MXPA99004182A - Synergistic gold-containing compositions - Google Patents

Abstract

This invention relates to a method of treating an immune-mediated disorder having one or more manifestations. The method comprises administering to a patient requiring such treatment a gold compound and at least one corticosteroid, wherein the at least one corticosteroid is selected to interact synergistically with the gold compound to exhibit preferential action towards one of the manifestations of said disorder or to exhibit equal action towards each manifestation of said disorder. The invention also relates to a pharmaceutical composition suitable for use in the method.

Description

SYNERGISTIC COMPOSITIONS CONTAINING GOLD TECHNICAL FIELD The present invention relates to pharmaceutical compositions that include a gold compound in combination with a corticosteroid and its use in the treatment of dermatological disorders.

BACKGROUND OF THE INVENTION The effectiveness of gold compounds in the treatment of rheumatoid arthritis has been known since the 1960s. More recently, gold complexes have been used as therapeutic agents in the treatment of rheumatoid arthritis but their exact mechanism remains unknown. The most commonly used complexes have been gold thiolates (Au (l)) soluble in water administered parenterally, such as aurothiomalate (Myocrisin®) and aurothioglucose (Solganol®). Subsequently, a number of alkylphosphine and gold complexes showed anti-arthritic activity when orally administered to auxiliary arthritic rats. The auranofin compound (1-thio-β-D-glucopyranose 2,3,4,6-tetraacetate-S) - (triethylphosphine) gold Au (l)) was among the most potent and effective compounds that were tested and are currently being used for the treatment of rheumatoid arthritis in men.

The gold compounds have been administered by intravenous and oral routes for the treatment of asthma, tuberculosis, pemphigus vulgaris, various forms of arthritis, cancer and infection. However, treatment with gold compounds has been frequently associated with unacceptable and sometimes serious side effects. Corticosteroids have found similar therapeutic applications. The success of topical corticosteroids in the therapy of inflammatory and proliferative skin disorders has led to the vigorous development of new corticosteroids since their first topical use. An increase in potency has been achieved by chemical modification of the natural corticosteroid, hydrocortisone, without knowing precisely the mechanism of action of corticosteroids. The development of more potent corticosteroids has extended its usefulness in a wide variety of skin disorders, but especially with long-term use, has led to undesirable effects. Systemic effects such as hypothalamic pituitary-adrenal axis depression were previously known to arise from the systemic use of corticosteroids. Local side effects after topical application were observed only with the most potent synthetic spheroids. The most common serious side effects of topical corticosteroids are thinning of the skin, stretch marks and telangiectasia. During long-term treatment with very potent corticosteroids, the inflammatory cells are affected and in addition the proliferation of keratinocytes and the activity of the fibroblasts are inhibited.

Fibroblasts synthesize important structural and functional components of the skin, namely collagen, elastin and glucosamino-glycans. The inhibition of keratinocyte proliferation leads to a thinning of the epidermis. Although the effect on the epidermis is normally reversible, the dermis can be damaged irreversibly. Recently, topical formulations of organic gold colas have found utility for the treatment of skin disorders such as psoriasis. In this way, the Australian patent no. 616,755, describes the use of a topical formulation of auranofin in combination with a corticosteroid for the treatment of local inflammatory conditions such as those associated with psoriasis. In particular, treatment with a formulation including auranofin and betamethasone dipropionate demonstrated a remarkable synergistic action when compared with the same concentrations of the individual active ingredients. The finding that a gold compound can synergize to a corticosteroid allowed the use of considerably lower levels of both the gold compound and the corticosteroid in the formulations, thereby allowing more effective therapy while avoiding the well-known side effects associated with the use of either gold compounds or corticosteroids alone. Dermatological disorders are frequently associated with different manifestations of inflammation only. For example, psoriasis also contains a component of cellular hyperproliferation (hyperplasia), whose mechanism is fundamentally different from that of inflammation and thus may not necessarily be affected by topical gold / corticosteroid formulations. In addition, the anti-inflammatory component of different dermatological conditions can vary from very mild to very severe, creating the need for variations in the formulation, in particular the choice of corticosteroid that could allow not only effective and appropriate treatment of the anti-inflammatory component, but also provide differential action in dermatological conditions where there is an additional component such as cell hyperproliferation. Other immune, autoimmune and infectious disorders may also be associated with multiple manifestations, where effective treatment may be based on targeting only one or more of the manifestations of the disorder, depending on the disorder treated and the patient's evaluation. The present invention is based on the surprising finding that there are important differences between corticosteroids with respect to the degree of potentiation of the effects and the type of potentiated effect, when combined with a gold compound. That is, when different corticosteroids are combined with a gold compound, not only is the similarity of synergistic action expected against inflammation achieved, but also a differential synergistic action with respect to inflammation and hyperplasia is demonstrated. In the compositions of the present invention certain corticosteroids synergize with the gold compounds to provide a greater effect on the inflammatory component of a disorder, such as psoriasis, while other corticosteroids give rise to compositions with preferential effects on cellular hyperproliferation. It is contemplated that the compositions of the present invention could also be effectively used for the treatment of a variety of systemic, ti-specific immune, autoimmune and inflammatory disorders.

BRIEF DESCRIPTION OF THE INVENTION According to a first aspect the invention consists of a method for treating an immune-mediated disorder having an inflammatory component and / or a cellular hyperproliferation component, which includes the step of administering to a patient in need of such treatment a gold compound and at least one corticosteroid, wherein the corticosteroid (at least one) is selected to interact with the gold compound to exhibit a preferential synergistic action toward one of the components of said disorder or to exhibit an equal action toward each component of said disorder . According to a preferred embodiment, the present invention consists in a method for treating an immune-mediated disorder according to the first aspect that includes the step of administering at least two corticosteroids, of which at least one is selected to interact with the compound of gold so that a preferential synergistic action is exhibited towards the inflammatory component, and the other component is selected to interact with the gold so that it exhibits preferential synergistic action towards the cellular hyperprophylactic components of said disorder. According to another preferred embodiment, the present invention consists of a method for treating an immune-mediated disorder having multiple components, including the step of administering to a patient in need of such treatment one or more compositions that include a gold compound and one or more corticosteroids, wherein the corticosteroid is selected to deliver a composition with synergistic action equal to each component of said disorder. According to another preferred embodiment, the present invention is a method for treating an immune mediated disorder having one or more components, including the step of administering to a patient in need of such treatment a composition that includes a gold compound and one or more compositions that include one or more corticosteroids wherein the corticosteroid is screened to deliver a composition with preferential synergistic activity toward one of the components of said disorder and wherein the composition that includes a gold compound is orally administered and the composition or compositions that include one or more corticosteroids are administered topically, in effective amounts to supply a gold composition and a corticosteroid having preferential synergistic action towards said component. Preferably the immune-mediated disorder component is an inflammatory component and / or a cellular hyperproliferation component and the composition includes at least two corticosteroids, of which one is selected to deliver a composition with preferential synergistic action towards the inflammatory component and the second corticosteroid is selected to provide a composition with preferential synergistic action toward the cellular hyperproliferation component of said disorder. Preferably the disorder to be treated is an immune-mediated dermatological disorder that is associated with more than one component. Examples of immune-mediated dermatological disorders include psoriasis or dermatitis, such as contact dermatitis, atopic dermatitis or seborrheic dermatitis. Other disorders include rheumatoid arthritis. Typical components of such disorders include an inflammatory component and / or a cellular hyperproliferation component. The corticosteroid can be selected to deliver a composition with synergistic activity toward cellular hyperproliferation in preference to inflammation or vice versa. A corticosteroid as such may be, for example, betamethasone dipropionate, fluocinolone acetonide or hydrocortisone. In cases in which inflammation does not need to be the target in preference to cellular hyperproliferation, the corticosteroid may be betamethasone dipropionate, fluocinolone acetonide or mometasone furoate. The composition is suitably formulated for topical administration. Where the mediated immune disorder is characterized by a number of different components, the preferred method of treatment could use one or more compositions that include a gold compound and one or more corticosteroids, wherein the corticosteroids are selected to deliver compositions with preferential activity to only one of the components of said disorder. In this way the treatment of each individual component is achieved through the use of compositions that include one or more selected corticosteroids, which can be applied in the form of two or more separate compositions or a single composition that includes two or more corticosteroids . Preferably, the gold compounds used in the present invention are liposoluble. Even more preferably, the gold compounds used are formulated for topical application. However, it will be understood that compositions administered systemically or locally are also within the scope of the present invention and include those administered by injection, preferably intra-articularly. In this regard, corticosteroids can be formulated for oral, topical, systemic or local administration. According to a second aspect, the present invention consists of a pharmaceutical composition that includes a gold compound and one or more corticosteroids, the corticosteroid being selected to interact with the gold compound to exhibit a preferential synergistic action towards an inflammatory component and / or a cellular hyperproliferation component of an immune-mediated disorder, in combination with a pharmaceutically acceptable carrier, excipient, auxiliary or solvent.

Preferably the gold compound is auranofin and the corticosteroid is selected from the group consisting of hydrocortisone acetate, hydrocortisone, betamethasone, betamethasone dipropionate, dexamethasone, fluocortolone pivalate 21, triamcinolone acetonide, betamethasone valerate, alclomethasone dipropionate, halcinonide, mometasone furoate or fluocinolone acetonide. More preferably the corticosteroid is selected from the group comprising hydrocortisone, betamethasone dipropionate, mometasone furoate or fluocinolone acetonide.

BRIEF DESCRIPTION OF THE FIGURES Figure 1: A histogram showing the effects of auranofin and glucocorticoids alone or in combination on epidermal hyperplasia induced with TPA. BMD: betamethasone dipropionate; HYD: hydrocortisone; FA: fluocinoione acetonide; MMF: mometasone furoate; AF: auranofin. The bars indicate the standard error of the mean (SEM). Figure 2: A histogram showing the effects of auranofin and glucocorticoids alone or in combination on inflammatory cell infiltration induced with TPA. . BMD: betamethasone dipropionate; HYD: hydrocortisone; FA: fluocinolone acetonide; MMF: mometasone furoate; AF: auranofin. The bars indicate the standard error of the mean (SEM).

DESCRIPTION OF THE PREFERRED MODALITY For convenience a psoriasis model with TPA (O-tetradecanoylphorbol-12 acetate-13) will be used, as an example of an immuno-mediated disorder having an inflammatory component as well as a cellular hyperproliferation component to demonstrate the differential action of different corticosteroids as well as the differential action of different formulations of a gold compound and a corticosteroid. Although psoriasis does not occur in animals other than humans, studies have shown that the application of TPA produces an inflammatory reaction with epidermal thickening that resembles psoriasis in many ways. This produces epidermal hyperplasia and the infiltration of inflammatory cells in the dermis, both of which are typical of psoriasis. TPA increases the activity of the phosphoiipase C / inositol / diacylglycerol triphosphate system. This system activates protein kinase C and the arachidonic acid pathway. Both systems have been implicated in the pathogenesis of psoriasis. It is believed that a mouse treated with TPA is an appropriate model for psoriasis. This animal model will be used to demonstrate the differential actions of different corticosteroids and the synergistic effects of compositions that include a gold compound and a corticosteroid. Three parameters have been measured: thickening of the skin folds, epidermal hyperplasia and infiltration of inflammatory cells in the dermis.

These characteristics, particularly the last two, are the hallmark of psoriasis. However, it is worth noting that the current findings clearly have implications beyond the mere treatment of dermatological disorders. The systemic or inflammation conditions of tissues and hyperplastic could also benefit from the treatment with the compositions of the present invention.

Topical corticosteroids Topical corticosteroids can be grouped according to their strength into: weak, medium, strong and very strong. The vasoconstriction tests are considered as the best method to evaluate the potency of different preparations. It is not known whether the measurement of vasoconstriction predicts anti-inflammatory activity or not. Other evaluation methods available include clinical trials, dermal thickening radiography, biopsy to evaluate epidermal thinning, and mitotic inhibition tests. A useful clinical guide of the relative potencies of topical corticosteroid preparations is shown in Table 1, with the arrangement of the order of ranking approximately the same for ointments and creams. The preparations in each group are only poorly equipotent.

TABLE 1 A guide to the clinical potencies of topical corticosteroids The invention will be described more particularly with reference to the specific embodiments only by way of non-limiting example.

EXAMPLE 1 Treatment of animals and measurement methods Materials Auranofin was kindly donated by Smith Kline and Beecham Pharmaceuticals, King of Prussia, Philadelphia, USA. Alclometasone dipropionate, Betametasone dipropionate, Betametasone valerate, Betametasone (as free alcohol) and Mometasone furoate were kindly donated by Schering-Plow Pty. Ltd., Baulkham Hills, NSW, Australia. Halononide, Hydrocortisone and Triamcinolone acetonide were kindly donated by Bristol-Myers Squibb Pharmaceuticals Pty. Ltd.

The Dexametasona was kindly donated by Roussel Uclaf., Paris, France. Flucortolone pivalate 21 was kindly donated by Schering AG, Berlin, Germany. AND! O-tetradecanoylphorbol 12 acetate, Fluocinolone acetonide, potassium aluminum sulfate, sodium bicarbonate, sodium iodate, magnesium sulfate, eosin Y, floxin, calcium carbonate, acetic acid and Formaidehyde (17 M), thymol, xylene, hematoxylin and "Paraplast" medium for fixing the tissue were obtained from Sigma Chemical Company, Castle Hill, NSW, Australia. Sorbolene cream A.P.F. it was obtained from Wille Laboratories, Carole Park, Queensland, Australia.

Preparation of 2% solution of auranofin Auranofin (20 mg) was dissolved in 10 mL of acetone to obtain the concentration of 0.2%. The auranofin solution was prepared daily for each experiment.

Preparation of auranofin ointment Auranofin ointment was prepared in different concentrations according to the formula as shown below: Concentration auranofina propilenglicol white paraffin (% P / P) (10%) soft 0.20 40 mg 2 g to 20 g 0.50 100 mg 2 g to 20 g Methods of treatment of animals Female BALB / c mice aged 6 to 8 weeks were obtained at the University of Sydney, and treated according to a protocol approved by the Ethics and Animal Care Committee of the University of Sydney. The mice were housed in stainless steel cages, 6 mice per cage, under normal laboratory conditions (room temperature close to 22 ° C) at least 7 days before the experiments for them to acclimate. Food and water ad libutum was allowed throughout the period of the experiment. The backs of the mice were shaved with an electric shearer 2 days before each treatment and only those mice that did not show hair growth were used (that is, the mice that were in the resting phase of the growth cycle were chosen). hair). During the treatment, the mice were held by the tail and placed on the cage so that they could grab the cage and rest there. The solutions were applied in an area of approximately 2 cm x 2 cm on the shaved back of the mice using a "Pipetman" to apply the solution. When an ointment or ointment base ointment was used, the amount was standardized using a micro-spatula that was folded at one end, then the ointment was applied on the spine and the excess was removed by another micro-spatula and applied sparingly twice daily. After a certain time, the mice were sacrificed routinely by cervical dislocation between 9 a.m. and 11 a.m. to avoid variations caused by circadian rhythms, and an area of 1 cm x 1 cm was excised from the center of the treated area by scalpel and scissors. The rest of the tissues were discarded by combustion. The tissues were then fixed, embedded, sectioned and stained.

Methods of preparation of skin sections The preparation method of skin sections was adapted from the method developed by the Department of Pathology, University of Sydney and proved to be successful.

Fixation, embedding, sectioning and dyeing Standard preparation procedures were used. In short, the tissue was fixed in formalin regulated at 10% for 24 hours, washed with tap water for 10 minutes and then processed in an automatic tissue processor (Tissue-Tek VIP 200). The tissue was dehydrated in the automatic tissue processor with graduated series of alcohol and xylene at room temperature, and then infiltrated with 4 paraffin changes ("Paraplast" tissue assembly medium) at 60 ° C. The tissue was finally embedded in fresh paraffin. 5 μm sections were cut with an American Optical Spencer "820" microtome. The sections were then mounted on clean microscope slides using wood glue ("Selleys" Aqueadhere, 1: 100 dilution with water) as an adhesive, and allowed to dry in an oven at 45 ° C for at least 2 hours (usually all night). After the sections were stained blue in Scott's blue dye solution, they were examined under a microscope to confirm that the nuclei were clearly stained and that the cytoplasm did not show staining.

Measurement of skin fold thickness The skin of the back of the shaved mice was folded and measured using a micrometer screw gauge "Etalon". A measurement was taken for each mouse.

Measurement of the thickness of the epidermis The thickness of the epidermis was determined by image analysis.

This image analysis system consisted of a minicomputer (Tracor Northern TN8500) attached to a light microscope (Zeiss Axioplan) and a video camera (Sony DXC-3000P). Sections were taken from each block of tissue and 20 measurements were taken at fixed intervals for each section. The average value of the 20 measurements was obtained and entered as a single value for each mouse. The mean and SEM were calculated for the six mice in each treatment group.

Measurement of infiltration of inflammatory cells Infiltration of inflammatory cells was determined by the same image analysis system using the section taken from the mouse skin block embedded in paraffin or Spurr resin and stained with hematoxylin and eosin or toluidine blue respectively. For each section, 10 tables were randomly chosen, unless otherwise indicated, and cell density was determined per mm2 of field. The average value for the 10 fields was obtained and entered as a single value for each mouse. The mean and SEM were calculated for the six mice in each treatment group. The measurement of inflammatory cell infiltration included background values that included other materials in the skin stained in the same manner. However, the increase, if it occurred, reflected the migration of inflammatory cells.

Treatment of data The percent inhibition of the drug on skin responses induced by TPA (ie, epidermal hyperplasia, and infiltrations of inflammatory cells and skin fold thickness) was calculated using the following formula: Total response due to TPA - Total response due to the drug% Inhibition = x! 00% Total response due to TPA - Total response due to acetone It is commonly found that the relationship between dose (or concentration) and the response can be described satisfactorily using the Michaelis-Menten equation or a variant thereof such as the Hill equation. This provides estimates of (a) the power, (b) the efficiency or maximum effect (Ema?) And the slope of the logarithm curve of concentration-response using the Hill coefficient (?). The Hill equation can be expressed as follows: Emax x C? E = CI50Y + C? Where IC50 is the concentration of drug that produces 50% of the maximum response and Emax refers to the maximum effect produced by the drug and is also referred to as efficacy. Efficacy is the measure of the intrinsic ability of a drug to initiate a response once it takes up the receptor sites. Measurements of both Emax and Cl50 (potency) are clearly crucial when compared to the activity of similar drugs. The Hill coefficient (?) Measures the slope of the dose-effect curve which can be markedly influenced by the shape of the curve that describes the binding of the drug to the receptor. For many drugs,? remains between 0.6 and 1.5. the use of Hill coefficient not only improves the fit of data but also indicates the influence of changes in dose or concentration on the response: for example, when? is greater than 1, the slope is very steep, meaning that a marked change in the drug effect is associated with a small change in the dose or concentration of the drug. On the other hand, when? is less than 1, with a low hyperbolic effect-concentration ratio, the activity occurs over a wide range of drug levels. Therefore, the different values of? they can dramatically affect the clinical usefulness of the drug. In the present study, the concentration-response curves were obtained for a series of corticosteroids. In these experiments, the adjustment of the curve was achieved using a computer program called "The Scientist" (MicroMath Scientific Software, Salt Lake City, Utah USA) in which the parameters Emax, Cl50 and Hill coefficient (?) For Each spheroid was estimated using non-linear regression and least-squares adjustment. The overall significance of the differences between treatments was determined by analysis of variance, while the Tukey HSD test was used to examine the level of significance of specific contrasts. The program for Windows Systat (Systat Inc., Evanston, Illinois, USA.) Was used.

EXAMPLE 2 Effects of TPA on mouse skin This study was conducted to determine the time course of effects produced by the application of TPA to mice, which were then sacrificed at intervals of 1, 2, 3, 5 and 8 days. The peak times for epidermal hyperplasia, dermal inflammation and skin fold thickness were determined. The purpose of this study was to determine the best time to sacrifice TPA-treated mice so that a maximum response to TPA could be obtained. The literature indicates that epidermal hyperplasia and dermal inflammation induced with TPA reach peaks at different times. It was expected, therefore, that a commitment time would have to be chosen. Forty-four female mice of the BALB / c strain were divided into treatment groups and the mice were treated with a single application (100μL) of TPA (0.01% in acetone) and sacrificed on days 1, 2, 3, 5 and 8 and the effects of the time course of the effects of TPA on epidermal hyperplasia, infiltration of inflammatory cells to the dermis and skin fold thickness as described in example 1 were measured. TPA are summarized as follows: Day l Day 2 Day 3 Day 5 Day 8 Increase in the thickness of the epidermis 326% 382% 393% 270% 130% Increase in the density of the dermis 397% 296% 272% 280% 225% Increase in the thickness of skin folds 193% 142% 124% 1 16% 107% The experiment showed that an individual application of TPA caused epidermal thickening, dermal inflammation and an increase in skin fold thickness that lasted for at least 8 days. Peak effects were observed at 72 hours for epidermal hyperplasia and 24 hours for dermal inflammation and skin fold thickness. Based on these results it was decided that an appropriate commitment time, for most experiments, would be to sacrifice the animals 24 hours after an individual application of TPA. This would result in maximum effects for inflammation and thickening of the skin fold and effects close to the maximum for epidermal hyperplasia (about 80% peak effect). Unless otherwise indicated, the mice were sacrificed 24 hours after the application of TPA.

EXAMPLE 3 Action of the different corticosteroids The ability of corticosteroids themselves to inhibit TPA lesions was first investigated. The following corticosteroid groups were tested (the classification of corticosteroid potencies is dependent on the concentration used, the composition of the vehicle and the effect being studied) (quadral) Low potency: betamethasone, dexamethasone, hydrocortisone, hydrocortisone acetate. Medium potency: alclometasone dipropionate, fluocortolone pivalate 21. High test: betamethasone dipropionate, betamethasone valerate, fluocinolone acetonide, halcinonide, mometasone furoate and triamcinolone acetonide. The mice were divided into treatment groups. Corticosteroids and TPA were premixed at the required concentrations and applied immediately to the backs of the mice. For those spheroids not very soluble in acetone (ie, betamethasone, dexamethasone, hydrocortisone and hydrocortisone acetate), the drugs were dissolved in 100 μL of dimethylformamide before further dilution with acetone. The efficacy of corticosteroids to inhibit epidermal hyperplasia, infiltration of inflammatory cells and thickening of skin fold as described above was evaluated. The concentration-response curves were determined after adjusting the data by computer. The curve fitting technique was generated by a computer program called "The Scientist" in which the parameters of the Hill equation were generated. These included EmaX? Cl50 and the Hill coefficient (?) For each steroid. The concentration-response curves for each steroid were plotted and the relative potencies were determined from the Cl 50 values. Comparisons were made of the variations in the Cl50 ratios for each corticosteroid tested. These values were compared with those found in the literature and reflected the relative intrinsic potencies of the spheroids.

TABLE 2 A summary of the concentration-response curves for the inhibition of epidermal hyperplasia induced by TPA by several spheroids showing the values of Ema ?, IC50 and gamma (?) With respect to the Hill equation.

The results are represented as the mean ± SD. The numbers in parentheses are the confidence intervals.

TABLE 3 A summary of the concentration-response curves for the inhibition of inflammatory cell infiltration induced by TPA-by several spheroids showing the values of Ema ?, CI50 and gamma (?) With respect to the Hill equation Emax (% of Corticosteroids IC50 (M x 104) Gamma (?) Inhibition) Dipropionate of 82.12 +.5.65 0.32 + 0.09 0.97 + 0.26 alclometasone (67.58-96.66) (0.10-0.54) (0.31-1.63) Betametasona 74.55 +.3.92 13.12 + 1.65 1.28 + 0.17 (66.09-83.01 (8.87 + 17.37) (0.85-1.71) Dipropionate 90.77 +.3.71 0.29 + 0.06 0.52 + 0.66 Betametasone (83.18-98.37) (0.16-0.41) (0.39-0.65) Valerate 73.52 + 1.76 0.29 + 0.03 0.93 + 0.07 betamethasone (69.46-77.57) (0.22-0.34) (0.77-1.09) Dexamethasone 99.05 + 10.98 4.44 + 1.02 0.73 + 0.19 (72.18-125.93) (3.19-5.69) (0.25-1.20) Acetonide of 79.60 + 2.41 0.16 + 0.02 1.05 + 0.14 fluocinolone (74.05-85.15 (0.11-0.21 (0.72-1.38) Pivalate 21 from 97.91 + 12.16 1.10 + 0.40 0.94 + 0.21 fluorocortolone (64.15 +131.67) (0.55-1.65) (0.36-1.54) Halcinonide 90.34 + 6.29 0.31 + 0.05 0.60 + 0.11 (74.94-105.74) (0.14-0.48) (0.32-0.88) Hydrocortisone 100.29 + 9.36 61.48. + 9.66 0.57 + 0.26 (71.35-129.23) (39.13-83.83) (0.25-089) ) Acetate of 79.36 + 11.49 77.33 + 13.85 1.11 + 0.27 hydrocortisone (59.39-99.33) (60.28-94.58) (0.42-1.80) Furoate of 89.98 + 4.88 0.20 + 0.05 0.69 + 0.14 mometasone (78.45-101.51) (0.08-0.33) (0.37-1.01) Acetonide 77.24 + 1.26 0.39 + 0.03 1.42 + 0.11 triamcinolone (74.15-80.33 0.33-0.46) (1.14-1.70) The results are represented as averages + SD. The numbers in parentheses are the confidence intervals. Tables 2 and 3 provide an estimate of the efficiencies (Emax), potencies (Cl50) and the slope of the concentration-response curve (?) For the 12 corticosteroids investigated in this study. These values were obtained from the Hill equation which is a modified form of the Michaelis-Menten equation. Apparently excellent correlations between Cl50 values could indicate that the inhibition of epidermal hyperplasia and inflammation are mediated by the same or some other mechanism, that the limiting factor in the production of the two effects was the ability of the steroid to penetrate the skin. The fact that the maximum effects (Ema?) And slopes (?) Of the concentration-response curves for the two effects were correlated very poorly suggests that different mechanisms are involved in the suppression of inflammation and epidermal hyperpiasia by the spheroids. In addition, correlations between IC50 values are not as impressive when the effects of certain drugs other than those studied are withdrawn. The effect of removing these different drugs is shown in Table 4: TABLE 4 Clsn values (inflammation vs epidermal hyperplasia) r; (read from the graph in Graph of the values of r IC50 (inflammation vs, (obtained from the equation where IC50 = concentration of epidermal hyperplasia) of Hill), inhibits half of the effect of TPA) All drugs included, 0.961 (p < 0.0005) 0.909 (p < 0.0005) Hydrocortisone and hydrocortisone acetate not 0.959 (p < 0.0005) 0.988 (p < 0.0005) included Hydrocortisone, hydrocortisone acetate, betamethasone dipropionate, 0.741 (p = 0.057) 0.637 (p = 0.072) dexamethasone and fluorocortolone pivalate 21 including 1 í From the above results it seems likely that the spheroids inhibit the inflammatory and hyperphasic effects of TPA by different mechanisms, either by inducing different biochemical responses or by producing the same response in different cell lines. This conclusion is quite relevant for the possible synergistic effects of auranofin that are described. The determination of the values of Ema? from the Hill equation could be subject to error due to some uncertainty about the measurements made at the top of the concentration-response curves. Thus, the Cl50 values were determined by two methods: (a) using the value Ema? generated from the Hill equation (tables 5 and 6) and (b) directly from the concentration-response curve, taking the CI5o value as the concentration that inhibited 50% of the inflammation and hyperplasia induced by TPA (tables 7 and 8). The difference between the IC50 values, and therefore the relative powers, determined by the two methods was not so great. However, the direct reading of the concentration-response curves was considered as more reliable and these readings were used to calculate the "synergistic factors" given in tables 9 and 10.

TABLE 5 Normal and relative potencies of typical corticosteroids for the inhibition of epidermal hyperplasia induced by TPA [values generated from the Hill equation where IC50 is the concentration that produced 50% of the maximum inhibitory effect (Emax)].

TABLE 6 Normal and relative potencies of topical corticosteroids for the inhibition of inflammatory cell infiltration induced by TPA in the dermis [values generated from the Hill equation where Cl50 is the concentration that produced 50% of the maximum inhibitory effect (Ema?) .

IC50 for inhibition of potency Corticosteroids infiltration of inflammatory relative cells in the dermis Hydrocortisone acetate 77.33 1.0 Hydrocortisone 61.48 I 1.26 Betametasona | 13.12 '5.89 Dexametasona 4.44! 17.42 Fluocortolone Pivalate 21 1.10 70.30 Triamcinolone acetonide 0.39 198.28 Alclometasone dipropionate 0.32, 241.66 Halcinonide: 0.31! 249.45 Betamethasone dipropionate 0.29! 266.66 Betametasone valerate 0.29 '266.66 Mometasone Furoate 0.20 1 386.65 Fluocinolone acetonide 0.16 483.31 TABLE 7 Normal and relative potencies of topical corticosteroids for the inhibition of epidermal hyperpiasia induced by TPA (values taken from the concentration-response curve where IC50 is the concentration that inhibited 50% of the effect of TPA) TABLE 8 Normal and relative potencies of topical corticosteroids for the inhibition of inflammatory cell infiltration induced by TPA in the dermis (values taken from the concentration-response curve where Cl50 is the concentration that inhibited 50% of the effect of TPA) In the present study, hydrocortisone, hydrocortisone acetate, betamethasone and dexamethasone had low to medium potencies with respect to both the inhibition of epidermal hyperplasia induced by TPA and the infiltration of inflammatory cells induced by TPA. This hierarchy was consistent with the correlation powers in the clinical field that was demonstrated in table 1. It was found that halcinonide, triamcinolone acetonide, alclometasone dipropionate, betamethasone dipropionate and fluocortolone pivalate 21, have medium potencies. to strong, in the inhibition of epidermal hyperplasia induced by TPA and the infiltration of inflammatory cells induced by TPA, and these values were consistent with the clinical potencies. Finally, mometasone furoate, betamethasone valerate, betamethasone dipropionate, and fluocinolone acetonide had strong to very strong potencies for epidermal hyperplasia induced by TPA. Among them, betamethasone valerate proved to be the most potent agent. When their potencies for the inhibition of inflammatory cell infiltration induced by TPA were investigated, they also proved to have a strong to very strong anti-inflammatory effect. Fluocinolone acetonide was found to be the most potent agent, with a relative potency of 483 (Table 6) or 440 (Table 8).

When comparisons were made between the relative potencies of the steroids tested in this study and their respective clinical potencies, the order of potency was generally the same. It could be expected that a particular corticosteroid could have equal potency with respect to anti-hyper- and anti-inflammatory actions. The results show that these reactions are not necessarily closely related. The linear regression of the relative potencies for the inhibition of the hyperplastic effects of the steroids against the relative potencies for the suppression of inflammation gave a value r = 0.573 (p = 0.51) for the data from tables 5 and 6 and a value of r = 0.658 (p <0.02) for the values in tables 7 and 8. In this way, about 65% of the variance between the two actions seems to be due to some common property (which could be the solubility in lipids) but a significant component of these actions differs with respect to the suppression of hyperplasia and the suppression of inflammation. This is also illustrated by comparing the orders of power hierarchy in tables 7 and 8. Only 6 of the 12 spheroids have the same hierarchy for both effects and, of these spheroids four are the four least powerful.

EXAMPLE 2 Synergistic effect of auranofin and corticosteroids with different clinical potencies The results of preliminary studies indicated that auranofin, under certain conditions, may inhibit some of the effects of TPA, although it is not particularly potent in this regard. The present study examines whether or not combinations of auranofin and corticosteroids have synergistic effect to suppress TPA lesions.

Four costicosteroids with different clinical potencies ranging from weak to strong were chosen, namely hydrocortisone, fluocinolone acetonide, betamethasone dipropionate and mometasone furoate. The TPA was premixed with the corticosteroids in the presence or absence of a fixed concentration of auranofin (0.2%) to produce the required concentrations and was applied to the backs of the mice immediately. The concentration-response curves were determined by non-linear regression using the least squares fit. The Cl50 values were determined by reading the value of the graph that corresponded to 50% of the effect produced by TPA (tables 9 and 10).

The term "apparent Cl50" refers to the value for the combination of steroid and auranofin.

TABLE 9 Apparent CI50 values obtained from the computer-adjusted graph for the effects on epidermal hyperplasia induced by TPA of four corticosteroids in the absence and presence of auranofin (0.2%).

The results are presented as the mean ± SD.

TABLE 10 Apparent CI50 values obtained from the computer-adjusted graph for the effects on the infiltration of dermal inflammatory cells induced by TPA of four corticosteroids in the absence and presence of auranofin (0.2%).

The results are presented as the mean ± SD.

The value called the "synergistic factor" is defined as the Cl50 value for the steroid determined in the absence of auranofine divided by the IC50 for the same steroid determined in the presence of auranofin (0.2%).

Synergism refers to situations in which a combination of two drugs produces an effect that is significantly greater than the algebraic sum of the effects when the same dose or concentration of each drug is observed separately in the same test system. Synergism may result in multiple potentiation of the effects of one or both drugs or this may give rise to effects that are qualitatively different from those shown by drugs when used separately.

With regard to the effects on epidermal hyperplasia, only two of the four steroids tested could be considered as showing a synergistic reaction with auranofin, namely betamethasone dipropionate and fluocinolone acetonide. From the results shown in tables 9 and 10 it can be seen that in the case of betamethasone dipropionate and fluosinolone acetonide, the percentage increase in the apparent potencies of steroids is 5.600% and 1.3%, respectively . However, in the case of the less potent of the four steroids, namely hydrocortisone, the apparent increase in potency was 100%. A more effective way to demonstrate the presence of true synergism is to separately compare the following: 1) auranofin (0.2%) alone, 2) a low steroid concentration only (enough to inhibit about 20% of the effects of TPA - this value can be read from the concentration-response curves for the steroid when studied separately), 3) the same concentration of steroid as in part (2) combined with auranofin (0.2%) -this value can be obtained from the dose response curve for the steroid in the presence of auranofin and 4) a concentration of the same steroid used in subsection (2) but in sufficient concentration to produce the same effect as that obtained in subsection (3). If the effects of subsection (3) appear to be the sum of clauses (1) and (2), the result is not synergism. If the effects of subsection (3) far exceed those of subsections (1) and (2) if they are added arithmetically, the result is synergism according to our definition.

The results for epidermal hyperplasia are shown in Figure 1. Aurofine alone inhibited epidermal hyperplasia induced by TPA by about 10% in all four studies. Betamethasone dipropionate 1x10 5 M alone caused about 20% inhibition of the effect of TPA The combination of betamethasone dipropionate (1x10 5 M) and auranofin (0.2%) produced about 65% inhibition of TPA effect . To gauge the significance of the increased effect that resulted when auranofin was added to betamethasone dipropionate, a comparison should be made with the concentration of betamethasone dipropionate which, in the absence of auranofin, produced the same effect. This value was 5x10"4 M or 50 times the concentration that produced the same effect in the presence of auranofin.

With respect to epidermal hyperplasia, studies showed that: a) massive synergism results when auranofin is added to betamethasone dipropionate and fluocinolone acetonide; b) a lesser degree of synergism may result from the combination of hydrocortisone and auranofin; and c) that there is no synergism or even additive effects when the auranofin is coadministered with mometsone furoate.

With respect to the infiltration of inflammatory cells in the dermis, the results in Table 10 indicate that the effects of a combination of auranofin (0.2%) and mometasone furoate is the result of synergism since the apparent CI5o increases by about 3,800%. It is also possible that a lesser degree of synergism occurs with respect to the anti-inflammatory action of betamethasone dipropionate in the presence of 0.2% auranofin. Here the increase in apparent Cl50 is in the order of 400-500% (Table 10).

The results in figure 2 show a massive synergism between mometasone furoate (5x10"6 M) and auranofin (0.2%). This combination produced an effect that was equal to that produced by mometasone 1x10"3 M in the absence of auranofin Figure 2 also indicates that synergism could exist for the combination of auranofone with betamethasone dipropionate and fluocinolone acetonide, but not with hydrocortisone.

The results of these studies indicate that the extensive synergism results from the combination of auranofin with certain corticosteroids, such as for example betamethasone dipropionate and fluocinolone acetonide in consideration of the reduction of epidermal hyperplasia, and with others such as mometasone furoate for example. , in consideration of the reduction of inflammation. There could be less degrees of synergism between auranofin and other steroids.

Gold compounds and corticosteroids, as well as their formulations, which can be appropriately used in the present invention have been discussed in detail in the present application or Australian patent number 616 755, which is incorporated herein by reference .

Claims (37)

NOVELTY OF THE INVENTION CLAIMS

1. The use of a gold compound in combination with a corticosteroid for the manufacture of a medicament for the treatment of an immune-mediated disorder having an inflammatory component and a cellular hyperproliferation component or both, characterized in that at least one corticosteroid is selected for interacting in the gold compound so that it exhibits preferential synergistic action towards one of the components of said disorder or that exhibits an equal action towards each component of said disorder.

2. The use according to claim 1, further characterized in that the disorder has an inflammatory component and a cellular hyperproliferation component.

3. The use according to any of the preceding claims further characterized in that the gold compound and at least one corticosteroid are administered simultaneously.

4. The use according to any of the preceding claims, further characterized in that the gold compound and at least one corticosteroid are administered sequentially.

5. The use according to claim 4, further characterized in that at least one corticosteroid is administered after the gold compound.

6. The use according to any of the preceding claims further characterized in that at least two corticosteroids are administered, of which at least one is selected to interact with the gold compound to exhibit preferential synergistic action towards the inflammatory component, and at least the other is selected to interact with the gold compound so that it exhibits preferential synergistic action towards the cellular hyperproliferation component of said disorder.

7. The use according to any of the preceding claims, characterized in that the disorder is an immune-mediated dermatological disorder.

8. The use according to claim 7, characterized in that the disorder is psoriaris.

9. The use according to claim 7, further characterized in that the disorder is dermatitis.

10. The use according to any of claims 1 to 6, further characterized in that the disorder is rheumatoid arthritis.

11. The use according to any of the preceding claims, further characterized in that the gold compound is fat-soluble.

12. The use according to any of the preceding claims, further characterized in that at least one corticosteroid is selected to interact with the gold compound to exhibit synergistic action toward cell hyperproliferation in preference to inflammation.

13. The use according to claim 12, further characterized in that at least one corticosteroid is selected from the group consisting of betamethasone dipropionate, fluocinolone acetonide and hydrocortisone.

14. The use according to any of the preceding claims, further characterized in that at least one corticosteroid is selected to interact with the gold compound to exhibit synergistic activity toward inflammation in preference to cellular hyperproliferation.

15. The use according to claim 14, further characterized in that at least one corticosteroid is selected from the group consisting of betamethasone dipropionate, fluocinoione acetonide and mometasone furoate.

16. The use according to claim 10, further characterized in that the corticosteroid is selected from the group consisting of hydrocortisone acetate, hydrocortisone, betamethasone, betamethasone dipropionate, dexamethasone, fluocortolone pivalate 21, triamcinolone acetonide, valerate betamethasone, alclomethasone dipropionate, halcinonide, mometasone furoate and fluocinolone acetonide.

17. - The use according to claim 16, further characterized in that the corticosteroid is selected from the group comprising hydrocortisone, betamethasone dipropionate, mometasone furoate and fluocinolone acetonide.

18. The use according to any of the preceding claims, further characterized in that the gold compound is auranofin.

19. The use according to any of the preceding claims, further characterized in that the gold compound is administered systemically.

20. The use according to any of claims 1 to 18, further characterized in that the gold compound is administered orally.

21. The use according to any of claims 18 to 18, further characterized in that the gold compound is administered locally.

22. The use according to any of claims 1 to 18, characterized in that the gold compound is administered topically.

23. The use according to any of claims 18 to 18, further characterized in that the gold compound is administered by intra-articular injection.

24. - The use according to any of the preceding claims characterized in that at least one corticosteroid is administered systemically.

25. The use according to any of claims 23 to 23, further characterized in that at least one corticosteroid is administered orally.

26. The use according to any of claims 23 to 23, further characterized in that at least one corticosteroid is administered locally.

27. The use according to any of claims 23 to 23, further characterized in that at least one corticosteroid is administered topically.

28. The use according to any of claims 23 to 23, further characterized in that at least one corticosteroid is administered by intra-articular injection.

29. A pharmaceutical composition that includes a gold compound and one or more corticosteroids, the corticosteroids being selected to interact with the gold compound to exhibit a preferential synergistic action towards an inflammatory component, a cellular hyperproliferation component or both of a immune mediated disorder, in combination with a pharmaceutically acceptable vehicle, excipient, auxiliary or solvent.

30. - A pharmaceutical composition according to claim 29, further characterized in that the composition is formulated for systemic administration.

31. A pharmaceutical composition according to claim 29, further characterized in that the composition is formulated for oral administration.

32. A pharmaceutical composition according to claim 29, further characterized in that the composition is formulated for local administration.

33. A pharmaceutical composition according to claim 29, further characterized in that said composition is formulated for topical administration.

34.- A pharmaceutical composition according to claim 29, further characterized in that the composition is formulated for administration by intra-articular injection.

35.- A pharmaceutical composition according to any of claims 29 to 34, further characterized in that the corticosteroid is selected from the group comprising hydrocortisone acetate, hydrocostisone, betamethasone, betamethasone dipropionate, dexamethasone, fluocortolone pivalate 21, acetonide of triamcinolone, betamethasone valerate, alclometasone dipropionate, halcinonide, mometasone furoate and fluocinolone acetonide.

36. The composition according to claim 35, characterized in that the corticosteroid is selected from the group consisting of hydrocortisone, betamethasone dipropionate, mometasone furoate and fluocinolone acetonide. 37.- The composition according to any of claims 29 to 36, characterized further because the gold compound is auranofin.