NO177228B - Process for Preparation of Ciclosporin in Orthorhombic Crystal Form - Google Patents

Description

The present invention relates to a method for producing ciclosporin in orthorhombic (P212121) crystal form. These and other features of the invention appear in the patent claims.

Ciclosporin, or cyclosporin A, is a known fungal metabolite with valuable pharmaceutical utility. Ciclosporin is a cyclic poly-N-methylated terminal decapetide of formula

wherein -MeBmt- represents an N-methyl-(4R)-4-but-2E-en-1-yl-4-methyl-(L)-threonyl residue.

Ciclosporin as well as methods for its preparation are e.g. described in US patent no. 4,117,118. The most important area of clinical investigation for ciclosporin has been as an immunosuppressive agent, particularly in connection with its use in the treatment of recipients of organ transplants, e.g. heart, lung, combined heart-lung, liver, kidney, pancreas, bone marrow, skin and corneal transplants and especially allogeneic organ transplants. Within this area ciclosporin has achieved considerable success and reputation and is now commonly used in treatment and is commercially available under the trade name SANDIMMUN<R-> or SANDIMUNNE<R>, e.g. in the form of an infusion concentrate, an oral solution or a gelatin-encapsulated solution, the latter two variants e.g. is described in US patent no. 4,388,307.

At the same time, investigations concerning the applicability of ciclosporin against various autoimmune diseases and inflammatory conditions, especially inflammatory conditions with an etiology comprising an autoimmune component such as arthritis, (e.g. rheumatoid arthritis, arthritis chronica progrediente and arthritis deformans) and rheumatic diseases, been intensive and reports and results in vitro, in animal models and clinical trials are widespread within the literature. Specific autoimmune diseases in which ciclosporin therapy has been proposed or used include autoimmune hematologic diseases (including, for example, hemolytic anemia, aplastic anemia, red cell anemia, and idiopathic thrombocytopenia), systemic lupus erythematosus, polychondritis, sclerodoma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, psoriasis, Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel disease (including, for example, ulcerative colitis and Crohn's disease), pemphigus, endocrinophthalmopathy, Graves' disease, sarcoidosis, multiple sclerosis, primary biliary cirrhosis , primary juvenile diabetes (diabetes mellitus type I), Behcef's disease, uveitis (anterior and posterior), vernal conjunctivitis, keratoconjunctivitis sicca, interstitial pulmonary fibrosis, psoriatic arthritis and glomerulonephritis (with and without nephrotic syndrome, e.g. extensive idiopathic nephrotic syndrome or minimal change nephropathy).

Further investigated areas have included potential use as an anti-parasitic agent, particularly an anti-protozoan agent, with possible applications including the treatment of malaria, coccidiomycosis and schistosomiasis, as well as use in the reversal of tumor resistance to chemotherapeutics, e.g. cytostatic therapy and to promote hair growth, e.g. in the treatment of alopecia.

In solid form, ciclosporin exists in both amorphous and crystalline form. In crystalline form, both a tetragonal and an orthorhombic modification are known.

The tetragonal modification (P4X), also referred to herein as "CY-A/X-I", is described together with methods for its preparation, in e.g. Austrian Patent No. 353,961. CY-A/X-I has a lattice a=b= 13.8 Å c= 41.2 Å, volume per asymmetric unit = 1974 Å<3>, a melting point at 140 - 150°C and comprises approximately 2 molecules of H20 per ciclosporin molecule.

A first orthorhombic modification (P212121) can be obtained by recrystallization 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 cyclosporine/di-isopropyl ether (ca. 1:2) solvate, with a crystal lattice of a = 12.5 Å, b = 22.9 Å, c = 28.4 Å, volume per asymmetric unit = 2027 Å<3>, and with a melting point at about 150°C. Orthorhombic ciclosporin solvate crystals can be obtained by recrystallization of supersaturated ciclosporin solutions in a suitable solvent over extended periods of time. Thus, the orthorhombic ciclosporin/di-isopropyl ether solvate modification prepared by Wenger is obtained by adding a concentrated solution of ciclosporin in methylene chloride (in which ciclosporin dissolves easily) to di-isopropyl ether (in which ciclosporin is somewhat less soluble) whereupon the methylene chloride is evaporated. Crystallization from the remaining diisopropyl ether then continues over a period of about 7 days at ambient temperature. Orthorhombic crystal forms comprising such ciclosporin solvates are also referred to herein generally as "CY-A/X-II".

A further orthorhombic (P212121) modification has now been identified which is characterized by the absence or substantially absence of any solvent component, i.e. consisting substantially of free ciclosporin.

The crystal lattice dimensions of the solvent-free or non-solvated orthorhombic ciclosporin modification are a =

12.7 Å, b = 15.7 Å, c = 3 6.3 Å, volume per asymmetric unit = 1804 Å<3>. The melting point is in the order of 180 to about 190°C or up to about 195°C, e.g. depending on the degree of purity/source as indicated in Examples 1 and 2 below, where specific methods of achieving this modification are described. Ciclosporin in non-solvated (or solvent-free) orthorhombic crystal form is also referred to herein as "CY-A/X-III". Additional data for the characterization of CY-A/X-III (X-ray diffraction) are given in Table III and Figs. IV below, together with corresponding data for CY-A/X-I and CY-A/X-II.

Despite the significant success and reputation that ciclosporin has achieved and despite the important contribution to medical research, especially in the field of organ transplantation, which has already been carried out, its application in practice is still not ideal and problem-free. A particular area of concern is the provision of galenic forms which allow appropriate or completely satisfactory delivery, or which are best suited for the treatment of specific diseases or conditions where ciclosporin has already been identified as a potential drug of choice.

These problems can partly be attributed to ciclosporin's special physical properties, e.g. its relative limited solubility or (heretofore experienced) compatibility with available pharmaceutically usable carrier media. Such problems include both the nature of some of the disease states for which ciclosporin represents a potential first-line therapy, as well as the presence of unwanted side effects that ciclosporin can induce when administered orally. Ciclosporin has e.g. has been identified to be effective in the treatment of psoriasis by oral administration. Psoriasis is a widespread condition that causes considerable suffering in the patient and for which there is so far no generally accepted, effective therapy. Those suffering from psoriasis clearly represent a patient population in need of therapy. While the available evidence would suggest that ciclosporin would be able to accommodate this, it is unlikely that oral ciclosporin therapy would find acceptance as a common agent for 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 delivery of ciclosporin, including galenic forms 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 or other dermatological diseases and conditions, e.g. atopic dermatosis and alopecia, or for the treatment of uveitis. However, such systems have so far not provided any noticeable benefits or have not proved completely satisfactory.

Thus, various reported attempts to treat individuals with psoriasis by applying the known oral solution directly to the surface site of the psoriatic lesion have yielded little or no success, and success with individuals suffering from atopic dermatosis is severely limited. Likewise, while topical application to the eye has been found to be highly effective as a means of preventing e.g. corneal transplant rejection, or for the treatment of diseases in the anterior segment of the eye, no topical preparation has so far been reported that would be able to provide a quick, safe and effective treatment of diseases or conditions that affect places that sit deeper inside the eye, e.g. for general treatment of uveitis, including posterior uveitis.

Similar problems have been encountered in connection with the development of other ciclosporin drug delivery systems, e.g. the injectable forms, such as is suitable for intra-articular injection for the local treatment of arthritic diseases, such as e.g. mentioned above, or for intralesional injection, e.g. for the treatment of a severe psoriatic lesion, or alternatively oral delivery systems such as e.g. have modified bioavailability or drug release properties.

Attempts so far carried out to form ciclosporin preparations have made use of the compound in solution or, less commonly, in amorphous or CY-A/X-I form, the latter e.g. is the crystal form identified for e.g. drug registration purposes. Despite the considerable efforts made to find new or improved alternatives, the oral solution and infusion concentrate already discussed remain the only widely used delivery systems for ciclosporin.

It has now surprisingly been found that orthorhombic crystalline forms of ciclosporin, e.g. CY-A/X-II and especially CY-A/X-III, are of particular advantage and use in the production of ciclosporin galenic preparations of various types.

It has been found in particular that such crystal modifications can be used for the production of galenic preparations comprising ciclosporin in the form of stable, fine particles and/or with increased stability or better release properties. This has opened the way for the production of new and improved ciclosporin drug delivery systems, e.g. with sustained or delayed ciclosporin delivery and/or which are adapted to or appropriate for the treatment of diseases or conditions which have so far been unresponsive to ciclosporin therapy or where usual ciclosporin therapy has naturally been less appropriate. More particularly, it has been found that when using such crystal modifications, e.g. in the form of fine particles as mentioned above, galenic forms can be obtained which are suitable for topical or dermal application or for topical ophthalmic application, for the treatment of diseases or conditions affecting the skin or eyes, e.g. as mentioned above, and in particular for the treatment of psoriasis, atopic dermatitis, contact dermatitis, ocular symptoms of Behchef's disease, uveitis and keratoconjunctivitis sicca. Application of such crystal modifications also makes it possible to produce injectable forms of ciclosporin with sustained or delayed release properties and which are thus of particular use, e.g. in the treatment of diseases or conditions where the required frequency of administration may otherwise inhibit the applicability, e.g. in the treatment of arthritis or a similar disease in the joints by intra-articular injection or for the treatment of psoriasis or similar diseases of the skin by intralesional injection.

For such purposes, orthorhombic crystal forms, particularly Cy-A/X-III, have been found to be much better suited than other available forms, particularly amorphous ciclosporin or Cy-A/X-I, e.g. with regard to physico-chemical properties, especially stability, release properties, i.e. release of ciclosporin from the crystal lattice, and its susceptibility to the necessary procedures regarding galenic preparations. Cy-A/X-III is a new and orthorhombic crystal form for galenic and/or therapeutic use, where

A is CY-A/X-III, i.e. ciclosporin in a non-solvated orthorhombic crystal form;

especially:

A<1> CY-A/X-III with a crystal lattice or other physical properties as essentially defined in the foregoing,

and or

A<2> CY-A/X-III with X-ray diffraction properties which are essentially given in Table III or which are shown in fig.

IV.

The present invention provides a process for the preparation of ciclosporin in orthorhombic (P2^2 ^2],) crystal form with a crystal lattice of dimensions a = 12.7 Å, b = 15.7 Å, c = 3 6.3 Å, vol. per asymmetric unit = 1804 which is characterized in that ciclosporin is dissolved in a solvent medium comprising a high molecular weight polymer ether, at a temperature of more than 40°C, ciclosporin is crystallized from said solvent medium bearing ciclosporin in orthorhombic (P212121) crystal form as indicated above and the crystal form thus obtained is recovered.

Suitable CY-A/X-III-bearing media for use in the above-mentioned method is, as mentioned, a solvent medium for ciclosporin comprising a polymer ether of high molecular weight, e.g. with a molecular weight >200. Examples of suitable polymer ethers are polyethylene and polypropylene glycols, e.g. with molecular weight >200; glycerin polyethylene glycol esters, e.g. glycerin polyethylene glycol ricinoleates and glycerin polyethylene glycol oxystearates such as are available under the trade name "Cremophor EL" and Cremophor RH" (see Fiedler, "Lexikon der Hilfstoffe" 2nd edition, Vol 1, pages 257 and 258); polyoxyethylene sorbitan esters such as are available under the trade name "Tween" (see Fiedler, 2, pages 972 - 975); polyoxyethylene esters, e.g., polyoxyethylene stearic acid esters, e.g., as available under the trade name "Myrj" (see Fiedler, 2, page 636); and transesterification products of natural-oil triglycerides and polyalkylene polyols, including transesterification products of corn oil, kernel oil, almond oil, peanut oil, olive oil, palm oil and mixtures thereof with polyethylene glycols, in particular polyethylene glycols with a molecular weight from 200 - 800, for example available under the trade name "Labrafil" (cf Fiedler, page 539). Particularly suitable are media including polyethylene glycols with a molecular weight of 200 to 600, especially of 300 to 400.

As will be understood, the CY-A/X-III carrying medium may comprise one or more components in addition to those given above, e.g. lower molecular weight alcohols and/or water, as a means of modifying solvent/crystallization properties. A particularly suitable CY-A/X-III carrying medium has been found to be one comprising about 0.5 volume ethanol, about 8.0 volume polyethylene glycol (300) and about 1.0 volume H 2 O.

The method according to the present invention comprises that ciclosporin, e.g. in amorphous or tetragonal form, is first dissolved in the selected CY-A/X-III-bearing medium at an elevated temperature. The dissolution temperature will of course vary depending on the particular medium chosen, but is generally > 40°C and more particularly > 50°C. In the case of a polyethylene glycol-based medium, the dissolution temperature will generally be > 70°C, e.g. from 75 to 130°C. The solution obtained will suitably comprise more than 5%, preferably more than 10%, e.g. up to 40-60%, preferably up to a maximum of about 20% by weight ciclosporin based on the total weight of the solution. Crystallization of ciclosporin then takes place with suitable cooling over a relatively extended period of time, e.g. in the order of 10 - 35 minutes or more, depending on the desired crystal growth.

If desired, nucleation procedures can be initiated, e.g. using sound processing or kimt il sentence. Where ethanol/polyethylene glycol/H20 crystal-bearing media, e.g. as described above, is used, one has e.g. found that nucleation can be easily initiated at temperatures of 60 - 90°C, e.g. 65 - 85°C using sound vibrations of approximately 20,000 cycles per second. As indicated in example 2 below, by continuous sound treatment in this way during the entire crystallization process, microcrystals with a very small and relatively constant particle size can be obtained.

The rate of crystal growth and the size of the crystals obtained will of course depend on the particular medium chosen and on the conditions under which the crystallization is carried out, e.g. whether sound processing is used or not and at what frequency. For the production of e.g. galenic preparations which are described in the following, CY-A/X-III will preferably be used in the form of small particles, e.g. with an average particle size of < 200 \ Lm, e.g. from 0.5 to 200 Jim. Where the initially obtained crystals have a relatively large size, particulate preparations of the desired size can of course be obtained by commonly used crushing procedures, e.g. by grinding, crushing, pulverizing, micronizing or otherwise fragmenting to a reduced size. While particulate preparations obtained in this way may be perfectly suitable for some purposes, e.g. for the preparation of topical preparations for application to the skin, the use of crushing or equivalent techniques will in many cases not be preferred. The reasons for this are that the products from such procedures have a tendency to exhibit a relatively large particle size distribution and have a relatively coarse or abrasive structure as a consequence of crystal breakage. In addition, crushing will tend to destroy the physical properties of the natural crystal surface and is in all cases difficult or expensive to implement, while required conditions in terms of sterility, e.g. in the manufacture of galenic preparations for intra-articular administration, is maintained. Because of. it is usually preferred to carry out the method according to the invention in such a way that one obtains crystals, e.g. microcrystals, with dimensions within the desired particle size range. Thus, for use in the production of e.g. preparations for intra-articular or other parenteral administration, the use of CY-A/X-III particle preparations in an unbroken microcrysalline state (ie, CY-A/X-III microcrystals in a native or non-degraded state, e.g., which are not subjected to crushing, grinding, micronization or other reducing procedures) is preferred.

In order to obtain CY-A/X-III microcrystals which can be used directly in an unbroken state in the above-mentioned manner, the method according to the invention will suitably be carried out with stirring, e.g. 200 - 300 r.p.m. which is increased to e.g. 700 r.p.m. during crystallization and especially when using ultrasonic vibration, e.g. within the range from 10,000 to 30,000 cycles per second (c.p.s.), suitably on the order of about 20,000 cycles per second.

By varying the CY-A/X-III carrying medium and adjusting the cooling rate and degree of agitation during the recrystallization procedure in accordance with known techniques and as exemplified hereinafter, CY-A/X-III microcrystals with varying average particle size, e.g. as indicated above or as described below for use in connection with individual types of galenic preparations.

As indicated above, crystallization in accordance with the method of the present invention will conveniently be carried out under sterile conditions. Purification of the obtained CY-A/X-III can be carried out in the usual way, e.g. by cleaning with polyethylene glycol (300): water e.g. in a ratio of 3 - 4:1 parts by weight/washing with warm water, as described in the examples. Obtained CY-A/X-III will conveniently be recovered without and substantially without ciclosporin in any other form, preferably in pure or substantially pure form. The purification step as indicated above serves to prevent the formation of amorphous ciclosporin.

Ciclosporin produced according to the invention can also be in the form of: A<3> : CY-A/X-III according to any of the above A-A2 without or essentially without ciclosporin in any other form, e.g. without or substantially without

ciclosporin in amorphous form of CY-A/X-I or of CY-A/X-II; A4 : CY-A/X-III according to any of the above

A - A3 in pure or substantially pure form;

A5 : CY-A/X-III according to any of the above A - A<4> in the form of fine particles, e.g. with particle size as given above or as described below, e.g. with regard to galenic forms

comprehensive CY-A/X-III;

A<6> : CY-A/X-III according to any of the above A-A<5> comprising mainly or wholly or substantially wholly non-pulverized crystalline material, e.g. CY-A/X-III according to the above A<5> where the component particles comprise mainly or entirely or in total

substantially all non-pulverized microcrystals; and

A<7> : CY-A/X-III according to any of the above A - A<6> in sterile or essentially sterile condition, e.g. in a state suitable for pharmaceutical applications, particularly suitable for parenteral administration, e.g. administration by parenteral injection such as e.g. intra-articular injection.

Ciclosporin may be present in preparations in any suitable orthorhombic crystal form, e.g. as CY-A/X-II or CY-A/X-III. Application of CY-A/X-II will, however, generally be less preferred due to its physicochemical properties which are less suitable for use both in terms of tolerability and applicability for the production of effective ciclosporin-based pharmaceutical preparations. Most preferably, ciclosporin will be present in preparations such as CY-A/X-III, e.g. according to any of the above A-A<7>.

Although ciclosporin is present in a preparation in solid (crystalline) form, the preparation itself may be of any suitable nature including solid forms such as capsules, tablets, coated tablets, powders and granules, semi-solid forms such as ointments, gels , creams and pastes, as well as liquid forms comprising e.g. the defined ciclosporin component in the form of fine particles

(e.g. in micronized form or in the form of microparticles)

suspended or dispersed in a liquid pharmaceutical diluent or carrier in which the selected crystal form can be retained.

The preparation includes both systems that comprise a single ciclosporin-containing phase, or rooms e.g. simple creams, gels, ointments, etc. as mentioned above or simple one-compartment capsules or ampoules, e.g. capsules of hard or soft gelatin or ampoules for injection as well as systems comprising a number of phases or compartments. Thus, the preparation can e.g. comprise a ciclosporin-containing phase or compartment together with one or more non-ciclosporin-containing phases or compartments, e.g. coating, carrier, retarding phases or placebo phases, or phases or compartments comprising other medicinal substances or excipients. As preparations, there may be mentioned coated tablets with a core comprising ciclosporin in orthorhombic crystal form surrounded by e.g. non-ciclosporin-containing coating phases or the like; coated tablets comprising a core or coating comprising ciclosporin in orthorhombic form and with a non-ciclosporin containing coating or core; mixing granules comprising a first granule phase comprising ciclosporin in orthorhombic crystal form and a second non-ciclosporin-containing granule phase; and divided capsules with two or more compartments, e.g. divided by means of an inner wall, one compartment containing ciclosporin in orthorhombic form and one compartment not containing ciclosporin. Other options and variants will be clearly apparent to a specialist in the field. Ciclosporin is preferably present in a preparation mainly or preferably entirely or entirely in orthorhombic crystal form, e.g. as CY-A/X-III.

A preparation may suitably comprise at least one phase or compartment comprising ciclosporin in orthorhombic crystal form, e.g. CY-A/X-III, through which said crystal form is evenly distributed. Such systems include e.g. homogeneous creams, ointments, gels and the like as well as liquid systems in which contained crystalline ciclosporin is or can be easily distributed homogeneously, e.g. using manual stirring.

A preparation can have forms that are suitable for topical application, e.g. dermal, or topical ophthalmic application; for parenteral administration, e.g. by infusion or injection including subcutaneous or intramuscular injection and especially as intralesional or intra-articular injection; as well as forms of enteral delivery, e.g. as suppositories, pessaries and the like and as oral dosage forms. Enteral forms, e.g. however, oral dosage forms would be less preferred.

Preferably, orthorhombic crystalline ciclosporin, e.g. CY-A/X-III be present in a preparation in the form of fine particles, e.g. in micronized form, in microparticulate form or in non-pulverized microcrystalline form. Appropriate particle size will of course vary depending on the type of preparation, the preparation's intended use and method of administration and the desired effect. In general, however, the average particle size will suitably be in the range of 0.5 to 200 µm. Particle size ranges in e.g. connection with the manufacture of oral, topical or injectable forms is further discussed below.

Topical forms, e.g., for dermal or topical ophthalmic application, e.g., for the treatment of psoriasis or atopic dermatosis or of diseases or conditions in practice that require immunosuppressive therapy, e.g. uveitis, conjunctivitis, extensive keratoconjunctivitis sicca or vernal keratoconjunctivitis, or cornea graft.

Gels, creams, ointments and the like for topical or dermal application can be prepared in accordance with any of the generally used known techniques, e.g. by mixing ciclosporin in orthorhombic crystal form with a suitable flowable base material, e.g. an aqueous gel, e.g. an aqueous cellulose ether-based gel, with the addition of suitable surfactants, thickeners, etc. comprising e.g. aerosil.

The ciclosporin component is preferably present in the form of fine particles, e.g. with an average particle size from 0.5 to 200 µm. The average particle size is preferably less than 60 - 80 µm, more preferably less than 40 µm, and even more preferably less than 30 µm, and most preferably less than 20 µm. The particles will advantageously have as narrow a particle size distribution as possible. Preferably, essentially all particles present will be smaller than 60 - 80 (lm.

A preparation for topical and ophthalmic use, i.e. for application to the eye, may include e.g. suitably thickened suspensions or dispersions, e.g. as presented in the form of viscous eye drop preparations or creams or gels for ocular application. In the case of preparations to be applied to the eye, the average particle size of the crystalline ciclosporin material present will suitably be less than 25 µm. More preferably, substantially all of the particles present will be smaller than 25 µm. The average particle size of such eye drop preparations will suitably be in the range from 0.1, or preferably from 0.5 to 20 µm or preferably up to 10 µm. The hydrophilic/lipophilic content of the carrier system for use in forms of dermal application or for use in preparations to be applied to the eye may vary depending on the particular surface to which the preparation is to be applied, eg depending on the characteristics of the particular skin area to be treated.

A preparation for topical, especially dermal application suitably comprises a carrier medium in which the active ingredient (ciclosporin component) is only slightly soluble. The solubility of the ciclosporin component (eg CY-A/X-III) in such a preparation is suitably less than 10%, preferably less than 1.0% by weight. More preferably, the solubility is less than 0.1% and most preferably less than 0.001% by weight.

Suitable carrier media include hydrophilic, water-miscible carriers as well as hydrophobic carriers. Hydrophilic water-miscible carrier components that are suitable are those of relatively low volatility. Examples of suitable carrier components belonging to this group include the groups polyalkylene glycols, propylene glycol, glycerol and ethylene glycol. Suitable polyalkylene glycols include in particular liquid polyethylene glycols, e.g. those with an average molecular weight up to about 600, e.g. with an average molecular weight of from 250 to 450, especially about 300.

Representative preparations for e.g. topical application therefore comprises: C<1> a) Ciclosporin in orthorhombic, preferably non-solvated orthorhombic crystal form, e.g. CY-A/X-III, e.g. as defined above; b) A hydrophilic, water-miscible carrier, e.g. as described above; and eventually

c) Water of pharmaceutical quality.

The amount of component (a) present will suitably be i

order of magnitude from 0.1 to 30.0%, in particular from 0.5 or 1.0 to 20%, e.g. about 1.0, 2.0, 5.0 or 10.0% based on the total weight of the mixture. Components (b) and (c) will suitably be present in a ratio of from 1:0 to 1:20 expressed in parts by weight. More appropriate would be components (b) and (c).

present in a ratio of 1:0.5 to 1:5, e.g. from 1:1 to 1:3 and most suitably from 1:1.5 to 1:2.5 expressed in parts by weight.

Such preparations will suitably further include:

d) a surfactant.

Suitable components (d) include, among others:

d<1> Oesterification products of triglycerides and poly alkylene polyols from natural vegetable oils. Such transesterification products are known from the art and can e.g. is achieved in accordance with the general procedures described in US Patent No. 3,288,824. They include transesterification products from various natural vegetable oils (e.g. non-hydrogenated), such as corn oil, kernel oil, almond oil, peanut oil, olive oil and palm oil and mixtures thereof with polyethylene glycols, in particular polyethylene glycols with an average molecular weight from 200 to 800. Products obtained by transesterification of two molar parts of triglyceride from natural vegetable oil with one molar part of polyethylene glycol are preferred (e.g. eg with an average molecular weight from 200 to 800). Various forms of transesterification products from the defined group are known and commercially available under the trade name Labrafil [see Fiedler, "Lexikol der Hilfstoffe", 2nd revised and expanded edition (1981), volume 2, page 539], Particularly applicable as components in the preparations are the products: Labrafil M 1944 CS, a transesterification product of kernel oil and polyethylene glycol with an acid number of approximately 2, a saponification number of 145 - 175 and an iodine number of 60 - 90; and Labrafil M 2130 CS, a transesterification product of a C12 - C18 glyceride and polyethylene glycol with a melting point of 35 - 40°C, an acid number < 2, a saponification number of 185 - 200 and

an iodine number < 3.

d<2>Reaction products of a natural or hydrogenated castor oil and ethylene oxide. Such products can be obtained in a known manner, e.g. by reacting natural hydrogenated castor oils with ethylene oxide, e.g. in a molar ratio from

1:35 to 1:60, with possible removal of the polyethylene glycol components from the product, e.g. in accordance with the methods given in German specification Nos. 1,182,388 and 1,518,819. Particularly suitable are the various surfactants available under the trade name Cremophor. Particularly suitable are the products Cremophor RH 40 with a saponification number of 50 - 60, an acid number < 1 and an iodine number < 1, a water content (Fischer) < 2%, nd<60> of 1.453 - 1.4578 and HLB of 14 - 16 ;

Cremophor RH60 with a saponification number of 40 - 50, an acid number < 1, an iodine number < 1, a water content (Fischer) of 4.5 - 5.5%, nd<25> of 1.453 - 1.57 and HLB of 15 - 17; and Cremphore EL having a molecular weight (by vapor osmometry) of about 1630, a saponification number of about 65 - 70, an acid number of about 2, an iodine number of about 28 - 32 and an nd<25> of about 1.471. Also suitable for use in this category are the various surfactants available under the trade name Nikkol, e.g. Nikkol HCO-60. Nikkol HCO-60 is a reaction product of hydrogenated castor oil and

ethylene oxide with the following properties: acid number 0.3;

saponification figure 47.4; hydroxy value 42.5, pH (5%) 4.6;

color APHA = 40; melting point = 36.0°C; freezing point =

32.4°C; H 2 O content (%, KF) = 0.03.

d<3>Polyoxyethylene sorbitan fatty acid esters or polysorbates,

e.g. of the type that is known and commercially available under e.g. trade name Tween and Armotan (see Fiedler, 2, pages 745 - 746 and 972 - 975) including the products Tween

20- [polyoxyethylene-(20)-sorbitan monolaurate], 40-[polyoxyethylene-(20)-sorbitan monopalmitate], 60- [polyoxyethylene-(20)-sorbitan monostearate], 65-[polyoxyethylene-(20)-sorbitan tristearate], 80- [polyoxyethylene-(20)-sorbitan monooleate], 85-[polyoxyethylene-(20)-sorbitan trioleate], 21- [polyoxyethylene-(4)-sorbitan monolaurate], 61- [polyoxyethylene-(4)-sorbitan monostearate), and 81- [ polyoxyethylene-(5)-sorbitan monooleate].

d<4>Polyoxyethylene fatty acid esters, e.g. polyoxyethylene stearic acid esters of the type known and commercially available under the trade name Myrj (see Fiedler, 2, page 636) as well as polyoxyethylene fatty acid esters known and commercially available under the trade name Cetiol HE (see Fiedler, 1, page 228), and polyoxyethylene- fatty alcohol ethers, e.g. polyoxyethylene stearyl ether, oleyl ether or cetyl ether, e.g. of the type known under the trade name Brij (see Fiedler, 1, 184 - 186), e.g. Brij 78 and 96 and Cetomacrogel 1000 (see Fiedler, 1,

229).

d<5> Polyoxyethylene-polyoxypropylene copolymers, e.g. of the type known and commercially available under the trade names Pluronic and Emkalyx (see Fiedler, 2, page

720 - 722).

d<6>Sorbitan fatty acid esters e.g. of the type known and commercially available under the trade name Span, e.g. comprising sorbitan monolauryl, -monopalmityl,

-monostearyl, -tristearyl, -monooleyl and -trioleyl esters

(see Fiedler, 2, pages 850 - 851).

d<7> Partial glycerides, e.g. products comprising mono- and/or di-glyceride components, e.g. mixed mono-/di-/triglyceride/free glycerol based surfactants as well as e.g. glycerol monostearates and glycerol monooleates. Examples of such products include those that are known and commercially available under the trade name Imwitor (see Fiedler, 1, 491) e.g. Imwitor 191 and Imwitor 780 and Softigen (see Fiedler, 2,

833) e.g. Softigen 701.

d<8>Ionic surfactants and emulsifiers such

such as sodium lauryl sulfate and sodium cetostearyl sulfate.

Components (d), when present, are suitably present in an amount of up to a maximum of 30%, more preferably a maximum of 15% based on the total weight of the composition. The most suitable components (d) are present in an amount from 0.5 to 10%, e.g. approximately 1.0%, based on the total weight of the preparation.

Such preparations will also suitably include:

e) A thickening agent.

Suitable component(s) include, for example:

e<1> Polymethyl acrylate resins, e.g. that type that is known and commercially available under the trade name

Eudispert (see Fiedler, 1, 371 - 372).

e<2> Cellulose derivatives comprising e.g. ethyl, propyl,

methyl, hydroxypropylmethyl and carboxymethyl cellulose.

e<3> Polyvinyl resins, e.g. including polyvinyl alcohols and polyvinyl pyrrolidones, as well as other polymeric materials including gelatin, alginates, pectins, tragacanth gum, gum arabic, xanthan gum, and polyacrylic acids known and available under the trade name

Carbopol (see Fiedler, 1, 206 - 107).

e<4> Materials such as silica gel, bentonite and magnesium aluminum silicate.

Component(s), when present, are suitably present in an amount up to 20%, more preferably up to 10%, based on the total weight of the composition. Most suitably, the component(s) are present in an amount of from 0.5 to 15%, e.g. from 1.0 to 3.0%, based on the total weight of the preparation.

In addition, such preparations may also suitably comprise an antimicrobial agent such as methylparaben, propylparaben, benzalkonium chloride or benzyl alcohol, e.g. in an amount from 0.05 to 0.5% or in the case of benzyl alcohol, up to 2.0%, e.g. about 0.1% or in the case of benzyl alcohol, about 1.0% by weight based on the total weight of the preparation.

Alternative carrier media include hydrophobic materials, e.g. those that meet the solubility criteria with respect to the active ingredient described above. Further preferred preparations for e.g. topical application, therefore comprises: c<2> a) Ciclosporin in orthorhombic, preferably non-solvated orthorhombic crystal form, e.g. CY-A/X-III, e.g. as defined under any of the above A - A<7>; and

f) A hydrophobic carrier, e.g. as indicated above.

The amount of component (a) present will suitably be as

described above in connection with the preparations defined under C<1>. Suitable components (f) include e.g. petroleum derivatives, including mineral oils, fats and gels, e.g. petrolatum (mineral fat or petroleum jelly), paraffin oil (white mineral oil or liquid paraffin), or any of the products known and commercially available under the trade name Vaseline [see Fiedler loe. one. 2, 986], as well as nonionic lanolin-based derivatives available under the trade name Amerchol, e.g. Amerchol CAB [Fiedler, 1, 119 - 120].

Further suitable components (f) include e.g. natural or saturated vegetable oils and fats, e.g. fractionated coconut oil known and available under the trade name Miglyol, e.g. Miglyol 812 (see Fiedler, 2, 616) and peanut and saturated peanut oils, as well as higher fatty alcohols or branched alcohols and fatty acid esters, such as stearic alcohol, cetyl palmitate and 2-octyldodecanol known and commercially available under the trade name Eutanol G (see Fiedler, 1, 375).

Such preparations may also comprise one or more of the components (d) and (e) described above, as well as one or more antimicrobial agents, e.g. as described above.

Such preparations may in particular further comprise one or more components (d), suitably in an amount up to at most 30%, more preferably at most 10%, and suitably in an amount from 1 to 15% by weight, based on the total weight of the preparation. Hydrophobic-based preparations in accordance with C<2> and comprising an emulsifier (d), can also be prepared in the form of creams or emulsions by further addition of water. The resulting creams or emulsions can either be of the oil-in-water or water-in-oil type. The amount of water present in such preparations can thus vary from e.g. 5 to 80%, based on the total weight of the preparation, suitably from 20 to 70%.

The amount of component(s)/antimicrobial agent, when incorporated, will be of the same or similar order of magnitude as those given in connection with the preparations according to C1 above.

The amount of any topical form applied will, of course, vary depending on the concentration of the preparation, the condition to be treated and the effect desired, as well as, in the case of dermal application, the area to be covered. In the case of preparations for dermal application, e.g. in the treatment of psoriasis or atopic dermatosis, it will generally be sufficient to apply an amount which provides a ciclosporin dosage in the order of 0.1 to 5 mg/cm<2>, e.g. approximately 0.5 mg/cm<2>, when applied, for example, twice a day to the desired location. In the case of ophthalmic preparations, administration can conveniently be carried out using preparations comprising from 0.1 to 10%, e.g.

1.0 or 2.0% by weight ciclosporin, added in drop form to each eye 1-3 times a day.

Injectable forms, e.g., for subcutaneous, intramuscular or other parenteral injection, including especially intra-articular injection, e.g., in the treatment of arthritic diseases such as rheumatoid arthritis, or for intralesional injection, e.g., in the treatment of psoriasis.

Dispersions, suspensions, emulsions and similar preparations suitable for injection may be prepared in accordance with any of the known techniques generally employed, e.g. by dispersing, suspending or otherwise distributing ciclosporin as orthorhombic crystals in the form of fine particles in a suitable liquid carrier medium with the addition of suitable emulsifiers, surfactants, stabilizers or flocculating agents, etc. Preferably, the ciclosporin components in such preparations will have an average particle size of < 100 µM, more preferably < 50 µM and most preferably < 20 µM. The average particle size will suitably be in the order of 0.1 or 0.5 up to the indicated maximum of 20, 50 or 100 µM.

Where injectable forms are desired, e.g. for subcutaneous or intramuscular injection, to achieve e.g. a depot effect, the use of a larger particle size will be possible. Where forms of intralesional administration or especially intra-articular injection are desired, the use of preparations with smaller particles will be indicated. For such applications, suitable average particle size is e.g. in the order of 0.1 or 0.5 to 20 |1M, e.g. from 5 to 15 |1M.

To obtain stable particles in the injectable forms, e.g. to avoid the size of larger crystalline particles increasing at the expense of smaller particles, the particles will preferably have as uniform a size as possible. Appropriate variation between maximum and minimum particle size will not exceed about 50 µM, more preferably about 20 (IM.

Although the desired particle size can be achieved using suitable pulverization procedures, e.g. micronisation, to minimize particle size variations, to avoid the presence of fragmented particles, to avoid modifications of the crystal face, e.g. in order to retain the moisture properties, and to more easily meet the requirements regarding sterility, it will generally be preferred to use non-powdered microcrystals, i.e. crystals that have been grown to the desired size as described above.

Injectable preparations thus suitably include:

C<3> a) Ciclosporin in fine particle shape orthorhombic,

preferred non-solvate orthorhombic crystal form, e.g. CY-A/X-III, e.g. as indicated under any of the above A-A<7>, particularly A<6> and A<7>, which is or may be distributed in

b) an injectable carrier medium.

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

quality.

In addition, the preparation as indicated under C<3> will suitably include: g) a surface-active agent such as a dispersant or emulsifier and/or a flocculating agent, to effect or maintain distribution of the particles in the carrier phase.

Suitable surfactant components (g) include polyoxyethylene sorbitan fatty acid esters or polysorbates, e.g. as given under (d3) above, e.g. Tween 80 or Polysorbate 80 (see Fiedler, 2, page 746). Suitable flocculants (g) include ethylenediaminetetraacetate and salts thereof, e.g. Calcium EDTA and Sodium EDTA, as well as other equivalent chelating agents and peptizing agents such as succinic acid or citric acid.

To maintain the particle size distribution, a stabilizing agent is also suitably incorporated. Suitable stabilizers include e.g. gelatins or modified gelatins such as plasma expander known and commercially available under the trade name Gelafundin and Haemaccel or gelatins soluble in cold water of very pure collagen hydrolysates.

Preparations in accordance with the above-mentioned C<3> may also comprise one or more antimicrobial agents, e.g. as described above, as well as peptizing agents such as succinic or citric acids.

The ratio between the components (a):(b) in the preparations as defined under C<3> is suitable in the order of magnitude from 1:10 to 1:1,000, preferably in the order of magnitude from 1:50 to 1:200 e.g. approximately 1,100, expressed in parts by weight.

In order not to unduly disturb the particle stability, surfactants present as component (g) will preferably be present in only minor concentrations, e.g. in the order of 0.1 to 2.0%, suitably about 1% by weight, based on the weight of component (a). Where a flocculant is present, the ratio of flocculant to component (a) is suitably in the order of 1:3 to 1:8, e.g. approximately 1:5, expressed in parts by weight. When a stabilizing agent is present, the ratio of component (a) to the stabilizing agent is suitably in the order of 1:5 to 1:30, more preferably 1:10 to 1:30, e.g. approximately 1:20, expressed in parts by weight. When a peptizing agent is present, the ratio of the peptizing agent to component (a) is suitably in the order of 1:1 to 1:10, e.g. approximately 1:15, expressed in parts by weight.

The amount of ciclosporin present in unit dose forms for injection will of course vary depending on e.g. the condition to be treated, the injection site and the desired effect, e.g. whether a depot effect or a relatively rapid release into the surrounding tissues is expected. Suitable individual unit dosage forms will comprise from 10 to 500 mg, particularly from 20 to 100 mg ciclosporin/dose.

Unit dosage forms may also include more complex systems, e.g. syringes or two-compartment syringe containers in which particulate ciclosporin together with other suitable components, e.g. stabilizing agent or antimicrobial agent, is kept in a first chamber and the carrier medium is kept in a second chamber, the components of the two chambers being brought together into a mixture, e.g. after activation of the syringe plunger. Such spray tanks with double chambers are well known in the state of the art.

Oral dosage forms, for example, for the treatment of autoimmune and other diseases and conditions as given above, for example, to prevent transplant rejection.

Oral dosage forms can be prepared using any suitable carrier or carrier medium known and used in the art. Liquid or semi-solid oral dosage forms can e.g. include ciclosporin in orthorhombic crystal form, e.g. such as CY-A/X-III, together with pharmaceutically acceptable diluents or carriers in which the crystal form is insoluble or substantially insoluble. Such carriers and diluents include e.g. transesterification products of triglycerides and polyalkylene polyols from natural vegetable oil as described above under (d<1>), e.g. Labrafils as well as reaction products of natural or hydrogenated castor oil and ethylene oxide as described above under (d<2>), especially liquid Cremophore products, e.g. Cremophore EL.

Ciclosporin is preferably present in the form of fine particles, e.g. with an average particle size of from 0.5 to 200 µm, in particular in the order of magnitude from 0.5 to 30 µM, and preferably less than 20 µM, e.g. from 0.5 to 10 p.M.

For suitable delivery, the preparations as mentioned above are suitably filled in capsules of hard or soft gelatin.

Unit dosage forms for oral administration suitably comprise from 25 to 75, or up to 200, e.g. approximately 50 or 100 mg ciclosporin/unit dose. Such forms are supplied e.g. from 1 to 4 times daily to achieve individual ciclosporin concentrations in the patient's serum (e.g. as determined by RIA) of the same or equivalent order of magnitude as those achieved using standard ciclosporin therapy, e.g. using an available ciclosporin solution for oral administration.

Further galenic forms of delivery by routes which are equivalent or alternative to those described above will be clear to a person skilled in the art.

Example 1

Preparation of CY-A/X-III

a) 100 g ciclosporin in amorphous form is dissolved with stirring in 400 g polyethylene glycol 300 at 50°C. Development of CY-A/X-III starts shortly after dissolution. The solution is allowed to stand for 24 - 48 hours at 35 - 40°C to end crystallization and is then diluted with 1000 ml of water with vigorous stirring. The precipitate obtained is separated by filtration, washed with H 2 O to remove residual polyethylene glycol and dried to give CY-A/X-III in essentially pure form: mp = 195°C, particle size =

100 - 200 JIM.

CY-A/X-III forms can be prepared analogously to the above procedure by using the following materials as solvent instead of polyethylene glycol 300.

In each case, the average particle size has been obtained in the order of 50 to 200 (IM. Determined lattice structure is as defined above. X-ray diffraction data is as described below with reference to table III and fig. IV.

Example 2

Preparation of CY-A/X-III

20 g of ciclosporin as CY-A/X-I are dissolved in 60 ml of ethanol and condensed to 1/6 of the initial volume by distillation under reduced pressure at more than 60°C. 160 ml of ethylene glycol (molecular weight 300) + 20 ml of H20 are then added slowly at 50 - 85°C with stirring at 300 r.p.m. An ultrasonic vibrator/mixer is installed and operates at 20,000 c.p.s. Frost formation is initiated at 65 - 75°C over 35 minutes. The obtained dispersion is then cooled linearly over 10 - 20 minutes to 40°C with continuous stirring at 700 r.p.m. and ultrasonic vibration. The resulting thick pasty mixture is filtered, purified with polyethylene glycol (molecular weight 300)/H 2 O (3:1 v/v), washed with hot water (30°C) and dried at 50°C under high vacuum to give CY-A/X -III in pure form: mp = 192°C. Particle size = 3-15 jiM. Determined lattice structure is as defined above. X-ray diffraction data are described below with reference to table III and fig. IV.

Repeated experiments indicate that the crystal size is influenced by the nucleation temperature and the rate of crystal growth, rather than the duration of the treatment. Thus, by initiating nucleation at ca. 75°C and continuing crystallization at the same temperature, an average crystal particle size of approximately 50 µm is obtained.

Example 3

Preparation of a galenic form for oral administration

3. 1 Gelatin-encapsulated preparations I

A suspension comprising 20% by weight CY-A/X-III obtained in

accordance with the procedure of Example I in Gelucire 44/14 is milled at elevated temperature using a colloidal mill until the average CY-A/X-III particle size is on the order of 0.5 - 10 µM. 250 mg portions of the obtained ground suspension are filled into size 2 hard gelatin capsules, each containing 50 mg ciclosporin (as CY-A/X-III) as active ingredient.

3. 2 Gelatin-encapsulated preparation II

CY-A/X-III obtained in accordance with the procedure of Example 1 is milled in a colloid mill until the average CY-A/X-III particle size is on the order of 0.5 to 20 (IM. 50 mg portions are then thoroughly mixed with 200 g of Cremophore EL and filled in size 2 hard gelatin capsules, each containing 50 mg ciclosporin (as CY-A/X-III) as active ingredient.

Example 4

Preparation of galenic forms for topical application

4. 1 gel preparation I

CY-A/X-III obtained in accordance with the procedure of Example 1 is micronized using an air jet mill. Micronized product with a maximum particle size of

60 - 70 |XM and an average particle size from

2 to 10 JIM are combined and mixed in the usual manner with the component shown in the table below to provide an aqueous gel comprising 10% by weight of ciclosporin (as CY-A/X-III) and suitable for topical application, e.g. for the treatment of psoriasis. 4. 2 qel preparation II

a) is prepared as in 4.1 above, c), f) and g) are dissolved in b) with heating and the resulting mixture is cooled

to room temperature. This solution is dispersed in water h). d) is added and the whole is homogenized to achieve dispersion. a) add and the mixture is homogenized again, e) add to neutrality, with stirring, and the resulting gel is filled into tubes for topical application, e.g. for the treatment of psoriasis.

4. 3 Ointment preparation

a) is prepared as in 4.1 above, b), c) and d) are fused together and stirred and a) added at approximately 30°C, the particles being distributed using a homogenizer. The obtained ointment is cooled and filled into tubes for topical application, e.g. for the treatment of psoriasis.

a) is produced as in 4.1 above, combined with components b) - h) analogously to 4.3 above and

filled in tubes for topical application, e.g. for the treatment of psoriasis.

4. 5 Ointment preparation

A suspension comprising 20% by weight CY-A/X-III (obtained in accordance with the procedure of Example 1) in polyethylene glycol 300 is wet milled until the average CY-A/X-III particle size is on the order of 2 to 10 µm . 10 g of the ground product obtained is mixed in the usual manner with 50 g of polyethylene glycol 300 and 40 g of molten polyethylene glycol 1500 to give a flowable paste comprising 10% by weight of ciclosporin (as CY-A/X-III) and which is suitable for topical application , e.g. for the treatment of psoriasis.

4. 6 Gel preparation

a) prepared as in 4.1 above, b), c) and d) mixed, and f) added and mixed, a) then suspended in

the mixture obtained by homogenization, e) is then added slowly while stirring until gel formation is complete. The gel is then filled into tubes for topical application, e.g. for the treatment of psoriasis.

4. 7 Gel preparation

a) is prepared as in 4.1 above, c), d) and e) are dissolved in b) with heating. The hot mixture is dispersed

i h) by homogenization and cooled to room temperature, a) and

f) is then added and the entire mixture is homogenised, g) is then added and the gel formation is completed. The obtained gel

filled in tubes for topical application, e.g. for the treatment of psoriasis.

4. 8 Cream preparation (water-in-oil)

a) prepared as in 4.1 above and dispersed with b) in e) during heating and homogenization, c) and d) melted

together, mix and the mixture is added to the previous one

obtained dispersion with intensive homogenization, and the whole mixture is cooled to give a fatty white cream. This is filled into tubes for topical application, e.g. for the treatment of psoriasis.

gel/cream preparations

Examples 4.1 - 4.8 above are repeated but with direct use of the crystalline product from example 2 as CICLOSPORIN component.

Example 5

5. 1 Injectable form, e.g., for parenteral administration A CY-A/X-III suspension is prepared under sterile conditions, using standard techniques and using the following ingredients.

The release properties of the active ingredient in the obtained preparation are dependent on the average particle size of component a). Thus, where activity over a longer period of time is required, the CY-A/X-III product with a large average particle size (eg 100 - 200 µM, as obtained directly via the procedure in Example 1) will be selected. Where activity over a relatively shorter period of time is required, a CY-A/X-III product with a relatively smaller average particle size (e.g., obtained by micronizing or grinding the product in Example 1, e.g., into a particulate size from 0.5 to 10 jiM) is selected. The obtained suspension is filled in injection ampoules for parenteral administration or for intralesional injection for the treatment of psoriasis, e.g. to provide a depository effect.

In a particularly preferred embodiment, the example is carried out using CY-A/X-III, prepared in accordance with example 2 above, with a particle size of from 5 to 15 (IM and prepared under sterile conditions.

5.2 Injectable forms, e.g. for intra-articular injection Injectable forms suitable for intra-articular administration are prepared using the following ingredients:

The preparation is prepared in accordance with standard techniques under sterile conditions. Component (a) comprises non-pulverized microcrystals prepared under sterile conditions in accordance with the procedure in Example 2 and with a particle size of from 3 to 15 µM. The preparations obtained are filled, again under sterile conditions, into ampoules for injection, e.g. for intra-articular injection for the treatment of rheumatoid arthritis.

The beneficial properties of the preparations can be demonstrated by means of animal experiments as well as in the clinic. Thus, a particular applicability of the preparations for dermal application, e.g. for the treatment of psoriasis or dermatosis, is shown in the following test model:

ALLERGIC CONTACT DERMATITIS IN GUINEA PIGS (DTH TEST)

Guinea pigs (Hartley, male, 400 - 500 g) are sensitized by applying 50 µl of 2% dinitrofluorobenzene (DNFB) in acetone/olive oil (1:1) to the inside of the right ear. 7 days later, the animals are treated again with 20 µl 0.5% DNFB in acetone/olive oil (4:1) applied to marked areas on the shaved left and right sides. This second exposure induces an allergic inflammation, leading to redness and cellular infiltration (thickening) of the skin. The test preparation in an amount from 200 to 250 mg is applied with a spatula to the DNF-treated area on the right side. The left side is simultaneously treated with placebo as a control. Application of the test preparation/placebo is carried out five times at intervals of 20 minutes, 8 hours, 24 hours, 32 hours and 48 hours after said exposure. The skin thickness at the application site is determined before each application, and again 8 hours after the last application, by pinching the skin into a fold and measuring its thickness. The degree of redness or inflammation is estimated visually on a scale of 0 to 4. The effectiveness of the test preparation in preventing the inflammatory response is determined by comparison with results measured for the placebo-treated sides.

Results obtained in the above test model using preparations in accordance with Examples 4.1 and 4.5 above, but comprising 0.1% by weight ciclosporin as CY-A/X-III, in comparison with placebo and with topically applied 0.1% ciclosporin in normal oral solution form, is shown in the attached fig. I, II and III. [A 0.1% CY-A/X-III concentration is used in this test model for its extreme sensitivity. Test preparations are prepared completely analogously to examples 4.1 and 4.5, but with reduction of the ciclosporin component to 0.1 g and compensation by adding 9.9 g of water in example 4.1 and polyethylene glycol component example 4.5.] In each figure, measured skin thickness is plotted against time. In fig. I are results using 0.1% by weight of regular ciclosporin, applied in the form of the usual, commercial, oral, drinkable solution form compared to placebo. A slight but insignificant reduction in response compared to placebo is seen for the oral solution. In fig. II and III are results using 0.1% by weight ciclosporin-containing preparations in accordance with examples 4.1 and 4.5, respectively, compared to placebo. In both cases, a significant reduction in skin thickening is achieved compared to placebo after the first application and this reduction continues throughout the treatment until the trial is finished.

The applicability of the preparations can also be demonstrated in clinical trials such as carried out as follows: CLINICAL TRIAL: TREATMENT OF PSORIASIS BY TOPICAL (DERMAL) APPLICATION

The experiment is carried out in a random, placebo-controlled design with a double blind test. Each patient receives active preparation and placebo (solution without active ingredient) on two symmetrical lesions at the same time. The distribution of active preparation and placebo preparation on the left and right lesions is random.

Selected individuals are taken from groups of from 12-24 patients, aged 18 - 70 years which include post-menopausal men and women, who have been surgically sterilized or who use oral contraceptives and have a negative pregnancy test.

The criteria for participation include: a) written consent and b) clinically defined, stable, chronic plaque psoriasis with at least two moderate to severe bilateral symmetrical lesions, each with an area of 4 - 25 cm<2> and a score > 5 on a scale from 0-9 in terms of the severity of the lesions. This score is a combination of individual scores from 0 (least severe) to 3 (most severe) for each of the three parameters: erythema, desquamation and skin thickness. Lesions on both sides must be comparable in the same patient (ie, they must not differ by more than 20% in points in terms of severity) and in extent.

The following exclusion criteria are used:

a) Any medical condition which, in the opinion of the researcher, would compromise the study. b) Systemic psoriasis therapy (PUVA, methotrexate, steroids, retinoids, ciclosporin) within the last two months. c) Specific topical treatment - excluding neutral substances (e.g. white vaseline) or preparations containing

salicylic acid (not more than 10%) - on lesions selected for examination within the last two weeks before entering the examination. However, these emollients can be used on lesions for which they are not selected

the survey.

d) Patients whose psoriasis appears to improve spontaneously, which flares up without treatment or regresses on discontinuation

of therapy.

e) Known hypersensitivity to any of the components of the test medication. f) Patients who are uncooperative and unlikely to follow medical instructions exactly, patients who

are unwilling or unable to attend

regular visits.

g) Treatment with a medicinal product to be examined one month before being included in the examination. h) Serious co-occurring disease, especially renal dysfunction with serum creatinine above 130 µmol/l.

i) Hypertension (diastolic blood pressure > 95 mm Hg after 5

minutes in a sitting position). Patients with drug-controlled hypertension are included in the survey.

j) Unmalignant cancer or with previously malignant cancer,

extensive skin cancer,

k) Acute bacterial, viral or fungal-associated

skin lesions.

1) Pregnancy, breastfeeding.

m) Patients with a history including alcoholism,

drug abuse, psychosis or emotional or

intellectual problems.

n) Patients who require continuous and simultaneous therapy with other immunosuppressive agents, chemotherapeutic

agents, compounds with a neurotoxic potential (eg aminoglycosides) and drugs known to affect the pharmacokinetics of ciclosporin (ketoconazole, erythromycin, phenytoin, phenobarbitone,

rifampicin, INH, carbamazepine, sodium valproate).

o) Patients with abnormal laboratory values; i.e. > 15%

beyond the normal limits of the testing laboratory.

p) Patients with clinically significant findings on physical examination

examination.

The experimental preparations used are in accordance with any of the examples 4.1 - 4.9 described above, e.g. preparation B (1%) in example 4.2 or preparation B (10%) in example 4.3.

Each patient receives two coded tubes of 20 g with a two-field label. The label is blue for the substance to be applied on the right side and red for the left side. Furthermore, the side to be treated, left or right, the code for the examination, the researcher's name, the patient's number and the date of the start of the treatment are written in each field on the label. The field that can be torn off the label is inserted into the report folder.

The test drug is applied twice daily, morning and evening, the amount applied depending on the size of the lesion, to give a daily ciclosporin dose of from 0.2 to 2.0 mg ciclosporin/cm<2>, e.g. about 0.5 mg ciclosporin/cm<2> in a test series. Placebo is applied in an equivalent amount. Neither test preparation nor placebo is applied to a body surface of more than 25 cm<2> and no more than 1 g of ciclosporin is applied per week. At each clinic visit, the medicine/placebo tubes were weighed and the topically applied daily dose determined. Patients can bathe shortly before applying the medication. Disposable gloves are put on and changed when applying the experimental preparation and placebo, respectively, to avoid cross-contamination. Allow at least three hours to elapse between application and washing of the treated areas. The first application is monitored and the patient is also given an instruction sheet. The duration of the treatment is four weeks. If the lesions completely disappear earlier, the treatment is continued until week four. The patient is examined before the experiment starts (week -1), at the beginning of the experiment (week 0) and after 1, 2, 3 and 4 weeks.

Dermatological assessment (local psoriasis severity index or LPSI) is assessed by the same researcher, who is not involved in the medical treatment, using the following scale from 0 to 3.

The total LPSI range = value of erythema plus scaling plus thickness = 0 - 9 is evaluated. To be included in the survey, at least two symptoms must be moderate to severe for both selected lesions (LPSI > 5).

At incorporation and at each clinic visit, the area of treated lesions is evaluated (insertion size: 4-25 cm<2> for each plaque). Area refers to the presence of one or more of the three criteria, erythema, scaling, thickening. Pigmentation in the lesion area is not taken into account when estimating it.

The plaque is recorded by photographs before (week - 1) and after the treatment (week 4). Three photographs were taken: one showing both lesions together and one for each lesion.

Itching is evaluated from an interview with the patient using the following scale:

0 = none

1 = minimal: occasionally feels like scratching

2 = moderate: has the urge to itch often

3 = severe: feels like scratching very often, can disturb sleep.

At the end of the study, a global assessment is performed with regard to the response to therapy in accordance with the following criteria:

removed = 100% decline

marked improvement = at least 66% reduction of LPSI,

and/or at least 66% reduction of the 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 the plaque area

worse: worsening of the disease.

Therapeutic progress is defined as any lesion that is "removed" or that shows "marked improvement".

Doctor and patient also indicate their preference regarding medical treatment on the left or right side in accordance with the following criteria:

left side clearly better

left side slightly better

no detectable difference

right side a little better

right side clearly better

To test efficacy at the end of treatment, subject LPSI scores and lesion areas were compared using the WILCOXON Sign Rank test for paired treatments between ciclosporin-treated sides and placebo-treated sides for change in scores from baseline to 4 weeks of treatment.

All further comparisons (at other time points and for remaining efficacy parameters), including the p-values associated with them, are taken as descriptive of progress.

In the above experiments, the subjects show marked improvement in psoriatic lesions, e.g. LPSI scores for erythema, scaling and skin thickness, on the sides to which the test preparation including ciclosporin was applied, compared to the side to which the placebo was applied. Individual improvement is also measured, along with improvement in measured values of pruritus on ciclosporin-treated sides compared to placebo-treated sides.

Positive results are also obtained in trials with the same design, intended for individuals who exhibit contact dermatitis as diagnosed via 3 or more of the following criteria:

- Pruritus

- Typical morphology and distribution:

Flexural lichenification or linearity in adults

Facial and extended involvement in younger and older children

- Chronic or chronic relapsing dermatitis

- Personal or family history of atopy (asthma, allergic rhinitis, atopic dermatitis);

plus 3 or more subordinate moves comprising:

Xerosis, ichthyosis/palmar hyperlinearity/keratosis pilaris, immediate (type I) skin test reactivity, elevated serum IgE, early age at first attack (< 5 years), tendency towards cutaneous infections (especially Staphylococcus aureus and Herpes simplex)/- weakened cell-mediated immunity, tendency towards non-specific hand or foot dermatitis, bumpy eczema, cheilitis, recurrent conjunctivitis, Dennie-Morgan infraorbital fold, keratoconus, anterior subcapsular cataract, orbital darkening, facial pallor/facial erythema, pityriasis alba, anterior neck folds, itching during sweating, intolerance to wool and lipid solvents, perifollicular accentuation, food intolerance, "course" influenced by environment/- emotional factors and white dermographism/long-term whiteness. The trial period is three weeks, with evaluation on a scale from 0 to 3 for the following parameters: pruritus, erythema, exudation, skinlessness and lichenification.

Topical preparations as described above in examples 4.1 - 4.9 containing 1 - 10% ciclosporin, e.g. The 5.0% preparation in accordance with Example 4.7 has also been found to be effective in the treatment of allergic contact dermatitis as well as alopecia.

To support the identification of ciclosporin in orthorhombic crystal form, particularly CY-A/X-III, X-ray diffraction data for CY-A/X-III and for the two forms CY-A/X-I and CY-A/X-II have been provided in the following tables I - III. Attached fig. IV shows the X-ray diffraction diagram from which the data in Tables I to III were obtained. The diffraction pattern was obtained using a Guinier-DeWolff II camera, using CuKα radiation, X = 1.542 Å.

Claims (2)

1. Process for the preparation of ciclosporin in orthorhombic (P212121) crystal form with a crystal lattice of dimensions a = 12.7 Å, b = 15.7 Å, c = 36.3 Å, volume per asymmetric unit = 1804 Å<3>, characterized in that ciclosporin is dissolved in a solvent medium comprising a polymer ether with a high molecular weight, at a temperature of more than 40°C, ciclosporin is crystallized from said solvent medium which carries ciclosporin in orthorhombic (P212121) crystal form as indicated above and the thus obtained crystal form is recovered. 2. Method as stated in claim 1, characterized in that the solvent medium comprises a polyethylene or polypropylene glycol.