AU622139B2 - 16-dehydro-vitamin d3-derivatives - Google Patents

Description

4 S F Ref: 82646 FORM COMMONWEALTH OF AUSTRALIA PATENTS ACT 1952 COMPLETE SPECIFICATION 62 2 139

(ORIGINAL)

FOR OFFICE USE: Class Int Class fe S Complete Specification Lodged: Accepted: Published: S Priority: A Related Art: Name and Address of Applicant: F Hoffmann-La Roche Co Aktiengesellschaft Grenzacherstrasse 124-184 CH-4002 Basle

SWITZERLAND

Spruson Ferguson, Patent Attorneys Level 33 St Martins Tower, 31 Market Street Sydney, New South Wales, 2000, Australia Address for Service: Complete Specification for the invention entitled: 16-Dehydro-V'tamin 03-Derivatives The following statement is a full description of this invention, including the best method of performing it known to me/us 5845/3 -1 RAN 4212/54 Abstract The novel compounds of the formula

'A

It t t t rI ;t 11

V

wherein R is hydrogen or hydroxy and A is -CHE-C-; CI-I-CH- with E-configuration or -CH 2

CHI

2 are useful as agents for the treatment of hyperprolifer-tive disorders of the skin and as agents for the treatment of neo-plastic diseases.

£41

I

I I I t I I I I I -C C- 01

I"*

*9 94 9( 9 I t 4 tt I y o eoo t RAN 4212/54 The invention relates to compounds of the formula

'A

"1

OH

wherein R is hydrogen or hydroxy, and A is -CEC-, -CH=CH- with E-configuration or -CH 2

-CH

2 to pharmaceutical compositions comprising one, two or more compounds of formula I, and to the use of said compounds for the manufacture of such compositions useful in the treatment of hyperproliferative skin diseases, such as psoriasis, and for the treatment of neoplastic diseases, such as leukemia.

Examples of C -alkyl groups as referred to below, are methyl, ethyl, propyl, isopropyl, butyl and t-butyl.

Examples of aryl-C -4-alkyl groups are benzyl, phenethyl 30 and phenylpropyl. Examples of aryl groups are phenyl and p-tolyl. Halogen denotes bromine, chlorine, fluorine or iodine.

Compounds of formula I of the invention are compounds A *4*4c t1 0 t 9 9- M6/14.12.88 I A t I -2 to F as defined below: 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-dehyro-23--didehydrocholecalciferol; and F: 25-hydroxy-16-dehydro-23-didehydrocholecalciferol; among which the 1,25-dihydroxylated compo',ids A, C and E are preferred.

The compounds of formulae la and Ib (encompassed by formula I) can be prepared as described in the Schemes 1, 2 j 5 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 of the formula -OSi(R 1

,R

2

R)

2, 3 wherein Rand R 3 are C 1 4 -alkyl and Ris 1 3 1-42 C 1-alkyl. aryl or aryl-C 1-alkyl, with an agent capable of removing the protecting groups.

CC 77 -3 Scheme 1 CS R CS-'1 2

H]

49*4 4 .4.4 44 4 4 44., 09 4 9 4. 4 4, 4 14 4£ 44 t 1~ 4 4 4 4£ ttt~c 4 4444 4.

.4 0 444444 4 4 S8I, p-I-.

83 8, 0-Si-8 2 82

A.

1 wherein A is as described above and R 1 and R 3 are independently C 1-alkyl and R 2is independently C 1-alkyl, aryl, or aryl-C 1-alkyl.

H

-4 Scheme 2 *999 a 9** 9~ 49*a 9* 9 9 99.9 99 9 9 *9 9. 9 9 9* 99 p 9~ *a a a* 9 9 99*944 a 9999 a a 44 a 9949 #4 9 4 'OS 2 wherein RV, R 2 and R 3 are as described above.

[F

Scheme 3 OSI as Ix S3C *64 4 *44* .4,4 4494 4* 9 4 4..

99 4 0 .4 .4 S 4 44 4S 4,44 P 44 4. t .4 .4 9 94 P44444 4 4 44 *4 4 4.

44 4 4 w~44 49 O 9 wherein RV, R 2 and R3are as described above, X is chlorine, bromine or iodine and Ts is tosyl.

6 The intermediates of formula II, encompassing those of formulae IIa, lib and lic, are novel and are part of the invention.

In Scheme 1. the compound of formula II is converted to a compound of formula IVa or IVb by reaction with the corresponding compound of formula -pOPh 2 I r CT

RI

R2 S iv, R4

R

3 where Ph is phenyl; R 4 is H or -OSi(Rlo R 2 R 3).

and R 1

R

2 and R 3 are as described above.

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 S, a strong base, such as an alkyl lithium, e.g. butyl lithium.

The protecting groups of a compound of formula IVa or IVb are removed by reaction with a fluorine calt, e.g, tetrabutylammonium fluoride in a polar, organic solvent, e.g. ether or preferably THF, to yield a corresponding compound of formula la or Ib.

In Scheme 2, the compound of formula V is oxidized to the compound of formula VI by treatment with an oxidizing agent, e.g. 2,2'-bipyridinium chlorochromate or preferably, pyridinium chlorochromate, in an aprotic, organic solvent such as methylene chloride.

7 The compound of formula VI is converted to a compound of formula TIb, by reaction with, a (trialkylsilyi)imidazole such as (trimethylsilyl)imidazole, in an aprotic organic solvent such as THF, or preferably, methylene chloride.

The compound of formula V may also be partially hydrogenated to the compound of formula VII by reaction with a reducing agent, e.g. lithium aluminium hydride, preferably in the presence of an alkali metal alkoxide, e.g. sodium methoxide, in an aprotic organic solvent e.g. ether, or preferably THF, at reflux temperature (about 68 0 C for THF), for about 10-20 hours.

The resulting compound of formula VII is oxidized to the S* compound of formula VIII by treatment with an oxidizing agent as described above for the oxidation of V to VI, S' The compound of formula VIII is converted to a compound S, 20 of formula IIa, by reaction with a (trialkylsilyi)imidazole.

as described above for the conversion of VI to 1Ib.

In Scheme 3, the compound of formula X is reacted in ether, preferably THF, at reflux temperature with magnesium.

The resulting Grignard solution is treated with cuprous iodide and then the compound of formula IX is added.

The resulting compound of formula XI is reacted with a fluoride salt, e.g. tetrabutylammonium fluoride in ether or preferably THF.

The obtained compound of formula XII may be oxidized as described above for the oxidation of V to VI.

2 8 The resulting compound of formula XIII is converted to a compound of formula IIc, by reaction with a (trialkylsilyl)imidazole as described above for the conversion of VI to IIb.

To prepare a compound of formula IX, the compound of formula

XIV

I,

v str Ie I 1 1 I St 15 (Tetrahedron 40, 1984,2283) can be 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 formula SXV xv HO H is then converted to a compound of formula IX by reaction of a trialkylsilyl chloride, e.g. trimethylsilyl chloride, in the presence of imidazolo and in an aprotic organic solvent, e.g. THF or methylene chloride.

To prepare a compound of formula X, a compound of formula j t X-CHl 2

CH

2 COCH 3

XVI

<r I 7 9 can be converted to a compound of formula

X-CH

2 CH 2

C(CE

3 )2-1 XVII wherein X is as above, by reaction with a methyl Grignard reagent such as methylmagnesium bromide in ether. The compound of formula XVII is cotnverted to a compound of formula X, by reaction with a trialkylsilyl chloride, as described above for converting XV to IX.

For preparing 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 15 100 0 C for 1 to 5 hours to give a compound of formula

'I.

*1 t It ~t tt t $t Ir I I *4

*T]

XVIII

This is converted to the compound of formula

XIX

by reaction with a reducing agent, e.g. ditsobutylaluminium hydride, followed by hydrolysis with, e.g. a mineral acid, ,i 10 such as hydrochloric acid. The reduction is conducted in an aprotic organic solvent, e.g. methylene chloride, at about to 10 0 C for about 20 to 90 minutes. The compound of formula XIX is converted to the compound of formula Br Br *r 4 I *I 4 by reaction with a mixture of triphenylphosphine, carbon tetrabromide and zinc dust, in an aprotic organic solvent, e.g. methylene chloride, for about 1 to 30 hours.

The compound of formula XX is converted to the compound of formula o t r Sti it.

4 O r I 4 44

J

XXI

ts' Ie I by reaction with a strong base, e.g. butyllithium, in a polar aprotic solvent, e.g. THF, at about -80 to -70 0 C, for about 1 to 3 hours. The compound of formula XXI is converted to the compound of formula fU- I i 11

XXII

H

Me3SiO by reaction with (trimethylsilyl)imidazole in an aprotic organic solvent, e.g. THF or methylene chloride. This compound is converted to the compound of formula set, OH XXIII Me3SiO ee 2 by reaction with a strong base, e.g. butyllithium and then with acetone. The reaction is conducted in an aprotic organic solvent, e.g. THF at about -80 to -60 0 C. The 25 compound of formula XXIII is deprotected to give the compound of formula V (in Scheme 2) by reaction with a fluorine salt, e.g. tetrabutylammonium fluoride in an organic solvent, e.g. ether or THF.

rrc4ol The compound of formula I stimulate differentiation and decrease proliferation of human keratinocytes. Accordingly, they are useful as agents in the treatment of hyperproliferative skin disorders, such as psoriasis, basal cell carcinomas, disorders or keratinization and keratosis. The compound of formula I are also useful as agents in the treatment of neoplastic diseases, such as leukemia.

12 The activity of compounds of formula I as agents for the treatment of hyperproliferative skin diseases can be demonstrated e.g. by test procedures known in the art, such as set forth in The Society for Investigative Dermatology 1986, 709-714. The effects of the compounds A to F above on the morphologic differentiation of cultured human keratinocytes compared to the effect of 1,25-dihydroxycholecalciferol (compound X) are expressed in the Tables 1 4 to 4 below, as number (xlO of human keratinocytes in culture. A compound which induces the differentiation of basal cells to squamous and envelope cells is useful as an agent in the treatment of skin diseases characterized by disorders of keratinization, such as psoriasis.

o 15 Table 1 4 Compound Dose Number of cells (xlO

(M)

t total basal squamous envelope Control 133±5 118£4 15_1 18t2 X 10-10 122*4 103±2 19±2 2311 10-8 112±6 89±2 23±4 3013 10-6 95±7 6416 31±1 34±2 A 10-10 132±8 115±7 17±1 2742 10-8 128±10 10618 2212 33t2 10-6 101±7 71,±5 30£2 3912 B 10-10 133t6 115±5 18L1 25i1 10-8 13114 10912 22±2 2912 10-6 104±4 74±3 30±1 33t1 13

'I.

1* Table 2 Compound Dose Number of cells (x10 total basal squamous envelope Control 10-1.0 la-8 10-6 10-1.0 10-8 10-6 10-1.0 10-8 10-6 123 ±7 116±9 101±10 8 3±5 117±4 10 8 ±3 8 0±7 113 ±7 111±7 9 4 ±3 105 ±6 9518 7 5 ±8 5 7±4 92±2 8 0±2 5 4±6 9 3±6 86i±3 6 8±1 18± 1 2 1±1 2 6±2 2 6±1 2 5±2 2 8±1 2 6 ±1 20±1 2 5±2 26 ±2 74 ±7 91±4 122 ±1 1 146:L16 10316 12 8 ±3 15 3±1 1.04 1281±5 144*t7 t 9 4 s~ 9 9 1 I It *9 1 I I I Itt~ It, Table 3 Compound D or.; e 4 Number of cells (xlO total basal squamous envelope Control

X

10-1.0 10-8 10-6 10O-1.0 10-8 10-6 10-10 10-8 10-6 108 ±10 106 ±7 84 ±8 7 3±7 8 6±4 8 2± 3 78±3 103 ±5 97 ±3 8 4±4 9 3±8 8 6±6 6 1±5 5 1±5 6 3±2 5 3±2 4 1±1 8 1±3 67 ±2 5 5±2 15 ±2 18 ±1 2 3± 3 2 2±2 2 3±2 2 9±1 27 ±2 22±2 2 9± 1 2 9±1I 8 8 ±8 100± 9 12 2 ±8 14211l 114 14 14 7 4 103 ±4 12 1±6 13 7 ±7 The activity of compounds of formula I as agents for the treatment of hyperproliferative skin diseases can also be by determining the number of human keratinocytes grown and the number of envelopes formed, as well as the number squamous carcinoma cell lines (SCC-115) grown in 4i "2 14 cultures, in the presence of said compounds. The results are given in the Tables 4 and Table 4.

Treatment Dose

(M)

Number of keratinocytes (xlO 4) Number of envelopes (XIO 2) Control ethanol) 189.49±22.3 858. 28±185 Compound E 6 t r 10o 12 10-10 10-8 10-6 187.36±15.33 175.34±10.19 145.79±15.66 41.95± 7.53 1136.63± 383.66 1444.87± 312.47 2113.62±1049.33 1916.83± 887.66 Control ethanol) ,.lompound A 1 4 It I t 108 1 106 1 148.73±16.23 114.91±10.95 130-37±24.32 120.67±16.87 109.22±15.87 16 62.2± 3973.81 72 3 5.2 83 2 3.5 420. 1 126.99 55.5 157. 6 2193.71 921.9 v Ir ii Li :i n 15 Table Treatment Dose

(M)

Control Compound E 10-12 1010 10-8 io6 Number of cell lines (xlO 5 7.35±1.75 6.98±1.68 5.89±1.58 5.76±1.53 0.40±0.98 0.49±0.13 *r4e r ?r (i 4F I i Compound A 10- 6 The above results show that the compounds of formula I induce differentiation of skin cells and, accordingly, are useful in the treatment of hyperproliferative disorders of the skin, such as psoriasis.

In order to demonstrate the activity of the compounds of formula I as agents for the treatment of neoplastic diseases, the anti-proliferative (AP) and differentiation- -inducing (DI) effects of the compounds A to F and human promyelocytic HL-60 tumor cells were evaluated. In Table 6 the AP effect is given in percent reduction of cell number and in concentration ID 50 of the compound which reduced the cells number by 50%. The DI effect is expressed as the percentage of differentiated cells and as the concentration

ED

50 of the compounds which induced a 50% differentiation of the cells.

It t I 16 Table 6 Concentration (xlO- M) Reduction in cell number ID 5 0 -8 (x 10 M) Differentiated cells ED 5 0 (x10 M f t I 11 I Ct

I

I

I It Compound X 0.01 0.1 1 100 15 Compound A 0.01 0.1 1 10 100 Compound B 0.1 1 100 1000 6 5 16 66 84 10 33 84 85 85 10 8 14 82 95 2 0.2 35 3 11 19 68 98 3 16 92 97 98 4 6 93 2 0.2 32 1 T

I

17 Compound C 0.01 18 3 0.1 20 19 1 81 0.3 92 0.3 85 97 100 86 99 Compound D 0.1 12 1 1 12 2 17 150 17 200 100 46 31 4 15 1000 95 97 t Vt Compound E 0.01 6 9 0.1 59 1 80 0.07 96 0.1 10 81 98 t Compound F 0.1 13 4 1 10 12 8 70 21 100 58 1000 95 91 These data indicate that each of the compounds in question restrains the proliferation of human promyelocytic cells, in vitro, even though they are not toxic to the cells. Furthermore, the cells differentiate toward a more mature phenotype at the same doses which inhibit proliferation. From these results it can be seen that each i 18 of the compounds tested is useful as an agent in the treatment of neoplastic diseases, such as leukemia.

The compounds of formula I can be administered orally for the treatment of neoplastic diseases or for the treatment of hyperproliferative skin diseases, to warmblooded animals, which need such treatment, e.g. to an adult human, in dosage that are in the range of about 0.1 to 4g per day.

For the treatment of hyperproliferative skin diseases the compounds of formula I can also be administered topically to warmblooded animals which need such treatment, in dosage of about 1 to 1000 Ig per gram of topical 15 formulation per day.

Oral dosage forms comprising compounds of formula I may S, be incorporated e.g. in capsules or tablets with S: pharmaceutically acceptable carriers. Examples of such carrier materials which may be incorporated into capsules are binders, such as gum tragacanth or gelatin; excipients, such as dicalcium phosphate; disintegrating agents, such as corn starch; lubricants, such as magnesium stearate; sweetening agents, such as sucrose; flavoring agents, such as peppermint. Tablets may be coated with shellac, sugar or both. A syrup or elixir may contain a sweetening agent, methyl and propyl parabens as preservatives, a dye and a flavoring agent.

Topical dosage forms comprising compounds of formula I include ointments and creams encompassing formulations having oleaginous, adsorbable, water soluble and emulsion-type bases, such as lanolin and polyethylene glycols. Topical dosage forms also comprise gels, lotions, powders and aerosols. The topical compositions can also be Semployed in the treatment of inflammations of the skin or of mucous membranes, e.g. the mucous lining of the mouth or 'i 19 lower colon.

b ,I 6 a a ;6 4$ A4 Lotions, i.e. liquid preparations varying from simple solutions to aqueous of hydroalcoholic preparations containing finely divided substances, can contain suspending or dispersing agents, such as cellulose derivatives, e.g.

ethyl or methyl cellulose; gelatin o: gums, which incorporate the active ingredient in a vehicle made up of water, alcohol or glycerin. Gels are semi-solid preparations made by gelling a solution or suspension of the active ingredient in a carrier vehicle. The vehicles, which can be hydrous or anhydrous, are gelled using a gelling agent, e.g.

carboxy polymethylene, and neutralized to a proper gel consistency with the use of alkalies, e.g. sodium hydroxide, or amines, such as polyethylenecocoamine.

Example 1 a) A mixture of 3.24 g of [1(R*),3aR*-(3a8,4a,7ac)]- 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 is stirred at 00 for 18 hr. After addition of ice and dilution with water, the mixture is extracted with methylene chloride. The organic phase is washed with IN 25 H 2

SO

4 suaturated NaHCO 3 then dried and evaporated.

The residue is chromatographed on silica gel with ethyl acetate-hexane to afford 4.61 g of [1(R*),3aR*-(3a8,4a,7aa)]-3a,4,5,6,7,7a-hexahydro- 4-hydroxy-3,7a-dimethyl-3H-indene-1-ethanol -4-methyl- 21 benzenesulfonate [a]D 31.90 (c 0.53, CHC13).

b) To a solution of 4.61 g of the product of a) in 22 tal 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 pumped to remove the solvent, then diluted with water. The mixture is extracted with ether. The organic phase is washed with saturated brine, r*- 20 dried and evaporated. The residue is chromatographed on silica gel with methylene chloride-hexane-ethyl acetate (86:7:7) to give 2.52 g of (3aB,4a,7aa)]-3a,4,5,6,7,7a-hexahydro-4-hydroxy-3,7a- 21 dimethyl-3H-indene-l-propanenitrile, Ca]D 29.20 (c 0.65, CHC13).

c) To a mixture of 6.85 ml of diisobutylaluminium hydride in hexane and 5.2 ml of methylene chloride at -6 0 C is added a solution of 0.430 g of the product of b) in 10 ml of methylene chloride. The mixture is stirred at -6°C for minutes. After addition of saturated ammonium chloride, the mixture is hydrolyzed with 3N HCl-ether The aqueous layer is extracted with ether. The organic layers are washed 5 with saturated brine, dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane to afford 260 mg 1[(R*),3aR*-(3a,4a,7aa)]iot -3a,4,5,6,7,7a-hexahydro-4-hydroxy-B,7a-dimethyl-3H-indener 22 -1-propanal, 43.10 (c 0.32, CHC13) d) A mixture of 1.77 g of triphenylphosphine, 2.23 g of I, carbon tetrabromide, 441 mg of Zn dust and 23 ml of Smethylene chloride is stirred at 25 0 C for 31 hours. To this mixture is added a solution of 0.430 g of the product of c) in 38 ml of methylene chloride and the mixture is stirred for 18 hours. The mixture is diluted with pentane and insoluble material is filtered off. The insoluble fraction is dissolved in methylene chloride and the solution is again diluted with pentane. After filtration, the combined filtrates are evaporated. The residue is purified on silica gel with 1:4 ethyl acetate-hexane to give 0.490 g of [l(R*),3aR*-(3a,4I,7aa)]-l-(4,4-dibromo- 1-methyl -3-butenyl)-3a,4,5,6,7,7a-hexahydro-7a-methyl-3H- 22 indene-4-ol, [a]D 14.4 (c 0.55, CHCI 3 e) To a solution of 0.680 g of the product of d) in 31 ml of THF at -75 0 C are added dropwlse 3.77 ml of 1.6M solution 21 of butyllithium in hexane. The mixture is stirred at -75 0

C

for 1 hour and at 25 0 C for 1 hr. After addition of saturated brine, the mixture is diluted with saturated aqueous NabCO3 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 to afford 0.350 g of (3a3,4a,7a)]-3a,4,5,6,7,7a-hexahydro -7a-methyl-l- 22 (1-methyl-3-butynyl)-3H-inden-4-ol, CjD 30,'/0 (c 0.42, CHC1 3 f) To a solution of 1.29 g of the product of e) in 80 ml of methylene chloride are added 3.59 g of 1-(trimethylsilyl)imidazole. The mixture is stirred at 25 0 C for 3 hours. After 15 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 C. to: acetate-hexane (1:15) to give 1.70 g of (3a3,4a,7aa)-3a,45,6,7,7a-hexahydro -7a-methy-l- (1-methyl-3-butynyl)- 4-[(trimethylsilyl)oxy -3H-indene, aJD 39.70 (c 0.30, CIC1 3 C

C.

g) To a solution of 1.70 g of the product of f) in 48 ml of 25 THEF at -750C are added dropwise 6.01 ml of 1.6M butyllithium in hexane. After stirring for 40 minutes, 3,05 ml of acetone are added and the mixture is stirred at -75 0 C for 20 minutes i*Ce and at 25 0 C for 75 minutes. After addition of 40 ml of a 1:1 mixture of 2M KHCO and IM potassium sodium tartrate, the 3 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 to give 1.62 g of C1(R*),3aR*(3aj3,4cL,7am)).6-(3a, 4,S,6,7,7a-hexahydro-7a-mehyl-4.[(timethylsilyl)oxyj- 20 3H-indon-1-yl)-2-methvl-3-heptyn-1-ot, ta) 39.70 -22- (c 0.30, Cl-I 3 h) To a solution of 1.62 g of the product of g) in 53 ml of THE' are added 15.5 ml of IM tetrabutylammonium fluoride in THE'. The mixture is stirred for 50 minutes. After dilution with half saturated NaH-CO 0 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 chromatogcaphed on silica gel with ethyl acetate-hexane to give 1.17 g of -1-(1,5-di-mohyl-5-hydroxy-3-hexynyl)-7a-methyl-3-inden--ol, m.p. 105-1070.

i)To a solution of 0.720 g of the product of h) in 44 ml of methylene chloride are added 1.59 g of sodium acetate and 3.18 g of 2,2'-bipyridinium chlorochromate, The mixture is stirred for 2 hours. Additional 1.59 g of 2,21-bipyridinium chlorochromate are then added and the stirring continued for 2 hours. Then, after the addition of 6 ml of 2-propanol, the mixture is diluted with water and extracted with ether-.othyl acetate The organic phase Is washed with water, IN H 2 soV saturated NaHCO 3 and saturated brine. After drying, the solution Is evaporated and the residue chromatographed on silica gel with ethyl acetate-hexane to give 0.560 g of [l(R*),3aR*-(3aI3,7aa)]-.

3a 7a-hexahydro-l-(5-hydroxy-1, 20 -3-hexynyl)-7a-mthyl-4l-i-nden--4-one, (o'j D +35.30 (c O6, CHCl 3 303 j)To a solution of OqsZ g of the product of I) in 70 ml of methylene chloride are added 2.00 g of 1-(trimethylsilyl)imidazole. After stirring for 17 hours and addition of 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 Ise chrornatographed on silica gel with ethyl acetate.iiexanlo to give 0.693 g

I

I 23 of [1(R*),3aR*-(3a3,7aa)1-33a,5,6,7,7a-hexahydro -1-(1,5-dimethyl-5-[(trimethylsilyl)oxy]-3-hexyfyl) 20 -7a-methyl-41-lnden-4-oe, 29.50 (c 0.20,

D

CHCl 3 k) To a solution of 2.00 g of C3S-(IZ,3ot,513)]- (2-E3.5-bisE E (2,1-dimethyl)dimethylsilyljoxyj-2-methylenecyclohexylidenejethyl]diphenylphosphine oxide in 45 ml of THF at -75 0 C are added dropwise 1.87 ml of 1.6M butyllithiom in hexane. After stirring for 6 minutes a solution of 0.693 g of the product of j) in 26 ml of THE are added dropwise. After stirring at -75 0 C for 70 minutes and addition of a 1:1 mixture of IM potassium sodium tartrate and 2M KHCO 3 the mixture Is extracted with ethyl acetate.

*43* 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 of (lo (I,3j3,5,7E)-L1,-is(Id-imethletyldemthlsyy -oxy-25-((trimethylsilyl)oxyj-9,0-secocholesta-5,7,10(19),16- 23 tetraene-3-yne, CaJD 47.10 (c 0.21, CHM 3 1) To a solution of 0.228 g of the product of k) In 11 ml of TF are added 1.92 ml of 1M tetrabutylammonium fluoride in TFIF. The mixture is stirred for 16 hours. After dilution with water, the mixture Is extracted with ethyl acetate. The organic phase Is washed with half saturated brine and saturated brine, dried and evaporated. The residue is purified on silica gel with ethyl acetate-hexane to afford 0.126 g of 1,25-dihydroxy-16-dehydro-23.didehydrocholecalciferol, Cal 21 +21.5+ (c 0.20, MeOHl).

[D

a) As describod In Example lk), but starting from 0.343 g of tS5S- (1Z) 4 S uthlc l~yI)dieti try lfoxyj -2-methylo ecyclohoxylidenejethyli dtphonylphosphino oxide and 0.186 q of the product of Examplo thera wore F M N W-- 41 -1 24 obtained 0.205 g of (3,5Z,7E)-3-[[(1,1-dimethyl- -9,10-secocholesta-5,7,10(19),16- tetraen-23-yne, MS m/e 580 b) By treating 0.248 g of the product of a) as described in Example II), there were obtained 0.153 g of 25-hydroxy-16-dehydro-23-didehydrocholecalciferol, 21 [QD +99.60 (c 0.25), MeOH).

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 0 C is 15 added dropwise a solution of 0.180 g of the product of Example Ih) in 13 ml of THF. The mixture is heated at 68 0

C

for 16 hours and recooled at OC. After dilution with 13 ml of ether and addition of 0.30 ml of water and 0.26 ml of 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 to give 0.179 g of [1(R*),1(3E),3a3,4a,- 7 aa)]-(3a,4,5,6,7,7a-hexahydro-l-(5-hydroxy-1.5-dimethyl- 21 -3-hexenyl)-7a-methyl-IH-inden-4-ol, (aD +11.50 (c 0.33, CHC1 3 b) To a solution of 0.120 g of the product of a) in 10 ml of methylene chloride are added 0.500 g of pyridinlum dichromate and 25 mg of pyridinium p-toluenesulfonate. The mixture 14 stirred for 135 minutes. After addition of 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 CuSO 4 water, half saturated aqueous NaHCO 3 and saturated brine. The organic phase is dried and evaporated.

The residue is chromatographed on silica gel with 35% ethyl acetate-hexane to give 90 mg of 7aa)]-3,3a,S.6,7,7a-hexahydro-1-(5 dimethyl-3-hexenyl)-7a-methyl-4-inden-4-one, [a]D2 +30.60 (c 0.17. CHd 3 c) By treating 0.099 g of the product of b) as described in Example lj). there are obtained 0.111 g of -dimethyl-5-Litrimethylsilyl)oxy]-3-hexenyl)-7a-methyl- -41-inden-4-one, 2 +26.40 (c 0.22, CHC 3 d) As described in Example 1k). starting from 0.265 g of (3S-(lZ,3xL,51) 2-[2-.3,5-bisEE(1,1 -dimethyl)dimethyltoo#silyl] oxy] -2-methylenecyclohexylidene] ethyl2Jdiphenyloxide and 0.095 g of the product of there are obtained 0.162 g of (13,3c,5,7E,23E)-1,3--bisE(1,I -9,10-secocholesta-S,7,10(19),16,23-.pentaene, MS m/e 712 e) By treating 0.159 g of the product of d) as in Example C, 11), there are obtained 0.077 g (84) of 1,25-dihydroxy- 24 16,23E-btsd%4ehydrocholecalciferol, D +46.50 (c 0.20, MeOH).

Example 4 a) As described in Example 1k), starting from 0.225 g of (5S-(lZ)i-.(2-[5-([(1.l-dimethylethyl)dlmethylsilylJoxyj -2-methylenecyclohexylidenej ethyljdiphenylphosphine oxide and 0.110 g of the product of Example 3c), there are obtained 0.150 g of (313,5Z,7E,.23E)-3-CC(l,ldimethylethyl)-dlmethylsllyljoxy] -25K (trimethylsilyl).

oxyl-9,10-secocholosta -S,7,10(19),1.Lc,23-pentaene, Ca 4+68.30 (c 0.18, CHCt 3 D 3) -26 b) By treating 0.144 g of the product of a) as described in Example 11). there are obtained 0.076 g of 25-hydroxy-16, 23E-bisdehydrocholecalciterol, 22 1aD+62.50 (c 0.20, Me0OI).

Example a) To a solution of 6.25 g of ethyl 3-bromopropionate in 28 ml of TH-F at -20 0 C are added 28.8 ml of 2.8M methylmagnesium bromide in ether. The mixture is stirred at room temperature for 170 minutes. After addition of 1.5 ml of I saturated aqueous ammonium chloride and of 42 ml of IN E-Cl, the organic phase is separated and the aqueous phase extracted with ether. The organic extr4.ts are washed with saturated brine, dried and evaporated. The residue is chromatographed on silica gel with 30% ethyl acetate-hexane to give 2.57 g of 4-bromo-2.-methyl-2-butanol, MS rn/c 2.51 CH 3 b) To a solution of 2.56 g of 4.-bromo-2-methyl-2-butano].

and 4.86 g of imidazole in 15 ml of N,N-dimethylformamlde at 0 0 C are added 6.48 g~ of chlorotriethylsilane. The mixture to stirred at room temperature for 200 minutes. After adding Ice, the mixture is dilutedI with water and extracted with pentane. The organic phase is washed with water and saturated brine, dre and evaporated. The residue is chromatogrpahed on silica gel with pentane to give 4.02 g of (3-bromo-1,2-dimethylpopoxy)triethylsilane, MS rn/e 265 (M -CI-1 3 c) To a solution of 0.930 g of t2(R*),3aR*(3aJ3,4a, 7ac))-3a,4,5.6,7,7a-hexahydro -4-hydroxy413,7a-dimethyl- 3I-i-ndene-l--ethanol 4-methyl-berezenesulfonate and 1.2.0 g of imidazole in 73 ml of mothylene chloride at 0OC are added 0.580 g of chlorotriethylaitlane. The mixture is stirred at room temperature~ 1 .5 hours. After adding Ice, the muixture is diluted with water and stirred tor 20 minutes.

27 The organic layer is extracted with methylene chloride. The extracts are washed with water, IN N 2 S 0 4 satucated aqueous NaFICO and saturated brine. After drying and 3 evaporation, the residue is purified on silica gel with ethyl acetate-hexane to afford 1.22 g (100%) of C1(R*),3aR*-(3a,4,7a)-3a,4,5,6,7,7a-hexahydro -4-[(triethylsilyl)oxy]-13,7a-dimethyl-3H-indene-1-ethanol 20 4-methylbenze sulfonate, [aD +46.10 (c 0.31,

D

CHCl 3 d) To a solution of 3.08 g of (3-bromo-1,1-dimethylpropoxy)triethylsilane in 31 ml of THF are added 0.282 g of magnesium. The mixture is heated at 68 0 C for 3.5 hours. Then *qrq o a mixture of 0.686 g of cuprous iodide and the above mentioned Grignard solution are stirred at 3 0 C for minutes. To this is added a solution of 1.02 g of the product of 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 otther. The organic phase is washed with 1N H 2 S0 4 and saturated aqueous NaHCO 3 dried and evaporated. The residue is chromatographed on silica gel with ethyl acetate-hexane (1:15) to afford 1.80 g of 7a)] J 3a,4,5,6,7, 7a-hexahydro- -1-[1,5-dimethyl-5-[(triethylsilyl)- oxyjhexylj- -4-[(triethylsilyl)oxy]-7a-methyl-3l-indene, MS m/e 479 (M'-Et) e) To a solution of 1.60 g of the product of d) In 5 ml of THEF are added 2.00 ml of LM tetrabutylammonium fluoride In THF. The mixture is heated at 68 0 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 to afford 0.420 g of [1(R*),3aR*-(3ab,4a, 7a )3-3a,4,5,6,7,7a-hexahydro-4-hydtoxy-a,c.ac,7a- 21 totramethyl-1H-indene-1-pentanol, taL +12.00 -r -r 28 (c 0.25,

CHC

3 f) To a solution of 0.210 g of the product of e) in 18 ml of methylene chloride are added 0.870 g of pyridinium dichromate and 44 mg of pyridinium p-toluenesulFonate. The mixture is stirred for 175 minutes. After addition of 50 ml of ether, the mixture is stirred for 5 minutes and filtered.

The solids are washed with saturated aqueous CuSO 4 water, half saturated aqueous NaFICO 3 and saturated brine. The organic phase is dried and evaporated. The residue is chromatographed on silica gel with 35% ethyl acetate-hexane to give 0.175 g of [1(R*),3aR*-(3aj3,7aa)]- 3,3a,5,6,7,7a-hexahydro--(5-hydroxy-1,5-dimethylhexyl)- 21 7a-methyl-4H-inden-4-one, +28.20 (c 0.22,

D

CHCl

C~

3 g) By treating 0.168 g of the product of f) as described in Example 1j), thece are obtained 0.211 g (100%) of 4 4 4 1 R 3aR*-(3a 3jR a ,3 a, 5, 6, 7, 7a -he xahyd Ioo 1, dimethyl-5-[(trimethylsilyl)oxyhexyl)-7a-methyl-4-inden.

20 4-one, [l20+21.90 (c 0.27, CHC1 3 4,t h) As described in Example 1k), starting from 0.581 g of [3S-(1Z,30.,51-2-3,5-bisC((1,1-dimethyl)dimethylsilyl]oxy]-2-methylenecyclohexylideneJethylidiphenyl phosphine oxide and 0.210 g of the product of there are obtained 0.358 g of (la,31,5Z,7E)-l,3-bisE(1,1-,dmethyl.

ethyl)dimethylsilyljoxyJ- 25- (trimethylsilyl)oxyj-9, secocholesta-57,10(19),16-tetraene, MS m/e 714 i) Treating 0.350 g of the product of h) as described in Example 11), there are obtained 0.168 g of 1,25-dihydroxy-16-dehydrocholecalciferol, 40.00 (o 0.17, MeOH).

-29 Example 6 a) As described in Example 1k) starting-from 0.383 g of [5S-(1Z)J--2-[5-[[(1,1-dimethylethyl)dimethylsilyljoxy)- 2-methylenecyclohexylidene] ethyl ]diphenylphosphine oxide and 0.188 g of the product of Example 5g), there are obtained 0.245 g of (3a,5Z.7E)-3-E(1,1-dimethylethyl)- (trimethylsilyl)oxy]-9,10-secocholesta-5,7,10(19),16-tetraene, 2 +67.50 (c 0.20, CHCl 3 Vb) Treating 0.239 g of the product of a) as described in Example 11) affords 0.135 g of 25-hydroxy-26-dehydro- 23 The following Example A and B illustrate the comtposition of soft gelatine capsules for oral administration and of a topical cream: Example A mg/capsule Compound E 0.0001-0. 010 Butylated hydroxytoluene 0.016 Butytated hydroxyanisole 0.016 Fractionated coconut oil 160.0 30 Example B Comppound E Cetyl alcohol Stearyl alcohol Sorbi tar monos tearate Glyceryl monostearate and polyoxyethylene glycol stearate Polysorbate 60 Mineral oil Propylene glycol Propylparaben Butylated hydroxyanisole Sorbitol solution Edetate disodium Methylparaben Distilled water mg_/g of cream 0.001-1.0 2. 0.05 0.05 0.01 0.18 q.s. to 100 g ti t I tag' r f #1

Claims (9)

1. A compound of the formula -I A A H H HO R wherein R is hydrogen or hydroxy and A is -CsC-, -CH=CH- with E-configuration or -CH 2 -CH 2

2. A compound in accordance with claim 1, of the group consisting of: 1,25-dihydroxy-16-dehydro-23-didehydrocholecalclferol, 1,25-dlhydroxy-16,23E-bisdehydrocholecalciferol and 1,25-dihydroxy-16-dehydrocholecalciferol.

3. A compound In accordance with claim 1 or 2 for use as a therapeutically active substance, particularly for the treatment of hyperprolffratlve diseases of the skin, especially psoriasis, or for the treatment of neoplastic diseases, especially leukemia.

4. A process for the preparation of a compound according to claim 1 or 2, which process comprises reacting a corresponding compound of formula I containing Instead of the two or three hydroxy groups two or three protected hydroxy groups of the formula -OSI(R ,R 2 ,R 3 wherein R I and R 3 are C -4-alkyl and R 2 is C 1 -4-alkyl, aryl or aryl-C 1 -4-alkyl, with an agent capable of removing the protecting groups. 1476 c~- ,4 32 A medicament for the treatment of hyperproliferative diseases of the skin or for the treatment of neoplastic diseases comprising an effective amount of a compound according to claim 1 or claim 2 and a pharmaceutically acceptable carrier, diluent and/or excipient.

6. A method for the manufacture of a medicament for the treatment of hyperproliferative diseases of the skin or for the treatment of neoplastic diseases which comprises admixing a compound according to any one of claims 1 to 3 with a pharmaceutically acceptable carrier, diluent and/or excipient.

7. A method for the treatment or prophylaxis of hyperproliferative skin disease or neoplastic disease in a patient requiring said treatment or prophylaxis, which method comprises administering to said patient an effective amount of at least one compound according to any one of claims 1 to 3, or of a medicament 15 according to claim

8. The method of claim 7 wherein the hyperproliferative skin disease is psoriasis.

9. The method of claim 7 wherein the neoplastic disease is leukemia.

10. 16-Dehydro-vitamin D 3 -derivatlves whenever produced by the process of claim 4. DATED this TWENTY-FIRST day of JANUARY 1992 F Hoffmann-La Roche Co Aktiengesellschaft Patent Attorneys for the Applicant SPRUSON FERGUSON TMS 76u wA