JPS6228784B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to compounds characterized by anti-vitamin D activity. More specifically, the present invention provides anti-vitamin D
Concerning vitamin D analogues that exhibit activity. Still more particularly, the present invention relates to vitamin D analogs that, when administered together with vitamin D to an animal, prevent or eliminate the animal's normal response to vitamin D. The use of various vitamin Ds to correct calcium metabolism disorders, such as rickets, has been known for a long time. More recently, various vitamin D derivatives, such as 25-hydroxy cholecalciferol (U.S. Pat. No. 3,565,924), 1α-hydroxycholecalciferol (U.S. Pat.
No. 3,741,966) and 1,25-dihydroxycholecalciferol (US Pat. No. 3,697,559) have made it possible to treat metabolic disorders, including calcium and phosphorus imbalances, in animals (e.g. , U.S. Patent No. 3646203
No. 3639596 and No. 3879548). Other vitamin D derivatives have now been discovered that are characterized by anti-vitamin D activity. In other words, such compounds, when administered together with vitamin D, act to abolish the animal's normal response to vitamin D. That is, these compounds contain vitamin D
It shows the effect of blocking the intestinal calcium transport effect and bone calcium movement effect. These activities suggest that these compounds can be used to correct calcium disorders such as hypercalcemia, hypervitaminosis D, vitamin D hypersensitivity, and metastatic calcification in animals. A preferred compound of the invention has the following structural formula. In the formula, X is as well as R ₁ is lower alkyl, R ₂ and R ₃ are each hydrogen, lower acyl, lower alkyl or aryl. The terms "lower alkyl" and "lower acyl" refer to alkyl and acyl substituents containing from 1 to about 6 carbon atoms. Examples include methyl, ethyl, propyl, etc., and acetyl, propionyl, etc., respectively. Suitable aryl substituents are phenyl or substituted phenyl groups. It is to be understood that R ₂ and R ₃ are each the same or different of the above substituents. It is generally known that esterified products of vitamin D are easily hydrolyzed into free vitamins in animal bodies by esterases (Sebrell et al.
& Harris “The Vitamins” Vol.
Academie Press NY (1954), P. 143), esterified products of the above compounds are also included within the scope of the present invention. Such compounds are represented by the following general formula. In the formula, X is as defined above, and R is an acyl group having 1 to about 6 carbon atoms or benzoyl. These compounds may include specific vitamin D derivatives, i.e. compounds of the above formula (where R is hydrogen), in combination with a suitable acid chloride or acid anhydride, such as acetyl chloride if the acetate ester of this specific compound is desired. Alternatively, it can be easily produced by reacting it with acetic anhydride in the presence of an organic base. The biological activity of vitamin D, especially vitamins D ₂ and D ₃ , is due to the conversion to 25-hydroxylated form, which is an intermediate that is always produced during the metabolism of vitamin D in the animal body. has been widely proven. Therefore, in vivo,
An agent that inhibits the hydroxylation of vitamin D at the C-25 position of the molecule renders vitamin D biologically inactive to perform its well-known vitamin D function. That's what happens. Thus, the biological properties of the compounds of the invention, namely their anti-vitamin D properties, are due to the fact that the substituent shown at the C-25 position in the molecule (formally, the position corresponding to the C-25 position) is 25- This is believed to be due to the fact that vitamin D is not hydroxylated by the enzyme hydroxylase (which normally converts vitamin D to 25-hydroxyvitamin D in the animal body). It is not subject to any restrictions. Hereinafter, the production of the vitamin D analog of the present invention will be described, but this is merely an example of the present invention, and the present invention is not limited thereby. In the following explanation and reaction scheme regarding the production of the compounds of the present invention, specific compounds are indicated by numbers. First, 25-methylvitamin D ₃ (in the general formula representing the compound of the present invention, X is ) can be produced according to the following reaction system. Next, this reaction system formula will be explained. Halides such as bromides in tetrahydrofuran
(2) and the lithium salt of 3,3-dimethylbutyl (n-
Acetylenic intermediate (3) is obtained by reaction with (produced with butyllithium). Compound (3) was hydrogenated (H ₂ , 10% pd/C) to give 25-methylcholesterol 3β- by solvolysis in hot glacial acetic acid.
Compound (4) is obtained which converts into acetate [compound (5) ]. This intermediate acid was brominated at the site corresponding to the allyl group and then dehydrobrominated to obtain 5,7-diene (6) , and compound (6) (3β-acetoxy-25methylcholesterol-5.7-diene). After irradiating the light,
Subsequently, the obtained previtamin derivative is subjected to thermal isomerization and saponified with a mixture of KoH/MeoH to obtain 25-methylvitamin D ₃ (7) . This method for producing 25-methylvitamin D ₃ will be explained in more detail based on Examples. (20-S)-6β-methoxy-20(p-toluenesulfonoxymethyl)-3α,5-cyclo-5
-Synthesis of α-pregnan (1) The starting material (1) was prepared by JJ Partride, S. Faber, MR.
Uskokovic's method (Helu.Chim.Acta 57 , 764
(1974)), 61% from stigmasterol
was produced with an overall yield of . It had the following physical properties. mp144−145℃; [α] ²⁰ _D = +34
(C98.CHcl ₃ ); 1R (Ccl ₄ ) 1190 and 1180 cm ^-1 (sulfonate), 1100 cm ^-1 (i-ether C-
O); NMR (CDcl ₃ ) δ7.79 and 7.34 (AB, 4H, J
_AB =8Hz. tosylate) 3.98 (d of d, J ₁ =
9.0, J ₂ = 3.4Hz. 1H, 22ε ₁ ), 3.78 (d of d, J ₁
=9.0, _J2 =5.9Hz, 1H, _22ε2 ), 3.31(S,
3H, O-Me), 2.76 (d of d, J ₁ = J ₂ = 2.7Hz.
1H, 6α-H), 2.45 (S.3H. tosylate). 1.01
(S.3H, C-19),. 98(d, J=6Hz, 3H.C−
twenty one). ．． 68 (S, 3H, C-18),. 42 (d of d, J ₁
=7.8, J ₂ =4.8Hz, 1H, C-4α; mass spectrum m/e (relative intensity) 500 (M ⁺ , 19), 485(12),
468 (68), 445 (21), 296 (45), 281 (28), 91
(100); homogeneous on TLC (Rf = .50, 20% ethyl acetate in Skellysolve B). Synthesis of (20-S)-20-promomethyl-6β-methoxy-3α-5-cyclo-5α-pregnane (2) 2.30 g tosylate in 314 ml acetonitrile
(1) 3.51 g of dry, powdered lithium bromide under reflux 6.5 g
heated for an hour. After removing the solvent by flash evaporation, the solid was dissolved in 150 ml of water and washed three times each time.
Extracted with 50ml dichloromethane. The mixed extract is
It was washed twice with 50 ml of H ₂ O and once with 50 ml of saturated salt, dried over sodium sulfate and evaporated to a flash. By crystallization from aqueous ethanol solution,
1.88g (85%) of compound (2) was obtained. mp63.5
−65°C; [α] ²⁰ _D = +55° (C1.60, CHcl ₃ ); 1R
(Ccl ₄ ) 1100cm ^-1 (C-O of i-ether);
NMR (CDcl ₃ ) δ3.52 (d of d, J ₁ = 9.8, J ₂ = 2.8
Hz, 1H, C-22ε ₁ ) 3.34 (d of d, J ₁ = 9.8,
_J2 = 5.4Hz, 1H, C- _22ε2 ), 3.32(S, 3H,
O-Me), 2.77 (d of d, J ₁ = J ₂ = 2.9Hz, 1H,
C-6α), 1.09 (d, J=6.3Hz, 3H, C-
21), 1.02 (S, 3H, C-19), . 75 (S, 3H, C
-18),. 68 (d of d, J ₁ = 7.9, J ₂ = 4.0Hz, 1H,
C-4β). ．． 43 (d of d, J ₁ = 7.9, J ₂ = 5.0Hz,
1H, C-4α); Mass spectrum m/e (relative intensity) 410 (52), 408 (48), 395 (40), 393
(42), 378 (98), 376 (100), 355 (78), 353
(85); Uniform on TLC (Rf=.70, Skerisolve B
10% ethyl acetate); 98% purity by GLC (tr=
4.1min); Elemental analysis, calculated value of C ₂₃ H ₃₇ OBr C ₁ 67.47; H.9.11.
Actual value C, 67.49; H, 9.23. Synthesis of 6β-methoxy-25-methyl-3α,5-cyclo-5α-cholesto-26-yne (3) 25 ml of 1.5M hexane solution of n-butyllithium
was added at 5°C to a solution of 3g of 3,3-dimethyl-1-butyl dissolved in 120ml of dioxane. After stirring the mixture at room temperature for 4 hours, the bromide
6 g (0.015 mol) of 2 was added and the mixture was refluxed for 3 days. As a result, an oil was obtained and purified on a column of silica gel (500 g). 5.5 g (90%) of alkyne (3) were thus obtained in the form of an oil, with sufficient purity to be used in the next step. Synthesis of 6β-methoxy-25-methyl-3α,5-cyclo-5α-cholestane (4) Compound (3) 5.1g (0.013mol), dioxane 75ml,
A mixture of 1.0 g of sodium bicarbonate and 0.1 g of 10% PD/C was stirred in 1 atm of hydrogen gas until 2 equivalents of gas were consumed. The solid material was removed by filtration and the filtrate was concentrated in vacuo to give an oil. This oil was purified using a silica gel (400g) column and
-Methyl derivative (4) (4.75 g, 90%) was obtained. Solvolysis of compound (5) (preparation of 25-methylcholesterol 3β-acetate (5) 2 mmol of 3,5-cyclosteroid (5) was dissolved in glacial acetic acid.
It was dissolved in 50 ml and heated at 70°C for 18 hours. After cooling the solution, it was neutralized with 10% NaOH aqueous solution and extracted with 75 ml of ethyl acetate, and this extraction was repeated three times. The organic extracts were collected and diluted with 10% NaOH aqueous solution (3 x 50
ml), and then further washed with a saturated salt solution and water. When the organic solvent was evaporated, compound (5) was obtained as an amorphous solid in a yield of 85-90%. This was of sufficient purity to be used in the next step. 3β-acetoxy-25-methylcholesteria
Synthesis of 5,7-diene (6) Cholesterol derivative (5) 1 mmol to 40 ml Ccl ₄
600 mg of NaHCO ₃ and 170 mg of
It was treated with 1,3-dibromo-5,5-dimethylhydantoin and the mixture was refluxed under nitrogen for 3 hours. After cooling to 0° C., the solid hydantoin was removed by filtration. The filtrate was evaporated and the residue was redissolved in 5 ml of xylene at 140°C. After standing at this temperature in a nitrogen atmosphere for 1.5 hours, the mixture was cooled to room temperature, diluted with benzene, and then
% HCl, 4% NaHCO ₃ and saturated NaCl solution. After drying with Na ₂ SO ₄ the solvent was evaporated and the residue was washed with ethyl acetate/Skerisolve B.
Chromatography was carried out on AgNO ₃ -impregnated thin plates using (1:4) as developer. It was confirmed that pure 5,7-diene (6) with a molecular weight of 440 and a typical ultraviolet spectrum of 5,7-diene was produced. Synthesis of 25-methylvitamin D ₃ (7 ) 0.02 mmol of 5,7-diene (6) was dissolved in 100 ml of diethyl ether and stirred vigorously using a water-cooled quartz light irradiation device and a low-pressure mercury arc lamp equipped with a Vicol filter. Light irradiation was carried out for 3 minutes on an ice bath in a nitrogen atmosphere. The solvent was then evaporated and the residue purified by preparative thin layer chromatography (ethyl acetate/hexane). In this way, the previtamin derivative was extracted. This previtamin derivative was prepared using 0.1M KOH/MeOH.
Hydrolysis treatment was carried out at 70°C for over 3 hours. This step achieved removal of acetate and isomerization of the previtamin backbone into vitamin compounds. The basic medium was diluted with _H2O and extracted with ethyl acetate. After evaporating the organic solvent, the product was converted into AgNO ₃
Purification was carried out on impregnated silica gel plates (developing agent: ethyl acetate/Skerisolve B). This allows the vitamin analogue, 25-methyl-vitamin D ₃
(Compound (7) ) was obtained in pure form. The molecular weight of this product was 398, and the maximum ultraviolet wavelength was 265 nm. 3β-hydroxy-25-methylcholester-5.
Preparation of 7-diene The 3β-acetoxy-25-methylcholestear-5,7-diene (6) obtained above was hydrolyzed under basic conditions according to a conventional method, and the yield was %.
Hydroxy-25-methylcholester-5,7-diene was obtained. Next, cholanocalciferol dimethylamide (in the general formula representing the compound of the present invention described above, X is The method for producing (in the case of ) can be expressed by the following reaction system formula. Next, the above reaction system formula will be explained. (20-S)-6β-methoxy-20(ρ-toluenesulfonoxymethyl)-3α,5-cyclo-5
halogenating α-pregnane (1) with a reactant selected from the group consisting of lithium bromide and lithium iodide and condensing the resulting halogenated product (2) with dimethylacetamide in the presence of a strong base; The corresponding choleic acid dimethylamide (8) is prepared and the resulting condensation product is heated with glacial acetic acid to produce the corresponding acetylated choleic acid dimethylamide (9) . Next, compound (9) can be converted to compound (10) according to a conventional method. namely, allyl bromination and dehydrobromination. Purification of the target 5,7-diene (10) is achieved by chromatography on AgNO ₃ -impregnated silica gel. Light irradiation of compound (10) in ether gives a previtamin derivative, which undergoes thermal isomerization and saponification (0.1M KOH/MeOH/70°C/
3 hr) and purification (on silica gel separation plates, employing 25% ethyl acetate in Skellisolve B) to give the amide vitamin (11) in a homogeneous state. The method for producing cholanocalciferol dimethylamide will be explained in more detail based on Examples. Synthesis of 6β-methoxy-3α,5-cyclo-5α-cholanic acid dimethylamide (8) A dry flask filled with 4.9 ml of 1.79Mn-butyllithium and 14.5 ml of tetrahydrofuran, 3.6 ml of hexane, and 1.55 ml of diisopropylamine. added to. The mixture was kept at 0°C for 0.5 h under a nitrogen atmosphere, then the flask was cooled to -78°C, 1.63 ml of dimethylacetamide was added, and the mixture was heated to -78°C for 1.25 h.
Stir for hours. The bromide (compound (2) , 1.2 g in 15 ml tetrahydrofuran) was added and the mixture was heated to 0°C.
The mixture was stirred for 21 hours. After adding a few ice cubes followed by 50 ml of 5% HCl, the mixture was diluted with dichloromethane for 3
Extracted twice (50 ml for each extraction). The organic extracts were combined, washed with saturated salt, dried over sodium sulfate, and flash evaporated. The residue is silica gel
Chromatographed on a 250g column. 0.85g (70%) of compound (8) when eluted with ethyl acetate
(Pooltube 34-67, 15ml distillate)
was obtained as a clear oil. This oil did not crystallize. The properties of this oil were as follows. IR (Ccl ₄ ) 1655cm ^-1 (amide), 1100cm ^-1 (i-
Ether C-O); NMR (CDcl ₃ ) δ3.32
(S, 3H, O-Me), 3.00 (S, 3H, N-Me),
2.93 (S, 3H, N-Me), 2.77 (d of d, J ₁ = J ₂
=2.9Hz, 1H, C-6α), 1.02(S, 3H, C-
19),. 94 (d, J=5.9Hz, 3H, C-21),. 72
(S, 3H, C-18),. 43 (d of d, J ₁ = 8.3, J ₂ =
4.8Hz, 1H, C-4α); Mass spectrum m/e
(Relative intensity) 415 (M ⁺ , 6), 400 (7), 383 (26),
368(8), 360(13), 100(32), 87(100); High resolution mass spectrum, calculated value of C ₂₇ H ₉₅ NO ₂ ,
415.345, actual value 415.343. Synthesis of 3β-acetoxy-5-choleic acid dimethylamide (9) 1.86 g of cyclosteroid (8) was dissolved in 75 ml of glacial acetic acid and the mixture was then heated to 70° C. for 18 hours. After cooling to room temperature, the mixture was neutralized with a 10% aqueous solution of sodium hydroxide and diluted with ethyl acetate.
Extracted twice (100 ml for each extraction). The organic extracts were combined and washed three times with a 10% aqueous solution of sodium hydroxide (50 ml for each wash), once with 50 ml of water, and once with 50 ml of saturated salt.
Washed twice. Flash evaporation gave an amorphous solid, which was crystallized from hexane to yield 1.85 g.
(93%) of compound (9) was obtained. The properties of this compound were as follows. mp192−193.4℃;
[α] ²⁰ _D = -41° (C1.1, CHcl ₃ ); IR (Ccl ₄ ) 173
3
and 1245cm ^-1 (acetate), 1651cm ^-1 (amide); NMR (CDcl ₃ ) δ5.37 (m, 1H, C-6),
4.62 (m, W1/2=20Hz, 1H, C-3α), 3.00 (S, 3H, N-Me), 2.93 (S, 3H, N-Me),
2.03 (S.3H, acetate), 1.02 (S, 3H, C-
19),. 95 (d, J=5.9Hz, 3H, C-21),. 68
(S, 3H, C-18); Mass spectrum m/e (relative intensity) 443 (M ⁺ , .8), 428 (.6), 383
(65), 368(6), 100(70), 87(100); Homogeneous on TLC (Rf = .55, ethyl acetate); 96% purity on GLC (tr = 28.6 min); Elemental analysis, C ₂₈ H ₄₅ NO ₃ calculated value C, 75.80; H, 10.22; N, 3.16, actual value C,
75.70; H, 10.30; N, 3.09. 7-dehydrocholeic acid dimethylamide 3β-
Synthesis of acetate (10) Amide (9) 500 mg, carbon tetrachloride 45
ml, 665 ml of bicarbonate of soda and 1,3-dibromo-
The mixture consisting of 5,5-dimethylhydantoin was refluxed under nitrogen for 3 hours. Then cooled to 0°C,
The precipitated hydantoin was removed by filtration. After evaporating the filtrate, redissolve it in 5 ml of xylene.
A mixture of 300 ml of collidine and 65 ml of xylene was added dropwise at 140°C. The reaction was carried out at this temperature for 1.5 hours under nitrogen and after cooling to room temperature was diluted with 100 ml of benzene and washed with dilute hydrochloric acid, then with NaHCO ₃ solution and saturated NaCl solution. The product was converted to ethyl acetate/Skerisolve B.
Chromatography was performed on a thin silica gel plate impregnated with AgNO ₃ using as a developing agent. Pure 5,7-diene (compound (10) ) was obtained with a yield of 20%. This product had a molecular weight of 441 and exhibited a typical ultraviolet spectrum of 5.7-diene chromophore. Light irradiation of compound (10) : synthesis of vitamin analogs 0.025 mmol of diene (10) was dissolved in 100 ml of diethyl ether, cooled in an ice bath, and dissolved in nitrogen for 30 minutes.
Light was irradiated for minutes with vigorous stirring using a water-cooled quartz light irradiation device and a low-pressure mercury arc lamp equipped with a Vicol filter. The solvent was then evaporated and the residue purified by preparative silica gel thin layer chromatography. Collect this previtamin derivative and add 50% with 0.1M KOH/MeOH.
Hydrolysis was carried out at ~70°C for 3 hours, and the vitamin structure was obtained by hydrolysis of the acetate ester and thermal isomerization of the previtamin. After diluting the hydrolysis mixture with water, extracting with ethyl acetate, and evaporating the solvent, the resulting reaction product was deposited on a silica gel plate.
It was developed and purified using ethyl acetate/Skellysolve B. This gave the amide vitamin analog (11) (cholanocalciferol dimethylamide) in pure form (30%). It has the expected physical properties, a molecular weight of 399,
The maximum ultraviolet wavelength was 265 nm. Preparation of 7-dehydrocoleic acid dimethylamide 7-dehydrocoleic acid dimethylamide 3β-acetate (10) obtained above was hydrolyzed under basic conditions according to a conventional method, and the yield was %. Inic acid dimethylamide was obtained.

Claims (1)

[Claims] 1. General formula (X in the formula is as well as R is selected from the group consisting of hydrogen, an acyl group having 1 to about 6 carbon atoms, and benzoyl, R ₁ is lower alkyl, R ₂ and R ₃ are each hydrogen,
Lower alkyl, lower acyl or aryl. ) A compound represented by 2. The compound according to claim 1, which is 225-methylvitamin _D3 . 3. The compound according to claim 1, which is cholanocalciferol dimethylamide. 4 (20-S)-6β-methoxy-20(ρ-toluenesulfonoxymethyl)-3α,5-cyclo-
5α-pregnane is halogenated with a compound selected from the group consisting of lithium bromide and lithium iodide, and the resulting halogenated product is reacted with the lithium salt of 3,3-dimethylbutyne to form 6β-methoxy-25- Methyl-3α,5-cyclo-5α-cholesto-23-yne was prepared and hydrogenated to give 6β
-methoxy-25-methyl-3α,5-cyclo-5
α-Cholestane was obtained and the compound was solvolyzed in glacial acetic acid to obtain 25-methylcholesterol 3β.
-acetate, this compound is brominated and then dehydrobrominated to obtain 3β-acetoxy-25 methylcholesterol-5,7-diene, which is irradiated with light, the obtained previtamin derivative is isomerized, and then saponified. A method for producing 25-methylvitamin _D3 . 5 (20-S)-6β-methoxy-20(ρ-toluenesulfonoxymethyl)-3α,5-cyclo-
5α-pregnane is halogenated with a reactant selected from the group consisting of lithium bromide and lithium iodide, and the resulting halogenated product is condensed with dimethylacetamide in the presence of a strong base to form the corresponding choleic acid dimethylamide. and heating the resulting condensation product with glacial acetic acid to form the corresponding acetylated choleic acid dimethylamide, which is brominated and then dehydrobrominated to give 7-
Dehydrocholenic acid dimethylamide 3β
- A method for producing cholanocalciferol dimethylamide, which comprises obtaining acetate, irradiating it with light, and then isomerizing and saponifying the obtained previtamin derivative.