RU2721401C2 - Methods of treating or relieving migraine - Google Patents

Abstract

FIELD: medicine; neurology.

SUBSTANCE: present group of inventions concerns treating migraine. That is ensured by administering an effective amount of a partial NMDAR receptor agonist, CLYX-13 or a pharmaceutically acceptable salt thereof.

EFFECT: it provides the possibility of treating various migraine conditions by suppressing spreading and increasing refractory period of spreading depression in cerebral tissues.

18 cl, 11 dwg, 3 ex, 1 tbl

Description

CROSS RELATIONS TO RELATED APPLICATIONS

[0001] This application claims the benefits and priority of provisional application US No. 62/015727, filed June 23, 2014, and US provisional application No. 62/109386, filed January 29, 2015, each of which is incorporated herein in full volume by reference.

BACKGROUND

et al., Curr. Opin. Neurol., 17 (3): 289-93, 2004). РД представляет собой медленно распространяющееся угнетение электрокортикографической активности, инициируемое локальным увеличением внеклеточного калия и высвобождением глутамата, что создает самораспространяющуюся волну медленной деполяризации в больших областях коры головного мозга. Мигрень с аурой испытывают приблизительно 15-30% лиц, страдающих от мигрени.[0002] Migraine is a primary episodic pain disorder in the form of a headache associated with recurrent debilitating seizures that are often poorly controlled by existing pharmacotherapy methods focused on vascular triggers. The onset of migraine attacks is often preceded by a ciliated scotoma or migraine aura, caused by the phenomenon of cortical spreading depression (RD); see Ayata, Headache, 50: 725-30, 2010; Eikerman-Haerter et al., Curr. Neurol. Neurosci. Rep., 10: 167-73, 2010; and

et al., Curr. Opin. Neurol., 17 (3): 289-93, 2004). RD is a slowly spreading suppression of electrocorticographic activity, initiated by a local increase in extracellular potassium and the release of glutamate, which creates a self-propagating wave of slow depolarization in large areas of the cerebral cortex. Migraine with aura is experienced by approximately 15-30% of persons suffering from migraine.

[0003] The mammalian central nervous system (CNS) uses a large number of neuroactive peptides to provide specialized signaling in the brain and spinal cord, including the neuroactive peptides somatostatin, cholecystokinin, VKP (vasoactive intestinal peptide), substance P, enkephalin, neuropeptide Y (NPY) , neurotensin, TBH (thyrotropin-releasing hormone), CCK (cholecystokinin), and dynorphin (see General Description in The Biochemical Basis of Neuropharmacology, Cooper, Bloom and Roth, 5th ed., Oxford University Press, New York, 1986). A careful study of the complex signaling pathways acting in the central nervous system has led to the identification of specific receptors modulated by these neuroactive peptides, which are important therapeutic targets for various disorders associated with the central nervous system.

[0004] The N-methyl-D-aspartate receptor (NMDA) (NMDAR) is one such receptor involved in neurodegenerative disorders, including stroke-related brain cell death, convulsive syndromes, and in learning and memory processes. NMDAR also plays a central role in modulating normal synaptic transmission, synaptic plasticity, and excitotoxicity in the central nervous system. NMDAR is additionally involved in long-term potentiation (LTP). LTP is a sustained amplification of neuronal connections that underlie learning and memory processes (see Bliss and Collingridge, 1993, Nature 361: 31-39).

[0005] Two general classes of glutamate receptors in the central nervous system (CNS) have been characterized. They are metabotropic glutamate receptors, which are signal proteins belonging to the G protein coupled receptor family and ionotropic glutamate receptors (Muir and Lees, Stroke, 1995, 26, 503-513). The ionotropic class is further divided into subtypes of AMPK, kainate and NMDA receptors according to their selective ligands that activate them.

[0006] Agonistic and antagonistic compounds based on an NMDA modulating small molecule have been developed for potential therapeutic applications. However, many of them are associated with very narrow therapeutic index values and undesirable side effects, including hallucinations, ataxia, irrational behavior and significant toxicity, which limit their effectiveness and / or safety.

[0007] Thus, there remains a need for improved methods of treating migraine and other related diseases with compounds providing increased efficacy and reduced unwanted side effects.

SUMMARY OF THE INVENTION

[0008] In some embodiments of the invention, the present invention relates to methods for treating migraine, comprising administering to a patient in need of a pharmaceutically effective amount of GLYX peptide. In some embodiments of the invention, the migraine may be episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, menstrual migraine, abdominal migraine, childhood recurrent syndromes and / or cluster headache. In some embodiments of the invention, a migraine is a migraine without an aura (common migraine). In some embodiments of the invention, a migraine is a migraine with an aura (classic migraine). In some embodiments of the invention, migraine is accompanied by allodynia.

[0009] For example, the disclosed compounds can functionally interact with or modulate the action of the NMDAR glycine site for the treatment of migraine (eg episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, menstrual migraine, abdominal migraine, or cluster headache).

[0010] In some embodiments, the present invention relates to methods for treating, suppressing and / or preventing cortical spreading depression (RD), comprising administering to a patient in need thereof a pharmaceutically effective amount of GLYX peptide. In some embodiments of the invention, the present invention relates to the treatment or alleviation of long-term post-migraine complications in a patient in need thereof, comprising administering to the patient a pharmaceutically effective amount of GLYX peptide.

[0011] In some embodiments of the invention, the GLYX peptide has the following structure:

or is a pharmaceutically acceptable salt thereof or a derivative thereof having partial NMDAR agonist activity.

[0012] In one aspect, the invention relates to a method for treating migraine in a patient in need thereof, comprising administering to said patient a pharmaceutically effective amount of a compound represented by the formula:

or a pharmaceutically acceptable salt thereof.

[0013] In another aspect, the invention relates to methods for treating, suppressing and / or preventing cortical spreading depression in a patient in need thereof, comprising administering to said patient a pharmaceutically effective amount of a compound represented by the formula:

or a pharmaceutically acceptable salt thereof.

[0014] In another aspect, the invention relates to methods for treating or alleviating long-term post-migraine complications in a patient in need thereof, comprising administering to said patient a pharmaceutically effective amount of a compound represented by the formula:

[0015] or a pharmaceutically acceptable salt thereof.

[0016] As described in more detail below, drugs that are effective for administration in the aura will be useful to the patient, for example, allowing the intervention to be performed at an early stage of migraine. It is contemplated that the methods described herein are useful for the treatment of migraine with aura, comprising administering a GLYX peptide in an aura.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

[0017] Figure 1 depicts a focal pervasive depression (RD) induced by a high level of [K ⁺ ] in the CA1 field of the hippocampus slices, causing a change in glow reflecting the propagating wave of mass depolarization of neurons and glia.

[0018] Figure 2 presents data for RD induced in sections of the brain; there were no significant differences between the base regions of individual consecutive episodes of RD at the initiation point (Bonferroni multiple comparison criterion, P> 0.20), which indicates that GLYX-13 does not affect the initiation of RD.

[0019] Figure 3 presents data demonstrating that GLYX-13 increases the refractory initiation period of RD. A taxiway can be successfully called up five minutes after the previous taxiway in the control slice (control), but not called in the slice processed by GLYX-13 (GLYX-13 30 ').

[0020] Figure 4A shows that the "aura" of the RD in the form of increased luminosity diverges from the initiating pipette and propagates through the slice, and can be used to calculate the conductivity of the RD (Figure 4B).

[0021] Figure 5 presents a one-way analysis of variance (ANOVA) with repeated measurements for the analysis of 6 consecutive taxiways in order to verify the stability of speed in repeated episodes of taxiway and the possible effect of GLYX-13 on the conductivity of the taxiway.

[0022] Figure 6A shows the reduction of spines in response to two episodes of RD in a control pyramidal neuron, while Figure 6B illustrates the same process in the presence of 10 μmol / L GLYX-13; Figure 6C indicates that GLYX-13 provided restoration of the spine size after taxiway.

[0023] Figure 7A shows the distribution distance, and Figure 7B shows the rate of progression of the spreading depression (in estrogen-treated and oil-free) rats, indicating that the propagation speed was higher than in rats treated with oil.

[0024] Figure 8 shows that RD in sections of estrogen-treated rats spread further than that of rat-treated rats.

[0025] Figure 9A shows that changes in luminescence associated with RD in the rat experiment occurred with a delay in the presence of GLYX-13; Figure 9B shows that RD was induced in the presence of GLYX-13.

[0026] Figure 10 shows the effect of GLYX-13 on the rate of RD distribution in estrogen-treated rats before and after GLYX-13 (F (1.8) = 3, l; p <0.05); and comparing the preliminary exposure of GLYX-13 to the two groups (F (1.8) = 4.2; p <0.05).

[0027] Figures 11A-11B show the restoration of shock-induced learning deficits using rapastinel (3 mg / kg, intravenously (IV); 1 hour after shock) in PEL tests 24 hours after shock. Figure 11A presents data on the recovery time after exposure to a shock wave (from a latent state to normal mobility). Figure 11B shows the results of a single 3-minute session of testing positive emotional learning (PEL), conducted 24 hours after exposure to the shock wave, using an intersubjective plan. N = 4-6 per group. * P <0.05 (Figure 11A) analysis of variance, or (Figure 11B) Fisher's posterior PLSD test, rapastinel + TPGM (traumatic brain injury) / vehicle + TPGM.

DETAILED DESCRIPTION OF THE INVENTION

Migraine Classifications

[0028] Migraines were first comprehensively classified in 1988. The International Headache Society recently updated its headache classification in 2004. According to this classification, migraines are primary headaches, along with tension headaches and cluster headaches, among others.

[0029] Migraines are divided into several subclasses (some of which include further subgroups):

[0030] A migraine without an aura, or “common migraine,” includes a migraine that is not accompanied by an aura.

[0031] Migraine with an aura, or "classic migraine," usually includes a migraine accompanied by an aura. Less commonly, an aura can occur without a headache or with a non-migraine headache. Two other varieties are familial hemiplegic migraine and sporadic hemiplegic migraine, in which a person experiences migraines with aura and with accompanying motor weakness. If a close relative has the same condition, then it is called "family", otherwise it is called "sporadic." Another variety is basilar migraine, in which a headache and aura are accompanied by difficulty speaking, dizziness, tinnitus, or a number of other symptoms associated with the brain stem, but not motor weakness. This type was originally thought to be caused by spasms of the basilar artery, which feeds the brain stem. The basic principles for diagnosing migraine with aura are given, for example, in Eriksen et al., European Journal of Neurology 11: 583-591, 2004, and in the International Classification of Headache Disorders, Second Edition (ICHD-II). Subtypes of migraine with aura include varieties listed in ICHD-II, such as typical aura with migraine (IHS 1.2.1), typical aura with non-migraine headache (IHS 1.2.2), typical aura without headache (IHS 1.2.3) , familial hemiplegic migraine (IHS 1.2.4), sporadic hemiplegic migraine (IHS 1.2.5), and basilar-type migraine (IHS 1.2.6).

[0032] Infantile periodic syndromes, which are usually precursors of migraine, include cyclic vomiting (irregular intense periods of vomiting), abdominal migraine (abdominal pain, usually accompanied by nausea), and benign positional dizziness in children (irregular dizziness).

[0033] Retinal migraine includes a migraine accompanied by visual disturbances or even temporary blindness in one eye.

[0034] Migraine complications describe migraines and / or auras having an unusually long duration or an unusually high frequency, or associated with a convulsive seizure or brain damage.

[0035] Probable migraine describes conditions that have some characteristics of a migraine, but for which there is insufficient data to make a reliable diagnosis of migraine (in the presence of concomitant excessive drug use).

[0036] Chronic migraine is a complication of migraines, and is a headache that meets the diagnostic criteria of migraine and is felt over a longer period of time. More specifically, for at least 15 days / month for more than 3 months.

[0037] There are four possible phases of migraine, although not all of these phases are necessarily felt: (1) a prodrome that occurs several hours or days before a headache; (2) an aura that immediately precedes a headache; (3) a pain phase, also known as a headache phase; and (4) a postdrom, which is the effects experienced after the end of a migraine attack.

[0038] Prodromal or warning symptoms are observed in about 60% of patients with migraines, with a start in the range of two hours to two days before the onset of pain or aura. Such symptoms may include a wide range of manifestations, including: mood changes, irritability, depression or euphoria, fatigue, cravings for certain foods, muscle stiffness (especially the neck), constipation or diarrhea, and sensitivity to smell or noise. They can occur in individuals suffering from migraine with aura or migraine without aura.

[0039] An aura is a transient focal neurological phenomenon that occurs before or during a headache. It occurs gradually over several minutes and usually lasts less than 60 minutes. Symptoms can be visual, sensory, or motor in nature and many people experience several symptoms. Visual effects occur most frequently; they are observed in up to 99% of cases, and in more than 50% of cases are not accompanied by sensory or motor effects. Visual disturbances often consist of a blinking scotoma (a section of partial changes in the field of view that flickers and may impair a person’s ability to read or drive). They typically occur in the central part of the field of view and then extend to the edges in zigzag lines, which are described as similar to fortifications or castle walls. The lines are usually black and white, but some people also see color lines. Some people lose part of their field of vision (known as hemianopsia), while others experience a loss of image clarity.

[0040] The second most common type is sensory auras; they are observed in 30-40% of people with auras. Often there is a tingling sensation from one side in the hand and forearm and spreads to the area of the nose-mouth from the same side. Usually, after the tingling is completed, a numbness sensation arises with loss of a sense of position. Other symptoms of the aura phase may include: speech or language disorders, dizziness, and less commonly motor problems. Motor symptoms indicate hemiplegic migraine, and a feeling of weakness often lasts more than an hour, unlike other auras.

[0041] Aura can also occur without subsequent headache. Acephalgic migraines, also known as asymptomatic migraines, are relatively rare and include an aura and other symptoms, but without a subsequent headache (i.e., without a pain phase).

[0042] During the pain phase, the headache is usually one-sided, throbbing, and has a moderate to severe intensity. It usually occurs gradually and increases with physical activity. In more than 40% of cases, however, the pain can be bilateral, and is usually associated with neck pain. Bilateral pain is especially common in people suffering from migraines without an aura. Less commonly, pain may occur first in the back or upper part of the head. Pain usually lasts from 4 to 72 hours in adults, but in young children it often lasts less than 1 hour. The frequency of seizures is variable, from several times throughout life to several times a week, with an average of about once a month.

[0043] The pain is often accompanied by nausea, vomiting, sensitivity to light, sensitivity to sounds, sensitivity to smells, fatigue and irritability. For basilar migraine, which is a migraine with neurological symptoms associated with a brain stem or with neurological symptoms on both sides of the body, the usual effects include dizziness, fainting, and confusion. Nausea occurs in almost 90% of people, and vomiting occurs in about a third. Other symptoms may include blurred vision, stuffy nose, diarrhea, frequent urination, pallor, or sweating. Swelling or tenderness of the scalp, as well as neck stiffness, may be observed.

[0044] The effects of migraine may persist for several days after the end of the main headache; this condition is called the migraine postdrom. Many report a pain in the migraine, and some report impaired thinking for several days after the end of the headache. The patient may feel tired and experience headaches, cognitive problems, gastrointestinal symptoms, mood changes, and weakness.

GLYX Peptides

[0045] GLYX-13 is a newly developed, long-acting, fast-acting antidepressant with unprecedented modulating effect on N-methyl-D-aspartate glutamate receptor activation (NMDAR). This agent, which acts on the obligate coagonistic glycine site of the NMDA receptor necessary for its activation, normalizes the activation of this critical receptor, enhancing it when it is too low, and suppressing it if it is too high. Due to this action, GLYX-13 can enhance the induction of long-term potentiation (LTP), while suppressing LTD (long-term depression), synaptic strength, and restore normal LTP in the hippocampus sections of aging animals.

[0046] As used herein, the term "GLYX peptide" refers to a peptide having partial NMDAR glycine site agonist / antagonist activity. GLYX peptides can be obtained by well-known recombinant methods or by synthesis, for example, as described in US patent No. 5763393 and 4086196, incorporated herein by reference. In some embodiments of the invention, GLYX relates to a tetrapeptide having the amino acid sequence Thr-Pro-Pro-Thr, or L-threonyl-L-prolyl-L-prolyl-L-threoninamide.

[0047] For example, GLYX-13 relates to a compound represented by the formula:

[0048] Also provided are polymorphs, homologs, hydrates, solvates, free bases and / or suitable salt forms of GLYX-13, such as, without limitation, an acetate salt. The peptide may be in a cyclized or non-cyclized form, as further described in US patent No. 5763393. In some embodiments of the invention, the GLYX-13 analogue may comprise an insertion or deletion of a fragment of one or more of the Thr or Pro groups, such as a deletion of a CH ₂ , OH, or NH ₂ fragment. In other embodiments, GLYX-13 may optionally be substituted with one or more halogens, C ₁ -C ₃ alkyl (optionally substituted with a halogen or amino group), hydroxyl and / or amino group. Partial agonists of the NMDAR glycine site are disclosed in US Patent Nos. 5763393, 6107271, and Wood et al., NeuroReport, 19, 1059-1061, 2008, the entire contents of which are incorporated herein by reference.

[0049] It should be understood that the peptides disclosed herein may contain both natural and non-natural amino acids, for example, all natural amino acids (or their derivatives), all non-natural amino acids (or their derivatives), or a mixture of natural and non-natural amino acids . For example, one, two, three or more amino acids in GLYX-13 may each independently have a D or L configuration.

[0050] GLYX-13 may primarily affect NR2B-CO containing NMDAR, and may not exhibit classic side effects of known NMDAR modulators, such as CPC-101,606 and ketamine. In some embodiments of the invention, an anti-migraine or other therapeutic effect essentially without sedation may be produced by GLYX-13 when administered to a subject in therapeutically effective amounts. In other embodiments of the invention, GLYX-13 may not have the potential for abuse (for example, it may not be addictive).

[0051] In some embodiments of the invention, GLYX-13 may increase the phosphorylation of serine-845 GluR1 AMPA. In some embodiments of the invention, glycogen synthase kinase 3β (GSK-3β) can be activated by GLYX-13. In some cases, β-catenin levels may change after administration of GLYX-13.

[0052] In some embodiments of the invention, GLYX-13 or a composition comprising GLYX-13 may provide better in vivo activity when given intravenously (iv) and / or a concentration level in the brain compared to plasma levels.

[0053] Additionally, GLYX-13 may have a broad therapeutic index compared to glycine site antagonists such as L-701,324 or other glycine site antagonists having narrow therapeutic indices, resulting in a very narrow dose range between therapeutic effects and ataxia. For example, L-701,324 has an anticonvulsant effect in doses that cause ataxia (Bristow, et al, JPET 279: 492-501, 1996). Similarly, a number of Merz compounds are anticonvulsant in doses that cause ataxia (Parsons, et al., JPET283: 1264-1275, 1997).

Ways

[0054] In one aspect, the invention relates to a method for treating migraine in a patient in need thereof, comprising the step of administering to said patient a pharmaceutically effective amount of a compound represented by the formula:

or a pharmaceutically acceptable salt thereof.

[0055] In some embodiments of the invention, the compound is administered to a patient at a dose of from about 0.01 mg / kg to about 1000 mg / kg or from about 1 mg / kg to about 500 mg / kg of the compound.

[0056] In some embodiments of the invention, the migraine is an episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, menstrual migraine, abdominal migraine, recurrent migratory migraines, or cluster headache.

[0057] In some embodiments, the migraine is episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, or cluster headache.

[0058] In some embodiments of the invention, the migraine is a migraine with aura (classic migraine).

[0059] In some embodiments of the invention, the migraine is a migraine without an aura (common migraine).

[0060] In some embodiments of the invention, migraine is accompanied by allodynia.

[0061] In some embodiments of the invention, the method comprises the step of administering from about 1 to 10 mg / kg, from about 10 mg / kg to about 250 mg / kg, from about 20 mg / kg to about 150 mg / kg , from about 30 mg / kg to about 125 mg / kg, from about 40 mg / kg to about 110 mg / kg, from about 50 mg / kg to about 100 mg / kg, from about 60 mg / kg to about 90 mg / kg, or from about 70 mg / kg to about 90 mg / kg of the compound.

[0062] In some embodiments of the invention, the method comprises the step of administering about 1 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 50 mg / kg, about 70 mg / kg, or about 100 mg / kg of the compound.

[0063] In some embodiments of the invention, the method comprises the step of administering the compound about twice a day, about every day, once every 2 days, once every 3 days, once every 4 days, once every 5 days, about once a week about once every two weeks, or about once every four weeks.

[0064] In another aspect, the invention relates to methods for treating, suppressing, or preventing cortical pervasive depression or a disease or condition caused by cortical pervasive depression in a patient in need thereof, comprising a step in which a pharmaceutically effective amount of a compound is administered to said patient, represented by the formula:

or a pharmaceutically acceptable salt thereof.

[0065] In another aspect, the invention relates to methods for treating or alleviating long-term post-migraine complications in a patient in need thereof, comprising a step in which a pharmaceutically effective amount of a compound represented by the formula is administered to said patient:

or a pharmaceutically acceptable salt thereof.

[0066] In some embodiments, the compound is administered to a patient at a dose of from about 0.01 mg / kg to about 1000 mg / kg of the compound. In some embodiments of the invention, the compound is administered to the patient at a dose of from about 1 mg / kg to about 500 mg / kg of the compound.

[0067] In some embodiments of the invention, the method includes a step in which from about 1 mg / kg to 10 mg / kg, from about 1 mg / kg to 20 mg / kg are administered; from about 10 mg / kg to about 250 mg / kg, from about 20 mg / kg to about 150 mg / kg, from about 30 mg / kg to about 125 mg / kg, from about 40 mg / kg to about 110 mg / kg, from about 50 mg / kg to about 100 mg / kg, from about 60 mg / kg to about 90 mg / kg, or from about 70 mg / kg to about 90 mg / kg of the compound.

[0068] In some embodiments of the invention, the method comprises the step of administering about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 50 mg / kg, about 70 mg / kg, 1 mg / kg 5 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg or about 100 mg / kg of the compound.

[0069] In some embodiments of the invention, the method comprises the step of administering the compound about twice a day, about every day, once every 2 days, once every 3 days, once every 4 days, once every 5 days, about once a week , or about once every two weeks, or, for example, once a month.

[0070] In some embodiments of the invention, the method further includes a step in which the compound is co-administered with an opioid, antidepressant, antiepileptic agent, non-steroidal anti-inflammatory drug (NSAID), serotonin receptor agonist 5HT1B / 1D, N-methyl-D-aspartate antagonist or anti-inflammatory compound.

[0071] Activation of NMDAR may promote or even be essential for the occurrence of RD in many experimental situations. Thus, compounds that prevent the excessive activation of NMDAR can act as important new therapies to reduce, and even prevent, the initial manifestations of migraine attacks. For example, GLYX-13, described below, inhibits the production and distribution of RD in in vitro hippocampal slices. In some experiments, GLYX-13 can completely prevent the induction of RD by locally increasing the extracellular concentration of potassium, and / or can slow down the rate of propagation of RD or even completely block it. In addition, GLYX-13 can improve the return of dendritic spines to their original sizes after RD. For example, methods for treating migraine in patients prophylactically and / or with exacerbation are provided herein. Such administration may reduce the severity (or, in some embodiments, stop) of migraine attacks in patients.

[0072] In some embodiments of the invention, the patient is a human. Alleged patients include female patients and / or adolescent patients.

[0073] Also provided herein are methods for treating migraine (for example, episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, menstrual migraine, abdominal migraine, recurrent childhood syndromes, or cluster headache). patients, or treatment of refractory migraine, for example, patients suffering from a migraine that does not respond and / or does not respond to appropriate courses of treatment with at least one or at least two other compounds or therapeutic agents. For example, this document provides methods for treating migraine (e.g., episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, menstrual migraine, abdominal migraine, childhood recurrent syndromes, or cluster headache) in an unresponsive patient comprising the following steps in which a) optionally identifying the patient as treatment refractory, and b) administering an effective dose of GLYX-13 to said patient. In some embodiments of the invention, the migraine is a migraine with an aura.

[0074] In this document, an embodiment of the invention provides methods for the acute treatment of migraine (for example, episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, menstrual migraine, abdominal migraine, childhood periodic syndromes, or cluster headaches pain) in a patient in need thereof, comprising a step in which an effective amount of the GLYX peptide is administered, for example, in a single dose. For example, this document provides, in an embodiment of the invention, methods for treating migraine in a patient in need thereof at the beginning of a migraine attack, comprising a step in which an effective amount of GLYX-13 is acutely administered (e.g., a single dose). Such methods can relieve the patient of at least one migraine symptom for a period of about 2 weeks or less, 1 week or less, 1 day or less, or 1 hour or less (for example 15 minutes or less, half an hour or less) after said administration . In some embodiments of the invention, such methods can relieve the patient of at least one migraine symptom for a period of about 1 day or more, 1 week or more, or 2 weeks or more after said administration. For example, a method is provided herein comprising a step in which an effective amount of a GLYX peptide is administered to a patient suffering from migraine, wherein said patient substantially eliminates at least one migraine symptom much earlier after the first administration of the GLYX peptide, compared to the same the patient with the introduction of another drug for the treatment of migraine. A person skilled in the art will understand that such acute administration methods may be preferred in a hospital setting or for outpatients. The methods described herein may also be useful for treating allodynia that occurs with migraine with aura.

[0075] The methods of the present invention can also be used in the treatment of patients with depression, suffering from traumatic brain damage, epilepsy, or at risk of stroke. For example, this document provides, in an embodiment of the invention, a method for treating traumatic brain damage, comprising the step of administering an effective amount of a GLYX peptide, such as GLYX-13. In another embodiment, a method of treating epilepsy is provided, comprising the step of administering an effective amount of a GLYX peptide, such as GLYX-13.

[0076] In addition to cardiovascular conditions, persons suffering from migraine with aura may have an increased risk of other neurological and / or physiological conditions and disorders. It has been shown, for example, that the coincidence of migraine with an aura with deep depression or attempted suicide increases the risk of developing an unprovoked seizure (Hesdorffer et al., Epilepsy Res. 75 (2-3): 220-223, 2007). Other conditions associated with migraine with aura include significantly increased markers of NO activity, an increased incidence of depression, and a significantly higher correlation of the genetic biomarker with stroke compared to the general population (Etminan et al., MJ 330 (7482): 63 , 2005). A person suffering from a migraine having, or having an increased risk of, any of these conditions may take medications to treat or manage these diseases, and such medications may undesirably interact with current medications for treating migraine with aura. As shown in this document, some of these conditions are contraindicated in tryptanic therapy (e.g., stroke and sumatriptan therapy). Furthermore, the U.S. Food and Drug Administration (FDA) issued a medical leaflet in 2006 regarding serotonin syndrome, a life-threatening condition that can occur when triptan is used with certain antidepressants that are inhibitors of serotonin reuptake (SSRI) or selective serotonin / norepinephrine reuptake inhibitors (SNRI). Accordingly, the methods described herein can be used to treat patients with depression or patients suffering from, or having an increased risk of, stroke.

Dosage

[0077] the Dosage of any compositions in accordance with the present description will vary depending on the symptoms, age and body weight of the patient, the nature and severity of the disorder requiring treatment or prevention, route of administration and form of the composition according to the invention. Any of the compositions of the invention may be administered in a single dose or in multiple doses. Dosages of the compositions in accordance with the present description can be easily determined by methods known to those skilled in the art, or as described herein. In general, satisfactory results can be obtained by administering the compound to humans in a daily dosage of, for example, from 0.05 mg to 3000 mg (in terms of solid form), for example, from about 10 mg to about 500 mg, or for example from about 1 to about 200 mg / kg. Dosage ranges include, for example, values in the range of 10-1000 mg (e.g., 50-800 mg). In some embodiments, 50, 100, 150, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of compound are administered. . Alternatively, the dosage amount can be calculated using the patient’s body weight. For example, the dose of a compound, or pharmaceutical composition thereof, administered to a patient may be in the range of 1-500 mg / kg (e.g., 5-250 mg / kg). In typical non-limiting embodiments of the invention, the dose may have a value in the range of 5-200 mg / kg (for example, 1, 2, 2.5, 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 mg / kg) or in the range of 15-100 mg / kg (e.g. 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180 , 190 or 200 mg / kg). In typical non-limiting embodiments of the invention, the dose may have a value in the range of 1-15 mg / kg, 50-100 mg / kg, 60-90 mg / kg, or 70-80 mg / kg.

[0078] GLYX-13 can provide a high therapeutic index. For example, GLYX-13 may be therapeutically effective when given intravenously (iv) or subcutaneously in a range of from about 1 to about 10 mg / kg, from about 10 to about 200 mg / kg, for example, about 30 mg / kg, about 75 mg / kg, or about 100 mg / kg. In some embodiments of the invention, ataxia does not occur, for example, at a dose of 500 mg / kg intravenously.

[0079] The therapeutically effective amount of GLYX peptide required for use in therapy varies depending on the form of the condition being treated, the desired length of treatment time, age and condition of the patient, and is ultimately determined by the attending physician. The desired dose may conveniently be administered as a single dose that remains effective for two weeks, one week, 6, 5, 4, 3, 2, or 1 day, or as multiple doses that are administered at suitable intervals, for example, as two, three, four or more multiple doses per day.

[0080] There may be a need to determine the effective dose or amount, and any possible effects, affecting the determination of the timing of administration of a composition for any particular composition in accordance with this document. This can be accomplished using a routine experiment, as described herein, using one or more groups of animals (preferably at least 5 animals in a group), or by conducting human trials, if necessary. The effectiveness of any composition according to the invention and the method of treatment or prophylaxis can be assessed by administering the composition and assessing the effect of administration by measuring one or more relevant indicators, and comparing the values of these indicators after treatment with the values of the same indicators before treatment.

[0081] The exact time of administration and the amount of any particular composition of the invention that provides the most effective treatment for a given patient will depend on the activity, pharmacokinetic characteristics and bioavailability of the composition of the invention, the physiological condition of the patient (including age, gender, type and stage of illness, general physical condition, susceptibility to a given dosage and type of drug treatment), route of administration, etc. The recommendations given in this document can be used to optimize treatment, for example, determining the optimal time of administration and / or amount, not beyond the scope of routine experiments, which include monitoring the condition of the subject and adjusting the dosage and / or time of administration.

[0082] During treatment of a subject, a patient's health can be monitored by measuring one or more relevant indicators at predetermined time points during the treatment period. Treatment, including composition, amounts, time of administration and technology for preparing the composition, can be optimized in accordance with the results of such a control. You can conduct periodic repeated examinations of the patient to determine the degree of improvement by measuring the same parameters. Based on such repeated determinations, the amount (s) of the composition to be administered according to the invention and possibly the time of administration can be adjusted.

[0083] Treatment may begin with low dosages less than the optimal dose of the compound. After this, the dosage may be increased in small steps until an optimal therapeutic effect is achieved.

[0084] The use of the compositions of the invention may result in a reduction in the required dosage of any individual agent included in the compositions, since the onset and duration of the effect of different agents may be imposed.

[0085] The toxicity and therapeutic efficacy of the compositions of the invention can be determined using standard pharmaceutical procedures in cell cultures or experimental animals, for example, to determine LD50 and ED50.

[0086] Data from cell culture analyzes and animal studies can be used in formulating compositions in a dosage range intended for humans. The dosage of any composition of the invention is preferably in the range of blood circulation concentrations, including the ED50, with little or no toxicity. The dosage may vary in this range depending on the dosage form used and the route of administration used. For compositions in accordance with this document, a therapeutically effective dose can be first estimated based on analyzes on cell cultures.

[0087] In some embodiments of the invention, the GLYX peptide is administered as a prophylactic agent (ie, prior to the prodrome phase). In some embodiments of the invention, the GLYX peptide is administered during the prodrome. In some embodiments of the invention, the GLYX peptide is administered in an aura. “With aura” means any time after the appearance of the aura and before the onset of migraine pain. In some embodiments of the invention, the GLYX peptide is administered after the onset of migraine pain. In some embodiments of the invention, the GLYX peptide is administered during the postdrom, for example, to reduce its symptoms.

[0088] The term "prevent," as used herein, refers to prophylactic treatment or treatment that prevents one or more of the symptoms or conditions of a disease, disorder, or conditions described herein (eg, pain or migraine with aura and with with or without allodynia). Preventive treatment can be started, for example, before ("pre-exposure prophylaxis") or after ("post-exposure prophylaxis") an event preceding the onset of a disease, disorder or condition (for example, with a migraine aura). A prophylactic treatment, comprising the step of administering the GLYX peptide described herein, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition thereof, may be acute, short-term or chronic. Dosages administered may vary during prophylactic treatment. See also: Kaniecki et al., "Treatment of Primary Headache: Preventive Treatment of Migraine." B: Standards of Care for Headache Diagnosis and Treatment. Chicago (IL): National Headache Foundation; 2004, p. 40-52.

Songs

[0089] GLYX peptides in accordance with this document can be introduced using various means, depending on their intended use, as is well known in the art. For example, if the compositions in accordance with this document must be administered orally, they can be prepared in the form of tablets, capsules, granules, powders or syrups. Alternatively, the compositions in accordance with this document can be administered parenterally in the form of injections (intravenous, intramuscular or subcutaneous), drugs for drip infusion, or suppositories. For use by administration through the mucous membrane of the eye, compositions in accordance with this document can be prepared in the form of eye drops or eye ointments. These compositions can be prepared by conventional means and, if desired, the compositions can be mixed with any conventional additive, such as an excipient, a binder, a disintegrant, a lubricant, a drug modifier, a solubilizer, a suspending agent, an emulsifier or a coating agent .

[0090] DNA encoding GLYX peptides included in the expression vector can also be introduced using any known administration method for expressing GLYX peptides in vivo.

[0091] In the compositions of this invention, wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavoring agents, may be included in the formulation of the composition. and flavors, preservatives and antioxidants.

[0092] The compositions of the invention may be suitable for oral, topical (including buccal and sublingual), rectal, vaginal, aerosol, and / or parenteral administration. The compositions may conveniently take the form of unit dosage forms and may be prepared by any means well known to those skilled in the pharmaceutical art. The amount of composition that can be combined with the material of the carrier to obtain a single dose varies depending on the subject receiving the treatment and the particular route of administration.

[0093] Methods for preparing such compositions include the step of combining the compositions as described herein with a carrier and, optionally, one or more accessory ingredients. In general, compositions are prepared by uniformly and thoroughly mixing agents with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, molding the product.

[0094] Compositions suitable for oral administration may be in the form of capsules, starch capsules, pills, tablets, lozenges (based on a flavoring substance, usually sucrose, and gum arabic or tragacanth), powders, granules, or a solution or suspension in aqueous or a non-aqueous liquid, or an oil-in-water or water-in-oil liquid emulsion, or an elixir or syrup, or lozenges (using an inert base such as gelatin and glycerin, or sucrose and gum arabic), each of which contains a predetermined amount compositions of the invention as an active ingredient. Compositions in accordance with this document can also be entered in the form of a bolus, electuary or paste.

[0095] In solid oral dosage forms (capsules, tablets, pills, dragees, powders, granules and the like), the composition of the invention is mixed with one or more pharmaceutically acceptable carriers such as sodium citrate or dicalcium phosphate, and / or any of the following materials: (1) fillers or excipients, such as starches, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders, such as, for example, carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and / or gum arabic; (3) wetting agents such as glycerin; (4) disintegrants, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; (5) dissolution retardants such as paraffin; (6) absorption accelerators such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl (acetyl) alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and (10) coloring matter. In the case of capsules, tablets and pills, the compositions may also contain buffering agents. Solid compositions of a similar type can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

[0096] A tablet may be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using a binder (e.g. gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, preservative, disintegrant (e.g. sodium starch glycolate or crosslinked sodium carboxymethyl cellulose), a surface active or dispersing agent. Molded tablets may be prepared by molding in a suitable machine a mixture of the composition of the invention moistened with an inert liquid diluent. Tablets and other solid dosage forms, such as dragees, capsules, pills and granules, may optionally be cut or prepared with coatings or shells, such as enteric coatings and other coatings well known to those skilled in the art of preparing pharmaceutical compositions.

[0097] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the compositions of the present invention, liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular cottonseed, peanut, corn, germ oil, olive, castor and sesame oils), glycerin, tetrahydrofuryl alcohol, polyethylene glycols and esters fatty acids , cyclodextrins and mixtures thereof.

[0098] Suspensions, in addition to the compositions of the invention, may contain suspending agents, such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, meta-aluminum hydroxide, bentonite, agar-agar and tragacanth, and their mixtures.

[0099] Compositions for rectal or vaginal administration can be prepared as a suppository, which can be made by mixing the composition of the present invention with one or more suitable non-irritant excipients or carriers, including, for example, cocoa butter, polyethylene glycol, suppository wax or salicylate, which are solid at room temperature but liquid at body temperature and therefore will melt in body cavities and release the active agent. Compositions suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray compositions containing carriers that are considered suitable for use in the art.

[00100] Dosage forms for transdermal administration of the compositions of the invention include powders, sprays, liquid ointments, pastes, creams, lotions, gels, solutions and patches.

[00101] Compositions for topical administration to the eye of the present invention may take the form of solutions, gels, liquid ointments, suspensions or solid inserts formulated so that a single dose contains a therapeutically effective amount of the active ingredient or some multiple thereof, in the case of combination therapy .

[00102] Pharmaceutical compositions of the present invention suitable for parenteral administration include the composition of the invention in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted to sterile solutions or dispersions for injection immediately prior to use, and which may contain antioxidants, buffers, bacteriostatic agents, dissolved substances that make the composition isotonic with the blood of the intended recipient, or suspending agents or thickeners.

[00103] Examples of suitable aqueous and non-aqueous vehicles that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerin, propylene glycol, polyethylene glycol and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate, and cyclodextrins. The desired fluidity can be maintained, for example, by using coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants.

Combination therapy

[00104] Any GLYX peptides described herein (eg, GLYX-13) can be used individually or in combination with other agents to treat or prevent any diseases or conditions described herein. For example, in some combination therapies, dosages of one or more therapeutic compounds can be reduced compared to standard dosages when administered separately.

[00105] The methods of the invention also include the step of co-administering a GLYX peptide with an opioid, antidepressant, antiepileptic agent, non-steroidal anti-inflammatory drug (NSAID), serotonin receptor agonist 5HT1B / 1D, antagonist. N-methyl-D-aspartate, or anti-inflammatory compound.

[00106] The description of the present invention relates, in some embodiments, to the use of a GLYX peptide or peptides, alone or in combination with one or more other antidepressant treatment methods, such as tricyclic antidepressants, a monoamine oxidase inhibitor (MAO-I), a selective re-inhibitor serotonin absorption (SSRI), and double and triple absorption inhibitors and / or anxiolytic drugs, for the manufacture of medicines for the treatment of migraine (e.g. episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, menstrual migraine migraines, childhood periodic syndromes, or cluster headache). Typical examples of drugs that can be used in combination with the GLYX peptide include anafranil, adaline, aventil, elavil, norpramine, pamelor, pertofran, sinequan, surmontyl, tofranil, vivactyl, parnate, nardil, marplan, celex, lexapro, luvox, Paxil, Prozac, Zoloft, Wellbutrin, Effexor, Remeron, Cymbalta, Desirel (Trazodone) and Ludiomill.

[00107] In some embodiments of the invention, the opioid is selected from the group consisting of alfentanil, butorphanol, buprenorphine, dextromoramide, desicin, dextropropoxyphene, codeine, dihydrocodeine, diphenoxylate, ethorphine, fentanyl, hydrocodone, hydromorfone, keteromede, ketomedo meptazinol, methadone, morphine, morphine-6-glucuronide, nalbuphine, naloxone, oxycodone, oxymorphone, pentazocine, pethidine, pyritramide, propoxyphene, remifentanil, sulfentanil, tilidine and tramadol.

[00108] In some embodiments of the invention, the antidepressant is selected from the group consisting of adinazolam, alaproclath, amineptin, a combination of amitriptyline / chlordiazepoxide, atipamezole, azamianserin, basinaprine, befuralin, bifemelan, binodalin, bipenamol, cyroformin, cyanocaronamine, caripaminazine, caripaminazin, caricaminophenamine, caripaminazine, caricaminophenamine, caricaminophenamine, caripaminazin , citalopram, clemeprol, clovoxamine, clazepinyl, deanol, demexiptilin, dibenzepine, dotiepine, droxidopa, ennexin, estazolam, etoperidone, femoxetine, fengabin, fesolamine, linofinoxinepinolipindepinolipinolipindepinolipindepinolipindepinolipindeinolipinolide , metapramine, metralindole, mianserin, milnacipran, minaprina, mirtazapine, montirelin, nebracetam, nefopam, nialamide, nomifenzin, norfluoxetine, orotirelin, oxaflozan, pinazepam, pirindolin, pirotinulipyrindyrindyrilin tyrindyrolimine, siripinuliprindiprolipin , tozalinone, tyroliberin, tianep tina, tiflucarbin, trazodone, tofenacin, tofisopam, toloxatone, tomoxetine, veralipride, viloxazine, vikvalin, zimelidine and zometapine.

[00109] In some embodiments, the antiepileptic agent is selected from the group consisting of carbamazepine, flupirtine, gabapentin, lamotrigine, oxcarbazepine, phenytoin, retigabin, topiramate, and valproate.

[00110] In some embodiments, a non-steroidal anti-inflammatory drug (NSAID) selected from the group consisting of acemetacin, aspirin, celecoxib, deracoxib, diclofenac, diflunisal, ethenzamide, etofenamate, etoricoxib, fenoprofen, flufenamic acid, flurbiprofen, lonazolaka, lornoxicam , ibuprofen, indomethacin, izoksikama, kebuzona, ketoprofen, ketorolac, naproxen, nabumetone, nifluminovoy acid, sulindac, tolmetin, piroxicam, meclofenamic acid, mefenamic acid, meloxicam, metamizol, mofebutazona, oxyphenbutazone, parecoxib, fenidona, phenylbutazone, piroxicam, propatsetamola, propiphenazone, rofecoxib, salicylamide, suprofen, thiaprofenic acid, tenoxicam, valdecoxib, 4- (4-cyclohexyl-2-methyloxazol-5-yl) -2-fluorobenzenesulfonamide, N- [2- (cyclohexyloxy) -4-nitrophenyl] 2- (3,4-difluorophenyl) -4- (3-hydroxy-3-methylbutoxy) -5- [4- (methylsulfonyl) phenyl] -3 (2H) -pyridazinone and 2- (3,5-difluoro phenyl) -3- [4- (methylsulfonyl) phenyl] -2-cyclopenten-1-one.

[00111] In some embodiments, the 5HT1B / 1D serotonin receptor agonist is selected from the group consisting of eletriptan, frovatriptan, naratriptan, risatriptan, sumatriptan and zolmitriptan.

[00112] In some embodiments of the invention, the N-methyl-D-aspartate antagonist is selected from the group consisting of amantadine, aptiganel, besonprodil, budipine, conantokine G, delucemine, dexanabinol, dextromethorphan, dextropropoxyphene, felbamate, fluorofelcibbenamide, ipenoxazone, kaitocephalin, ketamine, ketobemidone, lanicemin, lycostinel, midafotel, memantine, D-methadone, D-morphine, milnacipran, neramexane, orphenadrine, remacemide, sulfazocine, RPL-12495 metabolite) amino-5-chloro-1- (phosphonomethyl) -1H-benzimidazole-2-propanoic acid, 1-aminocyclopentanecarboxylic acid, [5- (aminomethyl) -2 - [[[(5S) -9-chloro-2,3, 6,7-tetrahydro-2,3-dioxo-1H-, 5H-pyrido [1,2,3-de] quinoxalin-5-yl] acetyl] amino] phenoxy] acetic acid, α-amino-2- (2 -phosphonoethyl) cyclohexane propanoic acid, α-amino-4- (phosphonomethyl) benzeneacetic acid, (3E) -2-amino-4- (phosphonomethyl) -3-heptenoic acid, 3 - [(1E) -2-carboxy-2- phenylethenyl] -4,6-dichloro-1H-indole-2-carb acid, 8-chloro-2,3-dihydropyridazino [4,5-b] quinoline-1,4-dione-5-oxide salt with 2-hydroxy-N, N, N-trimethylethanammonium, N '- [2- chloro-5- (methylthio) phenyl] -N-methyl-N- [3- (methylthio) phenyl] guanidine, N '- [2-chloro-5- (methylthio) phenyl] -N-methyl-N- [3 - [(R) -methylsulfinyl] phenyl] guanidine, 6-chloro-2,3,4,9-tetrahydro-9-methyl-2,3-dioxo-1H-indeno [1,2-b] pyrazin-9- acetic acid, 7-chlorothiokinurenic acid, (3S, 4aR, 6S, 8aR) -decahydro-6- (phosphonomethyl) -3-isoquinolinecarboxylic acid, (-) 6,7-dichloro-1,4-dihydro-5- [3 - (methoxymethyl) -5- (3-pyridinyl) -4-H-1,2,4-triazol-4-yl] -2,3-quinoxalindione, 4,6-dichloro-3 - [(E) - ( 2-oxo-1-phenyl-3-pyrrolidinylidene) methyl] -1H-indole-2-carboxylic acid, rel- (2R, 4S) -5,7-dichloro-1,2,3,4-tetrahydro-4- [[(phenylamino) carbonyl] amino] -2-quinolinecarboxylic acid, rel- (3R, 4S) -3,4-dihydro-3- [4-hydroxy-4- (phenylmethyl) -1-piperidinyl] -2H-1 -benzopyran-4,7-diol, 2 - [(2,3-dihydro-1H-inden-2-yl) amino] acetamide, 1,4-dihydro-6-methyl-5 - [(methylamino) methyl] - 7-nitro-2,3-quinoxalindione, [2- (8.9-dioxo-2, 6-diazabicyclo [5.2.0] non-1 (7) -en-2-yl) ethyl] phosphonic acid, (2R, 6S) -1,2,3,4,5,6-hexahydro-3 - [( 2S) -2-methoxypropyl] -6,11,11-trimethyl-2,6-methano-3-benzazocin-9-ol, 2-hydroxy-5 - [[(pentafluorophenyl) methyl] amino] benzoic acid, 1- [2- (4-hydroxyphenoxy) ethyl] -4 - [(4-methylphenyl) methyl] -4-piperidinol, 1- [4- (1H-imidazol-4-yl) -3-butynyl] -4- (phenylmethyl ) piperidine, 2-methyl-6-phenylethynyl) pyridine, 3- (phosphonomethyl) -L-phenylalanine and 3,6,7-tetrahydro-2,3-dioxo-N-phenyl-1H, 5H-pyrido [1,2 , 3-de] quinoxaline-5-acetamide.

[00113] In some embodiments, the anti-inflammatory compound is selected from the group consisting of aspirin, celecoxib, cortisone, deracoxib, diflunisal, ethericoxib, phenoprofen, ibuprofen, ketoprofen, naproxen, prednisolone, sulindac, tolmetin, meloxicam, pyroxicam phenylbutazone, rofecoxib, suprofen, valdecoxib, 4- (4-cyclohexyl-2-methyloxazol-5-yl) -2-fluorobenzenesulfonamide, N- [2- (cyclohexyloxy) -4-nitrophenyl] methanesulfonamide, 2- (3,4 -difluorophenyl) -4- (3-hydroxy-3-methylbutoxy) -5- [4- (methylsulfonyl) phenyl] -3 (2H) pyridazinone and 2- (3,5-difluorophenyl) -3- [4- (methylsulfonyl ) phenyl] -2-cyclopenten-1-one.

[00114] The disclosed invention has numerous aspects, illustrated by the following non-limiting examples.

EXAMPLES

Example 1

[00115] The studies described herein used extracellular multiposition electrophysiological monitoring of the rate of propagation of RDs and two-photon laser scanning microscopy to visualize, in real time, changes in the size of dendritic spines caused by RDs in hippocampal slices in vitro. The effect of the new functional partial agonist of the glycine site GLYX-13 NMDAR receptor on the threshold value and speed of RD propagation, and the real-time effects of RD on the dendritic spike morphology were investigated. GLYX-13 from time to time completely prevented the induction of RD by a local increase in the extracellular concentration of potassium, and stably slowed its propagation rate. The passage of the RD through the CA1 field of the hippocampus caused a rapid contraction of dendritic spines, which reversed after the restoration of neuronal depolarization. GLYX-13 improved the speed and degree of return of dendritic spines to their original sizes and positions after RD, which indicates that this drug and other substances that modulate NMDA receptor activity can protect synaptic compounds in the brain from possible damage resulting from repeated migraine attacks.

General methods

Medicines

[00116] All solutions for external use and patch pipette were prepared using deionized distilled water (resistance> 18 MΩ * cm ^-2 ; Milli-Q system). Chemicals for the manufacture of solutions for intracellular and intracellular use were purchased from Sigma-Aldrich (St. Louis, MO, USA). AlexaFluor 594 was purchased from Molecular Probes.

Slicing and extracellular registration

[00117] The experiments were performed on hippocampal slices of Sprague-Dawley® rats aged 14 to 21 days (Taconic Farms, Hudson, NY). Rats were deeply anesthetized with isoflurane, sacrificed, and the brain was quickly removed and placed in oxygenated (95% O ₂ - 5% CO ₂ ) ice-cooled artificial cerebrospinal fluid (ICJ) containing (in mmol / L): 126 NaCl, 2 5 KCl, 2.6 CaCl ₂ , 1.3 MgCl ₂ , 1.25 NaH ₂ PO ₄ , 26 NaHCO ₃ and 11 glucose. The brain was hemisectioned, the frontal lobes were cut off, and the individual hemispheres were glued using cyanoacrylate glue to a stage immersed in ice-cooled ICJ, continuously oxygenated during the preparation of the slices. 400-micron coronal sections including the hippocampus were cut using a vibrating microtome (Leica 1200s) and transferred to a mating valve chamber for incubation at room temperature for a minimum of 1 hour before being transferred to a Haas-type interface chamber (Haas-style interface chamber) for registration at 32 ° C. Sections were perfused with ICG (4 ml / min; ICG mmol / L: NaCl 126; KCl 3; NaH ₂ PO ₄ 1.25; MgCl 1.3; CaCl ₂ 2.5; NaHCO ₃ 26; glucose 10), saturated 95% O ₂ /5% CO ₂ before the start of the experiment, and all drugs were applied from the bath.

[00118] Extracellular recording was performed using a MultiClamp 700B instrument (Axon Instruments) with Clampex (v. 9), filtering at 1 kHz and sampling at 3 kHz. Low-resistance discharge electrodes (1-2 MΩ after ICSC filling) made of thin-walled borosilicate glass were inserted into the apical dendrite of the terminal field of the Schaffer collateral in the radiant layer of the CA1 field, at a distance of about 150 mm from the RD initiation pipette, to monitor the propagation of the RD. The registration chamber for immersed samples was mounted on a Zeiss Axioskop 2FS vertical microscope equipped with infrared differential interference contrast (DIC) optics. To visualize changes in the luminescence of the slice caused by RD, a 10x objective was used. Luminosity changes were recorded every 100 ms using a cooled CCD camera (CoolSNAP HQ), controlled by a PTI control system. Electrophysiological data were analyzed using the Clampfit 9 software. Visualization data was digitized and reconstructed using Image J (NIH) software.

[00119] A stainless steel bipolar stimulating electrode (FHC, Bowdoin, ME) was placed in the collateral commissure Schaffer fibers in the CA3 field, and a pulsed current was applied with stimulus intensity calculated to achieve approximately 50% of the maximum field excitatory postsynaptic potential of the pEPSP every every 30 s (from 50 to 100 pA; duration 100 μs). The electrical stimulation of the ISO-Flex isolator was monitored using an eight-pulse Master generator (AMPI, Jerusalem, Israel) and initiated using a Multiclamp 700V (Molecular Devices, Sunnyvale, CA). The signals were digitized using a Digidata 1322 and recorded using a Multiclamp 700V amplifier. The slope tangent of the pEPSP dependence was measured by linear interpolation in the region of 20-80% of the maximum inverse deviation, and the slope should remain stable within plus or minus 10% for at least 15 minutes before the start of the experiment. Data was analyzed using Clampfit (Version 9; Axon Instrument) on an IBM-compatible personal computer. The induced e-cess (50% of the maximum amplitude, 2-4 mV) was recorded in the field of the apical dendrite in the radiant layer for at least 30 minutes of a stable base period.

Hippocampal Depression Induction Induction

[00120] Acute coronal sections of the hippocampus were transferred to the registration chamber for immersed samples on a microscope stage and perfusion was performed with heated ICJ (32 ° C), and the level of extracellular [K ⁺ ] increased to 8 mmol / L at a perfusion rate of 3 ml / min. Propagating depression was initiated by extruding 3M KCl from a glass pipette. Pressure pulses of 55-69 kPa (8-10 psi) lasting 50-100 ms were generated using Picospritzer II (Parker Hannifin, Hollis, NH). The resistance values of the pipette filled with 3M KCl were 2 ± 0.2 MΩ.

Analysis of reduction / restoration of dendritic spine by dynamic imaging

[00121] After pumping the CA1 pyramidal neuron into a section of the Alexa Fluor 594 hippocampus (100 μmol / L) for 15 minutes, tertiary dendrites were visualized using a specially adapted Olympus BX61WI two-photon laser scanning microscope equipped with a 60x / 1.1 nA water-immersion infrared lens with 4x digital zoom as previously described (Zhang et al., 2008). We used the XYZ scanning mode in the range of +/- 3 μm from the focusing layer in order to avoid deviations during movement, with a step interval of 0.5 μm along the z axis, and each image was formed for 0.45 seconds, with a passage time of a single-row depth profile of 7 seconds . Increased precautions were taken in order to avoid both saturation of fluorescence and the appearance of any signs of phototoxicity of small dendritic spines from an excess of exciting light. Depth profile measurements were repeated at 5 min intervals to reduce possible photobleaching of the dye and phototoxicity. A Mai / Tai (Solid-State Laser, Mountain View, CA) laser tuned to a wavelength of 810 nm was used for excitation, and image acquisition was controlled using the Olympus FluoviewFV300 application program (Olympus America, Melville, NY). Epifluorescence was detected with the maximum open point diffraction of the photomultipliers of the confocal laser scanning head, and the emission spectral window was optimized with respect to the signal-background ratio. In the transfluorescence path, a 565 nm dichroic mirror was used to separate the green and red fluorescence, and passed through HQ525 / 50 and HQ605 / 50 emission filters, respectively, to eliminate transmitted or reflected excitation light (Chroma Technology, Rockingham, VT), and were detected simultaneously by two photomultipliers. The Figures show compressed images of the full 6 μm Z-profile, and these projections were used to calculate the intensity as an indicator of the spike volume.

Data analysis

[00122] The recorded signals were passed through an eight-pole Bessel low-pass filter with a cutoff frequency of 1 kHz and sampled using Clampex (V. 9) with an interval of 100 μs. After determining the values of the slope of the pESP using the Clampfit (V. 9), the data were further processed using the Origin 6.1 software (Microcal Software, MA) and presentations were prepared using CorelDraw 10 (Corel, Ottawa, Ontario, Canada). All data was analyzed by univariate analysis of variance (ANOVA) or by t-student test using the SPSS application program (SPSS Inc., Chicago, IL). The significance level was set equal to P <0.05. Data are presented as mean ± standard error of the mean (SEM) for all experiments.

results

Focal pervasive depression induced by high [K ⁺ ] in the CA1 field of hippocampal slices

[00123] In artificial cerebrospinal fluid (ICG) with a level of [K ⁺ ] о increased to 8 mmol / L at 32 ° С, a single local extrusion (100 ms) of 3M [K ⁺ ] о on the field in the CA1 field of the radiant layer ( Figure 1, top left) with high probability induced RD, which propagated with a propagation speed of 9 ± 5.4 mm / min (n = 16) throughout the CA1 field of the hippocampus. An RD characterized by a negative shift of the field potential (Figure 1, top right) showed a maximum shift of -8 ± 1.5 mV (n = 30) in the synaptic layer of the radiant layer of the hippocampal CA1 field, which usually lasted more than 45 seconds.

[00124] RDs were induced every 10 minutes to check whether the region of negative potential shift (Figure 1, upper right, shaded region), which is a measure of the effective magnitude and duration of K ⁺ conductivity in the extracellular space, would remain stable during repeated RDs. Although the areas of negative potential shift recorded from the initiating electrode were larger than that of the distal diverting electrode located at a distance of 600 μm (Figure 1, below), there were no significant differences in the amplitude or propagation speed of individual episodes of successively repeating RDs caused at intervals 10 minutes at any of the sites (Bonferroni multiple comparison criterion, P> 0.20). These results show that reproducible negative shifts in the RD potential can be stably repeatedly induced in the CA1 field of the hippocampus at least the first ten (times).

GLYX-13 inhibits the spread and increases the refractory period of the spreading hippocampal depression

[00125] To test the hypothesis that a new partial agonist for the NMDA receptor coagonistic glycine site, GLYX-13, can increase the threshold or prevent RD by regulating NMDA receptor activation in the physiological range and preventing over-activation, GLYX-13 was applied from the bath with 1, 10 or 50 μmol / L on hippocampal slices for 30 min until a short-term discharge of high [K +] (1 mmol / L in a patch pipette) into the radiant layer of the CA1 field and attempted to induce RD. The dose-response relationship was built on the basis of the time of the release of high [K +], first in native sections, and then after 30 minutes of application in the GLYX-13 bath in each of the three tested concentrations (1, 10, or 50 μmol / L), recording DC potential for detecting taxiways.

[00126] After at least three RDs in drug-free ICG were first called, GLYX-13 sections were perfused and continued to cause RDs every ten minutes. As shown in Figure 2, above, for 10 μmol / L GLYX-13, there were no significant differences between the baseline values for the regions of individual consecutive RD episodes at the initiation site (Bonferroni multiple comparison criterion, P> 0.20), which indicates that GLYX-13 does not affect RD initiation.

[00127] Next, GLYX-13 was tested to verify whether the ratio of the amplitudes of the taxiways in the near (proximal) and far (distal) regions of the registration changes with repeated taxiways. A slice perfusion of 10 μmol / L GLYX-13 was carried out and continued to cause RD every ten minutes (Figure 2, bottom). The Bonferroni multiple comparison criterion (P> 0.20) again showed the absence of significant differences between the regions of individual consecutive episodes of RD in the control sections. GLYX-13 did not affect the initiation of a negative shift in RD potential at the proximal recording site.

[00128] The taxiway amplitudes at the initiation site correlated with the areas of negative shift of the taxiway potential at the remote recording site (Figure 2, bottom). Thus, the ratio of proximal / distal areas without GLYX-13 and in its presence was comparable. As shown in Figure 2, below, the ratio of the areas of the remote site and the site of initiation decreased sharply in the presence of 10 μmol / L GLYX-13 when calling the sixth taxiway (6th taxiway). Two-way analysis of variance showed the presence of three significant effects: 1) the effects of the time of repeated RDs in the same section (F (1.12) = 5.17, p <0.05) for all experiments; 2) effects of drug treatment (F (1.12) = 5.07, p <0.05); and 3) the interaction of the effects of time and processing (F (1.12) = 30.91, p <0.01). These results show that repeated negative shifts in the RD potential change the amplitude ratio between the remote portion and the initiation portion; and that this decrease was markedly enhanced by GLYX-13. These results indicate that GLYX-13 can significantly inhibit the spread of RD in the brain, especially after multiple RD events.

[00129] In previous studies, it was demonstrated that GLYX-13 can limit the propagation of RDs by decreasing the amplitude of the negative shift of the field potential and slowing the conduction velocity, the magnitude of these effects being greatest after multiple RDs. As a result of shortening the intervals between initiation of RD, it was found that each RD in the control sections has an absolute refractory period of 3-4 minutes until the next successful RD (n = 7). 10 μmol / L GLYX-13 increased this refractory period to 5-6 minutes (n = 6). As shown in Figure 3, the taxiway can be successfully called five minutes after the previous taxiway in the control slice (control), but not called in the slice processed GLYX 13 (GLYX-13 30 ').

Measurement of RD distribution using characteristic changes in optical density caused by RD induced extracellular space volume shifts

[00130] RD causes a deep reduction in extracellular space, as fluid rushes into depolarized cells, they swell, and this can be seen by a change in characteristic optical density in vivo. Such changes in luminous intensity can also be easily detected in sections of the brain using differential interference contrast microscopy (DIC) in transmitted light. Figure 4A shows that the "aura" of the RD in the form of increased luminosity diverges from the initiating pipette and propagates along the slice, and can be used to calculate the conductivity of the RD (Figure 4B). The time dependence of the increased luminosity correlates well with the maximum negative shift in the RD potential when using the results of electrophysiological measurements. Thus, calculations of the time dependence of the change in luminescence on distance provide an accurate and convenient way to measure the conductivity of the RD.

[00131] In order to determine whether the repeated XRs would exhibit the same conductivity rate, measurements were made of the changes in luminosity at 5 points located at a distance of 150 μm, as schematically shown in the upper part of Figure 4A. One-way analysis of variance with repeated measurements showed that the RD maintains a stable transmission speed from P1 to P5 (F (5.35) = 2.42, p> 0.05) in an individual section. Thus, the passage speed from P1 to P5 can be used to test the stability of the RD in successive episodes. The average RD propagation rate was 9.07 ± 0.55 mm / min, with values ranging from 7.54 to 11.14 mm / min (n = 16).

[00132] To test whether a stable rate is maintained during repeated episodes of the taxiway, and whether GLYX-13 affects the conductivity of the taxiway, repeated measurement analysis of variance was used to analyze 6 consecutive taxiways, as shown in Figure 5. The analysis revealed significant differences between control and GLYX-13 treated sections (F (1.50) = 3.66, p <0.01). The RD sequence accounted for 38.5% of the total dispersion, which indicates that the rate of RD propagation slows significantly with an increase in the number of RDs (F (5.50) = 14.01, p <0.01). Although the effect of GLYX-13 on the propagation speed of RDs over all repeated RDs does not reach a significance level according to the results of two-way analysis of variance, there is a significant interaction between the number of RDs and GLYX-13 processing (F (5.50) = 2.53, P <0, 05), indicating that GLYX-13 (filled circles) significantly reduces the speed of taxiway propagation only to the seventh taxiway.

Pervasive depression causes a reversible reduction in the volume of dendritic spines of pyramidal CA1 neurons.

[00133] The anatomical microarchitecture of pyramidal neuron dendrites in the brain is strikingly labile in response to various stimuli. Depolarization, oxygen / glucose deficiency, and N-methyl-D-aspartate have been shown to cause in vitro contraction of dendritic spines of pyramidal neurons CA1 of the hippocampus. To study the response of spines on RD, single CA1 pyramidal neurons were filled with AlexaFluor-594 fluorescent dye and visualized the shape of the dendritic spike using a stack of consecutive sections along the z-axis (with a step of 0.2 μm, the total length of 5 μm) obtained by the 2-photon method laser scanning microscopy. Thirty minutes after the result of pumping AlexaFluror-594 into the CA1 pyramidal neurons in equilibrium distribution in the neuron, RD was initiated by short-term ejection of a high [K +] (1M in the patch pipette) into the radiant layer of the CA1 field of the hippocampus slices, where the axons of the Schaffer collateral contact apical dendrites of pyramidal neurons CA1. As shown in Figure 6A and 6B, the depolarization generated by the RD causes a significant reduction in spines, determined by the fluorescence amplitude of the folded z-stack, and the volume of the spike is completely restored 20-30 minutes after the first RD. This confirms that one of the consequences of RD-induced depolarization is a change in the morphology of the dendritic spine.

GLYX-13 improves dendritic spike recovery after spreading depression

[00134] Finally, the effects of GLYX-13 on the dynamic morphological response of dendritic spines on RD were studied. The ability of GLYX-13 to reduce the reduction of spines in response to RDs or to improve their recovery was investigated. Figure 6A illustrates the reduction of spines in response to two episodes of RD in a control pyramidal neuron, and Figure 6B shows the same process in the presence of 10 μmol / L GLYX-13. Analysis of variance with repeated measurements reaches a significance level (F (8.64) = 17.53, p <0.001) for the results of measurements of the fluorescence intensity of dendritic spines, such as those shown in the upper part of Figure 6C, and indicates that this significance is mainly associated with time periods after the taxiway (F (8.64) = 6.18, p <0.01), demonstrating that although GLYX-13 (filled circles) does not affect the contraction caused by the taxiway, it provides restoration of the size of the spike after the taxiway (F (8.64) = 2.81, p <0.05).

Example 2

[00135] Hippocampal Slice Preparation: Experiments were performed using Sprague-Dawley® rats (Taconic Farms) aged 14 to 21 days. Rats were deeply anesthetized with isoflurane, sacrificed, and the brain was quickly removed and placed in oxygenated (95% O ₂ - 5% CO ₂ ) ice-cooled artificial cerebrospinal fluid (ICJ) containing (in mmol / L): 126 NaCl, 2, 5 KCl, 2.6 CaCl ₂ , 1.3 MgCl ₂ , 1.25 NaH ₂ PO ₄ , 26 NaHCO ₃ and 11 glucose. The brain was hemisectioned, the frontal lobes were cut off, and the individual hemispheres were glued using cyanoacrylate glue to a stage immersed in ice-cooled ICJ, continuously oxygenated during the preparation of the slices. 400-μm thick coronal sections containing the hippocampus were excised using a vibrating microtome (Leica 1200s) and transferred to a mating valve chamber for incubation at room temperature for a minimum of 1 hour before the start of the experiment.

[00136] Induction of pervasive hippocampal depression: Acute coronal sections of the hippocampus were transferred to a recording chamber for immersed samples on a microscope stage and perfused with heated ICG (32 degrees Celsius), with extracellular [K +] increasing to 8 mmol / L at a perfusion rate of 3 ml / min Propagating depression was initiated by extruding 3M KCl from a glass pipette. Pressure pulses of 55-69 kPa (8-10 psi), usually with a duration of 50 ms, were created using a picospritzer. The resistance values of the pipette filled with 3M KCl were 2 ± 0.2 MΩ.

[00137] Electrophysiological registration methods: Extracellular registration was performed using a MultiClamp 700V instrument (Axon Instruments) with Clampex (v. 9), filtering at 1 kHz and sampling at 3 kHz. Low-resistance (1-2 MΩ after ICSC filling) discharge electrodes made of thin-walled borosilicate glass were inserted into the apical dendrites of the terminal field of the Schaffer collateral in the radiant layer of the CA1 field, at a distance of about 150 mm from the RD initiation pipette, to monitor the propagation of the RD. The registration chamber for immersed samples was mounted on a Zeiss Axioskop 2FS vertical microscope equipped with infrared differential interference contrast (DIC) optics. To visualize changes in the luminescence of the slice caused by RD, a 10x objective was used. Luminosity changes were recorded every 100 ms using a cooled CCD camera (CoolSNAP HQ), controlled by a PTI control system. Electrophysiological data were analyzed using the Clampfit 9 software. Imaging data was digitized and reconstructed using ImageJ (NIH) software. All experiments were carried out in accordance with the approved protocol, corresponding to the recommendations of the National Institute of Health.

[00138] The results of Example 1 were analyzed for women and adolescent patients. The experimental protocol was used to initiate the propagation of depolarization (RD) in the CA1 field of intracerebral hippocampal sections in vitro. To improve visual observation of the distance of the spreading depression from the site of initiation and the speed of propagation of the taxiway at a greater distance, the 10x lens in accordance with Example 1 was replaced with a 4x lens to increase the observation area to 1.6 mm The registration chamber was modified to perform section perfusion on both sides; more efficient perfusion not only provided sufficient oxygen in the brain section, but also helped maintain a stable extracellular environment, since each episode of spreading depression caused a sharp change in both extracellular and intracellular ion composition .

[00139] Two groups of 2 months ovariectomized female Sprague-Dawley® rats were included in these group studies. One group of ovariectomized rats received daily injections of estrogen for 7 days starting on the 7th day after surgery, while the other was injected only with an oil carrier as a control for 7 days before the start of the experiments.

[00140] Changes in luminescence after extrusion of 3M potassium were visually assigned to the failure groups (<100 mm), local distribution (> 100 mm; <800 mm); or complete distribution (> 800 mm). Attempts were made to induce RD in 39 slices from 8 rats treated with oil, and 35 slices from 7 rats treated with estrogen. Table 1 shows the results of determining the frequency of cases of RD caused by the same method in two groups. In the group receiving the oil, the failure rate was significantly higher than in the group receiving estrogen (chi-square test, P = 0.014), which indicates that estrogen alone increases the tendency of brain tissue to exhibit RD in focal depolarization.

[00141] Glow changes were used to evaluate the transfer rate of pervasive depression, as shown in Figure 7 below. Propagation rates were calculated for both localized and complete RDs induced by focal extrusion of 3M potassium by dividing the RD distance between the observation sites by the time required for the RD to travel the distance between these observation sites. As shown in Figure 7, the average propagation speed in slices of estrogen-treated rats was 0.121 ± 0.013 mm / s, which is significantly higher than the propagation speed in slices of rats treated with oil control (0.083 ± 0.005 mm / s, P <0.05, t- Student's test). These data show that estrogen plays a significant role in enhancing both the propensity and speed of RD in the hippocampus and possibly also in the neocortex.

[00142] Studies have been conducted on how estrogen affects the severity of RD by measuring the maximum distance of RD distribution from the initiation site. As shown in FIG. 8, the final propagation point along the Schaffer collateral path, approximately determined by averaging the distance of the point at which the minimum detectable changes in luminescence (> 5% of changes compared to the baseline) are observed from the initiation point, and the distance to the closely located observation point at which luminescence changes are undetectable, and measurements of this distance from the initiation point. As shown in Figure 8, RD in sections of estrogen-treated rats covered a significantly greater distance (0.594 ± 0.071 mm; n = 18) than RD in sections of rat-treated rats (0.394 ± 0.051 mm; n = 16, P <0.05, Student t-test). Thus, RD in estrogen-treated rats was not only easier to invoke, but also spread over long distances.

[00143] Then, 10 μmol / L GLYX-13 was added to the perfusate after inducing 2 RDs with an interval of 10 minutes, and continued to induce RDs every 10 minutes to determine if an effect on the speed and intensity of RDs was observed. Figure 9 shows typical differences in RD induced in the estrogen-treated rat section prior to administration of GLYX-13 and after 50 minutes of treatment with 10 μm GLYX-13 in the bath. Glow changes were measured at 10 observation points to evaluate the effects of GLYX-13 on the taxiway. Although RD was still easy to initiate, as shown in Figure 9B, Figure 9A clearly shows that luminescence changes associated with RD were slowed in the presence of GLYX-13. The paired Student's t-test showed that GLYX-13 significantly reduced the average speed of RD distribution from 6.56 ± 0.57 mm / min to 4.96 ± 0.28 mm / min (n = 5, P <0.01, paired t-student test). The same experiments were carried out in slices of the hippocampus receiving rat oil. Although GLYX-13 reduced the average speed of RD propagation from 5.09 ± 0.61 mm / min to 4.50 ± 0.39 mm / min (n = 5), this decrease did not reach statistical significance.

[00144] A two-way analysis of variance with repeated measurements was also carried out to study the effect of GLYX-13 on the speed of RD propagation in two groups. As shown in Figure 10, the first significant effect is observed in estrogen-treated rats before and after application of GLYX-13 (F (1.8) = 3, l; p <0.05); and the second significant effect arises as a result of preliminary administration of GLYX-13 when comparing two groups (F (1.8) = 4.2; p <0.05). These results show that GLYX-13 exerts a stronger effect in estrogen-treated rats, significantly slowing the rate of RD progression to the level of rat-treated rats.

Example 3

Ways

Animals

[00145] Adult males (2–3 months old) of Sprague-Dawley® rats (RD) were purchased from Harlan (USA). The rats were kept in Lucite cages with an aspen-shaped poplar wood chip litter, with a light-dark cycle of 12 hours: 12 hours (the light was turned on at 5 hours), and with free access to Purina® rat food (USA) and tap water during the studies . Induction of traumatic brain damage

[00146] Traumatic brain damage was induced with a single shock wave in a modification for rats according to the protocol of Goldstein et al. (Goldstein, LE, et al. (2012) "Chronic traumatic encephalopathy in blast-exposed military veterans and a blast neurotrauma mouse model," Science Translational Medicine 4: 134ra160.) Adult male Sprague-Dawley® rats were anesthetized with isoflurane, inserted into both the ear of the rat, earmolds, and exposed the rat with a single shock wave of about 290 kPa (42 PSI) of helium generated by piercing a 0.36 mm (0.014 inch) thick polyester film. Control animals were kept outside the shock wave radius. Then, the animals were dosed with rapastinel (3 mg / kg, intravenously (IV)) or 0.9% sterile saline vehicle (1 ml / kg, intravenously) 1 hour after exposure to the shock wave. Rats were first anesthetized with 3.5-4% isoflurane, then their ears were protected with 1.5 × 1.5 mm foam inserts (Pura-Fit, Moldex-Metric INC, Culver City, California). Rats were placed in thoracic rodent holders with access to the head (Stoelting, USA) to protect their bodies, allowing free movement of the head, with the placement of the ends of aluminum shock tubes (183 × 61 cm; L-3 Applied Technologies, USA) in 10 cm from the heads. Rats were exposed once to a helium shock wave of about 290 kPa (42 PSI) generated by piercing a 0.36 mm (0.014 in) thick polyester film. Control animals were kept outside the shock wave radius. The animals were given a dose of rapastinel (3 mg / kg, intravenously) or 0.9% sterile saline vehicle (1 ml / kg, intravenously) 1 hour after exposure to the shock wave. The latent state was recorded before exiting anesthesia (Figure 11A). The moment of regaining consciousness was determined by the manifestation of blinking and rectifying reflexes and normal ability to move (normal rhythmic gait, non-circular gait, full support of body weight, some manifestations of sniffing and research behavior). Animals were dosed with GLYX-13 (3 mg / kg, intravenously) or 0.9% sterile saline vehicle (1 ml / kg, intravenously) 1 hour after shock wave exposure. N = 4-6 per group; * P <0.05 (Figure 11A) analysis of variance.

[00147] Positive Emotional Learning (PEL) Figure 11B presents the results of a single 3-minute session of testing positive emotional learning (PEL) conducted 24 hours after exposure to the shock wave using an intersubjective plan. N = 4-6 per group. Fisher's posterior PLSD test was used to evaluate the results (rapastinel + TPGM / vehicle + TPGM).

[00148] A non-specific active contact game was conducted as previously described (Burgdorf, J., et al. (2011) "Positive emotional learning is regulated in the medial prefrontal cortex by GluN2B-containing NMDA receptors," Neuroscience 192: 515-523 ), and testing was performed 3 hours and 2 weeks after dosing. The experimenter initiated a nonspecific active contact game with his right hand. Animals participated for 3 min in a non-specific active contact game, which consisted of alternating 15 blocks of non-specific game and 15 seconds of lack of stimulation. High-frequency ultrasonic vocalizations (ultrasounds) were recorded and analyzed by sonogram using Avasoft SASlab Pro (Germany), as described previously (ibid.). Quantitative determination of frequency-modulated 50 kHz ultrasound recorded during each of the periods of lack of stimulation was performed to measure PEL. Before testing, animals were not accustomed to playing with stimulation.

results

[00149] As shown in Figure 11A, animals subjected to TPGM showed greater recovery times after anesthesia compared with control animals on keeping conditions [F (1, 13) = 41.7, P <0.05]. As shown in Figure 11B, rapastinel (3 mg / kg, intravenously) eliminated TPGM-induced inhibition of positive emotional learning [F (2.12) = 10.0, P <0.05; Fisher's a posteriori PLSD criterion rapastinel + TPGM / vehicle + TPGM; monitoring of conditions of storage / vehicle + TPGM vs., P <0.05]. In addition, the recovery time after anesthesia did not correlate significantly with the PEL indices [r (15) = - 0.14, P> 0.05]. In a test of positive emotional learning, a single dose of rapastinel (3 mg / kg, intravenously) completely restored the deficits in learning and emotions induced by TPHM. Thus, in this preclinical model, rapastinel was effective in treating the main cognitive and affective symptoms of TPGM, and rapastinel appears to have therapeutic potential for treating RTRD.

EQUIVALENTS

[00150] Although specific embodiments of the present invention have been described, the above description of the invention is illustrative and non-limiting. A large number of embodiments of the present invention will be apparent to those skilled in the art after reading this description. The full scope of the present invention should be determined by the claims, together with their full scope of equivalents, together with such options.

[00151] Unless otherwise indicated, all numerical values expressing the amounts of ingredients, reaction conditions, parameters, descriptive features, etc. used in the text of the description and claims, should be construed as modified in all cases by the term "about". Accordingly, unless otherwise indicated, the numerical parameters given in the text of the description and the attached claims are approximate values that can vary depending on the desired properties that must be achieved according to this description.

[00152] All publications and patents referred to in this document, including those listed below, are hereby incorporated by reference in full, as if it was specifically and individually indicated for each individual publication or patent that they are incorporated by reference. In case of conflict, this application shall prevail, including any definitions given therein.

Claims (24)

1. A method of treating a migraine selected from episodic migraine, chronic migraine, retinal migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, menstrual migraine, abdominal migraine, childhood periodic syndromes or cluster headache, in a patient in need thereof, comprising a stage wherein a pharmaceutically effective amount of a compound represented by the formula is administered to said patient:

 or a pharmaceutically acceptable salt thereof. 2. The method according to p. 1, characterized in that the compound is administered to the patient at a dose of from about 0.01 mg / kg to about 1000 mg / kg, or from about 1 mg / kg to about 500 mg / kg of the compound. 3. The method according to p. 1 or 2, characterized in that the migraine is a retinal migraine, ophthalmoplegic migraine, acephalgic migraine, menstrual migraine, periodic periodic syndromes of children or cluster headache. 4. The method according to any one of paragraphs. 1-3, characterized in that the migraine is a migraine with aura (classic migraine). 5. The method according to any one of paragraphs. 1-4, characterized in that the migraine is accompanied by allodynia. 6. The method according to any one of paragraphs. 1-5, including the introduction of from about 1 mg / kg to about 10 mg / kg, from about 10 mg / kg to about 250 mg / kg, from about 20 mg / kg to about 150 mg / kg, from about 30 mg / kg to about 125 mg / kg, from about 40 mg / kg to about 110 mg / kg, from about 50 mg / kg to about 100 mg / kg, from about 60 mg / kg to about 90 mg / kg, or from about 70 mg / kg to about 90 mg / kg of compound. 7. The method according to any one of paragraphs. 1-5, including the introduction of about 1 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 50 mg / kg, about 75 mg / kg, or about 100 mg / kg of the compound. 8. The method according to any one of paragraphs. 1-7, including the introduction of the indicated compounds twice a day, about once a day, once every 2 days, once every 3 days, once every 4 days, once every 5 days, about once a week, about once every two weeks, or about once a month. 9. A method of treating, suppressing and / or preventing cortical spreading depression in a patient in need thereof, comprising administering to said patient a pharmaceutically effective amount of a compound represented by the formula:

 or a pharmaceutically acceptable salt thereof. 10. A method of treating or alleviating long-term post-migraine complications in a patient in need thereof, comprising administering to said patient a pharmaceutically effective amount of a compound represented by the formula:

 or a pharmaceutically acceptable salt thereof. 11. The method according to p. 9 or 10, characterized in that the compound is administered to the patient at a dose of from about 0.01 mg / kg to about 1000 mg / kg, or from about 1 mg / kg to about 500 mg / kg of the compound. 12. The method according to p. 9 or 10, comprising the introduction of from about 1 to about 20 mg / kg, from about 2 to about 15 mg / kg; from about 5 to about 10 mg / kg, from about 10 mg / kg to about 250 mg / kg, from about 20 mg / kg to about 150 mg / kg, from about 30 mg / kg to about 125 mg / kg, about 40 mg / kg to about 110 mg / kg, from about 50 mg / kg to about 100 mg / kg, from about 60 mg / kg to about 90 mg / kg, or from about 70 mg / kg to about 90 mg / kg of compound. 13. The method according to p. 9 or 10, comprising administering about 1 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 10 mg / kg, about 21 mg / kg, about 25 mg / kg, about 30 mg / kg, about 50 mg / kg, about 70 mg / kg, or about 100 mg / kg of the compound. 14. The method according to any one of paragraphs. 9-13, including the administration of the compound twice a day, about once a day, every 2 days, every 3 days, once every 4 days, once every 5 days, about once a week, about once every two weeks , or about once a month. 15. The method according to any one of paragraphs. 1-14, further comprising administering the compound in conjunction with an opioid, antidepressant, antiepileptic agent, non-steroidal anti-inflammatory drug (NSAID), serotonin 5HT1B / 1D receptor agonist, N-methyl-D-aspartate antagonist, or anti-inflammatory compound. 16. The method according to any one of paragraphs. 1-15, characterized in that the patient is a person. 17. The method according to any one of paragraphs. 1-16, characterized in that the patient is a woman. 18. The method according to any one of paragraphs. 1-17, characterized in that the patient is a patient of childhood or adolescence.

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