VITAMIN D ANALOGS AND METHODS OF PREPARING THESE COMPOUNDS

Abstract: The invention relates to vitamin D analogs of general formula (I) wherein: R1 is a hydrogen atom or a substituent selected from the group consisting of hydroxy, CH2OH, CH2CH2OH, CH2CH2CH2OH, OCH3, OCH2OH, OCH2CH2OH and OCH2CH2CH2OH; R2 is a hydrogen atom or a substituent selected from the group consisting of OCH3, OCH2OH, CHm(CH2OH)n(CH2CH2OH)p, (CH2)qOH and O(CH2)rOH; wherein: m is 0 or 1, p, q and n are 0-3, r is 1-3 and m+n+p=3; with the proviso that R1 and/or R2 contain at least one OH group; R3 is a straight or branched, saturated or unsaturated aliphatic hydrocarbon of 6-13 C atoms which may be substituted with one or more substituents from the group hydroxy or fluoro. The invention further relates to methods of preparing these compounds, a pharmaceutical composition containing these compounds and to their use in pharmacotherapy and cosmetics.

Description: Vitamin D analogs and methods of preparing these compounds. The invention relates to new vitamin D analogs, to methods of preparing these compounds and to their use in pharmacotherapy and cosmetics. It is generally known, that vitamin-D compounds or vitamin-D related compounds ("vitamm-D analogs") have a strong biological activity and may be used m all those cases m which problems with the calcium and bone metabolism play a part. A few years ago it was found that various active vitamin-D compounds and analogs also have other pharmacotherapeutic activities and may be used successfully, for example, for the treatment of certain skm and bone diseases, for cosmetic applications and for treating diseases which are related to cell differentiation, cell proliferation or imbalance in the immune system, including diabetes mellitus, hypertension and inflammatory diseases such as rheumatoid arthritis and asthma. In addition, these compounds may be used in various veterinary applications, and for diagnostic purposes. Further it is known from J. Med. Chem. 23 (1995) , 4529-4537 that 1 α, 25-dihydroxy vitamin D„ analogs of the formula wherem Sj is OH, HOCH 2 - or H0CH 2 CH ^ have very low affinities, i.e. no more than 10 the affinity of calcitriol, for the calf thymus vitamin D receptor, and considerable antiproliferative activities m murine keratinocytes. Surprisingly it has now been found that a group of new vitamin D analogs has an activity of up to 6 times that of calcitriol, i.e. is much more active than the above mentioned known analogs, in the same test (affinity for the calf thymus vitamin D receptor) . The present invention relates to compounds of the general formula wherein R x is a hydrogen atom or a substituent selected from the group consisting of CH 2 OH, CH 2 CH 2 OH, CH 2 CH 2 CH 2 OH, OCH 3 , OCH 2 OH, OCH 2 CH 2 OH and OCH 2 CH 2 CH 2 OH; R 2 is a hydrogen atom or a substituent selected from the group consisting of 0CH 3 , OCH 2 OH, CH- (CH 2 OH) π (CH 2 CH 2 OH) p , (CH 2 ) σ OH and 0(CH 2 ) r OH; wherein m is 0 or 1, p, q and n are 0-3, r is 1-3 and m+n+p=3; with the proviso that R λ and/or R 2 contain at least one OH group; R 3 is a straight or branched, saturated or unsaturated aliphatic hydrocarbon of 6-13 C atoms which may be substituted with one or more substituents from the group hydroxy or fluoro. The above new vitamin D analogs of the invention, presented by the general formula I, are valuable substances The biological results, as illustrated in the Examples, indicate that these compounds are promising as biologically active substances and may be used m all above-mentioned pharmacotherapeutic indications, more in particular for the treatment of osteoporosis, renal osteodystrophy, osteomalacia, skm disorders such as psoriasis (and other hyperproliterative skm diseases) , eczema and dermatitis, myopathy, leukaemia, breast and cglon cancer, osteosarcomas, squamous cell carcinomas, melanoma, certain immunological disorders, and transplant rejections. Furthermore, the new vitamin D analogs of the invention may be used for wound healing and may be incorporated in cosmetic compositions, such as creams, lotions, ointments and the like, m order to preserve, condition and/or protect the skm and to improve various skm conditions, such as wrinkles, dry skm, skm slackness and insufficient sebum secretion. The new vitamin D analogs may also be used for diagnostic purposes. Preferred compounds of the general formula (I) are the compounds wherem R 3 is the group - (CH 2 ) 3 -C(CH 3 ) 2 OH Especially preferred are the compounds of formula (I) wherem R, is the group - (CH 2 ) 3 -C(CH 3 ) 2 0H and wherein: a) Rj . is a hydrogen atom and R 2 represents CH 2 CH 2 OH, CH 2 CH 2 CH 2 OH, CH(CH 2 CH 2 OH) 2 , C(CH 2 CH 2 OH) 3 or C(CH ? CH 2 0H) 2 CH 2 OH; or b) R x is OH, CH 2 OH or (CH 2 ) 2 OH and R ? represents hydrogen, OH, OCH 3 , or 0(CH 2 ) r 0H, wherem r has the value 1 to 3 Most particularly preferred are the compounds of formula (I) wherem R, is the group - (CH 2 ) 3 -C(CH 3 ) 2 OH and wherem R 3 is hydroxy, CH 2 OH and CH 2 CH 2 OH and R 2 represents hydrogen, CH 2 OH or CH 2 CH 2 OH. It is a special merit of the present mvention that the above new vitamin D analogs can easily be prepared from readily available starting materials. Consequently, the mvention also relates to a method of preparing a vitamin D analog of the general formula I, as defined above, which method is characterized in that a compound of the general formula wherem: R 3 ' is a straight or branched, saturated or unsaturated aliphatic hydrocarbon of 6-13 C-atoms which may be substituted with one or more substituents from the group protected hydroxy or fluoro, is reacted with an organometallic compound of the general formula R j ' and R 2 ' are the hydroxy protected analogs of Rj and R 2 defined above, X is Lι,MgCl, MgBr or Mgl, to yield a compound of the general formula followed by dehydration and deprotection of the hydroxy group (s) The compounds having formula I can also be obtained by reacting a compound of the general formula wherem R 3 ' has the meaning given above, and R 4 is a protected hydroxy group, with a compound of the general formula wherem X 1 is halogen, preferably Br, and R, ' and R 2 ' have the above given meanings, under the influence of an alkyl lithium compound, a zmc halogenide, preferably ZnCl 2 , and a transmetallation catalyst, followed by deprotection of the hydroxy group(s) . An example of a transmetallation catalyst is tris- (dibenzylidene-acetone) -dιpalladιum(O) The enolic hydroxy group of enolized compound IV is preferably derivatized by a reaction with N-aryltπflimide to produce a triflate. To improve the applicability of the new vitamin D analogs of the invention for the above-described pharmacotherapeutic indications, the compounds are usually processed to pharmaceutical compositions, comprising an effective amount of said vitamin D analog as the active ingredient in addition to a pharmaceutically acceptable carrier and/or at least one pharmaceutically acceptable auxiliary substance Such a composition may be delivered in a dosage unit form for oral, topical (dermal) or parenteral administration, comprising approx. 0.1 μg to approx. 0.1 mg active ingredient per dosage unit. A composition for diagnostic purposes may comprise, m addition to the vitamin D analog of the present invention, a compatible, non-toxic carrier and/or at least one auxiliary substance. A cosmetical composition may comprise, in addition to an effective amount (in the range of approx. 0.1 μg to. approx. 0.1 mg per dosage unit in a dosage unit form) of the vitamin D analog of the present invention, a cosmetically acceptable, non-toxic carrier and/or at least one auxiliary substance. Finally the invention relates to a method for the treatment and prophylaxis of a number of disease states including autoiummune diseases (including diabetes mellitus) , acne, alopecia, skin aging (including photo-aging) , imbalance m the immune system, inflammatory diseases such as rheumatoid arthritis and asthma, as well as diseases related to abnormal cell differentiation and/or proliferation, in a warm-blooded living being, comprising administering to said being or treating said being with a pharmaceutical composition as defined above in a quantity effective for the intended purpose. Examples of such diseases are psoriasis and other hyperproliferative skm diseases. The present invention also relates to the use of the above pharmaceutical compositions for the treatment of solid, skm and blood cancers, in particular of blood cancers such as leukaemia, of breast and colon cancer, and of skin cancers such as melanoma and squamous cell carcinoma. The above-defined cosmetical compositions, m particular selected from the group consisting of creams, lotions, ointments, liposomes and gels, can be used for the treatment and prevention of a number of skm disorders, such as inadequate skin firmness or texture, insufficient skm hydration, wrinkles and insufficient sebum secretion. ' The mvention will now be described in greater detail with reference to the following specific Examples. The following abbreviations are used in the examples: THF = tetrahydrofuran TBDMS = tert. -butyl dimethyl silyl DMF = N,N-dimethylformamide Pd 2 dba 3 = tris- (dibenzylideneacetone)dipalladium (0) Examples The TBDMS protected compound (Ilia) wherein X' = Br is dissolved in dry THF, cooled to -78°C and treated with 2 eq. of tert. C,H 9 Lι . After 30 minutes a solution of 1 eq. ZnCl 2 in dry THF is added and stirring is continued another 15 minutes. A solution of 1.1 eq. of a compound IV wherein R 4 is CF 3 S0 3 -, 5 mol % of the catalyst Pd 2 dba 3 and 10 mol % of As 3 P in dry DMF is introduced by means of double ended needle technique. The mixture is stirred at room temperature until the catalyst has completely dissolved. The progress of the reaction is monitored by thin-layer- chromatography. The reaction mixture is worked up by flash- chromatography. Complete deprotection of the hydroxy groups is carried out by treatment of the obtained product with an ion exchange resm, such as DOWEX-50 W, in methanol. The following compounds of formula (I) have been prepared m this manner: X H-NMR data (CDCl 3 ,δ) of compounds no.s 1-5: 400 mHz ^-NMR (CDCl 3 ,δ) : 0.76 (C 18 H 3 ; 3H; s) ; 1.00 (C 21 H 3 ; 3H, d) ; 1.22 (C 26 , 27 H 3 , 6H; s) ; 2.27 (CnH 2/ 2H, b) ; 2.58 (C 14 H; 1 H, b) ; 2.83 (Φ-CH 2 - C, 2x; 4H, t ) ; 3.85 (2x Φ-C-CH 2 -0, 4H, t) ; 5.64 (C 9 H; IH, b) ; 6.92 (A, IH, s) ; 6.93 (B; 2H, s) . 400 mHz X H-NMR (CDC1 3/ δ) : 0.73 C 18 H 3 ; 3H,s) ; 0.99 (C 21 H 3 , 3H, d) ; 1.19 (C 26,27 H 3 ; 6H,S) ; 2.23 (C 11 H 2 , 2H, b) ; 2.48 (C 14 H, IH, b) ; 5.58 (C 9 H, IH, b) ; 6.16 (B, 2H, s) ; 6.20 (A, IH, s) ; 8.53 (2xΦ-OH, 2H, s) . No. 3 400 mHz Η-NMR (CDC1 3 , δ) : 0.77 (C 18 H 3 ; 3H, s) ; 1.00 (C 21 H 3/ 3H, d) ; 1.21 (C 26>27 H 3 ; 6H, s) ; 2.58 (C 14 H, IH, b) ; 4.65 (Φ-CH 2 -0, 2H, s) ; 5.63 (CyH; IH, b) ; 7.18 (Φ-H, 2H, t) ; 7.26 (Φ-H, 2H, b) . 400 mHz Η-NMR (CDCl 3 , δ) : 0.76 (C le H 3 , 3H, s) ; 1.00 (C 21 H 3 ; 3H, d); 1.22 (C 26-27 H 3 , 6H, s) ; 2.60 (C 14 H, IH, b) ; 2.84 (Φ-CH 2 -C, 2H, t) ; 3.85 (Φ-C-CH 2 -0, 2H, t) ; 5.62 (C 9 H, 1 H, b) ; 7.14 (Φ-H, 4H, b) . 400 mHz Η-NMR (CDC1 3 , δ) : 0.76 (C 1B H 3/ 3H, s) ; 1.00 (C 21 H 3 , 3H, d) ; 1.22 (C 26<27 H 3/ 6H, s) ; 2.59 (C 14 H, IH, b) ; 4.64 (2x Φ-CH 2 -0, 4H, S) ; 5.65 (C 9 H; IH, b) ; 7.11 (2x B; 2H, S ) ; 7.18 (A, IH; s) . Affinity towards the intracellular vitamin D receptor The aromatic vitamin D analogs were dissolved in ethanol in concentrations ranging from 10 14 to IO "6 M. The affinity towards the calf thymus intracellular vitamin D receptor (VDR) was determined in a biological in vitro assay and compared to that of calcitriol. In this assay, labelled Η-calcitπol, which is specifically bound to the VDR, is replaced by the aromatic vitamin D analog. Compounds 5 and 3 have even higher affinities (resp. 6 and 3 times) than calcitriol itself towards the highly selective VDR. Also analogs 4 and 1 demonstrate high VDR affinities. A high VDR affinity is indicative for biologically active substances. The compounds with the highest affinities were therefore further tested m. vitro. Affinity to human blood vitamin D binding protein Vitamin D binding protein (DBP) is the specific carrier for vitamin D and its metabolites in blood. The biological activity of vitamin D analogs depends on their binding to DBP. Strong binders will have reduced access to the VDR. Weak binders are rapidly metabolized, which is a favourable aspect m topical application. DBP is purified from total human serum In the assay, DBP is incubated with 3 H-calcιtrιol and calcitriol or one of the aromatic vitamin D analogs. To this purpose, the vitamin D analogs are dissolved in ethanol m concentrations ranging from 10 n to 2.5 X 10 6 . The percentage bound/unbound Η-calcitriol is then calculated. Compounds 5, 4 and 3 (m order of binding) are relatively good binders, binding a factor 2-10 less than calcitriol itself. These binding capacities show that the compounds might have systemic actions, i.e. they might be active when administrated orally or given by injection.

Claims: Claims 1. A vitamin D analog of the general formula wherem: Rj is a hydrogen atom or a substituent selected from the group consisting of CH 2 OH, CH 2 CH 2 OH, CH 2 CH 2 CH 2 OH, 0CH 3 , OCH 2 OH, OCH 2 CH 2 OH and OCH 2 CH 2 CH 2 OH; R 2 is a hydrogen atom or a substituent selected from the group consisting of OCH 3 , 0CH 2 0H, CH m (CH 2 OH) n (CH 2 CH 2 OH) p , (CH 2 ) q OH and 0(CH 2 ) r OH; wherem: m is 0 or 1, p, q and n are 0-3, r is 1-3 and m+n+p=3,- with the proviso that R 3 and/or R 2 contain at least one OH group; R 3 is a straight or branched, saturated or unsaturated aliphatic hydrocarbon of 6-13 C atoms which may be substituted with one or more substituents from the group hydroxy or fluoro. 2. A compound as claimed m Claim 1, having the general formula I wherein R 3 is the group - (CH 2 ) 3 -C(CH 3 ) 2 OH. 3. A compound as claimed in Claim 2, having the general formula I wherein: (a) Ri is a hydrogen atom and R 2 represents CH 2 CH 2 OH, CH 2 CH 2 CH 2 OH, CH(CH 2 CH 2 OH) 2 , C(CH 2 CH 2 OH) 3 or C(CH 2 CH 2 OH) 2 CH 2 OH; or (b) Rj is OH, CH 2 OH or (CH 2 ) 2 OH and R 2 represents hydrogen, OH, OCH 3 , or 0(CH 2 ) r OH, wherem r has the value 1 to 3. 4. A compound as claimed in Claim 2, having the general formula I wherem: R α is hydroxy, CH 2 0H and CH 2 CH 2 0H and R 2 represents hydrogen, CH 2 OH or CH 2 CH 2 0H. 5. A method of preparing a vitamin D analog as claimed in Claim 1, characterized in that a compound of the general formula wherem: R 3 ' is a straight or branched, saturated or unsaturated aliphatic hydrocarbon of 6-13 C atoms which may be substituted with one or more substituents from the group protected hydroxy or fluoro, is reacted with a compound of the general formula wherem: R j ' and R 2 ' are the hydroxy protected groups Rj and R 2 as defined in Claim 1, X is Li, MgCl, MgBr or Mgl , to yield a compound of the general formula followed by dehydration and deprotection of the hydroxy groups. 6. A method of preparing a vitamin D analog as claimed in Claim 1, characterized in that a compound of the general formula wherem: R 3 ' has the meaning given in claim 5; and R 4 is a protected hydroxy group, is reacted with a compound of the general formula wherein: X' is halogen, preferably Br; R 3 ' and R 2 ' are the hydroxy protected groups R 3 and R 2 as defined in Claim 1; under the influence of an alkyl lithium compound, a zmc halogenide, preferably ZnCl 2 , and a transmetallation catalyst, followed by deprotection of the hydroxy groups. 7. A pharmaceutical composition comprising, m addition to a pharmaceutically acceptable carrier and/or at least one pharmaceutically acceptable auxiliary substance, as the active ingredient at least one compound as defined in Claim 1 to 4 in an effective amount. 8. A method of preparing pharmaceutical compositions as claimed in claim 7, characterized in that a compound as claimed m claim 1-4 is brought into a form suitable for administration. 9. A method for the treatment and/or prevention of a number of skm disorders or of a number of diseases, including bone diseases such as osteoporosis, renal osteodystrophy and osteomalacia, autoimmune diseases, acne, alopecia, skm aging, imbalance in the immune system, inflammatory diseases such as rheumatoid arthritis and asthma, as well as diseases related to abnormal cell differentiation and/or proliferation, as well as solid, skin or blood cancers, in a warm-blooded living being, comprising administering to said being or treating said being with a composition as claimed m Claim 7 in a quantity effective for the intended purpose .

Inventors: HALKES SEBASTIAN, KANZLER SILVIA, REISCHL WOLFGANG, VAN DE VELDE JAN PAUL, HALKES, SEBASTIAN, KANZLER, SILVIA, REISCHL, WOLFGANG, VAN DE VELDE, JAN, PAUL
Applicants: DUPHAR INT RES, HALKES SEBASTIAN, KANZLER SILVIA, REISCHL WOLFGANG, VELDE JAN PAUL VAN DE, DUPHAR INTERNATIONAL RESEARCH B.V., HALKES, SEBASTIAN, KANZLER, SILVIA, REISCHL, WOLFGANG, VAN DE VELDE, JAN, PAUL
Application year: 1997
Main classification: C07C33
IPC codes: A61K31, A61P3, C07C29, C07C33, C07C37, C07C39