IE64186B1

IE64186B1 - Dehydrocholecalciferol derivatives

Info

Publication number: IE64186B1
Application number: IE178090A
Authority: IE
Inventors: Enrico Giuseppe Baggiolini; Bernard Michael Hennessy; Shian-Jan Shiuey; Gary Arthur Truitt; Milan Radoje Uskokovic
Original assignee: Hoffmann La Roche
Priority date: 1989-05-18
Filing date: 1990-05-17
Publication date: 1995-07-12
Also published as: JPH0317019A; EP0398217A1; EP0398217B1; ZA903758B; DE69005897D1; JPH0686381B2; CA2016985C; IE901780L; NZ233704A; ATE99942T1; CA2016985A1; AU636609B2; DK0398217T3; AU5516590A

Abstract

The use of dehydrocholecalciferal derivs. of formula (I) to make pharmaceuticals for treating diseases of the sebaceous glands is new. (Where R = H or OH; A = CC, (E)-CH=CH or CH2CH2; R'' = H or, where A = C=C, it can also be deuterium). (I) are new cpds. where R'' = deuterium.

Description

The invention is concerned with the use of dehydrocholecalciferol derivatives of the formula wherein R is H or OH, A is the grouping -C=C-, -CH=CH- with the E-configuration or -CH2CH2- and R is H or, where A is -C=C-, R is also deuterium, for the manufacture of pharmaceutical preparations for the treatment of sebaceous gland diseases which are associated with a proliferation of sebocytes, such as acne and seborrheic dermatitis.

Among the compounds of formula I, those in which R is deuterium are novel and as such are an object of the invention. The deuterated compounds can be prepared in analogy to the nondeuterated compounds, e.g. as described in European Patent Application No. 325 279 via intermediates of the formula wherein R has the above significance, R1 and R3 are loweralkyl and R2 is lower-alkyl, aryl or aryl-lower-alkyl.

The following compounds A to H are examples of compounds of formula I: A: 1 a,25-dihydroxy-16-dehydrocholecalciferol; B: 25-hydroxy-16-dehydrocholecalciferol; C; 1 a,25-dihydroxy-16,23E-bisdehydrocholecalciferol; D; 25-hydroxy-16,23 E-bisdehydrocho I ecalciferol; E 1 a,25-dihydroxy-16-dehydro-23-didehydrocholecalciferol; F: 25-hydroxy-16-dehydro-23-didehydrocholecalciferol; G 26,26,26,27,27,27-hexadeutero-1 a ,25-di hydroxy -1 6dehydro-23-didehydrocholecalciferol and H; 26,26,26,27,27,27 -hexadeutero-25-hydroxy-16-dehydro23-didehydrocholecalciferol.

The efficacy of compound of formula I in the treatment of acne can be demonstrated as follows; Sebocytes were isolated from adult human sebaceous glands in analogy to the method described in Cells 6, 1975, 331-334 and In Vitro 22:3, 1986, II, p. 22a, abstract 46 and cultured on a layer of mouse fibroblasts 3T3. A suspension of sebocytes was prepared from skin tissue by enzymatic and mechanical methods. Thus, the cells were cultured either in Isocove's medium containing 2% human serum, 8% foetal calf serum and 4 μg/ml of dexamethasone or in Isocove's medium containing 10% foetal calf serum and 4 μρ/ΓηΙ of dexamethasone.

The test compounds in the medium were added to the cultures every 48 hours. After the last addition the fibroblasts 3T3 were rinsed with 0.03% EDTA in PBS. The sebocyte colonies remaining behind were incubated in 0.05% trypsin/0.03% EDTA with the formation of a uniform cell suspension of sebocytes.

The cells were then diluted and counted in a haemocytometer. ’2 ethanolic solutions of the test compounds were prepared and then diluted to 10’6, 10*7, 10'8and 10'θΜ. 1aDihydroxycholecalciferol (compound X in the following Tables) was also tested together with the compounds of formula I for the inhibition of the proliferation of sebocytes in vitro.

The results are given in Table 1 hereinafter as the amount of compound which inhibits the proliferation of the sebocytes by 50% compared with a control value. The control value was obtained from a cell culture treated with diluent only.

Table 1 Compound_X_APE_F ED50 (μΜ) 0.005 0.001 >1 0.001 0.1 The results demonstrate that the compounds of formula I inhibit the proliferation of human sebocytes in vitro and therefore are useful as agents for the treatment of acne.

The topical efficacy of compounds of formula I against acne was evaluated on hamster ear sebaceous glands. 50 μΙ of a solution of the test compound were applied daily to the ear of test animals and 50 μΙ of acetone were applied to control animals. The animals were killed after 4 weeks. The areas of the sebaceous glands on the ears were determined. The results are given in Table 2 as the % change compared with control animals of the cross section of hamster ear sebaceous glands.

Table 2 Compound Dose (pg/hamster) % Change D 0.10 -1 6 1.00 -16 10.00 -43 F 0.01 - 8 0.10 -23 1.00 -40 .00 -64 In order to evaluate the calcification of soft tissues by the compounds of formula I, rats received subcutaneous injections of 40 pCi 45Ca. The compounds were then administered sub- cutaneously or topically for 4 days. The rats were killed 1 day after the last injection. The hearts and kidneys were removed and treated with nitric acid. The radioactivity of an aliquot of 0.2 ml of this preparation was measured. The calcification ratio (com of X-control com) (cpm of test compound-control cpm) is given in Table 3: Table 3 Compound Subcutaneous Topical X 1 1 D >1400 >34 E 47 >1 F >1400 >34 The effect of compounds of formula I on hamster ear sebaceous glands after oral administration was evaluated. 200 μΙ of a solution of the test compounds in propylene glycol were administered daily to male Syrian golden hamsters. The animals were killed after 4 weeks. The area of the sebaceous glands on the ears was measured. The results are given in Table 4 as the change in cross section of the hamster ear sebaceous glands.

Table 4 Compound Dose (pg/hamster) % Change D 2.50 -1 5‘ 10 5.00 -22" 10.00 -27* " 20.00 -36*" F 0.05 -1 6* 0.50 -23** 15 5.00 -42* " 50.00 -55*** * p <0.05; **p <0.01; *** p <0.001 The above results demonstrate that compounds of formula I are useful as active substances in the treatment of sebaceous gland diseases, such as acne or seborrheic dermatitis, and, moreover, that they bring about less soft tissue calcification than 1 a,25-dihydroxycholecalciferol. Soft tissue calcification is an undesirable side effect of an agent for the treatment of sebaceous gland diseases.

The compounds of formula I can be administered topically or orally for the treatment of sebaceous gland diseases, such as acne or seborrheic dermatitits. Thus, they can be administered in an oral dosage of about 0.7 to 700, preferably 7.0 to 70, pg per day for the oral treatment of acne.

For oral administration, the compounds of formula I may be incorporated e.g. into capsules or tablets together with pharmaceutically usable carriers. Examples of such carrier materials for capsules are binders, such as tragacanth, gums or gelatine; excipients, such as dicalcium phosphate; disintegrating agents, such as corn starch; lubricants, such as magnesium stearate; sweetening agents, such as sucrose; flavouring 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 flavouring agent.

Topical dosage forms containing compounds of formula I are ointments and creams consisting of oleaginous, absorbable, water-soluble and emulsion-type bases, such as lanolin and polyethylene glycols. Other topical dosage forms are gels, lotions, powders and aerosols.

Lotions, i.e. liquid preparations, such as solutions or aqueous or hydroalcoholic preparations, which contain the substance in powder form, can contain, in addition to the active substance, suspending or dispersing agents, such as cellulose derivatives, e.g. ethyl or methyl cellulose; gelatine or gums, as well as a vehicle, such as water, alcohol or glycerol. Gels are semi-solid preparation which are produced from a solution or suspension of the active substance in a vehicle. The aqueous or anhydrous vehicles are treated with a gelling agent, e.g. carboxypolymethylene, and neutralized with a base, e.g. sodium hydroxide or an amine such as polyethylenecocoamine.

The composition of soft gelatine capsules for oral administration and of a cream for topical administration is given in the following Examples: Example A mg/caps-ui.e Compound E Butylated hydroxytoluene Butylated hydroxyanisole Fractionated coconut oil 0.0001-0.010 0.016 0.016 160.0 Example Β Compound Ε mp/ cream 0.001-1.0 Λ Cetyl alcohol 1.5 5 Stearyl alcohol 2.5 Sorbitan monostearate Glyceryl monostearate and 2.0 polyoxyethylene glycol stearate 4.0 Polysorbate 60 1.0 10 Mineral oil 4.0 Propylene glycol 5.0 Propylparaben 0.05 Butylated hydroxyanisole 0.05 Sorbitol solution 2.0 15 EDTA disodium salt 0.01 Methylparaben 0.18 Distilled water q.s. to 100 g

Claims

1. The use of dehydrocholecalciferol derivatives of the formula wherein R is H or OH, A is the grouping -C=C-. -CH=CH- with the E-configuration or -CH2CH2- and R is H or. where A is -C=Q·. R is also deuterium, for the manufacture of pharmaceutical preparations for the treatment of sebaceous gland diseases which are associated with a proliferation of sebocytes.

2. The use according to claim 1, wherein the disease to be treated is acne.

3. Compounds of formula I in claim 1. wherein R is deuterium.

4. Use according to claim 1, substantially as hereinbefore described and exemplified.

5. A compound as claimed in claim 3, substantially as hereinbefore described and exemplified.