Novel solutions

Abstract

The present invention provides a pharmaceutical composition which is an aqueous solution comprising Compound (I) (also known as U0126) or a pharmaceutically acceptable salt thereof at a concentration of 50 [mu] M to 200 [mu] M, and wherein the solution further comprises 0.2% to 1.0% (v / v) polyoxyethylated castor oil. The compositions are particularly useful for the treatment of ischemic injury, in particular subarachnoid hemorrhage, in human patients by direct injection into the brain. Uses of the compositions, unit doses containing the compositions and methods of using the compositions are also disclosed. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to solutions of pharmaceutically active compounds, particularly for use in treating ischemic injury, especially ischemic injury in the brain. Specifically, the present invention relates to aqueous solutions comprising a pharmaceutically acceptable solvate of a compound designated U0126 or a salt thereof, and the use of such aqueous solutions in treating ischemic injury. Background Art

[0002] Subarachnoid hemorrhage (SAH) is an uncommon form of stroke caused by bleeding in the space around the brain. Most commonly, it occurs when a weak area (aneurysm) in a blood vessel on the surface of the brain ruptures and leaks. Blood then pools 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), which is caused by vasospasm in the brain, i.e., constriction of blood vessels and restriction of blood flow. Blood supply to various parts of the brain may be reduced to dangerous levels, thereby disrupting normal brain function.

[0004] After confirming that SAH is caused by a brain aneurysm, surgery may be recommended to repair the affected blood vessels and prevent the aneurysm from rupturing again. This can be performed using one of two main techniques: coiling and clipping. Clipping involves opening the skull to locate the aneurysm and placing a clamp around the neck of the aneurysm. Embolization involves passing a catheter through the groin through an artery supplying the brain and depositing a platinum coil within the aneurysm, causing a blood clot to form that restricts blood flow.

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

[0006] From preliminary preclinical testing, compound U0126 has shown promise for treating SAH. U0126 has the following formula:

[0007]

[0008] It is known to be an inhibitor of MEK1 and MEK2 and has been proposed for the treatment of ischemic disorders such as subarachnoid hemorrhage. Duncia et al., Bioorg. & Med. Chem. Lett. 8 (1998) 2839-2844 describe this activity. EP1,139,512 discloses the use of U0126 administered 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.

[0009] U0126 is a difficult compound to formulate and administer, being highly insoluble in most common solvents and sensitive to both acids and bases. Treating ischemic disorders requires a formulation that is soluble in physiological fluids and therefore capable of delivering the active substance directly to the brain for use in treating ischemic disorders. Intracranial injection is a desirable mode of administration, and this means that the volume of the liquid used as a carrier must be limited. Furthermore, the stability of the liquid formulation can be a problem. U0126 is known to be soluble in DMSO, but this solvent is not suitable for liquid formulations, particularly those intended for intracranial injection.

[0010] Christensen et al., Drug Delivery, 2019, 26(1), 680–688, describe 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 per mole of castor oil.

[0011] In Christensen et al., in vitro and in vivo experiments were performed to determine the optimal dose of U0126 in vitro and to test the in vivo toxicology of this dose, and to determine the synergistic effect between Cremaphor and U0126. The toxicity of different concentrations of U0126 was studied in rats. The animals were tested using three different concentration levels of U0126: low (1×10 -6 M), medium (1×10 -5 M) and high (2×10 -5 M). 15 μL was injected in each case. Neurological observations showed that low and medium doses of U0126 were well tolerated. However, at high doses, there were signs of neurological damage. Based on this, the authors concluded that U0126 in 0.5% Cremophor up to 10 -5 The concentration of 10 μM (ie, 10 μM with 0.5% Cremophor in aqueous solution) is the maximum tolerated in vivo dose for intracerebroventricular (ICV) administration.

[0012] There remains a need for improvements in the management of ischemic injury, particularly in the brain. Summary of the Invention

[0013] The present invention provides a pharmaceutical composition, which is an aqueous solution comprising compound (I) at a concentration of 50 μM to 200 μM:

[0014]

[0015] or a pharmaceutically acceptable salt thereof,

[0016] The solution also contains 0.2% to 1.0% (v / v) polyoxyethylene castor oil.

[0017] The present invention also provides a method for treating ischemic injury, which comprises administering to a human patient an aqueous solution of compound (I) described above.

[0018] The present invention also provides an aqueous solution of compound (I) described above, for use in the manufacture of a medicament for treating ischemic injury.

[0019] The present invention also provides a unit dose comprising an aqueous solution of Compound (I) described above, wherein the unit dose has a volume of 1 mL to 10 mL. DETAILED DESCRIPTION

[0020] As described above, the present invention provides a pharmaceutical composition, which is an aqueous solution comprising compound (I) at a concentration of 15 μM to 200 μM:

[0021]

[0022] or a pharmaceutically acceptable salt thereof. The compound of formula (I) is referred to as U0126. Its chemical name is 1,4-diamino-2,3-dicyano-1,4-bis(o-aminophenylmercapto)butadiene.

[0023] The present invention also provides an aqueous solution comprising Compound (I) or a pharmaceutically acceptable salt thereof at a concentration of 15 μM to 200 μM.

[0024] 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 an inorganic acid or any other acid capable of forming a salt with U0126.

[0025] U0126 forms solvates, and these solvates can be used to prepare the compositions or solutions of the present invention. Suitable solvates include those with alcohols, such as C 1-4Solvates of alkanols, such as ethanol, exist in hemiethanolate, ethanolate, and diethanolate forms and can be used.

[0026] The inventors unexpectedly found that the solution was tolerated in vivo at concentrations between 15 μM and 200 μM and that the solution was stable. Based on earlier records, 10 μM was the maximum tolerated dose and it was expected that such high concentrations would not be tolerated in vivo.

[0027] In a preferred embodiment, the solution comprises Compound (I) at a concentration of about 25 μM to about 150 μM, for example, about 25 μM to about 100 μM. More preferably, the concentration is about 50 μM to about 90 μM, for example, about 70 μM to about 80 μM. For example, the solution of the present invention comprises Compound (I) at a concentration of about 75 μM.

[0028] Given that the molar mass of U0126 is 380.5 g / mol, the solution contains Compound (I) at a concentration of about 9.5 μg / mL to about 76.1 μg / mL. More preferably, the concentration is about 9.5 μg / mL to about 57.1 μg / mL, for example, about 9.5 μg / mL to about 38.5 μg / mL. More preferably, the concentration is about 19.0 μg / mL to about 34.2 μg / mL, for example, about 26.6 μg / mL 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.

[0029] The term "about" refers to a tolerance of ±20% of the relevant value, for example ±15% of the relevant value, such as ±10% of the relevant value or ±5% of the relevant value, for example ±2% of the relevant value.

[0030] The pharmaceutical composition of the present invention offers the highly beneficial advantage that it can be administered in a sufficiently small volume (typically 0.1 mL to 5 mL, preferably less than 2 mL) in the brain without causing harm to the brain, while also providing a sufficient amount of the drug to be effective.

[0031] When administered in a volume of 0.2 ml to 5 ml, the pharmaceutical composition of the present invention will provide a patient with a dose of 0.003 μmol to 1.0 μmol of U0126. Since the molar mass of U0126 is 380.5 g / mol, the dose of U0126 is 1.14 μg to 380.5 μg. Preferably, the volume and concentration are selected to provide a dose ranging from 5 μg to 100 μg of U0126, preferably from 5 μg to 80 μg, for example, from 5 μg to 50 μg. In a more preferred embodiment, the dose is in the range of 10 μg to 40 μg. Even more preferably, the dose is in the range of 20 μg to 30 μg. For example, when a dose of 0.33 mL of a 75 μM U0126 solution is administered to a patient, the dose is 8.55 μg. When a patient is dosed with 1.0 mL of a 75 μM U0126 solution, the dose is 28.5 μg.

[0032] As described below, the course of treatment may involve administering multiple doses of a drug to a patient.

[0033] In a preferred embodiment of the present invention, the solution also includes polyoxyethylene castor oil. The polyoxyethylene castor oil is a condensate of ethylene oxide and castor oil, and some are commercially available. Such polyoxyethylene castor oil derivatives can be prepared, for example, by reacting castor oil with ethylene oxide, for example, in a molar ratio of 1:30 to 1:40, for example, 1:35. Such products are known as Macrogolycerol ricinoleate, PEG-35 castor oil, Macrogol 35 hydrogenated castor oil, and Macrogol-35 castor oil. For example, Macrogol 35 castor oil, for example, sold under the trade name EL, formerly known as "Cremophor EL" (trademark), is available commercially.

[0034] The pharmaceutical composition or solution of the present invention may comprise polyoxyethylene castor oil at a concentration of 0.2% to 1.0% (v / v). For example, the solution may comprise polyoxyethylene castor oil at a concentration of about 0.3% to about 0.7% (v / v), such as about 0.5% (v / v). For example, it may comprise polyethylene glycol 35 castor oil at a concentration of about 0.3% to about 0.7% (v / v), such as about 0.5% (v / v).

[0035] The term "solution" is used herein to describe a homogeneous mixture of a solvent (e.g., water) and a solute (particularly U0126). The mixture is passed through a filter (e.g., a 0.20 μm filter) without any of the composition remaining in the filter. In some cases, at sufficiently high magnification, the solution can be a suspension in which the first solute is surrounded by the second solute, and the second solute is in contact with the bulk solvent.

[0036] Compound U0126, in solid form, remains intact for decades under ambient conditions. Solutions as described herein have been found to remain intact for several hours at a level acceptable for pharmaceutical use. Solutions that remain intact for the time required between preparing the solution and administering it to a patient (e.g., 2 hours, e.g., 1 hour) at a level acceptable for pharmaceutical use are considered "stable" herein. It has also been found that the solution can be frozen, and degradation of the solid state occurs only very slowly.

[0037] In certain embodiments, it may be advantageous for the pharmaceutical composition or solution to further comprise a buffer solution.

[0038] In certain embodiments, the solution further comprises artificial cerebrospinal fluid (aCSF).

[0039] "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 that of human cerebrospinal fluid as closely as possible. Such fluids are sterile aqueous solutions of electrolytes including 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. Typical aCSF contains, for example, Na + : 130mM to 170mM, K + : 2mM to 5mM, Ca 2+ : 1mM to 2.5mM, Mg 2+ : 0.5 mM to 2.5 mM. Suitably, aCSF contains Na + : 140mM to 160mM, K + : 2.5mM to 4.5mM, Ca 2+ : 1mM to 2mM, Mg 2+ : 0.5 mM to 1.5 mM. More preferably, aCSF contains Na + : 145mM to 155mM, K + : 2.5mM to 4mM, Ca 2+ : 1mM to 1.5mM, Mg 2+ : 0.5 mM to 1.3 mM. Optimally, aCSF contains Na + :about 150mM, K + : 3mM to 4mM, Ca 2+ : 1.2mM to 1.5mM, Mg 2+ : 0.7mM to 1.3mM.

[0040] The counterions included in the 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.

[0041] In certain embodiments, aCSF comprises Na + : 140mM to 160mM, K + : 2.5mM to 4.5mM, Ca 2+ : 1mM to 2mM, Mg 2+ : 0.5 mM to 1.5 mM, P: 0.3 mM to 1.5 mM, and Cl - : 130mM to 165mM. Suitably, the aCSF contains Na + : 145mM to 155mM, K + : 2.5mM to 4mM, Ca 2+ : 1mM to 1.5mM, Mg 2+ : 0.5 mM to 1.5 mM, P: 0.4 mM to 1.2 mM, and Cl - : 130mM to 155mM. More preferably, aCSF contains Na + : 145mM to 155mM, K + : 3mM to 4mM, Ca 2+ : 1mM to 1.5mM, Mg 2+ : 0.7 mM to 1.3 mM, P: 0.4 mM to 1.1 mM, and Cl - : 145mM to 155mM.

[0042] A suitable aCSF contains: Na + :150mM, K + : 3mM, Ca 2 +: 1.4 mM, Mg 2+ : 0.8 mM, P: 1.0 mM, and Cl - :155mM.

[0043] In other embodiments, each 500 ml of artificial cerebrospinal fluid aCSF comprises:

[0044] 3500mg to 3800mg of sodium chloride;

[0045] 900mg to 1000mg of sodium bicarbonate;

[0046] 350mg to 450mg of dextrose;

[0047] 120mg to 180mg of magnesium sulfate.7H2O;

[0048] 120mg to 180mg of potassium chloride;

[0049] 80mg to 120mg of calcium chloride.2H2O;

[0050] 45mg to 60mg of anhydrous disodium hydrogen phosphate; and

[0051] The balance is distilled water.

[0052] In some embodiments, each 500 ml of artificial cerebrospinal fluid (aCSF) comprises:

[0053] 3600mg to 3700mg of sodium chloride;

[0054] 930mg to 990mg of sodium bicarbonate;

[0055] 375mg to 425mg of dextrose;

[0056] 140mg to 160mg of magnesium sulfate.7H2O;

[0057] 140mg to 160mg of potassium chloride;

[0058] 90mg to 110mg of calcium chloride.2H2O;

[0059] 50mg to 55mg of anhydrous disodium hydrogen phosphate; and

[0060] The balance is distilled water.

[0061] In some embodiments, each 500 ml of artificial cerebrospinal fluid (aCSF) comprises:

[0062] 3650mg to 3690mg of sodium chloride;

[0063] 950mg to 970mg of sodium bicarbonate;

[0064] 390mg to 410mg of dextrose;

[0065] 145mg to 155mg of magnesium sulfate.7H2O;

[0066] 145mg to 155mg of potassium chloride;

[0067] 95mg to 105mg of calcium chloride.2H2O;

[0068] 50mg to 55mg of anhydrous disodium hydrogen phosphate; and

[0069] The balance is distilled water.

[0070] Particularly preferably, each 500 ml of artificial cerebrospinal fluid aCSF consists of the following:

[0071] 3670mg sodium chloride;

[0072] 960mg sodium bicarbonate;

[0073] 400mg dextrose;

[0074] 150mg magnesium sulfate.7H2O;

[0075] 150mg potassium chloride;

[0076] 100mg calcium chloride.2H2O;

[0077] 53mg anhydrous disodium hydrogen phosphate; and

[0078] The balance is distilled water.

[0079] In some embodiments, each ml of artificial cerebrospinal fluid (aCSF) consists of:

[0080] 6.0mg to 8.0mg of sodium chloride;

[0081] 0.2mg to 0.4mg of potassium chloride;

[0082] 0.15mg to 0.25mg of calcium chloride.2H2O;

[0083] 0.05mg to 0.15mg of magnesium chloride hexahydrate;

[0084] 1.2mg to 2.3mg of sodium bicarbonate;

[0085] 0.05mg to 0.15mg of disodium phosphate dihydrate;

[0086] About 4.5 mg of hydrochloric acid as a 1 M solution; and

[0087] The balance is distilled water.

[0088] More preferably, each ml of artificial cerebrospinal fluid aCSF consists of:

[0089] 6.5mg to 7.5mg of sodium chloride;

[0090] 0.25mg to 0.35mg of potassium chloride;

[0091] 0.15mg to 0.25mg of calcium chloride.2H2O;

[0092] 0.05mg to 0.15mg of magnesium chloride hexahydrate;

[0093] 1.5mg to 2.0mg of sodium bicarbonate;

[0094] 0.05mg to 0.15mg of disodium phosphate dihydrate;

[0095] About 4.5 mg of hydrochloric acid as a 1 M solution; and

[0096] The balance is distilled water.

[0097] For example, each ml of artificial cerebrospinal fluid (aCSF) consists of:

[0098] 7.2mg sodium chloride;

[0099] 0.3mg potassium chloride;

[0100] 0.2mg calcium chloride.2H2O;

[0101] 0.1 mg magnesium chloride hexahydrate;

[0102] 1.8 mg sodium bicarbonate;

[0103] 0.1 mg disodium phosphate dihydrate;

[0104] About 4.5 mg of hydrochloric acid as a 1 M solution; and

[0105] The balance is distilled water.

[0106] Therefore, the present invention provides a pharmaceutical composition or solution comprising per ml:

[0107] 9.5 μg to 76.1 μg of U0126;

[0108] 3.0mg to 7.0mg of polyoxyethylene castor oil

[0109] 6.0mg to 8.0mg of sodium chloride;

[0110] 0.2mg to 0.4mg of potassium chloride;

[0111] 0.15mg to 0.25mg of calcium chloride.2H2O;

[0112] 0.05mg to 0.15mg of magnesium chloride hexahydrate;

[0113] 1.2mg to 2.3mg of sodium bicarbonate;

[0114] 0.05mg to 0.15mg of disodium phosphate dihydrate;

[0115] About 4.5 mg of hydrochloric acid as a 1 M solution; and

[0116] The balance is distilled water.

[0117] Preferably, each ml of the pharmaceutical composition or solution comprises:

[0118] 12.5 μg to 50.0 μg of U0126;

[0119] 3.5mg to 6.5mg of Cremophor oleate

[0120] 6.5mg to 7.5mg of sodium chloride;

[0121] 0.25mg to 0.35mg of potassium chloride;

[0122] 0.15mg to 0.25mg of calcium chloride.2H2O;

[0123] 0.05mg to 0.15mg of magnesium chloride hexahydrate;

[0124] 1.5mg to 2.0mg of sodium bicarbonate;

[0125] 0.05mg to 0.15mg of disodium phosphate dihydrate;

[0126] About 4.5 mg of hydrochloric acid as a 1 M solution; and

[0127] The balance is distilled water.

[0128] For example, the pharmaceutical composition or solution of the present invention consists essentially of the components listed above. More preferably, each mL of the pharmaceutical composition or solution comprises:

[0129] 19.0 μg to 38.0 μg of U0126;

[0130] 4.0mg to 6.0mg of Cremophor oleoresin

[0131] 6.5mg to 7.5mg of sodium chloride;

[0132] 0.25mg to 0.35mg of potassium chloride;

[0133] 0.175mg to 0.225mg of calcium chloride.2H2O;

[0134] 0.075mg to 0.125mg magnesium chloride hexahydrate;

[0135] 1.5mg to 2.1mg of sodium bicarbonate;

[0136] 0.075 mg to 0.125 mg of disodium phosphate dihydrate;

[0137] About 4.5 mg of hydrochloric acid as a 1 M solution; and

[0138] The balance is distilled water.

[0139] For example, the pharmaceutical composition or solution of the present invention consists essentially of the components listed above.

[0140] For example, each ml of pharmaceutical composition or solution comprises:

[0141] 28.5 μg U0126;

[0142] 5.0mg polyoxyethylene castor oil

[0143] 6.0mg sodium chloride;

[0144] 0.30mg potassium chloride;

[0145] 0.20mg calcium chloride.2H2O;

[0146] 0.10 mg magnesium chloride hexahydrate;

[0147] 1.8 mg sodium bicarbonate;

[0148] 0.10 mg disodium phosphate dihydrate;

[0149] About 4.5 mg of hydrochloric acid as a 1 M solution; and

[0150] The balance is distilled water.

[0151] For example, the pharmaceutical composition or solution of the present invention consists essentially of the components listed above.

[0152] Preferably, the pharmaceutical composition or solution of the present invention is substantially free of dimethyl sulfoxide (DMSO). However, in some embodiments, the pharmaceutical composition or solution may further comprise dimethyl sulfoxide (DMSO) in an amount sufficiently low to be tolerated in humans.

[0153] The pharmaceutical composition or solution according to the present invention may comprise 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, such as dexamethasone), a calcium channel blocker (e.g., nimodipine), or an inhibitor of an inflammatory response component (e.g., TNFα or an interleukin blocker, typically an antibody).

[0154] The solution can be prepared by a method comprising mixing the ingredients together and, if necessary, filtering (e.g., microfiltration) thereafter. It may be necessary to use a powerful dissolution technique, such as powerful stirring or shaking, or applying ultrasound. Mixing can be performed at any suitable temperature and / or pressure. Suitably, the ingredients are mixed together at a temperature of 20°C to 45°C, suitably 25°C to 40°C, and most suitably 25°C to 30°C. In addition, the ingredients are preferably mixed together at atmospheric pressure (e.g., 1 atm). Since the composition of the present invention should be sterile, the method of the present invention should be carried out under controlled conditions, for example, the method may include sterile filtration under cooling.

[0155] The solution can be prepared extemporaneously or pre-prepared and presented in unit dose or multidose form. It can be presented as a kit, i.e., as two or more separate components, wherein component (a) comprising U0126 is provided together with component (b) comprising polyoxyethylene castor oil and, optionally, component (c) aCSF. Preferably, component (a) is provided in powder form (e.g., as a powder of lyophilized U0126). The powder can then be combined with components (b) and (c) to form the solution of the present invention. Therefore, the present invention also provides a kit suitable for preparing the solution of the present invention, comprising, as separate components, (a) a powder comprising U0126 and (b) polyoxyethylene castor oil. The kit may also optionally contain (c) a pharmaceutically acceptable isotonic solution (e.g., aCSF). The kit may comprise: a dry component (such as a powder) comprising U0126; and a liquid component comprising a mixture of polyoxyethylene castor oil and a pharmaceutically acceptable isotonic solution (e.g., aCSF). The kit may additionally comprise a dry component (such as a powder) comprising U0126 and dry mass ingredients constituting a pharmaceutically acceptable isotonic solution (eg, aCSF), and a liquid component comprising polyoxyethylene castor oil.

[0156] The masses of solid components and volumes of liquid components in the kit of the invention are selected so as to provide a pharmaceutical composition or solution of the invention upon reconstitution thereof.The kit is preferably provided with instructions for preparing a pharmaceutical composition or solution of the invention as defined herein.

[0157] For example, immediately prior to use, dry U0126 is reconstituted in the presence of polyoxyethylene castor oil using a pharmaceutically acceptable isotonic solution (e.g., aCSF), which is provided with the kit or provided by the pharmacist or medical professional performing the reconstitution. In embodiments where the kit comprises a powder comprising U0126 and dry mass components constituting a pharmaceutically acceptable isotonic solution (e.g., aCSF), immediately prior to use, the powder is reconstituted using (sterile) distilled water, thereby forming a pharmaceutically acceptable isotonic solution (e.g., aCSF) in situ. The polyoxyethylene castor oil component of the kit can be added together with the distilled water or separately therefrom (e.g., before or after the addition of the distilled water).

[0158] Suitable powders may be prepared by known methods, for example by lyophilisation.Of course, the compositions of the invention will be sterile, and thus also the components provided as part of the kit will be sterile.

[0159] The present invention also provides pharmaceutically acceptable solutions comprising Cremophor ® and artificial cerebrospinal fluid. Such solutions are particularly useful in the kits of the present invention. For example, such solutions comprise 3.0 mg to 7.0 mg of Cremophor ® and aCSF. For example, per mL of such solution comprises:

[0160] 3.0mg to 7.0mg of polyoxyethylene castor oil

[0161] 6.0mg to 8.0mg of sodium chloride;

[0162] 0.2mg to 0.4mg of potassium chloride;

[0163] 0.15mg to 0.25mg of calcium chloride.2H2O;

[0164] 0.05mg to 0.15mg of magnesium chloride hexahydrate;

[0165] 1.2mg to 2.3mg of sodium bicarbonate;

[0166] 0.05 mg to 0.15 mg of disodium phosphate dihydrate; and

[0167] About 4.5 mg of hydrochloric acid, as a 1M solution,

[0168] The remainder is water.

[0169] Based on the findings described herein, the solutions of the present invention are suitable for use as medicaments.The pharmaceutical compositions and solutions described herein are particularly useful for the treatment of ischemic disorders, especially subarachnoid hemorrhage.

[0170] The present invention provides an aqueous solution comprising U0126 or a pharmaceutically acceptable solvate or salt thereof at a concentration of 15 μg to 100 μg for use as a medicament. Also provided are solutions of the preferred and exemplary forms described above for use as a medicament.

[0171] Preferably, the solution is used to treat ischemic injury in a human patient.

[0172] Prior to the present invention, U0126 had never been injected intracranially. Therefore, the present invention also provides U0126, or a pharmaceutically acceptable salt or solvate thereof, for use in treating an ischemic disorder by injecting a solution of U0126, or a pharmaceutically acceptable salt or solvate thereof, directly into the brain; and a method of treating an ischemic disorder comprising injecting a solution comprising U0126, or a pharmaceutically acceptable salt or solvate thereof, directly into the brain.

[0173] The present invention also provides a method for treating ischemic injury in a human patient, comprising administering a treatment 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.

[0174] The present invention also provides use of the solution of the present invention for the manufacture of a medicament for treating ischemic disorders. Preferably, the ischemic disorder is subarachnoid hemorrhage.

[0175] The solution of the present invention can be provided in unit dosage form. In certain embodiments, the invention provides a unit comprising a solution having a volume of 0.2 mL to 10 mL, from which a treatment dose can be obtained. In a preferred embodiment, the unit has a volume of 3 mL to 9 mL, for example 5 mL to 8 mL, for example 7 mL. Typically, the dosage volume applied to a patient is within the range of 0.1 mL to 5 mL. Preferably, it is lower than 2 mL, for example, it can be 0.2 mL to 1.5 mL, preferably 0.3 mL to 1.0 mL, for example 0.33 mL or 1.0 mL.

[0176] Example

[0177] Example 1: Preparation of a solution of U0126 in aCSF containing 0.5% Cremophor EL (Tocris Pharma)

[0178] Prepare a solution of sodium chloride, potassium chloride, calcium chloride dihydrate, and magnesium chloride hexahydrate by mixing in water (water for injection grade). Adjust the pH to 2.8 using 1 M HCl and / or 0.5 M NaOH. Prepare a second solution by mixing sodium bicarbonate and disodium phosphate dihydrate in water (water for injection grade). Mix the first and second solutions together to form the aCSF solution.

[0179] 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 and combined to provide a final volume of 3 L.

[0180] Sterile filter the solution using two Millipak 1000 0.22 μm PVDF filter units. Fill the solution into vials and seal the vials. Check the product for purity and the correct final concentration. The final concentrations of the components are shown in Table 1:

[0181] Table 1: Concentrations of components of U0126 solution in aCSF containing 0.5% Cremophor EL

[0182] Components Volume / mL U0126 28.5 μg KolliphorEL 5.0mg Sodium chloride 7.2mg potassium chloride 0.3mg Calcium chloride dihydrate 0.2mg Magnesium chloride hexahydrate 0.1mg Sodium bicarbonate 1.8mg Disodium phosphate dihydrate 0.1mg Hydrochloric acid, as a 1 M solution Appropriate dose (qs) (about 4.5 mg) water to 1ml

[0183] That is, the solution contained 28.55 μg / ml of U0126 (75 μM) and 0.5% v / v of Kolliphor EL.

[0184] Example 2: Alternative preparation of the solution of the invention

[0185] 0.79 g of Cremophor EL (trademark) was weighed into an Erlenmeyer flask and 60 mL of aCSF (available from TocrisPharma) 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 a 1.0% v / v Cremophor EL solution in aCSF containing 194 μM U0126. The solution was then placed in an ultrasonic bath maintained at 25°C to 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 obtain a 0.5% v / v Cremophor EL solution in aCSF calculated to contain 78.75 μM U0126. The actual measured content of U0126 was 67 μM (HPLC, Agilent 1100).

[0186] The mixture was then filtered using a Millipore Millex-LG, PTFE, 0.20 μm filter (SLLG025SS). The filter removed all solid material, leaving a clear solution containing a measured amount of 68 μM U0126.

[0187] Example 3: Alternative preparation of the solution of the invention

[0188] 500 μL of Cremophor EL was pipetted into an Erlenmeyer flask containing 40 mL of in-house prepared aCSF. The mixture was shaken vigorously 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 shaken vigorously for 43 minutes, resulting in an unclear solution. An additional 87 minutes of shaking failed to dissolve the remaining particles. The mixture was then sonicated for 5 minutes, whereupon the particles dissolved. An additional 10 minutes of sonication was performed to ensure complete dissolution. The resulting solution was calculated to contain 1.0% Cremophor EL and 200 μM U0126.

[0189] 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 were filtered using a Millipore sterile syringe filter (Millex-LG PTFE 0.20 μm, SLLG013SL), and the pH was measured to be 8.6. Analysis of various samples revealed U0126 levels ranging from 85 μM to 99 μM.

[0190] Biological Example 1: Trials in Human Subjects with SAH

[0191] A randomized, double-blind, placebo-controlled trial was conducted to determine the safety and tolerability of the pharmaceutical product of Example 1 (Study Medicinal Product - IMP) administered to patients as one and three ICV bolus injections over 18 hours in the early phase after subarachnoid hemorrhage (SAH). The trial included five patient groups, and they were administered the pharmaceutical product of Example 1 (Study Medicinal Product - IMP) as follows:

[0192] Cohort 1: Three patients, low-dose IMP, no placebo, single dose

[0193] Cohort 2: Four patients, medium dose, randomized 3:1 to IMP:placebo, single dose Cohort 3: Four patients, full dose, randomized 3:1 to IMP:placebo, single dose Cohort 4: Four patients, full dose, randomized 3:1 to IMP:placebo, multiple dose Cohort 5: 12 patients, full dose, randomized 3:1 to IMP:placebo, multiple dose

[0194] The study medicinal product (IMP) was a solution as described above in Example 1. It contained the active pharmaceutical ingredient U0126 at a concentration of 28.5 μg / mL. For low-dose patients, 0.33 mL of the solution was administered. For medium-dose patients, 0.6 mL of the solution was administered. For high-dose patients, 1.0 mL of the solution was administered. For multi-dose patients, three doses of 1.0 mL each were administered.

[0195] The placebo was 0.9% saline.

[0196] The trial had the following inclusion and exclusion criteria:

[0197] Inclusion criteria :

[0198] Patients who met the following criteria were eligible to participate:

[0199] 1. Male or female patients aged 18 to 80 years (inclusive).

[0200] 2. Moderate or severe SAH diagnosed by CT and caused by (or suspected to be caused by) ruptured saccular aneurysm with symptoms lasting less than 8 hours.

[0201] 3. Obtain intraventricular access within 8 hours of symptom onset.

[0202] 4. WFNS score of 1 to 5, assessed at any time after the diagnosis of SAH but before the first dose of IMP.

[0203] 5. Obtain informed consent from the trial guardian before initiating any trial-related procedures.

[0204] Exclusion criteria :

[0205] Patients were excluded from participation in the trial if they met any of the following criteria:

[0206] 1. SAH is caused by other reasons (such as trauma, rupture of spindle or mycotic aneurysm).

[0207] 2. Blood in the ventricles or brain, but not in the subarachnoid space.

[0208] 3. Expected survival <48 hours.

[0209] 4. Any serious or unstable chronic or acute concomitant disease that the PI / Representative believes will affect the safety assessment of the IMP.

[0210] 5. Any known or CT evidence of previous major brain injury or pre-existing cerebrovascular disorder that the PI / Representative believes may affect the accurate diagnosis and evaluation of SAH.

[0211] 6. Participation in any other clinical trials using experimental drugs in the past 12 weeks.

[0212] 7. Known hypersensitivity to IMP or its components.

[0213] 8. Pregnant or breastfeeding female patients. Females of childbearing potential must have a negative plasma or urine pregnancy test at screening.

[0214] Investigational Medicinal Product (IMP) Preparation and Administration :

[0215] The IMP will be prepared at the trial site by an unblinded pharmacist as detailed by the pharmacy.

[0216] Administration of IMP or placebo was performed by intracerebroventricular injection according to the following schedule shown in Table 2:

[0217] Table 2: Administration schedule of IMP or placebo

[0218]

[0219] *These doses can be taken on Day 2.

[0220] After administering 0.33 mL, 0.6 mL, or 1.0 mL of IMP, 2 mL to 5 mL of isotonic NaCl injection was also administered. This was to ensure that the entire IMP dose reached the target. The ICV connection should remain closed for 15 to 45 minutes after the injection. During the closed connection period, intracranial pressure (ICP) was monitored frequently, at the discretion of the principal investigator or their representative, to avoid excessive ICP.

[0221] For each patient, the following information was recorded as part of the trial protocol:

[0222] Demographic information (age, sex, height, weight)

[0223] Medical history and SAH diagnosis

[0224] Concomitant medication

[0225] Vital signs

[0226] Electrocardiogram: Continuous monitoring with 3-lead ECG was performed throughout the NICU stay, as per clinical practice.

[0227] Magnetic Resonance Imaging Scan: An MRI scan will be performed to document cerebral infarction at visit day 84. World Federation of Neurosurgical Societies Score: Level of consciousness and neurological deficits 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.

[0228] Glasgow Coma Scale (GCS): The GCS is used to assess the patient's level of consciousness upon hospitalization. It should be administered at least once a day.

[0229] Extended Glasgow Outcome Scale (GOS-E): The GOS-E is used to assess a patient's health status using eight categories of healthy clinical functioning: death, vegetative state, lower severe disability, upper severe disability, lower moderate disability, upper moderate disability, lower good recovery, and upper good recovery.

[0230] Modified Rankin Scale (mRS): The mRS will be used to measure the degree of disability or dependence on a caregiver for daily activities.

[0231] NIHSS: NIHSS grade for focal neurologic deficits was assessed at baseline and as a measure of disease progression up to 14 days after SAH.

[0232] Laboratory Safety Assessments: Blood samples were collected for clinical chemistry and hematology analysis. Arterial blood gas parameters were collected and sent to a certified clinical chemistry laboratory at a local laboratory and analyzed by routine analytical methods. Blood samples for laboratory safety assessments were collected before each IMP administration (within 1 hour for the second and third doses), 75 minutes (±15 minutes) after each IMP administration, and then daily for the remainder of the intensive care unit (except for blood gases, unless clinically indicated) for the remainder of the NICU patient's care.

[0233] Pregnancy test

[0234] Delayed cerebral ischemia

[0235] ·Register the length of stay in the NICU, hospital ward, or other hospital

[0236] Any exploratory procedures and assessments conducted.

[0237] At the time of filing this patent application, Cohorts 1 to 3 were complete, while Cohort 4 was partially complete. No serious adverse events (SAEs) causally related to the preparation or administration procedures of the study drug were reported (including in Cohort 4, where patients received multiple doses: three doses of 1.0 mL each, as described in the above protocol). Efficacy data will be available once the remaining phases of the study are completed.

[0238] Biological Example 2: Repeated Dose Toxicity Study in Dogs

[0239] The objective of this study was to evaluate the toxicity of the drug product of Example 1 (Investigational Medicinal Product - IMP) in beagle dogs administered once daily by ICV injection for eight days (Day 0 to Day 7), with a focus on effects in the brain. In addition, cardiovascular and CNS safety pharmacology assessments were included as an integral part of this study.

[0240] The study consists of five groups, each group of up to five beagles (up to 3 males and 2 females, see Table 3). Magnetic resonance imaging is performed on a portion of the animals to facilitate the positioning of the ventricular space. After imaging and analyzing this part, an ICV cannula is surgically implanted in one lateral ventricle of each animal. After recovery from surgery, the drug product (study drug-IMP) of Example 1 (respectively) is administered to groups 1 to 5 (respectively) with a vehicle and three different concentrations, with a dosage volume of 500 μL per dose (followed by 50 μL artificial cerebrospinal fluid). The dosage levels and the number of animals that completed the study are shown in Table 3 below:

[0241] Table 3: Dose levels and number of animals completing the study

[0242]

[0243] By slow ICV injection (about 3 μ L / sec) once a day, apply, continue eight days, a total of (at most) 8 doses. Blood plasma and serum are collected on the 0th day and the 7th day after administration. During the treatment period (the 0th day to the 7th day), the clinical signs of animals are monitored in the first hour after administration, and are monitored up to four times a day. Before starting administration and after administration on the 0th day and the 7th day, body weight and food consumption are monitored and functional observation group assessment is performed. Cardiovascular / respiratory examination is performed after administration on the first day of administration.

[0244] At sacrifice, animals underwent a complete gross necropsy, including collection, weighing, fixation, and histopathological examination of all organs. Histological evaluation of the brain included multiple sections, including the injection site (ie, the injected lateral ventricle) and the contralateral (non-injected) lateral ventricle.

[0245] Based on the results of the stability study, fresh formulations were prepared every four days and kept refrigerated until use.

[0246] There were no unplanned deaths. Body weights of all groups, including controls, decreased by 5% to 15% during the treatment period, with no significant differences in body weight between groups. Loss of appetite or decreased appetite was observed in most animals one or more days after the start of dosing. This decreased appetite did not appear to be related to the number of doses or dose level.

[0247] Beginning two hours after the first dose, dogs in all dosing groups exhibited some or all of the following signs at one or more times throughout the study: passivity, elevated body temperature (values ​​exceeding 101°F / 38.3°C), ear twitching, body / head / jaw shaking, changes in body posture, and vocalizations.

[0248] Approximately 2 hours after the first dose, cardiovascular and respiratory function were assessed in conscious, restrained animals. Examinations included a 12-lead electrocardiogram (ECG), blood pressure, respiratory rate, hemoglobin pulse oximetry, capillary refill time measured by a blood pressure cuff, and manual auscultation of the chest to assess heartbeat and lung sounds.

[0249] All animals were included in the analysis. There were no treatment-related effects on cardiovascular or respiratory parameters at any dose level.

[0250] Extensive and detailed histological evaluations were performed on multiple levels of the brain from all animals. Histopathological findings were as follows: inflammatory changes were present in the ventricles, adjacent neuropil, and meninges of the brains of all animals at the injection site level, and generally in the cortex, cerebellum, and other areas of the brainstem to a lesser extent. These findings were also present in the brains of dogs administered aCSF and were similar in severity and extent to those observed 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 sizes were commonly observed in most dogs in all groups receiving the drug product of Example 1 (study drug - IMP). Although absent in vehicle-treated animals (but present in one of the three aCSF-treated animals), the localization and lack of correlation with dose level suggest that these hemorrhages were not the result of test article toxicity, but rather due to local mechanical trauma to the catheter tip. There did not appear to be any difference in the incidence or severity of inflammatory reactions and hemorrhages associated with the number of infusions received by each dog. Apart from the findings in the brain, there were no histopathological findings in any other organs.

[0251] Although some adverse reactions and histopathological changes were observed in this study, they were considered to be related to the surgical and infusion procedures rather than direct vehicle or test article toxicity.

[0252] With the exception of the low-dose group in dogs, the concentrations of U0126 in the dose formulations used in this toxicity study were higher than those in the clinical formulation (see Table 4 below). Assuming transient distribution of the drug to the ventricles and CSF, the C of U0126 achieved in these compartments at the lowest dose in dogs was max Estimated C is similar to that achieved in humans at a clinical dose of 28.5 μg max The estimated C in dogs was the same in the medium-dose and high-dose animals. max 3.6 times and 7.3 times higher (Table 3).

[0253] Table 4: Calculated concentrations of U0126 and Kolliphor EL based on ventricular and CSF volumes in dogs

[0254]

[0255] As described above, no differences in local tolerability / toxicity of the U0126 drug product were observed in the pivotal toxicity studies. The estimated C max Levels (in ventricles and CSF) were higher than those estimated in human patients at clinically administered levels. max Levels were as much as 7.3 times higher.

[0256] Therefore, the concentration and dosage of the drug product of Example 1 (0.33 mL or 1.0 mL of a 75 μM solution) is considered safe for use in human patients.

Claims

1. A pharmaceutical composition, which is an aqueous solution comprising compound (I) at a concentration of 50 μM to 200 μM or a pharmaceutically acceptable salt thereof, The solution further comprises 0.2% to 1.0% (v / v) of polyoxyethylene castor oil. 2 . The pharmaceutical composition according to claim 1 , wherein the concentration of Compound (I) in the solution is 50 μM to 90 μM, such as 70 μM to 80 μM. 3 . The pharmaceutical composition according to claim 2 , wherein the concentration of Compound (I) in the solution is 75 μM.

4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the polyoxyethylene castor oil is polyethylene glycol 35 castor oil, such as those sold under the trade name Purchased from EL.

5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the solution comprises 0.5% (v / v) polyethylene glycol 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 according to any one of claims 1 to 5, wherein the solution further comprises artificial cerebrospinal fluid (aCSF).

8. The pharmaceutical composition according to 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. An aqueous solution comprising compound (I) at a concentration of 50 μM to 200 μM or a pharmaceutically acceptable salt thereof, The solution further comprises 0.2% to 1.0% (v / v) of polyoxyethylene castor oil.

11. The solution according to claim 10 for use as a medicament.

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

13. The 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 of treating ischemic injury, comprising administering to a human patient the pharmaceutical composition of any one of claims 1 to 9 or the solution of claim 10.

15. The solution of claim 10, for use in the manufacture of a medicament for treating ischemic injury.

16. A unit dose comprising the pharmaceutical composition according to any one of claims 1 to 9 or the solution according to claim 10, wherein the unit dose has a volume of 1 mL to 10 mL, such as 7 mL.

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

18. A compound (I) or a pharmaceutically acceptable solvate of a salt thereof, for use in the treatment of ischemic disorders by directly injecting a solution of the compound or a pharmaceutically acceptable salt thereof into the brain.

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