Ocular compositions

Abstract

The invention provides pharmaceutical composition comprising compound of formula (1) and L-Histidine, and use thereof.

Description

OCULAR COMPOSITIONSBACKGROUND OF THE INVENTION

[0001] L-tryptophan is an essential heterocyclic amino acid which is part of various naturally occurring peptides. Tryptophan forms a large variety of small peptides. Many of these peptides have physiological activities. Some small peptides (such as Trp-Pro) are under development for the treatment of diseases, e. g. for amyloid-associated diseases, see US 2006 / 0234947. However, pharmaceutical compositions containing amino acid derivatives often are not stable during storage, in particular when formulated as aqueous solutions. For many pharmaceutical applications, the amount of amino acid derivative needed at the specific site of action is higher than conventional pharmaceutical formulations can provide with.

[0002] US 15 / 848,044 discloses tryptophan derivatives, such as compound of formula (1) and compositions comprising them.

[0003] However, these compounds have shown very low solubility in water and organic solvents and have a limited stability as these compounds can cyclize to a diketopiperazine. In addition, most salts of compound of formula (1) are amorphous or glassy (sulfate, hydrogensulfate, phosphate, citrate, mesylate, etc.). The physical state presents difficulties in adjusting for pharmaceutical applications.

[0004] The compound of Formula (1) is limitedly soluble at pH 6-8, which is applicable for ocular administration, and presents sufficient solution stability only in higher pH, which cannot be administered into the eye.

[0005] W02012010415 describes formulations which contain solubility enhancers like cyclodextrines to improve the stability of such compounds. However, solubiliy enhancers are sometimes irritation and not optimal for chronic use.

[0006] Thus, there is a need to find a solution for improving the solubility at physiological pH, and stability of compound of formula (1) in order to allow the pharmaceutical composition to be administered safely, well-tolerated, and efficiently in a proper dose and administration mode to a patient in need thereof.SUMMARY OF THE INVENTION

[0007] In some aspects, disclosed herein is a pharmaceutical composition comprising compound of formula (1)and L-histidine.

[0008] In some related aspects, the pharmaceutical composition comprises a salt of compound of formula (1).

[0009] In some related aspects, the pharmaceutical composition comprises a solvate salt of compound of formula (1), wherein said solvate is C3 or C4 alcohol thereof.

[0010] In some further related aspects, the solvate salt of compound of formula (1) is a crystalline solvate salt.

[0011] In some related aspects, the pharmaceutical composition further comprises citric acid, citrate or combinations thereof.

[0012] In some further related aspects, the pharmaceutical composition has pH in the range of 5-7. In some aspects, the pharmaceutical composition has pH in the range of 6.3-6.7.

[0013] In some further related aspects, the molar ratio of compound of formula (1) and L-histidine is in the range 1.4-2.0. In some aspects, the molar ratio of compound of formula (1) and L-histidine is in the range 1.70-1.73.

[0014] In some further related aspects, the composition is an aqueous composition, wherein compound of formula (1) is in a concentration in the range of 1 mg / ml to 50 mg / ml. In some aspects, compound of formula (1) is in a concentration of 20 mg / ml.

[0015] In some further related aspects, the pharmaceutical composition is suitable for ocular administration. In some aspects, the ocular administration is by eye drops, eye creams, eye sprays, intraocular depot formulations, by injection including intraocular injection, intra- or periocular injections, comprising intraocular fluids, intravitreal injections, injections into the supra-arachnoidal or the subretinal space, depot formulations, solid and semisolid intraocular carriers and matrices, or ocular devices.

[0016] In some aspects, disclosed herein is a pharmaceutical composition as described herein, for the treatment of an ophthalmic disorder or disease. In some related aspects, the ophthalmic disease or disorder comprises amyloid-associated retinal disease.

[0017] In some aspects, disclosed herein is a process for preparation of a pharmaceutical composition, comprising compound of formula (1)and L-histidine, comprising the steps of dissolving, compounding, filtering and filling the aqueous solutions into primary packaging materials.BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The subject matter regarded as the invention is particularly pointed out and distinctly claimed in the concluding portion of the specification. The invention, however, both as to organization and method of operation, together with objects, features, and advantages thereof, may best be understood by reference to the following detailed description when read with the accompanying drawings in which:

[0019] Fig. 1 is a graphical illustration of six formulations that were compounded by addition of 5 mg / ml glycerol and 5 mg / ml HPMC in water, adding 50 mg / ml of the solubilizer and excess of GAL-101 salt and adjusting the pH with citric acid solution to a pH of 6.0. The suspension was equilibrated for 24 hours at room temperature and then filtered through a 0.45 pm filter. Afterwards the content of GAL-101 was determined by HPLC and solubility calculated;

[0020] Fig. 2 is a graphical illustration of four formulations that were compounded by addition of 5 mg / ml glycerol and 5 mg / ml HPMC in water, adding different buffer types each in a concentration of 100 rnM to the solution and then an excess of GAL-101 salt and adjusting the pH with hydrochloric acid solution to a pH of 6.0. The suspension was equilibrated for 24 hours at room temperature and then filtered through a 0.45 pm filter. Afterwards the content of GAL-101 was determined by HPLC and solubility calculated;

[0021] Fig. 3 is a XRDP presentation of Form A;

[0022] Fig. 4 is a XRDP presentation of Form B;

[0023] Fig. 5 is a XRDP presentation of Form C;

[0024] Fig. 6 is a XRDP presentation of Form D;

[0025] Fig. 7 is a XRDP presentation of Form E.

[0026] It will be appreciated that for simplicity and clarity of illustration, elements shown in the figures have not necessarily been drawn to scale. For example, the dimensions of some of the elements may be exaggerated relative to other elements for clarity. Further, where considered appropriate, reference numerals may be repeated among the figures to indicate corresponding or analogous elements.DETAILED DESCRIPTION OF THE PRESENT INVENTION

[0027] In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the invention. However, it will be understood by those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the present invention.Composition

[0028] In some embodiments, disclosed herein is a pharmaceutical composition comprising compound of formula (1)and L-histidine.

[0029] In some embodiments, the pharmaceutical composition comprises a salt of compound of formula (1). In some embodiments, the salt is selected from Hydrochloride salt, acetate salt, succinate salt, potassium salt and sodium. In one embodiment, the salt is a Hydrochloride salt. In another embodiment, the salt is acetate salt. In another embodiment,the salt is a succinate salt. In another embodiment, the salt is a potassium salt. In another embodiment, the salt is sodium salt.

[0030] In some embodiments, the pharmaceutical composition comprises a solvate salt of compound of formula (1), wherein said solvate is C3 or C4 alcohol thereof.

[0031] In some embodiments, the pharmaceutical composition comprises a crystalline form of a solvate salt of compound of formula (1), wherein said solvate is C3 or C4 alcohol thereof.

[0032] In some embodiments, the salt is an alkali salt.

[0033] In some embodiments, the alkali salt is selected from sodium and potassium. In one embodiment, the alkali salt is sodium. In another embodiment, the alkali salt is potassium.

[0034] In some embodiments, the solvate is selected from 1-propanol, 2-propanol, 1- butanol, isobutanol and any combinations thereof. In one embodiment, the solvate is 1- propanol. In another embodiment, the solvate is 2-propanol. In another embodiment, the solvate is 1-butanol. In another embodiment, the solvate is isobutanol.

[0035] In some embodiments, the crystalline form of a solvate salt of compound of formula (1) are form A, form B, form C, formd D or form E, according to Table 1.

[0036] In one embodiment, the invention provides a sodium 2-propanol solvate of compound (1) having crystalline form A (the properties of are disclosed herein below and in Fig. 3).

[0037] In one embodiment, the invention provides a sodium 1-butanol solvate of compound (1) having crystalline form B (the properties of are disclosed herein below and in Fig. 4).

[0038] In one embodiment, the invention provides a sodium 1 -propanol solvate of compound (1) having crystalline form C (the properties of are disclosed herein below and in Fig. 5).

[0039] In one embodiment, the invention provides a sodium 2-propanol solvate of compound (1) having crystalline form D (the properties of are disclosed herein below and in Fig. 6).

[0040] In one embodiment, the invention provides a sodium 1 -butanol solvate of compound (1) having crystalline form E (the properties of are disclosed herein below and in Fig. 7).

[0041] Table 1 - solvated densities and of compound (1) for various solvate contents. Z is the number of formule units (molecules) per unit cell. V is the volume of the unit cell (A3). VO is the lattice volume for one formule unit (VO = V / Z, A3). Dx is the calculated density (from the volume and content of the unit cell). Dm is the measured density.

[0042] In some embodiments, the pharmaceutical composition further comprises citric acid, citrate or combination thereof. In one embodiment, the pharmaceutical composition further comprises citric acid. In another embodiment, the pharmaceutical composition further comprises citrate. In another embodiment, the pharmaceutical composition further comprises citric acid and citrate.

[0043] A skilled artisan would understand that citric acid is an organic compound with the chemical formula HOC(CO2H)(CH2CO2H)2. It is a colorless weak organic acid. It occurs naturally in citrus fruits. In biochemistry, it is an intermediate in the citric acid cycle, which occurs in the metabolism of all aerobic organisms. In the pharmaceutical industry, citric acid is used as an anti-chelation additive used in blood collection tubes and as a flavoring, pH chelator and preservative agent in various cosmetic and pharmaceutical products.

[0044] A skilled artisan would understand that citrate is the dissociated anion of citric acid. In the pharmaceutical industry, sodium citrate is used as an anticoagulant in blood transfusions and as a component of certain intravenous fluids. It is also used as a buffering agent to control the pH of medications and as a chelating agent to remove excess metals from the body.

[0045] In some embodiments, the pharmaceutical composition has a pH in the range of 5-8. In one embodiment, the pharmaceutical composition has a pH in the range of 5.5-6.9. In another embodiment, the pharmaceutical composition has a pH in the range of 5.8-6.8. In another embodiment, the pharmaceutical composition has a pH in the range of 6-6.7. In another embodiment, the pharmaceutical composition has a pH in the range of 6.1-6.7. In another embodiment, the pharmaceutical composition has a pH in the range of 6.2-6.7. In another embodiment, the pharmaceutical composition has a pH in the range of 6.3-6.7. In another embodiment, the pharmaceutical composition has a pH in the range of 6-7.

[0046] In some embodiments, the pharmaceutical composition has a pH of 5. In one embodiment, the pharmaceutical composition has a pH of 5.5. In another embodiment, the pharmaceutical composition has a pH of 6. In another embodiment, the pharmaceutical composition has a pH of 6.1. In another embodiment, the pharmaceutical composition has a pH of 6.2. In another embodiment, the pharmaceutical composition has a pH of 6.3. In another embodiment, the pharmaceutical composition has a pH of 6.4. In another embodiment, the pharmaceutical composition has a pH of 6.5. In another embodiment, the pharmaceutical composition has a pH of 6.6. In another embodiment, the pharmaceutical composition has a pH of 6.7.

[0047] In some embodiments, the molar ratio of compound of formula (1) and L- histidine is in the range 1.4-2.0. In one embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.5-2.0. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.6-2.0. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.7-2.0. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.8- 2.0. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.9-2.0. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.4-1.9. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.4-1.8. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.4-1.7. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.4-1.5. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is in the range 1.4-1.5.

[0048] In some embodiments, the molar ratio of compound of formula (1) and L- histidine is 1.4. In one embodiment, the molar ratio of compound of formula (1) and L- histidine is 1.5. In another embodiment, the molar ratio of compound of formula (1) and L- histidine is 1.6. In another embodiment, the molar ratio of compound of formula (1) and L- histidine is 1.7. In another embodiment, the molar ratio of compound of formula (1) and L- histidine is 1.71. In another embodiment, the molar ratio of compound of formula (1) and L- histidine is 1.72. In another embodiment, the molar ratio of compound of formula (1) and L- histidine is 1.73. In another embodiment, the molar ratio of compound of formula (1) and L- histidine is 1.74. In another embodiment, the molar ratio of compound of formula (1) and L- histidine is 1.75. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is 1.8. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is 1.9. In another embodiment, the molar ratio of compound of formula (1) and L-histidine is 2.

[0049] In some embodiments, the pharmaceutical composition is an aqueous composition, wherein compound of formula (1) is in a concentration in the range of Img / ml to 50 mg / ml. In one embodiment, compound of formula (1) is in a concentration in the range of 5 mg / ml to 45 mg / ml. In another embodiment, compound of formula (1) is in a concentration in the range of 10 mg / ml to 40 mg / ml. In another embodiment, compound of formula (1) is in a concentration in the range of 15 mg / ml to 35 mg / ml. In another embodiment, compound of formula (1) is in a concentration in the range of 20 mg / ml to 30 mg / ml.

[0050] In one embodiment, the pharmaceutical composition is an aqueous composition, wherein compound of formula (1) is in a concentration of 1 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 5 mg / ml. In another embodiment,compound of formula (1) is in a concentration of 10 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 15 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 20 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 25 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 30 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 35 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 40 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 45 mg / ml. In another embodiment, compound of formula (1) is in a concentration of 50 mg / ml.

[0051] In one embodiment, the pharmaceutical composition comprises 2 % w / v compound of formula (1). In another embodiment, the composition forther comprises HPMC , glycerol, L-histidine, NaCl and WFI.

[0052] In one embodiment, the pharmaceutical composition comprises 2 % w / v (as calculated as free base) of the solvate salt of compound of formula (1). In another embodiment, the composition forther comprises HPMC, glycerol, L-histidine, NaCl andWFI.

[0053] In one embodiment, the pharmaceutical composition comprises 2 % w / v (as calculated as free base) of the solvate salt of compound of formula (1) , 0.5 % w / v HPMC , 0.5 % w / v glycerol, 0.62 % w / v L-histidine and WFI ad 100%.

[0054] In one embodiment, the pharmaceutical composition comprises 2.56 % w / v the sodium iso-propanol solvate salt of compound of formula (1) , 0.5 % w / v HPMC , 0.5 % w / v glycerol, 0.62 % w / v L-histidine and WFI ad 100%.Administration

[0055] In some embodiments, the pharmaceutical composition is suitable for ocular administration.

[0056] In some embodiments, the ocular administration is by eye drops, eye creams, eye sprays, intraocular depot formulations, by injection including intraocular injection, intra- or periocular injections, comprising intraocular fluids, intravitreal injections, injections into the supra-arachnoidal or the subretinal space, depot formulations, solid and semisolid intraocular carriers and matrices, or ocular devices.

[0057] In one embodiment, the ocular administration is by eye drops. In another embodiment, the ocular administration is by eye creams. In another embodiment, the ocular administration is by eye sprays. In another embodiment, the ocular administration is by intraocular depot formulations. In another embodiment, the ocular administration is by injection including intraocular injection, intra- or periocular injections, comprising intraocular fluids, intravitreal injections, injections into the supra-arachnoidal or the subretinal space. In another embodiment, the ocular administration is by depot formulations. In another embodiment, the ocular administration is by solid and semisolid intraocular carriers and matrices. In another embodiment, the ocular administration is by ocular devices.Medical Use

[0058] In some embodiments, the pharmaceutical composition is for the treatment of an ophthalmic disorder or disease.

[0059] In some embodiments, the ophthalmic disease or disorder comprises amyloid- associated retinal disease.

[0060] The phrase "amyloid-associated disease or disorder”, as used herein, describes a medical condition with a pathology that involves amyloid beta formation, specifically thetoxic oligomeric species of amyloid beta. Such a medical condition can involve other pathologies, yet, is treatable, at least to some extent, by reducing, preventing or inhibiting amyloid toxicity and / or amyloid plaque formation.

[0061] In some embodiments, the amyloid-associated retinal disease is selected from the group consisting of primary angle-closure glaucoma, secondary open-angle glaucoma, wide-angle glaucoma, steroid-induced glaucoma, traumatic glaucoma, secondary angleclosure glaucoma, neovascular glaucoma, pigmentary dispersion syndrome, pseudoexfoliation syndrome, uveitis and glaucoma and diabetic retinopathy.

[0062] In one embodiment, the amyloid-associated retinal disease is primary angleclosure glaucoma. In another embodiment, the amyloid-associated retinal disease is secondary open-angle glaucoma. In another embodiment, the amyloid-associated retinal disease is wide-angle glaucoma. In another embodiment, the amyloid-associated retinal disease is steroid-induced glaucoma. In another embodiment, the amyloid-associated retinal disease is traumatic glaucoma. In another embodiment, the amyloid-associated retinal disease is secondary angle-closure glaucoma. In another embodiment, the amyloid- associated retinal disease is neovascular glaucoma. In another embodiment, the amyloid- associated retinal disease is pigmentary dispersion syndrome. In another embodiment, the amyloid-associated retinal disease is pseudo-exfoliation syndrome. In another embodiment, the amyloid-associated retinal disease is uveitis and glaucoma. In another embodiment, the amyloid-associated retinal disease is diabetic retinopathy.

[0063] In one embodiment, the amyloid-associated retinal condition or condition of the optic nerve is related to any mechanical trauma of the eye leading to macroscopic or microscopic injuries or damages.

[0064] The exact dose and regimen of administration of the pharmaceutical composition will necessarily be dependent upon the therapeutic or nutritional effect to be achieved and may vary with the particular formula, the route of administration, and the age and condition of the individual subject to whom the pharmaceutical composition is to be administered.

[0065] Suitable dosage ranges for eye drops are 0.1 to 50 milligrams daily, preferably 0.5 to 20 milligrams daily, and especially 0.5 to 10 milligrams daily, depending upon the exact mode of administration, form in which administered, the indication toward which the administration is directed, the subject involved, and the body weight of the subject involved, and the preference and experience of the physician or veterinarian in charge. In one embodiment, the term "therapeutically effective" applied to dose or amount refers to that quantity of a compound or pharmaceutical composition that is sufficient to result in a desired activity upon administration to a living animal body in need thereof.

[0066] Suitable dosage ranges for administrations to the eye are in the range of 0. Img to 20 mg per single topical application. In other embodiments, suitable dosage ranges for administrations to the eye are in the range of 0.5 mg to 15 mg per single topical application. In other embodiments, suitable dosage ranges for administrations to the eye are in the range of 1 mg to 10 mg per single topical application. In other embodiments, Suitable dosage ranges for administrations to the eye are in the range of 2 mg to 6 mg per single topical application. In some embodiment, the applications into the eye are administered once a day, twice a day or three times a day. In some embodiments, the applications into the eye are administered to a maximum of 9 times per day.

[0067] In some embodiments, the pharmaceutical composition is administered as an initial loading dose and further in multiple subsequent maintenance doses.

[0068] In some embodiments, the initial dose is administered 1-3 times per day for one day. In other embodiments, the initial dose is given 1-3 times per day for two days. In other embodiments, the initial dose is given 1-3 times per day for three days. In other embodiments, the initial dose is given 1-3 times per day for one week. In other embodiments, the initial dose is given 1-3 times per day for two weeks. In other embodiments, the initial dose is given 1-3 times per day for three weeks. In other embodiments, the initial dose is given 1-3 times per day for one month.

[0069] In some embodiments, the total dose is administered at one time point as several single drops given in short time intervals that allow the absorption of the dose before the next dose is given. Such time intervals can be e.g. 30 sec up to 15 min, typically in 5 minute intervals. For example, administration is 3 times a day with 3 drops at each administration time point given in 5 min intervals. That results in administration of 9 single doses.

[0070] In some embodiments, the maintenance dose is administered once a week. In other embodiments, the maintenance dose is administered twice a week. In other embodiments, the maintenance dose is administered several times a week. In other embodiments, the maintenance dose is administered once every two weeks. In other embodiments, the maintenance dose is administered once every three weeks. In other embodiments, the maintenance dose is administered several times per fortnight. In other embodiments, the maintenance dose is administered once a month. In other embodiments, the maintenance dose is administered several times a month. In other embodiments, the maintenance dose is administered once every two months. In other embodiments, the maintenance dose is administered once every three months.

[0071] In some embodiments, the maintenance dose is administered for one month. In other embodiments, the maintenance dose is administered for two months. In otherembodiments, the maintenance dose is administered for three months. In other embodiments, the maintenance dose is administered for six months. In other embodiments, the maintenance dose is administered for one year. In other embodiments, the maintenance dose is administered for more than one year.

[0072] In some embodiments, the maintenance dose is 10%-75% of the initial dose. In other embodiments, the maintenance dose is 20%-75% of the initial dose. In other embodiments, the maintenance dose is 25%-75% of the initial dose.

[0073] The term “treatment” as used herein refers to the administering of a therapeutic amount of the pharmaceutical composition of the present invention which is effective to ameliorate undesired symptoms associated with a disease, to prevent the manifestation of such symptoms before they occur, to slow down the progression of the disease, slow down the deterioration of symptoms, to enhance the onset of remission period, slow down the irreversible damage caused in the progressive chronic stage of the disease, to delay the onset of said progressive stage, to lessen the severity or cure the disease, to improve survival rate or more rapid recovery, or to prevent the disease form occurring or a combination of two or more of the above.

[0074] The "effective amount" for purposes disclosed herein is determined by such considerations as may be known in the art. The amount must be effective to achieve the desired therapeutic effect as described above, depending, inter alia, on the type and severity of the disease to be treated and the treatment regime. The effective amount is typically determined in appropriately designed clinical trials (dose range studies) and the person versed in the art will know how to properly conduct such trials in order to determine the effective amount. As generally known, an effective amount depends on a variety of factors including the affinity of the ligand to the receptor, its distribution profile within the body, avariety of pharmacological parameters such as half-life in the body, on undesired side effects, if any, on factors such as age and gender, etc.Process

[0075] In some embodiments, disclosed herein is a process for preparation of a pharmaceutical composition, comprising compound of formula (1)and L-histidine, comprising the steps comprising the steps of dissolving, compounding, filtering and filling the aqueous solutions into primary packaging materials.ExamplesExample 1 - Solubility of GAL-101 in the presence of Histidine.

[0076] The aim of this example was to determine solubility of GAL-101 in the presence of L-histidine (His) in deionized water.

[0077] GAL-101 free base has only limited solubility of about 8 mg / ml in water. To achieve higher concentrations for beneficial therapeutic effect, a GAL-101 salt can be used, which has much higher solubility in water than the concurrent free base.

[0078] However, in solution when adding excipients and adjusting the pH a salt frequently behaves similar to the free base after equilibrium has been reached. To generate pharmaceutical formulations with high concentrations, where the active pharmaceutical ingredient does not precipitate over time, solubilizers are frequently applied. The effect of solubilizers was also tested for formulations of GAL-101 salt. Indeed, as can be seen in Fig.1, different solubilizers (cyclodextrines, Kolliphor EL, Polysorbate 80 and TPGS1000) had a beneficial effect on the solubility of GAL-101. Nevertheless, solubilizers are frequently not preferred since they can be irritating to the eye or have other side effects. Therefore, a formulation without solubilizers should be developed. Surprisingly, it was found that the amino acid L-Histidine, which can be used as a buffer substance, has also a beneficial effect on solubility of GAL-101 salt which other typically used buffer materials do not have. Data is illustrated in Fig. 2. As can be seen in surprisingly only histidine has a significant effect on solubility of GAL-101.

[0079] The inventors further tested the stability of the compositon and determined that based on the 12 months stability data obtained so far for the commercial product a shelflife of 36 months seems justified.

[0080] EXAMPLE 2 - Ocular Pharmacokinetic Study Following a Single Ocular Topical Administration of Gal-101 formulation in Pigmented Rabbits.

[0081] The aim of this study was to investigate the ocular pharmacokinetic profile of GAL-101 salt as active substance in a formulation comprising L — Histidine following a single ocular topical administration in pigmented rabbits.

[0082] The GAL-101 formulation consisted of GAL-101 solvent salt 2.56% w / v (corresponding to GAL-101 free base 2.0% w / v), HPMC 0.5% w / v, glycerol 0.5% w / v, L- histidine 0.5% w / v, citric acid q.s pH 6.5, water for injection to 1.0 mL.

[0083] Test and control (placebo) items were instilled in both eyes of each rabbit (30 pL in a single administration) using an appropriate micropipette. Each day, the general clinical signs and the appearance of all animals was observed.

[0084] The study groups are described in Table :

[0085] Table 2: Study GroupsThe study schedule is described in Table 3:Table 3: Study ScheduleAbbreviations: C=Comea; CH=Choroid; CJ=Conjunctivae; ICB=Iris Ciliary Body; PL=Plasma; R=Retina; V= Vitreous; WB=Whole Blood.

[0086] Both eyes of each rabbit were examined using an ophthalmoscope which allowed a precise observation of the conjunctivae, cornea and iris reactions. The observations were scored using the Draize’s scale [Draize JH, Woodgard G and Calvery HO. Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes. J. Pharmacol. Exp. Ther. 1944; 82: 377-390],

[0087] Animals were anesthetized for blood sampling. At the end of the measurement period, animals were euthanized by an intracardiac injection of overdosed pentobarbital following an anaesthesia. Immediately after euthanasia, conjunctivae (pooled bulbar & palpebral), cornea, iris ciliary body, retina, choroid and vitreous were quickly and carefullydissected from both eyes, put in tubes, weighed, vortexed and temporarily stored in dry ice, then at -23°C ± 7°C until assay. The analysis for the content of GAL-101 in PL or ocular samples was performed by RRLC-MS / MS method.

[0088] Preliminary results indicate a very good local and systemic tolerability of the tested formulation.

[0089] Preliminary results indicate GAL-101 levels in the cornea, choroid, conjunctivae, iris, ciliary body, retina, vitreous and plasma for the tested formulation exceeding levels necessary for pharmacological activity.EXAMPLE 3 - GAL-101 - 28-day evaluation of the ocular tolerance and systemic toxicokinetic following repeated ocular topical administrations of GAL-101 formulation in pigmented rabbits, followed by 1-week recovery period

[0090] The aim of this study was to evaluate the ocular tolerance of GAL-101 and the systemic exposure to GAL-101 by measuring the test item concentration in plasma following repeated ocular topical administrations of GAL-101 formulation for 1 month, followed by 1-week recovery period, in pigmented rabbits.

[0091] 32 animals (16 males and 16 females) were included in this study. GAL-101 formulation or vehicle was instilled in the right eyes (1 or 4 drops of 30 pL per administration).

[0092] The in-life part of the study is described in Table 4:

[0093] Table 4: Study Design 28-day ocular tolerance study in rabbits.

[0094] The study schedule is described in Table :

[0095] Table 5: Study Schedule 28-day ocular tolerance study in rabbits

[0096] Ocular examinations of both eyes included ophthalmoscopy using the Draize’s scale, slit-lamp examination, intraocular pressure (IOP) measurement, full-field electroretinographic responses,• comeal sensitivity of both eyes will be tested using an esthesiometer and• comeal thickness evaluation with pachymetry

[0097] Blood was collected for determination of GAL-101 plasma levels by RRLC- MS / MS method.

[0098] Immediately after euthanasia, both eyeballs (including optic nerve, conjunctivae, and extraocular muscles), with Harderian and orbital lacrimal glands, nictitating membrane and both eyelids were sampled, fixed, and processed for microscopic examination.

[0099] Preliminary results indicate a very good local and systemic tolerability as well as no adverse histopathological findings.[000100] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims

CLAIMSWhat is claimed is:

1. A pharmaceutical composition comprising compound of formula (1)and L-histidine.

2. A pharmaceutical composition according to claim 1, comprising a salt of compound of formula (1).

3. A pharmaceutical composition according to claims 1-2, comprising a solvate salt compound of formula (1), wherein said solvate is a C3 orC4 alcohol thereof.

4. A pharmaceutical composition according to claim 3, wherein said solvate salt compound of formula (1) is a crystalline solvate salt.

5. A pharmaceutical composition according to claims 3-4, wherein said salt is an alkali salt.

6. A pharmaceutical composition according to claim 5, wherein said alkali salt is selected from sodium and potassium.

7. A pharmaceutical composition according to claims 3-6, wherein solvate is selected from 1 -propanol, 2-propanol, 1 -butanol, iso-butanol and any combinations thereof.

8. A pharmaceutical composition according to claims 1-7, further comprising citric acid, citrate or combination thereof.

9. A pharmaceutical composition according to claims 1-8, wherein said composition has pH in the range of 5-8.

10. A pharmaceutical composition according to claim 9, wherein said composition has a pH in the range of 6.0-7.0.

11. A pharmaceutical composition according to claims 1-10, wherein the molar ratio of compound of formula (1 ) and L-histidine is in the range 1.4-2.0.

12. A pharmaceutical composition according to claim 11, wherein the molar ratio of compound of formula (1 ) and L-histidine is in the range 1.70-1.73.

13. A pharmaceutical composition according to claims 1-12, wherein said composition is an aqueous composition, wherein compound of formula (1 ) is in a concentration in the range of Img / ml to 50 mg / ml.

14. A pharmaceutical composition according to claim 13, wherein compound of formula (1 ) is in a concentration of 20 mg / ml.

15. A pharmaceutical composition according to claims 1-14, suitable for ocular administration.

16. A pharmaceutical composition according to claim 15, wherein said ocular administration is by eye drops, eye creams, eye sprays, intraocular depot formulations, by injection including intraocular injection, intra- or periocular injections, comprising intraocular fluids, intravitreal injections, injections into the supra-arachnoidal or the subretinal space, depot formulations, solid and semisolid intraocular carriers and matrices, or ocular devices.

17. A pharmaceutical composition according to claim 15, wherein said ocular administration is by eye drops.

18. A pharmaceutical composition according to claims 1-17, for the treatment of an ophthalmic disorder or disease.

19. A pharmaceutical composition for use according to claim 18, wherein said ophthalmic disease or disorder comprises amyloid-associated retinal disease.

Images