NO175429B - Analogous Process for Preparing Therapeutically Active 16-Dehydro-Vitamin D3 Derivatives - Google Patents

Description

The invention relates to an analogue method for the preparation of therapeutically active compounds with the formula

wherein R is hydrogen or hydroxy, and A is -C=C-,

-CH=CH- with E configuration or -CH2-CH2-.

These can be used in pharmaceutical preparations such as

comprises one, two or more compounds of the formula I, for the treatment of hyperproliferative skin diseases, such as psoriasis, and for the treatment of neoplastic diseases, such as leukemia.

Examples of C 1 -alkyl groups to which reference is made

below, are methyl, ethyl, propyl, isopropyl, butyl and t-butyl. Examples of aryl-C 1 -alkyl groups are benzyl, phenethyl and phenylpropyl. Examples of aryl groups are phenyl and p-tolyl.

Halogen can be bromine, chlorine, fluorine or iodine.

Compounds of the formula I prepared according to the invention

are compounds A to F as defined below:

A: 1,25-dihydroxy-16-dehydrocholecalciferol;

B: 25-hydroxy-16-dehydrocholecalciferol;

C: 1,25-dihydroxy-16,23E-bisdehydrocholecalciferol;

D: 25-hydroxy-16,2 3E-bisdehydrocholecalciferol;

E: 1,25-dihydroxy-16-dehydro-23-didehydrocholecalciferol; and

F: 25-hydroxy-16-dehydro-23-didehydrocholecalciferol;

among which the 1,25-dihydroxylated compounds A, C and E are preferred.

The compounds with the formulas Ia and Ib (included by the formula I) are prepared according to the invention as described in schemes 1, 2 and 3 by reacting a corresponding compound with the formula I which instead of the two or three hydroxy groups contains two or three protected hydroxy groups with the formula •

wherein R x and R 3 are C 1 -alkyl and R 2 is C 1 -alkyl, aryl

or aryl-C 1 -alkyl,

with an agent capable of removing the protecting groups.

wherein A is as described above and R 1 and R 3 are independently <C>1-4 alkyl and R 2 is independently <C>1-4 alkyl, aryl, or aryl-C 1-4 alkyl. wherein R 1 , R 2 and R 3 are as described above.

wherein R 1 , R 2 and R 3 are as described above, X is chlorine, bromine or iodine and Ts is tosyl.

The intermediates of formula II, including those of formulas IIa, IIb and IIc are new.

In scheme 1, the compound with the formula li~~b is formed into a compound with the formulas IVa or IVb by reaction with the corresponding compound with the formula

where Ph is phenyl; R 4 is H or -OS 1 , R 2 , R 3 ), and R 1 R 2 and R 3 are as described above.

The reaction is carried out at -60 to -90°C, preferably -75°C, in a polar, aprotic, organic solvent, e.g. dry ether or preferably dry tetrahydrofuran (THF), in the presence of a strong base, such as an alkyllithium, e.g. butyllithium.

The protecting groups of a compound of formulas IVa or IVb are removed by reaction with a fluorine salt, e.g. tetrabutylammonium fluoride in a polar organic solvent, e.g. ether or preferably THF, to give a corresponding compound of formula Ia or Ib.

In scheme 2, the compound of formula V is oxidized to the compound of formula VI by treatment with an oxidizing agent, e.g. 2,2'-bipyridinium chlorochromate or preferably pyridinium chlorochromate in an aprotic organic solvent such as methylene chloride.

The compound with the formula VI is converted into a compound with the formula IIb, by reaction with e.g. a (trialkylsilyl)-imidazole such as (trimethylsilyl)imidazole, in an aprotic organic solvent such as THF, or preferably methylene chloride.

The compound with the formula V can also be hydrogenated to the compound with the formula VII in particular by reaction with a reducing agent, e.g. lithium aluminum hydride, preferably in the presence of an alkali metal alkoxide, e.g. sodium methoxide, in an aprotic organic solvent, e.g. ether, or preferably THF, at a reflux temperature (about 68°C for THF), for 10 - 20 hours.

The resulting compound of formula VII is oxidized to the compound of formula VIII by treatment with an oxidizing agent as described above for the oxidation of V to VI.

The compound of formula VIII is converted to a compound of formula IIa, by reaction with a (trialkylsilyl)imidazole, as described above for the conversion of VI to IIb.

In scheme 3, the compound of formula X is reacted in ether, preferably THF, at reflux temperature with magnesium. The resulting Grignard solution is treated with cuprous iodide and then the compound of formula IX is added.

The resulting compound of formula XI is reacted with a fluoride salt, e.g. tetrabutylammonium fluoride in ether or preferably THF.

The obtained compound of formula XII can be oxidized as described above for the oxidation of V to VI.

The resulting compound of formula XIII is converted to a compound of formula 11c by reaction with a (trialkylsilyl)imidazole as described above for the conversion of VI to IIb.

To prepare a compound of formula IX, the compound of formula

(Tetrahedron 40, 1984, 2283) is reacted with a tosylating agent, such as a p-toluenesulfonyl halide, e.g. the chloride, in an organic base, e.g. collidine or preferably pyridine. The resulting compound of formula is then converted to a compound of formula IX by reaction with a trialkylsilyl chloride, e.g. trimethylsilyl chloride, in the presence of imidazole and in an aprotic organic solvent, e.g. THF or methylene chloride. To prepare a compound of the formula X, a compound of the formula can be converted into a compound of the formula

wherein X is as above, by conversion with a methyl Grignard reagent such as methylmagnesium bromide in ether. The compound of formula XVII is converted to a compound of formula X, by reaction with a trialkylsilyl chloride, as described above

for conversion of XV to IX.

To prepare the compound of formula V, the compound of formula XV above is reacted with a cyanide-forming agent, e.g. sodium cyanide, in an aprotic organic solvent, e.g. dimethyl sulfoxide (DMSO), at a temperature between .80 and 100°C for 1 to 5 hours to give a compound of the formula

This is converted into the compound with the formula by reaction with a reducing agent, e.g. diisobutylaluminum hydride, followed by hydrolysis with e.g. a mineral acid, such as hydrochloric acid. The reduction is carried out in an aprotic organic solvent, e.g. methylene chloride, at -10 to 10°C for 20 to 90 min. The compound of formula XIX is converted to the compound of formula

by reaction with a mixture of triphenylphosphine, carbon tetrabromide and zinc dust, in an aprotic organic solvent, e.g. methylene chloride, for 1 to 30 hours.

The compound with the formula XX is converted to the compound with the formula

when trading with a strong base, e.g. butyllithium, in a polar aprotic solvent, e.g. THF, at -80° to -70°C, for 1 to 3 hours. The compound of formula XXI is converted to the compound of formula by reaction with (trimethylsilyl)imidazole in an aprotic organic solvent, e.g. THF or methylene chloride. This compound is converted into the compound with the formula

when trading with a strong base, e.g. butyllithium and then with acetone. The reaction is carried out in an aprotic organic solvent, e.g. THF at -80 to -60°C. The compound of formula XXIII is deprotected to give the compound of formula V (in Scheme 2) by reaction with a fluorine salt, e.g. tetrabutylammonium fluoride in an organic

solvent, e.g. ether or THF.

The compound with formula I stimulates differentiation and reduces the proliferation of human keratinocytes. Accordingly, they are useful as agents in the treatment of hyperproliferative skin diseases, such as psoriasis, basal cell carcinoma, disorders or keratinization and keratosis. The compound of formula I is also useful as an agent in the treatment of neoplastic diseases, such as leukemia.

The activity of compounds of the formula I as agents for the treatment of hyperproliferative skin diseases can be demonstrated e.g. by test procedures known in the art, as set forth in The Society for Investigative Dermatology 1986, 709-714. The effects of compounds A to F above on the morphological differentiation of cultured human keratinocytes compared to the effect of 1,25-dihydroxycholecalciferol (compound X) are shown in Tables 1 to 4 below, as the number (xl0<A>) of human keratinocytes in the culture. A compound which induces the differentiation of basal cells into squamous and enveloping cells is useful as an agent in the treatment of skin diseases characterized by disturbances of keratinization, such as psoriasis.

The activity of compounds of formula I as agents for the treatment of hyperproliferative skin diseases can also be demonstrated by determining the number of human keratinocytes cultured and the number of enveloping cells formed, as well as the number of squamous carcinoma cell lines (SCC-115) cultured in cultures, in the presence of said compounds. The results are given in tables 4 and 5:

The above results show that the compounds of formula I induce the differentiation of skin cells and are therefore useful in the treatment of hyperproliferative disorders of the skin, such as psoriasis.

To demonstrate the activity of the compounds of formula I as agents for the treatment of neoplastic diseases, the anti-proliferative (AP) and differentiation-inducing (DI) effects of compounds A to F and human promyelocytic HL-60 tumor cells were evaluated. Table 6 gives the AP effect in percentage reduction of the cell number and in the concentration ID50 of the compound that reduces the number of cells by 50%. The DI effect is expressed as the percentage of differentiated cells and as the concentration ED50 of the compounds that induces a 50% differentiation of the cells.

These data indicate that each of the subject compounds inhibits the proliferation of human promyelocytic cells, in vitro, although they are not toxic to the cells. Moreover, the cells differentiate towards a more mature phenotype at the same doses that inhibit proliferation. From these results it can be seen that all the compounds tested are useful as an agent in the treatment of neoplastic diseases, such as leukemia.

The compounds of formula I can be administered orally for the treatment of neoplastic diseases or for the treatment of hyperproliferative skin diseases, to warm-blooded animals in need of such treatment, e.g. to an adult human, in doses that lie in the range between about 0.1 and icT/ug per day and night.

For the treatment of hyperproliferative skin diseases, compounds of formula I can also be administered locally to warm-blooded animals in need of such treatment, in doses of about 1 to 1000 µg per gram of local preparation per day and night.

Oral dosage forms comprising compounds of formula I can be incorporated e.g. in capsules or tablets with pharmaceutically acceptable carriers. Examples of such carrier materials that can be incorporated into the capsules are binders such as gum tragacanth or gelatin; excipients such as dicalcium phosphate; disintegrating agents, such as corn starch; lubricants such as magnesium stearate; sweeteners such as sucrose; flavoring agents such as peppermint. The tablets can be coated with shellac, sugar or both. A syrup or elixir may contain a sweetener, methyl and propyl parabens as preservatives, a coloring agent and a flavoring agent.

Topical dosage forms comprising compounds of formula I include ointments and creams comprising preparations having an oily, adsorbable, water soluble and emulsion type base, such as lanolin and polyethylene glycols. Local dosage forms also include gels, lotions, powders and aerosols. The local preparations can also be used in the treatment of inflammations in the skin, or in mucous membranes, e.g. the mucous membranes of the mouth or lower part of the colon.

Lotions, i.e. liquid preparations varying from simple solutions to aqueous or water/alcoholic preparations containing finely divided substances, may contain suspending or dispersing agents, such as cellulose derivatives, e.g. ethyl or methyl cellulose; gelatin or gum, which incorporates the active ingredient in a carrier consisting of water, alcohol or glycerin. Gels are semi-solid preparations, made by gelling a solution or suspension of the active ingredient in a carrier. The carriers, which may be aqueous or non-aqueous, are gelled using a gelling agent, e.g. carboxypolymethylene, and is neutralized to a real gel consistency using alkalis, e.g. sodium hydroxide, or amines, such as polyethylene cocoamine.

Example 1

a) A mixture of 3.24 g of [1 (R*), 3aR*- (3a/3, 4a, 7aa) ] - 3a, 4, 5, 6, 7 , 7a-hexahydro-4-hydroxy-/ 3,7α-dimethyl-3H-inden-1-ethanol, 30 ml of pyridine and 3.51 g of p-toluenesulfonyl chloride are stirred at 0° for 18 hours. After addition of ice and dilution with water, the mixture is extracted with methylene chloride. The organic phase is washed with IN H 2 SO 4 , saturated NaHCO 3 , then dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:1:5) to give 4.6 g (82%) of [l(R<*>),3aR<*->

(3a/3,4a,7aac)]-3a,4,5,6,7,7a-hexahydro-4-hydroxy-/3,7a-dimethyl-3H-indene-1-ethanol-4-methylbenzenesulfonate, [a ]W + 31.9°

(c 0.53, CHCl 3 ).

b) To a solution of 4.61 g of the product from a) in 22 ml of DMSO, 1.10 g of sodium cyanide is added, and the mixture is heated at

90°C for 2 hours. After cooling to room temperature, the mixture is pumped to remove the solvent, and then diluted with water. The mixture is extracted with ether. The organic phase is washed with saturated salt solution, dried and evaporated. The residue is chromatographed on silica gel with methylene chloride-hexane-ethyl acetate (86:7:7) to give 2.52 g (91%) of [ 1 (R*), 3aR*- (3a/3, 4a, 7aa) ] - 3a,4,5,6,7,7a-hexahydro-4-hydroxy-(3,7a-dimethyl-3H-indene-l-propanenitrile, [a] V + 29.2° (c 0.65, CHCl3) .

c) To a mixture of 6.85 ml diisobutylaluminum hydride in hexane and 5.2 ml methylene chloride at -6°C is added a solution of

0.430 g of the product from b) in 10 ml of methylene chloride. The mixture is stirred at -6°C for 55 min. After adding saturated ammonium chloride, the mixture is hydrolysed with 3N HCl ether (2:1). The aqueous layer is extracted with ether. The organic layers are washed with saturated salt solution, dried and evaporated.

The residue is chromatographed on silica gel with ethyl acetate-hexane (1:2) to give 260 mg (60%) [ 1 (R*), 3aR*-(3a/3, 4a, 7aa) ] - 3a, 4,5,6 ,7,7a-hexahydro-4-hydroxy-[3,7a-dimethyl-3H-inden-l-propanal, [a]<2>D<2><+> 43.1° (c 0.32, CHC13 ).

d) A mixture of 1.77 g of triphenylphosphine, 2.23 g of carbon tetrabromide, 441 mg of Zn dust and 23 ml of methylene chloride is stirred at 25°C

for 31 hours. To this mixture is added a solution of 0.430 g of the product from c) in 38 ml of methylene chloride, and the mixture is stirred for 18 hours. The mixture is diluted with pentane and insoluble material is filtered off. The insoluble fraction is dissolved in methylene chloride, and the solution is diluted again with pentane. After filtration, the combined filtrates are evaporated. The residue is purified on silica gel with 1:4 ethyl acetate-hexane to give 0.490 g (67%) of [1(R*),3aR*-(3a/3,4a,7aa)]-1-(4,4-dibromo -1-methyl-3-butenyl)-3a,4,5,6,7,7a-hexahydro-7a-methyl-3H-inden-4-ol, [a]<2>D<2> + 14.4 ° (c 0.55, CHCl 3 ). e) To a solution of 0.680 g of the product from d) in 31 ml of THF at -75°C, 3.77 ml of a 1.6M solution of butyllithium in hexane is added dropwise. The mixture is stirred at -75°C for 1 hour, and at 25°C for 1 hour. After addition of saturated salt solution, the mixture is diluted with saturated aqueous NaHCO 3 and extracted with ether. The organic phase is washed with saturated salt solution, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:4) to give 0.350 g (89%) of [ 1 (R*), 3aR*-(3aj3,4a,7aa)]-3a,4,5,6, 7, 7α-Hexahydro-7α-methyl-1-(1-methyl-3-butynyl)-3H-inden-4-ol, [α]2D2 + 30.7° (c 0.42, CHCl3). f) To a solution of 1.29 g of the product from e) in 80 ml of methylene chloride, 3.59 g of 1-(trimethylsilyl)imidazole is added.

The mixture is stirred at 25°C for 3 hours. After the addition of

40 ml of water and stirring for 20 min., the mixture is extracted with ethyl acetate. The organic phase is washed with water and saturated salt solution, dried and evaporated. The residue is purified on silica gel with ethyl acetate-hexane (1:15) to give 1.70 g (99%) of [1(R*), 3aR*-(3aØ,4a,7aa)]-3a,4,5, 6,7,7a-hexahydro-7a-methyl-1-(1-methyl-3-butynyl)-4-[(trimethylsilyl)oxy]-3H-indene,

[α]<2>D° + 39.7° (c 0.30, CHCl 3 ).

g) To a solution of 1.70 g of the product from f) in 48 ml of THF at -75°C, 6.01 ml of 1.6 M butyllithium in

hexane. After stirring for 40 min. 3.05 ml of acetone is added, and the mixture is stirred at -75°C for 20 min. and at 25°C for 75 min. After addition of 40 ml of a 1:1 mixture of 2M KHCO 3 and 1M potassium sodium tartrate, the mixture is stirred for 20 min. and then extracted with ethyl acetate. The organic phase is washed with saturated salt solution, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:5) to give 1.62 g (89%) of [1(R*), 3aR*-(3aj8,4a,7aa)]-6-(3a,4,5 ,6,7,7α-hexahydro-7α-methyl-4-[(trimethylsilyl)oxy]-3H-inden-1-yl)-2-methyl-3-heptyn-1-ol, [a]<2>D ° + 39.7 ° (c 0.30, CHCl 3 ). h) To a solution of 1.62 g of the product from g) in 53 ml of THF, 15.5 ml of 1M tetrabutylammonium fluoride in THF is added.

The mixture is stirred for 50 min. After dilution with half-saturated NaHCO 3 , the mixture is evaporated to remove most of the solvent, and extracted with ethyl acetate. The organic phase is washed with half-saturated salt solution, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:1) to give 1.17 g (82%) of [ 1 (R*), 3aR*-(3a/3, 4a, 7aa) ] - 3a,4, 5,6,7,7α-hexahydro-1-(1,5-dimethyl-5-hydroxy-3-hexynyl)-7α-methyl-3H-inden-4-ol, m.p. 105-107°C.

i) To a solution of 0.720 g of the product from h) in 44 ml of methylene chloride, 1.59 g of sodium acetate and 3.18 g of 2,2'-bipyridinium chlorochromate are added. The mixture is stirred for 2 hours. An additional 1.59 g of 2,2'-bipyridinium chlorochromate is then added and stirring is continued for 2 hours. Then, after adding 6 ml of 2-propanol, the mixture is diluted with water and extracted with ether-ethyl acetate (1:1). The organic phase is washed with water, IN H 2 SO 4 , saturated NaHCO 3 and saturated saline. After drying, the solution is evaporated, and the residue

chromatographed on silica gel with ethyl acetate-hexane (1:1) to give 0.560 g (78%) of [ 1 (R*), 3aR*-(3a/J,7aa) ]-3,3a,5,6,7 ,7a-hexahydro-1-(5-hydroxy-1,5-dimethyl-3-hexynyl)-7a-methyl-4H-inden-4-one, [a]<2>D° +35.3° (c 0.36, CHCl 3 ).

j) To a solution of 0.552 g of the product from i) in 70 ml of methylene chloride, 2.00 g of 1-(trimethylsilyl)imidazole is added. After stirring for 17 hours and adding 22 ml of water, the mixture is extracted with ethyl acetate. The organic phase is washed with water and saturated salt solution, then dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:4) to give 0.693 g (99%) of [1(R*), 3aR*-(3a/3,7aa)]-3,3a,5,6, 7,7α-hexahydro-1-(1,5-dimethyl-5-[(trimethylsilyl)-oxy]-3-hexynyl)-7α-methyl-4H-inden-4-one, [ct]2D° + 29, 5° (c 0.20, CHCl 3 ).

k) To a solution of 2.00 g of [3S-(1Z,3a,5/3)]-[2-[3,5-bis[[(1,1-dimethyl)dimethylsilyl]oxy]-2- methylenecyclohexylidene]ethyl]diphenylphosphine oxide in 45 ml of THF at -75°C, 1.87 ml of 1.6M butyllithium in hexane is added dropwise.

After stirring for 6 min. a solution of 0.693 g of the product from j) in 26 ml of THF is added dropwise. After stirring at -75°C for 70 min. and adding a 1:1 mixture of 1M potassium sodium tartrate and 2M KHCO 3 , the mixture is extracted with ethyl acetate. The organic phase is washed with saturated salt solution, dried and evaporated. The residue was chromatographed on silica gel with ethyl acetate-hexane (1:15) to give 1.23 g (87%) of (let,3)3,5Z,7E)-1,3-bis[ [1,1-dimethylethyl) dimethylsilyl ] oxy-25-[(trimethylsiliyl)oxy]-9,10-secocholesta-5,7,10(19) ,16-tetraen-3-yne, [ct]2D3 <+> 47.1° (c 0 ,21, CHC13) .

1) To a solution of 0.228 g of the product from k) in 11 ml of THF, 1.92 ml of 1M tetrabutylammonium fluoride in THF is added. The mixture is stirred for 16 hours. After dilution with water, the mixture is extracted with ethyl acetate. The organic phase is washed with semi-saturated salt solution and saturated salt solution, dried and evaporated. The residue is purified on silica gel with ethyl acetate-hexane (3:1) to give 0.126 g (96%) of 1,25-dihydroxy-16-dehydro-23-didehydrocholecalciferol, [-a] V + 21.5°

(c 0.20, MeOH).

Example 2

a) As described in example lk), but starting from 0.343 g of [5S-(1Z)]-[2-[5-[[(1,1-dimethylethyl)dimethylsilyl]-oxy]-2-methylenecyclohexylidene]ethyl ]diphenylphosphine oxide and 0.186 g of the product from Example 1j), 0.205 g (80%) of (3/3,5Z,7E)-3-[[ (1,1-dimethylethyl)dimethylsilyl]oxy] -25-[(trimethylsilyl)oxy]-9,10-secocholesta-5,7,10(19),16-tetraen-23-yne, MS m/e 580 (M*). b) ' By treating 0.248 g of the product from a) as described in example 11), 0.153 g (91%) of 25-hydroxy-16-dehydro-23-didehydrocholecalciferol was obtained, [ajV +99.6°

(c 0.25), MeOH).

Example 3

a) To a mixture of 0.146 g of lithium aluminum hydride, 0.211 g of sodium methoxide and 6.5 ml of THF are added dropwise at

0°C to a solution of 0.180 g of the product from example lh) in 13 ml of THF. The mixture is heated at 68°C for 16 hours, and

cooled back to 0°C. After dilution with 13 ml of ether and addition of 0.30 ml of water and 0.26 ml of 10% aqueous NaOH, the mixture is stirred at room temperature for 1 hour and filtered. The solids are rubbed out with ether and filtered off. The combined filtrates are evaporated and chromatographed on silica gel with ethyl acetate-hexane (1:2) to give 0.179 g (99%) of [1(R*),1(3E),3a/3,4a,7acr)]-( 3a,4,5,6,7,7a-hexahydro-1-(5-hydroxy-1,5-dimethyl-3-hexenyl)-7a-methyl-1H-inden-4-ol, [a]V <+ >11.5° (c 0.33, CHCl 3 ).

b) To a solution of 0.120 g of the product from a) in 10 ml of methylene chloride, add 0.500 g of pyridinium dichromate and 25 mg

pyridinium p-toluenesulfonate. The mixture is stirred for 135 min. After adding 40 ml of ether, the mixture is stirred for 5 min.

and filtered. The solids are rubbed out with ether and filtered off. The combined filtrates are washed with saturated aqueous CuSOA, water, half-saturated aqueous NaHCO 3 and brine. The organic phase is dried and evaporated. The residue is chromatographed on silica gel with 35% ethyl acetate-hexane to give 90 mg (76%) of [1 (R *) , 1 (3E) , (3aj8, 7aa) ] - 3, 3a, 5, 6,7,7a-hexahydro-1-(5-hydroxy-1,5-dimethyl-3-hexenyl)-7a-methyl -4H-inden-4-one, [α]V +30.6° (c 0.17, CHCl 3 ).

c) By treating 0.099 g of the product from b) as described in example lj), 0.111 g (89%) of

[1(R*) , 1(3E) , (3a/8,7aa) ]-3,3a,5,6,7,7a-hexahydro-1-(1,5-dimethyl-5-[(trimethylsilyl) oxy]-3-hexenyl)-7α-methyl-4H-. inden-4-one, [α]V +26.4° (c 0.22, CHCl 3 ). d) As described in example lk), starting from 0.265 g of [3S-(1Z,3a,5/3)]-[2-[3,5-bis[[(1,1-dimethyl)dimethylsilyl]oxy ]-2-methylenecyclohexylidene]ethyl]diphenylphosphine oxide and 0.095 g of the product from c), 0.162 g (83%) of (lj3,3a,5Z,7E,23E)-1,3-bis[ [ (1,1 -dimethylethyl)dimethylsilyl]oxy]-25-[(trimethylsilyl)oxy]-9,10-secocholesta-5,7,10(19),16,23-pentaene, MS m/e 712 (M+) . e) By treating 0.159 g of the product from d) as in example 11), 0.077 g (84%) of 1,25-dihydroxy-16,23E-bisdehydrocholecalciferol is obtained, [a]V +46.5° (c 0.20, MeOH).

Example 4

a) As described in example lk), starting from 0.225 g of [5S-(1Z)]-[2-[5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-2-methylenecyclohexylidene]ethyl]diphenylphosphine oxide and 0.110 g of the product from example 3c), 0.150 g (81%) of (3/3,5Z,7E,23E)-3-[ [ (1,1-dimethylethyl)-dimethylsilyl]oxy]-25- [(trimethylsilyl)oxy]-9,10-secocholesta-5,7,10(19), 16, 23-pentaene, [ct]V <+>68.3° (c 0.18, CHCl 3 ). b) By treating 0.144 g of the product from a) as described in example 11), 0.076 g (78%) of 25-hydroxy-16,23E-bisdehydrocholecalciferol is obtained, [a]22 ~*"62.5° ( c 0.20, MeOH).

Example 5

a) To a solution of 6.25 g of ethyl 3-bromopropionate in 28 ml of THF at -20°C, add 28.8 ml of 2.8 M methylmagnesium bromide

in ether. The mixture is stirred at room temperature for 170 min. After adding 15 ml of saturated aqueous ammonium chloride and 42 ml of 1N HCl, the organic phase is separated and the aqueous phase is extracted with ether. The organic extracts are washed with saturated salt solution, dried and evaporated. The residue is chromatographed on silica gel with 30% ethyl acetate-hexane to give 2.57 g (45%) of 4-bromo-2-methyl-2-butanol, MS m/e 151 (M 2 -CH 2 ).

b) To a solution of 2.56 g of 4-bromo-2-methyl-2-butanol and 4.86 g of imidazole in 15 ml of N,N-dimethylformamide at 0°C is added

6.48 g of chlorotriethylsilane. The mixture is stirred at room temperature for 200 min. After adding ice, the mixture is diluted with water and extracted with pentane. The organic phase is washed with water and saturated salt solution, dried and evaporated. The residue is chromatographed on silica gel with pentane to give 4.02 g (93%) of (3-bromo-1,1-dimethyl-propoxy)triethylsilane, MS m/e 265 (M + -CH 3 ).

c) To a solution of 0.930 g of [ 1 (R*), 3aR* (3a/3, 4et, 7aa) ] - 3a, 4 , 5, 6, 7, 7a-hexahydro-4-hydroxy-/3, 7α-dimethyl-3H-indene-1-ethanol-4-methyl-benzenesulfonate and 1.10 g of imidazole in 73 ml of methylene chloride at 0°C are added to 0.580 g of chlorotriethylsilane. The mixture is stirred at room temperature for 1.5 hours. After adding ice, the mixture is diluted with water and stirred for 20 min. The organic layer is extracted with methylene chloride. The extracts are washed with water, IN N 2 SOA, saturated aqueous NaHCO 3 and saturated saline. After drying and evaporation, the residue is purified on silica gel with ethyl acetate-hexane (1:5) to give 1.22 g (100%) of [1 (R*), 3aR*-(3a/3,4a,7aa) ]- 3a,4,5,6,-7, 7a-hexahydro-4-[ (triethylsilyl)oxy] -/3, 7a-dimethyl-3H-indene-1-ethanol-4-methylbenzenesulfonate, [a]<2>D ° +4 67l ° (c 0.31, CHCl 3 ). d) 0.282 g of magnesium is added to a solution of 3.08 g of (3-bromo-1,1-dimethyl-propoxy)triethylsilane in 31 ml of THF. The mixture is heated at 68°C for 3.5 hours. Then a mixture of 0.686 g of copper(I) iodide and the above-mentioned Grignard solution is stirred at 3°C for 30 min. To this is added a solution of 1.02 g of the product from c), and the mixture is stirred at room temperature for 40 min. After adding a mixture of ice and water, the mixture is extracted with ether. The organic phase is washed with 1N H 2 SOA and saturated aqueous NaHCO 3 , dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:15) to give 1.80 g of [1(R*), 3aR*-(3aØ, 4a, 7aa)]-3a, 4, 5, 6, 7, 7α-hexahydro-1-[1,5-dimethyl-5-[(triethylsilyl)-oxy]hexyl]-4-[(triethylsilyl)oxy]-7α-methyl-3H-indene, MS m/e 479 (MVEt) . e) To a solution of 1.60 g of the product from d) in 5 ml of THF, 2.00 ml of 1M tetrabutylammonium fluoride in THF is added.

The mixture is heated at 68°C for 50 min. After cooling to room temperature, the mixture is diluted with water and extracted with methylene chloride. The organic phase is washed with salt solution, dried and evaporated. The residue is purified on silica gel with ethyl acetate-hexane (1:1) to give 0.420 g (79%) of [1(R*), 3aR*-(3aØ,4aa,7aa)]-3a,4,5,6, 7,7α-Hexahydro-4-hydroxy-α,α-ε,7α-tetramethyl-1H-indene-1-pentanol, [α]V = +12.0° (c 0.25, CHCl 3 ).

f) To a solution of 0.210 g of the product from e) in 18 ml of methylene chloride, add 0.870 g of pyridinium dichromate and 44 mg

pyridinium p-toluenesulfonate. The mixture is stirred for 175 min. After adding 50 ml of ether, the mixture is stirred for 5 min. and filtered. The solids are washed with saturated aqueous CuSOA, water, half-saturated aqueous NaHCO 3 and saturated saline.

The organic phase is dried and evaporated. The residue is chromatographed on silica gel with 35% ethyl acetate-hexane to give 0.175 g (84%) of [ 1 (R*), 3aR*-(3a/3, 7aa) ]-3 , 3a~5, 6, 7, 7a -hexahydro-1-(5-hydroxy-1,5-dimethylhexyl)-7a-methyl-4H-inden-4-one, [a] V +28.2° (c 0.22, CHCl 3 ).

g) By treating 0.168 g of the product from f) as described in example lj), 0.211 g (100%) of

[ 1 (R*) , 3aR*-(3a/3, 7aa) ] -3, 3a, 5, 6,7, 7a-hexahydro-1-(1,5-dimethyl-5-[(trimethylsilyl)oxy] hexyl)-7α-methyl-4H-inden-4-one, [α]<2>D° <+>21.9° (c 0.27, CHCl 3 ). h) As described in example lk), starting from 0.581 g of [3S-(1Z,3a,5/3)]-[2-[3,5-bis[[(1,1-dimethyl)dimethylsilyl]oxy ]-2-methylenecyclohexylidene]ethyl]diphenylphosphine oxide and 0.210 g of the product from g), 0.358 g (83%) of (1a,3j3,5Z,7E)-1,3-bis [ [ (1,1-dimethylethyl ) dimethylsilyl ] oxy ] - 25-[(trimethylsilyl)oxy]-9,10-secocholesta-5,7,10(19),16-tetraene, MS m/e 714 (M+) .

i) Treatment of 0.350 g of the product from h) as described in example 11), 0.168 g (83%) of 1,25-dihydroxy-16-dehydrocholecalciferol is obtained, [a]<2>D° +40.0° (c 0.17, MeOH).

Example 6

a) As described in example lk), starting from 0.383 g of [5S-(1Z)]-[2-[5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-2-methylenecyclohexylidene]ethyl]diphenylphosphine oxide and 0.188 g of the product from example 5g), 0.245 g (78%) of (3a,5Z,7E)-3-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-25-[(tri-methylsilyl ) oxy]-9,10-secocholesta-5,7,10(19),16-tetraene, [α]V +67.5° (c 0.20, CHCl3) . b) Treatment of 0.239 g of the product from a) as described in example 11) gives 0.135 g (83%) of 25-hydroxy-16-dehydro-cholecalciferol, [a]2D3 +75.4° (c = 0.13 , MeOH).

The following examples A and B illustrate the composition of soft gelatin capsules for oral administration and of a cream for topical application:

Claims (2)

1. Analogy method for making "of a therapeutically active compound with the formula wherein R is hydrogen or hydroxy and A is -C=C-, -CH=CH- with E-configuration or -CH2-CH2-, characterized in that the method comprises reacting a corresponding compound of formula I which instead of the two or three the hydroxy groups contain two or three protected hydroxy groups with the formula wherein R x and R 3 are C 1 -alkyl and R 2 is C 1 -alkyl, aryl or aryl-C 1 -C 1 -alkyl, with an agent capable of removing the protecting groups. 2. Process according to claim 1 for the production of: 1,2 5-dihydroxy-16-dehydro-2 3-didehydrocholecalciferol, 1,25-dihydroxy-16,23E-bisdehydrocholecalciferol or 1,25-dihydroxy-16-dehydrocholecalciferol, characterized by that correspondingly substituted starting materials are used.