Methods for treating neurological disorders

Abstract

The present invention provides a method for treating a neurological condition comprising administering to a patient in need thereof a dose of VX-745, or a pharmaceutically acceptable composition thereof, to provide a blood concentration of between about 15 and 45 ng / mL, or between about 20 and about 40 ng / mL, or between about 25 and about 35 ng / mL, or between about 30 and about 40 ng / mL, wherein the blood concentration achieves inhibition of cytokine signaling without inhibition of cytokine production.

Description

[0001] This application is a divisional application of application number 201580037336.8, filed on July 8, 2015, having the title "Methods for treating neurological conditions". TECHNICAL FIELD

[0002] The present application relates to methods for treating neurological conditions. BACKGROUND

[0003] The intracellular enzyme p38 MAPKα has been well characterized as a regulator of proinflammatory cytokine (IL-1β and TNFα) production by macrophages and microglia and is considered a therapeutic target in Alzheimer's disease (Munoz, 2010); the main rationale being to reduce inflammatory mediators from microglia and their downstream effects on Aβ: excitotoxicity, synaptic dysfunction, and tau phosphorylation. IL-1β-p38 MAPKα can also directly modulate memory formation and cognitive function by affecting long-term potentiation / inhibition (MacAfoose, 2009; Barrientos, 2012). However, p38 MAPKα antagonists have not been developed for AD due to the unavailability of blood brain barrier (BBB) penetrating compounds previously. Given the known effects of P38 MAPK inhibitors on proinflammatory cytokine production, they have additionally been evaluated as anti-inflammatory agents in a wide range of non-CNS diseases (rheumatoid arthritis, inflammatory bowel disease, COPD). Thus, there remains an important unmet need to develop p38 MAPKα antagonists that penetrate the BBB as therapeutic agents for AD and other neurological conditions. SUMMARY

[0004] The present application is directed to the use of a pharmaceutically acceptable composition comprising VX-745 in the manufacture of a medicament for improving cognition in a patient in need thereof, wherein the patient does not have Alzheimer's disease.

[0005] The present application is also directed to the use of a pharmaceutically acceptable composition comprising VX-745 in the manufacture of a medicament for functional recovery after stroke in a subject. BRIEF DESCRIPTION OF DRAWINGS

[0006] Figure 1 Changes in latency results from the Morris Water Maze test are depicted comparing VX-745 at doses of 0.5 mg / kg, 1.5 mg / kg, and 4.5 mg / kg in aged rats.

[0007] Figure 2To depict the changes in distance results in the Morris water maze test, comparing doses of VX-745 at 0.5 mg / kg, 1.5 mg / kg, and 4.5 mg / kg in aged rats.

[0008] Figure 3 Describe the protein levels of VX-745.

[0009] Figure 4 The median plasma concentrations of VX-745 at doses of 0.5 mg / kg, 1.5 mg / kg, and 4.5 mg / kg are depicted.

[0010] Figure 5 To depict the functional recovery outcomes of limb placement and neurological scores of 7 and 20 points after ischemic stroke, VX-745 at doses of 0.5 mg / kg, 1.5 mg / kg and 4.5 mg / kg were compared in aged rats.

[0011] Figure 6 Depicting functional recovery results in a cylinder test following ischemic stroke, comparing doses of VX-745 at 0.5 mg / kg, 1.5 mg / kg, and 4.5 mg / kg in aged rats. Detailed Implementation

[0012] definition

[0013] Carrier: The term "carrier" refers to any chemical entity that can be incorporated into a composition containing an active agent (e.g., a p38 inhibitor) without significantly interfering with the stability and / or activity (e.g., the bioactivity of the agent). In some embodiments, the term "carrier" refers to a pharmaceutically acceptable carrier. An exemplary carrier in this document is water.

[0014] Combination. As used herein, the terms "combination" and related terms refer to the simultaneous exposure of a subject to two or more therapeutic agents of the present invention. For example, a pharmaceutical agent of the present invention (e.g., a p38 inhibitor) may be administered simultaneously or sequentially with another therapeutic agent in a single unit dosage form or together in a single unit dosage form. Therefore, the present invention particularly provides a dosing regimen involving the administration of at least a pharmaceutical agent of the present invention (e.g., a p38 inhibitor), another therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or mediator (which is typically associated with one or both of the p38 inhibitor and the other therapeutic agent).

[0015] Formulation. The term "formulation" refers to a composition for administration to a patient that includes at least one active agent (e.g., VX-745) and one or more carriers, excipients, or other pharmaceutical additives. Generally, particular carriers, excipients, and / or other pharmaceutical additives are selected according to knowledge in the art to achieve the desired stability, release, distribution, and / or activity of the active agent, and should be appropriate for the particular route of administration.

[0016] Moderate dose. The term "moderate dose" as used herein refers to a delivery of VX-745 into the bloodstream sufficient to inhibit p38 MAPK-mediated intracellular signaling events following activation of cytokines and other receptors, but less than a therapeutically effective amount of VX-745 to produce inhibition of anti-inflammatory effects achieved by reducing cytokine production. In some embodiments, the term "moderate dose" refers to a dose that achieves a blood concentration that is one-half, one-third, one-fourth, one-fifth, one-sixth, one-seventh, one-eighth of the blood concentration required to reduce inflammation and treat disorders other than the neurological disorders of the present application. For example, the average blood concentration of VX-745 in the treatment of rheumatoid arthritis is about 75 ng / mL, which is consistent with the whole blood IC50 of VX-745 for inhibition of cytokine production (anti-inflammatory activity) of 65-80 ng / mL. In some embodiments, a "moderate dose" of VX-745 provides a blood concentration of between about 15 and 45 ng / mL, or between 20 and about 40 ng / mL, or between about 25 and about 35 ng / mL, or between about 30 and about 40 ng / mL, wherein the blood concentration achieves inhibition of cytokine signaling but not inhibition of cytokine production.

[0017] Parenteral. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the compositions of this application can be aqueous or oleaginous suspension. These suspensions can be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

[0018] Patient. The term "patient" as used herein means a mammal to which a formulation or a composition comprising a formulation is administered, and in some embodiments includes a human.

[0019] pharmaceutically acceptable carrier, adjuvant, or vehicle. The term "pharmaceutically acceptable carrier, adjuvant, or vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the compositions of this application include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffer substances (such as phosphates), glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0020] therapeutic agent. As used herein, the phrase "therapeutic agent" refers to any agent that elicits a desired biological or pharmacological effect when administered to an organism.

[0021] therapeutically effective amount and effective amount. As used herein, and unless otherwise specified, the terms "therapeutically effective amount" and "effective amount" of an agent means an amount sufficient to provide a therapeutic benefit in the treatment, prevention and / or management of a disease, disorder or condition, e.g., to delay or minimize one or more symptoms associated with the disease, disorder or condition being treated. In some embodiments, a composition can be said to contain a "therapeutically effective amount" of an agent if it contains an amount effective under the circumstances of the treatment regimen to be administered in a single dose form. In some embodiments, a therapeutically effective amount is an amount that is statistically likely to delay or minimize, reduce the occurrence and / or grade of, one or more symptoms or side effects of a disease, disorder or condition when administered as part of a dosing regimen.

[0022] treat / Treating. The term "treat" as used herein refers to partially or completely alleviating, inhibiting, delaying onset of, reducing incidence of, effecting prophylaxis of, ameliorating, and / or lessening a disorder, disease, or condition or one or more symptoms or manifestations of the disorder, disease, or condition.

[0023] Unit Dose. The expression "unit dose" as used herein refers to a physically discrete unit of formulation suitable for administration to a subject to be treated (e.g., for a single dose); each unit contains a predetermined quantity of active agent(s) selected to produce the desired therapeutic effect in accordance with a treatment regimen, optionally together with a pharmaceutically acceptable carrier, which can be provided in a predetermined amount. The unit dose can be, for example, a volume of liquid containing a predetermined quantity of one or more therapeutic agents in an acceptable carrier (e.g., a pharmaceutically acceptable carrier), a predetermined quantity of one or more therapeutic agents in solid form (e.g., a tablet or capsule), a sustained release formulation or drug delivery device containing a predetermined quantity of one or more therapeutic agents, etc. It will be appreciated that a unit dose can contain more than one therapeutic agent in addition to the therapeutic agent(s). For example, an acceptable carrier (e.g., a pharmaceutically acceptable carrier), diluent, stabilizer, buffer, preservative, etc. can be included as described below. It will be appreciated, however, that the total daily usage of the formulations of the present application will be decided by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular subject or organism can depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the particular active compound employed; the specific composition employed; the age, body weight, general health, sex, and diet of the subject; the time of administration and rate of excretion of the particular active compound employed; the duration of the treatment; drugs and / or additional therapies used in combination or coincidentally with the particular compound employed; and like factors as are well known in the medical arts. In some embodiments, a unit dose of VX-745 is about 1 mg, 3 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, or 50 mg.

[0024] VX-745 - a p38 MAPK inhibitor

[0025] Many extracellular stimuli, including proinflammatory cytokines and other inflammatory mediators, elicit specific cellular responses through the activation of mitogen-activated protein kinase (MAPK) signaling pathways. MAPKs are serine-threonine kinases that target prolines to transduce environmental stimuli to the nucleus. Once activated, MAPKs activate other kinases or nuclear proteins, including latent transcription factors and substrates, through phosphorylation. The novel mammalian reactivating protein kinase (p38 / RK) MAPK is a stress-activated protein kinase that mediates responses to cellular stress and inflammatory signals.

[0026] Recent human genetics data indicate that a major driver of Alzheimer's disease is dysregulated microglia and neuroinflammation. Because in vitro data indicate that the IL-1 β-p38 MAPK system is a key regulator of microglia / inflammation, p38 MAPK was identified as an important therapeutic target (Munoz, 2010). P38 MAPK is also produced within neurons and appears to have a direct role in regulating intraneuronal signaling events related to neural function (McAfoose, 2009; Correa, 2012). However, the role of inhibiting p38 MAPK in chronic Alzheimer's disease models was unknown because BBB-penetrant p38 MAPK antagonists were not previously available (Munoz, 2010). It was therefore also previously unknown the relative importance and role of inhibiting inflammation relative to other roles of inhibiting p38 MAPK, such as intraneuronal signaling of IL-1 β or TNFα.

[0027] The role of p38 MAPK in various stages of inflammation has prompted the discovery of several compounds capable of inhibiting p38 (SB203580, RWJ 67657, L-167307, VX-745, RPR200765A, and others). See, e.g., Kumar et al., "p38 MAP Kinases: Key Signaling Molecules as Therapeutic Targets for Inflammatory Diseases," Nature Reviews, 2:717-726 (2003); Brown et al., "p38 MAP kinase inhibitors as potential therapeutics for the treatment of joint degeneration and pain associated with osteoarthritis," J. Inflammation 5:22 (2008), each of which is incorporated by reference in its entirety. These pharmacological inhibitors are cytokine-inhibitory anti-inflammatory drugs that cause in vitro and in vivo inhibition of lipopolysaccharide-induced tumor necrosis factor-α (TNF-α) production, and have been developed based on the putative primary pharmacological role of reducing inflammation; for example, in clinical studies, each dose was administered to achieve blood concentrations that meet or exceed the whole blood IC50 (inhibitory concentration for achieving 50% maximal effect) for inhibiting cytokines (IL-1 β or TNFα).

[0028] VX-745 is a selective small molecule inhibitor of p38 MAPK previously developed by Vertex Pharmaceuticals for the treatment of rheumatoid arthritis (RA).

[0029]

[0030] Inhibition of MAPK by VX-745 blocks the downstream synthesis of the inflammatory cytokines TNF-a and IL-1 b. The whole blood IC50 of VX-745 is 150 to 180 nM, or between 65 and 80 ng / mL (Duffy, 2011). Because VX-745 exhibited significant anti-inflammatory activity in rodent models of arthritis, Vertex initiated clinical studies in humans with rheumatoid arthritis (RA). In a Phase 2 clinical treated with 250 mg VX-745 (b.i.d) that achieved a mean plasma drug concentration of about 75 ng / mL, significant reductions in inflammatory markers and clinical improvement were demonstrated. However, patients experienced adverse events including gastrointestinal effects such as diarrhea and abdominal pain and elevations in liver transaminases. In addition, VX-745 is known to penetrate the blood brain barrier (BBB) in animals. In fact, VX-745 achieved a brain concentration that was 1.7 times that in plasma. Animals subjected to very high doses of VX-745 experienced adverse neurological effects, but these adverse events were not observed in humans. Although the proof of concept for inhibiting p38 MAPK as a therapeutic agent for RA was validated, VX-745 was discontinued in favor of the non-blood brain barrier penetrating compound VX-702, which would allow greater inhibition of p38 MAPK mediated cytokine production outside the brain for diseases such as rheumatoid arthritis without the risk of neurological side effects.

[0031] Another study with VX-745 as a reference compound in a model of arthritis demonstrated that VX-745 at a 10 mg / kg dose was not as effective as other test compounds in inhibiting paw swelling. See Chopra et al., "Pharmacological profile of AW-814141, a novel, potent, selective and orally active inhibitor of p38 MAP kinase," International Immunopharmacology, 10:467-473 (2010), which is incorporated by reference herein in its entirety.

[0032] In a model of osteoarthritis, VX-745 showed statistically significant inhibition of knee degeneration compared to control animals when administered to rats at 50 mg / kg. VX-745 was also tested in a model of hyperalgesia and showed significant inhibition of the hyperalgesic response when administered to rats at doses of 30 mg / kg, 10 mg / kg, and 3 mg / kg. Researchers found that mice exhibited hyperalgesia at doses of 3 mg / kg, 10 mg / kg, and 30 mg / kg. However, researchers observed minimal effects at a dose of 3 mg / kg. See Brown et al., "p38 MAP kinase inhibitors as potential therapeutics for the treatment of joint degeneration and pain associated with osteoarthritis," J. Inflamm., 5:22 (2008), which is incorporated by reference herein in its entirety.

[0033] Without wishing to be bound by theory, it is believed that the clinical failure of p38 inhibitors to treat chronic inflammatory conditions such as rheumatoid arthritis is due to redundancy in the inflammatory (cytokine production) pathway. The redundancy results in upregulation of feedback loops when p38 MAPK-mediated cytokine production is inhibited chronically, resulting in an overall lack of efficacy.

[0034] Methods of the invention

[0035] In a 2-week exploratory study in 26-month-old Tg2576 mice, VX-745 treated mice had lower amyloid plaque load than control animals, but otherwise the drug effect could not be assessed because of the presence of minimal inflammation. This information, combined with reports that transgenic Ab mice show a compensatory phagocytic (anti-inflammatory) microglial cell phenotype, led to the decision not to pursue further studies in mice. Instead, a well-established rat model was chosen for an in-depth dose-response study to predict human doses; where increased pro-inflammatory cytokine expression is well established and thus better reflects the inflammatory environment in human aging and AD (Barrientos, 2012). Another advantage of the rat model is that a very good pharmacokinetic-pharmacodynamic correlation between rats and humans has been previously established for VX-745 (between studies in rat adjuvant arthritis model and human RA clinical trials). A second dose-response study in rats was conducted in a model of functional recovery after ischemic stroke.

[0036] Surprisingly, in both studies, the best effects on neurological function were at the intermediate dose level, which achieved blood concentrations below those needed to suppress cytokine production, and thus could not produce anti-inflammatory effects, but were sufficient to suppress p38 MAP-mediated intracellular signaling events. More surprisingly, the effects on neurological function at the higher dose, which did produce significant anti-inflammatory effects in the brain, were less than the neurological function effects at the intermediate dose level. Thus, in some embodiments, the present application encompasses the discovery that the potency of VX-745 for inhibiting intracellular signaling following cytokine receptor activation is about twice the potency for inhibiting cytokine production; thus providing a means to administer VX-745 to achieve suppression of cytokine signaling, and concomitantly improve neurological function, without suppressing cytokine production leading to a systemic anti-inflammatory state.

[0037] As described above, in some embodiments, the present application provides a method of treating a neurological disorder in a patient in need thereof, comprising administering to the patient VX-745 in an amount sufficient to inhibit cytokine signaling, while not substantially affecting cytokine production. In some embodiments, the present application provides a method of treating a neurological disorder by administering VX-745 in an amount sufficient to inhibit IL-1 β signaling, while not substantially affecting IL-1 β production.

[0038] As used herein, the term "neurological disorder" means any one or more of Alzheimer's disease, mild cognitive impairment, vascular dementia, Lewy Body Dementia and dementia; Parkinson's Disease; functional recovery following stroke; Frontotemporal dementia (FTD) and Progressive Supranuclear Palsy (PSP) and other tauopathies; cognitive impairment; chronic tinnitus; and Huntington's Disease. In certain embodiments, the neurological disorder treated by the provided methods is selected from Alzheimer's disease, mild cognitive impairment, vascular dementia, Lewy Body Dementia and dementia; Parkinson's Disease; functional recovery following stroke; Frontotemporal dementia (FTD) and Progressive Supranuclear Palsy (PSP) and other tauopathies; cognitive impairment; chronic tinnitus; and Huntington's Disease.

[0039] As used herein, the phrase "an amount sufficient to inhibit cytokine (i.e., IL-1β) signaling, while not substantially affecting cytokine (i.e., IL-1β) production" refers to a dosage of VX-745 that results in a blood concentration in a patient that is less than half of the blood concentration required to inhibit cytokine production. In some embodiments, "an amount sufficient to inhibit cytokine (i.e., IL-1β) signaling, while not substantially affecting cytokine (i.e., IL-1β) production" is a dosage of VX-745 that results in a blood concentration in a patient that is 50% less, 60% less, 70% less, 80% less, or 90% less than the blood concentration required to inhibit cytokine (i.e., IL-1β) production.

[0040] As used herein, the phrase "while not substantially affecting cytokine (i.e., IL-1β) production" means that the method provided results in a blood concentration of VX-745 that does not inhibit cytokine production to a measurable extent.

[0041] Without wishing to be bound by any particular theory, it is understood in one aspect of the application that VX-745 achieves inhibition of cytokine signaling at much lower blood concentrations than are required to inhibit cytokine production. It is understood in another aspect of the application that VX-745 achieves positive neurological effects at lower concentrations where cytokine signaling is inhibited, but cytokine production is not affected.

[0042] In some embodiments, the present application provides a method of improving cognition comprising administering VX-745 in an amount sufficient to inhibit cytokine signaling, while not substantially affecting cytokine production. In some embodiments, the present application provides a method of improving cognition by administering VX-745 in an amount sufficient to inhibit IL-1β signaling, while not substantially affecting IL-1β production.

[0043] As used herein, the term "improving cognition" refers to any measurable improvement in cognitive decline or other cognitive symptoms of any neurological condition, such as Alzheimer's disease, mild cognitive impairment, vascular dementia, Lewy body dementia, and dementia; Parkinson's disease; functional recovery after stroke; frontotemporal dementia (FTD) and progressive supranuclear palsy (PSP) and other tauopathies; cognitive impairment; chronic tinnitus; and Huntington's disease.

[0044] In some embodiments, the present application provides a method of improving neurological function by administering VX-745 in an amount sufficient to inhibit cytokine signaling, while not substantially affecting cytokine production. In some embodiments, the present application provides a method of improving neurological function by administering VX-745 in an amount sufficient to inhibit IL-1β signaling, while not substantially affecting IL-1β production.

[0045] In some embodiments, the methods provided include administering VX-745 to a patient in need thereof at a dose sufficient to achieve a blood concentration that results in inhibition of cytokine signaling, but wherein the blood concentration is insufficient to result in inhibition of cytokine production. In certain embodiments, the methods provided include administering VX-745 to a patient in need thereof at a dose sufficient to achieve a blood concentration that results in inhibition of IL-1 β signaling, but wherein the blood concentration is insufficient to result in inhibition of IL-1 β production.

[0046] In certain embodiments, the methods provided include administering VX-745, or a pharmaceutically acceptable composition thereof, to a patient in need thereof at a dose that provides a blood concentration of between about 15 and about 45 ng / mL. In some embodiments, the methods provided include administering to a patient in need thereof a dose of VX-745, or a pharmaceutically acceptable composition thereof, to thereby provide a blood concentration of between about 20 and about 40 ng / mL, or between about 25 and about 35 ng / mL, or between about 30 and about 40 ng / mL.

[0047] In certain embodiments, the present application provides a method of treating a neurological condition, comprising administering to a patient in need thereof a dose of VX-745, or a pharmaceutically acceptable composition thereof, to thereby provide a blood concentration of between about 15 and 45 ng / mL, or between about 20 and about 40 ng / mL, or between about 25 and about 35 ng / mL, or between about 30 and about 40 ng / mL.

[0048] In some embodiments, the present application provides a method of treating a neurological condition, comprising administering to a patient in need thereof a dose of VX-745, or a pharmaceutically acceptable composition thereof, to thereby provide a blood concentration of between about 15 and 45 ng / mL, or between about 20 and about 40 ng / mL, or between about 25 and about 35 ng / mL, or between about 30 and about 35 ng / mL, wherein the blood concentration achieves inhibition of cytokine signaling without inhibition of cytokine production.

[0049] Combination therapy

[0050] In certain embodiments, the present application provides a method of treating a neurological condition, comprising administering to a subject a low dose of VX-745 and one or more additional therapeutic agents. In some embodiments, the present application provides a method of treating a neurological condition, comprising administering to a subject a therapeutically effective amount of VX-745 sufficient to inhibit cytokine signaling without substantially affecting cytokine production in combination with one or more additional therapeutic agents selected from the group consisting of cholinesterase inhibitors, N-methyl-D-aspartate antagonists, vitamin E, anti-depressants, anti-anxiety agents, anti-psychotic agents, mood stabilizers, and sleep aids.

[0051] Representative cholinesterase inhibitors include, without limitation, donepezil rivastigmine galantamine and tacrine

[0052] Representative antidepressants include, without limitation, bupropion citalopram fluoxetine mirtazapine paroxetine sertraline trazodone venlafaxine nortriptyline and desipramine

[0053] Representative anxiolytics include, without limitation, lorazepam and oxazepam

[0054] Representative antipsychotics include, without limitation, aripiprazole clozapine haloperidol olanzapine quetiapine risperidone and ziprasidone

[0055] Representative mood stabilizers include, without limitation, carbamazepine and divalproex

[0056] Representative sleep aids include, without limitation, zolpidem, zaleplon, and chloral hydrate.

[0057] Representative N-methyl-D-aspartate antagonists include, without limitation, memantine

[0058] In some embodiments, the present application provides a method of treating a neurological disorder comprising administering to a subject a therapeutically effective amount of VX-745 sufficient to inhibit cytokine signaling, while not substantially affecting cytokine production, in combination with one or more additional therapeutic agents selected from the group consisting of: exenatide varenicline, PF-04360365, rivastigmine, LY450139, ST101, bryostatin, EVP-6124, atomoxetine, HF0220, resveratrol, galantamine, PF-01913539, semagacestat, 3APS, immunoglobulin, dimebon, alpha-tocopherol, BAY 85-8101, estrogen, progesterone, ACC-001, ginko biloba, nicergoline, piracetam, NIC5-15, xaliproden (SR57746A), indomethacin, DMXB-A, LY2062430, 11-C PIB, bapineuzumab, etanercept, ramipril, interferon beta-1a, simvastatin, lipoic acid, fish oil, curcumin, PF-04447943, folate, vitamin B6, vitamin B12, leuprolide, INM-176, AH110690, tryptophan, SK-PC-B70M, BMS-708163, escitalopram, TRx0014, BAY94-9172, cerebrolysin, epigallocatechin-galate, SB-742457, lithium, rosiglitazone, divalproex, SAR110894D, PRX-03140, CX516 (Ampalex), nicotinamide, rasagiline, AC-1202 enduramide, neramexane, razadyne, NS 2330 tamibarotene, acitretin, methylphenidate, mifepristone, ZT-1, AFFITOPE AD01, AFFITOPE AD02, GSK239512, GSK933776, SR57667B, PPI-1019, MPC-7869, AZD3480, PAZ-417, solanezumab, masitinib (AB1010), BAY1006578, docosahexaenoic acid, QS-21, MNI-558, reminyl retard, flutemetamol, estradiol, medroxyprogesterone, valproate, T-817MA, AZD1446, AAB-003 (PF-05236812), modafinil, raloxifene, atorvastatin, doxycycline, trazadone, sodium oxybate, huperzine A, lutein, zeaxanthin, AC-3933, dextroamphetamine, EPAX 1050TG, SRA-333, MNI-168, CAD106, SGS742, NP031112, SSR180711C, GSI-953, prazosin, MEM 1003, AndroGel, AVE1625, cyclophosphamate, TC-5619-238, MK0249, lecozotan, circadin, MEM 3454, PPI-1019, UB 311, PF-04494700, ABT-089, LY451395, E2020, Rofecoxib, PF-03654746, EHT 0202 etazolate, DCB-AD1, ONO-2506PO, Gantenerumab, Florbetapir, ELND005, Prednisone, Novasoy, Panax ginseng, Pioglitazone, Xanomeline, ABT-288, ABT-384, Neifracetam, AQW051, Pitavastatin, Naproxen sodium Lornoxicam, AN-1792, SR57667B, Melatonin, SAM-531, MK0952, MK0677, IFN-alpha 2A, BAY 94-9172, PYM50028, Reprotinostat SR, Thalidomide, Tramiprosate, FK962, IVIG, RO5313534, Bifeprunox, LNK-754, ELND005, NSA-789, Ramelteon, Florbetaben, SRA-444, VP4896, Celecoxib, Hydrocodone, GSI-136, Zolpidem, MK3328, Metformin, CTS21166, Elontril, Ibuprofen, Posiphen tartrate, JNJ-39393406, Testosterone, BRL-049653, BMS-708163, SAM-315, Ketoconazole, Fluconazole, Warfarin, E2609, AZD0328, LY2886721, CHF 5074, E2212, Acetaminophen, LY2811376, ABT-126, Melatonin, GSK1034702, Armodafinil, Depakote, Gemfibrozil, AL-108, Levetiracetam, and Quinacrine.

[0059] Pharmaceutical composition

[0060] In some embodiments, the methods provided include administering to the patient a pharmaceutical composition comprising VX-745, and one or more therapeutic agents, and a pharmaceutically acceptable carrier, adjuvant, or vehicle. In some embodiments, the present application provides a pharmaceutical composition comprising a dose of VX-745, and one or more therapeutic agents, and a pharmaceutically acceptable carrier, adjuvant, or vehicle, wherein the dose of VX-745 results in a blood concentration of between about 5 and 45 ng / mL, between about 10 and 45 ng / mL, between about 15 and 45 ng / mL, or between about 20 and about 40 ng / mL, or between about 25 and about 35 ng / mL, or between about 30 and about 40 ng / mL. In some embodiments, the dose of VX-745 results in a blood concentration of between about 5 and 35 ng / mL, between about 10 and 30 ng / mL, between about 10 and 25 ng / mL, between about 5 and 20 ng / mL, or between about 10 and 20 ng / mL.

[0061] In certain embodiments, the pharmaceutically acceptable compositions of the present application are formulated for oral administration. The pharmaceutically acceptable compositions of the present application can be orally administered in any orally acceptable dosage form including, but not limited to, capsules, caplets, tablets, aqueous suspensions or solutions. In the case of tablets, commonly used carriers include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are required for oral use, active ingredients are combined with emulsifying and suspending agents. If desired, certain sweetening, flavoring or coloring agents can also be added.

[0062] The amount of a compound of the application that can be combined with the carrier materials to produce a composition in a single dosage form will vary depending upon the patient and the particular mode of administration. Preferably, the compositions provided should be formulated to allow the administration of a dose of VX-745 between 1-50 mg / day to a patient receiving such compositions. Examples of compositions include those formulated to allow the administration of a dose of VX-745 between 1-10 mg, 10-25 mg, or 25-50 mg per day to a patient receiving such compositions. In other embodiments of the application, the compositions include those formulated to allow the administration of a dose of the inhibitor between 3-5 mg, 5-10 mg, 10-20 mg, 20-30 mg, 30-40 mg, or 40-50 mg per day to a patient receiving such compositions. In some embodiments, the compositions are formulated to contain a dose of 1 mg, 3 mg, 5 mg, 10 mg, 20 mg, 25 mg, 30 mg, or 50 mg of active ingredient. Dosage regimens for these formulations can include, but are not limited to, single administration, once, twice or thrice daily administration, weekly administration, and monthly administration. In some embodiments, the compositions provided are formulated to provide 40 mg / day of VX-745.

[0063] It will also be appreciated that the specific dose and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the particular compound employed, the age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician and the severity of the particular disease being treated. The amount of a compound of the application in composition will also depend upon the particular compound in the composition.

[0064] Illustration

[0065] Example 1 - Morris Water Maze

[0066] Background

[0067] Recent human genetics data indicate that a major driver of Alzheimer's disease is dysregulated microglia and neuroinflammation. In addition, in vitro data indicate that the IL-1 β - p38 MAPK system can affect memory by affecting long-term potentiation (LTP) and long-term depression (LTD) in neurons (McAfoose, 2009); and p38 MAPK has been identified as a mediator of oligomeric Abeta inhibition of LTP (Li, 2011). However, the effect of inhibiting p38 MAPK in a chronic Alzheimer's disease model was unknown due to the unavailability of a blood brain barrier (BBB) penetrating p38 MAPK antagonist (Munoz, 2010). In addition, unknown was the relative importance of p38 MAPK in the following relative to cytokine signaling mediated by p38 MAPK within neurons and the effect of inhibiting p38 MAPK on the following: p38 MAPK mediated cytokine production (IL-1 β and TNFα) by microglia.

[0068] VX745 (a BBB penetrating selective p38 MAPK alpha antagonist) inhibits both IL-1 β production and signaling with two-fold higher potency for signaling. VX-745 was tested in a second supportive study conducted in a senescent rat model; and in a functional recovery model following ischemic stroke in rats following prior non-CNS clinical experience for repositioning in Alzheimer's disease.

[0069] Objectives

[0070] The primary objective of this study was to investigate whether pre-treatment with compound VX-745 mitigates cognitive deficits in senescent rats. At the end of the study, VX-745 plasma concentrations were determined in samples collected 1 hour post-dose using LC-MS / MS. In addition, PSD95 protein levels in the cortex and hippocampus and IL-1 β levels in the hippocampus were analyzed by specific ELISA assays to assess the potential anti-inflammatory effects of VX-745 in the brain.

[0071] Methods

[0072] A total of 60 female senescent (20-24 months) and 15 young adult female Fischer rats (2-3 months) were used for the experiment. Rats were treated with vehicle or 0.5, 1.5 or 4.5 mg / kg VX-745 by oral gavage twice a day until sampling (23 days). One week after starting treatment, learning and memory deficits were analyzed with the Morris Water Maze (MWM).

[0073] Test animals

[0074] All animal experiments were performed in accordance with the National Institute of Health (NIH) guidelines for the care and use of laboratory animals and were approved by the Animal Experiment Board, Finland. A total of 60 female aged (20-24 months) and 15 young (2-3 months) adult female Fischer rats (Charles River, France) were used in the experiments. The animals were housed in standard temperature (22 ± 1 °C) and in a controlled light environment (light on from 7 am to 8 pm) with ad libitum access to food and water. The animals were grouped as follows:

[0075] • Group 1 : 15 aged rats treated orally (BID) with vehicle

[0076] • Group 2: 15 aged rats treated orally (BID) with VX-745 (0.5 mg / kg)

[0077] • Group 3: 15 aged rats treated orally (BID) with VX-745 (1.5 mg / kg)

[0078] • Group 4: 15 aged rats treated orally (BID) with VX-745 (4.5 mg / kg)

[0079] • Group 5: 15 young adult control rats treated orally (BID) with vehicle

[0080] Drug delivery and rat identification

[0081] Rats were equally assigned to treatment groups based on their body weight and visible platform pre-training performance to ensure that both good and poor learners were equally present in all treatment groups. Treatments were given 1 day twice by oral gavage until sampling. The vehicle was 1% Pluronic F108 and the dosing volume was 5 ml / kg. Administration of vehicle and test compounds was performed at 7-9 am and 3-5 pm during the pre-test period. On the day of behavioral testing, treatments were given at least 1 hour before the first test trial. Test compounds were prepared and stored according to the instructions provided by the sponsor. Rats were marked accordingly with permanent marks on the tail. Notes were taken regarding the treatment group and daily treatment time.

[0082] Morris water maze

[0083] The water maze task was originally designed by Morris et al. (J Neurosci Methods. 1984; 11 : 194 47-60). Testing was performed in a large, dark tank (200 cm in diameter) filled with clear water at a temperature of 25.0 ± 1.0 °C. An escape platform (square platform: 10 x 10 cm; 1.5 cm below water surface) was placed in the middle of the NW quadrant. Arbitrary points on the rim of the pool were designated as starting positions with N, NE, E, SE, S, SW, W, NW. Rats were lowered into the pool with their nose pointing at the pool wall at a starting point. The release point (NW) adjacent to the platform location was not used.

[0084] Prior to the start of compound treatment, a visible platform pre-training was performed to determine if there were any non-cognitive performance impairments (e.g., visual impairments and / or swimming difficulties) that could impact performance on the probe or search trials. After completion of the visible platform pre-training, data were analyzed and rats were assigned to different treatment groups based on their pre-training performance. This procedure was performed to ensure that each treatment group was equally composed of both good and poor performers in the spatial version of the water maze task.

[0085] Acquisition training - Weeks 1 (days 8-11) and 2 (days 15-18): After completion of the spatial probe trials, acquisition (search) trials were performed to determine the ability of the rats to learn the spatial relationship between the distal cue and the escape platform, which remained in the same location for all search trials. The starting points were randomized (NW was not used). Rats received 4 trials per day for 4 days / week (15 minutes apart, with a maximum of 60 seconds per trial). Latency and path length (distance) were assessed.

[0086] Probe trial (day 19): A single probe trial was performed 24 hours after the last search trial to assess memory retention. The platform was removed from the pool and the rats were placed into the pool in the quadrant opposite to the one in which the platform was previously placed. Rats were allowed to swim for 60 seconds without the platform. During the probe trial, the time spent in the target quadrant and target platform annulus (a 36 cm diameter ring-shaped area around the platform) and the number of crossings of the target platform location were measured. VX-745 treatment was not positive or negative on any of the probe parameters and, therefore, is not discussed further in this application.

[0087] Endpoints, blood samples, and tissue handling for PK 257

[0088] One hour after the last vehicle or VX-745 morning dose, animals (10 animals per dosing group) were deeply anesthetized with pentobarbital and blood samples were collected by cardiac puncture. 500 μΐ of blood was collected into EDTA microtubes, centrifuged and plasma was collected. The brain was perfused with heparinized saline and the brain was collected. The right hemisphere was post-fixed by immersion in 4% PFA in 0.1 M PB for 24 hours. After a brief wash with phosphate buffered saline, the hemisphere was cryoprotected in PB containing 30% sucrose for 2-3 days, after which it was frozen in liquid nitrogen and stored at -80°C for optional future immunohistochemical analysis. The left hemisphere was dissected to obtain the ventral and dorsal cortex, hippocampus and remaining brain parts and fresh-frozen in dry ice.

[0089] PSD95 levels in the cortex and hippocampus were analyzed using an ELISA kit (Cusabio Biotech Ltd., #CSB-EL006938RA, lot O31154387), while only the levels of Il-1 β from the hippocampus were analyzed (R&D Systems, #RLB00, lot 308544).

[0090] Plasma samples were treated using acetonitrile precipitation followed by analysis of VX745 drug concentration by LC-MS / MS.

[0091] Results

[0092] Young rats treated with vehicle showed efficient learning, with MWM latency / distance rapidly decreasing in case of sequential testing. Old rats treated with vehicle appeared cognitively impaired, exhibiting longer times and distances and little improvement over the study period than young rats. Old rats treated with VX-745 exhibited greater progressive swimming times and distance reduction, which is a sign of improved cognitive performance. For both latency and distance, the 1.5 mg / kg dose of VX-745 had the greatest effect, with this group of rats' performance being approximately equal to that of young rats treated with vehicle on the last day 17 of testing. The 1.5 mg / kg dose group showed statistically significantly better performance on day 17 for both latency and distance compared to old rats treated with vehicle (p = 0.013 and 0.019, respectively). See Figure 1Additionally, there was a significantly worse performance on the first test day with respect to distance (p=0.018), and the data was also analyzed after normalizing the data by comparing the change in latency and distance from the first test day to the last test day; again, through this analysis, the aged rats treated at 1.5 mg / kg performed significantly better than the aged rats treated with vehicle, with p=0.007 for the change in latency and p=0.012 for the change in distance. See Figure 2 .

[0093] PSD95 is a protein found in the postsynaptic structure that is thought to facilitate clustering of receptors and ion channels that ensure the postsynaptic response. PSD95 levels have been shown to decrease in Alzheimer's disease and to increase in animal models when treated with anti-inflammatory agents (Frautschy, 2001). Analysis of PSD95 protein showed a statistical trend toward higher levels of PSD95 in the VX-745 high dose (4.5 mg / kg) group compared to the aged vehicle-treated group (p=0.063 according to the Mann-Whitney rank sum test).

[0094] Analysis of IL-1 β levels in the hippocampus indicated a trend toward higher levels in the aged rats. In the VX-745 high dose (4.5 mg / kg) group, IL-1 β levels were similar to those of the young rats and lower relative to the aged vehicle-treated group (p=0.038 according to the Mann-Whitney rank sum test).

[0095] The 1.5 mg / kg dose level did not have a significant effect on PSD95 or IL-1 β, and this level additionally showed a significant improvement in performance in the Morris water maze test; indicating that the effect at this dose level on the Morris water maze test level is not mediated by inhibition of cytokine production (i.e., not by anti-inflammatory action). See Figure 3 .

[0096] VX-745 plasma drug levels at 1 hour after the last dose increased in a dose-proportional manner. See Figure 4 Based on the known plasma drug profile in rats obtained from previous preclinical studies, the 1 hour post-dose level should be close to the steady state Cmean(Cavg) at steady state (C AVG ). As a prior matter, a previously unpublished study indicated that VX-745 has an IC50 in the human system for inhibition of IL-1 β production that is about half the IC50 for inhibition of IL-1 β signaling, 1.5 mg / kg reaches the drug concentration needed to block IL-1 β signaling; only 4.5 mg / kg reaches a concentration that exceeds the whole blood IC50 for IL-1 β production.

[0097] Conclusions

[0098] The present invention relates, in part, to the discovery that inhibition of p38 MAPK in animal models of chronic Alzheimer's disease has positive cognitive effects at dose levels that are insufficient to produce anti-inflammatory effects. Consistent with that finding, PK / PD correlations indicate that the effects are mediated by inhibition of IL-1 β signaling, rather than by suppression of IL-1 β production.

[0099] Example 2

[0100] Rat transient MCAO - functional recovery

[0101] The goal of this study was to investigate whether long-term 1 day twice treatment with VX-745 starting 48 hours after occlusion provides sensorimotor recovery and neuroprotection in rats subjected to 120 minutes middle cerebral artery occlusion (tMCAO) on day 0. The 7-point and 20-point neurological score tests were performed on days 1, 3, 7, 14, 21, 28, 35 and 42 after ischemia to investigate sensorimotor deficits and general condition. The cylinder test was performed on days 7, 14, 21 and 35 after ischemia and the limb placement test on days 1, 3, 7, 14, 21, 28, 35 and 42.

[0102] Infarct volume was assessed by T2-MRI 24 hours after ischemia and rats were assigned to treatment groups based on lesion data on day 2 and treatment was started on day 2 after MRI.

[0103] Methods - Male Sprague-Dawley rats were subjected to long-term 1 day twice treatment with VX-745 starting 48 hours after 120 minutes tMCAO. Lesion size was measured with MRI at 24 hours and rats were assigned to dosing groups based on lesion size and limb placement results on day 2 (tMCAO was performed on day 0). Rats were treated with 2 different doses and subjected to sensorimotor behavioral tests throughout the experiment. The 7-point and 20-point neurological score and limb placement tests were performed to investigate sensorimotor deficits and general condition before occlusion and on days 1, 3, 7, 14, 21, 28, 35 and 42 after ischemia. The cylinder test was performed before occlusion and on days 7, 14, 21 and 35 after ischemia.

[0104] Animals. A total of 45 adult male Sprague Dawley rats, purchased from Charles River Laboratories (Sulzfeld, Germany) and weighing 250-300 g were used for the experiments. The animals were housed in standard temperature (22 ± 1 °C) and in a controlled light environment (light on from 7 am to 8 pm) with ad libitum access to food and water.

[0105] Animals were grouped as follows:

[0106] ■Group A: 15 tMCAO animals were treated with vehicle twice a day (5 ml / kg, p.o.) from day 2 to day 42.

[0107] ■Group B: 15 tMCAO animals were treated with VX-745 twice a day (1.5 mg / kg, p.o.) from day 2 to day 42.

[0108] ■Group C: 15 tMCAO animals were treated with VX-745 twice a day (4.5 mg / kg, p.o.) from day 2 to day 42.

[0109] Transient MCAO

[0110] Transient focal cerebral ischemia was induced in male Sprague-Dawley rats by MCA occlusion according to Koizumi with modifications on day 0 (Koizumi et al. Jpn. J. Stroke 8: 1-8, 1986). Rats were anesthetized with 5% isoflurane (in 70% N2O and 30% O2; flow rate 300 ml / min). The concentration of anesthetic was reduced to 1.0-2.0% during surgery. The rectal temperature was maintained at 37.0 ± 1.0°C with a thermostatically controlled blanket system. After midline skin incision, the right common carotid artery (CCA) was exposed and the external carotid artery (ECA) was ligated distal to the carotid bifurcation. A 0.22 mm diameter monofilament nylon thread with a blunted tip was inserted into the internal carotid artery (ICA) 22-23 mm up to the origin of the MCA. After monofilament insertion, the CCA was occluded with a suture for 2 hours. After 120 minutes of ischemia, MCA blood flow was restored by removing the thread and suture. The wound was closed, disinfected, and the animals were allowed to recover from anesthesia. After tMCAO, rats were monitored carefully for possible post-surgical complications. After tMCAO, rats were fed with standard laboratory chow suspended in tap water. To prevent dehydration, all rats were given intraperitoneal injections of saline (4 mL per rat) once or twice a day as needed.

[0111] Drug delivery. Oral administration (5 ml / kg) of VX-745 or vehicle was initiated 48 hours after ischemia (i.e. day 2) after MRI and continued twice a day (morning and afternoon) until day 42 endpoint.

[0112] General health status and humane endpoints. Animals were monitored twice daily by laboratory personnel. In case of a significant deterioration of the general health status of the animals, rats were terminated with an overdose of CO2 and decapitated. The definition of acceptable endpoints included: no spontaneous movement and inability to drink or eat, massive bleeding, spontaneous inflammation, missing anatomical structures, swellings or tumors larger than 20 mm, and inability to recover normality within a 30 seconds period within a 24 hours observation period.

[0113] Treatment allocation

[0114] In vivo MRI acquisition was performed on all rats 24 hours after tMCAO (i.e. day 1). Absolute T2-MRI was used to determine lesion size, tissue viability (T2 within a few milliseconds) and brain edema. Rats were allocated into treatment groups based on lesion data on day 2 and treatment was initiated on day 2.

[0115] 20-point neurological score

[0116] The 20-point neurological score test was used to assess motor and behavioral deficits after ischemia. Neurological tests were performed by an unblinded investigator before stroke (baseline) and on days 1, 3, 7, 14, 21, 28, 35 and 42 after ischemia.

[0117] The following parameters were analyzed:

[0118] • Paw placement (max 4 points)

[0119] • Righting reflex (max 1 point) Visual forepaw extension (max 2 points)

[0120] • Circling (max 4 points)

[0121] • Contralateral (max 1 point)

[0122] • Grip strength (max 2 points)

[0123] • Motility (max 3 points)

[0124] • General condition (max 3 points)

[0125] • The maximum score for a normal rat is 20 points.

[0126] 7-point neurological score

[0127] In parallel, the 7-point neurological score test was used to assess motor and behavioral deficits after ischemia (modified according to Zausinger et al., 2000). Neurological tests were performed by an unblinded investigator before stroke (baseline) and on days 1, 3, 7, 14, 21, 28, 35 and 42 after ischemia.

[0128] • Grade 6: Both forelimbs are normally extended towards the floor when lifted gently by the tail. • Grade 7: Both forelimbs are normally extended towards the floor when lifted gently by the tail.

[0129] • Grade 5: The forelimb on the contralateral side of the affected hemisphere is consistently flexed, ranging from mild wrist flexion and shoulder adduction to a severe posture with complete wrist, elbow flexion and shoulder adduction with internal rotation.

[0130] • Grade 4: The functionally abnormal rat has consistently decreasing resistance to lateral push towards the paretic side.

[0131] • Grade 3: The rat circles towards the paretic side if pulled and lifted by the tail.

[0132] • Grade 2: The rat circles towards the paretic side if pulled by the tail.

[0133] • Grade 1 : The rat spontaneously circles towards the paretic side.

[0134] • Grade 0: The rat shows no spontaneous movement.

[0135] Forelimb placing test

[0136] The forelimb placing test was used to assess the sensorimotor integration of the forelimbs and hindlimbs in response to tactile and proprioceptive stimuli (de Ryck et al., 1989; Jolkkonen et al., 2000). The forelimb placing test was performed by an unblinded investigator before tMCAo (baseline) and on days 1, 5, 7, 14, 21, 28 and 35 after ischemia.

[0137] The test had 7 forelimb placing tasks which were scored as follows:

[0138] • 2 points, rat performed normally

[0139] • 1 point, rat performed delayed (> 2 seconds) and / or incompletely

[0140] • 0 points, rat performed abnormally

[0141] • Both sides of the body were tested.

[0142] In the first task, rats were suspended 10 cm above the surface of the table. Non- lesioned rats reached out with both forelimbs toward the table. In the case of the second task, the rat was held facing the table with its forelimbs resting on the edge of the table. The forelimbs were gently pulled down away from the table and recovery and limb placement were checked. Non-lesioned rats replaced both limbs on the table. The third task was identical to the second task, except that the rat could not see the table or make whisker contact because the head was held up at a 45° angle. The rat was then placed along the edge of the table to check for lateral placement of the forelimbs (fourth task) and hindlimbs (fifth task). The limbs were pulled down as in task 2 and the recovery of the limbs was scored accordingly. In the sixth task, the rats were placed with their hind end at the edge of the table with the hindlimbs resting on the edge of the table. The hindlimbs were gently pulled down (1 at a time) and away from the table. If necessary, limb recovery to the original resting position on the edge of the table was stimulated by pushing the animal toward the edge of the table. In the seventh task, the rats were placed on the edge of the table, facing away from the surface of the table. The forelimbs of the rat were placed on the edge of the table and the rat was gently pushed from behind toward the edge. The impaired rats could not maintain their grasp and the impaired limb slipped off the edge. The highest score for a normal rat was 14.

[0143] Cylinder test.

[0144] The cylinder test (modified from Schallert and Tillerson, Innovative models of CNS disease: from molecule to therapy. Clifton, NJ, Humana, 1999) was used to quantify forelimb use asymmetry when the animals stood against the walls of their home cages. The test was performed before tMCAO and on days 7, 14, and 21 after tMCAO. The rats were monitored while they were free to move in their home cages. An unaware observer scored the contacts made by each forepaw with the cage wall while standing. A total of 15-20 contacts were recorded for each animal, and the number of impaired (left) and non-impaired forelimb contacts was calculated as a percentage of total contacts.

[0145] Results

[0146] Functional recovery was analyzed for limb placement and change from baseline (day 3) to endpoint (day 42) for the 7 and 20 scores. See Figure 5In the cylinder test, changes from day 7 to day 35 were analyzed. The 1.5 mg / kg VX-745 dose group appeared to be consistently better than the 4.5 mg / kg VX-745 group. In addition, the 1.5 mg / kg VX-745 dose group showed a trend toward increased improvement in performance changes from day 7 to day 35 in the cylinder test relative to vehicle-treated animals (p=0.088, two-sided t-test); this was a trend that was not evident in the 4.5 mg / kg dose group. See Figure 6 .

[0147] Conclusions

[0148] The results regarding functional recovery as measured by limb placement, 7-point or 20-point neurological scores in this study showed that there was substantial functional improvement in vehicle-treated animals after tMCAO, and that administration of 1.5 mg / kg VX-745 or 4.5 mg / kg did not further improve functional recovery according to these measures. In the cylinder test, with vehicle treatment, minimal functional recovery was evident, and with 1.5 mg / kg VX-745, a trend toward improvement in performance changes from day 7 to day 35 on the cylinder test relative to vehicle treatment (p=0.088) was evident. For all measures, in the change analysis, the 1.5 mg / kg VX-745 dose group appeared to be consistently better than the 4.5 mg / kg VX-745 group, further supporting the notion that any positive effect of VX-745 on neurological function occurred at a dose level that did not have a measurable anti-inflammatory effect, consistent with that dose level not achieving blood concentrations that affect cytokine production; whereas dose levels that achieved blood concentrations that suppress cytokine production, and thus have anti-inflammatory properties, had less effect on neurological function.

Claims

1. Use of a compound according to the formula for the manufacture of a medicament for promoting recovery of function in a subject who has suffered a stroke. And wherein the medicament is administered to the subject 48 hours or later after onset of stroke symptoms in the subject.

2. The use according to claim 1, wherein the compound is administered once, twice or three times daily.

3. The use according to claim 1 or 2, wherein the compound is administered weekly.

4. The use according to claim 1, wherein the medicament is suitable for oral administration.

5. The use according to claim 1, wherein the medicament is administered to the subject at a dose providing a blood concentration of between 15 ng / mL and 45 ng / mL.

6. Use of a compound according to the formula for the manufacture of a medicament for promoting recovery of neurological function following stroke, and wherein the medicament is administered 48 hours or later after the onset of stroke symptoms in the subject.

7. The use according to claim 6, wherein the compound is administered once, twice or three times daily.

8. The use according to claim 6 or 7, wherein the compound is administered weekly.

9. The use according to claim 6, wherein the medicament is suitable for oral administration.

10. The use according to claim 6, wherein the medicament is administered to the subject at a dose providing a blood concentration of between 15 ng / mL and 45 ng / mL.

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