CA2200480C

CA2200480C - New use of prostaglandins

Info

Publication number: CA2200480C
Application number: CA002200480A
Authority: CA
Inventors: Johan Stjernschantz; Bahram Resul
Original assignee: Synphora AB
Current assignee: Synphora AB
Priority date: 1994-09-21
Filing date: 1995-09-19
Publication date: 2003-03-25
Anticipated expiration: 2015-09-19
Also published as: AU3581795A; EP1310256A2; DE69535081T2; EP1310256B1; DE69530303D1; EP1310256A3; EP0778774A2; EP0778774B1; JP4044611B2; DE69535081D1; DE69530303T2; JPH10505861A; ATE236638T1; SE9403158D0; US6031001A; ATE330613T1; CA2200480A1; AU705901B2; ES2194055T3; WO1996009055A1

Abstract

Composition comprising a therapeutically active and physiologically acceptab le amount of prostaglandin A or prostaglandin J or a derivative of any one of these in a carrier, for treatment of psoriasis.

Description

WO 96/09055 ~ ~ ~ o ~ ~ ~ PCT/SE95/01059 New use of prostaglandins.
Psoriasis is a common dermatologic disorder affecting 1-2~ of the population e.g. in Europe and United States.
The disease usually debuts between the age of 10-40 years, but may become manifest at any age. Typically hyperkeratotic pink lesions covered by adherent silver-white scales can be found in patients suffering from psoriasis. The lesions have a characteristic shape and are well-demarcated. Not infrequently these lesions are localised to the elbows, knees, the gluteal regions and the scalp and it is generally believed that a cause of the psoriatic lesions is physical contact, pressure e.g.
rubbing.
The underlying mechanism of psoriasis is an increased proliferation of cells in the epidermis, primarily the keratinocytes. Thus, the epidermis becomes thick and hyperkeratotic, particularly superficially. The precise mechanism behind the stimulus of the cell proliferation is not known, but generally it is believed that trauma of the skin leads to an inflammatory reaction involving hyperproliferation of keratinocytes in the epidermis.
There is a marked genetic disposition to develop psoriasis. Psoriasis may also become generalised over the whole body and psoriasis may cause arthritis, typically in the fingers. Psoriasis may fluctuate but complete and permanent remission is uncommon.
Psoriasis is usually treated with different medications. In simple cases, keratolytics, lubricants and topical corticosteroids are employed. Salicylic acid and ' anthralin are also used. Another form of medical treatment is PUVA-treatment. PUVA-treatment is based on systemic or local administration of psoralens, e.g. methoxy-psoralen combined with irradiation of the skin with ultraviolet light (UVA). This treatment modality is effective but may predispose to skin cancer. Antimitotics, such as WO 96/09055 ~ ~ ~ ~ ~ ~ ~ 2 PCT/SE95/01059 methotrexate, have also been used in severe cases of psoriasis. Although there are presently many treatment modalities for psoriasis there is a definite need for more effective medications with less side-effects.
We have now unexpectedly found that certain prostaglandins may be useful for the treatment of psoriasis. Prostaglandins are fatty acids usually derived from the precursors eicosatrienoic, eicosatetraenoic or eicosapentanoic acid through metabolic steps involving oxygenation. Naturally occurring prostaglandins typically have the general structure ,,. _ X COON

11 ~3 v v "20 OH
The prostaglandins accordingly carry a cyclopentane ring to which two carbon chains link, the upper usually being called the alpha chain and the lower usually being called the omega chain.
The prostaglandins are classified in subgroups A, B, C, D, E, F and J depending on the structure of the cyclopentane ring:
O O O OH
A B C O D
O OH
~~~i ,iii \ ' \
OH E OH F O

~~ao~~ o The alpha chain is a 7 carbon carboxy-terminated aliphatic chain whereas the omega chain is a 8 carbon methyl-terminated aliphatic chain. Depending on the number of double bonds in these chains subscripts of 1 to 3 are given. In prostaglandins with subscript 1, e.g. PGA1 and PGJ1, the double bond is situated between carbons 13 and 14 in tie omega chain, and it exhibits trans configuration in naturally occurring prostaglandins. In prostaglandins with subscript 2, e.g. PGA2 and PGJ2 an additional double bond in the cis configuration exists between carbons 5 and 6 in the alpha chain and finally in prostaglandins with subscript.3 a third double bond is situated between carbons 17 and 18 in the omega chain. This double bond also exhibits cis configuration in naturally occurring prostaglandins. All naturally occurring prostaglandins carry a hydroxyl group in carbon 15, which is essential for biologic activity.
Therapeutic use of prostaglandins for treatment of a great number of various diseases, including psoriasis, has been suggested, especially in patent publications, but no efficient prostaglandin derivative has to be best of our knowledge been presented for treatment of psoriasis.
Prostaglandins to be used according to the present invention are characterized by an a,~i-unsaturated cyclopentenone and are in particular of the A and J type in which the cyclopentene ring has the basic structure.
O
,. ,, I
O
PGA PGJ
The prostaglandins that have been utilized in the exemplification of the present invention are PGA2 and PGJ2. PGA2 is probably not a naturally occurring prostaglandin in man, but it is formed from PGE2 during acid extraction. PGJ2 on the other hand is a well known metabolite of PGD2, which is a naturally occurring prostaglandin. The molecular structures of PGAzand PGJz are depicted in the figure given below.
O , ~~~~~ COO H
OH

~_ J/~ C00H
_ _ O OH

Exemplification of the invention The invention is exemplified with the following non-limiting examples. Prostaglandin AZ ((5Z, 13E, 15S)-15-hydroxy-9-oxoprosta-5, 10, 13-trim-1-oic acid) and prostaglandin JZ (5Z, 13E, 15S)-15-hydroxy-11-oxoprosta-5, 9, 13-trien-1-oic acid) were purchased from Cayman Chemical Company (Ann Arbor, Michigan, USA) and used in acid form. Both compounds were dissolved in ethanol, and diluted to the final concentration directly in the cell growth medium.
Normal human epidermal keratinocytes (NHEK) derived from foreskin were purchased as secondary cultures produced by PromoCell GmbH of Heidelberg, Germany, hereinafter simply "PromoCell" and cultured in an optimized ready-to-use serum-free growth medium (KGM
medium) (PromoCell) at 37°C in 5~ CO2, humidified air.
The growth medium is a modification of the MCDB 153 formulation and is supplemented with various concentrations of human epidermal growth factor, insulin, hydrocortisone, bovine pituitary extract and gentamicin/amphotericin B (proportions proprietary information of PromoCell) .

For the experiments, cells at passage 3 were used.
The effects of the prostaglandins were examined with a photometric cell proliferation assay after 5 days of continuous exposure. Cells were seeded into multiwell tissue culture plates containing KGM medium and quadruplicates of 50 nanomolar to 50 micromolar of the test compounds. KGM medium only, served as control. Every second day the culture medium was exchanged with fresh medium including the appropriate concentration of fresh prostaglandin to provide the cells with sufficient nutrients and to avoid problems that could arise from degradation of the test compound in the culture medium.
After 5 days the cells were fixed in glutaraldehyde (1~) and stained by crystal violet (0.1~) whereupon the stain was eluted by sodium lauryl sulphate (2.5~). The absorbance of the colored solutions, shown to the be linearly related to cello number, was monitored photometrically. The experiment was repeated once in its entirety.
The test results showed that both PGAZ (compound x) and PGJ2 (compound y) markedly inhibited cell growth and reduced the total number of cells. The growth inhibitory effects of the test compounds were noted microscopically after 48 hours, the first time point of observation. For PGAZ (compound x), a decreased cell density but normal cell morphology, as compared to the control, was found in the wells containing 5 micromolar. At 25 and 50 micromolar most cells were still attached to the substratum but all cells were pyknotic, i.e. very small and irregular. For PGJZ (compound y), a reduction in the number of attached cells but normal cell morphology was found in the wells containing 0.5 micromolar. At 5 micromolar and higher concentrations all cells appeared pyknotic.

After 5 days the growth inhibitory effects of both compounds were even more pronounced. Both PGAz (compound x) and PGJz (compound y) markedly suppressed growth and reduced the total number of cells in a dose-dependent manner. The highest concentrations tested reduced the cell numbers by 90 ~. PGJz was the most potent compound and exerted a half-maximal growth inhibition at about 0.2 micromolar concentration. The corresponding value of PGAZ was about 1 ,uM. The control (compound z) which was the vehicle of the prostaglandin solutions had no effect on cell growth. Thus, both PGAz and PGJz had marked inhibitory effect on the cultured human keratinocytes.
It is accordingly clearly indicated that prostaglandins of the A and J type may be utilized for the treatment of psoriasis. In the exemplification only two prostaglandins, namely PGAz and PGJz were used, but analogues and derivatives of prostaglandins of the A and J type with the same fundamental mechanism of action may also be employed. Analogues of PGA includes e.g. 16,16-dimethyl-PGA1, o' -PGJ1, o' -PGAZ and 16, 16-dimethyl-PGAZ.
Analogues or derivatives of PGJ include e.g. PGAI, olzpGJ1 and olzPGJ2.
There are also other types of derivatives of the A
and J type which are known from the literature and which are obvious candidates to be used for treatment according to the present invention. One such group is the derivatives containing a ring substituted omega chain disclosed in PCT publication WO 90/02553. Also alpha-chain modified prostaglandin may be employed, for instance derivatives containing alkyl substituents.
PGA and PGJ or their analogues may be modified to more lipophilic substances by esterification of different WO 96/09055 ~ ~ ~ ~ ~ ~ ~ ~ PCTISE95/01059 parts of the molecule, e.g. the carboxylic acid moiety.
Such esters that may be employed clinically because they penetrate better into the skin comprise alkyl esters with 1-10 carbon atoms and especially short alkyl esters e.g.
methyl, ethyl, and isopropyl or cyclic esters such as benzyl.
The prostaglandin compounds and their esters or derivatives should be used in a suitable vehicle for topical application on the skin. A suitable vehicle includes aqueous vehicles with or without solubilizers, stabilizers such as cyclodextrins, oils, ointments, micelle systems, nanoparticles and various slow release formulations. Such vehicles may or may not contain preservatives depending on whether they are intended for single or multiple use. Various preservatives that may be employed comprise e.g. benzalkonium chloride, chlorhexidine, thiomersal, parabenzoic acid and other compounds with satisfactory antimicrobial effect.
Accordingly, in one aspect of the present invention a formulation containing PGA2 or PGJ2 or derivatives of these prostaglandins is applied topically on the affected skin for different periods of time once or several times daily to treat the psoriatic lesions. Such treatment may take only a few weeks or may go on for longer times depending on the clinical situation. The recommended dose to be used depends on the particular prostaglandin and its physical-chemical characteristics but is usually in the range of from 0.01 to 100 ug per application. On an area of 1 dm2 typically a dose of 0.1-10 ug per application is employed. The medication can be instilled once or several times daily depending on the clinical situation, and the dosage form. L~hen the psoriatic lesion has regressed ' treatment may continue intermittently or may be terminated.

WO 96/09055 ~ ~ ~ ~ 8 PCT/SE95/01059 The invention also relates to the use of a a prostaglandin derivative as defined above for the preparation of a composition for treatment of psoriasis.

Claims

CLAIMS:

1. Use of a prostaglandin or prostaglandin derivative selected from the group consisting of therapeutically active and physiologically acceptable prostaglandin A2, prostaglandin J, derivative of prostaglandin A2, derivative of prostaglandin J, an alkyl ester of prostaglandin A having 1-10 carbon atoms in the alkyl group, and benzyl ester of prostaglandin A, for the topical treatment of psoriasis.

2. Use of a prostaglandin or prostaglandin derivative selected from the group consisting of therapeutically active and physiologically acceptable prostaglandin A2, prostaglandin J, derivative of prostaglandin A2, derivative of prostaglandin J, an alkyl ester of prostaglandin A having 1-10 carbon atoms in the alkyl group, and benzyl ester of prostaglandin A, for inhibiting an increased poliferation of cell growth associated with psoriasis in epidermis.

3. Use of a prostaglandin or prostaglandin derivative selected from the group consisting of therapeutically active and physiologically acceptable prostaglandin A2, prostaglandin J, derivative of prostaglandin A2, derivative of prostaglandin J, an alkyl ester of prostaglandin A having 1-10 carbon atoms in the alkyl group, and benzyl ester of prostaglandin A, for the manufacture of a medicament for the treatment of psoriasis.

4. Use according to claim 1, claim 2 or claim 3, wherein said prostaglandin or prostaglandin derivative is a prostoglandin A2 or a derivative thereof.

5. Use according to claim 1, claim 2 or claim 3, wherein said prostaglandin or prostaglandin derivative is of an alkyl ester of a PGA having 1-10 carbons in the alkyl group or a benzyl ester of a PGA, or a derivative thereof.

6. Use according to claim 1, claim 2 or claim 3, wherein said prostagladin or prostaglandin derivative is a methyl, ethyl, or isopropyl ester of a PGA, or a derivative thereof.

7. Use according to claim 1, claim 2 or claim 3, wherein said prostagladin or prostaglandin derivative is an isopropyl ester of a PGA, or a derivative thereof.

8. Use according to claim 1, claim 2 or claim 3, wherein said prostagladin or prostaglandin derivative is a protaglandin J or a derivative thereof.

9. Use according to claim 1, claim 2 or claim 3, wherein said prostaglandin or prostaglandin derivative is PGJ1, PGJ2, .DELTA.12PGJ1, .DELTA.12PGJ2, a ring-substituted omega chain PGJ, an alkyl-substituted alpha chain PGJ, or a PGJ
ester, or a derivative thereof.

10. Use according to claim 1, claim 2 or claim 3, wherein said prostaglandin or prostaglandin derivative is an alkyl ester of a PGJ having 1-10 carbons in the alkyl group or a benzyl ester of a PGJ, or a derivative thereof.

11. Use according to claim 1, claim 2 or claim 3, wherein said prostaglandin or prostaglandin derivative is 11~

a methyl, ethyl or isopropyl ester of a PGJ, or a derivative thereof.

12. Use according to claim 1, claim 2 or claim 3, wherein said prostaglandin or prostagaldin derivative is an isopropyl ester of a PGJ, or a derivative thereof.