IE62006B1 - "Orthorhombic ciclosporin crystal form, process for its production and pharmaceutical compositions containing it" - Google Patents

Abstract

Pharmaceutical compositions, in particular topical forms, e.g. for dermal or topical ophthalmic use, and injectible forms, e.g. intra-lesional or intra-articular injection, comprising Ciclosporin as active ingredient in orthorhombic crystal form. Ciclosporin in non-solvate orthorhombic crystal form is also described together with its use and processes for its production.

Description

Iq The present invention relates to novel pharmaceutical compositions comprising Ciclosporin as active ingredient.

Ciclosporin, or cyclosporin A, is a known fungal metabolite having valuable pharmaceutical utility. Ciclosporin is a cyclic poly-Nlg methylated endecapeptide of formula -MeBm t-aAbu-Sar-MeLeu-Val-MeLeu-Ala-(D)Ala-MeLeu-MeLeu-MeVal1 234567 8 9 10 11 in which -MeBmt- represents an N-methyl-(4R)-4-but-2E-en-l-yl-4methyl-(L)threonyl residue.

Ciclosporin as well as methods for its production are, for example, 2g described in US patent no. 4,117,118. The primary area of clinical investigation for ciclosporin has been as an immunosuppressive agent, in particular in relation to its application in treating recipients of organ transplants, e.g. heart, lung, combined heart-lung, liver, kidney, pancreatic, bone-marrow, skin and corneal transplants and, in 2Q particular, allogenic organ transplants. In this field Ciclosporin has » achieved a remarkable success and reputation and is now widely employed in clinic and commercially available under the Registered Trade Mark SANDIMMUN* or SANDIMMUNE8, for example in the form of an infusion concentrate, of an oral solution and of a gelatin encapsulated solution, the latter tvo variants being described and claimed for example in US patent no. 4,388,307.

At the same time, investigation of the applicability of Ciclosporin to various autoimmune diseases and to inflammatory conditions, in particular inflammatory conditions vith an aetiology including an autoimmune component such as arthritis (for example rheumatoid arthritis, arthritis chronica progrediente and arthritis deformans) and rheumatic diseases, has been intensive and reports and results in vitro, in animal models and in clinical trials are wide-spread in the literature. Specific auto-immune diseases for vhich Ciclosporin therapy has been proposed or applied include, autoimmune hematological disorder (including e.g. hemolytic anaemia, aplastic anaemia, pure red cell anaemia and idiopathic thrombocytopaenia), systemic lupus erythematosus, polychondritis, sclerodoma, Vegener granulamatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, psoriasis, Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease (including e.g. ulcerative colitis and Crohn's disease), pemphigus, endocrine opthalmopathy, Graves disease, sarcoidosis, multiple sclerosis, primary billiary cirrhosis, primary juvenile diabetes (diabetes mellitus type I), Behcet's disease, uveitis (anterior and posterior), vernal conjunctivitis, keratoconjunctlvlts sicca, interstitial lung fibrosis, psoriatic arthritis and glomerulonephritis (vith and without nephrotic syndrome, e.g. including idiopathic nephrotic syndrome or minimal change nephropathy).

Further areas of investigation have included potential applicability as an anti-parasitic, in particular anti-protozoal agent, vith possible uses suggested including treatment of malaria, coccidiomycosis and schistosomiasis, as well as use in reversing tumor resistance to chemotherapeutic, e.g. cytostatic, therapy and in promoting hair growth, e.g. for the treatment of alopecia.

In the solid state, Ciclosporin exists in both amorphous and crystalline form. In the crystalline state, both a tetragonal and an orthorhombic modification are knovn.

The tetragonal modification (P4X), also referred to herein for convenience as Cy-A/X-I, is described together vith processes for its production, e.g. in Austrian patent specification no. 353,961.

Cy-A/X-I has a latice a=b« 13.8 A c» 41.2k, volume per asymmetric unit = 1974 A3, a melting point at ca. 140-150 °C and comprises ca. 2 molecules H20 per Ciclosporin molecule.

A first orthorhombic modification (Ρ2ι2χ2ι) is obtainable by recrystallisation from di-isopropyl ether as discussed in an article entitled Total synthesis - change in molecular structure - biological effect: Ciclosporin as example by R. Wenger, in Sandorama 1984/ III. This particular modification comprises a Ciclosporin/di-isopropyl ether (ca. 1:2) solvate, vith a crystal lattice of a - 12.5 A, b = 22.9 A, c = 28.4 A, vol per asymmetric unit 2027 A3, and a melting point at ca. 150 °C. Orthorhombic Ciclosporin solvate crystals are obtainable by recrystallisation of supersaturated Ciclosporin solutions in an appropriate solvent medium over prolonged periods of time. Thus the orthorhombic Ciclosporin/di-isopropyl ether solvate modification prepared by Wenger is obtained by addition of a concentrated solution of Ciclosporin in methylene chloride (in vhich Ciclosporin is rather readily soluble) to di-isopropyl ether (in vhich Ciclosporin is relatively less soluble) and evaporation of the methylene chloride. Crystallisation from the remaining di-isopropyl ether is then alloved to proceed over a period of ca. 7 days at ambient temperature. Orthorhombic crystal forms comprising such Ciclosporin solvates are also referred to, for convenience, herein generically as CY-A/X-II.

* A further orthorhombic (P2i2i2x) modification has nov been identified, characterised by the absence of any solvent component. -, The dimensions of the crystal lattice for the solvent free or non-solvate orthorhombic Ciclosporin modification are a » 12.7 A, b = .7 A, c = 36.3 A, vol. per asymmetric unit = 1804 A3. The melting point is of the order of from about 180 to about 190 °C up to about 195 °C, e.g.--depending on degree of purity/source as indicated in Examples 1 and 2 hereinafter, in vhich specific methods for the obtentlon of this modification are described. Cidosporin in non-solvate (or solvent free) orthorhombic crystal form is also referred to herein for convenience as CY-A/X-III. Further characterising (X-ray povder diffraction) data for CY-A/X-III are provided in Table III and Fig. IV hereinafter, together vith corresponding data for CY-A/X-I and CY-A/X-II.

IQ Despite the very considerable success and reputation vhich Cidosporin has achieved and despite the major contribution to medical science, in particular to the art of organ transplant, vhich it has already made, its use in practice is still not ideal or problem free. A particular area of difficulty vhich has been encountered is in the provision of galenic forms permitting convenient or vholly satisfactory administration, or best suited for the treatment of specific diseases or conditions for vhich Cidosporin has already been identified as a potential drug of choice. 2Q These difficulties are in part attributable to the particular physical characteristics of Cidosporin, for example its relatively limited solubility or (hitherto experienced) compatibility vith available, pharmaceutically applicable carrier media. Such difficulties are compounded by the very nature of certain of the disease conditions for vhich Cidosporin represents potential first line therapy as veil as the occurrence of undesirable side effects vhich Cidosporin may induce vhen administered orally. For example Cidosporin has been identified as effective in the treatment of psoriasis folloving oral administration. Psoriasis is a vide-spread condition causing 3Q considerable patient suffering and for vhich there is at present no generally accepted, effective therapy. Sufferers from psoriasis clearly represent a patient population at need. Vhile available evidence vould suggest that Cidosporin might meet this need it is improbable that oral Cidosporin therapy could be justified as a con35 ventional means of psoriasis treatment. Similar considerations apply to the treatment of e.g. eye diseases such as posterior uveitis and keratoconjunctivitis sicca.

Various systems have been proposed for the administration of Ciclosporin, including galenic forms intended for topical or dermal application or for ophthalmic application, i.e. topical application to the eye, e.g. for possible use in the treatment of psoriasis, of other dermatological diseases and conditions, for example atopic dermatosis and alopecia, or for the treatment of uveitis. Such systems have however hitherto not brought any notable benefit, or have not proven wholly satisfactory.

Thus variously reported attempts to treat psoriatic subjects by the application of the knovn oral solution directly at the superficial site of psoriatic lesion have met with minimal or no success and success in relation to subjects suffering from atopic dermatosis has been strictly limited. Similarly, while topical application to the eye has been found to be widely effective as a means of preventing e.g. corneal transplant rejection, or of treating disease in the anterior segment of the eye, no topical formulation has hitherto been reported which might permit ready, safe or effective treatment of diseases or conditions as they affect sites deeper within the eye, for example for the general treatment of uveitis, including posterior uveitis.

Comparable difficulties have been encountered in relation to the development of other ciclosporin drug delivery systems, for example injectible forms, e.g. suitable for intra-articular injection for the local treatment of arthritic diseases, e.g. as hereinbefore set forth, or for intra-lesional injection, e.g. for the treatment of severe psoriatic lesion, or alternative oral delivery systems, e.g. having modified bioavailability or drug-release characteristics.

Hitherto attempts to formulate Ciclosporin have made use of the compound in solution or, less commonly, in amorphous or Cy-A/X-I form, the latter being, for example, the crystal form identified e.g. for drug registration purposes. And despite the very considerable efforts expended in seeking new or improved alternatives, the oral solution and infusion concentrate already referred to remain the only widely applied delivery systems for Ciclosporin.

In accordance with the present invention, it has now surprisingly been found that Cy-A/X-III, is of especial advantage and benefit in the preparation of ciclosporin galenic formulations of various and diverse kind.

In particular it has been found that such a crystal modification may be used to prepare galenic formulations comprising Ciclosporin in stable, fine particulate form and/or having enhanced stability or release characteristics. By this finding the way is opened to the production of novel and improved Ciclosporin drug delivery systems, e.g. possessing sustained or delayed Ciclosporin delivery characteristics and/or adapted to or suitable for the treatment of diseases or conditions hitherto refractory to Ciclosporin therapy or for which conventional Ciclosporin therapy has been inherently less suited. More especially, it has been found that by use of such a crystal modification, e.g. in fine particulate form as aforesaid, galenic forms may be obtained suitable for topical or dermal application or for topical ophthalmic application, for the treatment of diseases or conditions affecting the skin or eye, e.g. as hereinbefore set forth, in particular for the treatment of psoriasis, atopic dermatitis, contact dermatitis, occular symptoms of Behchet's disease, uveitis and keratoconjunctivitis sicca. Use of such a crystal modification also enables the preparation of injectible forms for Ciclosporin having sustained or delayed release characteristics and hence of particular utility, e.g. in the treatment of diseases or conditions for which required frequency of administration may otherwise impede utility, for example for the treatment of arthritic or similar disease of the joint by intra-articular injection or for the treatment of psoriasis or similar disease of the skin by intra-lesional injection.

For such purposes CY-A/X-III has been found to be inherently much better suited than other available forms, in particular amorphous Ciclosporin or CY-A/X-I, e.g. in terms of physico-chemical properties, especially stability, release characteristics, i.e. release of Ciclosporin from the crystal lattice, and susceptibility to the necessary procedures of galenic formulation. In a first embodiment the present invention accordingly provides: A CY-A/X-III, i.e. Ciclosporin in non-solvate orthorhombic crystal form; in particular: A1 CY-A/X-III having a crystal lattice a a = 12.7 A, b= 15.7 A, c = 36.3 A, volume per asymmetric unit 1804 A, and/or A2 CY-A/X-III exhibiting the X-ray powder diffraction characteristics herein set forth in Table III.

The present invention also provides: B A process for the production of CY-A/X-III vhich process comprises crystallising Ciclosporin from a CY-A/X-III bearing solvent medium and recovering CY-A/X-III thus obtained.

Suitable CY-A/X-III bearing media for use in the above process are, in particular, solvent media for Ciclosporin comprising higher molecular weight solvent components, e.g. having a molecular veight >200. Especially suitable are media comprising higher molecular veight ethers, in particular polymeric ethers, for example polyalkyl ethers, e.g. polyethylene-, and polypropylene-glycols. Examples of polymeric ethers vhich have been found to be appropriate are polyethylene and polypropylene glycols, e.g. having a molecular veight of >200; glycerin-polyethyleneglycol esters, e.g. glycerin-polyethyleneglycolricinoleates and glycerin-polyethyleneglycoloxystearates as, for - 8example, available under the Trade Marks Cremophor EL and Cremophor RH (c.f. Fiedler, Lexikon der Hilfstoffe 2nd edition, Vol. 1, p.p. 257 and 258); polyoxyethylene-sorbitan esters, for example as available under the Trade Mark Tween (c.f. Fiedler, 2, p.p. 972-975); polyoxyethylene esters, e.g. polyoxyethylene-stearinic acid esters, for example as available under the Trade Mark Myrj (c.f. Fiedler, 2, p.p. 636); and trans-esterification products of natural oil triglycerides and polyalkylene polyols, including the IQ trans-esterification products of maize oil, kernel oil, almond oil, ground nut oil, olive oil, palm oil and mixtures thereof vith polyethylene-glycols, in particular polyethylene-glycols having a molecular veight of from 200 to 800, for example as available under the Registered Trade Mark Labrafil (c.f. Fiedler, p. 539).

Ig Especially suitable are media comprising polyethylene-glycols having a molecular veight of ca. 200 to ca. 600, in particular of ca. 300 to ca. 400.

As will be appreciated, the CY-A/X-III bearing medium may include one 2q or more components in addition to those recited above, for example lover molecular veight alcohols and/or vater, as a means of modifying solvent/crystallisation characterisitics. A particularly suitable CY-A/X-III bearing medium has been found to be one comprising ca. 0.5 parts by volume ethanol, ca. 8.0 parts by volume polyethylene-glycol (300) and ca. 1.0 parts by'volume H20.

To carry out the process of the invention Ciclosporin is first dissolved, e.g. in amorphous or tetragonal form in the selected CY-A/X-III bearing medium at elevated temperature. The dissolution 3Q temperature will of course vary depending on the particular medium chosen but will generally be >40eC more especially >50eC. In the case of polyethylene-glycol based media the dissolution temperature vill generally be >70°C, e.g ca. 75-130eC. Suitably the obtained solution vill comprise >5X, preferably >101, e.g. up to about 40-60X, preferably up to a maximum of 201 by veight Ciclosporin based on the total veight of the solution. Crystallisation of Ciclosporin is then allowed to proceed vith cooling, suitably over a relatively prolonged period of time, e.g. of the order of 10-35 minutes or more depending on desired crystal growth.

If desired nucleation procedures may be commenced, e.g. by sonication or seeding. Where ethanol/polyethylene-glycol/H20 crystal bearing media, e.g. as described above, are employed it has, for example, been found that nucleation can be readily initiated at temperatures of ca. 60-90°C, e.g. ca. 65-85°C. by application of sonic vibration at ca. 20,000 c.p.c. As indicated in example 2 hereinafter, by continuing sonication in this manner throughout the course of crystallisation, micro crystals may be obtained of very fine and relatively constant particle size.

The rate of crystal growth and the size of crystals obtained will of course, depend on the particular medium chosen and the conditions under which crystallisation is performed, for example on whether or not sonication is applied and at what frequency. For use in accordance with the present invention, e.g. for the preparation of galenic formulations as hereinafter described, CY-A/X-III will preferably be employed in fine particulate form, e.g. having an average particle size of <200uM, e.g. of from 0.5 to 200yM. Where initially obtained crystals are of relatively large size, particulate preparations having the desired size characteristics can, of course, be obtained by conventional comminution procedures, e.g. by grinding, milling, pulverising, micronising or otherwise fragmenting to reduce size.

While particulate preparations obtained in this way may be entirely adequate for some purposes, for example in the preparation of topical compositions for application to the skin, use of grinding or milling or equivalent techniqes will in many cases not be preferred. Reasons for this are that the products of such procedures tend to exhibit relatively wide particle size distribution and to have a relatively coarse or abrasive structure as a consequence of crystal fracture. In addition comminution will tend to destroy physical characteristics at the natural crystal surface and is in any event difficult or costly to perform while maintaining required conditions of sterility, e.g. in the manufacture of galenic formulations for intra-articular - 10 administration. For such reasons it is commonly preferred to carry out the method of the invention in such a manner as to obtain crystals, e.g. microcrystals, of dimensions vithin the desired end particle size range. Thus for use in the manufacture of e.g. compositions for intra-articular or other parenteral application use of CY-A/X-III particle preparations in non-comminuted microcrystalline state (i.e. CY-A/X-III microcrystals in native or non-fractured condition, e.g. vhich have not been subjected to grinding, milling, mlcronization or other reductive procedures) vill be preferred.

In order to obtain CY-A/X-III microcrystals vhich can be used directly in non-comminuted state in the above described manner, the process of the invention vill suitably be carried out vith agitation, for example by stirring, e.g. at 200-300 r.p.m. rising to e.g. 700 r.p.m. during crystallisation, and, especially, by application of ultra-sonic vibration, e.g. vithin the range of 10,000-30,000 cycles per sec, suitably of the order of about 20,000 cycles per sec.

By variation of the CY-A/X-III bearing medium and adjustment of the rate of cooling and degree of agitation during the recrystallisation procedures in accordance vith techniques knovn in the art and as hereinafter exemplified, CY-A/X-III microcrystals may be produced of varying average particle size, e.g. as set forth above or as hereinafter described for use in relation to individual types of galenic formulation in accordance vith the invention.

As previously indicated crystallisation in accordance vith the process of the invention vill suitably be performed under sterile conditions. Purification of the obtained CY-A/X-III may be carried out in conventional manner, e.g. by rinsing vith poly ethylene glycol (300) : vater e.g. in a ratio of 3-4:1 p.p.v./ vashing vith varm vater, as described in the examples. The obtained CY-A/X-III vill suitably be recovered free or substantially free of Ciclosporin in any other form, preferably in pure or substantially pure form. The rinsing step above assists prevention of the formation of amorphous Ciclosporin.

In accordance vith the foregoing the present invention additionally provides: CY-A/X-III producible by any of the above described procedures as veil as: A3 CY-A/X-III according to any one of A to A2 above, free or substantially free of Ciclosporin in any other form, e.g. free or substantially free of Ciclosporin in amorphous form, of CY-A/X-I or of CY-A/X-II; A4 CY-A/X-III according to any one of A to A3 above in pure or substantially pure form; A5 CY-A/X-III according to any one of A to A4 above in fine particulate form, vherein the component particles have an average particle size of <200yM, e.g. having particle size characteristics as hereinbefore set forth or as hereinafter described, e.g. in relation to galenic forms comprising CY-A/X-III; A6 CY-A/X-III according to any one of A to A5 above consisting predominantly, or consisting entirely or substantially entirely, of non-comminuted crystal material, e.g. CY-A/X-III according to A5 above vherein the component particles are comprised predominantly, or comprised entirely or substantially entirely, of non-comminuted microcrystals; and A7 CY-A/X-III according to any one of A to A6 above in sterile or substantially sterile condition, e.g. in a condition suitable for pharmaceutical use, in particular suitable for parenteral application, e.g. for administration by parenteral injection, e.g. intra-articular injection.

In addition the present invention also provides: C A pharmaceutical composition comprising a) Ciclosporin in non-solvate orthorhombic crystal form as active ingredient, together vith - 12 Ciclosporin is present in the compositions of the invention as CY-A/X-III, e.g. according to any one of A3 to A7 above.

Though Ciclosporin is present in the compositions of the invention in solid (crystalline) form, the compositions themselves may be of any appropriate constitution. Compositions in accordance with the invention thus include solid forms, such as powders and granulates, semi-solid forms such as ointments, gels, creams and pastes, as well as liquid forms, e.g. comprising the defined Ciclosporin component in fine particulate form (for example micronised or microparticulate form) suspended or dispersed in a liquid pharmaceutical diluent or carrier, in which the crystal form may be retained.

Compositions in accordance with the invention include both systems comprising a single Cidosporin-containing phase or compartment, for example simple creams, gels, ointments etc... as aforesaid or simple one compartment ampoules, for example ampoules for injection, as well as systems comprising a plurality of phases or compartments. Thus the invention is to be understood as embracing, e.g. compositions comprising a Cidosporin-containing phase or compartment together with one or more non-Ciclosporin-containing phases or compartments, e.g. coating, carrier, retard or placebo phases, or phases or compartments comprising other drug substances or adjuvant materials. The invention is thus to be understood as embracing, for example: 13mixed granulates comprising a first granulate phase comprising Cidosporin in orthorhombic crystal form and a second non-Ciclosporin-containing granulate phase; and split capsules having two or more compartments, e.g. divided by an internal vail, one of vhich contains Cidosporin in orthorhombic form and one of vhich does not. Other alternatives and variants vill be appparent to those skilled in the art.

Preferably Cidosporin vill be present on the compositions of the invention predominantly or, preferably, entirely or substantially entirely, as CY-A/X-III.

The compositions of the invention vill suitably comprise at least one phase or compartment comprising Cidosporin CY-A/X-III, throughout vhich said crystal form is distributed or distributable uniformly.

Such systems include for example homogeneous creams, ointments, gels and the like as veil as liquid systems in vhich the contained crystalline Cidosporin is or can be readily distributed homogenously, for example by hand agitation.

Compositions in accordance vith the invention may include: forms suitable for topical administration e.g. dermal or topical ophthalmic administration; for parentaral administration, e.g. by infusion or injection, including sub-cutaneous or intramuscular injection a.nd, in particular, intra-lesional or intra-articular injection; as veil as forms for enteral administration, for example suppositories, pessaries and the like. Enteral forms vill hovever generally be less preferred.

Preferably CY-A/X-III, vill be present in the compositions of the invention in fine particulate form, e.g. micronised, microparticulate or non-comminuted microcrystalline form as hereinbefore described. Appropriate particle size vill of course vary depending on the particular nature of the composition, its intended use and mode of application and the effect desired. In general hovever, average particle size vill suitably be of-- 14 the order of from 0.5-200μΜ. Particular ranges, e.g. in relation to the preparation of oral, topical or injectible forms are further discussed hereinafter.

TOPICAL FORMS e.g. for dermal or topical ophthalmic administration, e.g. for the treatment of psoriasis or atopic dermatosis or of diseases or conditions of the eye requiring immunosuppressive therapy, for example uveitis, conjunctivitis, including keratoconjunctivlts sicca or vernal keratoconjunctivitis, or corneal transplant.

Gels, creams, ointments and the like for topical or dermal administration can be prepared in accordance vith any of the techniques known and generally employed in the art, for example by compounding Ciclosporin in orthorhombic crystal form vith an appropriate flovable 15 base material, e.g. aqueous gel, for example aqueous cellulose-ether based gel, vith the addition of appropriate tensides, thickeners etc. including e.g. aerosil.

The Ciclosporin component is preferably present in fine particulate form, e.g. having an average particle size of from 0.5-200μΜ.

Preferably the average particle size is <60-80μΜ, more preferably <40μΜ, yet more preferably <30μΜ, most preferably <20μΜ.

Advantageously the particles vill have as narrov a particle size distribution as possible. Preferably substantially all particles present vill be less than 60-80μΜ. f Compositions of the invention for topical ophthalmic use, i.e. application to the eye, may include, for example suitably thickened suspensions or dispersions presented e.g. in the form of viscous eye-drop preparations or creams or gels for occular application. In the case of compositions for administration to the eye, the average particle size for the crystalline Ciclosporin material present vill suitably be <25μΜ. More preferably substantially all particles present vill be less than 25μΜ. Average particle size for such eye drop preparations vill suitably be in the range of from about 0.1, or preferably from about 0.5, to about 20μΜ or, preferably, to about ΙΟμΜ.

The hydrophilic/lipophilic content of the carrier system for use in forms for dermal application or for use in preparations for application to the eye may vary, e.g. depending on the particular surface to which the composition is to be applied, e.g. depending on the characteristics of the particular skin area to be treated.

Compositions for topical, in particular dermal, application suitably comprise a carrier medium in which the active ingredient (Ciclosporin component) is only slightly soluble. Suitably the solubility of the CY-A/X-III in such compositions is <10X, preferably <1.02 by weight. More preferably solubility is <0.12, most preferably <0.0012 by weight.

Suitable carrier media include hydrophilic, water miscible carriers as well as hydrophobic carriers.

Hydrophilic, water miscible carrier components are suitably of relatively low volatility. Examples of appropriate carrier components belonging to this class include liquid polyalkylene glycols, propylene glycol, glycerol and ethylene glycol. Suitable polyalkylene glycols include in particular, liquid polyethylene glycols, e.g. having an average molecular weight up to about 600, e.g. having an average molecular weight from about 250 to 450, especially ca. 300.

Representative compositions in accordance with the invention, e.g. for topical application, accordingly comprise: C1 a) Ciclosporin in non-solvate orthorhombic crystal form, i.e.

Cy-A/X-III, e.g* as defined under any one of A3 to A7 above; b) A hydrophilic, water miscible carrier, e.g. as described above; and, optionally c) Pharmaceutical grade water.

The amount of component (a) present vill suitably be of the order of from about 0.1 to about 30.OX, especially about 0.5 or 1.0 to 20X, e.g. ca. 1.0, 2.0, 5.0 or 10.OX based on the total veight of the composition. Components (b) and (c) vill suitably be present in a ratio of from 1:0 to 1:20 p.p.v. More suitably components (b) and (c) vill be present in a ratio of from 1:0.5 to 1:5, e.g. from 1:1 to 1:3, most suitably 1:1.5 to 1:2.5 p.p.v.

Such compositions vill suitably additionally comprise: d) A surface active agent.

Suitable components (d) include, inter al. d1 Trans-esterification products of natural vegetable oil triglycerides and polyalkylene polyols.

Such trans-esterification products are knovn from the art and may be obtained e.g. in accordance vith the general procedures described in US Patent No. 3 288 824. They include 2Q trans-esterification products of various natural (e.g. non-hydrogenated) vegetable oils for example, maize oil, kernel oil, almond oil, ground nut oil, olive oil and palm oil and mixtures thereof vith polyethylene glycols, in particular polyethylene glycols having an average molecular veight of from 2g 200 to 800. Preferred are products obtained by trans-esterification of 2 molar parts of a natural vegetable oil triglyceride vith one molar part of polyethylene glycol (e.g. having an average molecular veight of from 200 to 800). Various forms of trans-esterification product of the class defined are 3Q knovn and commercially available under the Trade Mark Labrafil [see Fiedler, Lexikon der Hilfstoffe, 2nd. revised and expanded edition (1981), Vol. 2, page 539]. Especially useful as components , of the compositions of the invention are the products: Labrafil M 1944 CS, a trans-esterification product of kernel oil and polyethylene glycol having an acid number of ca. 2, a saponification number of 145- 175 and an iodine number of 60 - 90; - 17 and Labrafil M 2130 CS, a trans-esterification product of a Ci2to C18- glyceride and polyethylene glycol having a melting point of ca. 35 - 40°C., an acid number of <2, a saponification value of ca. 185 - 200 and an iodine number of <3. d2 Reaction products of a natural or hydrogenated castor oil and ethylene oxide. Such products may be obtained in knovn manner, e.g. by reaction of a natural or hydrogenated castor oil vith ethylene oxide, e.g. in a molar ratio of from about 1:35 to about 1:60, vith optional removal of the polyethyleneglycol component from the product, e.g. in accordance vith the methods disclosed in German Auslegeschriften 1,182,388 and 1,518,819. Especially suitable are the various tensides available under the Trade Mark Cremophor. Particularly suitable are the products Cremophor RH 40 having a saponification number of ca. 50-60, an acid number <1, an iodine number <1, a vater content (Fischer) <2Z, an nD60 of ca. 1,453 - 1,4578 and an HLB of ca. 14 - 16; Cremophor RH60 having a saponification number of ca. 40 - 50, an acid number <1, an iodine number <1, a vater content (Fischer) 4.5-5.5X, an nD29 of ca. 1,453 - 1,57 and an HLB of ca. 15 - 17; and Cremophore EL having a molecular veight (by steam osmometry) of ca. 1630, a saponification number of ca. 65 - 70, an acid number of ca. 2, an iodine number ca. 28 - 3? and an nd25 of ca. 1,471. Also suitable for use in this category are the various tensides available under the Trade Mark Nikkol, e.g. Nikkol HC0-60. The said product Nikkol HC0-60 is a reaction product of hydrogenated castor oil and ethylene oxide exhibiting the folloving characteristics: Acid number of 0.3; Saponification number of 47.4; Hydroxy value of 42.5; pH (5X) of 4.6; Color APHA = 40; m.p. = 36.0eC; Freezing point - 32.4’C; H20 content (X, KF) - 0.03. d3 Polyoxyethylene-sorbitan-fatty acid esters or polysorbates e.g. of the type knovn and commercially available e.g. under the Trade Marks Tveen and Armotan (c.f. Fiedler, 2, p.p. 745-746 and 972-975) including the products Tveen - 18 20 [polyoxyethylene(20)sorbitanmonolaurate], I polyoxyethylene(20)sorbltanmonopalmitate], [polyoxyethylene(20)sorbitanmonostearate], (polyoxyethylene(20)sorbitantristearatej, g 80 [polyoxyethylene(20)sorbitanmonooleate], lpolyoxyethylene(20)sorbitantrioleatel, [polyoxyethylene(4)sorbitanmonolaurate], [polyoxyethylene(4)sorbitanmonostearate], and 81 [polyoxyethylene(5)sorbitanmonooleate]. d4 Polyoxyethylene fatty acid esters, for example polyoxyethylene stearic acid esters of the type knovn and commercially available under the Trade Mark Myrj (c.f. Fiedler, 2, p. 636) as veil as polyoxyethylene fatty acid esters knovn and commercially available under the Trade Mark Ig Cetiol HE. (c.f. Fiedler, 1, p. 228), and polyoxyethylene fatty alcohol ethers, for example polyoxyethylene stearylether, oleyl ether, or cetyl ether, e.g. of the type knovn under the Trade Mark Brij (c.f. Fiedler, 1, 184-186), e.g. Brij 78 and 96, and Cetomacrogel 1000 (c.f. Fiedler, 1, 229). d5 Polyoxyethylene-polyoxypropylene co-polymers e.g. of the type knovn and commercially available under the Trade Marks Pluronic and Emkalyx (c.f. Fiedler, 2, p.p. 720-722. 2g d6 Sorbitan fatty acid esters e.g. of the type knovn and commercially available under the Trade Mark Span, for example including sorbitanmonolauryl, -monopalmityl, -monostearyl, -tristearyl, -monooleyl and -trioleyl esters - c.f. Fiedler, 2, p.p. 850-851. 3θ d7 Partial glycerides, e.g. products comprising mono- and/or di-glyceride components, for example mixed mono-/di-/tri-glyceride/free glycerol based surfactants as veil as, e.g., glycerol monostearates and glycerol monooleates. Examples of such products include those knovn and commercially available under the Trade Marks Imvitor (c.f. Fiedler, 1, 491) for example Imvitor 191 and Imvitor 780< and Softigen (c.f.

Fiedler, 2, 833) for example Softigen 701. - 19 de Ionic surfactants or emulsifying agents such as sodium lauryl sulfate and sodium cetostearyl sulfate.

Components (d) when present are suitably present in an amount of up to 5 maximally 30%, more preferably maximally 15X, based on the total weight of the composition. Most suitably components (d) are present in an amount of from about 0.5 to about 10X, e.g. about 1.0X, based on the total weight of the composition.

IQ Such compositions will suitably also comprise: e) A thickening agent.

Suitable components (e) include for example: ex Polymethylacrylate resins, e.g. of the type known and commercially available under the Trade Mark Eudispert (c.f. Fiedler, 1, 371-372). e2 Cellulose derivatives including e.g. ethyl-, propyl-, methyl-, hydroxy20 propylmethyl- and carboxymethyl-celluloses. e3 Polyvinyl resins, e.g. including polyvinylalcohols and polyvinylpyrrolidones, as veil as other polymeric materials including gelatin, alginates, pectins, gum traganth, gum arabicum, gum xanthan, and polyacrylic acids such as known and commercially available under the Trade Mark Carbopol (c.f. Fiedler, 1, 206-207). e* Materials such as silica gel, bentonite, and magnesium-aluminium silicate.

Components (e) when present are suitably present in an amount of up to 20X, more preferably up to 10X, based on the total weight of the composition. Most suitably components (e) are present in an amount of from about 0.5 to about 15X, e.g. from about 1.0 to •ir about 3.OX based on the total weight of the composition.

In addition such compositions also suitably comprise an anti-microbial agent such as methylparaben, propylparaben, benzalconium chloride or benzyl alcohol, e.g. in an amount of from 0.05 to 0.52 or, in the case of benzyl alcohol, up to 2.02, e.g. ca. 0.12, or, in the case of benzyl alcohol, ca. 1.02 by weight based on the total weight of the composition.

Alternative carrier media include hydrophobic materials, e.g. which meet the solubility criteria with respect to the active ingredient hereinbefore described. Further preferred compositions in accordance with the invention, e.g. for topical application, accordingly comprise C2 a) Ciclosporin in non-solvate orthorhombic crystal form, i.e.

Cy-A/X-III, e.g. as defined under anyone of A3 to A7 above; and f) A hydrophobic carrier, e.g. as set forth above.

The amount of component (a) present will suitably be as described above in relation to compositions as defined under C1.

Suitable components (f) include, for example, petroleum derivatives, including mineral oils, fats and jellys, as for example petrolatum (mineral fat or petroleum jelly) liquid petrolatum (white mineral oil or liquid paraffin), or any of the products known and commercially available under the Trade Mark Vaseline [c.f. Fiedler loc. cit. 2, 986], as well as non-ionic lanoline based derivatives such as available under the Trade Mark Amerchol, e.g. Amerchol CAB [Fiedler, 1, 119-120].

Further suitable components (f) include, for example natural or saturated vegetable oils and fats, e.g. fractionated coconut oils as known and commercially available under the Trade Mark Miglyol, e.g. Miglyol 812 (c.f. Fiedler, 2, 616) and peanut and saturated pea-nut oils, as well as higher fatty or branched chain alcohols and fatty esters, such as stearic alcohol, cetyl palmitate and 2 -octyldodecanol which is known and commercially available under the Trade Mark Eutanol G (c.f. Fiedler, 1, 375).

Such compositions may also comprise one or more of any of components (d) and (e) hereinbefore described, as veil as one or more anti-microbial agents, e.g. as hereinbefore described.

In particular such compositions may additionally comprise one or more components (d), suitably in an amount of up to at most 30X, more preferably at most 20%, suitably in an amount of from about 1 to about 15% by veight based on the total veight of the composition.

Hydrophobic based compositions in accordance vith C2 and comprising an emulsifier component (d), may also be prepared in the form of creams or emulsions by the further addition of vater.

The obtained creams or emulsions may be either of the oil-in-vater or vater-in-oil type. The amount of vater present in such compositions may thus vary from e.g. 5 to 80% based on the total veight of the composition,, suitably from 20 to 70%.

Amounts of components (e)/antimicrobial agent, vhen incorporated, vill be of the same or similar order to those set forth in relation to compositions of the invention in accordance vith C1 above.

The amount of any topical form applied vill, of course, vary depending on the concentration of the composition, the condition to be treated and^the effect desired, as veil , in the case of dermal application, the area to be covered. In the case of compositions for dermal application, e.g. in the treatment of psoriasis or atopic dermatosis, amounts applied vill generally be sufficient to provide a Cidosporin dosaging of the order of from about 0.1 to about 5mg/cm2, e.g. ca. 0.5mg/cm2, vith administration e.g. 2x daily to the desired site. In the case of ophthalmic preparations, administration may suitably be performed employing preparations comprising about 0.1 to about 10%, e.g. 1.0 or 2.OX Ciclosporin by weight, applied in drop form to each eye 1 to 3x daily.

INJECTIBLE FORMS, e.g. for sub-cutaneous, intra-muscular or other parenteral injection, Including, in particular, intra-articular injection, e.g. in the treatment of arthritic disease such as rheumatoid arthritis, or for intra-lesional Injection, e.g. in the treatment of psoriasis.

Dispersions, suspensions, emulsions and like compositions suitable for injection can be prepared in accordance vith any of the techniques knovn and generally employed in the art, for example by dispersing, suspending or otherwise distributing Ciclosporin in fine particulate orthorhombic crystal form in an appropriate liquid carrier medium vith the addition of appropriate emulsifiers, sufactants, stabilizing or flocculating agents etc... Preferably the Ciclosporin component in such compositions vill have an average particle size of <100uM, more preferably <50μΜ, most preferably <20μΜ. Suitably the average particle size vill be of the order of from 0.1 or 0.5 up to the indicated 20, 50 or ΙΟΟμΜ maxima.

Vhere injectible forms are intended e.g. for sub-cutaneous or intra-muscular injection, for example to achieve a depot affect, use of larger particle size vill be possible. Vhere such forms are intended for intra-lesional application or, in particular, intra-articular injection, use of finer particule preparations vill be indicated. For such uses a suitable average particle size is e.g. of the order of from about 0.1 or about 0.5 to about 20μΜ, e.g. ca. 5 to 15μΜ.

To achieve stability of particles in injectible forms, e.g. to avoid increase in size of larger crystalline particles at the expense of smaller particles, the particles vill preferably be of as uniform a size as possible. Suitably variation betveen maximum and minimum particle size vill not exceed about 50μΜ, more preferably about 20μΜ.

Although the desired particle size may be achieved by use of appropriate comminution procedures, e.g. micronisation, to minimize particle size variation, to avoid the presence of fragmented particles, to avoid modification of the crystal surface, e.g. to retain wettability characteristics, and to more easily meet required sterility requirements, it will generally be preferred to employ non-comminuted microcrystals, i.e. crystals grown to the desired size requirements as hereinbefore described.

Injectible compositions in accordance with the invention thus appropriately comprise: a) Ciclosporin in fine particulate non-solvate orthorhombic crystal form, i.e. CY-A/X-III, e.g. as defined under any one of A3 to A7, in particular A6 and A7, above, distributed or distributable within, b) an injectible carrier medium.

Suitable carrier media include, in particular, pharmaceutical grade water.

In addition compositions as defined under C3 vill suitably comprise g) a sufactant as dispersant or emulsifying agent and/or a flocculating agent, to assist or maintain distribution of the particles throughout the carrier phase.

Suitable surfactant components (g) include polyoxyethylene-sorbitan fatty acid esters or polysorbates, e.g. as set forth under (d3) above, e.g. Tween 80 or Polysorbate 80 (c.f. Fiedler, 2, p.p. 746). Suitable flocculating agents (g) include ethylene diamine tetra-acetate and salts thereof, for example Calcium EDTA and Sodium EDTA, as veil as other equivalent chelating agents, and peptisating agents such as succinic or citric acid.

In order to maintain particle size distribution, a stabilizing agent is.also suitably incorporated. Suitable stabilizing agents - 24 include e.g. gelatins or modified gelatins such as the plasma expander knovn and commercially available under the Trade Mark Gelafundin and Haemaccel or cold-vater soluble gelatins of highly purified collagen hydrolysates.

Compositions in accordance vith C3 above may also include one or more anti-microbial agents, e.g. as hereinbefore described, as veil as peptisating agents such as succinic or citric acid.

The ratio of components (a):(b) in compositions as defined under C3 is suitably of the order of 1:10 to 1:1,000, preferably of the order of from 1:50 to 1:200 e.g. about 1.100 p.p.v.

In order not to unduly disturb particle stability, surfactants present as component (g) vill preferably be present in only minor concentrations, e.g. of the oder of from 0.1 to 2.OX, suitably ca. IX by veight based on the veight of component (a). Where a flocculating agent is present the ratio of flocculating agent to component (a) is suitably of the order of from 1.3 to 1:8 p.p.v. e.g. ca. 1:5 p.p.v. When a stabilizing agent is present, the ratio of component (a)stabilizing agent is suitably of the order of from 1:5 to 1:30, more preferably 1:10 to 1:30, e.g. ca. 1:20 p.p.v. When a peptisating agent is present, the ratio of peptising agent to component (a) is suitably of the order of from 1:1 to 1:10, e.g. ca. 1:15 p.p.v.

The amount of Ciclosporin present in unit dosage forms for injection in accordance vith the invention vill of course vary depending on e.g. the condition to be treated, the site of injection and the effect desired, e.g. vhether depot effect or relatively speedy release into the surrounding tissues is contemplated. Suitably individual unit dosage forms vill comprise ca. 10 to 500 mg, especially from ca. 20 to lOOmg Ciclosporin/dosage. - 25 10 As will be appreciated, unit dosage forms in accordance vith the invention also include more complex systems, e.g. double chambered syringes or syringe barrels in vhich the particulate Ciclosporin together vith other appropriate components, e.g. stabilizer or anti-microbial agent, is retained in a first chamber and the carrier medium is retained in a second chamber, the components of the tvo chambers being brought into admixture, e.g. folloving actuation of the syringe plunger. Such double chamber syringe systems are veil known in the art and unit dosage forms comprising them are to be understood as being within the purview of the present invention. - 26 Ingredients referred to herein or defined as components of pharmaceutical compositions in accordance with the invention are to be understood as being suitable for pharmaceutical application as required, e.g. as being pharmaceutically acceptable or, in the case of ingredients of compositions for topical administration, as being topically, e.g. dermally or occulary, applicable, acceptable or tolerable.

Further galenic forms for administration by equivalent or alternative routes to those above described, will be apparent to the skilled worker.

The following examples are illustrative of the present invention in each of its various aspects. - 27 EXAMPLE 1 PREPARATION OF Cy-A/X-III a) 100 g of Cidosporin in amorphous form are dissolved vith stirring in 400g polyethylene glycol 300 at 50°C. CY-A/X-III development commences shortly after completion of dissolution. The solution is alloved to stand for 24-48 hrs. at ca. 35-40 °C to complete crystallisation and then diluted vith 1000 ml vater vith vigorous stirring. The obtained precipitate is separated by filtration, vashed vith H30 to remove remaining polyethylene glycol and dried to yield CY-A/X-III in substantially pure form: m.p. = 195 °C, particle size = ca. 100-200 uM.

CY-A/X-III forms may be prepared analogously to the above procedure but employing the folloving materials as solvent in place of polyethylene glycol 300.

SOLVENT M.P. FOR OBTAINED CY-A/X-III (°C) b) Polyethyleneglycol 300/ Cremophore RH 40 (1:1 by volume) c) Cremophore EL 190 d) Solutol* HS 15 188 e) Gelucire* 44/14 180 f) Myrj 52 184 g) Tveen 80 * 186 h) Cremophor RH40 190 (R = Registered Trade Mark) In each case the obtained average particle size is of the order of from about 50 to about 200 uM. Determined lattice structure is as hereinbefore defined. X-ray diffraction data are as hereinafter described vith reference to Table III and Fig. IV. - 28 EXAMPLE 2 PREPARATION OF CY-A/X-III 20g Ciclosporin as CY-A/X-I is dissolved in 60ml ethanol and condensed to l/6th of the initial volume by distillation under reduced pressure at >60°C. 160ml polyethylene glycol (MV 300) + 20ml H20 are then added slowly at 50-85°C. vith stirring at 300 r.p.m.. An ultrasonic vibrator/mixer is installed operating at 20,000 c.p.c. Nucleation is initiated at 65-75eC. over 3 to 5 minutes. The obtained dispersion is then cooled linearly over 10-20 minutes to 40°C. with continued stirring at 700 r.p.m. and ultrasonic vibration. The obtained thick pasty mixture is filtered, rinsed with polyethyleneglycol (MV 3OO)/H2O (3:1 p.p.vol), washed with warm water (30eC.) and dried at 50eC. under high vacuum, to yield CY-A/X-III in pure form : m.p. 192°C. Particle size = 3-15uM. Determined lattice structure as hereinbefore defined. X-ray diffraction data as described hereinafter vith reference to Table III and Fig. IV.

Repeat experiments indicate that crystal size is influence by nucleation temperature and rate of crystal grovth rather than duration of processing. Thus by initiating nucleation at ca. 75°C. and continuing crystallisation at the same temperature, an average crystal particle size of ca. 50uM is obtained.

EXAMPLE 3 PREPARATION OF GALENIC FORMS FOR TOPICAL APPLICATION 3-1 GEL PREPARATION I Cy-A/X-III obtained in accordance with the procedures of Example 1 is micronised employing an air-jet mill. Micronised product having a maximum particle size of 60-70uM and an average particle size of from 2-10μΜ is combined and mixed in conventional manner with the ingredients shown in the table below to provide an aqueous gel comprising 10 2 by weight Ciclosporin (as Cy-A/X-III) and suitable for topical application, e.g. for the treatment of psoriasis.

INGREDIENT 2 BY WEIGHT a) CICLOSPORIN (micronised Cy-A/X-III) 10.0 b) Polyacrylic acid (Carbopol 934P) 0.5 c) NaOH-solution (52) 3.0 d) Methylparaben 0.07 e) Propylparaben 0.03 f) H20 (pharmaceutical grade) to 100.0 2 3.2 GEL PREPARATION II INGREDIENT X BY VEIGHT Composition A Composition B a) CICLOSPORIN (micronised Cy-A/X-III) b) Polyethylenglycol 300 c) Labrafil M 2130 CS d) Carbopol© 934 P e) Sodium hydroxide 5% f) Methylparaben g) Propylparaben h) H20 (p.g.) .0 .0 1.0 1.0 6.0 0.07 0.03 51.9 1.0 .0 1.0 1.0 6.0 0.07 0.03 60.9 (R « Registered Trade Mark) 100.0 X 100.0 X a) is prepared as for 3.1 above, c), f) and g) are dissolved in b) vith varming and the obtained mix cooled to toom temperature. This solution is on then dispersed in vater h). d) is added and the vhole homogenized to effect dispersion, a) is added and the mix again homogenized, e) is added to neutralise, vith stirring, and the resulting gel filled into collapsible tubes for topical application, e.g. in the treatment of psoriasis. 3.3 OINTMENT PRAPARATION ' INGREDIENT a) CICLOSPORIN (micronised Cy-A/Xb) Amerchol© CAB c) Liquid paraffin d) Vhite petrolatum 35 (© Registered Trade Mark) X BY VEIGHT Composition A Composition 1.0 10.0 2.0 2.0 42.0 37.5 55.0 50.5 100.0 X 100.0 X -31 a) is prepared as for 4.1 above, b), c) and d) are melted together and stirred and a) added at ca. 30’C, the particles being distributed employing an homogenizer. The obtained ointment is cooled and filled into collapsible tubes for topical application, e.g. in the treatment of psoriasis. 3.4 CREAM PREPARATION INGREDIENT X BI VEIGHT a) CICLOSPORIN (micronised Cy-A/X-III) b) Polysorbate 60 c) Cetylstearyl alcohol d) Glycerol 85X 1.0 5.0 10.0 10.0 e) Vhite petrolatum 24.0 f) Methylparaben 0.07 g) Propylparaben 0.03 h) H20 (p.g.) 49.0 100.0 X a) is prepared as for 3.1 above, combined vith components b) to h) analogously to 3.3 above and filled into collapsible tubes for topical application, e.g. in the treatment of psoriasis. 3.5 OINTMENT PREPARATION A suspension comprising 20 X by veight of Cy-A/X-III (obtained in accordance vith the procedures of example 1) in polyethyleneglycol 300 is vet-milled until the average Cy-A/X-III particle size is of the order of from 2 to 10 μΜ. lOg of the obtained milled product is mixed in conventional manner vith 50g polyethyleneglycol 300 and 40g melted polyethyleneglycol 1500 to yield a flowable paste comprising 10 X by veight ciclosporin (as Cy-A/X-III) and suitable for topical application, e.g. for the treatment of psoriasis. -323.6 GEL PREPARATION INGREDIENT X BY WEIGHT a) CICLOSPORIN (micronised CY-A/X-III) 1.0 b) Labrafil M 1944 CS 1.0 c) Polyethylene glycol 300 30.0 d) Benzyl alcohol 1.0 e) Hydroxypropylmethylcellulose (Methocel E4M) 2.5 f) H20 (p.g.) 64.5 100.0 X a) is prepared as for 3.1 above, b), c) and d) are mixed, and f) added and mixed, a) is then suspended in the resulting mixture by homogenisation, e) is then added slowly vith stirring until gel formation is complete. The gel is then filled into collapsible tubes for topical application, e.g. in the treatment of psoriasis. 3.7 GEL PREPARATION INGREDIENT X BY VEIGHT a) CICLOSPORIN (micronised CY-A/X-III) 5.0 b) Polyethyleneglycol 300 30.0 c) Glyeerylmonostearate 5.0 d) Methylparaben 0.07 e) Propylparaben 0.03 f) Carbopol 934 P 1.0 g) Sodium hydroxide 5X 6.0 h) H20 (p.g.) 52.9 100.0 X 33a) is prepared as for 3.1 above, c), d) and e) are dissolved in b) with warming. The warm mixture is dispersed in h) by homogenisation and cooled to room temperature, a) and f) are then added and the whole homogenised, g) is then added and gel formation completed. The obtained gel is filled into collapsible tubes for topical application, e.g. in the treatment of psoriasis. 3.8 CREAM PREPARATION (WATER IN OIL) INGREDIENT X BY WEIGHT a) CICLOSPORIN (micronised CY-A/X-III) 2.0 b) Amerchol CAB 5.0 c) Miglyol 812 10.0 d) White petrolatum 53.0 e) H20 (p.g.) 30.0 100.0 X a) is prepared as for 3.1 above and dispersed with b) in e) while warming and homogenising, c) and d) are molten together, mixed and the mixture added to the previously obtained dispersion with intensive homogenisation, and the whole cooled to provide a fatty white cream. This is filled into collabsible tubes for topical application, e.g. in the treatment of psoriasis. 3.9 GEL/CREAM PREPARATIONS 30 Examples 3.1 to 3.8 above are repeated but employing the crystalline product of example 2 directly as CICLOSPORIN component. - 34 EXAMPLE 4 4.1 INJECTIBLE FORM, E.G. FOR PARENTERAL APPLICATION A Cy-A/X-III suspension is prepared under sterile conditions, by conventional techniques and employing the folloving ingredients lQ INGREDIENT VT (mg) a) CICLOSPORIN (CY-A/X-III) 20.0 b) Polysorbate 80 4.0 c) Sodium carboxymethyl cellulose 5.0 d) NaCl 9.0 e) Benzyl alcohol 9.0 f) H20 (injection grade) to an end volume of 1.0 ml The active ingredient release characteristics of the obtained formulation are dependent upon the average particle size of component a). Thus vhere activity over longer periods of time is required, Cy-A/X-III product of large average particle size (e.g. 100-200 μΜ as obtainable directly via the procedures of example 1) vill be chosen. Vhere activity over relatively shorter periods of time is required, Cy-A/X-III product of relatively 25 smaller average particle size (e.g. obtained by micronisation or milling of Example 1 product, e.g. to a particle size of from 0.5-10 μΜ) vill be chosen. The obtained suspension is filled into injection ampoules for parenteral administration or for intra-lesional injection in the treatment of psoriasis, for example to provide a depot effect.

In an especially preferred embodiment the example is performed employing CY-A/X-III prepared in accordance vith example 2 above, having a particle size of from 5-15μΜ, and prepared under sterile conditions. 354.2 INJECTIBLE FORMS, E.G. FOR INTRA-ARTICULAR INJECTION Injectible forms suitable for intra-articular administration are prepared 5 employing the folloving ingredients: INGREDIENT Composition A VT (mg) t5 a) CICLOSPORIN (CY-A/X-III) 10.0 b) Sodium carboxymethyl cellulose 10.0 c) Na EDTA 2.0 d) H20 (injection grade) to an end volume of 1.0 ml Composition B a) CICLOSPORIN (CY-A/X-III) 10.0 b) Polysorbate 80 0.1 c) Citric acid 2.0 d) H20 (injection grade) to an end volume of 1.0 ml Formulation is carried out in accordance vith standard techniques under sterile conditions. Component (a) comprises non-comminuted micro-crystals, produced under sterile conditions in accordance vith the procedures of example 2 and having a particle size of from 3-15yM. The obtained compositions are filled, again under sterile conditions, into ampoules for injection, e.g. intra-articular injection for the treatment of rheumatoid arthritis.

The advantageous properties of compositions in accordance vith the invention may be demonstrated in animal test models as veil as in clinic.

Thus particular utility of compositions in accordance vith the invention intended for dermal application, e.g. in the treatment of psoriasis or dermatosis, may be demonstrated in the folloving test model: - 36 ALLERGIC CONTACT DERMATITIS IN THE GUINEA PIG (DTH-TEST) Guinea pigs (Hartley, σ’, 400-500 g) are sensitized by application of 50 ul of 2 X dinitrofluorobenzene (DNFB) in acetone/olive oil (1:1) to the inner side of the right ear. 7 days later, the animals are re-treated vith 20 ul, 0.5 X DNFB in acetone/olive oil (4:1) applied to marked areas on the shaven, left and right flank. This second challenge exposure induces an allergic inflammation, leading to reddening and cellular infiltration (thickening) of the skin. Test composition in an amount of from 200-250 mg is applied vith a spatula to the DNFB treated area of the right flank. The left flank is similarly treated vith placebo as control. Application of test composition/placebo is effected 5x at intervals of 20 mins, 8hrs., 24 hrs., 32 hrs., and 48 hrs., after the challenge. Skin thickness at the site of application is determined before each application, and again 8 hrs. after the last application, by raising the skin into a fold and measuring the thickness of this. Degree of reddening or inflammation is also estimated visually on a scale of from 0 to 4. Efficacy of test preparation in preventing inflammatory response is determined by comparison vith results recorded for placebo treated flanks.

Results obtained in the above test model employing compositions in accordance vith examples 31 3.5 above but comprising. 0.1 X by veight Ciclosporin as CY-A/X-III, in comparison vith placebo and vith topically applied 0.1 X Ciclosporin in convention oral solution form are as follovs. (A 0.1X CY-A/X-III concentration is employed in this test model in viev of its extreme sensitivity. Test compositions are prepared entirely analogously to examples 3.1 and 3.5 but by reduction of the Ciclosporin component to O.lg and compensation by addition of 9.9g vater in the case of example 3.1 and polyethyleneglycol components in the case of example 3.5.] In each figure, measured skin thickness is plotted against time. In Fig. I results employing 0.1X (by veight) Ciclosporin in conventional applied in the form of the conventional, commercial oral, drink solution form are compared vith placebo. Slight but insignificant reduction in reaction as compared vith placebo is evidenced for the oral solution. In Figs. II and 37III, results employing 0.1% (by weight) Ciclosporin containing compositions in accordance with examples 3.1 and 3.5 respectively are compared with placebo. In both instances, substantial reduction in skin thickening as compared with placebo are achieved following first application and continuing through treatment as compared with placebo are achieved following first application and continuing through treatment until completion of the experiment.

IO Utility of compositions in accordance with the present invention may also be demonstrated in clinical trials e.g. performed as follows: CLINICAL TRIAL: TREATMENT OF PSORIASIS BY TOPICAL (DERMAL) APPLICATION The trial is performed in randomized, placebo controlled, double-blind design. Each patient receives active composition and placebo (vehicle alone) on two symmetrical lesions simultaneously. Allocation of active and placebo compositions is randomised to the left and right lesions.

Selected subjects are taken in groups of from 12 to 24 patients, aged 18-70 years including males and females who are postmenopausal, surgically sterile or using oral contraceptives with a negative pregnancy test.

Inclusion criteria include: a) written informed consent and b) clinically defined, stable, chronic plaque psoriasis. At least two moderate to severe bilateral symmetrical lesions, each having an area of 4 to 25cm:! and a severity score >5 on a scale ranging from 0 to 9. This score is a composite of individual scores of 0 (least severe) to 3 (most severe) for each of three parameters: erythema, scaling and skin thickness. Lesions on both sides must be comparable within the same patient (i.e. not differing by more than 20% in severity score or area).

The following exclusion criteria are applied: a) Any medical condition which, in the opinion of the investigator, would compromise the study. - 38 b) Systemic psoriasis therapy.

(PUVA, methotrexate, steroids, retinoids, ciclosporin) vithin the last 2 months. c) Specific topical treatment - except indifferent substances (e.g. vhite vaselin) or formulations containing salicylic acid (not more than 10X) on lesions selected for study vithin the last 2 weeks before entering the study. These emollients may however be used on lesions not selected for the study. d) Patients whose psoriasis appears to be spontaneously improving, flaring without treatment, or rebounding from cessation of therapy. e) Known hypersensitivity to any of the ingredients of the test medication. f) Uncooperative patients who are unlikely to follov medical instructions exactly, patients not willing or not able to attend regular visits. g) Treatment vith an investigational drug vithin one month before study enrollment. h) Severe concomitant disease, especially renal dysfunction vith serum creatinine above 130μΜο1/1. i) Hypertension (diastolic BP> 95mm Hg after 5 minutes in sitting position). Patients vith drug controlled hypertension are included into the study. j) Malignancy or history of malignancy, including skin cancer. k) Acute bacterial, viral, or fungal associated skin lesions. l) Pregnancy, lactation. m) Patients vith a history of alcoholism, drug abuse, psychosis, or emotional or intellectual problems. n) Patients vho need continuous concomitant therapy vith other immunosuppressants, chemotherapeutic agents, compounds vith a nephrotoxic potential (e.g. aminoglycosides) and drugs vhich are knovn to interact vith the pharmacokinetics of Ciclosporin (ketoconazole, erythromycin, phenytoin, phenobarbitone, rifampicin, INH, carbamazepine, sodium valproate). o) Patients vith abnormal baseline laboratory values > 15X outside normal limits for the testing laboratory. p) Patients vith clinically significant findings in physical examination.

Trial compositions employed are in accordance vith any one of examples 3.1 to 3.9 hereinbefore described, e.g.: the IX composition B of example 4.2 or the 10X composition B of example 3.3.

Each patient receives a pair of 20g coded tubes bearing a tvo-panel label. The label is blue for the substance vhich has to be applied on the right side and red for the left side. Further, the side to be treated, left or right, the study code, the investigator name, the patient number, and the date of starting treatment, are vritten on each panel of the label. The tear-off panel of the label is affixed in the case report form.

Test medication is applied 2x daily, morning and evening, the amount applied depending on lesion size, to give a daily Cidosporin dosaging of from 0.2 to 2.0mg Ciclosporin/cm2, e.g. in one test series ca. 0.5mg Ciclosporin/cm2. Placebo is applied in equivalent amount. Neither trial composition nor placebo is applied to a body surface >25cm2 and no more than lg Cidosporin is applied per veek. At each clinic visit the medication/placebo tubes are veighed and the topically applied dally dose determined. Subjects may bathe shortly before application of medication.

Disposable gloves are put on and changed to apply each trial composition and placebo to avoid cross contamination. At least three hours are alloved to elapse betveen administration and vashing of the treated areas. The first application is supervised and a patient instruction sheet is provided. Duration of treatment is 4 veeks. If complete clearance of lesions occurs earlier, treatment is continued until veek 4. Patients are investigated before trial commencement (veek -1), at the beginning of the trial (veek 0) and after 1, 2, 3 and 4 veeks.

* Dermatological assessment (local psoriasis severity index or LPSI) is assessed by the same investigator, not involved in medication, using the folloving 0-3 numeric scales. - 40 Erythema Ο = None = Minimal: faintly detectable erythema; very light pink « Moderate dull red distinguishable = Strong: deep/bright red Scaling = none = Minimal: poorly defined scales Moderate: veil-defined scales starting to pile up 3« Severe: veil defined scales; piled up to compact Skin Thickness = none = Minimal: barely perceptible elevation - Moderate: elevation clearly perceptible = Severe: high elevation The total LPSI range value of erythema plus scaling plus thickness - 0 to 9 is evaluated. To enter into the study, at least 2 signs must be moderate to severe for both selected lesions (LPSI >5).

At entry and at each clinic visit, the area of the treated lesions is evaluated (Entry size: 4 to 25cm2 for each plaque). Area refers to presence of one or more of the three criteria, erythema, scaling, thickening. Pigmentation vithin the lesion area is not considered in area estimation The plaques are registered by photographs before (week -1) and after the treatment (veek 4). Three pictures are taken: one shoving both lesions together and one for each lesion.

Pruritus is evaluated from patient interviev using the folloving scale: = none = minimal: occasionally has urge to scratch = moderate: has urge to scratch frequently - 41 3 = severe: has urge to scratch very frequently, may disturb sleep.

At end of the study, a global assessment of response to therapy is made in accordance to the folloving criteria: - cleared = 100% resolution - marked improvement at least 66% reduction of LPSI, and/or at least 66% reduction of plaque area - moderate improvement: between 65% and 33% reduction of LPSI, and/or between 65% and 33% reduction of plaque area - poor: less than 32% reduction of LPSI and/or less than 32% reduction of plaque area - worse: exacerbation of disease Therapeutic success is defined as any lesion which has cleared or which exhibits marked improvement.

Physician and patient also indicate their preference for left or right side medication in accordance with the folloving criteria: · left side definitely better . left side slightly better . no difference detectable . right side slightly better . right side definitely better To test efficacy at the end of treatment, subject LPSI scores and lesional areas are compared by means of the VILCOXON-Sign Rank test for paired treatments betveen Ciclosporin-treated sides and placebo treated sides with respect to change in score from baseline to after 4 weeks of treatment.

All further comparisons (at other time points and for the remaining parameters of efficacy), including the p-values attached to them, are taken as descriptive of progress in trial. in the above trial, subjects exhibit marked improvement in psoriatic lesion, e.g. of LPSI scores for erythema, scaling and skin thickness, on 42sides to vhich test composition comprising Ciclosporin is applied as compared vith side receiving placebo. Subjective improvement is also recorded, together vith improvement in recorded values for pruritis on Ciclosporin treated as compared vith placebo treated sides.

Positive results are also obtained in trials of parallel design for subjects exhibiting contact dermatitis as diagnosed via 3 or more of the folloving criteria: - Pruritus - Typical morphology and distribution: Flexural lichenification or linearity in adults Facial and extensor involvement in infants and children - Chronic or chronically-relapsing dermatitis - Personal or family history of atopy (asthma, allergic rhinitis, atopic dermatitis); plus 3 or more minor features including: Xerosis, ichthyosis/palmar hyperlinearity/keratosis pilaris, immediate (type I) skin test reactivity, elevated serum IgE, early age of onset(< 5 years), tendency tovard cutaneous infections (esp. Staph, aureus and Herpes simplex)/ impaired cell-mediated immunity, tendency tovard non-specific hand or foot dermatitis, nipple eczema, cheilitis, recurrent conjunctivitis, Dennie-Morgan infraorbital fold, keratoconus, anterior subcapsular cataracts, orbital darkening, facial pallor/facial erythema, pityriasis alba, anterior neck folds, itch vhen sveating, intolerance to vool and lipid solvents, perifollicular accentuation, food intolerance, course influenced by environmental/emotional factors and vhite dermographism/delayed blanch. The trial period is three veeks, vith evaluation on a scale of 0 to 3 for the folloving parameters: pruritis, erythema, exudation, excoriation and lichenification.

Topical formulations as hereinbefore described in examples 3.1 to 3.9, containing 1-10X Ciclosporin, for example the 5.OX composition in accordance vith example 3.7 are also found to be effective in the treatment - 43 of allergic contact dermatitis as veil as of alopecia.

In accordance vith the foregoing CY-A/X-III is indicated for use in: D A method of effecting immunosuppressive or anti-inflammatory therapy in a subject in need thereof, vhich method comprises administering to said subject an effective amount of Ciclosporin in non-solvate orthorhombic crystal form, i.e. as CY-A/X-III, e.g. as defined under any one of A3 to A7 above.

Purposes for vhich the above method is applicable include treatment of any disease or condition as hereinbefore described in relation to use of Ciclosporin, e.g. prevention of transplant rejection, treatment of auto-immune or rheumatic disease, treatment of any other disease or condition involving undesirably high response of the immune system, treatment of parasitic infection, treatment of chemotherapeutic resistance, e.g. reversal of tumor resistance to anti-neoplastic chemotherapy, as veil as the treatment of alopecia. The above methods are in particular applicable to the treatment of diseases or conditions of the skin or eye, e.g. psoriasis, atopic or contact dermatitis, corneal transplant, conjunctivitis and uveitis, by topical administration, as veil as to the treatment of rheumatoid arthritis or other disease or condition of the joint, by intra-articular administration.

The present invention accordingly provides: E. Ciclosporin in non-solvate orthorhombic crystal form, i.e. as CY-A/X-III, e.g. as defined under any one of A3 to A7 above, for use as a pharmaceutical, e.g. for use in the treatment of any disease or condition as hereinabove set forth in relation to D above, for example, for use as an immunosuppressive or anti-inflammatory agent; as veil as 5 F. Ciclosporin in non-solvate orthorhombic crystal form, i.e. as CY-A/X-III, e.g. as defined under any one of A3 to A7 above, for use in the preparation of a pharmaceutical compostiion for use as defined under E above.

To assist identification of Cyclosporin in non-solvate orthorhombic crystal form, i.e. CY-A/X-III, X-ray povder diffraction data for CY-A/X-III and for the tvo forms CY-A/X-I and CY-A/X-II also referred to herein, are provided in the folloving Tables I to III. Fig. IV attached provides a representation of the X-ray povder diffraction diagram from vhich the data in Tables I to III vere obtained. The diffraction diagram vas obtained employing a Guinier-DeVolff II camera, using CuKa-radiation, X - 1.542 A. \ - 45 TABLE I X-RAY DIAGRAM FOR CY-A/X-I Line No. d (A) Intensity 1. 13.0 VS 2. 11.4 VS 3. 10.3 VS 4. 9.7 VS 5. 9.4 VS 6. 8.25 VS 7. 7.1 S 8. 6.1 M 9. 5.85 S 10. 5.6 S 11. 5.25 VS 12. 4.81 S 13. 4.58 S 14. 4.25 M 15. 4.0 M 16. 3.67 M 17. 3.45 * M - 46 TABLE II X-RAY DIAGRAM FOR CY-A/X-II Line No. d (A) Intensity 1. 17.9 S 2. 11.4 VS 3. 11.0 S 4. 10.6 M 5. 10.2 S 6. 8.9 M 7. 8.7 M 8. 8.1 M 9. 7.2 VS 10. 6.3 M 11. 5.95 M 12. 5.7 M 13. 5.35 M 14. 5.1 S 15. 4.85 S 16. 4.8 M 17. 4.55 M 18. 4.4 M - 47 TABLE III X-RAY DIAGRAM FOR CY-A/X-III Line No. d (A) Intensity 1. 12.0 M 2. 10.4 VS 3. 9.6 s 4. 8.7 s 5. 7.9 M 6. 7.7 s 7. 6.7 M 8. 6.0 M 9. 5.83 s 10. 5.3 M 11. 5.2 M 12. 4.92 S 13. 4.88 S 14. 4.58 M 15. 4.48 M 16. 4.0 M 17. 3.59 M 18. 3.38 M VS = very strong S & strong M s medium

Claims (31)

1. Ciclosporin in non-solvate orthorhombic crystal form. 5

2. Ciclosporin in non-solvate orthorhombic crystal form having a crystal lattice « a = 12.7 A, b = 15.7 A, c = 36.3 k, volume per asymmetric unit = 1804 A.

3. Ciclosporin in non-solvate orthorhombic crystal form exhibiting the folloving X-ray povder diffraction characteristics: Line No. d (A) Intensity 1. 12.0 M 2. 10.4 VS 3. 9.6 s 4. 8.7 s 5. 7.9 M 6. 7.7 S 7. 6.7 M 8. 6.0 M 9. 5.83 S 10. 5.3 M 11. 5.2 M 12. 4.92 S 13. 4.88 S 14. 4.58 M 15. 4.48 M 16. 4.0 M 17. 3.59 M 18. 3.38 M - 49 VS = very strong S = strong M = medium

4. Ciclosporin according to any one of claims 1 to 3, free or substantially free of Ciclosporin in any other form.

5. Ciclosporin according to any one of claims 1 to 4 in pure or 10 substantially pure form.

6. Ciclosporin according to any one of claims 1 to 5 in fine particulate form vherein the component particles have an average particle size of <200 yM.

7. Ciclosporin according to claim 6 vherein the component particles have an average particle size of from 0.5 to 200 yM.

8. Ciclosporin according to claim 6 vherein the component particles 20 have an average particle size of <25 yM.

9. Ciclosporin according to any one of claims 1 to 8 consisting^ predominantly, or consisting entirely or substantially entirely, of non-comminuted crystal material.

10. Ciclosporin according to any one of claims 6 to 9 vherein the component particles are comprised predominantly of, or are comprised entirely or substantially entirely of, non-comminuted micro-crystalline material.

11. Ciclosporin according to any one of claims 1 to 10 in sterile or substantially sterile condition. - 50

12. Process for the production of Cidosporin in non-solvate orthorhombic crystal form vhich process comprises crystallising Cidosporin from a Cidosporin non-solvate orthorhombic crystal bearing solvent medium and recovering Cidosporin non-solvate 5 orthorhombic crystals thus obtained.

13. A pharmaceutical composition comprising a) Cidosporin in non-solvate orthorhombic crystal form as active ingredient, together vith 10 b) a pharmaceutically acceptable diluent or carrier therefor.

14. A pharmaceutical composition according to claim 13 vherein Cidosporin present in said composition is predominantly, or is entirely or substantially entirely, in non-solvate orthorhombic 15. Crystal form.

15. A pharmaceutical composition according to claim 13 or 14 having at least one compartment or phase comprising Cidosporin in non-solvate orthorhombic crystal form, said crystal form being 20 distributed or distributable throughout said compartment or phase.

16. A pharmaceutical composition according to claim 15 vherein said crystal form is distributed or distributable homogeneously or substantially homogeneously throughout said compartment or phase.

17. A pharmaceutical composition according to any one of claims 13 to 16 comprising Cidosporin as defined in any one of claims 2 to 11.

18. A pharmaceutical composition according to any one of claims 13 to 30 17 in flovable form permitting topical application or application by injection or infusion. - 51

19. A pharmaceutical composition according to any one of claims 13 to 18 comprising from 0.05 to 30% by weight of Ciclosporin based on the total weight of the composition.

20. Ciclosporin according to any one of claims 1 to 11 for use as a pharmaceutical.

21. Ciclosporin according to claim 20 for use as an immunosuppressive or anti-inflammatory agent.

22. Ciclosporin according to any one of claims 1 to 11 for use in the preparation of a pharmaceutical composition for use as an immunosuppressive or anti-inflammatory agent.

23. Ciclosporin according to claim 22 for use in the preparation of a composition as defined in any one of claims 13 to 19.

24. The process for the production of ciclosporin in nori-solvate orthorhombic crystal form substantially as hereinbefore described with reference to Example 1.

25. The process for the production of ciclosporin in nori-solvate orthorhombic crystal form substantially as hereinbefore described with reference to Example 2.

26. Ciclosporin whenever produced by a process as claimed in claim 12.

27. Ciclosporin whenever produced by a process as claimed in claim 24.

28. Ciclosporin whenever produced by a process as claimed in claim 25.

29. A pharmaceutical composition according to any of claims 13 -19 substantially as hereinbefore described with reference to any of Examples 3 to 5. - 52

30. Ciclosporin in non-solvate orthorhombic crystal form substantially as hereinbefore described with reference to Example 1.

31. Ciclosporin in non-solvate orthorhombic crystal form substantially as hereinbefore described with reference to Example 2.