LU87613A1 - NOVEL CYCLOSPORIN-BASED OPTHALMIC COMPOSITIONS - Google Patents

Description

The present invention relates to new ophthalmic compositions, that is to say compositions intended for application to the eye, which comprise a cyclosporine as active substance and which are suitable for the treatment of diseases and conditions of the eye. eye and surrounding areas.

Cyclosporins represent an extended class of peptides with valuable pharmacological properties, for example immunosuppressive, anti-inflammatory, and / or anti-parasitic activity and / or activity intended to suppress the resistance of tumors to treatments with antineoplastic or cytostatic agents . Cyclosporins include, for example, fungal metabolites of natural origin, such as cyclosporins A, B, C, D and G, as well as a wide variety of synthetic or semi-synthetic cyclosporins, for example dihydro- and iso-cyclosporins (see, for example, U.S. patents 4,108,985, 4,210,581 and 4,220,641), [(D) -Ser] 8-Ciclosporin (see U.S. patent 4,384,996), [0-acetyl- ( D) Ser] 8-Ciclosporin (see US Patent 4,764,503), [ß-fluoro- (D) Ala] 8-Ciclosporin (see British Patent Application 2,206,119A), [Val] 2 - [(D) methyl-thio-Sar] 3- and [Dihydro-MeBmt] 1- [Val] 2 - [(D) methylthio-Sar] 3-Ciclosporin [see US Patent 4,703,033] and many still others.

Among the cyclosporines, the one which has so far been subjected to a greater number of researches is cyclosporin A, also known as Ciclosporin and available on the market under the brand SANDIMMUN or SANDIMMUNE. Ciclosporin has been shown to selectively suppress a variety of T cell functions including prevention of maturation and expression of sensitized T cells in cell mediated immune responses, and Ciclosporin is widely used today to suppress rejection in organ transplants. Ciclosporin has also been used systematically in the treatment of autoimmune or inflammatory intraocular diseases such as uveitis. However, due to the side effects associated with systemic treatment, Ciclosporin has had limited use in the treatment of such eye conditions.

Effective administration of Ciclosporin to the eye by the topical route would make it possible to greatly reduce or eliminate systemic side effects by limiting activity to the area to be treated and proposals to this effect have been made (see for example the patent American 4,649,047). However, the usefulness and effectiveness of Ciclosporin in the treatment of diseases and conditions of the eye have been neglected by the absence of an appropriate composition which is acceptable or effective for use topically. There is therefore a need for a composition which does not cause discomfort for the patient and which allows practical administration, that is to say which does not require too frequent administration while providing an appropriate supply of the drug to external regions and, in particular, to the internal regions of the eye.

Similar considerations apply to other cyclosporins known to those skilled in the art, for example those proposed for use as immunosuppressive or anti-inflammatory agents, for example cyclosporin G, also known and hereinafter designated [Nva] 2-Ciclosporin.

The present invention relates to new ophthalmic compositions comprising a cyclosporine as active substance, which overcome the difficulties encountered in the state of the art, for example of the type indicated above. The compositions of the invention are intended for topical application, that is to say application to or in the eye, for example on the cornea or on the corneal epithelium, or in the surrounding areas. immediately the eye, for example on the internal surfaces of the upper or lower eyelid.

Applied topically as indicated above, the compositions of the invention are useful for the treatment of diseases or conditions of the eye or of the organs or tissues which surround or are associated with it, for example the glands and channels lacrimaux, especially diseases and immunological or inflammatory conditions. The compositions of the invention are more particularly useful for the treatment of diseases or conditions involving an excessively high immune response or inflammatory reaction or process as a component or part of their etiology, in particular autoimmune diseases of the eye . The diseases and conditions which can be treated by the compositions of the invention include, for example, uveitis (anterior and posterior), chronic keratitis, conjunctivitis (including in particular spring conjunctivitis), spring keratoconjunctivitis, dry keratoconjunctivitis and keratoplasty (that is to say in the case of a corneal transplant) as well as inflammatory or immunological conditions caused by eye surgery in general. The compositions of the invention can also be used for the induction or the maintenance treatment of tears, for example when the lacrimal function is reduced, for example following any disease or condition indicated above.

It has surprisingly been found that the ophthalmic compositions of the invention cause little or no irritation or discomfort in the patient and have a therapeutic effect at appropriate application rates in the corneal areas and in the internal areas of the eye. the eye, including the anterior chamber, the posterior chamber, the vitreous body, the aqueous humor, the vitreous humor, the cornea, the iris / ciliary body assembly, the lens, the choroid / retina assembly, the sclera or the organs or tissues surrounding the eye, for example the lacrimal duct or the lacrimal gland.

According to a first aspect, the invention relates more particularly to: A) an ophthalmic composition comprising a cyclosporine as active substance and "as a vehicle, (1) a vegetable oil acceptable from the ophthalmic point of view and (2) an acceptable petrolatum of the ophthalmic point of view.

The compositions of the invention are intended for topical ophthalmic application, that is to say an application to the surface of the eye, for example on the cornea or the epithelium of the cornea, or on the areas surrounding the eye.

The term "ophthalmically acceptable" means that the substance is suitable for or allows application to the eye, for example that it is safe for ophthalmic application topically at the doses to be administered.

The preferred cyclosporins for use in the compositions of the invention are cyclosporins having immunosuppressive or anti-inflammatory activity, for example as described above, in particular Ciclosporin. Another preferred cyclosporin for use in the compositions of the invention is [Nva] 2-Ciclosporin. The compositions of the invention are suitable in the form of an ointment comprising the components (1) and (2) defined above, as basic components of the ointment.

Cyclosporine, for example Ciclospori-ne, is advantageously present in the compositions of the invention in an amount of between approximately 0.01 and approximately 10% (unless otherwise indicated, all the percentages indicated in the description and in claims s '' mean by weight relative to the total weight of the composition). Cyclosporine is preferably present in an amount between about 0.05 and about 10%, especially between about 0.05 and about 5%, preferably between about 0.1 and 2.5%. Cyclosporine is more preferably present in an amount between about 0.1, 0.5, 1.0 or 2.0%.

Component (1) (vegetable oil] is advantageously present in the compositions of the invention in an amount of at least 25%. Component (1) is suitably present in an amount between about 25 and about 65%, preferably between about 25 and about 45%, more preferably between about 35 and about 45% and especially between about 40 to about 45%.

Component (2) [petroleum jelly] is advantageously present in the compositions of the invention in an amount of at least 25%, preferably at least 50%. Component (2) is suitably present in an amount between about 25 and about 65%, preferably between about 50 and about 65%, more preferably between about 50 and about 60%, and especially between about 50 and about 55 %.

Components (1) and (2) are suitably present in the compositions of the invention in a weight ratio of about 1: 2 to 2: 1, for example about 1: 1.

Component (1) can include any suitable vegetable oil, for example olive oil, peanut oil, corn oil, castor oil or sesame oil, or mixtures thereof. However, component (1) preferably comprises corn oil, for example as described in the Handbook of Pharmaceutical Excipients ("ΗΡΕ"), a joint publication of the American Pharmaceutical Association and the Pharmaceutical Society of Great Britain, on page 91.

Component (2) can comprise any suitable petrolatum, for example those available under the names of white petrolatum (white petrolatum, USP; USP = * American pharmacopoeia), white soft paraffin (white soft paraffin, BP / EP; BP / EP " British and European pharmacopoeias), white petrolatum gel (white petroleum jelly) or petrolatum gel (petroleum jelly) or under the names of yellow petrolatum (petrolatum yellow, USP), yellow soft paraffin (yellow soft paraffin, BP / EP), yellow petroleum jelly or yellow petroleum jelly - for additional information see HPE, pages 194-195. As component (2), white petrolatum (white petrolatum) is especially preferred, for example of one color (maximum, Lovibond Color device - 2 "cell, ie 5.08 cm) = approximately 0.5 to 18Y (yellow intensity) , 0.5R (red intensity).

Advantageously, the compositions according to the invention also comprise (3) an emulsifier. As suitable component (3), mention may be made of any emulsifier acceptable from the ophthalmic point of view, for example any emulsifier known to specialists. The preferred emulsifiers for use in the compositions of the invention are nonionic emulsifiers, in particular nonionic lanolin derivatives or extracts, especially lanolin alcohols (see HPE, page 164) or in general the 7 emulsifiers with sterol base. As examples of emulsifiers suitable for use in the compositions of the invention, mention may be made of the products known and marketed under the brand Amerchol, in particular Amerchol 400, C, CAB, H9, L-99, 4-500 and RC and especially the Amerchol CAB. [For a more complete definition of the above products, see Fiedler, "Lexikon der Hilfstoffe, 3rd edition (1989), pages 138-139].

When the components (3) are used in the compositions of the invention, they are advantageously present in an amount of between approximately 0.5 and approximately 10%, preferably between approximately 0.5 and approximately 5%, more preferably between approximately 1 and 2.5%, for example around 2%.

The compositions of the invention may also include any other component suitable for use in ophthalmic preparations, for example preservatives or anti-microbial agents. In particular, they include (4) a preservative, chloro-butanol (for example anhydrous chlorobutanol) - see HPE, pages 72-73 - being particularly suitable. Such additional components, for example components (4), when used, are advantageously present in an amount of up to about 5.0%, more preferably up to about 2.0%, e.g. from about 0.01 to about 2.0% or up to about 5.0%, preferably from about 0.1 to about 1.0%, for example from about 0.5%. The compositions of the invention can therefore advantageously comprise, for example, the following ingredients in the relative amounts indicated: CYCLOSPORINE [for example CICLOSPORINE OR [Nva] 2-CICLOSPORINE] 0.01 to 10% (1) 25 to 65% ( 2) 25 to 65%, preferably 50 to 65% (3) 0.5 to 10% (4) 0.1 to 5% plus any other component to reach 100%, components (1) and (2) preferably being present in a weight ratio of about 1: 2 to 2: 1, and the preferred amounts of the individual components can be independently selected from those indicated above. The following example illustrates the invention without in any way limiting its scope.

EXAMPLE

COMPONENTS

CYCLOSPORINE (e.g. the

Ciclosporin) 2.0% (1) Corn oil 41.65% (2) White petrolatum 53.9% (3) Non-ionic lanolin derivatives (e.g. AMERCHOL CAB) 1.96% (4) CHLOROBUTANOL ( anhydrous) 0.49% TOTAL 100.00%

Equivalent compositions comprising 0.1, 0.5 and 1% cyclosporine (e.g. cyclosporine) can be prepared by increasing each of components (1) to (4) by the proportional amount required.

The composition, which is in the form of an ointment, can be prepared according to the usual techniques, for example by adding (1) to (2) and heating to about 35 to 60 ° C to form a clear liquid, and then dissolution of (3) and (4) with stirring in the clear liquid to form the vehicle intended for the ointment. Ciclosporin is then dissolved with stirring in the liquefied vehicle, the resulting scrub being ready for final packaging. It is also possible to dissolve Ciclosporin in component (1) or in a portion of component (1) and then add the solution obtained to the other ingredients before packaging. For production in large quantities, Ciclosporin is advantageously dissolved in the mixture of components (1) - (4) at a high temperature using a mixer with high shear forces. Under sterile conditions, the solution obtained is then filtered and poured into containers, for example tubes for ointment, advantageously containing up to approximately 5 g, for example 3.5 g of ointment per container.

During their preparation, the compositions of the invention are preferably filtered, for example under sterile conditions, at the end of the dissolution of the cyclosporine and before proceeding to packaging.

For use in therapy, the compositions of the invention are preferably poured into suitable containers, for example intended for direct ophthalmic application by topical content, for example ointment tubes as indicated above having a closure system of a relatively small diameter and allowing easy application of the content to the surface of the eye, for example in an amount between approximately 0.05 and approximately 0.2 ml, for example between approximately 0.05 and approximately 0, 1 ml.

The activity of the compositions of the invention can be demonstrated by tests on animals, for example as described below, or by clinical trials.

TEST METHOD

The objective of the test is to determine the penetration into the rabbit eye and the radioactivity reservoirs of the various tissues of the rabbit eye as well as the systemic concentrations following a single topical application of the compositions comprising Tritiated ciclosporin ("Ci-3H"), looking for the distribution of Ci-3H in the tissues of the eye and the levels of Ci-3H obtained in the blood (systemic concentrations).

According to the usual techniques, Ci-3H is prepared by introducing tritium into the amino acid residue 1 (-MeBmt-) of Ciclosporin, for example as described by Voges et al., In "Synthesis and Applications of Isotopically Labeled Compounds ", Ed. RR Muccino, Elsevier Press Amsterdam, 371-376 (1986), in the article" Tritiated Compounds for In-Vivo Investigation ". The purity and the identity of the products are determined by NMR spectra, thin layer chromatography / thin layer radio chromatography and reverse phase HPLC. For the test, CI-3H and unlabeled cyclosporine are used in a dilution ratio of 1:10.

For the test, female white rabbits from New Zealand (Lab Rab Co.) weighing 4 to 6 kg are used. The animals are placed in cages where they receive standard food (Purina Chow) and unlimited water. For approximately 1 hour after application of the composition to be tested, the rabbits are left in a normal vertical position in an apparatus which prevents them from moving. The compositions to be tested are administered at doses of 20 μΐ or 19.9 mg using a graduated pipette.

The pipettes are filled with care in order to avoid air bubbles and the composition to be tested is carefully expelled into the lower part of the conjunctival cul-de-sac in both eyes of the rabbits. The eyelids are held together for 1 second and released. Eyes are visually checked for any signs of irritation / tears caused by irritation or cloudiness according to the Draize test modified by Calgon using a dissection lamp instead of a slit lamp (Draize, jH, Appraisal for the Safety of Chemicals in Foods, Drugs and Cosmetics, association of Food and Drug Officials of the United States, Texas State Department of Health, Austin, Texas, 1959, method modified by Calgon Consumer Products Research Labs, Toxicology, 1973).

After application of the dose, the rabbits are sacrificed in groups of 3, after 0.3, 1, 2, 4, 6, 12, 24, 48 and 96 hours. Just before the animals are sacrificed, blood samples are taken from the marginal ear vein. The animals are then sacrificed by injection of a T-61 euthanasia solution (Hoechst) into the marginal vein of the ear. Both eyes are rinsed with normal physiological solution (0.9% NaCl) and enucleated without rupturing the surrounding blood vessels. The ocular surface is carefully rinsed and dried with filter paper to remove any remaining medicine from the tear fluid. The aqueous humor is removed using a 3/8 inch (9.52 mm) 26 gauge needle attached to a 0.2 ml pipette inserted into the corneal limbus. The cornea is incised at the limbus and the iris and ciliary body are dissected as a single sample. The lens is removed and an aliquot of the vitreous humor is collected. The excess vitreous humor is removed from the remaining tissues using filter paper. The choroid / retine assembly is removed by scraping from the sclera and a sclera sample is taken from the area closest to the optic nerve. To avoid contamination, clean instruments are used to dissect individual tissues. Each sample is rinsed, dried on filter paper, placed in a combustion dish and weighed. Blood samples (* s 200 μΐ) are also placed in combustion cups and weighed. The tissue and blood samples are air dried before combustion in a Packard Tri-Carb R Sample Oxidizer, model 306 using a Monophase E -40 (Packard Instrument Co.). After combustion, the radioactivity of the samples is determined using a Packard Tri-Carb H, model 460 liquid scintillation spectrometer.

To determine the specific activity, an aliquot of 2.0 μΐ of the composition to be tested is dissolved in 100 ml of tert.-butyl methyl ether and 0.1 ml of the solution is analyzed according to the combustion method. ethereal. Using the specific activity, starting from the disintegrations per minute per gram (dpm / g), the concentration of radioactivity in the tissues is converted into equivalent nanograms of Ciclosporin / gram of tissue or blood.

The radioactivity of all the samples is determined within the limit of the statistical error of 7% with a confidence limit of 95%. This implies that any count per minute (cpm) less than 3 is not significantly different from zero and corresponds to a detection limit of 34 picograms per gram (pg / g) ·

During a series of tests carried out according to the above method, the following compositions were compared: TEST COMPOSITION A:

Composition of Example 1 according to the present invention.

TEST COMPOSITION B:

Comparative composition comprising Ciclosporin and components (2) to (4) in amounts identical to those of Example 1 and using as component (1) 41.65% mineral oil instead of l corn oil.

TEST COMPOSITION Ci

Comparison composition, comprising 2% Ciclosporin and 98% castor oil.

(to pour composition B into pipettes, the composition is first heated to 30 ° C to facilitate the operation and then allowed to cool to room temperature before application). RESULTS

Figures 1 to 7 attached relate to the graphic representations of the variations, as a function of time, of the concentration of radioactivity in each tissue of the eye analyzed. On each graph, the time in hours is plotted on the abscissa up to 96 hours; the concentration in mg Eq of Ciclosporin / gram of tissue is plotted on the ordinate, all the values being normalized to a dose of 0.335 mg of Ciclospori-ne / eye.

The results for composition A are represented by black circles.

The results for composition B are represented by black squares.

The results for composition C are represented by the inverted triangles.

Fig. 1 gives the results for the aqueous humor.

Fig. 2 gives the results for the cornea.

Fig. 3 gives the results for the whole iris / ciliary body.

Fig. 4 gives the results for the lens.

Fig. 5 gives the results for the vitreous humor.

Fig. 6 gives the results for the choroid / retina assembly.

Fig. 7 gives the results for the sclera.

Figures 1 to 7 indicate a relatively rapid initial absorption of Ciclosporin in the anterior and posterior tissues of the eye with the three formulations. However, the maximum concentrations in the cornea (fig. 2), which seems to serve as a reservoir for the other intraocular tissues, were obtained approximately 3 times faster with composition A (approximately 2 hours) than with composition B (about 6 hours). In addition, the concentrations obtained in the cornea with composition A are much higher than those obtained with composition B. Composition C requires approximately 12 hours to reach a maximum concentration in the cornea.

The resultant and surprising increase in the intake of substance in the deepest tissues of the eye is confirmed by the results on the aqueous humor (fig. 1) where the area under the curve (AUC) obtained with the composition A for 96 hours is approximately 2 times greater than that obtained with composition B and 3 times greater than that obtained with composition C. [the values of the area under the curve (96 hours) (mg Eq / g) normalized at 0.335 mg / eye are the following: 636 for composition A; 334 for composition B; 188 for composition C]. This advantage of composition A is confirmed for the whole iris / ciliary body (fig. 3) and the lens (fig. 4).

It is therefore found that composition A is clearly superior to compositions B and C as regards the supply of substance in the anterior zones of the eye, in particular the cornea and the aqueous humor / iris / ciliary body unit. The compositions of the invention are therefore particularly advantageous for the treatment of diseases or conditions of the eye affecting these areas, for example for corneal transplants, and the treatment of uveitis and dry keratoconjunctivitis.

With regard to the posterior segment of the eye, while the superiority of composition A is less marked for the vitreous humor and the sclera (figs. 5 and 7), it is notably superior for the choroid / retina group. (Fig. 7), these areas of the eye being more involved in autoimmune diseases affecting the posterior segment of the eye, alone or with other areas of the eye. Thus, composition A provides a significantly higher immediate supply as well as a higher and improved total total supply to the choroid / retina assembly. Thus, the value of the area under the curve (96 hours) for the choroid / retina unit (mg Eq / g normalized to 0.335 mg / eye) is 527 for composition A and only 296 for composition composition B and 184 for composition C.

Compared with compositions B and C, composition A is therefore surprisingly clearly superior in terms of supply of substance to the posterior segment of the eye, comprising the choroid / retina assembly, and therefore has a particular advantage for the treatment of eye diseases and conditions affecting these areas, alone or with other areas, for example for the treatment of posterior uveitis and other retinopathies of immunological or inflammatory origin.

As is apparent from FIGS. 1 to 7, composition C exerts in all cases an activity clearly lower than that of compositions A and B.

The following table shows the systemic concentrations of Ciclosporin in the blood (ng Eq / g normalized to 0.335 mg / eye).

As can be seen from the results obtained, the systemic levels measured are relatively low for the three compositions tested, but surprisingly more particularly for composition A compared to the other two. Composition A thus appears clearly superior in terms of contribution of active substance to the eye and its tissues as well as systemic non-involvement.

A visual observation of the eye treated with composition A according to the modified Draize test, did not make it possible to note an irritation linked to the composition.

The advantageous therapeutic properties of the compositions of the invention can also be demonstrated by clinical trials, for example by administration to patients suffering from the diseases or conditions of the eye indicated above.

For such tests and for practical therapeutic use, for example for the treatment of uveitis (anterior or posterior), conjunct spring tivitis, kerato-conjunctivitis spring or kerato-conjunctivitis sicca, the compositions of the invention, for example the composition of example 1, are advantageously administered on or in the eye in individual amounts for example of approximately 0.1 to 0.2 ml 1 to 4 times per day, for example depending on the particular disease or condition to be treated, the clinical condition and the desired effect. By continuing the treatment, there is a marked improvement, for example compared to the untreated controls, for example over a period of 1 to 2 weeks or more. The compositions of the invention are well tolerated by patients undergoing treatment and do not cause significant or unpleasant irritation.

In accordance with the above, the invention also relates to: B) A composition as defined under A) for use in the treatment of diseases or conditions of the eye or of the organs or tissues surrounding the eye or which therein are associated, in particular for the treatment of diseases or immunological or inflammatory conditions of the eye or of the organs or tissues surrounding the eye or which are associated therewith, the zones, the segments or the particular tissues of the eye being such as defined above and the diseases or conditions of the eye being as defined above.

Claims (12)

1. An ophthalmic composition, characterized in that it comprises a cyclosporine as active substance and, as a vehicle, (1) a vegetable oil acceptable from the ophthalmic point of view and (2) a petrolatum acceptable from the ophthalmic point of view. 2. A composition according to claim 1, characterized in that the cyclosporine is Ciclosporin or [Nva] 2-Ciclosporin. 3. A composition according to claim 1 or 2, characterized in that the cyclosporine is present in an amount between about 0.01 and about 10%. 4. A composition according to claim 3, characterized in that the cyclosporine is present in an amount between about 0.1 and about 2.5%. 5. A composition according to any one of claims 1 to 4, characterized in that the component (1) is present in an amount between about 25 and about 65%. 6. A composition according to any one of claims 1 to 5, characterized in that the component (2) is present in an amount between about 25 and about 65%. 7. A composition according to claim 6, characterized in that the component (2) is present in an amount between about 50 and about 65%. 8. A composition according to any one of claims 1 to 7, characterized in that the weight ratio of the component (1) to the component (2) is between approximately 1: 2 and 2: 1. 9. a composition according to any one of claims 1 to 8, characterized in that the component (1) comprises olive oil, peanut oil, castor oil, l sesame oil or corn oil. 10. A composition according to any one of claims 1 to 9, characterized in that it also comprises (3) an emulsifier acceptable from the ophthalmic point of view and (4) a preservative. 11. A composition according to claim 10, characterized in that the component (3) is present in an amount between approximately 0.5 and approximately 10% and the component (4) is present in an amount which can range up to approximately 5.0%. 12. A composition as specified in any one of claims 1 to 11, for use in the treatment of diseases or immunological or inflammatory conditions of the eye or of organs or tissues surrounding the eye or which are therein. associated.