NL8520265A - DERIVATIVES OF VITAMIN D AND METHODS FOR PREPARING THE SAME - Google Patents

Description

- 1 - 8 52 0 2 65

Vitamin D derivatives and methods for their preparation.

This invention concerns new vitamin D derivatives. More particularly, this invention relates to 24-homo-vitamins. Even more particularly, this invention relates to hydroxylated 24-homovitamines.

Vitamin D is known to regulate calcium and phosphorus metabolism in humans and animals, and it has now been established that the biological activity of vitamin D is dependent on its metabolic conversion in vivo to hydroxylated derivatives. For example, vitamin D1 is in vivo hydrolysed in the liver to 25-hydroxy-vitamin D1, which in turn is converted into 1,25-dihydroxyvitamin D1 in the kidneys. It is the latter compound that is now recognized as the circulating hormonal form of Vitamin D.

Because of their biological influence on promoting calcium and phosphorus transport in the gut and mobilizing and mineralizing the bones, these forms of vitamin D are important pharmaceutical products which are excellent for the treatment of various bone diseases. Background Art Vitamin D derivatives and their preparation and application have been discussed in many places in patents and other literature. For example, U.S. Patent No. 3,565,924 is directed to 25-dihydroxycholecalciferol, U.S. Patent No. 3,697,559 to 1,25-dihydroxy-25 cholecalciferol, U.S. Patent No. 3,741,996 to Ια-hydroxycholecalciferol, U.S. Patent 3,786,062 to 22-dehydro-25-hydroxycholecalciferol, U.S. Patent 3,880,894 to 1,25-dihydroxyergocalciferol, U.S. Patent 4,201,881 to 24.24-difluoro-1a, 25-30 dihydroxycholecalciferol, and U.S. Patent 8520265 - 2 - 4,196,133 to 24,24-difluoro-1a, 25-dihydroxycholecalciferol. Definition of the invention

New derivatives of vitamin have now been found which exhibit an excellent vitamin D-like effect 5 and which for this reason can very well serve as a replacement for vitamin as well as the various derivatives thereof in known applications, such as the treatment of various disorders in which a calcium and phosphorus disturbance, such as hypoparathroidism, osteodystrophy, osteomalacia and osteoporosis.

These derivatives are 24-homovitamines, and more particularly 1a, 25-dihydroxy-22E (o 2) -dehydro-24-homo-vitamin D 1 and 1, 25-dihydroxy-24-homovitamine D 1.

The compounds of this invention may be suitably represented by Formula 1, wherein R 1 and R 15 are each selected from the group of hydrogen, acyl groups of 1 to 4 carbon atoms and benzoyl, and wherein and R 4 each represent a hydrogen atom or together a second carbon-carbon bond.

Best embodiment of the invention.

The compounds of the invention can be prepared according to a method shown in the accompanying reaction scheme and the description which now follows. In the schematic and detailed description of the method, like numbers indicate like compounds.

In the process of this invention, bisnorcholenic acetate (a) was reduced with lithium aluminum hydride and then oxidized with dichlorodicyan benzoquinone to give 1,4,6-trienone-3 (b) in 47%.

The 22-THP ether of (b) was treated with alkaline hydrogen peroxide to give the 1,2a epoxide (1) in 41%. Reduction of (1) with lithium and ammonium chloride at -78 ° in liquid ammonia / tetrahydrofuran and then treatment with chloromethyl methyl ether gave the dimethoxymethyl ether (2) in 38% yield. Removal of the THP group followed by a Swern oxidation gave the aldehyde (4) in 81% yield. This was reacted with vinyl magnesium bromide to obtain 8520265-3 allyl alcohol (5) in 94% yield. This alcohol was reacted in boiling xylene with a catalytic amount of propionic acid with triethyl orthoacetate to give the ester (6) in 93% yield. The ester (6) was then reacted with methyl magnesium bromide to give the alcohol (7) in 93% yield. Removal of the MOM group followed by acetylation gave the (22E) -la, 3f3-diacetoxy-25-hydroxy-24-homocholesta-5,22-diene (9) in 73% yield.

Allyl bromination of (9) with N-bromosuccinimide, followed by treatment with tetra-n-butylammonium bromide and then with tetra-n-butylammonium fluoride gave the 5,7,22-triene (10) as the main product in 24% yield. The 5,7-diene (10) was irradiated in benzene-ethanol with a medium-pressure mercury lamp for 5 minutes and then refluxed for 1 hour, and finally hydrolysed to give the (22E) -la, 25-dihydroxy- in 22% yield. 22-dehydro-24-homovitamine (11).

The 5,22-diene (9) was selectively hydrogenated to 5-one (10) in 92% yield. This compound was converted to 1,25-dihydroxy-24-homovitamin D3 (14) in a total yield of 12% via the 5,7-diene (13) in the manner already described.

Detailed description of the method

In the following detailed description of the method of the invention, the melting points were determined under a heated center microscope; they have not been corrected. The H-NMR spectra were recorded with a Hitachi R-24A (60 MHz) in CDCl 2 with Me 2 Si as the internal standard unless otherwise indicated. Mass spectra were obtained with a Shimadzu QP-1000 mass spectrometer at 70 eV. UV spectra were taken in ethanol solution in a double beam spectrophotometer Shimadzu UV-200. Column chromatography was done on silica gel (E. Merck, Kieselgel 60, 62-2104 un). Thin layer preparative chromatography was performed on pre-coated silica gel plates (E. Merck, Kieselgel 60, F254, 0/25 mm thick). The "usual work-up" involves dilution with water, extraction with an organic solvent stated in brackets, washing the 8520265-4 extract to neutral, drying over anhydrous magnesium sulfate, filtering and removing the solvent under reduced pressure. The following abbreviations apply: THP - tetrahydropyranyl, THF - tetrahydrofuran, ether - diethyl ether, MeOH - methanol, 5 MOM - methoxymethyl. The temperatures are in ° C.

22-hydroxy ~ 23,24-dinorchola-1,4,6-trienone-3 (b)

To a solution of 7.0 g (18.04 mg mol) of 38-acetoxydinorcholenic acid (a) in 20 ml of THF was added 3.0 g (78.95 mg mol) of lithium aluminum hydride. The mixture was stirred at 60 ° C for 14 hours. Water and ethyl acetate were carefully added to this reaction mixture. Filtration and removal of the solvent gave 5.2 g of residue. This was treated in 140 ml of dio-xane with 11.7 g (51.54 mg mol) of dichlorodicyanahbenzoquinone, which was refluxed for 14 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate evaporated to dryness to give a residue which was applied to a column of 200 g of aluminum oxide. Eluting with dichloromethane gave 2.8 g (47%) of trienone (b) with mp. 156-157 ° (from ether), UV in EtOH λ y in nm (ε) at 299 (13000), 252 20 (9200), 224 (12000), 1 H NMR (in CDCl-j) δ = 0.80 (3H, s, 18-H3), 1.04 (3H, d, J = 6 Hz, 21-H3), 1.21 (3H, s, 19-H3), 3.10-3.80 (3h, m , 22-H2 and OH), 5.90-6.40 (4H, m, 2-H, 4-4H, 6-H and 7-H), 7.05 (1H, d, J = 19 Hz, 1- H), MS m / z: 326 (M +), 311, 308, 293, 267, 112.

25 la, 2a-epoxy-22-tetrahydropyranyloxy-23,24-dinorchola-4,6-dienon-3 (1)

2.7 g (8.28 mg mol) in 50 ml dichloromethane of the alcohol (b) was treated for 1 hour at room temperature with 1.5 ml (16.42 mg mol) dihydropyran and 50 mg p-toluenesulfonic acid. The usual work-up (with ethyl acetate) gave a crude product. To a solution of this product in 70 ml of MeOH, 4.8 ml of 30% and m - 10% NaOH / MeOH were added, and this mixture was stirred at room temperature for 14 hours. The usual work-up (with ethyl acetate) gave a crude product which was applied to a column of 50 g of silica gel. Eluting with benzene / ethyl acetate 100: 1 gave 1.45 g (41%) of epoxide (1) with 852 g of 2% 5-5 m.p. 113-115 ° (from hexane) UV in EtOH λ in nm (ε) at 290 (22000), h-NMR (in CDCl3) δ = 0.80 (3H, s, 18-H3), 1.07 (3H , d, J = 6 Hz, 21-H3), 1.18 (3H, s, 19-H3), 3.38 (1H, dd, J = 4 and 1.5 Hz, 1-H), 3, 55 (1H, d, J = 4 Hz, 2-H), 3.30-4.10 (4H, m, 5 22 “H2 and THP) '4.50 (1H, m / THP), 5.58 (1H, d, J = 1.5 Hz, 4-H), 6.02 (2H, s, 6-H and 7-H), MS m / z: 342 (M + - DHP), 324 (M + -THPOH) , 309, 283, 85.

1a, 3β-dimethoxymethoxy-23,24-dinorchol-5-en-22-tetrahydropyranyl ether (2). 10 In 200 ml of liquid ammonia at -78 ° were placed under argon 5.00 g of lithium over 30 minutes. After stirring at -78 ° for 1 hour, 2.00 g (4.69 mgmol) of la, 2a-epoxy-22-tetrahydropyranyloxy-23,24-dinorchola-4,6-dienon-3 (1) was added over 30 minutes 150 ml of dry THF was added dropwise and this mixture was stirred at -78 ° for 1 hour. 60 g of anhydrous NH 4 Cl were added in small bits over 1 hour to this reaction mixture. After 1.5 hours, the cooling bath was removed and most of the ammonia was removed by bubbling argon through it. The usual work-up (with ether) gave a crude product. This was treated with 2.0 ml (26.34 mg mol) chloromethyl methyl ether and 4.6 ml (24.93 mg mol) Ν, Ν-diethylcyclohexylamine in 20 ml dioxane for 24 hours at 45 ° C. The usual work-up (ethyl acetate) gave a crude product which was applied to a column of 40 g of silica gel.

Eluting with hexane / ethyl acetate 5: 1 gave 922 mg (38%) dimethoxymethyl ether (2) as an oil. H-NMR: δ = 0.70 (3H, s, 18-H3), 1.02 (3H, s, 19-H3), 1.04 (3H, d, J = 6 Hz, 21-H3), 3, 34 (3H, s, -0-CH3), 3.37 (3H, s, -0-CH3), 4.63 (2H, ABq, J = 7 Hz, ΔΑΒ = 11 Hz, 1a-0-CH2- 0-), 4.64 (2H, s, 3B-O-CH 2 -O-) and 5.50 (1H, m, 6-H).

30 la, 3g-dimethoxymethoxy-23,24-ά: ΙηοΓθίιο1-5-θηο1-22 (3)

Of the THP ether (2), 922 mg (1.77 mg mol) in 8 ml THP and 8 ml MeOH was treated with 1 ml 2M HCl at room temperature for 2 hours. The usual work-up (ethyl acetate) gave a crude product which was applied to a column of 40 g of silica gel. Eluting with hexane / ethyl acetate 2: 1 gave 678 mg (88%) of the alcohol (3) as an amorphous substance. ^ H-NMR: δ = 0.70 (3H, s, 8520265 - 6-18 "H3), 1.02 (3H, s, 19-H3), 1.04 (3H, d, J = 6 Hz, 21-H3), 3.34 (3H, s, -0-CH3), 3.38 (9H, s, -0-CH3), 4.65 (2H, ABq, J = 7 Hz, ΔΑΒ = 11 Hz , 1a-O-CH 2 -O-), 4.66 (2H, s, 3B-O-CH 2 -O-), 5.53 (1H, m, 6-H).

5 la, 3g-dimethoxymsthoxy-23,24-dinorchol-5-enal-22 (4)

To a solution of 0.27 ml (3.09 mg mol) oxalyl chloride in 8 ml dichloromethane was added at -78 ° under argon 0.44 ml (6.21 mg mol) dimethyl sulfoxide. The mixture was stirred at -78 ° C for 10 minutes. To this solution was added 660 mg (1.51 mgmol) of alcohol (3) in 5 ml of dichloromethane at 10 -78 °. After stirring for 15 minutes, 1.89 ml (13.6 mg mol) of triethylamine was added. The mixture was stirred at -78 ° C under argon for 5 minutes and then allowed to warm to room temperature. The usual work-up (with ether) gave a crude product which was applied to a column of 30 g of silica gel.

Eluting with hexane / ethyl acetate 4: 1 gave 607 mg (92%) aldehyde (4) as crystals with m.p. 71-72 ° (from hexane), 1 H-NMR, δ = 0.74 (3H, s, 18-H3), 1.04 (3H, s, 19-H3), 1.12 (3H, d, J = 6 Hz, 21-H3), 3.35 (3H, s, -0-CH3), 3.39 (3H, s, -0-CH3), 20 3.7 (1H, m, Ιβ-H ), 4.65 (2H, ABq, J = 7 Hz, ΔΑΒ = 11 Hz, 1a-0-CH2-0-), 4.66 (2H, s, 38-0-CH-0-), 5, 52 (1H, m, 6-H) and 9.61 (1H, d, J = 3Hz, -CHO), Anal. Ber. for ^ 26Ξ42 ° 5: C 71.85%, H 9.74%, Found: C 71.71%, H 9.68%. 1a, 3g-dimethoxymethoxychola-5,23-dienol-22 (5). To 70 mg (2.92 mg atom) magnesium in 3 ml THF was added 0.42 ml 50% vinyl bromide in THF (2.98 mg mol). The mixture was stirred at room temperature under argon for 30 minutes. 595 mg (1.37 mg mol) aldehyde (4) in 3 ml THF was added to the resulting Grignard reagent at room temperature. The mixture was stirred at room temperature for 1 hour. Usual work-up (with ether) gave a crude product which was applied to a column of 30 g of silica gel. Eluting with hexane / ethyl acetate 3: 1 gave 595 mg (94%) of the allyl alcohol (5) as an amorphous substance. 1 H-NMR δ = 0.70 (3H, s, 18-H3), 1.02 (3H, s, 19-H3), 35 3.35 (3H, s, -0-CH3), 3.38 ( 3H, s, -0-CH3), 3.69 (1H, m, Ιβ-H), 4.20 (1H, m, 22-H), 4.64 (2H, ABq, J = 7 Hz, ΔΑΒ = 11 Hz, 8520265 - 7 - 1a-O-CH ^ -O-), 4.65 (2H, s, 3 $ -0-CH2-0-), 5.52 (1H, m, 6-H) , 4.90-6.0 (3H, m, 23-H and 24-H2).

(22E) -la, 38-dimethoxymethoxy-27-norcholesta-5,22-dien-26-carboxylic acid ethyl ester (6) 5 A solution of 590 mg (1.28 mgmol) allyl alcohol (5), 1.0 ml (5.46 mgmol) triethyl orthoacetate and 4 drops of propionic acid in 8 ml xylene were refluxed for 2 hours under argon. Removal of the solvent under reduced pressure gave a residue which was applied to a column of 10 g of silica gel. Eluting with hexane / ethyl acetate 4: 1 gave 630 mg (93%) of ester (6) as an oil. ^ H-NMR: δ = 0.68 (3H, s, 18-H3), 0.97 (3H, d, J = 6 Hz, 21-H3), 1.03 (3H, s, 19-H3) , 1.24 (3H, t, J = 7 Hz, -CC> 2CH2CH3), 3.35 (3H, s, -0-CH3), 3.39 (3H, s, -0-CH3), 3, 70 (1H, m, Ιβ-H), 4.11 (2H, 15 q, J = 7 Hz, -C02CH2CH3), 4.64 (2H, ABq, J = 7 Hz, ΔΑΒ = 11 Hz, la-0 -CH2-0-), 4.65 (2H, s, 38-0-CH2 ~ 0-), 5.29 (2H, m, 22-H and 23-H), 5.52 (1H, m, 6-H).

If desired, the 22E stereoisomer, compound (6) can be easily converted to the 22Z stereoisomer by iodine treatment. For example, treatment under diffuse daylight of compound (6) in ether with a catalytic amount (2% based on (6)) iodine gave an isomerization of trans to cis, so that after purification by HPLC (a column of Zorbax-Sil of 4 6 x 25 cm, with 6% isopropanol in hexane) had the 22Z stereoisomer.

(22E) -la, 3B-dimethoxymethoxy-24-homocholesta-5,22-dienol-25 (7)

To a solution of 605 mg (1.14 mg mol) ester (6) in 6 ml THF was added 4.5 ml 1 M (4.5 mg mol) methyl magnesium bromide in THF at room temperature. The mixture was stirred at room temperature for 1 hour. Usual work-up (with ether) gave a crude product which was applied to a column of 30 g of silica gel. Eluting with hexane / ethyl acetate 3: 1 gave 548 mg (93%) of alcohol (7) as an oil. H-NMR: δ = 0.68 (3H, s, 18-H3), 0.97 (3H, d, J = 6 Hz, 21-Hj), 1.01 (3H, s, 35 1/21 ( 6H, s, 26-H3 and 27-Hj), 3.33 (3H, s, -0-CH3), 3.38 (3H, s, —0-CH3), 3.70 (1H, m, 18 -H), 4.64 (2H, ABq, J = 7 8520265 - 8 -

Hz, MB = 11 Hz, 1a-0-CH2-0-), 4.65 (2H, s, 3 $ -0-CH2 ~ 0-), 5.29 (2H, m, 22-H and 23- H) and 5.50 (1H, m, 6-H).

(22E) -24-Homocholesta-5,22-dien-la, 3β, 25-triol (8)

A solution of 540 mg (1.04 mg mol) of di-5 methoxymethyl ether (7) in 15 ml of THF was treated with 3 ml of 6M HCl at 50 ° C for 2 hours. Usual work-up (with ethyl acetate) gave a crude product which was applied to a 20 g column of silica gel. Eluting with hexane / ethyl acetate 1: 1 gave 428 mg (95%) of the triol (8) as crystals with m.p. 164-166 ° C (from hexane / 10 ethyl acetate), 1 H-NMR δ = 0.68 (3H, s, 18-H3), 0.95 (3H, s, J = 6 Hz, 21-H3), 1.00 (3H, s, 19-H3>, 1.20 (6H, s, 26-H3 and 27-H3), 3.80 (1H, m, Ιβ-H), 3.92 (1H, m .3a-H), 5.30 (2H, m, 22-H and 23-H) and 5.53 (1H, m, 6-H).

(22e) -la, 3g-diacetoxy-25-hydroxy-24-homocholesta-5,22-diene (9) A solution of 395 mg (0.919 mgmol) triol (8) in 2 ml pyridine was left at room temperature with 1 ml of acetic anhydride treated. Usual work-up (with ethyl acetate) gave a crude product which was applied to a column of 20 g of silica gel. Eluting with hexane / ethyl acetate 2: 1 gave 361 mg (77%) diacetate (9) as an oil. * H-NMR: δ = 0.67 (3H, s, 18-H3), 0.97 (3H, d, J = 6 Hz, 21-Hj), 1.07 (3H, s, 19-H3) , 1.21 (6H, s, 26-H3 and 27-H3), 201 (3H, s, acetyl), 2.04 (3H, s, acetyl), 4.98 (1H, m, 3a-H) , 5.05 (1H, m, Ιβ-H), 5.31 (2h, m, 22-H and 23-H) and 5.52 (1H, m, 6-H).

25 (22E) -1a, 3 g-diacetoxy-25-hydroxy-24-homocholesta-5,7,22-triene (10)

A solution of 51 mg (0.0992 mg mol) of the 5-one compound (9) and 21 mg (0.118 mg mol) of N-bromo succinimide in 3 ml of carbon tetrachloride was refluxed for 20 minutes under argon. After the mixture had cooled to 0 ° C, the resulting precipitate was filtered off. The filtrate was concentrated to a residue below 40 ° C. This was treated with a catalytic amount of tetra-n-butylammonium bromide in 5 ml THF for 50 minutes at room temperature.

Then the mixture was treated with 3.5 ml of 35 (3.5 mg mol) tetra-n-butylammonium fluoride in THF for 30 minutes at room temperature.

Conventional work-up (with ethyl acetate) gave a crude product which was subjected to preparative thin layer chromatography (8520265-9) (developed with hexane / ethyl acetate 4: 1, 5x). The Rf 0.48 band was scraped off and extracted with ethyl acetate. Removal of the solvent gave 12.5 mg (24%) of the 5,7-diene (10), µV in EtOH λ at 293, 282 and 271 nm.

max J

1a, 25-dihydroxy-22E-dehydro-24-homovitamine D ^ (11)

A solution of 7.3 mg (0.0143 mgmol) of 5,7-diene (10) in 90 ml of benzene and 40 ml of ethanol was irradiated with a medium pressure mercury lamp at 0 ° C under argon at 0 ° C for 5 minutes. The reaction mixture was refluxed for 1 hour under argon. Removal of the solvent under reduced pressure gave a crude product, which was subjected to preparative thin layer chromatography (developed with hexane / ethyl acetate 4: 1.5 times). The band with an Rf value of 0.38 was scraped off and extracted with ethyl acetate. Removal of the solvent gave 1.8 mg (25% of the vitamin D 2 diacetate. The Rf 0.43 band was also scraped off and extracted with ethyl acetate. Removal of the solvent gave 2.1 mg (29%) of the 5,7-diene (10).

The 1.8 mg (2.15 µmol) vitamin D ^ diacetate was treated in 4 ml THF with 1 ml 5% KOH / MeOH at room temperature for 20 minutes. Usual work-up (with ethyl acetate) gave a crude product which was subjected to preparative thin layer chromatography (hexane / ethyl acetate 1: 2, developed 3x). The band with Rf value 0.43 was scraped off and extracted with ethyl acetate. Removal of the solvent gave 1.4 mg (90%) of vitamin D 1 analog (11). For the purity of the product (11), 100% was found by high performance liquid chromatography (a Shimadzu column LC-3A, Zorbax Zil as normal 30 phase, 4.6 mm inner diameter, 15 cm length; running liquid

MeOH / CH2Cl2 1:49, flow rate 3 ml / min, retention time 11.5 min). The vitamin D 1 analog (11) had the following spectrum: UV in EtOH λ at 265 nm, λ. at 228 nm, MS m / z: 428 (M +), max J min 410, 392 (base peak), 374, 287, 269, 251, 152, 134, 123, 59, 35 H NMR (360 MHz) 6 = 0.55 (3H, s, 18-Hj), 1.02 (3H, d, J = 6.6 Hz, 21-H3), 1.22 (6H, s, 26-H3 and 27-Hj) , 2.32 (1H, dd, J = 8520265 - 10 - 13.2 and 6.7 Hz), 2.60 (1H, dd, J = 13.0 and 3.0 Hz), 2.83 (1H , dd, J = 12.0 and 3.0 Hz), 4.23 (1H, m, W ^ 2) = 18.4 Hz, 3ct-H), 4.43 (1H, m, = 16.9 Hz, Ιβ-H), 5.00 (1H, bs, W ^ 2 = 3.2 Hz, 19-H), 5.30 (1H, dd, J = 15.0 and 7.1 Hz, 22- H or 23-H), 5.33 (1H, bs, W ^ 2 = 3.2 Hz, 19-H), 5.37 (1H, dd, J = 15.0 and 5.8 Hz, 22 -H or 23-H), 6.01 (1H, d, J = 11.0 Hz, 7-H), 6.32 (1H, d, J = 11.0 Hz, 6-H).

1a, 3g-diacetoxy-24-homocholest-5-enol-25 (12)

A mixture of 40 mg of 5,22-diene (9) and 10 mg of 10% Pd-C in 2 ml of ethyl acetate was stirred under hydrogen at room temperature for 3 hours. The Pd catalyst was filtered off and the filtrate concentrated to leave a residue which was applied to a column of 5 g of silica gel.

Eluting with hexane / ethyl acetate 4: 1 gave 37 mg (92%) of the 5-ene compound (12) as oil. H-NMR δ = 0.66 (3H, s, 18-Hg), 1.08 (3H, s, 19-H3), 120 (6H, s, 26-H3, and 27-H3), 2.02 ( 3H, s, acetyl), 2.05 (3H, s, acetyl), 4.97 (1H, m, 3a-H), 5.07 (1H, m, 18-H), 5.53 (1H, m, 6-H).

1a, 3g-diacetoxy-24-homocholesta-5,7-dienol-25 (13) Of the 5-one compound (12), 19 mg (0.037 mgmol) was prepared in the manner described in (5) above, 7-diene (13) converted (5.8 mg, 31%).

UV in EtOH: λ at 293, 282 and 271 nm. max 1a, 25-dihydroxy-24-homovitamine (14) The 5.8 mg (0.0113 mg mol) of 5,7-diene (13) was added in the vitamin D3-analog (14) in the manner described for (11). converted (890 µg, 19%). The retention time of (14) in the HPLC described above was 11.0 min. UV in EtOH λ at 265 max nm, λ. at 228 nm. MS m / z 430 (M +), 4.12, 394 (base peak), min 30 376, 287, 269, 251, 152, 134, 59.

If desired, the compounds of this invention can be easily obtained in crystalline form by crystallization from suitable solvents, eg hexane, ethers, alcohols or mixtures thereof, as will be apparent to those skilled in the art.

852 0 2 6 5 - 11 -

Biological action

Mobilizing bone calcium by 1,25- (OH) ^ - 24-homo-O ^

The mobilizing effect on bone calcium was determined by measuring the increase in serum calcium level in response to the administered compound. Male weaned rats (Holtzman Co. of Madison, WI) were fed a low-calcium, vitamin D-deficient diet (Suda et al. In J. Nutr. 100 (1970) 1049-1050) and water ad libitum for 3 weeks. divided into three groups of 5-6 rats each and received either intrajugular or 1.25- (OH) or the test substance dissolved in 0.05 ml of 95% ethanol. Rats of the blank group received 0.05 in the same manner ml ethanol 18 hours after the dose, the rats were sacrificed and their blood was collected and centrifuged to give the serum Calcium concentrations in the serum were determined with an atomic absorption spectrometer Model 403 (Perkin-Elmer Co. of Norwalk , Conn.) In the presence of 0.1% lanthanum chloride.

The results are presented in the following table: Table 1 20 Increase in the serum concentration of calcium in response to the administered substance.

Administered Administered how- Serum-calcium compound quantity concentration _ (pmol / rat) _ (mg / 100 ml)

Test I Ethanol - 3.6 ± 0.3 5 *! 25 la, 25- (OH) 2D 650 4.9 ± 0.2 7 'la, 25- (OH) 2-24 "650 4.4 ± 0.2 homo-D ^

Test II Ethanol - 4.2 ± 0.1 la, 25- (OH) 2D_ 325 5.0 ± 0.5 la, 25- (OH) -22E- 650 5.0 ± 0.5} 2 q dehydro- 2C “homo-D ^ * Standard deviation difference from b) to a) p <0.001 and d) from c)

From the foregoing data, it can be concluded that in the vitamin D-responsive system of vitamin D deficient animals, the compounds of this invention exhibited the same activity as the 1,25-dihydroxy vitamin, the circulating hormonal form of the vitamin, although in the case of the 22-dehydro derivative the dosage was clearly higher.

The compounds of this invention can be readily administered as sterile solutions by injection or intravenously, or else in the form of oral nutritional preparations, or otherwise by suppositories, and even transcutaneously. Doses of from about 0.1 µg to about 2.5 µg per day are effective in achieving a physiological response in the calcium balance characteristic of the vitamin D-like activity, maintaining a dose of about 0.1 µg to about 0.5 µg is suitable.

Dosage forms of the compounds can be prepared by combining them with a non-toxic, pharmaceutically acceptable carrier, as is well known in the art. Such carriers can be solid or liquid, for example, corn starch, lactose, sucrose, peanut oil, olive oil, sesame oil and water. When a solid carrier is used, the dosage forms of the compounds of the invention may be tablets, capsules, powders, troches and lozenges. When a liquid carrier is used, soft gelatin capsules, syrups and liquid suspensions, emulsions or solutions can be the dosage forms. The dosage forms may also contain adjuvants such as preservatives, stabilizers, wetting agents and emulsifiers, solubilizers, etc.

They can also contain other therapeutically valuable substances.

It is to be understood that although dosages herein are mentioned, the doses to be administered to a patient in any given case will depend on the condition to be treated, the desired end result, the size of the patient, and other factors known among those skilled in the art of therapeutic use of such drugs.

8520265

Claims (11)

1. Compounds of formula 1, wherein R 1, and R 1 are each selected from the group of hydrogen, 5 acyl groups of 1 to about 4 carbon atoms and benzoyl, and wherein R 1 and R 2 are both hydrogen atoms, or together a second carbon-carbon bond. A compound according to claim 1, wherein R 1, R 1 and R 8 are hydrogen and R 7 and R 1. are hydrogen atoms. The compound of claim 2 in crystalline form. The compound of claim 2 together with a pharmaceutically acceptable excipient. A compound according to claim 1, wherein R 1, R 1 and R 1 are hydrogen and R 1 and together form a second carbon-carbon bond. The compound of claim 5 in crystalline form. A compound according to claim 4 together with a pharmaceutically acceptable excipient. A compound according to claim 4 wherein the A22 bond has the E configuration. A compound according to claim 4 wherein the A22 bond has the Z configuration. 10. Compounds of formula 2 wherein R is selected from the group of hydrogen, acyl groups of 1 to 4 carbon atoms, benzoyl and methoxymethyl. 11. Compounds of formula 3, wherein R is selected from the group of acyl groups having 1 to 4 carbon atoms and benzoyl and wherein R 1 and R 1. represent hydrogen atoms or together form a second carbon-carbon bond. L - L "^".