Novel Solution

Abstract

The present invention provides a pharmaceutical composition, which is an aqueous solution containing Compound (I) (also known as U0126) or a pharmaceutically acceptable salt thereof at a concentration of 50-200 μM, further comprising 0.2-1.0% (v / v) polyoxyethylene castor oil. The composition is particularly useful in treating ischemic injury (particularly subarachnoid hemorrhage) in human patients by direct injection into the brain. Uses of the composition, unit doses containing the composition, and methods of using the composition are also disclosed. [Formula 1] TIFF2026501344000012.tif40159

Description

[Technical Field]

[0001] The present invention relates to solutions of pharmaceutically active compounds, particularly for use in the treatment of ischemic injury, particularly in the treatment of ischemic injury in the brain. Specifically, the present invention relates to an aqueous solution comprising a pharmaceutically acceptable solvate of the compound known as U0126 or a salt thereof, and its use in the treatment of ischemic injury. [Background technology]

[0002] Subarachnoid hemorrhage (SAH) is an uncommon form of stroke caused by bleeding in the space surrounding the brain. Most often, it occurs when a weak area (aneurysm) in a blood vessel on the surface of the brain ruptures and leaks. Blood then accumulates around the brain and inside the skull, increasing pressure on the brain. SAH is difficult to treat and can be fatal.

[0003] One of the major complications of SAH is secondary cerebral ischemia (also called delayed ischemia) caused by vasospasm, narrowing of blood vessels and restriction of cerebral blood flow. Blood supply to parts of the brain can be dangerously reduced, thereby interfering with normal brain function.

[0004] After confirming that SAH is caused by a cerebral aneurysm, surgical treatment may be recommended to repair the affected blood vessel and prevent the aneurysm from rupturing again. This can be performed using one of two main techniques: coiling (embolization) and clipping. Clipping involves opening the skull, locating the aneurysm, and placing a clip around the neck of the aneurysm. Coiling involves advancing a catheter through an artery supplying the brain from the groin and inserting a platinum coil inside the aneurysm, forming a clot that restricts blood flow.

[0005] Secondary cerebral ischemia can be alleviated by pharmacological intervention with the calcium channel blocker nimodipine. The recommended dose of nimodipine by the UK National Institute for Health and Care Excellence is 60 mg every 4 hours, starting within 4 days of aneurysmal subarachnoid hemorrhage and continuing for 21 days.

[0006] Preliminary preclinical studies have shown that compound U0126 shows promise for the treatment of SAH. U0126 has the formula: [ka] It has.

[0007] It is known to be an inhibitor of MEK1 and MEK2 and has been proposed for use in the treatment of ischemic disorders, such as subarachnoid hemorrhage. Duncia et al., Bioorg. & Med. Chem. Lett. 8 (1998) 2839-2844, describes this activity. European Patent No. 1,139,512 discloses the use of U0126 for administration from 1 to 6 hours after the onset of ischemic injury. J. Cerebral Blood Flow & Metabolism, 35 (2015) 454-460 discloses that U0126 improves long-term neurological outcomes after stroke in female rats.

[0008] U0126 is a difficult compound to formulate and administer; it is highly insoluble in most common solvents and sensitive to both acids and bases. Formulations that are soluble in physiological fluids and can therefore deliver the active substance directly to the brain for treatment of ischemic disorders are needed for the treatment of ischemic disorders. Intracranial injection is one necessary administration method, which means that the volume of liquid used as a carrier must be limited. Furthermore, the stability of liquid formulations can be problematic. Although U0126 is known to be soluble in DMSO, this solvent is not suitable for use in liquid formulations, particularly those intended for intracranial injection.

[0009] Christensen et al., Drug Delivery, 2019, 26(1), 680-688, describes the composition of U0126 in a 1% solution of Kolliphor EL (also known as Cremophor). Kolliphor EL is a type of polyethoxylated castor oil prepared by reacting 35 moles of ethylene oxide with 1 mole of castor oil.

[0010] In Christensen et al., ex vivo and in vivo experiments were performed to determine the optimal ex vivo dose of U0126, and the toxicity of this dose was also tested in vivo to determine the synergistic effect between Cremaphor and U0126. The toxicological effects of different concentrations of U0126 in rats were investigated. Animals were exposed to three different concentration levels: a low concentration (1x10 -6 ), medium concentration (1×10 -5 M) and high concentrations (2 × 10 -5 M) of U0126 was tested. 15 μL was injected in each case. Neurological observations showed that low and medium doses of U0126 were well tolerated. However, at higher levels, there were signs of neurological impairment. Based on this, the authors performed a maximum of 10 doses in 0.5% Cremophor. -5 We conclude that the M concentration of U0126 (i.e., 10 μM with 0.5% Cremophor in aqueous solution) is the maximal tolerated in vivo dose for intracerebroventricular (ICV) administration.

[0011] There remains a need for improved treatments for ischemic injury, particularly in the brain. Summary of the Invention

[0012] The present invention relates to compound (I): [ka] or a pharmaceutically acceptable salt thereof at a concentration of 50 to 200 μM, The pharmaceutical composition is provided, wherein the solution further contains 0.2 to 1.0% (v / v) polyoxyethylene castor oil.

[0013] The present invention also provides a method for the treatment of ischemic injury, which comprises administering to a human patient an aqueous solution of Compound (I) as described above.

[0014] The present invention also provides an aqueous solution of Compound (I) as described above for the manufacture of a medicament for the treatment of ischemic injury.

[0015] The present invention also provides a unit dose comprising the above aqueous solution of Compound (I), the unit dose having a volume of 1 to 10 mL. DETAILED DESCRIPTION OF THE INVENTION

[0016] As mentioned above, the present invention provides compound (I): [ka] or a pharmaceutically acceptable salt thereof at a concentration of 15 to 200 μM. The compound of formula (I) is known as U0126. Its chemical name is 1,4-diamino-2,3-dicyano-1,4-bis(o-aminophenylmercapto)butadiene.

[0017] The present invention also provides an aqueous solution containing Compound (I) or a pharmaceutically acceptable salt thereof at a concentration of 15 to 200 μM.

[0018] The U0126 used in the pharmaceutical composition or solution of the present invention may be in the form of free U0126 or one of its pharmaceutically acceptable salts, such as a salt with a mineral acid or a salt with any other acid capable of forming a salt with U0126.

[0019] U0126 forms solvates, which can be used to prepare compositions or solutions of the present invention. Suitable solvates include alcohols, e.g., C 1-4 Solvates with alkanols, such as ethanol, are included. Hemiethanolate, ethanolate and diethanolate forms all exist and may be used.

[0020] The inventors surprisingly found that the solution was tolerated and stable in the body at concentrations of 15-200 μM, which was not expected to be tolerated in the body based on previous reports that 10 μM was the maximally tolerated dose.

[0021] In a preferred embodiment, the solution contains Compound (I) at a concentration of about 25 to about 150 μM, for example, about 25 to about 100 μM. More preferably, the concentration is about 50 to about 90 μM, for example, about 70 to about 80 μM. For example, the solution of the present invention contains Compound (I) at a concentration of about 75 μM.

[0022] In terms of the molar mass of U0126 of 380.5 g / mol, the solution contains compound (I) at a concentration of about 9.5 to about 76.1 μg / mL. More preferably, the concentration is about 9.5 to about 57.1 μg / mL, for example, about 9.5 to about 38.5 μg / mL. More preferably, the concentration is about 19.0 to about 34.2 μg / mL, for example, about 26.6 to about 30.44 μg / mL. For example, the solution of the present invention contains compound (I) at a concentration of about 28.5 μg / mL.

[0023] The term "about" refers to a tolerance range of ±20% of the relevant value, such as ±15% of the relevant value, such as ±10% of the relevant value or ±5% of the relevant value, such as ±2% of the relevant value.

[0024] The pharmaceutical compositions of the present invention offer the highly beneficial advantage that they can be used in sufficiently small amounts (typically 0.1 to 5 mL, preferably less than 2 mL) within the brain so as not to be harmful to the brain, yet still provide a sufficient amount of drug to be effective.

[0025] When administered in a volume of 0.2 to 5 mL, the pharmaceutical composition of the present invention provides a patient with a dose of 0.003 to 1.0 μmol of U0126. Since U0126 has a molar mass of 380.5 g / mol, this corresponds to a dose of 1.14 to 380.5 μg of U0126. Preferably, the volume and concentration are selected to provide a dose of 5 to 100 μg of U0126, preferably 5 to 80 μg, e.g., 5 to 50 μg. In a more preferred embodiment, the dose is in the range of 10 to 40 μg. Even more preferably, the dose is in the range of 20 to 30 μg. For example, if a patient is given a dose of 0.33 mL of a 75 μM U0126 solution, it is a dose of 8.55 μg. If a patient is given a dose of 1.0 mL of a 75 μM U0126 solution, it is a dose of 28.5 μg.

[0026] As described below, a course of treatment may involve multiple doses of a drug being given to a patient.

[0027] In a preferred embodiment of the present invention, the solution further contains polyoxyethylene castor oil. Castor oil polyoxyethylates are condensation products of ethylene oxide and castor oil, and several are commercially available. Such polyoxyethylene castor oil derivatives can be prepared, for example, by reacting castor oil with ethylene oxide in a molar ratio of, for example, 1:30 to 1:40, e.g., 1:35. Such products are called macrogolylycerol ricinoleate, PEG-35 castor oil, polyoxyl 35 hydrogenated castor oil, and polyoxyl-35 castor oil. For example, polyoxyl 35 castor oil is available under the trade name Kolliphor® EL, formerly known as "Cremophor EL" (trademark).

[0028] The pharmaceutical composition or solution of the present invention may contain polyoxyethylene castor oil at a concentration of 0.2 to 1.0% (v / v). For example, the solution may contain polyoxyethylene castor oil at a concentration of about 0.3 to about 0.7% (v / v), e.g., about 0.5% (v / v). For example, it may contain polyoxyl 35 castor oil at a concentration of about 0.3 to about 0.7% (v / v), e.g., about 0.5% (v / v).

[0029] The term "solution" is used herein to describe a homogeneous mixture of a solvent (e.g., water) and a solute (particularly U0126). The mixture passes through a filter (e.g., a 0.20 μm filter) without any of the composition being retained in the filter. In some circumstances, a solution, when expanded sufficiently, can be a suspension in which a first solute is surrounded by a second solute, with the second solute in contact with the bulk solvent.

[0030] The solid form of compound U0126 remains intact under ambient conditions for decades. The solutions described herein have been found to remain intact for several hours to an extent acceptable for pharmaceutical use. A solution that remains intact for an amount of time required from preparation to administration to a patient (e.g., 2 hours, e.g., 1 hour) is considered "stable" herein. It has also been found that the solution can be frozen, and that degradation in the solid state occurs only very slowly.

[0031] In certain embodiments, it may be advantageous for the pharmaceutical composition or solution to further comprise a buffer solution.

[0032] In some embodiments, the solution further comprises artificial cerebrospinal fluid (aCSF).

[0033] "Artificial cerebrospinal fluid" is a technical term, and aCSF is widely used in research and medicine, often as an improvement over the use of sterile saline. The composition of aCSF is designed to mirror as closely as possible the composition of human cerebrospinal fluid. Such fluids are sterile aqueous solutions of electrolytes containing sodium, potassium, calcium, and magnesium cations, along with anions selected from phosphate, sulfate, chloride, and bicarbonate. Sugars such as glucose or dextrose may also be included. A typical aCSF contains, for example, Na + : 130mM~170mM, K + : 2mM~5mM, Ca 2+ : 1mM~2.5mM, Mg 2+ : 0.5 mM to 2.5 mM. Preferably, the aCSF contains Na + : 140mM~160mM, K + : 2.5mM~4.5mM, Ca 2+ : 1mM~2mM, Mg 2+ : 0.5 mM to 1.5 mM. More preferably, the aCSF contains Na + : 145mM~155mM, K + : 2.5mM~4mM, Ca 2+ : 1mM~1.5mM, Mg 2+ : 0.5 mM to 1.3 mM. Most preferably, the aCSF contains Na + : Approximately 150mM, K + : 3mM~4mM, Ca 2+ : 1.2mM~1.5mM, Mg 2+ : Contains 0.7mM to 1.3mM.

[0034] Counterions included in aCSF can be, for example, 0.3 mM to 1.5 mM phosphate, 130 mM to 165 mM chloride, and / or 20 mM to 30 mM bicarbonate.

[0035] In certain embodiments, the aCSF is + : 140mM~160mM, K + : 2.5mM~4.5mM, Ca 2+ : 1mM~2mM, Mg 2+: 0.5mM to 1.5mM, P: 0.3mM to 1.5mM, and Cl - : 130 mM to 165 mM. Preferably, the aCSF contains Na + : 145mM~155mM, K + : 2.5mM~4mM, Ca 2+ : 1mM~1.5mM, Mg 2+ : 0.5mM to 1.5mM, P: 0.4mM to 1.2mM, and Cl - More preferably, the aCSF contains 130 mM to 155 mM of Na + : 145mM~155mM, K + : 3mM~4mM, Ca 2+ : 1mM~1.5mM, Mg 2+ : 0.7mM to 1.3mM, P: 0.4mM to 1.1mM, and Cl - : Contains 145mM to 155mM.

[0036] One suitable aCSF is Na + :150mM, K + : 3mM, Ca 2+ : 1.4mM, Mg 2+ : 0.8 mM, P: 1.0 mM, and Cl - Contains :155mM.

[0037] In another embodiment, the aCSF is present in an amount of: 3500-3800 mg of sodium chloride, 900-1000 mg sodium bicarbonate, 350mg to 450mg of dextrose, 120mg-180mg magnesium sulfate·7H2O, 120mg to 180mg of potassium chloride, 80mg-120mg calcium chloride·2H2O, 45 mg to 60 mg of dibasic sodium phosphate anhydrous, and The balance is distilled water.

[0038] In some embodiments, the aCSF is present in an amount of: 3600-3700 mg sodium chloride, 930-990 mg sodium bicarbonate, 375mg to 425mg of dextrose, 140mg-160mg magnesium sulfate·7H2O, 140mg-160mg potassium chloride, 90mg-110mg calcium chloride·2H2O, 50 mg to 55 mg of dibasic sodium phosphate anhydrous, and The balance is distilled water.

[0039] In some embodiments, the aCSF is present in an amount of: 3650-3690 mg sodium chloride, 950-970 mg sodium bicarbonate, 390mg to 410mg of dextrose, 145mg-155mg magnesium sulfate·7H2O, 145mg-155mg potassium chloride, 95mg-105mg of calcium chloride·2H2O, 50 mg to 55 mg of dibasic sodium phosphate anhydrous, and The balance is distilled water.

[0040] A particularly preferred aCSF is: 3670mg sodium chloride, 960 mg sodium bicarbonate, 400mg dextrose, 150 mg magnesium sulfate 7H2O, 150 mg potassium chloride, 100 mg calcium chloride·2H2O, 53 mg of dibasic sodium phosphate anhydrous, and The remainder is distilled water.

[0041] In some embodiments, the aCSF contains, per mL of artificial cerebrospinal fluid: 6.0 to 8.0 mg sodium chloride, 0.2-0.4 mg potassium chloride, 0.15-0.25 mg calcium chloride·2H2O, 0.05 to 0.15 mg of magnesium chloride hexahydrate, 1.2 to 2.3 mg sodium bicarbonate, 0.05 to 0.15 mg disodium phosphate dihydrate, 4.5 mg of Ca as a 1 M solution in hydrochloric acid, and The remainder is distilled water.

[0042] More preferably, aCSF contains: 6.5-7.5 mg sodium chloride, 0.25-0.35 mg potassium chloride, 0.15-0.25 mg calcium chloride·2H2O, 0.05 to 0.15 mg of magnesium chloride hexahydrate, 1.5 to 2.0 mg sodium bicarbonate, 0.05 to 0.15 mg disodium phosphate dihydrate, 4.5 mg of Ca as a 1 M solution in hydrochloric acid, and The remainder is distilled water.

[0043] For example, aCSF contains: 7.2 mg sodium chloride, 0.3 mg potassium chloride, 0.2 mg calcium chloride·2H2O, 0.1 mg magnesium chloride hexahydrate, 1.8 mg sodium bicarbonate, 0.1 mg disodium phosphate dihydrate, 4.5 mg of Ca as a 1 M solution in hydrochloric acid, and The remainder is distilled water.

[0044] Therefore, the present invention provides the following per mL: 9.5–76.1 μg of U0126; 3.0 to 7.0 mg of polyoxyethylene castor oil, 6.0 to 8.0 mg sodium chloride, 0.2-0.4 mg potassium chloride, 0.15-0.25 mg calcium chloride·2H2O, 0.05 to 0.15 mg of magnesium chloride hexahydrate, 1.2 to 2.3 mg sodium bicarbonate, 0.05 to 0.15 mg disodium phosphate dihydrate, 4.5 mg of Ca as a 1 M solution in hydrochloric acid, and A pharmaceutical composition or solution is provided, the balance of which comprises distilled water.

[0045] Preferably, the pharmaceutical composition or solution contains, per mL: 12.5–50.0 μg of U0126; 3.5 to 6.5 mg of polyoxyethylene castor oil, 6.5-7.5 mg sodium chloride, 0.25-0.35 mg potassium chloride, 0.15-0.25 mg calcium chloride·2H2O, 0.05 to 0.15 mg of magnesium chloride hexahydrate, 1.5 to 2.0 mg sodium bicarbonate, 0.05 to 0.15 mg disodium phosphate dihydrate, 4.5 mg of Ca as a 1 M solution in hydrochloric acid, and The balance is distilled water.

[0046] For example, a pharmaceutical composition or solution of the present invention consists essentially of the above ingredients.

[0047] More preferably, the pharmaceutical composition or solution contains, per mL: 19.0–38.0 μg of U0126; 4.0 to 6.0 mg of polyoxyethylene castor oil, 6.5-7.5 mg sodium chloride, 0.25-0.35 mg potassium chloride, 0.175-0.225 mg of calcium chloride·2H2O, 0.075 to 0.125 mg of magnesium chloride hexahydrate, 1.5 to 2.1 mg sodium bicarbonate, 0.075 to 0.125 mg disodium phosphate dihydrate, 4.5 mg of Ca as a 1 M solution in hydrochloric acid, and The balance is distilled water.

[0048] For example, a pharmaceutical composition or solution of the present invention consists essentially of the above ingredients.

[0049] For example, the pharmaceutical composition or solution may contain, per mL: 28.5 μg U0126, 5.0 mg polyoxyethylene castor oil, 6.0 mg sodium chloride, 0.30 mg potassium chloride, 0.20 mg calcium chloride·2H2O, 0.10 mg magnesium chloride hexahydrate, 1.8 mg sodium bicarbonate, 0.10 mg disodium phosphate dihydrate, 4.5 mg of Ca as a 1 M solution in hydrochloric acid, and The balance is distilled water.

[0050] For example, a pharmaceutical composition or solution of the present invention consists essentially of the above ingredients.

[0051] Preferably, the pharmaceutical composition or solution of the present invention is essentially free of dimethyl sulfoxide (DMSO), however, in some embodiments, the pharmaceutical composition or solution may further contain dimethyl sulfoxide (DMSO) in an amount sufficiently low that it is tolerated by the human body.

[0052] The pharmaceutical compositions or solutions according to the present invention may contain one or more additional therapeutic agents. Any desired additional active ingredient may be used. For example, the additional therapeutic agent may be selected from an anti-inflammatory agent (e.g., a steroid, e.g., dexamethasone), a calcium channel blocker (e.g., nimodipine), or an inhibitor of a component of the inflammatory response (e.g., a TNFα or interleukin blocker, typically an antibody).

[0053] The solution can be prepared by a process that includes mixing the ingredients together, followed by filtration, e.g., microfiltration, if necessary. The use of vigorous solubilization techniques, such as vigorous stirring or shaking, or the application of ultrasound, may be desirable. Mixing can be carried out at any suitable temperature and / or pressure. Preferably, the ingredients are mixed together at a temperature between 20°C and 45°C, preferably between 25°C and 40°C, and most preferably between 25°C and 30°C. Furthermore, the ingredients are preferably mixed together at atmospheric pressure (e.g., 1 atmosphere). Because the compositions of the present invention should be sterile, the process of the present invention should be carried out under controlled conditions; for example, the process may include sterile filtration with cooling.

[0054] The solution may be prepared extemporaneously or may be prepared in advance and provided in unit or multi-dose form. It may also be provided in the form of a kit, i.e., in the form of two or more separate components, where component (a) containing U0126 is provided together with component (b) containing polyoxyethylene castor oil and, optionally, component (c) aCSF. Preferably, component (a) is provided as a powder (e.g., a lyophilized powder of U0126). The powder can then be combined with components (b) and (c) to form the solution of the present invention. Thus, the present invention further provides a kit suitable for preparing the solution of the present invention, comprising, as separate components, (a) a powder containing U0126 and (b) polyoxyethylated castor oil. The kit may also optionally contain (c) a pharmaceutically acceptable isotonic liquid (e.g., aCSF). The kit may include a dry component (such as a powder) containing U0126 and a liquid component containing a mixture of polyoxyethylated castor oil and a pharmaceutically acceptable isotonic fluid (e.g., aCSF). The kit may also include a dry component (such as a powder) containing U0126 and a dry mass formulation that comprises a pharmaceutically acceptable isotonic fluid (e.g., aCSF), as well as a liquid component containing polyoxyethylated castor oil.

[0055] The masses of the solid components and volumes of the liquid components in the kits of the invention are selected to provide, when reconstituted, a pharmaceutical composition or solution of the invention. The kits are preferably provided with instructions for the preparation of the pharmaceutical composition or solution of the invention as defined herein.

[0056] For example, immediately prior to use, dried U0126 is reconstituted with a pharmaceutically acceptable isotonic solution (e.g., aCSF) in the presence of polyoxyethylated castor oil, the aCSF being provided with the kit or by the pharmacist or medical professional who performs the reconstitution. In embodiments in which the kit includes a powder comprising U0126 and dry mass formulation components that constitute a pharmaceutically acceptable isotonic solution (e.g., aCSF), immediately prior to use, the powder is reconstituted with (sterile) distilled water, thereby forming a pharmaceutically acceptable isotonic solution (e.g., aCSF) in situ. The polyoxyethylated castor oil component of the kit can be added together with or separately from the distilled water (e.g., before or after the addition of the distilled water).

[0057] Suitable powders can be prepared by known methods, for example, lyophilization. Of course, the compositions of the invention should be sterile, and therefore the components supplied as part of the kit should also be sterile.

[0058] The present invention further provides a pharmaceutically acceptable solution comprising polyoxyethylated castor oil and artificial cerebrospinal fluid. Such a solution is particularly useful in the kit of the present invention. For example, such a solution contains 3.0 to 7.0 mg of polyoxyethylated castor oil and aCSF. For example, such a solution contains, per mL: 3.0 to 7.0 mg of polyoxyethylene castor oil, 6.0 to 8.0 mg sodium chloride, 0.2-0.4 mg potassium chloride, 0.15-0.25 mg calcium chloride·2H2O, 0.05 to 0.15 mg of magnesium chloride hexahydrate, 1.2 to 2.3 mg sodium bicarbonate, 0.05 to 0.15 mg disodium phosphate dihydrate, and Contains 4.5 mg of Ca as a 1 M hydrochloric acid solution. The remainder is water.

[0059] Based on the findings described herein, the solutions of the present invention are suitable for use as pharmaceuticals. The pharmaceutical compositions and solutions described herein are particularly useful for treating ischemic disorders, particularly subarachnoid hemorrhage.

[0060] The present invention provides an aqueous solution containing U0126 or a pharmaceutically acceptable solvate or salt thereof at a concentration of 15 to 100 μg for use as a pharmaceutical. Preferred and exemplary forms of the solution described herein are also provided for use as a pharmaceutical.

[0061] Preferably, the solution is for use in treating ischemic injury in a human patient.

[0062] Prior to the present invention, U0126 had never been injected intracranially. Accordingly, the present invention further provides U0126, or a pharmaceutically acceptable salt or solvate thereof, for use in treating ischemic injury by direct injection into the brain of a solution of U0126, or a pharmaceutically acceptable salt or solvate thereof, and a method for treating ischemic injury, comprising directly injecting into the brain a solution comprising U0126, or a pharmaceutically acceptable salt or solvate thereof.

[0063] The present invention also provides a method for treating ischemic injury in a human patient, comprising administering a therapeutically effective amount of a solution of the present invention. Preferably, the composition is administered by injection. In a particularly preferred embodiment, the composition is administered by intracerebroventricular (ICV) injection.

[0064] The present invention also provides the use of the solution of the present invention for the manufacture of a medicament for the treatment of an ischemic disorder, preferably a subarachnoid hemorrhage.

[0065] The solutions of the present invention may be provided in unit dosage form. In one embodiment, the present invention provides a unit containing a volume of 0.2 to 10 mL of solution capable of administering a therapeutic dose. In a preferred embodiment, the unit has a volume of 3 to 9 mL, for example, 5 to 8 mL, e.g., 7 mL. Typically, the dosing volume administered to a patient is in the range of 0.1 to 5 mL. Preferably, it is less than 2 mL, for example, it may be 0.2 to 1.5 mL, preferably 0.3 to 1.0 mL, e.g., 0.33 mL or 1.0 mL. [Example]

[0066] Example 1: Preparation of U0126 solution in 0.5% Cremophor EL (Tocris Pharma) in aCSF A solution of sodium chloride, potassium chloride, calcium chloride dihydrate, and magnesium chloride hexahydrate was prepared by mixing in water (water for injection). The pH was adjusted to 2.8 using 1 M HCl and / or 0.5 M NaOH. A second solution was prepared by mixing sodium bicarbonate and disodium phosphate dihydrate in water (water for injection). The first and second solutions were mixed together to form an aCSF solution.

[0067] The U0126 drug product was dissolved in an aqueous solution of Kolliphor EL. This was then added to the aCSF solution. The solution was prepared in three 1 liter batches, combined to give a final volume of 3 L.

[0068] The solution was sterile filtered using two Millipak 100 0.22 μm PVDF filter units. The solution was then filled into vials, which were then sealed. The product was checked for purity and the correct final concentration. The final concentrations of each component were as shown in Table 1. [Table 1]

[0069] Briefly, the solution contained 28.55 μg / mL of U0126 (75 μM) and 0.5% v / v of Kolliphor EL.

[0070] Example 2: Alternative preparation of a solution of the present invention 0.79 g of Cremophor EL™ was weighed into an Erlenmeyer flask and 60 mL of aCSF (obtained from Tocris Pharma) was added. The mixture was shaken vigorously for 15 minutes to obtain a clear solution containing 1.2% v / v Cremophor EL. 4.88 mg of U0126 was added to the solution along with an additional 15 mL of aCSF. These amounts were calculated to yield 194 μM U0126 in a 1.0% v / v solution of Cremophor EL in aCSF. The solution was then placed in an ultrasonic bath maintained at 25-28°C for 20 minutes. After sonication, some particles were observed in the solution. The solution was sonicated for an additional 25 minutes. The resulting mixture was diluted with an additional 74.25 mL of aCSF solution to yield a calculated 78.75 μM U0126 in a 0.5% v / v solution of Cremophor EL in aCSF. The actual measured content of U0126 was 67 μM (HPLC, Agilent 1100).

[0071] The mixture was then filtered using a Millipore Millex-LG, PTFE, 0.20 μm filter (SLLG025SS), which removed all solid material, leaving a clear solution containing a measured content of 68 μM U0126.

[0072] Example 3: Alternative preparation of a solution of the present invention 500 μL of Cremophor EL was pipetted into an Erlenmeyer flask containing 40 mL of in-house prepared aCSF. The mixture was vigorously shaken for 15 minutes, resulting in a clear solution containing 1.25% Cremophor EL. To this solution, 4.21 mg of U0126 and an additional 10 mL of aCSF were added. The mixture was vigorously shaken for 43 minutes, resulting in a non-clear solution. Shaking for an additional 87 minutes did not dissolve any remaining particles. The mixture was then sonicated for 5 minutes, resulting in particle dissolution. An additional 10 minutes of sonication ensured complete dissolution. The resulting solution was calculated to contain 1.0% Cremophor EL and 200 μM U0126.

[0073] This solution was then diluted with 50 mL of aCSF to yield a solution calculated to contain 0.5% Cremophor EL and 98.7 μM U0126. Various volumes of this solution, with a measured pH of 8.6, were filtered using a Millipore Sterile Syringe Filter, Millex-LG PTFE 0.20 μm (SLLG013SL). Analysis of various samples showed U0126 contents ranging from 85 to 99 μM.

[0074] Biological Example 1: Study in Human Subjects with SAH A randomized, double-blind, placebo-controlled study is conducted to determine the safety and tolerability of the drug product of Example 1 (Investigational Medicinal Product - IMP) administered as one and three ICV bolus injections over an 18-hour period to patients in the early post-subarachnoid hemorrhage (SAH) stage. The study will include five patient cohorts who will receive the drug product of Example 1 (Investigational Medicinal Product - IMP) as follows: Cohort 1: 3 patients, low-dose IMP, no placebo, single dose Cohort 2: 4 patients, medium dose, randomized 3:1 to IMP:placebo, single dose Cohort 3: 4 patients, full dose, randomized 3:1 to IMP:placebo, single dose Cohort 4: 4 patients, full dose, randomized 3:1 to IMP:placebo, multiple doses Cohort 5: 12 patients, full dose, randomized 3:1 to IMP:placebo, multiple doses

[0075] The investigational medicinal product (IMP) is the solution described in Example 1. It contains the active pharmaceutical ingredient U0126 at a concentration of 28.5 μg / mL. Low-dose patients receive 0.33 mL of solution. Medium-dose patients receive 0.6 mL of solution. High-dose patients receive 1.0 mL of solution. Multiple-dose patients receive three doses of 1.0 mL per dose.

[0076] The placebo was 0.9% saline.

[0077] This study has the following inclusion and exclusion criteria:

[0078] Selection criteria: Patients who meet the following criteria are eligible to participate: 1. Male or female patients aged 18 to 80 years (inclusive). 2. Moderate or severe SAH caused (or suspected to be caused) by a ruptured saccular aneurysm diagnosed by CT and with symptoms lasting less than 8 hours. 3. Intraventricular access performed within 8 hours of symptom onset. 4. WFNS score of 1-5 assessed at any time after SAH diagnosis but before the first dose of IMP. 5. Informed consent was obtained from the study guardian prior to the start of any study-related procedures.

[0079] Exclusion criteria: Patients will be excluded from participation in the study if any of the following criteria apply: 1. SAH due to other causes (e.g., trauma, rupture of a fusiform or mycotic aneurysm). 2. Intraventricular or intracerebral blood in the absence of subarachnoid blood. 3. Expected survival time less than 48 hours. 4. Any severe or unstable chronic or acute concomitant condition that the PI / subinvestigator determines may affect the evaluation of the safety of the IMP. 5. Any known or CT evidence of significant previous brain injury or pre-existing cerebrovascular disease that the investigator / subinvestigator believes may affect the accurate diagnosis and evaluation of SAH. 6. Participation in any other clinical trial with an experimental drug within the last 12 weeks. 7. Known allergy to IMP or any of its components. 8. Pregnant or lactating female patients. Women of childbearing potential must have a negative plasma or urine pregnancy test at screening.

[0080] Investigational Medicinal Product (IMP) Preparation and Administration: The IMP will be prepared at the investigational site by an unblinded pharmacist as detailed in the pharmaceutical preparation.

[0081] Administration of IMP or placebo will be by intracerebroventricular injection according to the following schedule, shown in Table 2: [Table 2]

[0082] After administration of 0.33 mL, 0.6 mL, or 1.0 mL of IMP, a 2-5 mL injection of isotonic NaCl is also administered to ensure the entire dose of IMP reaches the target. The ICV connection should remain closed for 15-45 minutes after injection. During this time, intracranial pressure (ICP) should be monitored frequently at the investigator's discretion to avoid excessive ICP buildup.

[0083] For each patient, the following information will be recorded as part of the study protocol: Population statistics (age, gender, height, weight) Medical history and SAH diagnosis Concomitant medications Vital signs Electrocardiogram: Continuous monitoring via 3-lead ECG throughout the entire NICU stay according to clinical practice. Magnetic Resonance Imaging Scan: An MRI scan will be performed at the 84th day visit to document cerebral infarction. World Federation of Neurosurgical Societies Score: The level of consciousness and neurological deficit will be measured by the WFNS score at screening as one of the inclusion criteria and as the baseline status before the first IMP administration. Glasgow Coma Scale (GCS)GCS The GCS is used to assess the patient's level of consciousness during hospitalization. The frequency of administration is at least once daily. The Glasgow Outcome Scale-Extended (GOS-E) is used to assess a patient's health status in eight categories of clinical functioning of health: dead, vegetative state, lower severe disability, upper severe disability, lower moderate disability, upper moderate disability, lower good recovery, and upper good recovery. The modified Rankin Scale (mRS) is used to measure the degree of impairment in daily activities or the degree of dependency on a caregiver. NIHSS: NIHSS grading for focal neurological deficits will be assessed at baseline and as a measure of disease progression up to 14 days after SAH. Laboratory Safety Assessment: Blood samples will be collected for clinical chemistry and hematology analysis. Arterial blood gas parameters will be collected and sent to the local laboratory's certified clinical chemistry laboratory for analysis by routine analytical methods. Blood samples for laboratory safety assessment will be collected prior to each IMP administration (within 1 hour for the second and third doses), 75 minutes (± 15 minutes) after each IMP administration, and then once daily (excluding blood gases unless clinically indicated) for the remainder of the patient's intensive care period in the NICU. Pregnancy test Delayed cerebral ischemia Registration of length of stay in NICU, hospital units, and other hospitals Any exploratory procedures and evaluations that are performed.

[0084] As of the filing of this patent application, Cohorts 1-3 are complete, and Cohort 4 is partially complete. No serious adverse events (SAEs) causally related to the investigational drug product preparation or administration procedures were reported (including Cohort 4, in which patients received multiple doses: three doses of 1.0 mL per dose, as described in the protocol above). Efficacy data readout will be available upon completion of further phases of the study.

[0085] Biological Example 2: Repeated Dose Toxicity Study in Dogs The purpose of this study was to evaluate the toxicity of the drug product of Example 1 (Investigational Medicinal Product - IMP) when administered once daily by ICV injection to beagle dogs for 8 days (days 0-7), with emphasis on effects in the brain. Additionally, cardiovascular and CNS safety pharmacology evaluations were included as an integral part of the study.

[0086] The study consisted of five groups of up to five beagle dogs (maximum three males and two females, see Table 3). A subset of animals underwent magnetic resonance imaging to facilitate localization of the ventricular space. After imaging and analysis of this subset, each animal underwent surgical implantation of an ICV cannula into one of the lateral ventricles. After surgical recovery, Groups 1-5 were administered vehicle and the drug product of Example 1 (Investigational Medicinal Product - IMP) (respectively) via the ICV cannula at three different concentrations in a dose volume of 500 μL per dose (followed by 50 μL of artificial cerebrospinal fluid). The dose levels and the number of animals completing the study are shown in Table 3 below. [Table 3]

[0087] Dosing was by slow ICV injection (approximately 3 μL / sec) once daily for 8 days, for a total of (maximum) 8 doses. Blood for plasma and serum was collected post-dose on days 0 and 7. During the treatment period (days 0-7), animals were monitored for clinical signs during the first hour post-dose and up to four times daily. Body weight and food intake were monitored, and a functional observational battery assessment was performed before dosing began and after dosing on days 0 and 7. Cardiovascular / respiratory examinations were performed post-dose on the first day of dosing.

[0088] At the time of sacrifice, animals underwent a complete gross necropsy, including collection, weighing, fixation, and histopathological examination of the entire list of organs. Histological evaluation of the brain consisted of multiple sections encompassing the injection site (i.e., the injected lateral ventricle) and the contralateral (uninjected) lateral ventricle.

[0089] The formulations were made up fresh every four days and kept refrigerated until use according to the results of stability studies.

[0090] There were no unscheduled deaths. Body weight decreased by 5-15% over the course of the treatment period in all groups, including controls, with no significant differences in body weight between groups. Anorexia or decreased appetite was observed in the majority of animals 1 day or more after the start of dosing. The decreased appetite did not appear to correlate with either the number of doses or the dose level.

[0091] Beginning 2 hours after the first dose, dogs in all treatment groups exhibited some or all of the following symptoms, one or more times throughout the study period: passivity, elevated body temperature (greater than 101°F / 38.3°C), ear twitching, body / head / chin trembling, changes in body posture and vocalization.

[0092] Cardiovascular and respiratory function was assessed in conscious, restrained animals approximately 2 hours after the first dose, and included a 12-lead electrocardiogram (ECG), blood pressure, respiratory rate, hemoglobin pulse oximetry, capillary refill time via blood pressure cuff, and manual auscultation of the thorax to assess heart rate and lung sounds.

[0093] All animals were included in the analysis. There were no treatment-related effects on cardiovascular or respiratory parameters at any dose level.

[0094] Extensive and detailed histological evaluation was performed on multiple levels of the brains from all animals. Histopathological findings were as follows: inflammatory changes in the ventricles, adjacent neuropil, and meninges were present in the brains of all animals at the level of the injection site, and usually to a lesser extent in other regions of the cortex, cerebellum, and brainstem. These findings were also present in the brains of dogs administered aCSF and were similar in severity and extent to those seen in animals treated with vehicle (0.5% Kolliphor EL) and animals receiving any of the three dose levels of the test article. Focal hemorrhages of varying size were generally observed in most dogs in all groups receiving the drug product of Example 1 (Investigational Medicinal Product - IMP). Although absent in vehicle-treated animals (but present in one of three aCSF-treated animals), the lack of correlation with localization and dose level suggests that these hemorrhages were due to local mechanical trauma from the catheter tip rather than the result of test article toxicity. There did not appear to be any differences in the incidence or severity of inflammatory responses and hemorrhage related to the number of injections each dog received. Apart from the findings in the brain, there were no histopathological findings in any other organs.

[0095] Although some adverse effects and histopathological lesions were observed in this study, they were believed to be related to the surgical and injection procedures rather than direct vehicle or test article toxicity.

[0096] The concentrations of U0126 in the dosage formulations used in this toxicity study were higher than in the clinical formulation, except for the low dose group in dogs (see Table 4 below). Assuming instantaneous distribution of the drug to the ventricles and CSF, the C of U0126 achieved in these compartments at the lowest dose in dogs was 0.01. max is the human C at a clinical dose of 28.5 μg. maxIn medium- and high-dose animals, the estimated C max were 3.6-fold and 7.3-fold higher (Table 3).

[0097] [Table 4]

[0098] As discussed above, differences in local tolerability / toxicity of the U0126 drug product could not be discerned in the pivotal toxicity studies. max Levels (in the ventricles and CSF) were up to 7.3-fold higher than those estimated in human patients at clinical dose levels.

[0099] Therefore, the concentration and dose of the drug product of Example 1 (0.33 or 1.0 mL of a 75 μM solution) was considered safe for use in human patients.

Claims

1. Compound (I) 【Chemistry 1】 or a pharmaceutically acceptable salt thereof at a concentration of 50 to 200 μM, A pharmaceutical composition wherein the solution further comprises 0.2 to 1.0% (v / v) polyoxyethylene castor oil.

2. 2. The pharmaceutical composition according to claim 1, wherein the concentration of compound (I) in the solution is 50 to 90 μM, for example, 70 to 80 μM.

3. 3. The pharmaceutical composition of claim 2, wherein the concentration of compound (I) in the solution is 75 μM.

4. 4. The pharmaceutical composition of any one of claims 1 to 3, wherein the polyoxyethylene castor oil is Polyoxyl 35 castor oil, such as Polyoxyl 35 castor oil available under the trade name Kolliphor® EL.

5. 5. The pharmaceutical composition of claim 1, wherein the solution comprises 0.5% (v / v) polyoxyl 35 castor oil.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the solution further comprises a buffer solution.

7. The pharmaceutical composition of any one of claims 1 to 5, wherein the solution further comprises artificial cerebrospinal fluid (aCSF).

8. The pharmaceutical composition of any one of claims 1 to 7, wherein the solution further comprises dimethyl sulfoxide (DMSO).

9. The pharmaceutical composition of any one of claims 1 to 8, wherein the solution comprises one or more additional therapeutic agents.

10. Compound (I) 【Chemistry 2】 or a pharmaceutically acceptable salt thereof at a concentration of 50 to 200 μM, said solution further comprising 0.2 to 1.0% (v / v) polyoxyethylene castor oil.

11. 11. The solution of claim 10 for use as a medicine.

12. A pharmaceutical composition according to any one of claims 1 to 9 or a solution according to claim 10 for use in the treatment of ischemic damage in a human patient.

13. 13. A pharmaceutical composition or solution according to claim 12 for use in treating ischemic injury in a human patient by direct injection into the brain.

14. A method for the treatment of ischemic damage, comprising administering to a human patient a pharmaceutical composition according to any one of claims 1 to 9 or a solution according to claim 10.

15. 11. The solution of claim 10 for the manufacture of a medicament for the treatment of ischemic damage.

16. 11. A unit dose comprising the pharmaceutical composition of any one of claims 1 to 9 or the solution of claim 10, the unit dose having a volume of 1 to 10 mL, for example 7 mL.

17. A pharmaceutically acceptable solution comprising polyoxyethylated castor oil and artificial cerebrospinal fluid.

18. Compound (I) for use in the treatment of ischemic disorders 【Transformation 3】 or a pharmaceutically acceptable solvate of a salt thereof, wherein said treatment is by direct injection into the brain of a solution of said compound or a pharmaceutically acceptable salt thereof.

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