DE2920092A1 - ANTIVITAMIN D COMPOUNDS AND PHARMACEUTICAL COMPOSITIONS THEREOF - Google Patents

Description

2S2ÖÖS2

WISCONSIN ALUMNI
RESEARCH FOUNDATION
614 North Walnut Street
Madison, Wisconsin 53707
V. St. A *

Antivitamin D compounds and pharmaceutical compositions containing them

Addendum to patent

(Patent application P 28 12 741.5)

The invention relates to compounds having antivitamin D activity and, more particularly, to analogues of vitamin D which, at the same time Administration of vitamin D to an animal can inhibit or reverse the normal response of the animal to vitamin D.

The use of various vitamins D to correct certain calcium metabolic dysfunctions, e.g. B. of rickets, has long been known. Recently, one enable various derivatives of vitamin D, such as 25-hydroxycholecalciferol (see US Pat. No. 3,565,924), oc-hydroxy-cholecalciferol (see US Pat. No. 3,741,996) and 1 ^ S-dihydroxy-cholecalciferol (cf. U.S. Patent 3,697,559) the treatment of other metabolic disorders affecting calcium and phosphorus balance (imbalance) in animals (see, for example, U.S. Patent Nos. 3,646,204, 3,649,596, and 3,879,548).

According to the invention, other vitamin D derivatives have now been found, that have an antivitamin D effect. In other words, such compounds tend to be administered concomitantly

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# ■

with vitamin D to reverse the normal response of an animal on vitamin D, d. H. they develop an inhibitory effect against intestinal calcium transport and mobilization of bone calcium, effects of vitamin D. This effect recommends its use to correct certain calcium disorders or »Calcium diseases in animals, such as hypercalcemia, hypervitaminosis D, hypersensitivity to vitamin D and metastatic calcification.

In the main patent (patent application P 28 12 741.5)

became a class of compounds of general formula

CH ₃

CfL

CH-CH., - CH., -X \ ■ λ 2

' CH,

described, wherein X is selected from the group of

CH

CH.

3
- R.

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-CH ₂ - O - R ₁

-CH ₂ - Si - R ₁

-CH ₂ -

■ ^R 2

- N

or

^R 3

wherein R ^ is each independently of one another lower-alkyl, V _r) and R- are each independently hydrogen, lower-7-.cyl, lower-alkyl or aryl and R is hydrogen, acyl with 1-6 carbon atoms or benzoyl. In particular vi ι d for the compound in which R is hydrogen, X has the meaning of -CH ₉ -N 2, wherein R ₉ and R-. each flethy] ]: e-

^ R ₃

interpret, referred to as 25-aza-vitamin D ^, a gui: e anti- ^T "it.ainiii D-effect described, and the compound is therefore suitable for use in reducing the reversal of the biological effect of vitamins D and in reducing it from Hypercal cainic =.

In the context of the present invention, other L-j.-f -.- targets were now le connections found for this purpose: v / er t ve J 1 Bind «

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These compounds have the general formula where R Is hydrogen and i) X has the meaning of

I ³

-CH ₂ -

wherein R _{1 is} methyl, referred to as 25-methyl-vitamin D ₃ , or ii) X the meaning of

8 / ^R 2

-C -

where R ₂ and R- are both methyl, referred to as cholanocalciferol-dimethylamide.

25-methyl-vitamin D ₃ can be produced according to the following reaction scheme:

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OTs

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.. * 7 -

AcO

13th

The preparation of the 25-methyl analogue 14 can therefore be carried out by a procedure which is similar to that used for the synthesis of 25-aza-vitamin D ₃ . The bromide 2 can thus be prepared as described in the main patent. The starting material can thus be prepared from stigma sterol by the procedure of Partrick et al (HeIv. Chim. Zicta 57, 764, 1974), and the bromide obtained therefrom by reaction with pulverized lithium bromide in acetonitrile. The reaction of the bromide 2 in THF with the lithium salt of 3,3-dimethylbutyne (made with n-butyl-lithium) gives the acetylenic intermediate 10. By hydrogenating 10 (H ₃ , 10% Pd / C), 11 is obtained, which is converted into 25-methylcholesterol-3ß-acetate (compound 12) by solvolysis in hot glacial acetic acid. This intermediate acid is allylic brominated and sum 5,7-diene 13 dehydro-

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brominated. By irradiating 13 (3ß-acetoxy-25-methyl-cholesterol-5,7-diene) followed by thermal isomerization of the isolated previtamin derivative and saponification (KOH / MeOH) you get the 25-methyl-vitamin D_ (14).

6β-methoxy-25-methyl-3 u. , 5-cyclo-5 o6-cholest-23-yne (10)

25 ml of a 1.5 M solution of n-butyllithium in hexane become a solution of 3 g of 3,3-dimethyl-1-butyne in 120 ml of dioxane 5 ° joined. The mixture is stirred for 4 hours at room temperature, and then 6 g (0.015 mol) of the bromide 2 are added, and the mixture is refluxed for 3 days. An oil is obtained by working up, which on a column with 500 g of silicon dioxide gel is cleaned. 5.5 g (90%) of the alkyne are obtained in the form of an oil which is sufficiently pure for the following stages is.

6ß-methoxy-25-methyl-3Q ^, 5-cyclo-5o6-cholestane (11)

A mixture of 5.1 g (0.13 moles) of 10.75 ml of dioxane, 1.0 g of sodium bicarbonate and 0.1 g of 10% Pd / C is H-stirred under 1.01 bar (1 atm), until 2 equivalents of gas are consumed. The solids are then removed by filtration and the filtrate is concentrated in vacuo to give an oil which after purification on a column of 400 g of silica gel yields 4.75 g (90%) of the 25-methyl derivative of 11 leads.

Solvolysis of compound 12: formation of 25-methylcholesterol- 3ß-acetate (12)

2 mmol of 3,5-cholesteroid 12, dissolved in 50 ml of glacial acetic acid, become 18

Heated to 70 ° for hours. After cooling, the mixture becomes with 10% aqueous NaOH and neutralized three times with 75 ml each Ethyl acetate extracted. The organic extracts are combined and washed three times with 50 ml of 10% aqueous NaOH, then with saturated Saline and washed with water. Evaporation of the organic solvent gives compound 12 as amorphous pesticide in 85-90% yield with sufficient purity for the following reaction stages.

3ß-acetoxy-25-methylcholesta-5,7-diene (13)

1 mmol of cholesterol derivative 12 in 40 ml of CCl ₄ is treated with 600 mg of NaHCO ₃ and 170 mg of 1,3-dibromo-5,5-dimethylhydantoin, and the mixture is heated _{under reflux under N 3 for 3 hours.} After cooling to 0 ° C., the solid hydantoin is removed by filtration. The filtrate is evaporated and the residue is redissolved in 5 ml of xylene at 140 ° C. After 1.5 hours under N ₂ at this temperature, the mixture is cooled to room temperature, diluted with benzene and _{washed with 5% HCl, 4% NaHCO 3} and saturated NaCl solution. After drying (Na ₂ SO ₄ ), the solvent is evaporated and the residue is _{chromatographed on AgNO 3} -impregnated thin-layer plates, developed with ethyl acetate / Skellysolve B (1: 4). This leads to pure 5,7-diene (13) with a molecular weight of 440 and the typical 5,7-diene ultraviolet spectrum.

25-methyl-vitamin D. ₃ (14)

0.02 mmol of the 5,7-diene 13 in 100 ml of diethyl ether are _{irradiated under N 2} and in an ice bath for 3 minutes with vigorous stirring, using a water-cooled quartz irradiation device and a low-pressure mercury arc lamp with a Vycor filter . The solvent is then evaporated and

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the residue is purified by preparative thin layer chromatography (ethyl acetate / hexane). The previtamin derivative is obtained and subjected to hydrolysis in 0.1 M KOH / MeOH at 70 ° for 3 hours. This step completes both the removal of acetate and the isomerization of the previtamin skeleton to form the vitamin compound. The basic medium is _{diluted with H 3} O and extracted with ethyl acetate. After the organic solvent has evaporated, the product is purified on silica gel plates impregnated with AgNO (ethyl acetate / Skellysolve B). This leads to the vitamin analog, 25-methyl-vitamin D ^ (compound 14), in pure form with the expected physical properties (molecular weight 398; ultraviolet maximum 265 nm).

Cholanocalciferol can be produced according to the following reaction scheme:

OTs

85% ν

7054

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AcO

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7-dehydrocholenic acid dimethylamide-3ß-acetate (8)

A mixture of 500 mg of the amide 4, 45 ml of carbon tetrachloride, 665 mg of sodium bicarbonate and 194 mg of 1,3-dibromo-5,5-dimethylhydantoin is refluxed _{under N 2 for 3 hours.} After cooling to 0 C, the precipitated hydantoin is removed by filtration. The filtrate is evaporated, redissolved in 5 ml of xylene and added dropwise to a mixture of 300 ml of collidine in 65 ml of xylene at 140 ° C. The reaction is kept at this temperature for 1.5 hours under nitrogen, cooled to room temperature, diluted with 100 ml of benzene, washed with dilute HCl, then with NaHCO ^ solution and finally with saturated salt solution. The product is then chromatographed on AgNO Ultraviolet spectrum for a 5 ", 7-diene chromophore.

Irradiation of compound 8: formation of vitamin analog 9

0.025 mmol of diene 8, dissolved in 100 ml of diethyl ether, and cooled in an ice bath, are _{irradiated for 3 minutes under N 2} and vigorous stirring using a water-cooled quartz irradiation device and a low-pressure mercury arc lamp with a Vycor filter. The solvent is evaporated and the residue is purified by preparative thin-layer chromatography on silica gel. The previtamin derivative is obtained and subjected to hydrolysis (0.1 m KOH / MeOH, 50-70 °, 3 hours) in order to carry out both the hydrolysis of the acetate and the thermal isomerization of the previtamin to form the vitamin structure. The hydrolysis mixture is then diluted with H "0, - extracted with ethyl acetate and after evaporation of the solvent, the resulting product is cleaned on silicon dioxide gel thin-layer plates, developed with ethyl

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acetate / Skellysolve B. This leads to (30%) the amide vitamin analog 9 (Cholanocalciferol-dimethylamide) in pure form, with the expected physical properties, i.e. H. Molecular weight 399 and ultraviolet maximum at 265 nm.

Synthesis of cholanocalciferol dimethylamide (vitamin analog 9)

As shown in process scheme 1, the cholenic acid dimethylamide derivative 4 is also used as an intermediate for the preparation of the
5,7-diene-dimethylamide 8. The conversion of 4 to 8 is achieved in the usual way, namely by allylic bromination and
Dehydrobromination. Purification of the desired 5,7-diene 8
is carried out by chromatography on AgNO ^ -impregnated silicon dioxide. By irradiating 8 in ether (in exactly the same way as described for the conversion of 6 into 7), the previtamin derivative is obtained, which after thermal isomerization, saponification (0.1 m KOH / MeOH / 70 ° / 3 hours ) and purification (preparative silica gel thin layer plates, using 25% ethyl acetate in Skellysolve B) das
Amide-vitamin 9 results in a homogeneous state.

The amide 4 used as starting material was obtained as described in the above literature references or in the main patent. The bromide 2 was reacted with n-butyllithium and dimethylacetamide in tetrahydrofuran to form the cyclosteroid 3, which was heated in glacial acetic acid at 70 ° C. for 18 hours and then neutralized to form the desired amide
became.

The intermediates 8 and 13 form a further subject of the invention. You can under basic conditions in the usual Wise hydrolyzed, with the formation of 7-dehydrocholenic acid-dimethylamide or of 3ß-hydroxy-25-methylcholesta-5,7-diene.

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The invention also relates to the use of cholancalciferol dimethylamide or 5-methyl-vitamin D ₃ for the treatment of calcium diseases or calcium disorders in animals.

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Claims (1)

19.609 - hr PATENT LAWYERS _{Λ Λ} ^ _{Λ Λ Λ} Dr. rcr. nat. D-ETER LOUIS 2920092 PipL-Phys. CLAUS FÖHLAU
DipL-Ing. FRANZ LOKRSNtTZ
Θ500 NORNBERQ KESSUERfLATZ 1 WISCONSIN ALUMNI RESEARCH FOUNDATION, North Walnut Street, Madison, Wisconsin 53707, V.St.A. Patent claims: Cholanocalciferol - dimethylamide. 25-methyl-vitamin D ₃ , 7-dehydrocholenic acid-dimethylamide-3ß-acetate, 3ß-acetoxy-25-itiethylcholesta-5,7-diene, 7-dehydrocholenic acid dimethylamide, 3β-Hydroxy-25-methylcholesta-5,7-diene as a further training of the connections of patent (patent application P 28 12 741.5). Pharmaceutical composition containing cholanocalciferol-dimethylamide and / or 25-methyl-vitamin D ₃ , optionally together with a pharmaceutically acceptable carrier or diluent. Use of cholanocalciferol-dimethylamide or 5-methyl-vitamin D-, used to treat calcium disorders Animals. 909848/0701