PL155712B1 - Method of obtaining novel crystalline form of cyclosporin - Google Patents

Description

REPUBLIC POLAND PATENT DESCRIPTION 155 712 Additional patent to patent No.-- Int. Cl. ⁵ C07K 7/64 Reported: 88 12 20 / p. 276565 / Priority: 87 12 21 United States tniauiu Amephics Η ^Η “ ^a OFFICE PATENT Application announced: 89 08 21 RP Patent description published: 1992 09 30

Inventor

The holder of the patent: SANDOZ A.G.,

Basel / Switzerland /

METHOD OF CREATING CYCLOSPORINS

The subject of invention is a method of producing cyclosporin in a new crystalline form.

Cyclosporin, also known as cyclosporin A, is a known fungal metabolism product with valuable pharmaceutical utility. Cyclosporin is a cyclic N-methylated undecapeptide of formula where -MeBmt- is N-methyl-Z-iZAA-but-iZ-e n-1-yl-4-methyl- / L / treonyl.

Cyclosporin and methods for its preparation are described, for example, in the US Pat. US Anaryki nr 4 117 118. ^^ the main field of clinical trials of cyclosporine concerned its oynorzyntszis as an iI] αtenzruuPrsyJzegr agent, in particular in connection with its use in the treatment of transplant recipients of various organs, e.g. heart, lung, heart and lung transplants, and liver , kidney, pancreas, bone marrow, skin, and cornea, in particular the transitions of allogeneic organs. In this field of application of cyclospoin, success has been achieved so that it enjoys a good reputation and is now widely used in medicine and available under the registered trade name SAliDIMUN or SANDIMMJOT, e.g. in the form of an infusion concentrate, an oral solution and a gelatin encapsulated solution, the latter two variations are disclosed and claimed, e.g., in US Pat. US Aimn / ki No. 4 368 307.

At the same time, other studies were carried out on the usefulness of cyclosporine in the treatment of various autoimmune diseases and inflammations, in particular inflammations with an etiology involving the autotrama component, such as rheumatism / e.g. rheumatoid arthritis, chronic rheumatism and rheumatic rheumatism / and rheumatic diseases, of which reports and results from in vitro, mooel animals and clinical trials are widespread in the literature. Specific α-neurological diseases for which treatment with cyclosporine has been proposed or deserved include: autαemm - norogic haematological disorders / including, for example, haemolytic anemia, aplastic atrophy, anemia associated with ciclosporin155 712

155 712 with 2 red blood cells and primary thrombocytopenia /, sine lupus organ o. »Y, zsdsler.ie cartilage, hardEina, Wegee granuloma.ei-g, dermatomyositis, chronic active hepatitis, myasthenia gravis, psoriasis, Steven's syndrome- Johnson, idiopathic celiac disease, autoimmune inflammatory bowel diseases (including e.g. ulcerative colitis and Crohn's disease), pemphigus, exophthalmia, Graves' disease, sarcoidosis, multiple sclerosis, primary myrhosis, juvenile diabetes / type 1 diabetes /, Behcet's disease, jagodians / anterior and posterior /, hay ketosis, keratitis, interstitial pulmonary fibrosis, psoriatic arthritis, and glomerulonephritis / with or without nephrotic syndrome, e.g. involving idiopathic renal syndrome or minimal renal lesions /.

Further areas of research included the potential usefulness of cyclosporine as an anti-parasitic agent, in particular as an anti-protozoan agent with suggested possible use including the treatment of cold, coccidiomycosis and schistosomy, as well as its use to reduce the resistance of tumors to chemotherapeutic treatments, e.g. cytostatic, and to stimulate hair growth. , e.g. in the treatment of alopecia. In the solid state, cyclosporine exists in both amorphous and crystalline forms.

In the crystalline state, the tetragon and orthorhombic types are known.

The tetragonal variant /P4.,/, also referred to as Cy-// XI for convenience in this specification, is disclosed along with methods for its preparation, e.g., in the Austrainian Patent Specification No. 353,961. Cy-Α / Χ-Ι network paraffiters are as follows: a = b = 1.38 nm, c = 4.12 nm, the volume of the unit ashmtΓłizeβj = 197.4 nm; melting point about 14O-15O ° C; water content - about 2 molecules per molecule of cyclosporine.

The first orthorhombic form / P2,2,2, / is obtained by recrystallization from diisopropyl ether as described in R. Wenger's article Total syn ^ eBie change in moecular structure - biological effect: Ciclosporin as examle in Sandorema 19i8 / III. This particular form is the solvate of the kernel (diisopyl ether) about 1: 2 with the parms of the lattice a = 1.25 nm, b = 2.29 nm, o = 2.84 nm, the volume of the triclinic unit 202.7 ^t of temperature pressure ^p ^ a ^ ° about Ί50 Romtjowe iłklosptryny solvate crystal obtained by recrystallization of supersaturated solutions of cyclosporin in a suitable solvent for longer periods of time. Thus, the orthorhombic solvate form of iłkltspoΓyner / ter diiEopropytowł obtained by Werner is obtained by adding a concentrated cyclosporin solution in mtylene chloride / in which the cyclosporin dissolves relatively well / into the ether for disopropytium / in which the cyclosporin is relatively less soluble / methylene chloride. The residue in the diioσptopytivin ether is left aside to effect crystallization for about 7 days at ambient temperature. The orthorhombic crystalline forms constituting such cyclosporin solvates are also referred to herein for convenience as Cy-A / XH.

By the method of the invention, on the other hand, another orthorhombic form (P2.2.CH2 ^ //) is prepared, characterized in that it contains no or essentially no outside of any soluble component, i.e. it is essentially or exclusively free cyclosporin.

The dimensions of the crystalline lattice of the orthorhombic form of iycltspores are as follows: a = 1.27 nm, b = 1.57 nm, c = 3.63 nm, volume ^of the triclinic unit = 180.4 nm ^{P. The τ} βιηι »βΓβ melting temperature ranges from about 8 ° to 190, and even up to about 195 ° C, depending on the degree of purity and source, as demonstrated in Examples I and II below, which describe specific methods for producing this form. A cyclosporin in the form of neesoiwrtoΛrnłi orthorhombic crystals is also referred to herein for convenience as Ο / Α / Χ-ΙΣΙ. Further characterization data for CY-Α / Χ-ΙΙΙ (X-ray powder diffraction patterns) are given in tetUcy 3 and Fig. 4 below, all with corresponding data for CY-Α / Χ-and CY - // - I.

Notwithstanding the very significant success and reputation achieved by the closptrye, and notwithstanding its significant role in Mtycynite, particularly in the field of organ transplantation, it has become a fact that its practical application is still not ideal or problem-free. Difficulties arose in particular in getting them ready

Conveniently or fully sedative formulations, or most suitable for the treatment of specific diseases or conditions for which cyclosporine has been proven to be useful as a candidate drug.

These difficulties are in part attributed to specific properties of cyclosporin, e.g., its relatively limited solubility or / as has been proven to be compatible with available pharmaceutically acceptable carriers. Such difficulties are related to the peculiarity of certain disease states for which cyclosporine is a potential emergency drug, and to the undesirable side effects that cyclosporin produces when administered orally. For example, cyclosporine has been found to be effective in the treatment of psoriasis when administered orally. Psoriasis is a widespread water disease that suffers considerable suffering in people suffering from psoriasis, for which the generally accepted, effective method of treatment has not been summarized so far. Patients with psoriasis obviously constitute a group of people who need help. While the available data might suggest that cyclosporine may help to meet this need, it is unlikely that oral cyclosporin therapy is considered to be a conventional treatment for psoriasis. Similar considerations apply to the treatment of r.p. eye diseases such as uveitis and conjunctivitis.

Various systems for the administration of cyclosporin have been proposed, including ready-made forms for topical or epidermal and ocular use, e.g. topical application to the surface of the eye, which could be useful in the treatment of psoriasis, other dermatological diseases such as atopic dermatitis and alopecia, or in the treatment of uveitis. So far, however, no significant advantages have been achieved with the use of such systems, nor have it been proven that they are entirely satisfactory.

Thus, there are various reports that the attempts to treat psoriasis patients by applying a known oral solution directly to the surface at the site of psoriatic lesions have ended with minimal success, and in the case of people suffering from atopic dermatitis, this success was largely limited. Although topical application to the eye has been found to be very effective in preventing e.g. corneal graft rejection or in treating diseases of the anterior part of the eye, so far no topical agent has been found that can treat diseases or conditions of the deeper parts quickly, safely or effectively. of the eye, e.g., for the treatment of uveitis, e.g., choroiditis.

Significant difficulties have also occurred in the development of other forms of medicaments containing cyclosporin, e.g., intra-injection medicaments for the topical treatment of rheumatic diseases, e.g., as described above, or for topical injection for the treatment of, e.g., significant psoriasis outbreaks, or other delivery systems. Justnesium, e.g., characterized by a enhanced bioavailability or drug release rate.

Previous attempts to manufacture cyclosporin drugs have used the compound in solution or, less commonly, in amorphous or Cy-A / X-I form, the latter being the identical crystal form for drug registration purposes. However, despite the very intensive search for new or improved variants of the drug, the oral solution and the concentrate for injection are the only commonly used forms of administration of cyclosporine to date.

It has surprisingly been found that new crystalline orthorhombic forms of cyclosporine drugs can be produced by the process of the present invention, which are particularly useful and advantageous for the preparation of various and different types of finished cyclosporin drug forms.

The method according to the invention consists in dissolving the cyclosporin in a medium of a postimetic solvent with a molecular weight of at least 200, containing a thinner orthorhombic rhombic crystal, preferably in a polymeric ether with a molecular weight of at least 200, at a temperature of at least 40 ° C, then the environment is cooled and the crystalline cyclocompatible with the above-mentioned characteristics is recovered.

155 712

In particular, it has been found that such crystalline modifications can be used to produce finished locks containing cyclosporin in a stable, finely divided protein and / or exhibiting improved stability or drug release properties. This discovery opened the way to the production of new and improved cyclosporin-containing drugs, e.g. characterized by a prolonged or delayed release of cyclosporin and / or adapted or suitable for the treatment of diseases or conditions in which treatment with existing agents containing cyclosporin did not give results or in which simple cyclosporine therapy is inherently useful. In particular, it has been found that by the use of such crystalline varieties, especially in the above-mentioned finely divided form, it is possible to produce ready-made medicaments suitable for topical or skin application or topical application to the eye, for the treatment of skin or eye diseases or conditions, e.g. as mentioned above, particularly in the treatment of sclerosis, dermatitis, contact dermatitis, eye symptoms of Behchet's disease, uveitis, and keratoconjunctivitis. The use of such crystalline variants also makes it possible to prepare injectable cyclosporine drugs characterized by a sustained or delayed release and therefore particularly useful, e.g., in the treatment of diseases or conditions for which the increased frequency of administration may make use difficult, e.g. in the treatment of rheumatism or other similar diseases. of joints by intra-articular injection or in the treatment of psoriasis or other similar skin diseases by local injection.

It has been found that for such purposes, orthorhombic crystal modifications, especially w - Α / Χ-, are inherently more useful than other available varieties, in particular amorphous cyclosporin or CY-Α / Χ-Ι, e.g. with respect to physicochemical properties , and more particularly, the recognition characteristics, e.g., the elimination of cyclosporin from the crystal lattice, and suitability for particular drug preparation techniques. CY-Α / Χ-ΙΙΙ produced by the process of the invention is a new and preferred orthorhombic crystal form for the preparation of finished drugs and / or for use in medicine. Accordingly, the invention relates to the balancing of:

A CY - / X-III, i.e., ossilathic orthorhombic crystalline form, and in particular

A ^{1 C} Y ^{- /} X-III parameters sieoi rystα ^{k l} ic and other PHARMACOLOGICAL ^COG ciwośoiao fizyozn ^{h s, h} principle such as defined above, and / or / CY-Α / Χ-ΙΙΙ of the X substantially pro8zżowym as shown below in Table 3 and Inb is shown in Fig. 4.

According to the invention, the method for producing CY - / X-II consists in the crystallization of cyclosporin from an environment containing CY - / X-III oraE in the separation of the thus obtained CY-Α / Χ-.

The equivalents of the media containing H ^^ - / Χ - ΙΙ and used in the above process include, in particular, those that are iyclispiΓin solvents, containing solvent components with a higher mass and particle mass, e.g. with a molecular weight greater than 200. Centers containing higher ethers are particularly suitable. molecular weight, especially pilmlerisiβ ethers, for example polyal ethers such as polyhydric and polypropylene glycols. It has been found that suitable pilmilic ethers include polyhydric and polypropylene glycols, e.g. with a molecular weight of more than 200, glycol esters, e.g. see Fiedler, ^ ^ ϋ der Hilfstoffe, 2nd Edition, Vol. 1, pages 257 and 258), piliocidium esters of sorbitol, e.g. available under the trade name Tween (see Fiedler, Vol. 2, pp. 972-975), polyoxyethylene esters such as polyoxyethylene steayinate ester, e.g. available under the trade name Myyj (see Fiedler, Vol. 2, page 636) and the transesterification products of natural polyalkene polyol ether triterates including corn oil transasterification products, nut oil, almond oil, orzaiha, olive oil, palm oil and ioh mixtures with alcoholic glycols, especially azoza z

Polyethylene glycols with a molecular weight of 200 to 800, e.g., available under the trade name Labrafil (see Fiedler, atrona 539,). Media containing polyethylene glycols with a molecular weight of about 200-600, especially about 300-400 are particularly preferred.

It is clear that the CY-Α / Χ-ΙΙΙ-containing medium may also contain one or more of the other components except those mentioned above, e.g., lower molecular grade alcohols and / or water, as agents that contribute to the solvent characteristics and / or crystallization. A particularly suitable medium for containing CY-1 / X-III has been found to be about 0.5 volume volumes of ethanol, approximately 8.0 volumes polyethylene glycol 300 and approximately 1.0 volumes water.

The process according to the invention consists in the first step of dissolving the cyclnsprin, e.g. in amorphous or tetragonal form, in a suitable medium containing CY-Α / Σ-ΙΙΙ at an elevated temperature. Temperrtura dissolution will of course vary depending on the particular medium chosen, but will suck zazwyonaj ^for from 4 hr) ° C in szczegtilności higher than 50 ° C. * ^{P yp} y ^d to the rod of the ^{COG k} glycol based eolietyleniwym the dissolution temperature is generally higher than 70 ° C, eg. About 75-13 °° C. UEys an ^{angle y} to the solution będnln ^g intented contained ^p thence 5 <preferably more than 10 <eg. To about 40-60S, preferably to not more than 20M ^ oh nfklnspnryny by weight, relative to the total weight of the solution. The crystallization of the cyclosporin is then carried out under cooling, preferably over a relatively longer period of time, e.g. in the order of 10-35 minutes or more, depending on the desired size of the growing crystal.

If necessary, prnoeay nucleation can be initiated, e.g. by ultrasound or seed treatment. It has been found that when the above-described ethanol / enrichedlene glycol / indium system is used as the crystal-containing medium, nuclease can be readily initiated at a temperature of about 60-90 ° C, e.g., about 65-85 ° C, using sonication at a frequency of about 20 ° C. 000 Hz. As shown in Example 1! below, by continuing the sonic treatment throughout the entire period of crystallization, it is possible to obtain micro-crystals with a very small and relatively uniform particle size.

The rate of crystal growth and the size of the crystals obtained will, of course, be dependent on the particular medium selected and on the conditions under which the crystallization is carried out, e.g. whether an acoustic treatment is applied and the frequency of these sounds. When used in the preparation of pharmaceutical drugs as described below, preferably ΟΥ - / Χ-should be used in a finely divided form, e.g. in the form of particles with an average size of less than 200 µm, e.g. 0.5-200 iodine. If relatively larger crystals are initially obtained, then specific formulations containing the desired particle size can, of course, be obtained using conventional comminution methods, e.g., breaking, milling, pulverizing, micronizing, or other methods for reducing particle size. Although the specific preparations obtained in this way may be fully suitable for some applications, such as the place of preparation of ml · jβniwtm agents for application to the skin, the use of breaking or grinding or other suitable techniques in some cases not will be beneficial. The reason for this is that the products obtained by such methods typically have a relatively large particle size distribution and have a relatively coarse and grating texture due to crystal breakage. In addition, disintegration usually results in the destruction of the non-physical characteristics of the natural surface of the crystal, and moreover, it is in any case very difficult or expensive to carry out this operation while maintaining the necessary athenable conditions, e.g. in the manufacture of ready-made medicaments for intra-articular administration. For these reasons, it is usually advantageous to carry out the process according to the invention so as to obtain crystals, e.g. microcrystals, with sizes in the desired particle size range. For example, in the case of using for the preparation of e.g. agents for horizontal application or other horizontal application agents, it will be advantageous to use CY-Α / Υ-ΙΙΙ particle preparations in the form of non-fragmented micro-crystals (i.e. CY-Α / Χ-ΙΙΙ microcrystals in the state of r ^ o ^ dzmni ^ m or non-fine, i.e. those that have not been subjected to breaking, milling, micronization or other procedure leading to a variable particle size reduction /.

155 712

In order to obtain CY-Α / Χ-ΙΙΙ microcrystals that can be used directly in a non-fragmented state in the above-mentioned manner, the process according to the invention is carried out under agitation, e.g. at a speed of 200-300 rpm, increased to e.g. 700 rpm / one minute during crystallization, and in particular uses ultrasound with a frequency in the range of 10,000 - 30,000 Hz, preferably around 20,000 Hz.

By changing the medium containing CY-Α / Χ-ΙΙΙ and controlling the cooling rate and the degree of agitation during recrystallization according to known techniques and as exemplified below, CX-A / X-HI microcrystals with a variable mean particle size, e.g. as set forth above or as will be described below for use in the specific types of finished drug forms that are prepared.

As indicated above, the crystallization according to the invention is preferably carried out under sterile conditions. Purification of the obtained CY-/ Χ-ΙΙΙ can be carried out in the usual way, e.g. by a priming mixture of polyvinyl glycol and water in a ratio of e.g. 3-4: 1 by weight and rinsing with warm water as described in the examples. The obtained CY - / X-III should preferably be reacted in a form containing no or substantially no cyclosporin in any other form, in particular it should be isolated in pure or substantially pure form. The above mentioned washing step helps to prevent the formation of amorphous cyclosporin.

Accordingly, the invention relates to a process for the production of chloropyrins with the above-described honeys, including:

2 / CY-Α / Χ-ΙΙΙ according to any of the items / - / above containing no or essentially no ccclosporin in any other form, i.e. no or essentially no amorphous cyclosporine,

CY-Α / Χ-Ι or CY-Α / Χ-ΙΙ.

/ θ CY-Α / Χ- ^- ! ¹ in accordance with the ^angle óryrokolwiek ^p un ^angles / - / θ formto above in pure or substantially pure form.

c A / CY-Α / Χ-ΙΙΙ according to any of the items / - / again in a finely divided form, i.e. with a particle size defined above or below, e.g. with reference to ready-made drug forums containing CY-A / X-III.

/ ° CY-A / I-III · according to any ^k points / - / θ above containing j mainly ^{or b} exclusively or essentially exclusively unfinished crystalline material, i.e.

CY-Α / Χ-ΙΙΙ according to /, the particles of which are mainly or exclusively or essentially exclusively non-fragmented microcrystals, and /? CY-A / X-II ^and according to the angle ^yellow ryi ^k turtles ^k dots / - / ^ above the ^p ostaci sterile or zasadiiizo sterile, i.e., in a form suitable for pharmaceutical use, in particular, suitable for administration pozajeltowego, e.g. for administration by parenteral injection, in particular for intra-articular injection.

The novel crystalline form of cyclosporin finds use in pharmaceutical compositions containing the orthorhombic crystalline form of cyclosporin as an active ingredient together with a pharmaceutically acceptable diluent or carrier thereof.

The cyclospiria in pharmaceuticals may be of any suitable orthorhombic crystal form, i.e. as CY - / X-II or CY-Α / Χ-ΙΙΙ. The use of CY-A / X-II, however, is generally less favorable due to its physicochemical properties, making it less suitable both for tolerance and for utility in the preparation of effective pharmaceuticals based on cyclosporin. Most preferably, the pharmaceutical compositions comprise cyclosporin in the form of CY-Α / Χ-ΙΙΙ in accordance with any of items / - / above.

Although cycloaporin is present in solid (crystalline) pharmaceuticals, the agents themselves may be in any other form. The pharmaceuticals may be in solid forms such as capsules, tablets, dragees, powders and granules, in semi-solid forms such as avenges, gels, creams and pastes, and also in liquid forms containing the specified cyclosporine component in finely divided form (e.g. and: loosely or microparticulated / suspended or dispersed in a liquid diluent or pharmaceutical carrier in which the selected crystalline form can be retained.

Pharmaceuticals include both single phase or compartment systems with cyclosporine, e.g. simple creams, gels, venges etc. as mentioned above and simple one compartment capsules or capsules, e.g. hard or soft gelatin capsules and injection ampoules. as well as systems involving several phases or compartments. Thus, also include agents comprising a phase or compartment containing cyclosporin together with one or more phases or compartments not containing cyclosporine, such as shell, carrier, partition or placebo phases, or phases or coatings containing other therapeutic substances or additives. Therefore, these measures include, for example, coated tablets containing a core with cyclosporin in the orthorhombic crystal form of otoozones, e.g. with other coating phases not containing cyclosporin, etc., tablets with a shell with a core or a shell containing cyclosporin in an orthorhombic crystal form and with a shell or core cyclosporin free mixed granules include a first granulate phase containing cyclosporin in orthorhombic crystalline form and a second granule phase free of cyclosporin, cleavable capsules Containing two or more compartments, e.g. rhombowwj and the other does not contain it. Other variations and variants are apparent to those skilled in the art.

The pharmaceutical compositions contain cyclosporin primarily or, preferably, exclusively or substantially exclusively in the orthorhombic crystal form, e.g. as CY-Α / Χ-.

The pharmaceutical compositions preferably contain at least one lab phase compartment comprising cyclosporin in an orthorhombic crystalline form, e.g., CY-Λ / Σ-, in which the crystalline form is evenly distributed or evenly distributed. Such agents include, for example, homogeneous creams, venges, gels and the like, as well as liquid agents in which the cyclosporin content is evenly distributed or can be evenly distributed in an easy manner, e.g. by shaking the hand.

The pharmaceutical compositions may include those suitable for topical application, e.g., for application to the skin or topically to the eye, for parenteral administration, e.g. by infusion or injection, including by subcutaneous or intramuscular injection, and in particular for local injection. or intra-articular administration, as well as intravenous administration means, e.g. suppositories, pessaries and the like, and oral agents. Enteral agents, e.g., oral agents, are generally less preferred, however.

Preferably, the orthorhombic crystalline cyclosporin, e.g., CY-Α / Χ-, is in finely divided form in pharmaceuticals, e.g. The appropriate particle size will of course vary depending upon the particular type of agent, intended use, mode of administration and desired effect. In principle, however, a suitable mean particle size is about 0.5-200 µm. Specific size ranges, for example with regard to oral, topical or injectable agents, will be described in more detail below.

Agents for topical administration, e.g. for application to the skin or topically to the eye, especially in the treatment of psoriasis or atopic dermatitis, or for eye diseases or conditions requiring treatment for imminotrophic therapy, e.g. uveitis, conjunctivitis including conjunctivitis and rhinitis hay fever or corneal transplant.

Gels, creams, venges, etc. for topical application and skin application may be prepared by any of the known and commonly used methods, e.g. by mixing cyclosporin in an orthorhombic crystalline form with a suitable liquid carrier such as an aqueous gel, e.g. a hydrated gel cellulose ether, with the addition of certain surfactants, thickeners and the like, including e.g. aerosil.

The cyclospoyine component is preferably present in finely divided form, e.g. with an average particle size of 0.5-200 µm. The mean particle size is ^! conveniently below

155 712

60-80 µm, preferably less than 40 µm, even more preferably less than 30 µm and most preferably less than 20 µm. The particle size distribution is preferably as narrow as possible. Preferably substantially all of the particles are less than 60-80 µm.

Agents for topical application to the eye, for example, for application to the eye, may include, for example, suitably thickened suspensions or dispersions, for example presented as viscous eye-drop formulations or eye creams or gels. For ophthalmic agents, the average particle size of the crystalline cyclosporin material is preferably less than 25 µm. Even more preferably, substantially all of the particles have a size below 25 µm. The average particle size of such formulations as eye drops is suitably from about 0.1 or preferably from about 0.5 to about 20, preferably to about 10 µm. The content of the hydrophilic / lipophilic components in the vehicle for use in dermal application agents or in vehicle formulations may vary depending on the particular surface to which the agent is applied, e.g. depending on the properties and characteristics of the particular skin area to be treated.

Compositions for topical application, especially to the skin, preferably contain a carrier in which the active ingredient (cycloaposin component) is only slightly soluble. The solubility of the cyclosporin component / e.g. The 0Υ-Α / Χ-ΙΙΙ / in such compositions is suitably less than 104, preferably less than 1.04. Even more preferably the solubility is less than 0.14 and most preferably less than 0.0014.

Suitable carriers include hydrophobic water-miscible carriers as well as hydrophobic carriers. The hydrophilic, water-miscible components of the carrier are preferably components with a relatively low volatility. Examples of the carrier components belonging to this group are liquid polyalkylene glycols, propylene glycol, glycerin and ethylene glycol. Suitable polyalkyl glycols include, in particular, liquid polyethylene glycols, e.g., having an average molecular weight of up to about 600, such as having an average molecular weight of about 250-450, and especially about 300.

Examples of topical agents include swirling agents:

C ¹ a / yklos ^ y ^ ^ in staoi ruble solvated preferably N ^ ^ h ^c r ^{y s, t} s is rhodium, especially 0Υ - // Χ-ΙΙΙ eg. As determined in accordance with the points AA * powyyżj b / hydrofUowy a water-miscible vehicle as defined above; and optionally c / pharmaceutical grade water.

The amount of component (a) is suitably about 0.1-30.04, especially about 0.5 or 1.0 to 204, e.g. about 1.0, 2.0, 5.0 or 10.04 based on total weight of lomooydia. The components / b / and / c / are conveniently used in a ratio of from 1: 0 to 1:20 by weight. More particularly, the ratio of components (b / and / c) is from 1: 0.5 to 1: 5, e.g. from 1: 1 to 1: 3, most preferably from 1: 1.5 to 1: 2.5 by weight.

Agents of this type additionally also contain an air-borne agent.

Relevant components / d / include, but are not limited to:

'd¹ Produ^kvol^ytΓansl8try^fikacj^and natural ^ and tΓig^andieβry^thisin the winters of ro ^ i-nn ^ h^at pioIIoI polyalkylene. Such transesterification products are generally known and can be produced, e.g., in accordance with the general juveniles disclosed in US Pat. US Amyki No. 3 288 824. These include transesterification products of various natural (that is, nile-resistant) vegetable oils, e.g. corn oil, nut oil, almond oil, ground nut oil, olive oil and palmlwlgl oil and their mixtures with glycols polyethylene glycols, especially polyethylene glycols, with an average molecular weight of 200-800.

Preference is given to the products obtained as a result of tΓθesβ8trification of 2 mole parts of natural oil trigicceide mixed with 1 part of plliθdyeen glycol / e.g. with an average molecular weight of 200-8 (0) /. Various forms of the transesterification product of a particular group are known and commercially available under the trade name Labrafil (see Fiedler, lerilren der Hilfstoffe, 2nd edition zliθeelee and extended (1981), Voi. 2,

155 712 page 539 /. The following products are particularly useful as ingredients of cfo: Labrafil 1944 CS, a transesterification product of nut oil and polyethylene glycol, with an acid number of about 2, a reduction number of 145-175 and an iodine number of 60-90, and Labrafil 2130 CS, a glyceride transesterification product C 1-8 C with polyethylene glycol, a melting point of about 35 ° C to 40 ° C, an acid number less than 2, a haze number of about 185-200 and an iodine number less than 3.

d The products of reaction of natural or hydrogenated castor oil with ethylene oxide. Such products can be prepared in a manner known per se, e.g. by reacting natural or hydrogenated castor oil with ethylene oxide, e.g. in a molar ratio of about 1:35 to about 1:60, with optional removal of the polyethylene glycol component from the product, e.g. with honeys disclosed in German Patent Laid-ins Nos. 1,182,388 and 1,518,819. The various surfactants available under the trade name Cremophor are particularly useful. There are, in particular, Cremophor RH 40 products with a saponification number of about 50-60, an acid number below 1, an iodine number below 1, a water content (determined by the Fischer method) below 2%, η ^ θ about 1.453-1.4578 and on balance hydrophilic / lipophilic / LHB / about 14-16, Cremophor RH60 on the number d ^ laiia about 40-50, an acid number less than 1, an iodine number less than 1, the content of water ^s / fisoher / 4.5-5.5%. B ^p25 oltole ¹ , 453-1 ^, 57 and MB oltole Ί5-17 ^, ero Cremophore BL with a molecular weight / determined by the vapor osmometry method / about 1630, the number of 25 washing (Hania about 65-7 °, l-acidic acid about 2, the number is around ²⁸ ~ ^{32 and} around

1.471. Various surfactants available under the trade name Hikkol, e.g. Nikkol HCO-60, are also suitable for use in this group. The exuded Hikkol HCO-60 agent is a reaction product of hydrated castor oil with an ethylene bladder, with the following properties: acid lipid 0.3> leakage number 47.4, hydroxyl number 42.5, ^pH / solution 5% ^/ 4, ⁶ , ^b Arw of HA and ^p 4 ^0, the temperature ^{t and} a ³ opnien 6, ^O ° C, ^for mrzania 32.4 ° C, the water / Karl Fiscluar / 0.034.

d ³ esters kwasdw łuszcow ^{t s} c ^{h t e} oliok8ye ylerowanym sorMten one half: i.sor ^{b iniany, n} p. of the type known and commercially available under the tradenames Tween and śrmotan / c.f. Fiedler, Vol. in, pages 745- 746 and 972-975, including Tween products

- monolaurinate ^ol eoxyetyle no / 20 / aorbitu

- monoleaIBeeyniαo eoli · ok8ye so much no / PO / sorbitol

- mololte8Γyniao poly-okayetyle no / 20 / sorbitu

- polyoxyethyl triaeearate / PO / sorbitol

- ellyoxyethyl nO / sorbitan monolleate

- plylxyethyl-sorbitan trioleate

- monoOauryniαo polylxyetyle noA / sorbitol

- mololtβaryniao pllilk8yetyle noA / sorbitu and

- pllloxyetyleoo / 5 / sorbitol molooleate.

^d5 Polio ^k s ^y itylelowi ester ^k was <5w fatty example. ^ lioksyety ^ these esters atear ^ of the type known and commercially available under the trade name Myrj / c.f. Fiedler, Vol. 2, page 636 and / poliok8yθtyiolowe esters tłuBcdowych known and commercially available under the trade name Cetiol HB (see Fiedler, Vol. 1, page 22Θ) and polyoxyethylene fatty alcohol ethers, e.g. ,

Vol. 1, 1 & 4-186), e.g. Brij 78 and 96 and Cetomacrogel 1000 (see Fiedler, Vol. 1, 229). cd copolymers EOLIE ⁰ k ^c syβtyiolowl-pllloksypropylenowe eg. of the type known and commercially available under the trade names Plunnic and Bmmalyz / c.f. Fiedler, 2, pages 720-722 /. d ⁵ esters of sorbitan ^b and ^t u acids Huzcc uwymi eg. t ^y j) in a known and Preliminary commercially ^ ¹ the trade name Span, which include eoool8urynian, mololeCmetyniao, mololataΓyniao, triseearynian, monoolθioian trioleate, sorbitan / c.f. Fiedler 2 , pages 850-851 /. d Partial glycerides, e.g. products containing mono and / or triglyceride components, e.g. mono, lithium and triglyceride mixtures, and free glycerol as well as e.g. glycerol monosearates and glycerol monosuricates.

155 712

Examples of such products are those commercially available and known under the trade names Imwitor (see Fiedler, 1, 491), e.g. Imwitor 191 and Imwitor 780 and Softigen (see Fiedler, 2, 833), e.g. Softigen 701.

^d ® ionic środlti ^p owierzchniowo part nne ^{t y t} emulsifying WHAT I ^ b ^ ^k osiarczan s £> for ^you cetostearylosiarcEan sulfate.

If present, ingredients (d) are suitably up to 30%, more preferably up to 15%, based on the total weight of the agent. Most preferably the content of component (d) is about 0.5-10%, e.g. about 0.1%, based on the weight of the agent.

Pharmaceuticals of this type will conveniently also contain an e / thickening agent.

Relevant components / a / are, for example:

e ¹ Żywioe pollaa loakrylanowe ^you like. Known ^you access ^g of i. ^h andlu available when the name ^and the ^{d h} aadl ^of hose Budiapart / c.f. Fiedler, 1, 371-372 /.

e Cellulose derivatives which include ethyl, propyl, methyl, hydroxypropyl methyl and carboxy methyl cellulose.

E ^ ^{p ZY} hoses olivine lowe ^s, n ^p. of ^b ejmuj ^± sun polyvinyl wine ^yl pink and poliwinylop: Lrolideny ^and other rateriały polymer such as gelatin, alginates, pectins, gum tragacanth, gum arabic, xanthan gum, and polyacrylic acids such as those known and available commercially under the trade name Carbopol (see Fiedler, 1, 206-207).

e ^ has ^a trio ^{BRIEFINGS t} ak: e ^f as above. ^b entoni.ti silica aluminum silicate magnezoww.

If components / e / are present, the amount is suitably up to 20%, even more preferably up to 10%, of the total weight of the agent. Most preferably, the amount of ingredients (e) is about 0.5-15%, e.g. about 1.0% - about 3.0%, of the total weight of the agent.

In addition to the above ingredients, the composition also conveniently comprises an antimicrobial agent such as methyl paraben, propyl paraben, benzyl chloride or benzyl alcohol, e.g. in an amount of 0.05-0.5%, and in the case of benzyl alcohol up to 2.0%, e.g. about 0.1% and, in the case of benzyl alcohol, about 1.0%, based on the total weight of the agent.

Alternatively, the carrier is made of hydrophobic materials, e.g., those which satisfy the solubility requirements for the active ingredient, given in ibid. * therefore, further preferred pharmaceuticals, e.g. for topical use, include:

ABOUT

C a) Cyclosporin in the form of orthorhombic crystals, preferably nitro-α7 orthorhombic crystals, e.g. CY-0 / Σ-określoną as defined in any one of / - / above and f) a hydrophobic support, e.g. as defined above.

The amount of component (a) is conveniently the same as given above for the agents defined in C ¹ . Suitable components / ^f / n are ^p. petroleum derivatives, including mineral oils, fats and jellies such as e.g. petrledum (mineral fat or petroleum jelly), liquid petrolaumm / white mineral oil or liquid paraffin / or any other products known and commercially available under the trade name Vaseline (patmE Fiedler, 2,986,) and lanolin-based nonionic products such as those available under the tradename Anechcl, e.g. C (B (Fiedler, 1,119-120)).

Other suitable ingredients (f) include, for example, natural or saturated vegetable oils and fats, e.g. k-k-s-wβgo nut shell oils such as those known and commercially available under the trade name Miglyol, e.g. Miglyol 812 (see Fiedler, 2,

616 / ογθε artchis oil and seed peanut oil, as well as higher fatty alcohols and R-B-chain fatty alcohols, and fatty esters such as srtyl alcohol, cetyl palmitate and 2-octylde-dtkanol, which is known and commercially available under the tradename Butan-G (see Fiedler, 1,375).

Such agents may also contain one or more of components / d / and / e / as defined above as well as one or more anti-aggressive agents, e.g.

In particular, such compositions may additionally contain one or more components (d), and generally in an amount of up to 30%, even more preferably not more than 20%, and iogoinite in an amount of about 1-15%, by Weight based on the total weight of the agent. .

155 712

Agents based on hydrophobic according to C? also containing the emulsifying component / d / can also be made into creams or emulsions by adding water thereto. The resulting creams or emulsions may be oil-in-water or water-in-oil type. The water content of these products can be, for example, 5-805 based on the total weight of the agent, typically 20-705 µ.

If incorporated with / e / and antimicrobial agents, the quantities may be of the same order as or similar to those in the above amounts of carrying the ^COG set to środkcfo lon c ^y CH \

The amount of the topical agent applied will of course vary depending on the concentration of the agent, the condition of the disease being treated and the effect desired, and also, in the case of application to the skin, on the surface to be coated. For dermal application agents, e.g., in the treatment of psoriasis or dermatitis, the amounts applied will typically be such as to provide a cyclosporin dose of from about 0.1 to about 5 mg / cm, e.g., about 0.5 mg / cm. when used twice a day on the desired site.

In the case of oozing formulations, administration can be such that formulations containing about 0.1-10%, e.g. by weight of cyclosporine, use drops in each eye 1-3 times a day.

Injectable forms, e.g. for subcutaneous, intramuscular or other parenteral injection, in particular for intra-articular injection, e.g. in the treatment of rheumatoid arthritis, or for intrauterine injection, e.g. in the treatment of psoriasis.

Dispersions, suspensions, emulsions and other similar injectable agents can be prepared according to any of the known and commonly used methods, e.g. by dispersing, suspending or otherwise dispersing cycSoaporin as finely divided orthorhombic crystals in a suitable liquid carrier with the addition of suitable evugators, surfactants, stabilizing or flocculating agents and the like. Preferably, such agents have an average particle size of the cyclosporin component below 100 µm, even more preferably below 50 µm, and most preferably below 20 µm. Suitable mean particle size should range from 0.1 or 0.5 up to oxidized particles greater than 20, 50 or 100 µm.

When the injection means are intended, for example, for subcutaneous or intramuscular injection, for example to obtain a reserve effect, the use of larger particles is permissible. Where such means are intended for local injection or, in particular, for intra-articular injection, it will be preferable to use a formulation containing finer particles. For such applications, a suitable particle size is e.g. on the order of from about 0.1 or about 0.5 to about 20 µm, e.g. about 5-15 µm.

In order to ensure particle stability in the injection means, e.g. to avoid the larger crystalline particle size increasing at the expense of smaller particle size, the particles should preferably be as uniform in size as possible. Preferably the difference between the size of the largest and smallest particle should not exceed 50 µm, and even more preferably about 20 µm.

Although the desired particle size can be obtained by using appropriate comminution methods, such as micronization, in order to reduce differences in particle size, eliminate the presence of broken particles, avoid surface modification, e.g. to maintain proper wettability and to more easily meet the necessary As sterility requirements are concerned, it is usually preferred to use non-fragmented microcrystals, i.e., a crystal grown as described above so as to meet the size requirements.

The injectables therefore contain, respectively:

C a) cyclosporin in the form of finely divided orthorhombic crystals, advantageously unsolvated orthorhombic crystals, such as CY-Α / Χ-ΙΙΙ, e.g. in the form defined in accordance with any of the points AA, especially A and A, distributed or giving spread in b / in the injectable medium.

155 712

A suitable carrier is in particular pharmaceutical grade water.

Agents according to <3 may additionally also contain g / surfactant as dispersing or emulsifying agent and / or flocculating agent to aid in spreading or to keep the particles distributed in the carrier phase.

For idpiwiednich surfactants as components / g / tłusczowwych are fatty acid esters of sorbitol or poliok8yetyleniwanym polysorbates, eg. Such as ^{the no} supply piinkcla d ^ ⁱⁿ the yżej such as Twwen 80 l.ub Ροϊ ^ ϋ ^ β ^ 80 / see Fiedler, 2, page 746 /. Suitable flocculating agents / g / include ttylnidylaminotetraacetate and its salts, e.g. sodium EDTA and calcium BDTA, as well as other suitable chelating and peptizing agents such as succinic acid or citric acid.

A stabilizer is also preferably used to maintain a proper grain size distribution. Suitable stabilizing agents include, for example, gelatins or modified gelatins such as the plasma expander known and commercially available under the trade names Gelafundin and Haemoool, as well as cold water soluble gelatins from finely purified hydrolytes and collagen thibles.

Means own preferences ^{g C p} owyżej may contain one Rownica February), more ^{funding does} Ex ^ iwdrobnoustrojowych, for example. As described powyyżj, and means peptydyzujące such as succinic acid or citric acid.

The weight ratio of components (a): / b) in the compositions defined in <3 is suitably from

1:10 to 1: 1000, preferably 1:50 to 1: 200, e.g. about 1: 100.

In order not to unnecessarily compromise the stability of the particles, the surfactant concentrations used as components (g) should preferably be relatively low, e.g. and/. If a flocculating agent is used, its weight ratio to component (a) is and is suitably from 1: 3 to 1: 8, e.g. about 1: 5. If a stabilizing agent is used, the weight ratio of component (a) to stabilizing agent is suitably from 1: 5 to 1:30, more preferably from 1:10 to 1:30, e.g. about 1:20. If a peptidizing agent is used, the weight ratio of peptidizing agent to component (a) is suitably from 1: 1 to 1:10, e.g. about 1:15.

The amount of ciclosporin in a unit dose for injection will, of course, vary depending on the disease being treated, the site of injection and the effect desired, e.g. whether delayed action or relatively rapid release into surrounding tissues are envisaged. A suitable unit dose should be about 10-500 mg, more preferably about 20-100 mg, per cycle.

It is understood that unit dose forms also encompass more complex systems, e.g., dual chamber syringes or syringe barrels, wherein the comminuted cyclosporin along with other suitable additives, e.g., stabilizer and antidibacterial agent, is contained in the first chamber and the carrier in the second chamber. the components of both chambers are depressurized, e.g. by depressing a syringe plunger. Such two-chamber systems are well known in the art and will be understood to fall within the scope of the unit dose form containing the formula according to the invention.

Oral dosage forms, e.g., for the treatment of autoimmune diseases and other conditions mentioned above, in particular for the prevention of transplant rejection.

Although, as noted previously, the oral dosage forms are generally less interesting and therefore less preferred, they may also contain ctclospirin prepared by the method of the invention.

Oral dosage forms can be prepared utilizing any suitable carrier or carrier medium that is commonly known and used. Oral liquid or semi-solid dosage forms may, for example, contain cyclosporin in an orthorhombic crystalline form, e.g., CY-Α / Χ-wraz, together with pharmaceutically acceptable diluents or carriers in which the crystalline phoma is insoluble or substantially insoluble. Such carriers and diluents include, for example, transesterification products

155 712 natural vegetable oil triglycerides with polialkiloanopoliolami described above in / d ¹ / such as ^{p y} rodukt Labrafil, and the reaction product ^{s 1} J.ub natural uwodornlonego tlenkCem castor oil with ethylene, such as those described above in the / d /, especially Uremoohora liquid products, e.g. Grcmophore BL.

The cyclosporin is preferably in a finely divided form, for example having an average particle size of 0.5-200 µm, especially of the order of about 0.5-30 µm, preferably less than 20 µ, eg 0.5-10 µπι.

For convenience of administration, compositions of this type are suitably filled into hard or soft gelatin capsules.

Dosage unit forms for oral administration conveniently contain from about 25 to about 75 or up to 200, e.g. about 50 or 100 μm cycloaporin / unit dose, such drugs are administered e.g. 1-4 times daily to achieve a specific patient concentration. serum cyclosporins / e.g. by radioimmunoassay / of the same or equivalent order as is achieved with conventional lycllβ porinuction therapy, e.g. with the available cycle oral solution.

It is understood that the ingredients listed above or defined as pharmaceutical ingredients should be suitable for the intended pharmaceutical use. Thus, for the ingredients of an orally administrable agent, they should be pharmaceutically acceptable, and for the ingredients of topical agents, they should be acceptable or tolerable, and suitable for topical application, e.g., to the skin or eyes.

Other formulated drugs are also apparent to those skilled in the art for use in equivalent or different methods to those described above.

The following examples illustrate the process of the invention in each of its various aspects

Example I. CY-A / X-III challenge.

a / 100 g of cyclloor ^, y and y in amorphous form are dissolved with stirring in 400 g of polyethylene glycol 300 at 50 ° C. After the dissolution is complete, the rotifers begins to appear CY - / X-III. The solution is left for 24-48 hours at a temperature of 35-4O ° C to complete crystallization and then diluted with 1000 ml of water with intense stirring. The precipitated solid is filtered, washed with water to remove residual polyethylene glycol, and dried to yield CY- / Y-III in essentially pure form; Melting point 195 ° C, particle size - about 100-200zum.

CY- / X-III can be othylated in a manner analogous to that described above if the following materials are used as solvents in place of poHey! Melting solvent obtained

C - A / X-III / ° C / b / olliety glycolCci 300 / Crcmophorc RH 40/1: 1 by volume / c / Cremophorc EL 190 d / Solutol HS 15 188 a / Gelucire 44/14 180 f / Myrj 52 184 g / Twwen 80 186 h / Crcmophor RH 40 190

In each case, particles having an average size on the order of about 50-200 µm were obtained. The determined lattice structure was as stated in the following C · X-ray diffraction patterns were as shown below in Table 3 and Figure 4.

Example II. Producer CY - / X-III. 20 g of cyclosplrin in the 0Υ-Α / Χ-1 form are dissolved in 60 ml of ethanol and then concentrated to 1/6 of its original volume by distillation under reduced pressure at a temperature below 60 ° C. A mixture of 160 ml of polyethylene glycol (molecular weight 300) and 20 ml of water is then slowly added at a temperature of 50-85 ° C while stirring at 300 rpm. An ultra-tube vibrator / mixer operating at 20,000 Hz is installed. Hucleation

155 712 is initiated at 65-75 ° C in 3-5 minutes. The dispersion obtained is cooled at a constant speed for 10-20 minutes to 40 ° C with constant stirring at 700 rpm and with the aid of ultrasound. The obtained thick pasty mixture is filtered, rinsed with a mixture of polyethylene glycol 300 / water 3: 1 by volume, washed with warm water at 30 ° C and dried at 50 ° C under high vacuum, obtaining CY - / X-III in a pure form at temperature mp 192 ° C. Particle moisture - 3-15 µm. The marked hay structure was as stated above. The X-ray diffraction patterns were as shown below in Table 3 and Figure 4.

Experiments have shown that the size of the crystals is influenced by the nuclease temperature and the rate of crystal growth rather than by the duration of the operation. Thus, by initiating nucleation at about 75 ° C and co-crystallizing at the same temperature, crystals having an average size of about 50 µm are obtained.

The beneficial properties of the agents containing cyclosporin according to the invention can be demonstrated in experimental animal experiments and in clinical trials. The particular suitability of agents intended for application to the skin, e.g. in the treatment of psoriasis or dermatitis, can be demonstrated in the following model study. Allergic contact dermatitis in guinea pigs / DTH test /.

mo ^ kie / Hartley, roe deer, 400-500 g / sensitized by introducing 50/1 2%. dieitroflrorobθnEenu / DNFB / in a mixture of acetone / o ^ wa / 1: 1 / on the inner side of the right ear. 7 days later, animals are re-treated with 20 µl 0.5%. DNPB in an acetone / 4: 1 mixture / applied on the marked areas on the shaved left and right side. This second challenge treatment causes allergic inflammation leading to reddening of the cellular infiltration / thickening / skin. The test agent in an amount of 200-250 mg is applied with a spatula to the DNPB-treated area on the right side. The left flank was similarly trained with a placebo on a control cell oh. Test agent and placebo were applied five times at 20 minutes and at 8, 24, 32 and 48 hours after challenge. The thickness of the skin at the application site was measured before each application and after 8 hours. The measurement was made by lifting the skin in the form of a fold and measuring its thickness. The degree of redness and inflammation were also assessed visually on a scale from 0 to 4. The efficacy of the test agent in preventing the inflammatory response was determined by comparison with the results recorded for the placebo sides.

The results obtained in half the mooel test with agents containing 0.1 wt.% Of cyclosporin as CY-Α / Χ-, compared to planbo and topical cyclosporin in the form of the usual 0.1%. solution for oral use is shown in Figures 1, 2 and 3 of the accompanying description. In this mcoel test, the concentration of 0.1% was used. due to its exceptional sensitivity. The bacon products contained 0.1 g of the CłklospoΓynowlgj ingredient.

Each plot shows the measured skin thickness as a function of time. In Fig. 1, the results obtained using 0.1%. / w / ccyclosphate used in the form of a common, commercially available oral solution to be drunk was compared with a placebo. There was a slight but insignificant reduction in response compared to placebo. In Figures 2 and 3, the results obtained with agents containing 0.1% by weight of cyclosporin were compared to placebo. In both cases, a significant reduction in skin thickness is achieved compared to placebo starting from the first application of the agent, throughout the treatment and ending with the trial.

The utility of the agents containing cyclosporin according to the invention can also be demonstrated in clinical trials, e.g.

Clinical Trial: Treatment of psoriasis by topical application to the skin

The assay was performed according to a double-blind, placebo-controlled, averaged design. Each patient was dosed with drug and placebo (vehicle alone) simultaneously on two symmetrical disease lesions. Allocations of active agents and placebo were made statistically for the left and right lesions.

155 712

Selected individuals were divided into groups of 12-24 patients aged 18-70 years, men and women after the menopause period, surgically clean or such,

Which were taking oral contraceptives with a negative pregnancy test.

The criteria for conducting the study included: a / written consent and b / clinically proven, persistent, chronic lobar psoriasis in the form of at least two bilateral, moderate to severe, syrotic foci, each with an area of

4-25 cm and with a severity degree above 5 on a scale from 0 to 9. This score was the sum of individual scores from 0 / least severe / to 3 / most severe / for each of the three parameters: erythema, peeling and skin thickness. The foci on both sides for a given patient must be comparable, that is, they cannot differ by more than 20% in terms of their degree of severity or surface area.

The following exclusion criteria were used: a / health condition which, in the opinion of the investigator, could challenge the results of the study, b / traumatic treatment of psoriasis / PUVA, raehotreiate, steroids, retinoids, cyclosporine / within the last two months.

c / specific local treatment - with the exception of inert substances / e.g. white petroleum jelly / or agents containing salicylic acid / no more than 10% / on fireplaces for research in the last two weeks prior to the commencement of the trial. Such softeners, however, can be used for outbreaks not selected for research.

d / patients who have been shown to have milder psoriasis, erupt without treatment or if it is discontinued.

e / known hypersensitivity to any of the components of the tested drug.

f / poorly cooperating patients who do not follow the doctor's instructions strictly and patients who do not want to or cannot undergo regular visits.

g / treatment with study drug within one month prior to study start, h / acute comorbid disease, especially renal abnormality with serum creatinine above 130 µmoles / liter.

and / hypertension / diastolic blood pressure over 95 mm of mercury and after 5 minutes in a sitting position /. Patients with medication-treated hypertension were included in the study.

j / malignant neoplasms or previous malignant neoplasms, including skin cancer.

k / acute inflammation of the skin of bacterial, viral or fungal origin.

1 / pregnancy, breastfeeding period.

m / patients with a history of alcoholism, drug abuse, psychoses or emotional or intellectual problems.

n / patients who undergo permanent treatment of other diseases with other immunotherapy agents, chemotherapeutic agents, nephrotoxic compounds / e.g. emirtglikoEydów / and drugs known to affect the pharmacokinetics of cyclosporin Αιϊο ^ ιθζοΙ, erythromycin, pheny ^ ina, phenoba ^^^, rffampicbne, INH, ka ^ aze?! ^, sodium salt of 2-propyl valerate acid /.

o / patients with abnormal baseline analytical results ^ exceeding the normal limits of the assay by more than 15%.

p / patients with clinically significant symptoms that make diagnosis difficult.

Measures such as' 1 percent. middle or 10 percent. center.

Each patient received a pair of 20 g coded tubes with a two-piece label.

The label was blue for the substance to be applied on the right side and red for the left side. Each part of the sticker also includes the side to be applied, left or right, the test code, the name of the person conducting the study, patient number and the date of treatment initiation. A detached portion of the label was attached to the test report form.

The test drug was applied twice a day, morning and evening, the amount applied depending on the size of the focus, so as to obtain a daily dose of ąyclo8ptry of 0.2-2.0 m> / <m, e.g. in one series of trials about 0.5 mg ąbklt8poIbTb / cm. Placebo was applied in

155,712 of an equivalent amount. No study drug or placebo was applied to a body surface greater than 25 cm and no more than 1 g of ciclosporin / week was applied. At each clinic visit, the drug and placebo tubes were weighed to determine the daily topical dose. Patients may take a bath shortly before taking the drug.

Disposable gloves are worn each time to apply test and placebo to prevent cross-contamination. At least three hours should elapse between applying the medicine and washing the treated area. The first application is performed under supervision and the patient is given an instruction sheet. Lying down lasts 4 weeks. If the disease heals more quickly, treatment is continued until the fourth week. Patients are examined before trial start (week 1), trial start (week 0) and after weeks 1, 2, 3 and 4.

The dermatological assessment / local psoriasis severity index or LPSI / is performed by the same investigator, who is not a patient's physician, using the following numerical scale from to 3.

Erythema = none = minimal: 1 erythema is poorly detected; very light pink base = moderate: visible pale -Eewone coloration

Flaking = none = minimal: poorly developed> areas of flaking = moderate: clearly defined areas of flaking starting to clump

Skin thickness O = none = minimal: hardly perceptible thickening = uM-sheet: clearly palpable thickenings = silty:

dark / light red color = strong: clearly defined areas of flaking, compacting oh = strong: strong thickening

The total LPSI size is assessed = erythema size plus desquamation plus thickness, ranging from 0 to 9. To initiate the study, at least two signs must be moderate or severe for both selected foci (LPSI greater than 5).

At the beginning of each clinic visit, the area of the treated focus is measured / size

Output 4-25 cm / fire /. Areas are scored against one or more of three criteria including erythema, flaking, and thickness. The pigmeetaaji inside the area of the outbreak shall not be taken into account for the assessment of that area.

Foci are documented by photographing for / week - 1 / and after examination / week 4 /. ^ οιμ ^ there are three photographs: one showing both outbreaks, and one separately for each outbreak.

Based on the interview with the patient, the itching is assessed using the following scale:

= none = minimal: accidental scratching = moderate: frequent scratching = severe: frequent scratching, may disturb sleep.

At the end of the study, a cumulative assessment of response to treatment is made based on the following criteria:

- no trace = 100% reduction

- significant improvement = varied LPSI format by at least 6656 and / or varied focus area by at least 66% cr

- moderate improvement = decrease in LPSI r 65-3356 and / or decrease in the area of the focus r 65-33%

- bad: loss of LPSI r less than 32% and / or less than 32% of the focus area

- worse: exacerbation of the disease

Therapeutic success occurs with CEaE !, when there is no trace of any of the outbreaks, or when there is significant improvement.

155 712

The doctor and patient "will also delete which side, left or right, has been better treated according to the following criteria:

- the left side is definitely better

- left side slightly better

- no detectable difference

- the right side is slightly better

- the right side is much better

V To test post-treatment efficacy, LPSI scores and individual focal areas are compared using the WIŁCOXON-Sign Rank test for paired treatment between cyclosporin-treated and placebo-treated sites with respect to zMan in the assessment between baseline and baseline size. after 4 weeks of treatment.

After all, further comparisons / concerning other moments of time and other parameters of effectiveness / along with related values are taken into account in the description of the progress by the relationship.

In the above-mentioned α trials of subjects, there is a significant improvement in the condition of the sclerosis site, i.e., LPSI size for erythema, desquamation and skin thickness, on the sites where the test agent containing cyclosporin was applied, compared to the sites treated with placebo. An improvement in the subject score was also observed along with an improvement in the itch scores recorded on the atron treated with oyclosporin compared to the side treated with placebo.

The results were also obtained in trials conducted according to the parallel formula on individuals with contact dermatitis, based on three or more of the following criteria:

- itching

- types of morphology and distribution of: swelling or linearity in adults, attacks of the face and extension muscle in infants and children

- chronic or chronically relapsing dermatitis

- a personal or family history of atopy / asthma, α-lung rhinitis, atopic dermatitis / and three or more ceoh αbotznyoh, which include: conjunctival dryness, ichthyosis / palmate keratosis / keratosis, immediate skin test / type 1 /, elevated serum IgE levels, early age of disease attack / less than 5 years /, susceptibility to skin infections / especially itaph. aureus and Herpes simplex / / decreased cell-dependent immunity, tendency to non-specific dermatitis on the hands or feet, eczema on the nipples, cheilitis, recurrent conjunctivitis, under the orbital Dennie-liorgan fold, conical cornea, anterior subcapsular cataract, darkening of the orbit, facial pale / facial erythema, white dandruff, frontal cervical fold, itching after eating sweets, intolerance to wool and lipad solvents, peri-bellows contrast, food low-oxygenation, environmental and emotional factors influencing the course of the disease, as well as skin whitening and delayed fading. The test is carried out for 3 weeks, assessing the following parameters on a scale from 0 to 3: itching, erythema, exudate, unpleasant sensations and lichen.

Topical compositions containing 1-1055 cyclosporins, e.g., a 5% agent, have been found to be effective in the treatment of allergic contact dermatitis as well as in the treatment of alopecia.

The method of administration of cyclosporin or to carry out treatment with cyclosporine subject passing such a treatment or wyiamasi-limiting of such treatment is ηβ that is administered to said subject an effective amount of cyclosporin in the orthorhombic crystal form, in particular in non-solvated orthorhombic crystalline form as CM / JOIII, for example. Zde ^of in ^it ÓΓym ^and olaie ^k with points AA ?.

155 712

Specifically, the method of administering immunosuppressive or anti-inflammatory therapy in a subject in need of such treatment comprises administering to the subject an effective amount of cyclosporin in an orthorhombic crystalline form, in particular an unsolvated orthorhombic crystal form such as CY-I / X-III, e.g. as defined in any one of points / - /.

The scope of the above methods includes the treatment of any of the diseases and conditions described above in relation to the use of cyclosporine, e.g., prevention of transplant rejection, treatment of jutrimental or rheumatic disease, treatment of any other disease or condition in which there is an undesirably strong immune system response. treatment of parasite infections, treatment of resistance to chemotherapeutic agents, e.g. reducing tumor resistance to anti-cancer chemotherapy, and treatment of alopecia. The foregoing pups are particularly useful in the treatment of skin or eye diseases or conditions, e.g., psoriasis, atopic or contact dermatitis, in corneal transplantation, in the treatment of keratitis by intrauterine administration, and in the treatment of rheumatoid arthritis or other diseases or disease states of the joints by intra-articular administration.

The cyclosporin of the invention in an unsolvated orthorhombic crystalline form such as CY-I / Σ-ΙΙΙ, e.g. as defined in any of items / - / above, is used as a pharmaceutical, e.g. for diseases or conditions, e.g. for use as an immuno-oppressive or anti-inflammatory agent.

Cycle in orthorhombic crystalline ptgti such as CY-A / Y-III, e.g. as defined in any of items (-) above, is also used in the preparation of pharmaceutical agents for the purposes of the above-mentioned term.

/ to facilitate the identification of cyclosporine in the orthorhombic crystal form, in particular Cy-Α / Χ-ΙΙΙ, the following tables I-III provide X-ray powder diffraction data for Cy-Α / Σ-ΙΙΙ, as well as for two other forms, Cy- / / XI and Cy-A / X-II. Figure 4 shows the X-ray powder diffraction programs from which the data shown in Tables 1-3 were obtained. X-ray images were obtained with the quinier-DeWolff II apparatus, with w / y ^ kO ^ y ^ EJtanl ^ ^ m CuK ° Co aC = 0.1542 nm radiation.

Table 1 Cy-Λ / - I Dphiractogram Line no d / nm / Rat gosh 1 1.30 VS 2 1.14 VS 3 1.03 VS 4 0.97 VS 5 0.94 VS 6 0.825 vs 7 0.71 s 8 0.61 M. 9 0.585 s 10 0.56 s 11 0.525 vs 12 0.481 s 13 0.458 s 14 0.425 M. 15 0.40 AT 16 0.367 at 17 0.345 M.

155 712

Line number 1 2

9

Table 2 Cy-A / X-II X-ray radiograph

d / nm / Intensity 1.79 S. 1.14 VS 1.10 S. 1.06 AT 1.02 s 0.89 M. 0.87 0, S1 N 0.72 VS 0.63 AT 0.595 H. 0.57 H. 0.535 AT 0.51 s 0.485 s 0.48 at 0.455 at 0.44 at

Table 3 Cy-A / X-III diffractogram

.nia no d / nm / Intensity 1 1.20 AT 2 1.04 vs 3 0.96 s 4 0.87 s 5 0.79 at 6 0.77 s 7 0.67 at 8 0.60 at 9 0.583 s 10 0.53 M. 11 0.52 M. 12 0.492 s 13 0.488 s 14 0.458 at 15 0.448 at 16 0.40 at 17 0.359 at 18 0.338 at

VS = very strong S = strong M = medium

155 712

Claims (1)

Patent claims 1. Method for the production of cyclosporin in a new nlesolvated orthorhombic crystal form, lattice t a = 1.27 nm, b = 1.57 nm, c = 3.63 nm and a triclinic unit volume of 180.4 nm and having the following X-ray diffraction characteristics: The μ line d / nm / No, no 1 1.20 medium 2 1.04 very strong 3 0.96 strong 4 0.87 strong 5 0.79 medium 6 0.77 strong 7 0.67 medium 8 0.60 medium 9 0.583 strong 10 0.53 medium 11 0.52 medium 12 0.492 strong 13 0.488 strong 14 0.458 medium 15 0.448 medium 16 0.40 medium 17 0.359 medium 18 0.338 medium with the fact that cyclosporin dissolves in a solvent medium polymer with a molecular weight of co at least 200 containing non-volatolates rhombic crystal of cyclosporine, suitably in a polymeric ether of partial weight at least 200 at temperature best of all 40 ° C, then the environment is cooled and from gaining crystalline cyclosporine. 2. The method according to p. 1, with n a m and e However, a solvent medium containing polyethylene glycol or polyptopylene glycol is used. FiG. 1 0.1% oral solution - ° ^- cyclosporin - <- Placebo Time after 2nd DNFB Challenge Prebie (hrs) 155 712 FiG. 2 FiG. 3 0.1% cyclo-spore ointment as in Example 4 5 ~ o ~ Placebo Time after 2nd Challenge DNFB (hrs) —MeBmt-q (Abu-Sar-MeLeu-VaI-MeLeu-Ala- (D) Ala-MeLeu-MeLeu-MeVal - 1 2345 678 9 10 11 PATTERN 155 712 Fig. IV Cy-A / X-I Cy-A / X-II Cy-A / X-III Department of Publishing of the UP RP. Circulation of 90 copies