DK169945B1

DK169945B1 - 25-Hydroxy Vitamin D3 Derivatives, Process for Preparation thereof and Drug Containing These Compounds

Info

Publication number: DK169945B1
Application number: DK019789A
Authority: DK
Inventors: Bernard Michael Hennessy; Shian-Jan Shiuey; Gary Arthur Truitt; Milan Radoje Uskokovic; Enrico Giuseppe Baggiolini
Original assignee: Hoffmann La Roche
Priority date: 1988-01-20
Filing date: 1989-01-17
Publication date: 1995-04-10
Also published as: IL88989A0; ES2033467T3; AU2864489A; BG60530B2; FI890283A7; DK19789A; NO175429B; MC1998A1; NO175429C; JPH0764806B2; ZA8923B; JPH029861A; CA1337529C; FI90764B; IE890157L; PT89486A; IE60921B1; FI890283A0; PT89486B; KR960009119B1

Description

DK 169945 B1DK 169945 B1

Opfindelsen angår hidtil ukendte 25-hydroxy-vitamin-Dj-derivater, som er ejendommelige ved, at de har den almene formel IThe invention relates to novel 25-hydroxy-vitamin D 1 derivatives which are characterized in that they have the general formula I

rtTVw 5 hvor R er hydrogen eller hydroxy, og A er -C-C-, -CH=CH- med E- konfiguration eller -CH2-CH2-, disse forbindelser til anvendelse som terapeutisk aktive stoffer, især til behandling af hyperproliferative hudsygdomme, især psoriasis, eller til behandling af neoplastiske sygdomme, især leukæmi, en 10 fremgangsmåde til fremstilling af disse forbindelser, der er ejendommelig ved, at en tilsvarende forbindelse med formlen I, der i stedet for to eller tre hydroxygrupper indeholder to eller tre beskyttede hydroxygrupper med formlen -OSi(R^,R2,R3) 15 hvor R2 og R3 er C^.4-alkyl, og R2 er <^.4-alkyl, aryl eller aryl-C1.4-alkyl, omsættes med et middel, der er i stand til at fjerne beskyttelsesgrupperne, et lægemiddel, især til behandling af hyperproliferative hudsygdomme, især psoriasis, eller til behandling af neoplastiske 20 sygdomme, især leukæmi, omfattende en effektiv mængde af en forbindelse med formlen I og et farmaceutisk effektivt bæremateriale, især til oral eller topisk administration, samt anvendelse af en forbindelse med formlen I til fremstilling af et lægemiddel til behandling af hyperproliferative hudsygdomme, især psoriasis, eller til behand-25 ling af neoplastiske sygdomme, især leukæmi.where R is hydrogen or hydroxy and A is -CC-, -CH = CH- with E-configuration or -CH2-CH2-, these compounds for use as therapeutically active substances, especially for the treatment of hyperproliferative skin diseases, especially psoriasis , or for the treatment of neoplastic diseases, especially leukemia, a process for the preparation of these compounds, characterized in that a corresponding compound of formula I containing, instead of two or three hydroxy groups, contains two or three protected hydroxy groups of formula - OSi (R 1, R 2, R 3) wherein R 2 and R 3 are C 1-4 alkyl and R 2 is C 1-4 alkyl, aryl or aryl C 1-4 alkyl are reacted with an agent which is in capable of removing the protecting groups, a drug, in particular for the treatment of hyperproliferative skin diseases, especially psoriasis, or for the treatment of neoplastic diseases, especially leukemia, comprising an effective amount of a compound of formula I and a pharmaceutically effective carrier, particularly for oral ortopical administration, and the use of a compound of formula I for the manufacture of a medicament for the treatment of hyperproliferative skin diseases, especially psoriasis, or for the treatment of neoplastic diseases, especially leukemia.

DK 169945 B1 2DK 169945 B1 2

Det har overraskende vist sig, at disse forbindelser forårsager mindre uønsket kalcifikation af blødt væv ved subkutan administration end la,25-dihydroxy-vitamin-D3 ved behandlingen af ovennævnte sygdomme .Surprisingly, these compounds have been found to cause less undesirable soft tissue calcification by subcutaneous administration than 1α, 25-dihydroxy vitamin D3 in the treatment of the above diseases.

5 Eksempler på de nedenfor nævnte C-|__4-alkyl grupper er methyl, ethyl, „ propyl, isopropyl, butyl og t-butyl. Eksempler på aryl-C-L.^alkylgrupper er benzyl, phenethyl og phenylpropyl. Eksempler på arylgrupper er phenyl og p-tolyl. Halogen betegner brom, chlor, fluor eller iod.Examples of the C 1-4 alkyl groups mentioned below are methyl, ethyl, "propyl, isopropyl, butyl and t-butyl. Examples of aryl C 1 -C 6 alkyl groups are benzyl, phenethyl and phenylpropyl. Examples of aryl groups are phenyl and p-tolyl. Halogen represents bromine, chlorine, fluorine or iodine.

Forbindelser med formlen I ifølge opfindelsen er de nedenfor defi-10 nerede forbindelser A til F: A: 1,25 -dihydroxy-16 -dehydrocholecalcif erol ,-B: 25-hydroxy-16-dehydrocholecalciferol; C: 1,25-dihydroxy-16,23E-bisdehydrocholecalciferol; D: 25-hydroxy-16,23E-bisdehydrocholecalciferol; 15 E: l,25-dihydroxy-16-dehydro-23-didehydrocholecalciferol; og F: 25-hydroxy-16-dehydro-23-didehydrocholecalciferol; blandt disse foretrækkes de 1,25-dihydroxylerede forbindelser A, C og E.Compounds of formula I according to the invention are the compounds defined below A to F: A: 1,25-dihydroxy-16-dehydrocholecalciferol, -B: 25-hydroxy-16-dehydrocholecalciferol; C: 1,25-dihydroxy-16,23E-bisdehydrocholecalciferol; D: 25-hydroxy-16,23E-bisdehydrocholecalciferol; E: 1,25-dihydroxy-16-dehydro-23-didehydrocholecalciferol; and F: 25-hydroxy-16-dehydro-23-didehydrocholecalciferol; of these, 1,25-dihydroxylated compounds A, C and E. are preferred.

Forbindelserne med formlerne la og Ib (omfattet af formel I) kan 20 fremstilles som beskrevet i skemaerne 1, 2 og 3 ved omsætning af en tilsvarende forbindelse med formlen I, der i stedet for de to eller tre hydroxygrupper indeholder to eller tre beskyttede hydroxygrupper med formlen -OSi (R^ R2, R3) * 25 hvor R1 og R3 er C1.4-alkyl, og R2 er C1.4-alkyl, aryl eller aryl-C1.4-alkyl, med et middel, der er i stand til at fjerne beskyttelsesgrupperne.The compounds of formulas Ia and Ib (comprised of Formula I) can be prepared as described in Schemes 1, 2 and 3 by reacting a corresponding compound of Formula I containing, instead of the two or three hydroxy groups, two or three protected hydroxy groups having the formula -OSi (R 1, R 2, R 3) * wherein R 1 and R 3 are C 1-4 alkyl and R 2 is C 1-4 alkyl, aryl or aryl C 1-4 alkyl, with an agent capable of to remove the protecting groups.

Skema ISchedule I

3 DK 169945 B1 ΛΚ I cs:—Sj o H n ( , v^a / a,3 DK 169945 B1 ΛΚ I cs: —Sj o H n (, v ^ a / a,

I CSi—Js. IIn CSi — Js. IN

JH γΐΤ7 i,JH γΐΤ7 i,

Ju JCJu JC

f fY ? i’ cYf fY? i 'cY

Rj-S.-o-'--\^S0_s;_.2 Bj-Si-O'-X/Rj-S.-o -'-- \ ^ S0_s; _. 2 Bj-Si-O'-X /

β, i Iβ, i

3 r3 *3 r3 *

Wi ivb rj^P^ rtTV- jrWi ivb rj ^ P ^ rtTV- jr

•UT CJ• UT CJ

"O" ^CH MO···^ h b hvor A har den ovenfor anførte betydning, og R^ og R3 hver for sig er c1-4-a3.ky1, og hvert R2 for sig er C1.4-alkyl, aryl eller aryl-C·^-alkyl"O" ^ CH MO ··· ^ hb where A has the meaning given above and R 1 and R 3 are each c1-4-a3.ky1 and each R2 is individually C1-4 alkyl, aryl or aryl-C ^ -alkyl ·

Skema 2 4 DK 169945 B1Scheme 2 4 DK 169945 B1

KOCOW

VV

% r+pV CpT^"% r + pV CpT

|H HO| H HO

O V! VIIO V! WE YOU

iin

O VIIIO VIII

Mb . ^j" CSI— =2 I Sj 0 Ila hvor R-j_/ R2 og R3 har de ovenfor anførte betydninger.Mb. CSI - = 2 In Sj0Ila where R-j_ / R2 and R3 have the meanings given above.

Skema 3 5 DK 169945 B1 0Τ» /ψί Γ „ I i \ I OSi R V^7 "sScheme 3 5 DK 169945 B1 0Τ »/ ψί Γ" I i \ I OSi R V ^ 7 "s

RiRi

IXIX

R, · "iR, · "i

Ri—SiC^^HRi-SiC ^^ H

t ΛΙt ΛΙ

Ri HO A" o osl—a2 TA nc o hvor R1# R2 og R3 har de ovenfor anførte betydninger, X er chlor, brom eller iod, og Ts er tosyl.R 1 H 2 O 2 - a 2 TA nc o wherein R 1 # R 2 and R 3 have the above meanings, X is chlorine, bromine or iodine and Ts is tosyl.

6 DK 169945 B16 DK 169945 B1

Mellemprodukterne med formlen II, herunder de mellemprodukter, der har formlerne Ila, Ilb og Ile, er hidtil ukendte.The intermediates of formula II, including the intermediates having formulas Ila, Ilb and Ile, are novel.

I skema 1 omdannes forbindelsen med formlen II til en forbindelse med formlen IVa eller IVb ved omsætning med den tilsvarende forbindelse 5 med formlen I^POFhj 1' .Cl R2-SiO-'%,Ss^R4 111 r3 hvor Ph er phenyl; R4 er H eller -OSi (R1(R2 R3) , og Rlf R2 og R3 har de ovenfor anførte betydninger.In Scheme 1, the compound of formula II is converted to a compound of formula IVa or IVb by reaction with the corresponding compound 5 of formula I ^ POFhj 1 '.Cl R2-SiO -'%, Ss ^ R4 111 r3 where Ph is phenyl; R4 is H or -OSi (R1 (R2 R3) and R1f R2 and R3 have the meanings given above).

10 Reaktionen udføres ved -60 til 90°C, fortrinsvis -75eC, i et polært, aprot, organisk opløsningsmiddel, fx tør ether eller fortrinsvis tør tetrahydrofuran (THF), i nærværelse af en stærk base såsom en alkyl-lithium, fx butyllithium.The reaction is carried out at -60 to 90 ° C, preferably -75 ° C, in a polar, aprotic organic solvent, e.g. dry ether or preferably dry tetrahydrofuran (THF), in the presence of a strong base such as an alkyl lithium, eg butyllithium.

Beskyttelsesgrupperne på en forbindelse med formlen IVa eller IVb 15 fjernes ved reaktion med et fluorsalt, fx tetrabutylammoniumfluorid, i et polært, organisk opløsningsmiddel, fx ether eller fortrinsvis THF, til opnåelse af en tilsvarende forbindelse med formlen la eller Ib.The protecting groups on a compound of formula IVa or IVb are removed by reaction with a fluorine salt, e.g., tetrabutylammonium fluoride, in a polar organic solvent, e.g., ether or preferably THF, to give a corresponding compound of formula Ia or Ib.

I skema 2 oxideres forbindelsen med formlen V til forbindelsen med 20 formlen VI ved behandling med et oxidationsmiddel, fx 2,2'-bipyridi-niumchlorchromat eller fortrinsvis pyridiniumchlorchramat, i et aprot, organisk opløsningsmiddel såsom methylenchlorid. iIn Scheme 2, the compound of formula V is oxidized to the compound of formula VI by treatment with an oxidizing agent, for example 2,2'-bipyridinium chlorochromate or preferably pyridinium chlorochramate, in an aprotic organic solvent such as methylene chloride. in

Forbindelsen med formlen VI omdannes til en forbindelse med formlen Ilb ved omsætning med fx en (trialkylsilyl) imidazol såsom (trimethyl-25 silyl) imidazol i et aprot organisk opløsningsmiddel såsom THF eller fortrinsvis methylenchlorid.The compound of formula VI is converted to a compound of formula IIb by reaction with, for example, a (trialkylsilyl) imidazole such as (trimethylsilyl) imidazole in an aprotic organic solvent such as THF or preferably methylene chloride.

7 DK 169945 B17 DK 169945 B1

Forbindelsen med formlen V også kan blive delvist hydrogeneret til forbindelsen med formlen VII ved omsætning med et reduktionsmiddel, fx lithiumaluminiumhydrid, fortrinsvis i nærværelse af et alkali-metalalkoxid, fx natriummethoxid, i et aprot organisk opløsnings-5 middel, fx ether eller fortrinsvis THF, ved tilbagesvalingstemperatur (ca. 68°C for THF), i ca. 10-20 timer.The compound of formula V may also be partially hydrogenated to the compound of formula VII by reaction with a reducing agent, eg lithium aluminum hydride, preferably in the presence of an alkali metal alkoxide, eg sodium methoxide, in an aprotic organic solvent, eg ether or preferably THF, at reflux temperature (about 68 ° C for THF), for approx. 10-20 hours.

Den resulterende forbindelse med formlen VII oxideres til forbindelsen med formlen VIII ved behandling med et oxidationsmiddel som beskrevet ovenfor for oxidationen af V til VI.The resulting compound of formula VII is oxidized to the compound of formula VIII by treatment with an oxidizing agent as described above for the oxidation of V to VI.

10 Forbindelsen med formlen VIII omdannes til en forbindelse med formlen Ila ved omsætning med en (trialkylsilyl)imidazol som beskrevet ovenfor for omdannelsen af VI til Ilb.The compound of formula VIII is converted to a compound of formula IIa by reaction with a (trialkylsilyl) imidazole as described above for the conversion of VI to IIb.

I skema 3 omsættes forbindelsen med formlen X med magnesium i ether, fortrinsvis THF, ved tilbagesvalingstemperatur. Den resulterende 15 Grignard-opløsning behandles med cuproiodid, og derefter tilsættes forbindelsen med formlen IX.In Scheme 3, the compound of formula X is reacted with magnesium in ether, preferably THF, at reflux temperature. The resulting Grignard solution is treated with cuprous iodide and then the compound of formula IX is added.

Den resulterende forbindelse med formlen XI omsættes med et fluoridsalt, fx tetrabutylammoniumfluorid, i ether eller fortrinsvis THF.The resulting compound of formula XI is reacted with a fluoride salt, e.g., tetrabutylammonium fluoride, in ether or preferably THF.

Den vundne forbindelse med formlen XII kan oxideres som beskrevet 20 ovenfor for oxidationen af V til VI.The compound of formula XII obtained can be oxidized as described above for the oxidation of V to VI.

Den resulterende forbindelse med formlen XIII omdannes til en forbindelse med formlen Ile ved omsætning med en (trialkylsilyl)imidazol som beskrevet ovenfor for omdannelsen af VI til Ilb.The resulting compound of formula XIII is converted to a compound of formula Ile by reaction with a (trialkylsilyl) imidazole as described above for the conversion of VI to IIb.

Til fremstilling af en forbindelse med formlen IX kan forbindelsen 25 med formlenFor the preparation of a compound of formula IX, compound 25 of formula

V^OHV ^ OH

8 DK 169945 B18 DK 169945 B1

XIVXIV

ho Hho H

(Tetrahedron 40, 1984, 2283) omsættes med et tosyleringsmiddel såsom et p-toluensulfonylhalogenid, fx chloridet, i en organisk base, fx 5 collidin eller fortrinsvis pyridin. Den resulterende forbindelse med formlen(Tetrahedron 40, 1984, 2283) is reacted with a tosylating agent such as a p-toluenesulfonyl halide, e.g., the chloride, in an organic base, e.g., collidine or preferably pyridine. The resulting compound of the formula

VoTs xvVoTs xv

1 H1 H

HOHAY

omdannes derefter til en forbindelse med formlen IX ved omsætning af 10 et trialkylsilylchlorid, fx trimethylsilylchlorid, i nærværelse af imidazol og i et aprot organisk opløsningsmiddel, fx THF eller methy-lenchlorid.is then converted to a compound of Formula IX by reaction of a trialkylsilyl chloride, e.g. trimethylsilyl chloride, in the presence of imidazole and in an aprotic organic solvent, e.g. THF or methylene chloride.

Til fremstilling af en forbindelse med formlen X kan en forbindelse med formlen 15 X-CH2CH2COCH3 XVi omdannes til en forbindelse med formlenTo prepare a compound of formula X, a compound of formula 15 X-CH 2 CH 2 COCH 3 XVi can be converted to a compound of formula

X-CH2CH2C(CH3)2-OH XVIIX-CH2CH2C (CH3) 2-OH XVII

hvor X har den ovenfor anførte betydning, ved omsætning med et me- £ thyl-Grignard-reagens såsom methylmagnesiumbromid i ether. Forbindel-20 sen med formlen XVII omdannes til en forbindelse med formlen X ved omsætning med et trialkylsilylchlorid som beskrevet ovenfor for omdannelsen af XV til IX.wherein X is as defined above by reaction with a methyl Grignard reagent such as methyl magnesium bromide in ether. The compound of formula XVII is converted to a compound of formula X by reaction with a trialkylsilyl chloride as described above for the conversion of XV to IX.

a 9 DK 169945 B1a 9 DK 169945 B1

Til fremstilling af forbindelsen med formlen V omsættes forbindelsen med formlen XV ovenfor med et cyaniddannende middel, fx natriumcyanid, i et aprot organisk opløsningsmiddel, fx dimethylsulfoxid (DMSO), ved en temperatur mellem 80 og 100eC i 1-5 timer til opnåelse 5 af en forbindelse med formlen v^cnTo prepare the compound of formula V, the compound of formula XV above is reacted with a cyanide-forming agent, e.g., sodium cyanide, in an aprotic organic solvent, e.g., dimethyl sulfoxide (DMSO), at a temperature between 80 and 100 ° C for 1-5 hours to give 5 compound of the formula v ^ cn

XVIIIXVIII

HO *HO *

Denne omdannes til forbindelsen med formlen i * Ύ choThis is converted into the compound of the formula in * Ύ cho

117 XIX117 XIX

HO HHO H

10 ved omsætning med et reduktionsmiddel, fx diisobutylaluminiumhydrid, efterfulgt af hydrolyse med fx en mineralsyre såsom saltsyre. Reduktionen udføres i et aprot organisk opløsningsmiddel, fx methylen-chlorid, ved ca. -10 til 10°C i ca. 20 til 90 minutter. Forbindelsen 15 med formlen XIX omdannes til forbindelsen med formlen s Λ tf 'V8' CXy * xx10 by reaction with a reducing agent, for example diisobutyl aluminum hydride, followed by hydrolysis with, for example, a mineral acid such as hydrochloric acid. The reduction is carried out in an aprotic organic solvent, e.g., methylene chloride, at approx. -10 to 10 ° C for approx. 20 to 90 minutes. The compound 15 of the formula XIX is converted to the compound of the formula such as 'V8' CXy * xx

HO HHO H

ved omsætning med en blanding af triphenylphosphin, carbontetrabromid og zinkstøv i et aprot organisk opløsningsmiddel, fx methylenchlorid, 20 i ca. 1 til 30 timer.by reaction with a mixture of triphenylphosphine, carbon tetrabromide and zinc dust in an aprotic organic solvent, for example methylene chloride, for about 20 hours. 1 to 30 hours.

10 DK 169945 B110 DK 169945 B1

Forbindelsen med formlen XX omdannes til forbindelsen med formlenThe compound of formula XX is converted to the compound of formula

H XXIH XXI

HO HHO H

ved omsætning med en stærk base, fx butyllithium, i et polært aprot 5 opløsningsmiddel, fx THF, ved ca. -80 til -70eC i ca. 1-3 timer. Forbindelsen med formlen XXI omdannes til forbindelsen med formlenby reaction with a strong base, e.g., butyllithium, in a polar aprot 5 solvent, e.g., THF, at ca. -80 to -70 ° C for approx. 1-3 hours. The compound of formula XXI is converted to the compound of formula

HH

XXIIXXII

MejSiOMejSiO

ved omsætning med (trimethylsilyl) imidazol i et aprot organisk op-10 løsningsmiddel, fx THF eller methylenchlorid. Denne forbindelse omdannes til forbindelsen med formlen s » Γ T^y I ch xxiiiby reaction with (trimethylsilyl) imidazole in an aprotic organic solvent, eg THF or methylene chloride. This compound is converted to the compound of formula s »Γ T ^ y I ch xxiii

Me3SiO HMe3SiO H

ved omsætning med en stærk base, fx butyllithium, og derefter med 15 acetone. Reaktionen udføres i et aprot organisk opløsningsmiddel, fx THF, ved ca. -80 til -60°C. Forbindelsen med formlen XXIII afbeskyt-tes til opnåelse af forbindelsen med formlen V (i skema 2) ved omsætning med et fluorsalt, fx tetrabutylammoniumfluorid, i et organisk opløsningsmiddel, fx ether eller THF.by reaction with a strong base, for example, butyllithium, and then with 15 acetone. The reaction is carried out in an aprotic organic solvent, e.g. THF, at ca. -80 to -60 ° C. The compound of formula XXIII is deprotected to give the compound of formula V (in Scheme 2) by reaction with a fluorine salt, eg tetrabutylammonium fluoride, in an organic solvent, eg ether or THF.

20 Forbindelsen med formlen I stimulerer differentiering og nedsætter proliferation af humane keratinocytter. De er som følge heraf nyttige som midler til behandling af hyperproliferative hudsygdomme såsom 11 DK 169945 B1 psoriasis, basalcelle-carcinomer, -lidelser eller -keratinisering og keratosis. Forbindelsen med formlen I er også nyttige som midler til behandling af neoplastiske sygdomme såsom leukæmi.The compound of formula I stimulates differentiation and slows down the proliferation of human keratinocytes. As a result, they are useful as agents for the treatment of hyperproliferative skin diseases such as psoriasis, basal cell carcinomas, disorders or keratinization and keratosis. The compound of formula I is also useful as agents for the treatment of neoplastic diseases such as leukemia.

Model for kalcifikation af blødt væv 5 Undersøgelsens formål var at vurdere kalcifikationen af bløde væv forårsaget af forbindelserne ifølge opfindelsen. På undersøgelsens første dag blev rotter mærket med en enkelt subkutan injektion af 40 μΟί 4^Ca. Forbindelserne blev derefter administreret enten subkutant eller topisk i fire på hinanden følgende dage. Rotterne blev 10 aflivet ved C02-inhalation 24 timer efter den sidste injektion.Model for Soft Tissue Calcification 5 The purpose of the study was to assess the soft tissue calcification caused by the compounds of the invention. On the first day of the study, rats were labeled with a single subcutaneous injection of 40 μΟί 4 ^ Ca. The compounds were then administered either subcutaneously or topically for four consecutive days. The rats were sacrificed by CO 2 inhalation 24 hours after the last injection.

Hjerterne og nyrerne blev udtaget, anbragt i scintillationstælleglas og opløst i 24 timer med 2,0 ml salpetersyre. En alikvot (0,2 ml) af det opløste materiale blev dernæst sat til 9,8 ml Aquasol og talt i en scintillationstæller.The hearts and kidneys were removed, placed in scintillation counters and dissolved for 24 hours with 2.0 ml of nitric acid. An aliquot (0.2 ml) of the dissolved material was then added to 9.8 ml of Aquasol and counted in a scintillation counter.

15 Et kalcifikationsforhold blev beregnet for de forbindelser, der er af interesse, og blev bestemt som følger: 1,25 (OH)2D3 cpm - kontrol cpmA calcification ratio was calculated for the compounds of interest and was determined as follows: 1.25 (OH) 2D3 cpm - control cpm

Kalcifikationsforhold = -- forbindelsens cpm - kontrol cpm 20 De fundne kalcifikationsforhold for forskellige vitamin D-analoger, administreret ved de to administrationsveje, er som følger:Calcification ratios = - compound cpm - control cpm 20 Calcification ratios for different vitamin D analogues, administered by the two routes of administration, are as follows:

Forbindelse Subkutan Topisk 1.25 (OH)2D3 XI 1 25 1,25 (ΟΗ)2-16ΔΌ3 A 486 25 (ΟΗ)-16ΔΒ3 B 5 1.25 (OH)2-16A-23-ynD3 Ξ 47 <1 25 (OH)-16A-23-ynD3 F >1400 >34 25 (OH)-16A-23-en-D3 D >1400 >34 30 ___________Compound Subcutaneous Topical 1.25 (OH) 2D3 XI 1 25 1.25 (ΟΗ) 2-16ΔΌ3 A 486 25 (ΟΗ) -16ΔΒ3 B 5 1.25 (OH) 2-16A-23-ynD3 Ξ 47 <1 25 (OH) - 16A-23-inD3 F> 1400> 34 25 (OH) -16A-23-and-D3 D> 1400> 34 30 ___________

Ovenstående data viser, at forbindelserne ifølge opfindelsen ved subkutan administration forårsager mindre kalcifikation af blødt væv end 12 DK 169945 B1 den, forbindelse X forårsager. Kalcifikation af blødt væv er en uønsket bivirkning for en forbindelse, der anvendes til behandling af hyperproliferative hudsygdomme og neoplastiske sygdomme.The above data shows that, by subcutaneous administration, the compounds of the invention cause less soft tissue calcification than the compound X causes. Soft tissue calcification is an undesirable side effect of a compound used to treat hyperproliferative skin diseases and neoplastic diseases.

5 Aktiviteten af forbindelser med formlen I som midler til behandling af hyperproliferative hudsygdomme kan påvises fx ved testmetoder, der er kendte inden for teknikken, således som det er angivet i The Society for Investigative Dermatology 1986, 709-714. Virkningerne af de ovennævnte forbindelser A til F på den morfologiske differentie-10 ring af dyrkede humane keratinocytter sammenlignet med virkningen af 1,25-dihydroxycholecalciferol (forbindelse X) er i nedenstående tabeller 1-4 udtrykt som antallet (xlO4) af dyrkede humane keratinocytter. En forbindelse, som inducerer differentieringen af basal-celler til pladeepithelceller og dækceller ("envelope cells"), er 15 nyttig som et middel til behandling af hudsygdomme, der er karakteriseret ved keratiniseringslidelser såsom psoriasis.The activity of compounds of formula I as agents for the treatment of hyperproliferative skin diseases can be demonstrated, for example, by test methods known in the art, as disclosed in The Society for Investigative Dermatology 1986, 709-714. The effects of the above compounds A to F on the morphological differentiation of cultured human keratinocytes compared to the action of 1,25-dihydroxycholecalciferol (Compound X) are expressed in Tables 1-4 below as the number (x104) of cultured human keratinocytes. A compound which induces the differentiation of basal cells into squamous cell and envelope cells is useful as an agent for the treatment of skin diseases characterized by keratinization disorders such as psoriasis.

Tabel 1Table 1

Forbindelse Dosis Antal celler (xlO4): (M) plade- 20 total basal- epithel- dækceller celler cellerCompound Dose Number of cells (x104): (M) plate total basal epithelial cover cells cells cells

Kontrol 133±5 118±4 15±1 18±2 X 10'10 122+4 103±2 19±2 23±1 25 10'8 112+6 89+2 23±4 30±3 10"6 95±7 64±6 31+1 34±2 A 10'10 132+8 115±7 17±1 27±2 10"8 128±10 106+8 22±2 33±2 10"6 101±7 71±5 30±2 39±2 30 B 10'10 133±6 115±5 18±1 25±1 10"8 131±4 109±2 22±2 29±2 10'6 104±4 74±3 30±1 33±1Control 133 ± 5 118 ± 4 15 ± 1 18 ± 2 X 10'10 122 + 4 103 ± 2 19 ± 2 23 ± 1 25 10'8 112 + 6 89 + 2 23 ± 4 30 ± 3 10 "6 95 ± 7 64 ± 6 31 + 1 34 ± 2 A 10'10 132 + 8 115 ± 7 17 ± 1 27 ± 2 10 "8 128 ± 10 106 + 8 22 ± 2 33 ± 2 10" 6 101 ± 7 71 ± 5 30 ± 2 39 ± 2 30 B 10'10 133 ± 6 115 ± 5 18 ± 1 25 ± 1 10 "8 131 ± 4 109 ± 2 22 ± 2 29 ± 2 10'6 104 ± 4 74 ± 3 30 ± 1 33 ± 1

Tabel 2 DK 169945 Bl 13Table 2 DK 169945 Pg 13

Forbindelse Dosis Antal celler (xlO4): (M) plade- total basal- epithel- dæk- 5 celler celler cellerCompound Dose Number of cells (x104): (M) plate total basal epithelial cover cells cells cells

Kontrol 123+7 105±6 18±1 74±7 X 10'10 116+9 95±8 21±1 91±4 10'8 101±10 75+8 26+2 122±11 10 10'6 83+5 57+4 26±1 146±16 C 10'10 117±4 92±2 25±2 103±6 10"8 108+3 80±2 28+1 128+3 10‘6 80±7 54±6 26+1 153±1 D 10'10 113±7 93±6 20,1 104±10 15 10'8 111+7 86±3 25±2 128±5 10'6 94+3 68±1 26±2 144±7Control 123 + 7 105 ± 6 18 ± 1 74 ± 7 X 10'10 116 + 9 95 ± 8 21 ± 1 91 ± 4 10'8 101 ± 10 75 + 8 26 + 2 122 ± 11 10 10'6 83+ 5 57 + 4 26 ± 1 146 ± 16 C 10'10 117 ± 4 92 ± 2 25 ± 2 103 ± 6 10 "8 108 + 3 80 ± 2 28 + 1 128 + 3 10'6 80 ± 7 54 ± 6 26 + 1 153 ± 1 D 10'10 113 ± 7 93 ± 6 20.1 104 ± 10 15 10'8 111 + 7 86 ± 3 25 ± 2 128 ± 5 10'6 94 + 3 68 ± 1 26 ± 2 144 ± 7

Tabel 3Table 3

Forbindelse Dosis Antal celler (xlO4): 20 (M) plade- total basal- epithel- dækceller celler cellerCompound Dose Number of cells (x104): 20 (M) plate total basal epithelial cover cells cells cells

Kontrol 108±10 93±8 15±2 88±8 25 X 10'10 106±7 86+6 18+1 100+9 10'8 84+8 61+5 23+3 122+8 10'6 73±7 51+5 22±2 142±11 E 10'10 86±4 63±2 23±2 114±5 10'8 82±3 53+2 29+1 141±5 30 10'6 78+3 41±1 27±2 147±4 F 10'10 103+5 81+3 22±2 103±4 10'8 97+3 67±2 29±1 121±6 10'6 84+4 55±2 29±1 137±7 35 Aktiviteten af forbindelser med formlen 1 som midler til behandling af hyperproliferative hudsygdomme kan også påvises ved at bestemme antallet af dyrkede humane keratinocytter og antallet af dannede 14 DK 169945 B1 dækceller samt antallet af dyrkede pladeepithelcarcinom-cellelinier (SCC-115) i nærværelse af de nævnte forbindelser. Resultaterne er vist i tabel 4 og 5:Control 108 ± 10 93 ± 8 15 ± 2 88 ± 8 25 X 10'10 106 ± 7 86 + 6 18 + 1 100 + 9 10'8 84 + 8 61 + 5 23 + 3 122 + 8 10'6 73 ± 7 51 + 5 22 ± 2 142 ± 11 E 10'10 86 ± 4 63 ± 2 23 ± 2 114 ± 5 10'8 82 ± 3 53 + 2 29 + 1 141 ± 5 30 10'6 78 + 3 41 ± 1 27 ± 2 147 ± 4 F 10'10 103 + 5 81 + 3 22 ± 2 103 ± 4 10'8 97 + 3 67 ± 2 29 ± 1 121 ± 6 10'6 84 + 4 55 ± 2 29 ± 1 137 ± 7 35 The activity of compounds of formula 1 as agents for the treatment of hyperproliferative skin diseases can also be demonstrated by determining the number of cultured human keratinocytes and the number of cultured squamous cell carcinoma cell lines (SCC-115) in presence of said compounds. The results are shown in Tables 4 and 5:

Tabel 4 5 Behandling Dosis Antal Antal (M) keratinocytter dækceller (xlO4) (xlO2)Table 4 5 Treatment Dose Number Number (M) keratinocytes tire cells (x104) (x102)

Kontrol 189,49+22,3 858,28±185,70 10 (0,1% ethanol)Control 189.49 + 22.3 858.28 ± 185.70 (0.1% ethanol)

Forbindelse E 10'12 187,36+15,33 1136,63± 383,66 10"10 175,34+10,19 1444,87± 312,47 10'8 145,79±15,66 2113,62+1049,33 10'6 41,95± 7,53 1916,83+ 887,66 15 Kontrol 148,73±16,23 2193,7± 921,9 (0,1% ethanol)Compound E 10'12 187.36 + 15.33 1136.63 ± 383.66 10 "10 175.34 + 10.19 1444.87 ± 312.47 10'8 145.79 ± 15.66 2113.62+ 1049.33 10'6 41.95 ± 7.53 1916.83 + 887.66 Control 148.73 ± 16.23 2193.7 ± 921.9 (0.1% ethanol)

Forbindelse A 10'12 114,91±10,95 1662,2±420,1 10'10 130,37+24,32 3973,8+126,99 10'8 120,67+16,87 7235,2+ 55,5 20 10"6 109,22+15,87 8323,5+157,6Compound A 10'12 114.91 ± 10.95 1662.2 ± 420.1 10'10 130.37 + 24.32 3973.8 + 126.99 10'8 120.67 + 16.87 7235.2+ 55.5 20 10 "6 109.22 + 15.87 8323.5 + 157.6

Tabel 5Table 5

Behandling Dosis Antal (M) cellelinier (SCC-15) 25 (xlO"5)Treatment Dose Number of (M) cell lines (SCC-15) 25 (x10 5)

Kontrol 7,35+1,75Control 7.35 + 1.75

Forbindelse E 10'12 6,98+1,68 10'10 5,89+1,58 30 10'8 5,76+1,53 10'6 0,40+0,98Compound E 10'12 6.98 + 1.68 10'10 5.89 + 1.58 30 10'8 5.76 + 1.53 10'6 0.40 + 0.98

Forbindelse A 10"6 0,49+0,13 15 DK 169945 B1Compound A 10 "6 0.49 + 0.13 DK 169945 B1

Ovenstående resultater viser, at forbindelserne med formlen I inducerer differentiering af hudceller, og de er således nyttige til behandling af hyperproliferative hudsygdomme såsom psoriasis.The above results show that the compounds of formula I induce differentiation of skin cells and are thus useful in the treatment of hyperproliferative skin diseases such as psoriasis.

For at påvise aktiviteten af forbindelserne med formlen I som midler 5 til behandling af neoplastiske sygdomme blev de anti-proliferative (AP) og de differentierings-inducerende (DI) virkninger af forbindelserne A til F og humane promyelocytiske HL-60 tumorceller vurderet.To demonstrate the activity of the compounds of formula I as agents 5 for the treatment of neoplastic diseases, the anti-proliferative (AP) and differentiation-inducing (DI) effects of compounds A to F and human promyelocytic HL-60 tumor cells were assessed.

I tabel 6 er AP-virkningen angivet som procentvis reduktion af celleantal og som koncentration ID50 af forbindelsen, hvilken koncentra-10 tion reducerede celleantallet med 50%. DI-virkningen udtrykkes som procent differentierede celler og som koncentrationen ED50 af forbindelser, hvilken koncentration inducerede en 50%'s differentiering af cellerne.In Table 6, the AP effect is given as a percentage reduction in cell number and as concentration ID 50 of the compound, which concentration reduced the cell number by 50%. The DI effect is expressed as percent differentiated cells and as the concentration of ED50 of compounds, which induced a 50% differentiation of the cells.

Tabel 6 15 Koncentration % Reduktion ID5q Differen- ED50Table 6 Concentration% Reduction ID5q Differen- ED50

x10"8M) i celleantal (xl0"8M) tierede (xlO'8Mx10 "8M) in cell count (x10" 8M) tiered (x10 "8M

celler(%)cells (%)

Forbindelse XCompound X

20 0,01 6 3 0,1 5 11 1 16 2 19 2 10 66 68 100 84 9820 0.01 6 3 0.1 5 11 1 16 2 19 2 10 66 68 100 84 98

25 Forbindelse ACompound A

0,01 10 3 0,1 33 16 1 84 0,2 92 0,2 10 85 97 30 100 85 98 16 DK 169945 B10.01 10 3 0.1 33 16 1 84 0.2 92 0.2 10 85 97 30 100 85 98 16 DK 169945 B1

Forbindelse BCompound B

0,1 10 5 18 4 10 14 35 6 32 5 100 82 93 1000 95 950.1 10 5 18 4 10 14 35 6 32 5 100 82 93 1000 95 95

Forbindelse CCompound C

0,01 18 3 0,1 20 19 10 1 81 0,3 92 0,3 10 85 97 100 86 990.01 18 3 0.1 20 19 10 1 81 0.3 92 0.3 10 85 97 100 86 99

Forbindelse DCompound D

0,1 12 1 15 1 12 2 10 17 150 17 200 100 46 31 1000 95 970.1 12 1 15 1 12 2 10 17 150 17 200 100 46 31 1000 95 97

Forbindelse ECompound E

20 0,01 6 9 0,1 59 50 1 80 0,07 96 0,1 10 81 9820 0.01 6 9 0.1 59 50 1 80 0.07 96 0.1 10 81 98

Forbindelse FCompound F

25 0,1 13 4 1 10 12 10 8 70 21 70 100 58 55 1000 95 91 30 Disse data viser, at hver af de pågældende forbindelser hæmmer pro-liferationen af humane promyelocytiske celler in vitro, selvom de ikke er toksiske overfor cellerne. Endvidere differentierer cellerne mod en mere moden phenotype ved de samme doser, som inhiberer proliferation. Ud fra disse resultater ses, at hver af de testede forbin-35 delser er nyttige som et middel til behandling af neoplastiske sygdomme såsom leukæmi.25 0.1 13 4 1 10 12 10 8 70 21 70 100 58 55 1000 95 91 30 These data show that each of the compounds in question inhibits the proliferation of human promyelocytic cells in vitro, although not toxic to the cells. Furthermore, the cells differentiate into a more mature phenotype at the same doses that inhibit proliferation. From these results, it is seen that each of the compounds tested is useful as a remedy for treating neoplastic diseases such as leukemia.

17 DK 169945 B117 DK 169945 B1

Forbindelserne med formlen I kan administreres oralt til behandling af neoplastiske sygdomme eller til behandling af hyperproliferative hudsygdomme hos varmblodige dyr, der behøver en sådan behandling, fx til et voksent menneske, i doser, der ligger i området fra ca. 0,1 5 til 10 μg pr. dag.The compounds of formula I may be administered orally for the treatment of neoplastic diseases or for the treatment of hyperproliferative skin diseases in warm-blooded animals in need of such treatment, for example to an adult, in doses ranging from 0.1 to 10 µg per day.

Til behandling af hyperproliferative hudsygdomme kan forbindelserne med formlen I også administreres topisk til varmblodige dyr, som behøver en sådan behandling, i doser på ca. 1 til 1000 pg pr. gram topisk formulering pr. dag.For the treatment of hyperproliferative skin diseases, the compounds of Formula I may also be administered topically to warm-blooded animals in need of such treatment, at doses of ca. 1 to 1000 pg per grams of topical formulation per day.

10 Orale doseringsformer omfattende forbindelser med formlen I kan inkorporeres i fx kapsler eller tabletter sammen med farmaceutisk acceptable bærere. Eksempler på sådanne bærematerialer, som kan inkorporeres i kapsler, er bindemidler såsom gummi traganth eller gelatine; excipienser såsom dicalciumphosphat; desintegrationsmidler 15 såsom majsstivelse; glittemidler såsom magnesiumstearat; sødemidler såsom sucrose; smagsgivende midler såsom pebermynte. Tabletter kan overtrækkes med schellak, sukker eller begge dele. En sirup eller eliksir kan indeholde et sødemiddel, methyl- og propylparabener som konserveringsmidler, et farvestof og et smagsgivende middel.Oral dosage forms comprising compounds of formula I may be incorporated into, for example, capsules or tablets, together with pharmaceutically acceptable carriers. Examples of such carriers which can be incorporated into capsules are binders such as gum tragacanth or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch; lubricants such as magnesium stearate; sweeteners such as sucrose; flavoring agents such as peppermint. Tablets can be coated with shellac, sugar or both. A syrup or elixir may contain a sweetener, methyl and propyl parabens as preservatives, a dye and a flavoring agent.

20 Topiske doseringsformer omfattende forbindelser med formlen I omfatter salver og cremer, herunder formuleringer med olieagtige, adsorberbare, vandopløselige og emulsions-type baser såsom lanolin og polyethylenglycoler. Topiske doseringsformer omfatter også geler, lotioner, pulvere og aerosoler. De topiske præparater kan også an-25 vendes til behandling af hudinflammationer eller af slimhinder, fx slimhinden i munden eller i nedre colon.Topical dosage forms comprising compounds of formula I include ointments and creams, including formulations with oily, adsorbable, water-soluble and emulsion-type bases such as lanolin and polyethylene glycols. Topical dosage forms also include gels, lotions, powders and aerosols. The topical compositions may also be used to treat skin inflammation or mucous membranes, for example, the mucosa of the mouth or lower colon.

Lotioner, dvs. flydende præparater, der varierer fra simple opløsninger til vandige eller hydroalkoholiske præparater indeholdende fint opdelte substanser, kan indeholde suspensions- eller disperge-30 ringsmidler såsom cellulosederivater, fx ethyl- eller methylcellu-lose; gelatine eller gummier, der indeholder den aktive bestanddel i en bærer fremstillet af vand, alkohol eller glycerin. Geler er halvfaste præparater fremstillet ved at gelere en opløsning eller suspension af den aktive bestanddel i en bærer. Bæreme, som kan være vand- DK 169945 B1Lotions, ie liquid compositions ranging from simple solutions to aqueous or hydroalcoholic preparations containing finely divided substances may contain suspending or dispersing agents such as cellulose derivatives, e.g., ethyl or methyl cellulose; gelatin or gums containing the active ingredient in a carrier made of water, alcohol or glycerine. Gels are semi-solid preparations prepared by gelling a solution or suspension of the active ingredient in a carrier. The carriers which may be water DK 169945 B1

ISICE

holdige eller vandfri, geleres under anvendelse af et geleringsmiddel, fx carboxypolymethylen, og neutraliseres til passende gelkonsistens under anvendelse af alkali, fx natriumhydroxid, eller aminer såsom polyethylencocoamin.containing or anhydrous, are gelled using a gelling agent, e.g., carboxypolymethylene, and neutralized to appropriate gel consistency using alkali, e.g., sodium hydroxide, or amines such as polyethylene cocoamine.

5 EKSEMPEL 1 a) En blanding af 3,24 g [l(R*),3aR*-(3a)S,4a,7aa)]-3a,4,5,6,7,7a-hexahydro-4-hydroxy-/3,7a-dimethyl-3H-inden-1-ethanol, 30 ml pyridin og 3,51 g p-toluensulfonylchlorid omrøres ved 0° i 18 timer. Efter tilsætning af is og fortynding med vand ekstraheres blandingen med 10 methylenchlorid. Den organiske fase vaskes med IN H2S04, mættetEXAMPLE 1 a) A mixture of 3.24 g [1 (R *), 3aR * - (3a) S, 4a, 7aa)] - 3a, 4,5,6,7,7a-hexahydro-4-hydroxy - / 3,7a-Dimethyl-3H-indene-1-ethanol, 30 ml of pyridine and 3.51 g of p-toluenesulfonyl chloride are stirred at 0 ° for 18 hours. After adding ice and diluting with water, the mixture is extracted with 10 methylene chloride. The organic phase is washed with 1N H2SO4, saturated

NaHC03, tørres derefter og inddampes. Remanensen kromatograferes på silicagel med ethylacetat-hexan (1:1:5) til opnåelse af 4,61 g (82%) [1 (R*) , 3aR*- (3a/3,4a, 7aa) ] -3a, 4,5,6,7,7a-hexahydro-4-hydroxy-/?, 7a-dimethyl-3H-inden-l-ethanol-4-methylbenzensulfonat, [a]^ = + 31,9° 15 (c = 0,53, CHC13).NaHCO3, then dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1: 1: 5) to give 4.61 g (82%) [1 (R *), 3aR * - (3a / 3.4a, 7aa)] -3a, 4 , 5,6,7,7a-hexahydro-4-hydroxy-β, 7a-dimethyl-3H-indene-1-ethanol-4-methylbenzenesulfonate, [α] D = + 31.9 ° (c = 0, 53, CHCl3).

b) Til en opløsning af 4,61 g af det i a) vundne produkt i 22 ml DMSO tilsættes 1,10 g natriumcyanid, og blandingen opvarmes ved 90°C i 2 timer. Efter afkøling til stuetemperatur bringes blandingen tinder formindsket tryk til fjernelse af opløsningsmidlet og fortyndes 20 derefter med vand. Blandingen ekstraheres med ether. Den organiske fase vaskes med mættet saltvand, tørres og inddampes. Remanensen kromatograferes på silicagel med methylenchlorid-hexan-ethylacetat (86:7:7) til opnåelse af 2,52 g (91%) [1 (R*) ,3aR*-(3aj8,4a,7aa) ] -3a,4,5,6,7,7a-hexahydro-4-hydroxy-/?, 7a-dimethyl-3H-inden-l-propan- 25 nitril, [aig1 = + 29,2° (c = 0,65, CHCI3).b) To a solution of 4.61 g of the product obtained in (a) in 22 ml of DMSO is added 1.10 g of sodium cyanide and the mixture is heated at 90 ° C for 2 hours. After cooling to room temperature, the mixture is brought to a reduced pressure to remove the solvent and then diluted with water. The mixture is extracted with ether. The organic phase is washed with saturated brine, dried and evaporated. The residue is chromatographed on silica gel with methylene chloride-hexane-ethyl acetate (86: 7: 7) to give 2.52 g (91%) [1 (R *), 3aR * - (3a8.4a, 7aa)] -3a, 4 , 5,6,7,7a-Hexahydro-4-hydroxy-β, 7a-dimethyl-3H-indene-1-propanitrile, [aig 1 = + 29.2 ° (c = 0.65, CHCl 3) .

c) Til en blanding af 6,85 ml diisobutylaluminiumhydrid i hexan og 5,2 ml methylenchlorid ved -6°C tilsættes en opløsning af 0,430 g af det i b) vundne produkt i 10 ml methylenchlorid. Blandingen omrøres ved -6°C i 55 minutter. Efter tilsætning af mættet ammoniumchlorid 30 hydrolyseres blandingen med 3N HC1-ether (2:1). Den vandige fase * ekstraheres med ether. De organiske faser vaskes med mættet saltvand, tørres og inddampes. Remanensen kromatograferes på silicagel med ethylacetat-hexan (1:2) til opnåelse af 260 mg (60%) [1(R*),3a/?*- 19 DK 169945 B1 (3a/3,4α,7aa)]-3a,4,5,6,7,7a-hexahydro-4-hydroxy-/?,7a-dimethyl-3H-inden-l-propanal, [a]p2 = + 43,1° (c = 0,32, CHCI3) .c) To a mixture of 6.85 ml of diisobutyl aluminum hydride in hexane and 5.2 ml of methylene chloride at -6 ° C is added a solution of 0.430 g of the product obtained in (b) in 10 ml of methylene chloride. The mixture is stirred at -6 ° C for 55 minutes. After addition of saturated ammonium chloride, the mixture is hydrolyzed with 3N HCl ether (2: 1). The aqueous phase * is extracted with ether. The organic phases are washed with saturated brine, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1: 2) to give 260 mg (60%) [1 (R *), 3a /? * - 3a / 3.4a, 7aa)] - 3a , 4,5,6,7,7a-Hexahydro-4-hydroxy-β, 7α-dimethyl-3H-indene-1-propanal, [α] β 2 = + 43.1 ° (c = 0.32, CHCl 3 ).

d) En blanding af 1,77 g triphenylphosphin, 2,23 g carbontetrabromid, 441 mg zinkstøv og 23 ml methylenchlorid omrøres ved 25°C i 31 timer.d) A mixture of 1.77 g of triphenylphosphine, 2.23 g of carbon tetrabromide, 441 mg of zinc dust and 23 ml of methylene chloride is stirred at 25 ° C for 31 hours.

5 Til denne blanding tilsættes en opløsning af 0,430 g af det i c) vundne produkt i 38 ml methylenchlorid, og blandingen omrøres i 18 timer. Blandingen fortyndes med pentan, og uopløseligt materiale frafiltreres. Den uopløselige fraktion opløses i methylenchlorid, og opløsningen fortyndes igen med pentan. Efter filtrering inddampes de 10 samlede filtrater. Remanensen oprenses på silicagel med 1:4 ethyl- acetat-hexan til opnåelse af 0,490 g (67%) [1 (R*) ,3aR*-(3a/J,4a,7aa) ] -1-(4,4-dibrom-1-methy1-3 -buteny1)-3a,4,5,6,7,7a-hexahydro-7a-methyl-3H-inden-4-ol, [a]g2 = + 14,4° (c = 0,55, CHCI3).To this mixture is added a solution of 0.430 g of the product obtained in (c) in 38 ml of methylene chloride and the mixture is stirred for 18 hours. The mixture is diluted with pentane and the insoluble material is filtered off. The insoluble fraction is dissolved in methylene chloride and the solution is again diluted with pentane. After filtration, the 10 total filtrates are evaporated. The residue is purified on silica gel with 1: 4 ethyl acetate-hexane to give 0.490 g (67%) [1 (R *), 3aR * - (3a / J, 4a, 7aa)] -1- (4.4- dibromo-1-methyl-3-butyl] -3a, 4,5,6,7,7a-hexahydro-7a-methyl-3H-inden-4-ol, [a] g2 = + 14.4 ° (c = 0.55, CHCl 3).

e) Til en opløsning af 0,680 g af det i d) vundne produkt i 31 ml THF 15 ved -75°C tilsættes dråbevis 3,77 ml 1,6M opløsning af butyllithium i hexan. Blandingen omrøres ved -75°C i 1 time og ved 25°C i 1 time.e) To a solution of 0.680 g of the product obtained in d) in 31 ml of THF 15 at -75 ° C is added dropwise 3.77 ml of 1.6M solution of butyllithium in hexane. The mixture is stirred at -75 ° C for 1 hour and at 25 ° C for 1 hour.

Efter tilsætning af mættet saltvand fortyndes blandingen med mættet vandig NaHCC>3 og ekstraheres med ether. Den organiske fase vaskes med mættet saltvand, tørres og inddampes. Remanensen kromatograferes på 20 silicagel med ethylacetat-hexan (1:4) til opnåelse af 0,350 g (89%) [1(R*) ,3aR*-(3a^,4a,7aa)]-3a,4,5,6,7,7a-hexahydro-7a-methyl-l-(1-methyl-3-butynyl)-3H-inden-4-ol, [α]β2 = + 30,7° (c = 0,42, CHCI3).After addition of saturated saline, the mixture is diluted with saturated aqueous NaHCC> 3 and extracted with ether. The organic phase is washed with saturated brine, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1: 4) to give 0.350 g (89%) [1 (R *), 3aR * - (3a ^, 4a, 7aa)] - 3a, 4,5.6 , 7,7a-Hexahydro-7a-methyl-1- (1-methyl-3-butynyl) -3H-inden-4-ol, [α] β2 = + 30.7 ° (c = 0.42, CHCl3) .

f) Til en opløsning af 1,29 g af det i e) vundne produkt i 80 ml methylenchlorid tilsættes 3,59 g 1-(trimethylsilyl)imidazol. Blandin- 25 gen omrøres ved 25°C i 3 timer. Efter tilsætning af 40 ml vand og omrøring i 20 minutter ekstraheres blandingen med ethylacetat. Den organiske fase vaskes med vand og mættet saltvand, tørres og inddampes. Remanensen oprenses på silicagel med ethylacetat-hexan (1:15) til opnåelse af 1,70 g (99%) [1 (R*) ,3aR*- (3a/?,4a, 7aa) ] -3a,4,5, 6,7, 7a- 30 hexahydro-7a-methyl-l-(1-methyl-3-butynyl)-4-[(trimethylsilyl)oxy]- 3H-inden, [a]g° = + 39,7“ (c = 0,30, CHCI3).f) To a solution of 1.29 g of the product obtained in e) in 80 ml of methylene chloride is added 3.59 g of 1- (trimethylsilyl) imidazole. The mixture is stirred at 25 ° C for 3 hours. After adding 40 ml of water and stirring for 20 minutes, the mixture is extracted with ethyl acetate. The organic phase is washed with water and saturated brine, dried and evaporated. The residue is purified on silica gel with ethyl acetate-hexane (1:15) to give 1.70 g (99%) of [1 (R *), 3aR * - (3a / 4a, 7aa)] -3a, 4.5 , 6,7, 7a-hexahydro-7a-methyl-1- (1-methyl-3-butynyl) -4 - [(trimethylsilyl) oxy] -3H-indene, [a] g ° = + 39.7 (c = 0.30, CHCl 3).

g) Til en opløsning af 1,70 g af det i f) vundne produkt i 48 ml THF ved -75°C tilsættes dråbevis 6,01 ml 1,6M butyllithium i hexan. Efter omrøring i 40 minutter tilsættes 3,05 ml acetone, og blandingen omrø- 20 DK 169945 B1 res ved -75°C i 20 minutter og ved 25°C i 75 minutter. Efter tilsætning af 40 ml af en 1:1 blanding af 2M KHCO3 og 1M kaliumnatriumtartrat omrøres blandingen i 20 minutter og ekstraheres derefter med ethylacetat. Den organiske fase vaskes med mættet saltvand, tørres og 5 inddampes. Remanensen kromatograferes på silicagel med ethylacetat-hexan (1:5) til opnåelse af 1,62 g (89%) [1 (R*), 3aR*-(3ay3,4a, 7aa) ]- 6-(3a,4,5,6,7,7a-hexahydro-7a-methyl-4-[(trimethylsilyl)oxy]-3H-inden-l-yl)-2-methyl-3-heptyn-l-ol, [α]β° = + 39,7° (c = 0,30, CHCI3).g) To a solution of 1.70 g of the product obtained in f) in 48 ml of THF at -75 ° C is added dropwise 6.01 ml of 1.6M butyllithium in hexane. After stirring for 40 minutes, 3.05 ml of acetone is added and the mixture is stirred at -75 ° C for 20 minutes and at 25 ° C for 75 minutes. After adding 40 ml of a 1: 1 mixture of 2M KHCO3 and 1M potassium sodium tartrate, the mixture is stirred for 20 minutes and then extracted with ethyl acetate. The organic phase is washed with saturated brine, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1: 5) to give 1.62 g (89%) [1 (R *), 3aR * - (3a3.4a, 7aa)] - 6- (3a, 4, 5,6,7,7a-hexahydro-7a-methyl-4 - [(trimethylsilyl) oxy] -3H-inden-1-yl) -2-methyl-3-heptyn-1-ol, [α] β ° = + 39.7 ° (c = 0.30, CHCl 3).

10 h) Til en opløsning af 1,62 g af det i g) vundne produkt i 53 ml THF tilsættes 15,5 ml 1M tetrabutylammoniumfluorid i THF. Blandingen omrøres i 50 minutter. Efter fortynding med halvmættet NaHCC^ inddampes blandingen til fjernelse af det meste af opløsningsmidlet og ekstraheres med ethylacetat. Den organiske fase vaskes med halvmættet 15 saltvand, tørres og inddampes. Remanensen kromatograferes på silicagel med ethylacetat-hexan (1:1) til opnåelse af 1,17 g (82%) [1 (R*), -3aR*- (3ay8,4a,7aot) ] -3a,4,5,6,7,7a-hexahydro-l- (1,5-dimethyl-5-hydroxy-3-hexynyl)-7a-methyl-3H-inden-4-ol, smp. 105-107®.10 h) To a solution of 1.62 g of the product obtained in g) in 53 ml of THF is added 15.5 ml of 1M tetrabutylammonium fluoride in THF. The mixture is stirred for 50 minutes. After dilution with semi-saturated NaHCC 3, the mixture is evaporated to remove most of the solvent and extracted with ethyl acetate. The organic phase is washed with half-saturated brine, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1: 1) to give 1.17 g (82%) [1 (R *), -3aR * - (3a8.4a, 7aot)] -3a, 4.5, 6,7,7a-hexahydro-1- (1,5-dimethyl-5-hydroxy-3-hexynyl) -7a-methyl-3H-inden-4-ol, m.p. 105-107®.

i) Til en opløsning af 0,720 g af det i h) vundne produkt i 44 ml 20 methylenchlorid tilsættes 1,59 g natriumacetat og 3,18 g 2,2'-bi- pyridiniumchlorchromat. Blandingen omrøres i 2 timer. Yderligere 1,59 g 2,2'-bipyridiniumchlorchromat tilsættes derefter, og omrøringen fortsættes i 2 timer. Efter tilsætning af 6 ml 2-propanol fortyndes blandingen derefter med vand og ekstraheres med ether-ethyl-25 acetat (1:1) . Den organiske fase vaskes med vand, IN H2S04, mættet NaHC03 og mættet saltvand. Efter tørring inddampes opløsningen, og remanensen kromatograferes på silicagel med ethylacetat-hexan (1:1) til opnåelse af 0,560 g (78%) [1 (R*) ,3aR*- (3a/0,7aa)] -3,3a,5,6,7,7a-hexahydro-1-(5-hydroxy-1,5-dimethyl-3-hexynyl)-7a-methyl-4H-inden-30 4-on, [a]£° = +35,3° (c - 0,36, CHCI3).i) To a solution of 0.720 g of the product obtained in h) in 44 ml of methylene chloride is added 1.59 g of sodium acetate and 3.18 g of 2,2'-bipyridinium chlorochromate. The mixture is stirred for 2 hours. An additional 1.59 g of 2,2'-bipyridinium chlorochromate is then added and stirring is continued for 2 hours. After adding 6 ml of 2-propanol, the mixture is then diluted with water and extracted with ether-ethyl acetate (1: 1). The organic phase is washed with water, 1N H2SO4, saturated NaHCO3 and saturated brine. After drying, the solution is evaporated and the residue is chromatographed on silica gel with ethyl acetate-hexane (1: 1) to give 0.560 g (78%) [1 (R *), 3aR * - (3a / 0.7aa)] -3.3a , 5,6,7,7a-Hexahydro-1- (5-hydroxy-1,5-dimethyl-3-hexynyl) -7a-methyl-4H-inden-4-one, [a] + = +35 , 3 ° (c - 0.36, CHCl 3).

j) Til en opløsning af 0,552 g af det i i) vundne produkt i 70 ml methylenchlorid tilsættes 2,00 g 1-(trimethylsilyl)imidazol. Efter omrøring i 17 timer og tilsætning af 22 ml vand ekstraheres blandingen med ethylacetat. Den organiske fase vaskes med vand og mættet 35 saltvand, tørres derefter og inddampes. Remanensen kromatograferes på DK 169945 Bl 21 silicagel med ethylacetat-hexan (1:4) til opnåelse af 0,693 g (99%) [1 (R*) ,3aR*- (3a/J,7act)] -3,3a,5,6,7,7a-hexahydro-l- (l,5-dimethyl-5-[(trimethylsilyl)oxy]-3-hexynyl)-7a-methyl-4H-inden-4-on, = + 29,5° (c = 0,20, CHCI3).j) To a solution of 0.552 g of the product obtained in i) in 70 ml of methylene chloride is added 2.00 g of 1- (trimethylsilyl) imidazole. After stirring for 17 hours and adding 22 ml of water, the mixture is extracted with ethyl acetate. The organic phase is washed with water and saturated brine, then dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1: 4) to give 0.693 g (99%) [1 (R *), 3aR * - (3a / J, 7act)] -3.3a, 5 , 6,7,7a-hexahydro-1- (1,5-dimethyl-5 - [(trimethylsilyl) oxy] -3-hexynyl) -7a-methyl-4H-inden-4-one, = + 29.5 ° (c = 0.20, CHCl 3).

5 k) Til en opløsning af 2,00 g [3S-(1Ζ,3α,5^)]-[2-[3,5-bis [ [ (1,ldi-methyl) dimethylsilyl] oxy] -2-methylencyclohexyliden] ethyl] diphenyl-phosphinoxid i 45 ml THF ved -75°C tilsættes dråbevis 1,87 ml 1,6M butyllithium i hexan. Efter omrøring i 6 minutter tilsættes dråbevis en opløsning af 0,693 g af det i j) vundne produkt i 26 ml THF. Efter 10 omrøring ved -75°C i 70 minutter og tilsætning af en 1:1 blanding af 1M kaliumnatriumtartrat og 2M KHC03 ekstraheres blandingen med ethyl-acetat. Den organiske fase vaskes med mættet saltvand, tørres og inddampes. Remanensen kromatograferes på silicagel med ethylacetat-hexan (1:15) til opnåelse af 1,23 g (87%) (la,30,5Z,7E)-1,3-bis[[1,1-15 dimethylethyl)dimethylsilyl]-oxy-25-[(trimethylsilyl)oxy]-9,10-seco- cholesta-5,7,10(19),16-tetraen-3-yn, [a]^3 = + 47,1° (c = 0,21, CHCI3).K) To a solution of 2.00 g of [3S- (1Ζ, 3α, 5β)] - [2- [3,5-bis [[(1,1di-methyl) dimethylsilyl] oxy] -2-methylene cyclohexylidene ] ethyl] diphenylphosphine oxide in 45 ml of THF at -75 ° C is added dropwise 1.87 ml of 1.6M butyllithium in hexane. After stirring for 6 minutes, a solution of 0.693 g of the product obtained in j) in 26 ml of THF is added dropwise. After stirring at -75 ° C for 70 minutes and adding a 1: 1 mixture of 1M potassium sodium tartrate and 2M KHCO3, the mixture is extracted with ethyl acetate. The organic phase is washed with saturated brine, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:15) to give 1.23 g (87%) (Ia, 30.5Z, 7E) -1,3-bis [[1,1-15 dimethylethyl) dimethylsilyl] -oxy-25 - [(trimethylsilyl) oxy] -9,10-seco-cholesta-5,7,10 (19), 16-tetraene-3-yn, [α] 3 = + 47.1 ° (c = 0.21, CHCl 3).

1) Til en opløsning af 0,228 g af det i k) vundne produkt i 11 ml THF tilsættes 1,92 ml 1M tetrabutylammoniumfluorid i THF. Blandingen 20 omrøres i 16 timer. Efter fortynding med vand ekstraheres blandingen med ethylacetat. Den organiske fase vaskes med halvmættet saltvand og mættet saltvand, tørres og inddampes. Remanensen oprenses på silicagel med ethylacetat-hexan (3:1) til opnåelse af 0,126 g (96%) l,25-dihydroxy-16-dehydro-23-didehydrocholecalciferol, [a]^1 = 25 + 21,5+ (C = 0,20, MeOH).1) To a solution of 0.228 g of the product obtained in k) in 11 ml of THF is added 1.92 ml of 1M tetrabutylammonium fluoride in THF. The mixture is stirred for 16 hours. After dilution with water, the mixture is extracted with ethyl acetate. The organic phase is washed with semi-saturated brine and saturated brine, dried and evaporated. The residue is purified on silica gel with ethyl acetate-hexane (3: 1) to give 0.126 g (96%) of 1,25-dihydroxy-16-dehydro-23-didehydrocholecalciferol, [α] 20 = 25 + 21.5+ (C = 0.20, MeOH).

EKSEMPEL 2 a) På samme måde som beskrevet i eksempel lk), men ud fra 0,343 g [5S-(1Z)]-[2-[5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-2-methylencyclohexyliden] ethyl] diphenylphosphinoxid og 0,186 g af det i ek-30 sempel lj) vundne produkt), blev vundet 0,205 g (80%) (3jS,5Z,7E) ,3- [[(1,1-dimethylethyl)dimethylsilyl]oxy]-25-[(trimethylsilyl)oxy] -9,-10-secocholesta-5,7,10(19),16-tetraen-23-yn, MS m/e 580 (M+).Example 2 a) In the same manner as described in Example 1k), but from 0.343 g of [5S- (1Z)] - [2- [5 - [[(1,1-dimethylethyl) dimethylsilyl] oxy] -2-methylene cyclohexylidene ] ethyl] diphenylphosphine oxide and 0.186 g of the product obtained in Example 1j) were obtained 0.205 g (80%) (3S, 5Z, 7E), 3- [[(1,1-dimethylethyl) dimethylsilyl] oxy ] -25 - [(trimethylsilyl) oxy] -9, -10-secocholesta-5,7,10 (19), 16-tetraene-23-in, MS m / e 580 (M +).

22 DK 169945 B1 b) Ved behandling af 0,248 g af det i a) vundne produkt som beskrevet i eksempel 11) vandtes 0,153 g (91%) 25-hydroxy-l6-dehydro-23-didehy-drocholecalciferol, [a]^1 = +99,6° (c = 0,25, MeOH).B) In treating 0.248 g of the product obtained as described in Example 11) 0.153 g (91%) of 25-hydroxy-16-dehydro-23-didehydrocholecalciferol, [a] + 99.6 ° (c = 0.25, MeOH).

* EKSEMPEL 3 5 a) Til en blanding af 0,146 g lithiumaluminiumhydrid, 0,211 g natri-ummethoxid og 6,5 ml THF ved 0°C tilsættes dråbevis en opløsning af 0,180 g af det i eksempel lh) vundne produkt i 13 ml THF. Blandingen opvarmes ved 68°C i 16 timer og afkøles atter ved 0eC. Efter fortynding med 13 ml ether og tilsætning af 0,30 ml vand og 0,26 ml 10% 10 vandig NaOH omrøres blandingen ved stuetemperatur i 1 time og filtreres. De faste stoffer tritureres med ether og filtreres. De samlede filtrater inddampes og kromatograferes på silicagel med ethyla-cetat-hexan (1:2) til opnåelse af 0,179 g (99%) [1 (R*) ,1 (3E) ,3a/S,4a, -7aa)]-(3a,4,5,6,7,7a-hexahydro-l-(5-hydroxy-l,5-dimethyl-3-hexenyl)-15 7a-methyl-lH-inden-4-ol, [a]^1 = +11,5° (c = 0,33, CHC13).EXAMPLE 3 a) To a mixture of 0.146 g of lithium aluminum hydride, 0.211 g of sodium methoxide and 6.5 ml of THF at 0 ° C is added dropwise a solution of 0.180 g of the product obtained in Example 1h in 13 ml of THF. The mixture is heated at 68 ° C for 16 hours and cooled again at 0 ° C. After diluting with 13 ml of ether and adding 0.30 ml of water and 0.26 ml of 10% aqueous NaOH, the mixture is stirred at room temperature for 1 hour and filtered. The solids are triturated with ether and filtered. The combined filtrates are evaporated and chromatographed on silica gel with ethyl acetate-hexane (1: 2) to give 0.179 g (99%) [1 (R *), 1 (3E), 3a / S, 4a, -7aa]] - (3a, 4,5,6,7,7a-hexahydro-1- (5-hydroxy-1,5-dimethyl-3-hexenyl) -7a-methyl-1H-inden-4-ol, [a] + = 11.5 ° (c = 0.33, CHCl 3).

b) Til en opløsning af 0,120 g af det i a) vundne produkt i 10 ml methylenchlorid tilsættes 0,500 g pyridiniumdichromat og 25 mg pyri-dinium-p-toluensulfonat. Blandingen omrøres i 135 minutter. Efter tilsætning af 40 ml ether omrøres blandingen i 5 minutter og fil- 20 treres. De faste stoffer tritureres med ether og filtreres. De samlede filtrater vaskes med mættet vandig CuS04, vand, halvmættet vandig NaHC03 og mættet saltvand. Den organiske fase tørres og inddampes. Remanensen kromatograferes på silicagel med 35% ethylacetat-hexan til opnåelse af 90 mg (76%) [1 (R*), 1 (3E), (3a/3, 7aa)]-3,3a,-25 5,6,7,7a-hexahydro-l- (5-hydroxy-l,5-dimethyl-3-hexenyl) -7a-methyl-4H- inden-4-on, [a]g5 +30,6° (c = 0,17, CHC13).b) To a solution of 0.120 g of the product obtained in (a) in 10 ml of methylene chloride is added 0.500 g of pyridinium dichromate and 25 mg of pyridinium p-toluenesulfonate. The mixture is stirred for 135 minutes. After adding 40 ml of ether, the mixture is stirred for 5 minutes and filtered. The solids are triturated with ether and filtered. The combined filtrates are washed with saturated aqueous CuSO4, water, semi-saturated aqueous NaHCO3 and saturated saline. The organic phase is dried and evaporated. The residue is chromatographed on silica gel with 35% ethyl acetate-hexane to give 90 mg (76%) [1 (R *), 1 (3E), (3a / 3, 7aa)] - 3.3a, -25 5.6, 7,7a-hexahydro-1- (5-hydroxy-1,5-dimethyl-3-hexenyl) -7a-methyl-4H-inden-4-one, [a] g5 + 30.6 ° (c = 0, 17, CHCl3).

c) Ved behandling af 0,099 g af det i b) vundne produkt som beskrevet i eksempel lj) fås 0,111 g (89%) [1 (R*), 1 (3E), (3a/3,7aa) ]-3,3a,5,6,7, -7a-hexahydro-l-(1,5-dimethyl-5-[(trimethylsilyl)oxy]-3-hexenyl)-7a- 30 methyl-4H-inden-4-on, [α]β4 = +26,4° (c = 0,22, CHC13). t d) På samme måde som beskrevet i eksempel lk) fås ud fra 0,265 g [3S-(1Z, 3a,5/3) ] - [2- [3,5-bis [ [ (1,1-dimethyl) dimethylsilyl] oxy] -2-methy- 23 DK 169945 B1 lencyclohexyliden]ethyl]diphenylphosphinoxid og 0,095 g af det i c) vundne produkt 0,162 g (83%) (1/9,3α,5Z,7E,23E)-1,3-bis[ [ (1,1-dime-thylethyl) dimethylsilyl] oxy] -25- [ (trimethylsilyl) oxy] -9,10-secocho-lesta-5,7,10(19),16,23-pentaen, MS m/e 712 (M+) .c) By treating 0.099 g of the ib) product obtained as described in Example 1j) 0.111 g (89%) [1 (R *), 1 (3E), (3a / 3.7aa)] -3.3a are obtained. , 5,6,7, -7a-hexahydro-1- (1,5-dimethyl-5 - [(trimethylsilyl) oxy] -3-hexenyl) -7a-methyl-4H-inden-4-one, [α ] β4 = + 26.4 ° (c = 0.22, CHCl3). td) in the same manner as described in Example 1k) is obtained from 0.265 g of [3S- (1Z, 3a, 5/3)] - [2- [3,5-bis [[(1,1-dimethyl) dimethylsilyl] oxy] -2-methyl [1,2-cyclohexylidene] ethyl] diphenylphosphine oxide and 0.095 g of the ic) obtained 0.162 g (83%) (1 / 9.3α, 5Z, 7E, 23E) -1,3-bis [[(1,1-dimethylethyl) dimethylsilyl] oxy] -25- [(trimethylsilyl) oxy] -9,10-secocho-lesta-5,7,10 (19), 16,23-pentaene, MS m / e 712 (M +).

5 e) Ved behandling af 0,159 g af det i d) vundne produkt på samme måde som beskrevet i eksempel 11) fås 0,077 g (84) l,25-dihydroxy-16,23E-bisdehydrocholecalciferol, [a]j§4 = +46,5° (c = 0,20, MeOH).E) By treating 0.159 g of the product obtained in the same manner as described in Example 11), 0.077 g of (84) 1,25-dihydroxy-16,23E-bisdehydrocholecalciferol is obtained, [a] j§4 = +46 , 5 ° (c = 0.20, MeOH).

EKSEMPEL 4 a) På samme måde som beskrevet i eksempel lk) fås ud fra 0,225 g [5S-10 (1Z)]- [2- [5- [[ (1,1-dimethylethyl)dimethylsilyl]oxy]-2-methylencyclo- hexyliden]ethyl]diphenylphosphinoxid og 0,110 g af det i eksempel 3c) vundne produkt 0,150 g (81%) (3/9,5Z, 7E,23E)-3-[ [ (1,l-dimethylethyl) -dimethylsilyl]oxy]-25-[(trimethylsilyl)oxy]-9,10-secocholesta-5,7, -10(19),16,23-pentaen, [α]β4 = +68,3° (c = 0,18, CHC13).Example 4 a) In the same manner as described in Example 1k) is obtained from 0.225 g of [5S-10 (1Z)] - [2- [5- [[(1,1-dimethylethyl) dimethylsilyl] oxy] -2-methylene cyclo - hexylidene] ethyl] diphenylphosphine oxide and 0.110 g of the product obtained in Example 3c) 0.150 g (81%) (3 / 9.5Z, 7E, 23E) -3- [[(1,1-dimethylethyl) dimethylsilyl] oxy ] -25 - [(trimethylsilyl) oxy] -9,10-secocholesta-5,7, -10 (19), 16,23-pentaene, [α] β4 = + 68.3 ° (c = 0.18, CHC13).

15 b) Ved behandling af 0,144 g af det i a) vundne produkt på samme måde som beskrevet i eksempel 11) fås 0,076 g (78%) 25-hydroxy-16,23E-bisdehydrocholecalciferol, = +62,5° (c = 0,20, MeOH) .B) By treating 0.144 g of the product obtained in the same manner as described in Example 11) 0.076 g (78%) of 25-hydroxy-16,23E-bisdehydrocholecalciferol, = + 62.5 ° (c = 0) is obtained. , 20, MeOH).

EKSEMPEL 5 a) Til en opløsning af 6,25 g ethyl-3-brompropionat i 28 ml THF ved 20 -20°C tilsættes 28,8 ml 2,8M methylmagnesiumbromid i ether. Blandin gen omrøres ved stuetemperatur i 170 minutter. Efter tilsætning af 15 ml mættet vandig ammoniumchlorid og 42 ml IN HC1 skilles den organiske fase fra, og den vandige fase ekstraheres med ether. De organiske ekstrakter vaskes med mættet saltvand, tørres og inddampes.Example 5 a) To a solution of 6.25 g of ethyl 3-bromopropionate in 28 ml of THF at 20 -20 ° C is added 28.8 ml of 2.8M methylmagnesium bromide in ether. The mixture is stirred at room temperature for 170 minutes. After the addition of 15 ml of saturated aqueous ammonium chloride and 42 ml of 1N HCl, the organic phase is separated and the aqueous phase is extracted with ether. The organic extracts are washed with saturated brine, dried and evaporated.

25 Remanensen kromatograferes på silicagel med 30% ethylacetat-hexan til opnåelse af 2,57 g (45%) 4-brom-2-methyl-2-butanol, MS m/e 151 (M+-CH3).The residue is chromatographed on silica gel with 30% ethyl acetate-hexane to give 2.57 g (45%) of 4-bromo-2-methyl-2-butanol, MS m / e 151 (M + -CH 3).

b) Til en opløsning af 2,56 g 4-brom-2-methyl-2-butanol og 4,86 g imidazol i 15 ml Ν,Ν-dimethylformamid ved 0°C tilsættes 6,48 g chlor- 24 DK 169945 B1 triethylsilan. Blandingen omrøres ved stuetemperatur i 200 minutter.b) To a solution of 2.56 g of 4-bromo-2-methyl-2-butanol and 4.86 g of imidazole in 15 ml of Ν, Ν-dimethylformamide at 0 ° C is added 6.48 g of chlorine. triethylsilane. The mixture is stirred at room temperature for 200 minutes.

Efter tilsætning af is fortyndes blandingen med vand og ekstraheres med pentan. Den organiske fase vaskes med vand og mættet saltvand, tørres og inddampes. Remanensen kromatograferes på silicagel med 5 pentan til opnåelse af 4,02 g (93%) (3-brom-l,l-dimethylpropoxy) -triethylsilan, MS m/e 265 (M+-CH3) .After the addition of ice, the mixture is diluted with water and extracted with pentane. The organic phase is washed with water and saturated brine, dried and evaporated. The residue is chromatographed on silica gel with 5 pentane to give 4.02 g (93%) of (3-bromo-1,1-dimethylpropoxy) triethylsilane, MS m / e 265 (M + -CH 3).

c) Til en opløsning af 0,930 g [1 (R*) ,3aR* (3ajØ,4a,7aa) ]-3a,4,5,6,7, -7 a-hexahydro-4-hydroxy-β,7a-dimethyl-3H-inden-1-ethanol-4-methyl-benzensulfonat og 1,10 g imidazol i 73 ml methylenchlorid ved 0°Cc) To a solution of 0.930 g of [1 (R *), 3aR * (3aajØ, 4a, 7aa)] -3a, 4,5,6,7, -7 α-hexahydro-4-hydroxy-β, 7α dimethyl 3H-indene-1-ethanol-4-methylbenzenesulfonate and 1.10 g of imidazole in 73 ml of methylene chloride at 0 ° C

10 tilsættes 0,580 g chlortriethylsilan. Blandingen omrøres ved stuetemperatur i 1,5 timer. Efter tilsætning af is fortyndes blandingen med vand og omrøres i 20 minutter.0.580 g of chlorotriethylsilane is added. The mixture is stirred at room temperature for 1.5 hours. After the addition of ice, the mixture is diluted with water and stirred for 20 minutes.

Den organiske fase ekstraheres med methylenchlorid. Ekstrakterne vaskes med vand, IN N2S04, mættet vandig NaHC03 og mættet saltvand.The organic phase is extracted with methylene chloride. The extracts are washed with water, 1N N 2 SO 4, saturated aqueous NaHCO 3 and saturated brine.

15 Efter tørring og inddampning oprenses remanensen på silicagel med ethylacetat-hexan (1:5) til opnåelse af 1,22 g (100%) [l(R*),3aR*-(3a/), 4a, 7aa) ] -3a, 4,5,6,7,7a-hexahydro-4- [ (triethylsilyl) oxy] -)3,7a-dimethyl-3H-inden-l-ethanol-4-methylbenzensulfonat, [α]= +46,1° (c = 0,31, CHC13).After drying and evaporation, the residue is purified on silica gel with ethyl acetate-hexane (1: 5) to give 1.22 g (100%) [1 (R *), 3aR * - (3a /), 4a, 7aa)] - 3a, 4,5,6,7,7a-hexahydro-4- [(triethylsilyl) oxy] -) 3,7a-dimethyl-3H-indene-1-ethanol-4-methylbenzenesulfonate, [α] = +46.1 ° (c = 0.31, CHCl 3).

20 d) Til en opløsning af 3,08 g (3-brom-1,1-dimethylpropoxy)triethylsilan i 31 ml THF tilsættes 0,282 g magnesium. Blandingen opvarmes ved 68eC i 3,5 timer. Derefter omrøres en blanding af 0,686 g cuproiodid og ovennævnte Grignard-opløsning ved 3°C i 30 minutter. Til dette sættes en opløsning af 1,02 g af det i c) vundne produkt, og blandin- 25 gen omrøres ved stuetemperatur i 40 minutter. Efter tilsætning af en blanding af is og vand ekstraheres blandingen med ether. Den organiske fase vaskes med IN H2S04 og mættet vandig NaHC03, tørres og inddampes. Remanensen kromatograferes på silicagel med ethylacetat-hexan (1:15) til opnåelse af 1,80 g [1(R*),3aR*-(3a^,4a,7aa)]-3a,4,- « 30 5,6,7,7a-hexahydro-l-[1,5-dimethyl-5-[(triethylsilyl)-oxy]hexyl]-4-[(triethylsilyl)oxy]-7a-methyl-3H-inden, MS m/e 479 (M+-Et).D) To a solution of 3.08 g (3-bromo-1,1-dimethylpropoxy) triethylsilane in 31 ml of THF is added 0.282 g of magnesium. The mixture is heated at 68 ° C for 3.5 hours. Then, a mixture of 0.686 g of cuprous iodide and the above Grignard solution is stirred at 3 ° C for 30 minutes. To this is added a solution of 1.02 g of the product obtained in (c) and the mixture is stirred at room temperature for 40 minutes. After adding a mixture of ice and water, the mixture is extracted with ether. The organic phase is washed with 1N H2SO4 and saturated aqueous NaHCO3, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:15) to give 1.80 g [1 (R *), 3aR * - (3a ^, 4a, 7aa)] - 3a, 4, - 7,7a-Hexahydro-1- [1,5-dimethyl-5 - [(triethylsilyl) oxy] hexyl] -4 - [(triethylsilyl) oxy] -7a-methyl-3H-indene, MS m / e 479 (M + -Et).

e) Til en opløsning af 1,60 g af det i d) vundne produkt i 5 ml THF tilsættes 2,00 ml 1M tetrabutylammoniumfluorid i THF. Blandingen opvarmes ved 68°C i 50 minutter. Efter afkøling til stuetemperatur 25 DK 169945 B1 fortyndes blandingen med vand og ekstraheres med methylenchlorid. Den organiske fase vaskes med saltvand, tørres og inddampes. Remanensen oprenses på silicagel med ethylacetat-hexan (1:1) til opnåelse af 0,420 g (79%) [1(R*) ,3aR*-(3a/3,4aa, 7aa) ]-3a,4,5,6,7,7a-hexahydro- 5 4-hydroxy-α-α-e,7a-tetramethyl-lH-inden-l-pentanol, [αΐβ1 = +12,0° (c = 0,25, CHCI3).e) To a solution of 1.60 g of the product obtained in d) in 5 ml of THF is added 2.00 ml of 1M tetrabutylammonium fluoride in THF. The mixture is heated at 68 ° C for 50 minutes. After cooling to room temperature, the mixture is diluted with water and extracted with methylene chloride. The organic phase is washed with brine, dried and evaporated. The residue is purified on silica gel with ethyl acetate-hexane (1: 1) to give 0.420 g (79%) [1 (R *), 3aR * - (3a / 3.4aa, 7aa)] -3a, 4.5.6 , 7,7a-Hexahydro-4-hydroxy-α-α-e, 7α-tetramethyl-1H-indene-1-pentanol, [αΐβ1 = + 12.0 ° (c = 0.25, CHCl 3).

f) Til en opløsning af 0,210 g af det i e) vundne produkt i 18 ml methylenchlorid tilsættes 0,870 g pyridiniumdichromat og 44 mg pyri-dinium-p-toluensulfonat. Blandingen omrøres i 175 minutter. Efter 10 tilsætning af 50 ml ether omrøres blandingen i 5 minutter og filtreres. De faste stoffer vaskes med mættet vandig CuS04, vand, halv-mættet vandig NaHC03 og mættet saltvand. Den organiske fase tørres og inddampes. Remanensen kromatograferes på silicagel med 35% ethylacetat-hexan til opnåelse af 0,175 g (84%) [1 (R*) ,3aR*- (3a^S, 7aa) ] -3,3a, -15 5,6,7,7a-hexahydro-l-(5-hydroxy-1,5-dimethylhexyl)-7a-methyl-4H- inden-4-on, [ajp1 = +28,2° (c = 0,22, CHCI3).f) To a solution of 0.210 g of the product obtained in e) in 18 ml of methylene chloride is added 0.870 g of pyridinium dichromate and 44 mg of pyridinium p-toluenesulfonate. The mixture is stirred for 175 minutes. After adding 10 ml of ether, the mixture is stirred for 5 minutes and filtered. The solids are washed with saturated aqueous CuSO4, water, semi-saturated aqueous NaHCO3 and saturated saline. The organic phase is dried and evaporated. The residue is chromatographed on silica gel with 35% ethyl acetate-hexane to give 0.175 g (84%) [1 (R *), 3aR * - (3a ^ S, 7aa)] -3.3a, -15 5.6.7, 7α-Hexahydro-1- (5-hydroxy-1,5-dimethylhexyl) -7α-methyl-4H-inden-4-one, [α] β1 = + 28.2 ° (c = 0.22, CHCl 3).

g) Ved behandling af 0,168 g af det i f) vundne produkt på samme måde som beskrevet i eksempel lj) fås 0,211 g (100%) [1 (R*) ,3aR*-(3ajØ,-7aa)]-3,3a,5,6,7,7a-hexahydro-l-(l,5-dimethyl-5-[(trimethylsilyl)- 20 oxy]hexyl)-7a-methyl-4H-inden-4-on, [α]β® * +21,9° (c = 0,27, CHCI3).g) By treating 0.168 g of the obtained product in the same manner as described in Example 1j), 0.211 g (100%) of [1 (R *), 3aR * - (3aJ0, -7aa)] - 3.3a is obtained. 5,6,7,7a-hexahydro-1- (1,5-dimethyl-5 - [(trimethylsilyl) oxy] hexyl) -7a-methyl-4H-inden-4-one, [α] β® + 21.9 ° (c = 0.27, CHCl 3).

h) På samme måde som beskrevet i eksempel lk) fås ud fra 0,581 g [3S-(1Ζ,3α,5/9) ] - [2- [3,5-bis[[(l,l-dimethyl)dimethylsilyl]oxy] -2-methylen-cyclohexyliden] ethyl] diphenylphosphinoxid og 0,210 g af det i g) vundne produkt 0,358 g (83%) (1α,3/?,5Ζ,7E)-l,3-bis[[(1,1-dimethyl- 25 ethyl)dimethylsilyl]oxy]-25-[(trimethylsilyl)oxy]-9,10-secocholesta- 5,7,10(19),16-tetraen, MS m/e 714 (M+) .h) In the same manner as described in Example 1k) is obtained from 0.581 g of [3S- (1Ζ, 3α, 5/9)] - [2- [3,5-bis [[(1,1-dimethyl) dimethylsilyl] oxy] -2-methylene-cyclohexylidene] ethyl] diphenylphosphine oxide and 0.210 g of the product obtained 0.358 g (83%) (1α, 3β, 5Ζ, 7E) -1,3-bis [[(1,1 -dimethyl-ethyl) dimethylsilyl] oxy] -25 - [(trimethylsilyl) oxy] -9,10-secocholesta-5,7,10 (19), 16-tetraene, MS m / e 714 (M +).

Ved behandling af 0,350 g af det i h) vundne produkt som beskrevet i eksempel 11) fås 0,168 g (83%) l,25-dihydroxy-16-dehydrocholecal-ciferol, [a]fj° = +40,0° (c = 0,17, MeOH) .By treating 0.350 g of the product obtained as described in Example 11), 0.168 g (83%) of 1,25-dihydroxy-16-dehydrocholecal ciferol, [a] fj ° = + 40.0 ° (c = 0.17, MeOH).

26 DK 169945 B1 EKSEMPEL 6 a) På samme måde som beskrevet i eksempel lk) fås ud fra 0,383 g [5S-(1Z) ] -[2-[5-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-2-methylen-cyclohexyliden]ethyl]diphenylphosphinoxid og 0,188 g af det i ek- 5 sempel 5g) vundne produkt 0,245 g (78%) (3a,5Z,7E)-3-[ [(1,1-dimethyl-ethyl)dimethylsilyl]oxy]-25-[(trimethylsilyl)oxy]-9,10-secocholesta- if 5,7,10(19),16-tetraen, [α]β4 = +67,5° (c = 0,20, CHC13).Example 6 a) In the same manner as described in Example 1k), is obtained from 0.383 g of [5S- (1Z)] - [2- [5 - [[(1,1-dimethylethyl) dimethylsilyl] oxy] - 2-methylene-cyclohexylidene] ethyl] diphenylphosphine oxide and 0.188g of the product obtained in Example 5g) 0.245g (78%) (3a, 5Z, 7E) -3- [[(1,1-dimethyl-ethyl) dimethylsilyl] oxy] -25 - [(trimethylsilyl) oxy] -9,10-secocholesterol 5,7,10 (19), 16-tetraene, [α] β4 = + 67.5 ° (c = 0.20 , CHC13).

b) Behandling af 0,239 g af det i a) vundne produkt på samme måde som beskrevet i eksempel 11) giver 0,135 g (83%) 25-hydroxy-16-dehydro- 10 cholecalciferol, [<*]ρ^ = +75,4° (c = 0,13, MeOH) .b) Treatment of 0.239 g of the product obtained in the same manner as described in Example 11) gives 0.135 g (83%) of 25-hydroxy-16-dehydro-cholecalciferol, [<*] ρ ^ = +75.4 ° (c = 0.13, MeOH).

Følgende eksempler A og B eksemplificerer sammensætningen af bløde gelatinekapsler til oral administration og af en topisk creme:The following Examples A and B exemplify the composition of soft gelatin capsules for oral administration and of a topical cream:

EKSEMPEL AEXAMPLE A

mg/kapsel 15 Forbindelse E 0,0001-0,010mg / capsule Compound E 0.0001-0.010

Butyleret hydroxytoluen 0,016Butylated hydroxytoluene 0.016

Butyleret hydroxyanisol 0,016Butylated hydroxyanisole 0.016

Fraktioneret kokosnøddeolie 160,0 i 27 DK 169945 B1Fractionated coconut oil 160.0 in 27 DK 169945 B1

EKSEMPEL BEXAMPLE B

mg/g crememg / g cream

Forbindelse E 0,001-1,0Compound E 0.001-1.0

Cetylalkohol 1,5 5 Stearylalkohol 2,5Cetyl Alcohol 1.5 Stearyl Alcohol 2.5

Sorbitanmonostearat 2,0Sorbitan monostearate 2.0

Glycerylmonostearat og polyoxyethylenglycolstearat 4,0Glyceryl monostearate and polyoxyethylene glycol stearate 4.0

Polysorbat 60 1,0 10 Mineralolie 4,0Polysorbate 60 1.0 10 Mineral Oil 4.0

Propylenglycol 5,0Propylene Glycol 5.0

Propylparaben 0,05Propylparaben 0.05

Butyleret hydroxyanisol 0,05Butylated hydroxyanisole 0.05

Sorbitolopløsning 2,0 15 Edetatdinatrium 0,01Sorbitol Solution 2.0 15 Edetate Disodium 0.01

Methylparaben 0,18Methylparaben 0.18

Destilleret vand q.s. ad 100 gDistilled water q.s. ad 100 g

Claims

Compound according to Claim 1, characterized in that it is selected from: 1.25-dihydroxy-16-dehydro-23-didehydrocholecalciferol, 10 1,25-dihydroxy-15,23E-bisdehydrocholecalciferol and 1.25-dihydroxy-16-dehydrooholecalciferol.

A compound according to claim 1 or 2 for use as a therapeutically active substance, particularly for the treatment of hyperproliferative skin diseases, especially psoriasis, or for the treatment of neoplastic diseases, especially leukemia.

Process for the preparation of a compound according to claim 1 or 2, characterized in that a corresponding compound of formula I, which instead of two or three hydroxy groups contains two or more? Wherein R 1 and R 3 are C 1-4 alkyl and R 2 is -alkyl, aryl or aryl - C 1-4 alkyl, reacted with a means capable of removing the protecting groups. A drug, particularly for the treatment of hyperproliferative skin diseases, especially psoriasis, or for the treatment of neoplastic diseases, especially leukemia, comprising an effective amount of a compound of claim 1 or 2 and a pharmaceutically effective carrier, particularly for oral or topical administration.

Use of a compound according to claim 1 or 2 for the manufacture of a medicament for the treatment of hyperproliferative skin diseases, especially psoriasis, or for the treatment of neoplastic diseases, especially leukemia.