration may be an external formulation 
including liquids such as lotions, extracts, suspensions and emulsions; 
and semi-solids such as oleaginous ointments, emulsion ointments and water 
soluble ointments. 
The external formulation is preferably used for treating or preventing 
psoriasis. 
The formulation for systemical administration may be oral preparations in 
the form of tablets, granules, liquids, capsules or soft capsules, or 
other preparations such as injections, suppositories and nasal 
preparations. 
When formulated into an external formulation, the crystalline vitamine D 
derivative of formula (1) may be contained in an amount of 1 ng to 1 mg, 
preferably 50 ng to 50 .mu.g per unit weight of the pharmaceutical 
composition. 
When systemically administered, the crystaline vitamin D derivative of 
formula (1) may be administered at a dose of 2 ng to 100 .mu.g, preferably 
10 ng to 20 .mu.g per human adult per day. 
The formulation may be prepared according to conventionally used methods. 
A pharmaceutically acceptable material for preparing the external 
formulation may usually include bases, preservatives and antioxidants. 
Such bases include hydrocarbon such as mineral oils, solid paraffin, white 
soft paraffin, liquid paraffin, gelled hydrocarbon, dimethylpolysiloxan, 
olieve oil, sesami oil and medium chain triglycerides; aliphatic acid 
esters such as isopropyl myristate, diisopropyl adipate and diethyl 
sebacate; higher alcohols such as stearyl alcohol and cetyl alcohol; water 
soluble polyhydric alcohols such as propylene glycol, polyethylene glycol 
and glycerine; lower alcohols such as ethanol and isopropanol; surface 
active agents such as polyoxyethylene sorbitan fatty acid esters, glycerol 
esters and polyoxyethylene-hardened castor oil; and water. Such 
preservatives include 4-hydroxybenzoic acid esters. Such antioxidants 
include 2,6-di-t-butyl-4-methylphenol and 2-t-butyl-4-methoxyphenol. 
The pharmaceutical composition may preferably have an ultraviolet light 
absorption agent added thereto. The ultraviolet light absorption agent may 
include benzophenones, 4-aminobenzoic acids, cinnamic acids, salicylic 
acids, anthranilic acids, vitamin Es, 
1-(4-methoxyphenyl)-3-(4-t-butylphenyl)propan-1,3-dione, urocanic acids 
and camphor. Such benzophenones include 2-hydroxy-4-methoxybenzophenone, 
2-hydroxy-4-methoxybenzophenone-5-sulphonic acid, sodium 
2-hydroxy-4-methoxybenzophenone-5-sulphonate, 
2,2'-dihydroxy-4,4'-dimethoxybenzophenone, disodium 
2,2'-dihydroxy-4,4'-dimethoxybenzophenone-5,5'-disulphonate, 
2,4-dihydroxybenzophenone and 2,2',4,4'-tetrahydroxybenzophenone. Such 
4-aminobenzoic acids include 4-aminobenzoic acid, ethyl 4-aminobenzoate, 
glycerol 4-aminobenzoate, amyl 4-dimethylaminobenzoate and octyl 
4-dimethylaminobenzoate. Such cinnamic acids include isopropyl 
4-methoxycinnamate, ethyl 4-methoxycinnamate, 2-ethylhexyl 
4-methoxycinnamate, 2-ethoxyethyl 4-methoxycinnamate, potassium 
4-methoxycinnamate and sodium 4-methoxycinnamate. Such salicylic acids 
include octyl salicylate, phenyl salicylate and methyl salicylate. Such 
anthranilic acids include methyl anthranilate and homomentyl 
N-acetyl-anthranilate. Such vitamin Es include natural vitamin E, 
tocopherol acetate and dl-.alpha.-tocopherol. Such urocanic acids include 
urocanic acid and ethyl urocanate. 
Those ultraviolet light absorption agents may be used singly or in 
combination therewith. 
Particularly, 2-ethylhexyl 4-methoxycinnamate and tocopherol acetate are 
preferably used as the ultraviolet light absorption agent. 
The amount of the ultraviolet light absorption agent used may be determined 
depending on the base used in the pharmaceutical composition, but 
preferably in the range of 0.01 to 2% by weight per by weight the 
pharmaceutical composition. 
The present invention will be described in detail below, referring to 
Examples, which are not limitative of the present invention. 
EXAMPLE 1 
Preparation of Crystalline Vitamin D Derivative of Formula (1) 
Ten mg of the non-crystalline vitamin D derivative of formula (2) was 
dissolved into 0.2 ml of diethyl ether, and 1.34 ml of toluene was added 
thereto. The resulting mixture was stirred to obtain a homogeneous 
solution. Then, the solution was concentrated to a half volume under 
reduced pressure, and three drops (about 100 mg) of purified water were 
added thereto. The solution was further stirred at room temperature for at 
least 15 hours. After cooling to about 5.degree. C., the precipitated 
crystals were collected by filtration, whereby 7 mg of the crystalline 
vitamine D derivative of formula (1) was obtained. The thus obtained 
crystalline vitamin D derivative showed a powder X-ray diffraction 
spectrum as given in the following Table 1. 
TABLE 1 
______________________________________ 
Average values of interplanar spacing d and 
relative intensity 
Inter- Inter- 
planar Relative planar Relative 
spacing d intensity spacing d 
intensity 
______________________________________ 
15.77 100 3.92 7 
10.80 10 3.77 10 
9.58 6 3.73 8 
8.37 15 3.67 5 
7.89 13 3.60 7 
7.46 8 3.55 6 
6.65 51 3.46 8 
6.34 31 3.31 15 
5.96 23 3.18 8 
5.79 29 2.96 7 
5.34 54 2.92 6 
4.88 18 2.76 5 
4.78 25 2.70 5 
4.54 29 2.66 6 
4.32 13 2.63 7 
4.23 18 2.54 5 
4.17 13 2.48 5 
______________________________________ 
EXAMPLE 2 
Preparation of Crystalline Vitamin D derivative of Formula (1) 
Ten mg of the non-crystalline vitamin D derivative of formula (2) was 
dissolved into 0.3 ml of diethyl ether, and 0.5 ml of n-heptane was 
further added thereto. The resulting mixture was stirred to obtain a 
homogeneous solution. Then, three drops of purified water were added to 
the solution, and stirred at room temperature for at least 15 hours. The 
precipitated crystals were collected by filtration, whereby 6.5 mg of the 
crystalline vitamin D derivative of formula (1) was obtained. 
EXAMPLE 3 
Preparation of Crystalline Vitamin D Derivative of Formula (1) 
Ten mg of the non-crystalline vitamin D derivative of formula (2) was 
dissolved into 0.3 ml of diethyl ether, and 0.5 ml of n-hexane was further 
added thereto. The resulting mixture was stirred to obtain a homogeneous 
solution. Then, three drops of purified water was added thereto, and 
stirred at room temperature for at least 15 hours. The precipitated 
crystals were collected by filtration, whereby 7 mg of the crystalline 
vitamin D derivatives of formula (1) was obtained. 
EXAMPLE 4 
Preparation of Crystalline Vitamin D Derivative of Formula (1) 
1.56 g of the non-crystalline vitamin D derivative of formula (2) was 
dissolved into 14.4 ml of diethyl ether, and 15.6 ml of toluene was 
further added thereto. The resulting mixture was stirred to obtain a 
homogeneous solution. Then, the solution was concentrated to a half volume 
under reduced pressure. To the solution was added 5.23 ml of purified 
water and 5 mg of seed crystals, then stirred at room temperature for at 
least 15 hours. The precipitated crystals were collected by filtration, 
whereby 1.33 g of the crystalline vitamin D derivative of formula (1) was 
obtained. 
EXAMPLE 5 
Preparation of Crystalline Vitamin D Derivative of Formula (1) 
Ten mg of the non-crystalline vitamin D derivative of formula (2) was 
dissolved into 0.5 ml of acetone, and 1.0 ml of toluene was further added 
thereto. The resulting mixture was stirred to obtain a homogeneous 
solution. Then, the solution was concentrated to a half volume under 
reduced pressure, and three drops of purified water was added thereto, 
followed by stirring at room temperature for at least 15 hours. The 
precipitated crystals were collected by filtration, whereby 7 mg of the 
crystalline vitamin D derivative of formula (1) was obtained. 
EXAMPLE 6 
Preparation of Crystalline Vitamin D Derivative of Formula (1) 
Ten mg of the non-crystalline vitamin D derivative of formula (2) was 
dissolved into 0.2 ml of ethyl acetate, and 1.0 ml of toluene was further 
added thereto. The resulting mixture was stirred to obtain a homogeneous 
solution. Then, the solution was concentrated to a half volume under 
reduced pressure, and three drops of purified water was added thereto, 
followed by stirring at room temperature for at least 15 hours. The 
precipitated crystals were collected by filtration, whereby 5 mg of the 
crystalline vitamin D derivative of formula (1) was obtained. 
EXAMPLE 7 
Preparation of Crystalline Vitamin D Derivative of Formula (1) 
Ten mg of the non-crystalline vitamin D derivative of formula (2) was 
dissolved into 0.2 ml of ethyl acetate, and 1.0 ml of n-hexane was further 
added thereto. The resulting mixture was stirred to obtain a homogeneous 
solution. Then, the solution was concentrated to a half volume under 
reduced pressure, and three drops of purified water was added thereto, 
followed by stirring at room temperature for at least 15 hours. The 
precipitated crystals were collected by filtration, whereby the 
crystalline vitamin D derivative of formula (1) was obtained. 
EXAMPLE 8 
Preservation Stability of Crystalline Vitamin D Derivative of Formula (1) 
and Non-Crystalline Vitamin D Derivative of Formula (2) 
Each of the crystalline vitamin D derivative of formula (1) obtained in 
Example 4 and the non-crystalline vitamin D derivative of formula (2) was 
accurately weighed to obtain the vitamin D derivative at an amount in a 
range of about 1 mg to 1.5 mg. The vitamin D derivatives were preserved in 
atmosphere at room temperature without prevention of light-transmittance, 
for 1 to 5 weeks. 
Thereafter, the preserved vitamin D derivatives were analyzed for the 
remained compound without decomposition according to a absolute 
calibration curve method using a high performance liquid chromatography. 
The obtained results are shown in FIG. 1. The results indicate that the 
crystalline vitamin D derivative of formula (1) almost remained without 
decomposition after the preservation, whereas the non-crystalline vitamin 
D derivative of formula (2) remarkably decomposed. 
EXAMPLE 9 
Preparation of Ointments 
8.27 mg of the crystalline vitamin D derivative of formula (1) obtained in 
Example 4 was mixed with 0.498 g of 2,6-di-t-butyl-4-methylphenol, 1 g of 
2-ethylhexyl 4-methoxycinnamate and 0.2 g of ethanol, followed by addition 
of isopropyl myristate to make the total amount 51 g. The resulting 
mixture was stirred, then fused with 949 g of white soft parrafin at 
40.degree. C. The mixture was cooled to room temperature with stirring to 
obtain 1000 g of ointments. 
EXAMPLE 10 
Preparation of External Liquids 
2.07 mg of the crystalline vitamin D derivative of formula (1) obtained in 
Example 4 was mixed with 0.498 g of 2,6-di-t-butyl-4-methylphenol, 1 g of 
2-ethylhexyl 4-methoxycinnamate and 0.2 g of ethanol, followed by addition 
of isopropyl myristate to make the total amount 51 g. The resulting 
mixture was stirred, and ethanol was added thereto to make the total 
volume 1000 ml. Thus, external liquids were obtained. 
EXAMPLE 11 
Preservation Stability of Pharmaceutical Composition 
The ointments obtained in Example 9 were charged into aluminum tube, then 
preserved at room temperature for one year. 
Thereafter, the ointments were analyzed for the remained crystalline 
vitamin D derivative of formula (1) according to an internal standard 
method using a high performance liquid chromatography. The obtained 
results showed that 100% of the crystalline vitamin D derivative of 
formula (1) remained in the ointments. 
As described in detail hereinbefore, the crystalline vitamin D derivative 
of formula (1) according to the present invention is more excellent in 
preservation stability than the non-crystalline vitamin D derivative of 
formula (2), and can therefore be easily handled in preparing the 
pharmaceutical preparation. The crystalline vitamin D derivative according 
to the present invention can be obtained at high purity by a simple 
crystallization method. 
Furthermore, the pharmaceutical composition according to the present 
invention is excellent in stability. Particularly, the external 
formulation containing an ultraviolet light absorption agent is extremely 
stable, and can preserve the vitamin D derivative without decomposition 
for a long period of time enough for therapeutical use, even under the 
exposure of light of 1000 Lux.