JP2004277433A - Vitamin d compound and method for producing the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new synthetic intermediate for synthesizing vitamin D compounds. <P>SOLUTION: The intermadiate is hydrindan-4-one compound represented by formula (IV) or (V). Wherein, R<SB>2</SB>is a 1-3C alkyl, a hydroxy 1-3C alkyl, a 1-2C alkoxymethyl or a 2-3C alkenyl or alkynyl, R<SB>3</SB>is a bivalent (branched) and (saturated) aliphatic 3- to 5-membered hydrocarbon or oxahydrocarbon which have at least 3 atoms in the main chain and optionally substituted with one or more substituents selected from epoxy, fluoro and hydroxy, R<SB>4</SB>is a sec- or tert- 1-3C alkyl or a 3-6C cycloalkyl, and R<SB>5</SB>is (protected) hydroxy. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to novel vitamin D compounds, processes for preparing these compounds and their use in medicine and cosmetics.

  It is generally known that vitamin-D compounds or vitamin-D related compounds ("vitamin-D compounds") have strong biological activity and can be used in all cases involving calcium metabolism problems. Have been. A few years ago, various active vitamin-D compounds also have other pharmacological activities and are used, for example, for the treatment of certain skin and bone diseases, for cosmetic use, and for diabetes mellitus. It has been found that it can be used successfully for the treatment of hypertension and diseases associated with cell differentiation, cell proliferation or an imbalance in the immune system, including inflammatory diseases such as rheumatoid arthritis and asthma. In addition, these compounds can be used in various animal applications and for diagnostic purposes.

Vitamin D compounds of interest for the above applications are hydroxylated vitamin D compounds, especially those hydroxylated at the 1α-, 24- and / or 25-position. A recent development in the field of active vitamin D compounds is that 19-nor-vitamin D compounds, which are preferably also hydroxylated in the 1α-position and optionally in the C ₁₇ -side chain (US Pat. And C ₁₈ -modified vitamin D compounds (Patent Document 2). C _{17 -} also other modifications of the side chains have been proposed in order to suppress and adverse side effects for improving the activity intended similarly. Examples of C ₁₇ -side chain modifications include chain extension (homocompounds), 22-oxa modification, fluoro substitution, epoxy group (eg, Patent Document 3). In addition, certain 24-cyclopropyl-modified vitamin D compounds have been disclosed in the literature, such as, for example, US Pat. However, in general, the above C ₁₇ -side chain modified vitamin D compounds are still not completely satisfactory with respect to their selective activity, ie the intended activity without deleterious side effects.

Moreover, the availability of C ₁₇ -side chain modified vitamin D compounds is often inadequate or uninteresting. For example, the preparation of the above vitamin D compounds disclosed by Farachi-Carson et al. Appears to be very labor intensive, and the C ₁₇ -side chain formation described in US Pat. There is a need for various laborious synthetic steps using ketones that are not available. If directed to this, better availability of C _{17 -} there is a need for a side chain modified vitamin D compounds. Indeed, the starting compounds for the preparation of such vitamin-D compounds must be readily available or available, and it is contemplated that the multi-step preparation method will have sufficient selectivity and effect. You have to achieve your purpose.
EP-A-0387077 EP-A-0521550 WO 92/21695 WO 87/00834 Farach-Carson et al. , Endocrinology, 1991, 129, 1876-84.

  Accordingly, it is an object of the present invention to provide a new class of vitamin D compounds that are readily available or that are well obtained from available starting materials.

  According to the present invention, this object is represented by the general formula

(In the formula,
R ₁ is a hydrogen atom or a hydroxy group,
R ₂ represents a (C ₁ -C ₃ ) alkyl group, a hydroxy (C ₁ -C ₃ ) alkyl group, (C _1-
C ₂₎ alkoxymethyl group or _(C 2 -C ₃₎ alkenyl or alkynyl group,
R ₃ if and has at least 3 atoms with an epoxy, one or more of which do also be straight-chain or min substituted with a substituent selected from the full Oro and hydroxy in the main chain A branched, saturated or unsaturated aliphatic 3- to 5-membered hydrocarbon or oxahydrocarbon divalent group;
R ₄ is a sec- or tert- (C ₃ -C ₆ ) alkyl group or a (C ₃ -C ₆ ) cycloalkyl group, and A and B are each independently a hydrogen atom or a methyl group, or A and B together form a methylene group.
New vitamin D compounds.

  The novel vitamin D compounds of the present invention represented by the general formula I are valuable substances. Biological outcomes, as exemplified in the examples, show that these compounds are promising as biologically active substances and, in all the above pharmacotherapy indications, more particularly osteoporosis, Renal osteodystrophy, osteomalacia, skin diseases such as psoriasis (and other hyperproliferative skin diseases), eczema and dermatitis, myopathy, leukemia, breast and colon cancer, osteosarcoma, squamous cell carcinoma, melanoma It has been shown that it can be used for the treatment of certain immunological diseases, as well as transplant rejection.

  In addition, the novel vitamin D compounds of the present invention can also be used for wound healing and to prevent, regulate and / or protect the skin as well as, for example, wrinkles, dry skin, sagging and insufficient skin It can be incorporated into cosmetic compositions such as creams, lotions, ointments and the like to improve various skin conditions such as sebum secretion. The novel vitamin D compounds can also be used for diagnostic purposes.

Suitable examples of the above substituents _{R 4} is isopropyl, cyclopropyl, tert - butyl, thexyl (1,1,2-trimethyl-propyl), 3-pentyl and cyclopentyl.

R ₁ is a hydroxy group,
R ₂ has the above meaning,
R ₃ has the formula _{-O-CH 2 - (CH 2} ) n -, - CH 2 -CH 2 - (CH 2) n-,
-CH = CH- _(CH 2) n- or _{-CH 2 -CH 2 -CH (CH 3} ) -
Is a divalent group of
Where n is 1 or 2,
R ₄ is an isopropyl, cyclopropyl or tert-butyl group, and A and B are hydrogen atoms or together form a methylene group;
Preference is given to vitamin D compounds having the above general formula I.

Examples of very suitable vitamin D compounds according to the invention because of their very favorable biological properties are the meanings where the symbols R ₁ , R ₂ , R ₃ , A and B are defined above. And wherein R ₄ is an isopropyl group.

A particular advantage of the present invention is that the above described novel vitamin D compounds of the present invention can be easily prepared from readily available starting materials. In particular, it has been found that the desired C ₂₅ -structure, ie the attachment of a suitable substituent to C ₂₅ , is readily obtained by starting from readily available ester compounds.

Accordingly, the present invention also relates to a process for the preparation of a vitamin D compound of the general formula I as defined above wherein R ₁ is a hydroxy group, the process according to the present invention being of the general formula

(In the formula,
R ₂ , R ₃ , A and B have the meanings defined above,
R ₅ is a protected hydroxy group, and R ₆ is _(C 1 -C ₆₎ alkyl group]
An ester compound of the general formula
R ₄ M (X) p (III)
(In the formula,
R ₄ has the above meaning,
X is Cl, Br or I;
M is a metal selected from Li and Mg, and p is 0 or 1 depending on the valency of M]
And then removing the protecting group.

In the method of equally interesting, C _{17 -} can be finished side chain first. Accordingly, the present invention provides a compound of the general formula

(In the formula,
R ₂ , R ₃ and R ₆ have the meanings given above and R ₅ ′ is an optionally protected hydroxy group.
An ester compound of the general formula
R ₄ M (X) p (III)
(In the formula,
The symbols have the above meanings.)
With an organometallic compound of the formula

When R ₅ ′ is a protected hydroxy group, the protecting group is removed and then oxidized to give a hydrindane compound having the general formula

Of the corresponding hydrindan-4-one compound of formula V and then, optionally after protection of the hydroxy group,
(A) General formula

(In the formula,
R ₁ 'is a hydrogen atom or a protected hydroxy group, and the other symbols have the meanings described above.
Using the Wittig reagent of formula (I) or (b) after enolization and derivatization of the enolic hydroxy group,

(In the formula,
The symbols have the above meanings.)
To form a compound of general formula I in which A and B together form a methylene group, followed by removal of the protecting group. It also relates to a method for the preparation of a vitamin D compound as characterized and as defined above.

  Hydroxy groups in the above intermediates or reactants can be protected by suitable esterification or reaction with an etherifying agent. Suitable esterifying agents are C2-5 alkyl chlorocarbonates or C1-4 aromatic or saturated aliphatic carboxylic acids such as, for example, benzoic acid. In order to protect the hydroxy group in the form of ethers, etherifying agents known in principle for this purpose, such as, for example, trialkylsilylimidazoles having 1 to 6 carbon atoms in the alkyl group, Suitable are halides, trialkylsilyl triflates (-trifluoromethanesulfonate), diphenylalkylsilyl halides or diphenylalkylsilyl triflates, or derivatives thereof.

  Particularly suitable for this purpose are trimethylsilyl chloride, tert-butyldimethylsilyl chloride, dimethyl- (1,1,2-trimethylpropyl) -silyl chloride, tert-butyldimethylsilyl triflate, or trimethylsilyl-imidazole. Because these etherifying agents readily react with the hydroxy group to be protected to form an ether function, which on the one hand is sufficiently stable under the conditions of the reaction or reactions, while on the other hand This is because the tertiary-butyldimethylsilyl chloride or triflate is preferable because the tertiary-butyldimethylsilyl group can be easily removed and the [hydroxyl group] can be recovered to the original hydroxy group. Very suitable as a base This is because are Idasa.

  The enol-based hydroxy group is suitably derivatized by reaction with N-phenyltriflimide to form triflate.

The starting compounds of formula II can be easily prepared from readily available materials, for example, suitable C ₁₇ as defined above _- synthesis of vitamin D compounds having a side chain is as follows :

Introduction of _{C 18} modified to the vitamin D compounds of the present invention _{(R 2)} can be done easily and as are described in the above EP-A-0521550.

  Suitable examples of the organometallic compounds of the above general formula III include lithium compounds such as isopropyllithium, cyclopropyllithium and tert-butyllithium, and Grignard reagents such as isopropylmagnesium chloride, cyclopropylmagnesium chloride and tertiary-butyl. Butylmagnesium chloride, as well as the corresponding bromides.

  The intermediate ester compound of the general formula IX represented above is novel. Therefore, the present invention also relates to this intermediate product and a method for producing this compound. General formula

(In the formula,
R ₂ , R ₃ and R ₆ have the meanings described above, and R ₇ is a derivatized hydroxy group.
An ester compound of the general formula

(In the formula,
The symbols have the above meanings.)
, Followed by hydrogenation and isomerization can easily prepare an ester compound of general formula IX wherein A and B together form a methylene group.

The reaction is preferably carried out in two reaction stages, ie the components are first reacted under the influence of an organic base such as, for example, triethylamine and in the presence of a palladium catalyst such as, for example, (PPh ₃ ) ₂ PdCl ₂ and then hydrogen The resulting product to be hydrogenated is subjected to hydrogenation using hydrogen under the influence of a suitable catalyst such as, for example, a Lindlar's catalyst (Pd on CaCO ₃ poisoned with lead), and the resulting pre- The vitamin structure is isomerized to a vitamin structure of the general formula (IX).

  Alternatively, the ester compound of general formula IX can be easily synthesized by reacting a modified Windus Grundmann ketone of general formula X or XI with a Uttig reagent as follows:

  The symbols in the above formula are as defined above.

  The intermediate hydrindan-4-one compound of the general formula V represented above is novel. Accordingly, the present invention provides that this intermediate product as well as the hydrindane compounds of the general formula IV as defined above are preferably selected from chromium-containing oxidizing agents such as pyridinium chloroformate or pyridinium dichromate and ruthenium tetroxide. It also relates to a process for producing this compound by oxidizing with an oxidizing agent.

  The intermediate hydrindane compounds of the general formula IV represented above are also novel. Accordingly, the present invention provides this intermediate by reacting this intermediate product as well as a compound of the general formula II as defined above with a metal-organic compound of the general formula III also defined above in an inert organic solvent. The present invention also relates to a method for producing a compound.

  In order to improve the applicability of the novel vitamin D compounds of the present invention for the above pharmacotherapeutic indications, the compounds are generally formulated with an effective amount of the vitamin D compound as an active ingredient. A pharmaceutical composition comprising, in addition to a pharmaceutically acceptable carrier and / or at least one pharmaceutically acceptable auxiliary substance. Such compositions may be divided into dosage units for oral, topical (dermal) or parenteral administration containing from about 0.1 μg to about 0.1 mg of active ingredient per dosage unit. it can.

  Compositions for diagnostic purposes can contain, in addition to the vitamin D compounds of the present invention, compatible nontoxic carriers and / or at least one auxiliary substance.

  The cosmetic composition may contain, in addition to the effective amount (ranging from about 0.1 μg to about 0.1 mg per dosage unit of the dosage unit form) of the vitamin D of the present invention, a cosmetically acceptable non-toxic carrier and / or Alternatively, at least one auxiliary substance can be included.

  Finally, the present invention provides a method of administering to a warm-blooded living organism a pharmaceutical composition as hereinbefore defined or effective in an amount effective for the intended purpose. Autoimmune diseases (including diabetes mellitus), acne, alopecia, skin aging (including photo-aging), imbalance of the immune system, It also relates to methods of treating and preventing a number of disease states, including inflammatory diseases such as rheumatoid arthritis and asthma, and diseases associated with abnormal cell differentiation and / or proliferation. Examples of such diseases are psoriasis and other hyperproliferative skin diseases.

  The invention also relates to the use of a pharmaceutical composition as described above for the treatment of solid, skin or blood cancers, especially breast cancer, melanoma, squamous cell carcinoma or leukemia in warm-blooded animal survivors. .

  Cosmetic compositions as defined above, especially selected from the group consisting of creams, lotions, ointments, liposomes and gels, have a number of skin conditions, such as inadequate skin firmness or texture, insufficient skin moisture, It can be used for the treatment and prevention of wrinkles and / or insufficient sebum secretion.

  The invention will be described in more detail with reference to the following individual examples.

Example I
Preparation of vitamin esters <br/> Reaction formula:

(A). Ph ₃ P (6.5g) and imidazole (4.8 g) was added to a solution medium THF (100 ml) of the diol (1) (5.0g). The suspension was cooled to −20 ° C. and I ₂ (6.28 g) was added in one portion. After stirring for 15 minutes, the reaction mixture was warmed to room temperature, stirred for another 15 minutes, cooled to 0 ° C., and poured into saturated aqueous NaHCO ₃ (50 ml). The mixture was extracted with Et ₂ O, and the extract was washed with saturated aqueous _Na 2 _S 2 _{O 3} and _{H 2} O, dried and filtered. Concentration gave a residue, which was purified by flash chromatography (8% EtOAc / hexane) to give 7.32 g of iodide (2). After crystallization from EtOAc / hexane, the product had a melting point of 51 ° C. Identification by NMR and elemental analysis.
(B). Pyridinium dichromate (8.66 g) was added to a solution of compound (2) (3.99 g) in 50 ml of CH ₂ Cl ₂ . The mixture was stirred at room temperature for 6 hours. Et ₂ O (60 ml) was added and the resulting suspension was stirred for 15 minutes and filtered. The filtrate was washed with brine, dried, filtered, and concentrated. Purification by flash chromatography (10% EtOAc / hexane) provided the desired iodoketone (3) in 3.57 g yield. Crystallization from Et ₂ O / hexane: mp 65 ° C. Identification by NMR and elemental analysis.
(C). in hexane n-BuLi (3.5 mmol, in 1.43 ml) by the addition of cooling _{(-78 ℃) i-Pr 2} NH (3.9 mmol) for solution to produce lithium diisopropylamine. After stirring for 10 minutes, the mixture was diluted with THF (4 ml), stirred at 0 ° C for 30 minutes, and cooled to -78 ° C. A solution of ketone (3) (1.0 g) in 14 ml of THF was added slowly followed by a solution of N-phenyltriflimide (1.225 g) in 4 ml of THF. The mixture was stirred at -78 C for 2 hours. After warming to 0 ° C., the reaction was stopped by the addition of a few drops of MeOH and water. Concentrated to give the crude product, which was diluted with EtOAc / hexane (30 ml), washed with brine, filtered and concentrated. The resulting residue was purified by flash chromatography (2% EtOAc / hexane) to give 1.29 g of iodotriflate (4) as a colorless oil. Identification by NMR and elemental analysis.
(D). CuI (201 mg) and Zn EtOH _/ H 2 O of (161mg) (6ml 7: 3 , has been deoxygenated) was sonicated for 5 minutes. A solution of methyl acrylate (637 μl, freshly distilled) and iodide (4) (160 mg) in EtOH / H ₂ O (1 ml 7: 3) was added continuously, and the resulting mixture was sonicated for 40 minutes. . Diluted with Et ₂ O (15 ml) and filtered to give a solution, which was washed with brine. The aqueous phase was extracted with Et 2 _{O (30ml),} drying the organic extracts were washed with filtered, and concentrated. Flash chromatography of the residue (6% EtOAc / hexane) gave 96 mg of methyl ester (5) (colorless oil). Identification by NMR and elemental analysis.
(E). The palladium catalyzed coupling between vinyl triflate (5) and enyne (6) was performed as follows:
A mixture of enine (6) (507 mg), triflate (5) (500 mg), Et ₃ N (4.85 mmol) and (Ph ₃ P) ₂ PdCl ₂ (16 mg) in 21 ml of DMF at 75 ° C. for 1 hour. Heated. The mixture was cooled to room temperature, diluted with EtOAc / hexane (50 ml 1: 3) and washed with brine. Drying, to a residue by filtration and concentration gave which was purified by flash chromatography (2-4% _Et 2 O / hexanes) 675 mg of Jienin (7) (viscous liquid). Identification by NMR.
(F) Product (7) was hydrogenated over a Lindlar catalyst as follows:
50 μl of quinoline in 10 ml of hexane (0.2 ml) was added to a solution of dienin (7) (305 mg) in 12 ml of hexane. Pre-dried Lindlar catalyst (50 mg) was added and the resulting solution was exposed to hydrogen gas at atmospheric pressure. After stirring for 8 hours, the reaction mixture was filtered and concentrated. The residue was purified by flash chromatography (1-3% _Et 2 O / hexane) to give the protected previtamin D compound of 295 mg.
Isomerization of pre-vitamin D compound to vitamin D compound (8):
The resulting previtamin D compound (295 mg) was dissolved in 15 ml of isooctane and refluxed for 5 hours in the dark. To a residue and concentrated to give the compound of 290mg (8) which was purified by flash chromatography (2-4% _Et 2 O / hexane). The product was identified by ¹ H-NMR, ¹³ C-NMR and elemental analysis.
¹ H-NMR (δ, CDCl ₃ ): 6.24 and 6.02 (d, 2H), 5.18 (m, 1H), 4.87 (m, 1H), 4.37 (m, 1H). , 4.18 (m, 1H),
3.67 (s, 3H), 0.88 (s, 18H), 0.53 (s, 3H), 0.07 (s, 12H).
¹³ C-NMR (δ, CDCl ₃ ): 173.1, 148.4, 141.0, 35.0, 132.2, 118.0, 111.2, 72.1, 67.5, 56.3. , 51.3,
46.0, 45.7, 44.8, 40.6, 35.8, 35.3, 34.4, 31.5, 28.8, 27.6, 25.8, 25.7, 23. 4, 22.6, 22.1, 21.5, 18.7, 18.1, 18.0, 14.0, 11.9, -4.8, -4.8, -4.9,- 5.2.
Elemental _{analysis: C 38 H 66 O 4 Si} 2 Calculated for: C, 70.75; H, 10.62 .
Found: C, 70.42; H, 10.43.

The following ester compounds were prepared in a corresponding manner:
General formula

Compound number R ₂ R ₃ R ₆
(9) CH ₃ (CH ₂ ) ₄ C ₂ H ₅
(10) C ₂ H ₅ (CH ₂ ) ₃ C ₂ H ₅
Compound (9): This 24-homo compound is obtained by using a homologue of compound (1) having a 1-methyl-3-hydroxypropyl side chain as a starting material in step (a) above and acrylic in step (d) Prepared by using ethyl acid as olefin.
¹ H-NMR (δ, CDCl ₃ ): 0.08 (s, 12H), 0.52 (s, 3H), 0.87 (s, 18H), 0.90 (d, 3H), 1.25 (T, 3H), 4.11 (q, 2H), 4.20 (m, 1H), 4.37 (m, 1H), 4.88 (d, 1H),
5.18 (d, 1H), 6.01 (d, 1H), 6.24 (d, 1H).
Compound (10): This 18-homo compound uses in step (a) above a homologue of compound (1) [Compound No. (64)] described in published European patent application 521550 as starting material. Thus, it was manufactured.
¹ H-NMR (δ, CDCl ₃ ): 0.87 (s, 6H), 0.88 (t, 3H),
1.01 (d, 3H), 1.25 (t, 3H), 1.98 (t, 1H), 2.25 (m, 2H), 2.44 (dd, 1H), 4.13 (q , 2H), 4.18 (m, 1H),
4.37 (m, 1H), 4.86 (s, 1H), 5.17 (s, 1H), 6.01 (d, 1H), 6.24 (d, 1H).
Example II
Production of vitamin D compounds from vitamin esters Reaction formula:

Compound (11) was prepared as follows:
In Et ₂ O (0.51 mmol, 0.602Ml in) of cyclopropyl bromide (0.51 mmol) of t-BuLi to generate the cyclopropyl lithium added to the solution. The resulting mixture is warmed to room temperature and diluted with Et ₂ O to the 2.4 ml. 1 ml of this solution was slowly added to a cooled (−78 ° C.) solution of compound (8) (50 mg) in 3 ml of Et ₂ O. The reaction mixture was spontaneously brought to -40 ° C and quenched with a few drops of water. The resulting solution was diluted with Et 2 _O, washed with brine, dried, filtered and concentrated. The concentrate was filtered through a flash chromatography column (2% Et ₂ O / hexane) to give the product (46 mg), which was dissolved in 7 ml of THF and tetrafluorofluoride in THF at room temperature in the dark. Stirred with butyl ammonium (0.36 mmol in 0.36 ml) for 24 hours. Concentrate to give a residue, which is diluted with EtOAc (20 ml), dried, filtered, concentrated and flash chromatographed (60% EtOAc / hexane) to give 23 mg of the desired compound (11) as a white solid. Provided in solid form. ¹ H-NMR (δ, CD ₂ Cl ₂ ): 6.44 and 5.99 (d, 2H), 5.27 (br-d, 1H), 4.95 (br-d, 1H), 4. 35 (m, 1H), 4.15 (m, 1H), 0.92 (d, 3H), 0.81 (m, 2H), 0.53 (s, 3H), 0.34 (m, 8H) ).
¹³ C-NMR (δ, CD ₂ Cl ₂ ): 148.6, 143.5, 133.9, 25.1, 117.6, 111.8, 71.2, 71.0, 67.2, 57 .2, 56.8, 45.8, 43.5, 43.4, 41.0, 37.1, 36.6, 29.4, 28.0, 24.0, 22.6, 20.8 , 19.0, 12.1, 0.8, -0.5.

The following vitamin D compounds were produced in a corresponding manner:
General formula

Compound number R ₂ R ₃ R ₄
(12) CH ₃ (CH ₂ ) ₃ C (CH ₃ ) ₃
(13) CH ₃ (CH ₂ ) ₃ CH (CH ₃ ) ₂
(14) C ₂ H ₅ (CH ₂ ) ₃ CH (CH ₃ ) ₂
(15) C ₂ H ₅ (CH ₂ ) ₃ cyclopropyl (16) CH ₃ (CH ₂ ) ₄ cyclopropyl (17) CH ₃ (CH ₂ ) ₄ CH (CH ₃ ) ₂
Compound (12) was prepared by using t-butyl lithium instead of cyclopropyl lithium.
Compounds (13), (14) and (17) were prepared by using isopropyl lithium instead of cyclopropyl lithium.
Compound (12): ¹ H-NMR (δ, CDCl ₃ ): 6.38 and 6.01 (d, 2H), 5.33 (m, 1H), 5.00 (m, 1H), 4.43 (M, 1H), 4.23 (m, 1H), 1.00 (s, 18H), 0.93 (d, 3H), 0.54 (s, 3H).
¹³ C-NMR (δ, CDCl ₃ ): 148.7, 143.4, 134.0, 125.0, 117.6, 111.8, 80.0, 71.1, 67.1, 60.6 , 56.9, 56.8, 46.3, 45.8, 43.4, 42.8, 40.9, 37.0, 36.5, 34.2, 29.4, 28.8, 28 0.0, 24.0, 23.3, 22.7, 19.2, 12.1.
Compound (13): ¹ H-NMR (δ, CD ₃ OD): 0.55 (s, 3H), 0.90 (m, 15H), 2.22 (dd, 1H), 2.48 (dd, 1H), 2.83 (dd,
1H), 4.09 (m, 1H), 4.31 (t, 1H), 4.86 (b ?, 1H), 5.25 (b, 1H), 6.05 (d, 1H), 6 .29 (d, 1H).
Compound (14): ¹ H-NMR (δ, CDCl ₃ ): 0.84 (t, 3H), 0.95 (m, 12H), 1.00 (d, 3H), 2.32 (dd, 1H) ), 2.60 (dd, 1H), 4.24 (m, 1H), 4.44 (m, 1H), 5.01 (b, 1H), 5.33 (b, 1H), 6.01 (D, 1H), 6.39 (d, 1H).
Compound (15): ¹ H-NMR (δ, CD ₃ OD): 0.70 to 0.40 (m, 8H), 0.76 (m, 2H), 0.91 (t, 3H), 1. 01 (d, 3H), 2.00 (m, 1H), 2.22 (dd, 1H), 2.29 (b, 1H), 2.48 (dd, 1H), 2.83 (dd, 1H) ), 4.09 (m, 1H), 4.31 (t, 1H), 4.85 (b, 1H), 5.25 (b, 1H), 6.04 (d, 1H), 6.29. (D, 1H).
Compound (16): ¹ H-NMR (δ, CD ₃ OD): 0.20-0.40 (m, 8H), 0.79 (m, 2H), 0.95 (d, 3H), 2. 25 (dd, 1H), 2.51 (dd, 1H), 2.86 (dd, 1H), 4.12 (m, 1H), 4.35 (t, 1H), 4.89 (b, 1H) ), 5.28 (b, 1H), 6.08 (d, 1H), 6.32 (d, 1H).
Compound (17): ¹ H-NMR (δ, CD ₃ OD): 0.53 (s, 3H), 0.90 (m, 12H), 2.22 (dd, 1H), 2.48 (dd, 1H), 2.83 (dd, 1H), 4.09 (m, 1H), 4.31 (t, 1H), 4.86 (b, 1H), 5.25 (b, 1H), 6. 05 (d, 1H), 6.29 (d, 1H).
Example III
Preparation of 1- (1-methyl-5-hydroxy-5,5-diisopropyl-pentyl) -hydrindanol-4 (19) Reaction formula:

(A). The starting compound (1) was converted to the corresponding iodide (2) as described in Example I (a).
(B). The resulting compound (2) was converted using ethyl acrylate as the olefin in a corresponding manner as described in Example I (d) to yield the ester compound (18).
(C). Ester compound (18) was converted to compound (19) by reaction with excess isopropyllithium in a corresponding manner as described in Example II. The product was identified by ¹ H-NMR.

The following compounds were prepared in a corresponding manner:
General formula

Compound number R ₂ R ₃ R ₄
(20) CH = CH ₂ (CH ₂ ) ₃ cyclopropyl (21) CH = CH ₂ (CH ₂ ) ₃ isopropyl (22) CH ₃ (CH ₂ ) ₄ isopropyl (23) CH ₃ (CH ₂ ) ₄ cyclopropyl The product was identified by ¹ H-NMR.
Example IV
Preparation of 1- (1-methyl-5-hydroxy-5,5-diisopropyl-pentyl) -hydrindanone-4 (24) Reaction formula:

Oxidation of compound (19) by using pyridinium dichromate as oxidizing agent in a corresponding manner as described in Example I (b) to give the desired ketone (24) in 84% yield. Given in. The product was identified by ¹ H-NMR.

The following compounds were prepared in a corresponding manner:
General formula

Compound number R ₂ R ₃ R ₄
(25) CH = CH ₂ (CH ₂ ) ₃ cyclopropyl (26) CH = CH ₂ (CH ₂ ) ₃ isopropyl (27) CH ₃ (CH ₂ ) ₄ isopropyl (28) CH ₃ (CH ₂ ) ₄ cyclopropyl The product was identified by ¹ H-NMR.
Example V
Preparation of vitamin D compound (13) from ketone (24) Reaction formula:

(A). The free hydroxy group was protected by reaction with trimethylsilyl triflate (TBS-triflate) in methylene chloride as solvent in the presence of triethylamine at a temperature of -78 ° C --0 ° C. The yield of compound (29) was 80%.
(B). Enolization was performed in a corresponding manner as described in Example I (c) to yield compound (30) in 71% yield.
(C). The conjugation reaction with enine (6) was carried out in a corresponding manner as described in Example I (e) to give compound (31) in 94% yield.
(D). The final reaction step is performed in a corresponding manner as described in Example I (f), after which the protecting groups are removed using tetrabutylammonium fluoride as described in Example II (Desilylated). The final vitamin D compound (13) was obtained in 65% yield. The product was identical to the product obtained according to Example II.

The following vitamin D compounds were produced in a corresponding manner:
General formula

Compound number R ₂ R ₃ R ₄
(32) CH = CH ₂ (CH ₂ ) ₃ cyclopropyl (33) CH = CH ₂ (CH ₂ ) ₃ isopropyl (17) CH ₃ (CH ₂ ) ₄ isopropyl (16) CH ₃ (CH ₂ ) ₄ cyclopropyl The product was identified by ¹ H-NMR. The last two vitamin D compounds were identical to the corresponding vitamin D compounds prepared according to Example II.
Example VI
Preparation of 19-nor-vitamin D compound (35) from ketone (29) Reaction formula:

A solution of 1.14 g (2 mmol) of phosphine oxide (34) in 15 ml of dry THF was cooled to -78 ° C. N-Butyllithium (BuLi) in 2.5 M solution in hexane was added dropwise until the red color persisted. Next, 0.8 ml of a 2.5 M solution of BuLi was added. Stirring was continued for 15 minutes, after which 0.73 g (1.8 mmol) in 5 ml of THF was added dropwise. After stirring for a further hour, the reaction mixture was allowed to reach 0 ° C. and was then stopped by the addition of 50 ml of a saturated NH ₄ Cl solution. Extraction and flash chromatography (2% EtOAc in hexane) provided the protected diene compound. Desilylation by reaction with 10 equivalents of tetrabutylammonium fluoride (TBAF, 3 equivalents) in THF (10 ml) over 48 hours gave compound (35), which was flash chromatographed using EtOAc as eluent. And then recrystallized from MeOH / EtOAc. The overall yield was 53%.
Identification by ¹ H-NMR.

The following vitamin D compounds were produced in a corresponding manner:
General formula

Compound No. R ₂ R ₃ R ₄ AB
(36) CH = CH ₂ (CH ₂ ) ₃ isopropyl H H
(37) CH = CH ₂ (CH ₂ ) ₃ cyclopropyl H H
(38) CH ₂ OH (CH ₂ ) ₃ isopropyl H H
The product was identified by ¹ H-NMR.
Example VII
Affinity for Intracellular Vitamin D Receptor Vitamin D compounds according to the invention were dissolved in ethanol at concentrations ranging from 10 ^-13 to 10 ^-7 M. The affinity for calf thymus intracellular vitamin D receptor (VDR) was determined by a biological assay. In this assay, ³ H-1α, which is specifically bound to the VDR, 25-dihydroxy-cholecalciferol ^{(3 H-1α, 25-} DHCC) is replaced by the test compound. In particular, test compounds 11 , 13 and 14 had a high VDR-affinity. High VDR-affinity indicates a biologically active substance.
Example VIII
Affinity for Vitamin D Binding Protein Vitamin D binding protein (DBP) is a specific carrier for vitamin D and its metabolites in the blood. The biological activity of vitamin D compounds is dependent on their binding to DBP, since strong binding to DBP reduces intracellular accessibility to VDR. Weak binders are rapidly metabolized, which is desirable in topical applications.

In this assay, ³ H-1α and DBP, 25-DHCC and l [alpha], were cultured with several vitamin D compounds according with the present invention or 25-DHCC. For this purpose, the vitamin compounds were dissolved in ethanol at a concentration in the range of 10 ^{−11 to} 2.5 × 10 ⁻⁶ M. Bound / unbound ^{3 H-1α,} and then calculate the percentage of 25-DHCC. DBP was purified from whole human serum. The results are shown in the attached FIGS. 1 and 2. Figures 1 and 2 show the binding of vitamin D compounds to human vitamin D binding protein.
^[3 H] ^{_{1α, 25 (OH) 2 D}} 3 = 3 H-1α, 25-DHCC; In both figures, ● = 1α, 25-DHCC ( known compound); in FIG. 1, ▲ = compound 13 and ▽ = Compound 14 ; In FIG. 2, △ = Compound 11 .
Compounds 14 and 11 bind rather weakly to DBP as compared to the known 1α, 25-DHCC. Compound 13 is a very weak binder.
Example IX
Cell Differentiation Vitamin D compounds according to the invention were dissolved in ethanol at concentrations ranging from 10 ^-12 to 10 ^-6 M and tested for their ability to induce cell differentiation in the HL-60 assay. In this assay, morphological and biochemical tests of human leukocyte line HL-60 were performed to determine whether cell differentiation had occurred.

  Differentiation is expressed as the maturation factor nitroblue tetrazolium (NBT) meiotic non-specific esterase and is expressed as the percentage of mature cells beyond the myeloid stage that becomes visible after staining with May-Grunwald gemsa. After culturing with the known 1α, 25-DHCC or with the vitamin D compounds according to the invention, the percentage of cells containing black formazan deposits was determined. An increase in the percentage of NBT meiotic cells indicates an increase in cell differentiation.

By counting cell numbers and by the trypan blue exclusion method, the growth and viability of the cell culture is established. Cell viability and growth in HL-60 cultures were good in all conditions tested. 1α, 25-DHCC (known), Compound 12 , Compound 11 , Compound 13 and Compound 14 all induced differentiation and maturation of HL-60 cells. In cytological tests (non-specific esterases and May-Grunwald gemsa), especially compounds 13 and 11 were good differentiation agents. Optimal effects were seen at concentrations ranging from ^10-8 to ^10-7M .

The ability of compounds 13 and 14 to induce NBT-reduction was about 10-fold stronger than that of the known 1α, 25-DHCC. Compounds 11 and 12 were about 5 times more effective than 1α, 25-DHCC in inducing NBT-reduction (FIGS. 3 and 4).

  The above means that the tested novel vitamin D compounds of the present invention show higher cell differentiation activity than the known 1α, 25-DHCC.

Figures 3 and 4 (attached) show the differentiating effects of the tested vitamin D compounds on human leukocytes at the HL-60 line. In both drawings,
In FIG. 3, ＝= compound 13 and ▽ = compound 14 ; in FIG. 4, △ = compound 11 and ▽ = compound 12 .
Example X
The most well-known effect of 1α, 25-DHCC is its effect on calcium metabolism, ie the calcium-tropic effect. Calcium prone target organs are the intestine, bone and kidney.

Vitamin D compounds according to the invention were dissolved in ethanol and tested in the so-called Caco-2 assay for intestinal calcium (Ca) transport. In this assay, vitamin D-induced influx of ⁴⁵ Ca ²⁺ was measured in the monolayer of the intestinal cancer cell line Caco-2. This influx is corrected for concentration-induced Ca2 ⁺ influx, which is a measure for Ca transport across the intestinal wall. It is known that Caco-2 cells have a vitamin D receptor.

  Increasing intestinal calcium transport can be the first step in raising blood calcium levels (and indeed hypercalcemia).

Table A below shows the effect of the vitamin D compounds of the invention on Ca ²⁺ in intestinal Caco-2 cell cultures as compared to known 1α, 25-DHCC. The values in the table represent the relative increase in Ca ²⁺ influx (value for 1α, 25-DHCC is set at 100).

The results in Table A show that Compound 11 , Compound 13 and Compound 14 are stimulants of intestinal calcium absorption weaker than 1α, 25-DHCC.

Table A
Experiment 1:
Compound 10 ^-9 M
1α, 25-DHCC 100
Compound 11 79
Compound 12 99
Experiment 2:
Compound 10 ^-9 M
1α, 25-DHCC 100
Compound 13 65
Compound 14 78
Example XI
Calcium-tropic effects Along with intestines and bones, the kidney is also a major target organ of 1α, 25-DHCC. The kidney plays a very important role in calcium homeostasis because about 98% of calcium must be reabsorbed into the kidney to prevent calcium loss and hypercalcemia .

Vitamin D compounds according to the invention were dissolved in ethanol and tested in the rabbit kidney cell assay. In this assay, ⁴⁵ Ca ²⁺ reabsorption was measured in rabbit kidney cell monolayers. Cells were isolated by immunodissection of connecting tubes using monoclonal antibodies. Table B below shows the effect of the vitamin D compounds of the present invention on Ca ²⁺ -resorption in rabbit kidney cell cultures compared to the known 1α, 25-DHCC. Values in the table are ^{Ca 2+} - represents an increase in resorption in n mol / cm ² / hour.

The results in Table B show that Compound 13 and Compound 11 are weaker stimulators of renal Ca-reabsorption than Compound 12 and 1α, 25-DHCC itself.

Table B
Compound 10 ^-9 M
1α, 25-DHCC 13.4
Compound 11 2.4
Compound 12 12.8
Compound 13-3.7
Example XII
Cell differentiation to calcium趨性effective example known l [alpha], a drawback one that calcium趨性effect of such a 25-DHCC highly active vitamin D compounds, which harmful hypercalciuria, hypercalcemia and May cause urolithiasis. Therefore, it would be highly advantageous to develop compounds with high selectivity for biological activity. In other words, a compound in which the ratio between the induction of cell differentiation and a calcium trending effect such as, for example, stimulation of intestinal Ca transport is altered as compared to 1α, 25-DHCC.

The ratio between the differentiation-inducing capacity and the stimulation of intestinal Ca transport is the concentration at which 50% NBT reduction in HL-60 is obtained and at which half-maximal increase in intestinal Ca ²⁺ transport is reached. Defined as a fraction between. The lower the ratio, the higher the relative cell differentiation capacity.

  The results are shown in Table C.

Table C
Compound Fraction Ratio for DHCC 1α, 25-DHCC 107 1.00
Compound 12 29.7 0.28
Compound 11 14.4 0.13
Compound 14 2.6 0.02
Compound 13 2.5 0.02
Table C shows that the novel vitamin D compounds of the present invention that have been tested have better relative cell differentiation properties than the known 1α, 25-DHCC. Compounds 14 and 13 have a selective activity that is more than 50 times greater than 1α, 25-DHCC. Compound 11 has an 8 times better ratio. Compound 12 has a 4 times better ratio. This makes the novel vitamin D compounds of the present invention very suitable for applications where cell differentiation is desired (eg, in a hyperproliferative state).
The main features and aspects of the present invention are as follows.
1. General formula

(In the formula,
R ₁ is a hydrogen atom or a hydroxy group,
R ₂ is a (C ₁ -C ₃ ) alkyl group, a hydroxy (C ₁ -C ₃ ) alkyl group, a (C ₁ -C ₂ ) alkoxymethyl group or a (C ₂ -C ₃ ) alkenyl or alkynyl group;
R ₃ if and has at least 3 atoms with an epoxy, one or more of which do also be straight-chain or min substituted with a substituent selected from the full Oro and hydroxy in the main chain A branched, saturated or unsaturated aliphatic 3- to 5-membered hydrocarbon or oxahydrocarbon divalent group;
R ₄ is a sec- or tert- (C ₃ -C ₆ ) alkyl group or a (C ₃ -C ₆ ) cycloalkyl group, and A and B are each independently a hydrogen atom or a methyl group, or A and B together form a methylene group.
Vitamin D compounds.
2. R ₁ is a hydroxy group,
R ₂ has the meaning of 1 above,
R ₃ has the formula _{-O-CH 2 - (CH 2} ) n -, - CH 2 -CH 2 - (CH 2) n-,
-CH = CH- _(CH 2) n- or _{-CH 2 -CH 2 -CH (CH 3} ) -
Is a divalent group of
Where n is 1 or 2,
R ₄ is an isopropyl, cyclopropyl or tert-butyl group, and A and B are hydrogen atoms or together form a methylene group;
The vitamin D compound of claim 1 having the general formula I shown in 1 above.
3. R ₁ , R ₃ , A and B have the meaning of the above 2,
R ₂ has the meaning of 1 above and R ₄ is an isopropyl group,
The vitamin D compound of claim 1 having the general formula I shown in 1 above.
4. General formula

(In the formula,
R ₂ , R ₃ , A and B have the meaning of the above 1;
R ₅ is a protected hydroxy group, and R ₆ is _(C 1 -C ₆₎ alkyl group]
An ester compound of the general formula
R ₄ M (X) p (III)
(In the formula,
R ₄ has the meaning of the above 1;
X is Cl, Br or I;
M is a metal selected from Li and Mg, and p is 0 or 1 depending on the valency of M]
Characterized by reacting with an organometallic compound of, followed by removing the protecting group,
The method for producing a vitamin D compound according to the above 1, wherein R ₁ is a hydroxy group.
5. General formula

(In the formula,
R ₂ and R ₃ have the meaning of the above 1;
R ₅ ′ is an optionally protected hydroxy group, and R ₆ has the meaning of 4 above.
An ester compound of the general formula
R ₄ M (X) p (III)
(In the formula,
The symbol has the meaning of the above 4)
With an organometallic compound of the formula

When R ₅ ′ is a protected hydroxy group, the protecting group is removed and then oxidized to give a hydrindane compound having the general formula

Of the corresponding hydrindan-4-one compound of formula V and then, optionally after protection of the hydroxy group,
(A) General formula

(In the formula,
R ₅ has the meaning of the above 4;
R ₁ ′ is a hydrogen atom or a protected hydroxy group, and A and B have the meaning of the above 1)
Using the Wittig reagent of formula (I) or (b) after enolization and derivatization of the enolic hydroxy group,

(In the formula,
R ₁ ′ and R ₅ have the above-mentioned meanings]
And then hydrogenation and isomerization to produce a compound of general formula I in which A and B together form a methylene group, followed by removal of the protecting group. The method for producing the vitamin D compound according to 1 above, which is characterized in that:
6. R ₂ , R ₃ , A and B have the meaning of 1 above, and R ₅ and R ₆ have the meaning of 4 above,
An ester compound of the general formula IX represented by the above 4.
7. General formula

(In the formula,
R ₂ and R ₃ have the meaning of the above 1;
R ₆ has the meaning of 4 above and R ₇ is a derivatized hydroxy group.
An ester compound of the general formula

(In the formula,
R ₅ has the above-mentioned meaning]
Wherein A and B together form a methylene group, characterized by reacting with an enyne compound of formula IX, followed by hydrogenation and isomerization, Method.
8. General formula

(In the formula,
R ₂ and R ₃ have the meaning of 1 above, and R ₆ has the meaning of 4 above.
Of the general formula

(In the formula,
R ₅ has the meaning of 4 above, and A and B have the meaning of 1 above.
A process for the preparation of a compound of general formula IX as defined in 6 above, characterized by reacting with a Wittig reagent of formula (I).
9. General formula

(In the formula,
R ₂ , A and B have the meaning of the above 1;
R ₅ has the meaning of 4 above, and
n has the meaning of the above 2)
Iodide of general formula
CH ₂ = CHCOOR ₆
(In the formula,
R ₆ has the meaning of the above 4.
Wherein R ₃ is (CH ₂ ) ₃ or (CH ₂ ) _{4. A} process for the preparation of a compound of general formula IX as defined in 6 above, wherein R ₃ is (CH ₂ ) ₃ or (CH ₂ ) ₄ .
10. The hydrindan-4-one compound of the general formula V represented by the above 5, wherein the symbol has the meaning of the above 1.
11. Wherein the hydrindane compound of the general formula IV as defined in 5 above, wherein R ₅ ′ is a hydroxy group, is oxidized, preferably using an oxidizing agent selected from chromium-containing oxidizing agents and ruthenium tetroxide. A method for producing a hydrindan-4-one compound of general formula V as defined in 10 above.
12. A hydrindane compound of the general formula IV represented by 5 above, wherein R ₅ ′ has the meaning of 5 above and the other symbols have the meaning of 1 above.
13. Reacting a compound of general formula II as defined in 5 above with a metal-organic compound of general formula III as defined in 5 above in an inert organic solvent, characterized in that Of a hydrindane compound of the general formula IV as described above.
14． An effective amount of at least one compound as defined in 1, 2 or 3 above as active ingredient, together with a pharmaceutically acceptable carrier and / or at least one pharmaceutically acceptable auxiliary substance. Pharmaceutical compositions.
15. The composition according to claim 14 in dosage unit form for oral, topical (skin) or parenteral administration, containing from about 0.1 μg to about 0.1 mg of active ingredient per dosage unit.
16. Composition for diagnostic purposes which comprises, as active ingredients, an effective amount of at least one compound as defined in 1, 2 or 3 above, together with a compatible non-toxic carrier and / or at least one auxiliary substance. object.
17. An effective amount of at least one compound as defined above, 1, 2 or 3, as active ingredient, together with a non-toxic cosmetically acceptable carrier and / or at least one auxiliary substance. Is a cosmetic composition selected from the group consisting of creams, lipid creams, lotions, ointments, liposomes and gels.
18. Self-administration in a warm-blooded animal survivor, comprising administering to said warm-blooded animal survivor an effective amount for the intended purpose or treating said survivor with said composition. Treatment and prevention of numerous disease conditions including immune diseases, acne, alopecia, skin aging, imbalance of the immune system, inflammatory diseases such as rheumatoid arthritis and asthma, and diseases associated with abnormal cell differentiation and / or proliferation. Method.
19. 18. The method of claim 18 for the treatment of psoriasis and other hyperproliferative skin diseases.
20. Solid, skin or blood cancers, especially breast cancers, melanomas in warm-blooded animal survivors, comprising administering to the warm-blooded animal survivors the composition of 14 or 15 in an amount effective for this purpose. , Squamous cell carcinoma or leukemia.
21. A method of treating and preventing a number of skin conditions in a warm-blooded animal survivor, comprising administering to the warm-blooded animal survivor an effective amount of the cosmetic composition of claim 17.
22. 22. The method according to 21 above for the treatment and prevention of inadequate skin firmness or texture, insufficient skin hydration, wrinkles and / or insufficient sebum secretion.

FIG. 1 is a graph showing the binding of vitamin D compounds to human vitamin D-binding protein. FIG. 2 is a graph showing the binding of vitamin D compounds to human vitamin D-binding protein. FIG. 3 is a graph showing the discriminating effect of the tested vitamin D compounds on human leukocytes on the HL-60 line. FIG. 4 is a graph showing the discrimination effect of the tested vitamin D compounds on human leukocytes on the HL-60 line.

Claims (4)

General formula

(In the formula,
R ₂ represents a (C ₁ -C ₃ ) alkyl group, a hydroxy (C ₁ -C ₃ ) alkyl group, (C _1-
C ₂₎ alkoxymethyl group or _(C 2 -C ₃₎ alkenyl or alkynyl group,
R ₃ if and has at least 3 atoms with an epoxy, one or more of which do also be straight-chain or min substituted with a substituent selected from the full Oro and hydroxy in the main chain A branched, saturated or unsaturated aliphatic 3- to 5-membered hydrocarbon or oxahydrocarbon divalent group, and R ₄ is a sec- or tert- (C ₃ -C ₆ ) alkyl group or _(C 3 _-C 6)
A cycloalkyl group]
Hydrindane-4-one compound of formula (I). General formula

(In the formula,
R ₂ , R ₃ and R ₄ have the meaning of claim 1, and R ₅ ′ is a hydroxy group.
The hydrindan-4-compound of the general formula (V) according to claim 1, characterized in that the hydrindane compound having the formula (I) is oxidized preferably using an oxidizing agent selected from chromium-containing oxidizing agents and ruthenium tetroxide. A method for producing an on-compound. General formula

(In the formula,
R ₂ , R ₃ and R ₄ have the meaning of claim 1 and R ₅ ′ is an optionally protected hydroxy group.
Hydrindane compounds. General formula

(In the formula,
R ₂ and R ₃ have the meaning of claim 1,
R ₅ ′ is an optionally protected hydroxy group, and R ₆ is a (C ₁ -C ₆ ) alkyl group.
In an inert organic solvent, an ester compound of the general formula
R ₄ M (X) p (III)
(In the formula,
R ₄ has the meaning of claim 1,
X is Cl, Br or I;
M is a metal selected from Li and Mg, and p is 0 or 1 depending on the valency of M]
4. The method for producing a hydrindane compound of the general formula (IV) according to claim 3, wherein the method is reacted with an organometallic compound of the formula (4).

Images