KR20170018072A - Methods of treating or ameliorating migraine - Google Patents

Abstract

In certain embodiments, the present invention provides a method of treating migraine (e.g., epilepsy, chronic migraine, retinal migraine, ocular migraine migraine, migraine migraine, migraine headache disorder, menstrual migraine, Childhood periodic syndrome or cluster headache). In certain embodiments, the present invention is also directed to a method of treating or alleviating a long-term post-migraine episode in a patient by administering a peptide NMDAR partial agonist. In certain other embodiments, the invention provides a method of treating, inhibiting or preventing a disease or condition caused by inhibiting cortical spreading or inhibiting cortical spread in a patient in need thereof, comprising administering a peptide NMDAR partial agonist . For example, methods of treating epilepsy, traumatic brain injury, and / or stroke are provided herein.

Description

[0001] METHODS OF TREATING OR AMELIORATING MIGRAINE [0002]

Cross-reference to related application

This application claims the benefit of US Provisional Application No. 62 / 015,727, filed June 23, 2014, and U.S. Provisional Application No. 62 / 109,386, filed January 29, 2015, each of which is incorporated herein by reference in its entirety, To the benefit and priority of.

-del-Rio et al., Curr. Opin. Neurol., 17(3):289-93, 2004] 참조). SD는, 세포외 칼륨의 국소 증가 및 피질의 큰 구역에 걸쳐 느린 탈분극의 자가 전파하는 파를 생성하는 글루타메이트의 방출에 의해 촉발된, 뇌파 측정 활성의 느리게 전파하는 억제이다. 대략 15 내지 30%의 편두통 환자가 조짐 편두통(migraine with aura)을 경험한다. Migraine is a primary episodic headache pain disorder associated with a relapsing and fragile attack that is usually poorly controlled by existing drug therapy focusing on vascular trigger substances. The onset of a migraine attack is predominantly predated by flash scotia or migraine symptoms caused by the development of a spreading depression (SD) (Ayata, Headache, 50: 725-30, 2010; Eikerman Haerter et al., Curr. Neurol. Neurosci. Rep., 10: 167-73, 2010; And

-del-Rio et al., Curr. Opin. Neurol., 17 (3): 289-93, 2004). SD is a slow-spreading inhibition of EEG activity triggered by local increases in extracellular potassium and release of glutamate, which produces self-propagating waves of slow depolarization across large areas of the cortex. Approximately 15 to 30% of migraine patients experience migraine with aura.

The mammalian central nervous system (CNS) is the brain that contains the neuroactive peptides somatostatin, choleseitokinin, VIP, substance P, enkephalin, neuropeptide Y (NPY), neurotensin, TRH, CCK, And many neuroactive peptides that generate special signaling in the spinal cord. (See generally The Biochemical Basis of Neuropharmacology, Cooper, Bloom and Roth, 5th ed., Oxford University Press, New York, 1986). Careful explanations of the complex signaling pathways operating in the CNS have identified specific receptors that are regulated by these neuroactive peptides that present important therapeutic targets for the various disorders associated with the CNS.

The N-methyl-D-aspartate (NMDA) receptor (NMDAR) is a receptor involved in stroke-related brain cell death, convulsive disorders, and neurodegenerative disorders including learning and memory. NMDAR also plays a central role in regulating normal synaptic transmission, synaptic plasticity and excitatory cytotoxicity in the central nervous system. NMDAR is further involved in long-term potentiation (LTP). LTP is a sustained enhancement of neuronal connections underlying learning and memory (see Bliss and Collingridge, 1993, Nature 361: 31-39).

Two common classes of glutamate receptors are characterized by the central nervous system (CNS). These are G-protein coupled receptor families of signaling proteins, and metabotropic glutamate receptors belonging to ion-stimulating glutamate receptors (Muir and Lees, Stroke, 1995, 26, 503-513). Ion-stimulating classes are further subdivided into AMPA, kainate and NMDA receptor subtypes by selective ligands that activate them.

NMDA mediated molecular agonists and antagonist compounds have been developed for potential therapeutic applications. However, most of these are associated with very narrow therapeutic indices and unwanted side effects including hallucinations, ataxia, irrational behavior and considerable toxicity, all of which limit its effectiveness and / or safety.

Thus, there remains a need for improved treatment of migraine and other related disorders by compounds that provide increased efficacy and reduced undesirable side effects.

In certain embodiments, the present disclosure relates to a method of treating migraine comprising administering a pharmaceutically effective amount of a GLYX peptide to a patient in need thereof. In certain embodiments, the migraine is selected from the group consisting of episodic migraine, chronic migraine, retina migraine, ophthalmoplegic migraine, acephalgic migraine, migrainous disorder, menstrual migraine, Migraine, childhood periodic syndrome and / or cluster headache. In certain embodiments, the migraine headache is a non-migraine migraine (general migraine headache). In certain embodiments, the migraine headache is a headache migraine (classic migraine headache). In certain embodiments, the migraine head is accompanied by allodynia.

For example, the disclosed compounds are useful for the treatment of migraine (e.g., seizure, migraine, chronic migraine, retinal migraine, ocular migraine, migraine migraine, migraine headache, menstrual migraine, abdominal migraine, childhood periodic syndrome or cluster headache) It can interact with or regulate the action of the glycine site of NMDAR functionally for treatment.

In certain embodiments, the present disclosure is directed to a method of treating, inhibiting and / or preventing cortical spread inhibition (SD) comprising administering a pharmaceutically effective amount of a GLYX peptide to a patient in need thereof. In certain embodiments, the disclosure relates to treating or ameliorating a long-term post-migraine episode in a patient in need thereof, comprising administering to the patient a pharmaceutically effective amount of a GLYX peptide.

In certain embodiments, the GLYX peptide is a pharmaceutically acceptable salt thereof, or derivative thereof, having the following structure, or having NMDAR partial agonist activity:

.

.

In one aspect, the invention is directed to a method of treating migraine in a patient in need thereof, which method comprises administering to said patient a pharmaceutically effective amount of a compound represented by the formula: or a pharmaceutically acceptable salt thereof, Includes:

.

.

In another aspect, the invention is directed to a method of treating, inhibiting and / or preventing cortical diffusing inhibition in a patient in need thereof, which method comprises administering a therapeutically effective amount of a compound represented by the following formula ≪ / RTI > to said patient: < RTI ID = 0.0 >

.

.

In another embodiment, the invention is directed to a method of treating or alleviating a long-term post-migraine episode in a patient in need thereof, which method comprises administering a therapeutically effective amount of a compound represented by the following, or a pharmaceutically acceptable salt thereof, Comprising administering to a patient:

.

.

As described in more detail below, drugs that are effective when administered in the symptoms will be beneficial to the patient, for example, by allowing intervention in the early stages of migraine. It is contemplated that the methods described herein are applicable in the treatment of symptom migraine including administration of GLYX peptides in the context.

Figure 1 shows the lesion high [K ⁺ ] induced diffuse suppression (SD) in the field CA1 of the hippocampal slice, resulting in a change in brightness reflecting the dispersive wave of many depolarizations of neurons and glia.
Figure 2 shows the data for the SD that occurred in the brain slice; There was no significant difference between the reference regions of the individual sequential illustrations of SD at the initiation site (Bonferroni Multiple Comparison Test, P > 0.20), indicating that GLYX-13 did not alter the onset of SD .
Figure 3 shows data demonstrating that GLYX-13 increases the refractory period for SD initiation. SD could be successful at 5 minutes after the previous SD in the control slice (control), but could not occur at the slice treated by GLYX-13 (GLYX-13 30 ').
4A shows that the SD "signature" of increased brightness can be used to diffuse from the initiation pipette, propagate across the slice, and calculate the SD conduction velocity (FIG. 4B).
Figure 5 shows a one-way analysis of variance (ANOVA) by repeated measurements to analyze six subsequent SDs to test whether GLYX-13 affects the SD conduction velocity and whether the SD repeatedly maintains a stable rate ).
Figure 6A shows the visible contraction in response to two illustrations of SD in control pyramidal neurons, Figure 6B illustrates the same procedure in the presence of 10 [mu] M GLYX-13; Figure 6C shows that GLYX-13 rescues the recovery of visible size after SD.
Figure 7A shows the diffusion distance and Figure 7B shows the diffusion rate of diffusion inhibition in rats (estrogen treated and non-oil treated), indicating that the diffusion rate is faster compared to the oil treated rats .
Figure 8 shows that SD at the slice from the estrogen treated rats migrates longer than the oil treated rats.
Figure 9A shows that the brightness change associated with SD in rat experiments is delayed in the presence of GLYX-13; Figure 9B shows that SD is induced in the presence of GLYX-13.
Figure 10 shows the effect of GLYX-13 (F (1,8) = 3.1; p < 0.05) on SD propagation velocity across estrogen-treated rats before and after application of GLYX-13; And preliminary exposure of GLYX-13 (F (1,8) = 4.2; p < 0.05) between the two groups.
Figures 11A-11B show the relief of blast induced learning defects of rapastinel (3 mg / kg IV; 1 hour after blasting) in the PEL test 24 hours after blast. FIG. 11A shows the blast recovery time (potential for normal walking) data. FIG. 11B shows the results of a single 3-minute positive emotional learning (PEL) test session performed 24 hours after blasting using inter-subject design. Each group N = 4 to 6. * P <0.05 (FIG. 11A) ANOVA, or (FIG. 11B) of the Fisher PLSD post-test, Rapa styryl Ner + TBI vs. vehicle + TBI.

Migraine Classification

Migraine was first categorized first in 1988. The International Headache Society recently updated its classification of headaches in 2004. According to this classification, migraine is primarily a primary headache with tension-type headache and cluster headache

Migraine is classified into seven subclasses (some of which include additional subclasses):

Migraine, or "generalized migraine" includes migraine headaches that do not accompany symptoms.

Signs Migraine, or "traditional migraine", includes migraine headaches usually accompanied by signs. Less commonly, the symptoms can occur with or without a headache. Two other variants are familial hemiparesis syndrome and sporadic hemiparesis syndrome, in which a person is accompanied by movement disorders with an ocular migraine. If an ancestor has the same condition, it is called "familial", otherwise it is called "sporadic". Another variant is basilar-type migraine, where headaches and signs are not motor disorders but are accompanied by difficulties in speaking, world rotation, ringing in the ears, or many other brain-related symptoms. This type was initially thought to be due to seizures of the cranial base arteries, the arteries supplying the brainstem. Guidelines for the diagnosis of migraine headache can be found in, for example, Eriksen et al., European Journal of Neurology 11: 583-591, 2004 and International Classification of Headache Disorders, Second Edition -II). &Lt; / RTI &gt; Signs Subtypes of migraine include those described in ICHD-II, such as the usual signs with migraine headaches (IHS 1.2.1), common signs with non-migraine headaches (IHS 1.2.2), headache- Includes signs (IHS 1.2.3), familial hemiparesis involuntary migraine (IHS 1.2.4), sporadic hemiparesis involuntary migraine (IHS 1.2.5), and two donor migraine (IHS 1.2.6).

Childhood periodic syndrome, often a pioneer of migraine, includes periodic vomiting (sometimes a period of strong vomiting), abdominal migraine (accompanied by abdominal pain, common areas), and childhood benign paroxysmal vertigo (occasional dizziness).

Retinal migraine includes migraine headaches with visual impairment or even temporary blindness in one eye.

Complications of migraine describe rare or rare frequent, or migraine headaches and / or signs associated with seizures or brain lesions.

Possible migraine has some characteristics of migraine, but describes a condition in the absence of sufficient evidence to diagnose it with migraine (in the presence of concomitant drug overuse).

Chronic migraine is a complication of migraine, a headache that meets the diagnostic criteria for migraine headaches and occurs over a longer time interval. Specifically, more than 15 days / month for more than 3 months.

There are four possible stages of migraine, but not all of the stages are necessarily experienced: (1) global symptoms that occur in the hours or days before the headache; (2) an indication that the headache progresses immediately; (3) a pain stage also known as a headache stage; And (4) epigastric symptoms that are experienced after the end of the migraine attacks.

Bulbous or precursor symptoms occur in about 60% of people with migraine with a 2 to 2 day outbreak before the onset of pain or symptoms. The symptoms may include a wide variety of symptoms including altered mood, irritability, depression or bliss, fatigue, craving for certain foods, muscle stiffness (especially in the neck), constipation or diarrhea, and sensitivity to odor or noise have. This can occur in people with an ocular migraine or no ocular migraine.

Symptoms are a temporary lesional neurological phenomenon that occurs before or during a headache. This is gradual over several minutes and generally lasts less than 60 minutes. Symptoms can be visual, sensory or motor in nature, and many people experience more than one. Visual effects are most common; This occurs at less than 99% of cases and is not accompanied by sensory or motor effects in more than 50% of cases. Blindness usually consists of scintillation (a zone of partial change of vision that can interfere with the ability of a person to blink and read or drive). This usually begins around the visual field center and then looks like a wall of casting or sexuality And spread to the side by a zigzag line. Usually lines are black and white, but some people also see lines with colors. Some people lose some of their visions (known as anti-Semitic), while others experience blurring.

Sensory sign is the second most common type; These occur in 30% to 40% of people with symptoms. Usually a pin-and-needle feel starts from one side of the hand and arm and spreads from the same side to the nose. The lower limb usually occurs after the perception of the position is lost due to the perception of the position. Other symptoms of the indication stage may include speech or language impairment, rotation of the world, and less commonly, exercise problems. Exercise symptoms indicate that this is a hemiplegic migraine, and weakness usually lasts longer than one hour, unlike other symptoms.

Symptoms can also occur without a subsequent headache. Also known as silent migraine headache, migraine headache is relatively rare and involves signs and other symptoms without subsequent headache (ie, no pain stages).

During the pain phase, the headache is usually unilateral, puffy, and usually mild in intensity. This usually comes gradually, and is exacerbated by physical activity. However, in over 40% of cases, pain may be bilateral, and neck pain is often associated. Bilateral pain is especially common in people with no symptoms of migraine. Less commonly, pain can occur primarily in the back or upper part of the head. Pain usually lasts 4 to 72 hours in adults, but usually lasts less than an hour in young children. The frequency of attacks is variable, life span is very small or several weeks and average is about one month.

Pain is often accompanied by nausea, vomiting, sensitivity to light, sensitivity to sound, sensitivity to odor, fatigue and irritability. In migraine with craniofacial migraine, which has neurological symptoms associated with brain stem or neurological symptoms on both sides of the body, a common effect involves sensation, whim and confusion of the world's rotation. Zone occurs in almost 90% of people, and vomiting occurs in about 1/3. Other symptoms may include visual acuity, non-rigidity, diarrhea, frequent urination, pale or sweating. Tenderness or tenderness of the scalp can occur when neck stiffness occurs.

The effect of migraine can last for several days after the main headache is over; This is called migraine headache symptoms. Many report a sick feeling at the site of migraine, and some report a loss of thought for a few days after the headache has passed. The patient may feel tired, have headache, cognitive impairment, gastrointestinal symptoms, mood swings and weakness.

GLYX peptide

GLYX-13 is a newly developed, fast acting, long-lasting antidepressant with an unprecedented regulatory action on the activation of the N-methyl-D-aspartate glutamate receptor (NMDAR). This material, acting at the obligatory synergist glycine site at the NMDA receptor that needs to be activated, normalizes the activation of this important receptor, increasing it when it is very low and inhibiting it when it is very high. Through this action, GLYX-13 increases the induction of long-term fortification (LTP) while inhibiting LTD of synaptic strength and restores normal LTP from hippocampus slices from old animals.

As used herein, the term "GLYX peptide" is a peptide having the NMDAR glycine site partial agonist / antagonist activity. GLYX peptides can be obtained by well known recombinant or synthetic methods, such as those described in US Pat. Nos. 5,763,393 and 4,086,196 (incorporated herein by reference). In some embodiments, GLYX means a tetrapeptide having the amino acid sequence Thr-Pro-Pro-Thr, or L-threonyl-L-prolyl-L-prolyl-L-threonine amide.

For example, GLYX-13 means a compound shown as follows:

.

.

Polymorphs, homologs, hydrates, solvates, free bases and / or suitable salts of GLYX-13 such as, but not limited to, acetate salts are also contemplated. The peptide may be in the form of a cyclic or acyclic, as further described in US 5,763,393. In some embodiments, GLYX-13 analogs may include Thr or insertion of one or more moieties from the group of Pro, or deleted, for example, CH _2, deletion of the OH or NH ₂ moieties. In another embodiment, GLYX-13 may be optionally substituted by one or more halogen, (optionally substituted by halogen or amino) C ₁ -C ₃ alkyl, hydroxyl and / or amino. The glycine site partial agonists of NMDAR are disclosed in US 5,763,393, US 6,107,271, and Wood et al., NeuroReport, 19, 1059-1061, 2008, the entire contents of which are incorporated herein by reference.

It is contemplated that the peptides disclosed herein may include both natural and unnatural amino acids, for example, all natural amino acids (or derivatives thereof), all unnatural amino acids (or derivatives thereof), or mixtures of natural and unnatural amino acids Can be understood. For example, one, two, three or more amino acids in GLYX-13 may each independently have a D- or L- configuration.

GLYX-13 may act predominantly in NR2B containing NMDAR and may not exhibit the traditional side effects of known NMDAR modulators, such as CPC-101,606 and ketamine. In certain embodiments, essentially non-sedating anti-migraine or other therapeutic effect can be produced by GLYX-13 when administered to a subject in a therapeutically effective amount. In a much different embodiment, GLYX-13 may not have the potential for abuse (e.g., it may not be habit-forming).

In some embodiments, GLYX-13 can increase AMPA GluR1 serine-845 phosphorylation. In certain embodiments, the glycogen synthase kinase 3? (GSK-3?) May be activated by GLYX-13. In some cases, the level of beta -catenin may be changed after administration of GLYX-13.

In some embodiments, a composition comprising GLYX-13 or GLYX-13 is administered in combination with an i.v. In vivo potency and / or brain level concentration.

In addition, GLYX-13 can have a broad therapeutic index compared to glycine site antagonists such as L-701,324, or other glycine site antagonists with a narrow therapeutic index, thus providing a very narrow range of capacity between therapeutic effect and ataxia . For example, L-701,324 had an anticonvulsant effect on the capacity to produce ataxia (Bristow, et al, JPET 279: 492-501, 1996). Similarly, a series of Merz compounds had an anticonvulsant effect in the capacity to produce ataxia (Parsons, et al., JPET283: 1264-1275, 1997).

Way

In one aspect, the invention is directed to a method of treating migraine in a patient in need thereof, which method comprises administering to said patient a pharmaceutically effective amount of a compound represented by the formula: or a pharmaceutically acceptable salt thereof, Includes:

.

.

In certain embodiments, the compound is administered to a patient at a dose of about 0.01 mg / kg to about 1000 mg / kg or about 1 mg / kg to about 500 mg / kg of the compound.

In certain embodiments, the migraine headache is seizure-like migraine, chronic migraine, retinal migraine, ocular migraine migraine, migraine headache, migraine headache disorder, menstrual migraine, abdominal migraine, childhood periodic syndrome or cluster headache.

In certain embodiments, the migraine is a seizure type migraine, a chronic migraine, a retina migraine, an ophthalmoplegia migraine, an electroless migraine or a cluster headache.

In certain embodiments, the migraine headache is a headache migraine (classic migraine headache).

In certain embodiments, the migraine headache is a non-migraine migraine (general migraine headache).

In certain embodiments, the migraine head is accompanied by allodynia.

In certain embodiments, the method comprises administering a therapeutically effective amount of a compound of the invention in an amount of about 1 to 10 mg / kg, about 10 mg / kg to about 250 mg / kg, about 20 mg / kg to about 150 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 of the compound.

In certain embodiments, the method comprises administering to a subject in need of treatment a therapeutically effective amount of a compound of formula I in an amount of about 1 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / , About 30 mg / kg, about 50 mg / kg, about 70 mg / kg, or about 100 mg / kg of the compound.

In certain embodiments, the method comprises administering to a subject in need thereof at least one dose per day, at least once per day, at once per day, once per day, once per day, once per day, once per day, once per day, once per day, once per week, 1, or about once every 4 weeks.

In another aspect, the invention is directed to a method of treating, inhibiting or preventing a disease or condition caused by inhibiting cortical spreading or inhibiting cortical spread in a patient in need thereof, said method comprising administering a therapeutically effective amount of Comprising administering to the patient a compound or a pharmaceutically acceptable salt thereof that is displayed:

.

.

In another embodiment, the invention is directed to a method of treating or alleviating a long-term post-migraine episode in a patient in need thereof, which method comprises administering a therapeutically effective amount of a compound represented by the following, or a pharmaceutically acceptable salt thereof, Comprising administering to a patient:

.

.

In certain embodiments, the compound is administered to a patient at a dose of about 0.01 mg / kg to about 1000 mg / kg of compound. In certain embodiments, the compound is administered to a patient at a dose of about 1 mg / kg to about 500 mg / kg.

In certain embodiments, the method comprises administering a therapeutically effective amount of a compound selected from the group consisting of about 1 mg / kg to 10 mg / kg, about 1 mg / kg to 20 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 to about 100 mg / kg, about 60 mg / kg to about 90 mg / kg, or about 70 mg / kg to about 90 mg / kg of the compound.

In certain embodiments, the method comprises administering to a subject in need thereof a therapeutically effective amount of at least one of the following: about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 50 mg / kg, about 70 mg / Mg / kg, 15 mg / kg, 20 mg / kg or about 100 mg / kg of the compound.

In certain embodiments, the method comprises administering to a subject about two times per day, about once per day, once per day, once per day, once per day, once per day, once per day, once per day, once per week, Once or once a month for example.

In certain embodiments, the method further comprises the co-administration of an opioid, an antidepressant, an antiepileptic agent, a nonsteroidal anti-inflammatory drug (NSAID), a serotonin 5HT1B / 1D agonist, an N-methyl-D-aspartate antagonist, do.

NMDAR activation may promote or even require the development of SD in many experimental simulations. Thus, compounds that prevent overactivation of NMDAR can be an important new therapeutic agent to reduce or even prevent the onset of migraine attacks. For example, GLYX-13, described below, inhibits the generation and propagation of SD in the in vitro hippocampal slice. In certain experiments, GLYX-13 can completely prevent the induction of SD by local increase in extracellular potassium concentration and / or if it is not blocked, this can completely slow the rate of SD propagation. Furthermore, GLYX-13 can improve the restoration of dendritic visibility to its original size after SD. For example, methods for treating migraine in a patient on a prophylactic and / or acute basis are provided herein. Such administration can alleviate the severity (or, in some embodiments, halting) migraine attacks in the patient.

In certain embodiments, the patient is a human. Patients considered include female patients and / or adolescent patients.

Migraine headache, migraine headache disorder, menstrual migraine, abdominal migraine, childhood periodic syndrome, or cluster headache), or in the treatment of a migraine headache, for example, in migraine-resistant patients (e.g., seizure disorders, chronic migraine, chronic migraine, Methods for treating refractory migraine in a patient suffering from migraine without, or not responding to, appropriate treatment of at least one, or at least two, other compounds or therapeutic agents, are also provided herein. (E. G., Seizure type migraine, chronic migraine, chronic migraine headache) in a patient who is resistant to therapy, comprising the steps of: a) identifying the patient with a therapeutic tolerance, and b) administering an effective dose of GLYX- , Retinal migraine, ocular muscle palsy migraine, migraine migraine, migraine headache disorder, menstrual migraine, abdominal migraine, childhood periodic syndrome or cluster headache) are provided herein. In certain embodiments, the migraine is an ataxia migraine.

In one embodiment, there is provided a method of treating a migraine headache comprising administering an effective amount of a GLYX peptide, e. G., In a single unit dose, to a patient in need thereof for the manufacture of a medicament for the treatment or prevention of migraine (e. G., Seizure migraine, chronic migraine, retina migraine, Methods for acutely treating a migraine headache, migraine headache disorder, migraine headache disorder, menstrual migraine, abdominal migraine, childhood periodic syndrome or cluster headache headache are provided herein. For example, a method of treating migraine in a patient in need thereof in the onset of a migraine attack comprising administering an effective amount of GLYX-13 in an acute (e. G., Single dose) . Such a method may alleviate the patient of at least one symptom of migraine for less than about 2 weeks, less than 1 week, less than 1 day, or less than 1 hour (e.g., less than 15 minutes, less than 1.5 hours) after the administration. In some embodiments, the method may alleviate a patient having at least one symptom of migraine for at least about 1 day, at least 1 week, or more than 2 weeks after the administration. For example, a method is provided herein that includes administering an effective amount of a GLYX peptide to a patient suffering from migraine, wherein the patient is administered an effective amount of a GLYX peptide as compared to the same patient to which another agent for treating migraine is administered At least one symptom of migraine is substantially relieved substantially early after the first administration. One of ordinary skill in the art will appreciate that this method of acute administration may be beneficial in a hospital or outpatient setting. The methods described herein may also be useful during the treatment of allodynia occurring during an ocular migraine.

The method may also be used in the treatment of patients with depression, traumatic brain injury, epilepsy, or at risk of stroke. For example, in one embodiment, a method of treating a traumatic brain injury comprising administering an effective amount of a GLYX peptide, such as GLYX-13, is provided herein. In another embodiment, a method of treating epilepsy comprising administering an effective amount of a GLYX peptide, e. G., GLYX-13, is provided.

In addition to cardiovascular disease, patients with sign migraine may be at increased risk for other neurological and / or mental conditions and disorders. For example, coincidence of major migraine with major depression or suicide attempts has been shown to increase the risk of spontaneous seizures (Hesdorffer et al., Epilepsy Res. 75 (2-3): 220-223, 2007) . Other conditions associated with the sign migraine include a significantly higher marker of NO activity, an increased incidence of depression, and a genetically biomarker correlation to a stroke substantially greater than that of the general population (Etminan et al., &Lt; RTI ID = 0.0 & MJ 330 (7482): 63, 2005). Migraine sufferers having or at risk for any of these conditions may take medicines to treat or manage the disease and these medicines may interact negatively with currently used medications for the treatment of migraine headache. As described herein, some of these conditions are contraindicated for tryptic therapy (e.g., stroke and hydatidactate treatment). In addition, the FDA has concluded that in relation to serotonin syndrome, a life-threatening symptom that may occur when a tryptan is used with a serotonin reuptake inhibitor (SSRI) or a selective serotonin / norepinephrine reuptake inhibitor (SNRI) And published public health recommendations. Thus, the methods described herein may be useful in the treatment of patients with depression or those suffering from or at risk of stroke.

Dosage

The dosage of any composition of the disclosure will vary depending on the symptom, the age and weight of the patient, the nature and severity of the disorder to be treated or prevented, the route of administration, and the form of the composition. Any of the present agents may be administered in a single dose or in divided doses. Dosages for the compositions of this disclosure can be readily determined by techniques known to those skilled in the art or as taught herein. Generally, the compound is administered in a daily dose of, for example, 0.05 mg to 3000 mg (measured as a solid form), e.g., from about 10 mg to about 500 mg, or, for example, from about 1 to about 200 mg / A successful result can be obtained. The dose range includes, for example, 10 to 1000 mg (e.g., 50 to 800 mg). In some embodiments, the compound of Formula I is administered at a dose of 50, 100, 150, 200, 225, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, Lt; / RTI &gt; Alternatively, the dosage can be calculated using the patient &apos; s body weight. For example, the dose of a compound administered to a patient, or a pharmaceutical composition thereof, may range from 1 to 500 mg / kg (e.g., 5 to 250 mg / kg). In an exemplary non-limiting embodiment, the dose is in the range of 5 to 200 mg / kg (e.g., 1, 2, 2.5, 5, 10, 15, 20, 25, 30, 35, 40, 45, (For example, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 mg / kg). In an exemplary non-limiting embodiment, the dose may range from 1 to 15 mg / kg, 50 to 100 mg / kg, 60 to 90 mg / kg, or 70 to 80 mg / kg.

GLYX-13 can provide a high therapeutic index. For example, GLYX-13 may be administered at a dose of about 1 to about 10 mg / kg, about 10 to about 200 mg / kg, such as about 30 mg / kg, about 75 mg / kg, or about 100 mg / kg i.v. Or &lt; / RTI &gt; subcutaneous dose range. In some embodiments, no ataxia occurs at a dose of, for example, 500 mg / kg (i.v).

The therapeutically effective amount of GLYX peptide required for therapeutic use will vary depending upon the type of condition being treated, the duration of the desired treatment time, the age and condition of the patient, and ultimately determined by the attending physician. The desired dose may be administered in single effective doses for 2 weeks, 1 week, 6 weeks, 5 weeks, 4 weeks, 3 weeks, 2 weeks, or 1 day, or several doses administered at appropriate intervals, , &Lt; / RTI &gt; may be conveniently administered at least four sub-doses.

The effective dose or amount, and any possible effect on the timing of administration of the agent, may need to be ascertained for any particular composition of the present disclosure. This can be accomplished by routine experimentation as described herein using at least one group of animals (preferably at least five animals per group) or, if appropriate, in a human experiment. The effectiveness and the method of treatment or prevention of any of the present compositions can be assessed by assessing the effect of the administration by administering the composition, measuring one or more applicable indices, and comparing the post-treatment value of the index to the value of the same index prior to treatment .

The exact time and amount of administration of any particular composition that will produce the most effective treatment in a given patient will depend upon a variety of factors including the activity, pharmacokinetics and bioavailability of the composition, the physiological condition of the patient (age, sex, disease type and stage, Condition, responsiveness to a given dosage, and type of drug), route of administration, and the like. The guidelines set forth herein may be used to optimize treatment, for example, to determine the optimal time and / or amount of administration, which requires only routine experimentation with monitoring of the subject and adjustment of dosage and / or timing do.

As the subject is being treated, the patient &apos; s health can be monitored by measuring one or more relevant indices at a predetermined time during the treatment period. The compositions, amounts, times of administration, and treatments comprising the agents can be optimized according to the results of such monitoring. The patient can be periodically reassessed to determine the degree of improvement by measuring the same parameters. Adjustment of the amount (s) of the composition to be administered and possibly the time of administration may be made based on this reassessment.

The treatment may be initiated with a smaller dose lower than the optimal dose of the compound. Thereafter, the dosage can be increased in small increments until an optimal therapeutic effect is obtained.

Since the occurrence and duration of effects of different substances may be complementary, the use of the present compositions may reduce the dosage required for any individual substance contained in the composition.

The toxicity and therapeutic efficacy of the present compositions may be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example to determine LD50 and ED50.

Data from cell culture assays and animal studies can be used to formulate a range of dosage for use in humans. The dosage of any of the present compositions is preferably in the range of circulating concentrations that include ED50 with little or no toxicity. Dosages may vary within this range depending on the dosage form employed and the route of administration used. For compositions of this disclosure, the therapeutically effective dose can be initially predicted from cell culture assays.

In certain embodiments, the GLYX peptide is administered as a prophylactic measure (i. E., Prior to the pre-symptomatic phase). In certain embodiments, the GLYX peptide is administered during prodromal symptoms. In certain embodiments, the GLYX peptide is administered in an indication. By "on indication" is meant any time after onset of symptoms and before the onset of migraine pain. In certain embodiments, the GLYX peptide is administered after the onset of migraine pain. In certain embodiments, the GLYX peptide is administered during a glomeruloneal condition to, for example, reduce its symptoms.

The term "treat ", as used herein, refers to a prophylactic treatment or treatment to prevent one or more symptoms or conditions of the disease, disorder or condition described herein (e.g., pain or signs of migraine and allodynia Accompanied or absent). Prophylactic treatment may be initiated prior to (e. G., Pre-exposure prevention) or after (e. G., Post-exposure prevention) an event prior to the onset of a disease, disorder or condition (e. Prophylactic treatment, including administration of a GLYX peptide, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition thereof, as described herein, may be acute, short term or chronic. The dose administered may vary during the course of prophylactic treatment. See also Kaniecki et al., "Treatment of Primary Headache: Preventive Treatment of Migraine." In: Standards of Care for Headache Diagnosis and Treatment. Chicago (IL): National Headache Foundation; 2004, p. 40-52].

Formulation

The GLYX peptides of the present disclosure can be administered by various means depending on the intended use thereof as is well known in the art. For example, when the compositions of the present disclosure are administered orally, it can be formulated as tablets, capsules, granules, powders or syrups. Alternatively, the formulation of the present disclosure can be administered parenterally as an injection (intravenous, intramuscular, or subcutaneous), drop-on injection, or as a suppository. For application by ocular mucosal route, the compositions of the present disclosure may be formulated as eye drops or ointments. The formulation can be prepared by conventional means and, if desired, the composition can be used with any conventional additives such as excipients, binders, disintegrants, lubricants, correctives, solubilizers, suspending aids, emulsifiers or coatings .

DNA encoding a GLYX peptide, incorporated into an expression vector, may also be administered using any known method of administration to express the GLYX peptide in vivo.

In the formulations of the present invention, wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, and colorants, release agents, coatings, sweeteners, flavoring and perfuming agents, preservatives and antioxidants may be present as formulatory agents.

The compositions may be suitable for oral, topical (including buccal and sublingual), rectal, vaginal, aerosol and / or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. The amount of composition that can be combined with the carrier material to produce a single dose will vary depending upon the subject being treated, and the particular mode of administration.

The method of manufacture of this preparation comprises the step of placing the associative composition of the present disclosure with the carrier and optionally one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into liquid carriers, or finely divided solid carriers, or both, and associative materials, and then, if necessary, shaping the product.

Formulations suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a base, usually sucrose and acacia or tragacanth), powders, granules, or as solutions or suspensions in aqueous or non- , Or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base such as gelatin and glycerin, or sucrose and acacia) Of the present composition. The composition of the present disclosure may also be administered as a bolus, soft or paste.

In the case of solid dosage forms (capsules, tablets, pills, dragees, powders, granules, etc.) for oral administration, the compositions may contain one or more pharmaceutically acceptable carriers, such as sodium citrate or di-calcium phosphate, and / (1) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol and / or silicic acid; (2) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and / or acacia; (3) wetting agents such as glycerol; (4) disintegrators such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, specific silicates, and sodium carbonate; (5) solution retarders such as paraffin; (6) absorption accelerators such as quaternary ammonium compounds; (7) wetting agents such as acetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof; And (10) a colorant. In the case of capsules, tablets and pills, the composition may also comprise buffering agents. Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or lactose, and high molecular weight polyethylene glycols and the like.

Tablets may optionally be made by compression or molding with one or more accessory ingredients. Compressed tablets may be prepared by conventional means such as binding agents (e.g., gelatin or hydroxypropylmethylcellulose), lubricants, inert diluents, preservatives, disintegrants (e. G., Sodium starch glycolate or crosslinked sodium carboxymethylcellulose) May be prepared using an active or dispersing agent. Molded tablets may be prepared by molding a mixture of the present composition wetted by an inert liquid diluent in a suitable machine. Tablets, and other solid dosage forms, such as dragees, capsules, pills, and granules, may optionally be prepared by shading or other coatings well known in the art of coatings and shells such as enteric coatings and pharmaceutical formulations.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the present compositions, the liquid dosage forms may be formulated with inert diluents commonly used in the art such as water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate (Especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofuryl alcohol, sorbitol, Polyethylene glycols of fatty acids, fatty acid esters, cyclodextrins, and mixtures thereof.

Suspensions may contain, in addition to the present compositions, suspensions of the active ingredient, such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, , &Lt; / RTI &gt; and the like.

Formulations for rectal or vaginal administration may be prepared by mixing the compositions with one or more suitable non-excipient excipients or carriers, including, for example, cocoa butter, polyethylene glycol, suppository wax or salicylate, , But may be presented as a suppository that is liquid at body temperature and therefore melts in the body cavity and releases the active agent. Formulations suitable for vaginal administration also include parcels, tampons, creams, gels, pastes, foams or spray formulations containing carriers as are known in the art as being suitable.

Dosage forms for transdermal administration of the compositions include powders, sprays, ointments, pastes, creams, ointments, gels, solutions and patches.

For topical ocular administration, the compositions of the present invention may take the form of solutions, gels, ointments, suspensions or solid inserts in unit dosage amounts formulated to contain a therapeutically effective amount of the active ingredient, .

The pharmaceutical compositions of the present invention suitable for parenteral administration may be reconstituted with an aseptic injectable solution or dispersion immediately prior to use and may be formulated to contain an antioxidant, a buffering agent, a bacterial growth inhibiting agent, an agent that is isotonic with the blood of the intended recipient Dispersions, suspensions or emulsions, or sterile powders, which may contain one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, solubilizers or suspending agents or thickening agents.

Examples of suitable aqueous and nonaqueous carriers that may be used in the pharmaceutical compositions of the present invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), and mixtures thereof, vegetable oils , Such as olive oil and injectable organic esters such as ethyl oleate and cyclodextrin. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

Combination therapy

Any of the GLYX peptides described herein (e. G., GLYX-13) may be used alone or in combination with other substances to treat or prevent any of the diseases or conditions described herein. For example, in some combination therapies, one or more dosages of the therapeutic compound may be decreased from a standard dosage when administered alone.

The methods of the invention also include the concomitant administration of GLYX peptides with opioids, antidepressants, antiepileptics, non-steroidal anti-inflammatory drugs (NSAIDs), serotonin 5HT1B / 1D agonists, N-methyl-D-aspartate antagonists or anti-inflammatory compounds.

This disclosure is based, in some embodiments, on the use of single or more than one other antidepressant treatment such as tricyclic antidepressant, MAO-I, SSRI, and dual and triple reabsorption inhibitors and / or migraine A GLYX peptide in combination with an anxiolytic drug for the manufacture of a medicament for treating a migraine headache, a migraine headache, a retinal migraine headache, an ocular migraine headache, a migraine headache disorder, a migraine headache disorder, a menstrual migraine headache, Lt; / RTI &gt; peptide. Exemplary drugs that may be used in combination with the GLYX peptide include anapranil, adapine, avenyl, elabyl, And include rheumatoid, desirel (trazodone), and rudiodmil. In addition, the compounds of the present invention can be used in combination with other drugs.

In certain embodiments, the opioid is selected from the group consisting of alfentanil, butorphanol, buprenorphine, dextromodamide, dezocine, dextrorphoxifene, codeine, dihydrocodeine, diphenoxylate, , Hydrocodone, hydro morphone, ketobemidone, loperamide, levorphanol, levomadone, mettazinol, methadone, morphine, morphine-6-glucononide, nalbuphine, naloxone, oxycodone, oxymorphone, penta Is selected from the group consisting of benzoic acid, benzoic acid, benzoic acid, benzoic acid, benzoic acid, benzoic acid, benzoic acid, benzoic acid,

In certain embodiments, the antidepressant is selected from the group consisting of adinazolam, allproclaite, aminethytin, amitriptyline / chlorodiaxanthoxide combination, atipamozole, azamylancerin, vasinaprin, befuralin, bipemelan, , Biphenamol, broparomine, carpazone, sericramine, cianoframine, cymoxatone, citalopram, celefrole, clofammin, tadafinil, decanol, But are not limited to, ziprin, ditinepine, droxidopa, enecethexine, esthazolam, etoparidone, femetecine, fengabine, But are not limited to, but are not limited to, thiazolidinediones, thiazolidinediones, thiazolidinediones, thiazolidinediones, thiazolidinediones, Norfloxacin, orotrielin, oxaflozan, pinazepam, pirolindol, pizotillin, ri Wherein the compound is selected from the group consisting of serine, thiamine, threonine, threonine, threonine, serine, threonine, threonine, threonine, threonine, threonine, threonine, Tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol, tocopherol and tocopherol.

In certain embodiments, the anti-epileptic agent is selected from the group consisting of carbamazepine, flupirtine, gabapentin, ramotrygin, oxcarbazepine, phenytoin, retigavine, topiramate, and valproate.

In certain embodiments, the NSAID is selected from the group consisting of acetemethicone, aspirin, celecoxib, deracoxib, diclofenac, diflunisal, ethenamicide, etopenamate, etoricoxib, fenoprofen, flufenamic acid, Norepinephrine, norepinephrine, norepinephrine, norepinephrine, norepinephrine, norepinephrine, norepinephrine, norepinephrine, norepinephrine, norepinephrine, norepinephrine, But are not limited to, camphor, camphor, camphor, camphor, camphor, camphor, camphor, camphor, camphor, camphor, camphoric acid, (4-cyclohexyl-2-methyloxazol-5-yl) -2-fluorobenzenesulfonamide &lt; / RTI &gt; 4- (3-hydroxy-3-methylbutoxy) -4-nitrophenyl] methanesulfonamide, N- [2- (cyclohexyloxy) Phenyl] -3 (2H) -pyridazinone, and 2- (3,5-difluorophenyl) -3- [4- (methylsulfonyl) phenyl] 2-cyclopenten-l-one.

In certain embodiments, the serotonin 5HT1B / 1D agonist is selected from the group consisting of eletriptan, probutriptan, narutriptan, lizatriptan, sumatriptan and zolmitriptan.

In certain embodiments, the N-methyl-D-aspartate antagonist is selected from the group consisting of amantadine, attitergen, verson prodyl, todipine, conanthone G, delulcamine, dexanthinol, dextromethorphan, dextrope D-methadone, D-methadone, D-mannitol, D-mannitol, D-mannitol, D-mannitol, (Α) -α-amino-5-chloro-1- (phosgene), morphine, milnacipran, neramexane, orphenadrine, remasemide, sulfamazone, FPL-12,495 (racemate metabolite), topiramate (5S) -9-chloro-2,3,6,7-tetrahydro-naphthalen-2-ylmethyl) 5-yl] acetyl] amino] phenoxy] -acetic acid, a-amino-2 - (2-phosphonoethyl) -cyclohexanepropanoic acid, -amino 3-heptenoic acid, 3 - [(1E) -2-carboxy-2-phenylethenyl] Chloro-2,3-dihydropyridazino [4, 5-dichloro-lH-indole-2-carboxylic acid, 2-hydroxy-N, N, , 5-b] quinoline-1,4-dione 5-oxide salt and N '- [2-chloro-5- (methylthio) - [2 - [(R) -methylsulfinyl] phenyl] -guanidine, 6-chloro-2,3 (3S, 4aR, 6S, 6S) -7-methyl-2,3-dioxo-1H-indeno [1,2- b] pyrazine- (6,7-dichloro-1,4-dihydro-5- [3- (methoxymethyl) -5- (3-pyridinyl) -4-H-1,2,4-triazol-4-yl] -2,3-quinoxalindane, Oxo-1-phenyl-3-pyrrolidinylidene) methyl] -1H-indole Carboxylic acid, (2R, 4S) -rel-5,7-dichloro-1,2,3,4-tetrahydro-4 - [[(phenylamino) carbonyl] amino] -2-quinolinecarboxylic acid, Dihydro-3- [4-hydroxy-4- (phenylmethyl) -1-piperidinyl] -2H-1-benzopyran-4,7-diol Dihydro-6-methyl-5 - [(methylamino) methyl] -7-nitro (8,9-dioxo-2,6-diazabicyclo [5.2.0] non-1 (7) -en-2-yl) ethyl] -phosphate (2R, 6S) -1,2,3,4,5,6-hexahydro-3 - [(2S) -2-methoxypropyl] -6,11,11-trimethyl- 1- [2- (4-hydroxyphenoxy) ethyl] - methano-3-benzazocin-9-ol and 2-hydroxy-5 - [[(pentafluorophenyl) (4-methylphenyl) methyl] -4-piperidinol, 1- [4- (1H- 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.

In certain embodiments, the anti-inflammatory compound is selected from the group consisting of aspirin, celecoxib, cortisone, deracoxib, eracoxib, diflunisal, etoricoxib, fenoprofen, ibuprofen, ketoprofen, naproxen, (4-cyclohexyl-2-methyloxazol-5-yl) benzoic acid, lauric acid, linalk, tolmetin, piroxycam, mefenamic acid, meloxicam, phenylbutazone, rofecoxib, 2- (3,4-difluorophenyl) -4- (3-hydroxymethyl) -2-fluorobenzenesulfonamide, N- [2- (cyclohexyloxy) -3-methylbutoxy) -5- [4- (methylsulfonyl) phenyl] -3 (2H) -pyridazinone and 2- (3,5- difluorophenyl) -3- [4- Methylsulfonyl) phenyl] -2-cyclopenten-l-one.

The present disclosure has a number of aspects exemplified by the following non-limiting examples.

Example

Example 1

The studies described herein used SD velocities of radio waves and extracellular multi-site electrophysiologic monitoring of 2-photon laser scanning microscopy for real-time imaging of SD-induced changes in dendritic visibility size in in vitro hippocampal slices . The effect of the novel NMDAR receptor functional glycine site partial agonists of GLYX-13 was investigated in the real time effects of SD on the rate and threshold of propagation of SD and on the dendritic visibility form. GLYX-13 prevented the induction of SD from time to time by locally increasing the extracellular potassium concentration, and slowed its propagation rate constantly. The passage of SD through the hippocampal CA1 region produced a rapid contraction of the dendritic spines that reversed after neuronal depolarization was restored. GLYX-13 improves the rate and extent of the return of dendritic spines to the original size and position of the posterior segment of the SD, allowing the other to regulate drug and NMDA receptor activity from synaptic connections in the brain from possible damage from repeated migraine attacks Protection.

General method

drug

All external and patch pipette solutions were made with deionized distilled water (resistance greater than 18 MΩ cm ^-2 ; Milli-Q system). Chemicals for making extracellular and intracellular solutions were purchased from Sigma-Aldrich (St. Louis, MO, USA). Alexa Fluor 594 was purchased from Molecular Probes.

Slice preparation and extracellular recording

Experiments were performed on hippocampus slices from Sprague-Dawley (TM) rats (Taconic Farms, Hudson, NY) between 14 and 21 days of age. Anesthetic depth by a rat in iso-fluoran and, expense and, removing the brain quickly and, 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 (mM units (95% O ₂ -5% CO ₂ ), ice-cooled artificial cerebrospinal fluid (ACSF). The brains were bisected, the forehead cut, and the individual hemispheres were adhered using cyanoacrylate attached to the stage, which was subsequently immersed in oxygenated ice-cold ACSF during slicing. A 400 mu m thick slice slice containing hippocampus was cut using a Leica 1200s and transferred to an interface holding chamber for incubation at room temperature for a minimum of 1 hour and then transferred to a Haas style interface I moved to the chamber. The slices saturated with _{95% O 2/5% CO} 2 prior to the start of the experiment ACSF (4㎖ / min; ACSF mM: NaCl 126; KCl 3; NaH 2 PO 4 1.25; MgCl 1.3; CaCl 2 2.5; NaHCO 3 26; Glucose 10), and all drugs were bath-wetted.

Extracellular recordings were performed by Clampex (v. 9) using MultiClamp 700B (Axon Instruments), filtered at 1 kHz and digitized at 3 kHz. A low resistance (1 to 2 M OMEGA after being charged by ACSF) recording electrode was inserted into the apical dendritic zone of the Schaffer collateral termination field in the emissive layer of the CA1 zone at intervals of about 150 mm from the SD initiation pipette &Lt; / RTI &gt; was made from a thin walled borosilicate glass and the rate of SD was monitored. A recording chamber immersed in a Zeiss Axioskop 2 FS vertical microscope equipped with infrared differential interference contrast (DIC) optics was mounted. The 10x objective lens was used to image the slice luminance change caused by SD. The luminance change was imaged every 100 ms by a cooled CCD camera (CoolSNAP HQ) controlled by the PTI master system. Electrophysiological data were analyzed by Clampfit 9. Imaging data was digitized and reconstructed by ImageJ (NIH).

Place a bipolar stainless steel stimulation electrode (FHC (Main State Press)) in a Schaffer collateral-commissural fiber in the CA3 area and force each 50 seconds to produce approximately 50% of the maximum fEPSP every 30 seconds 100 pA; 100 기간 period). Current pulses were applied by the adjusted stimulus intensity. Electrical stimulation from an ISO-Flex isolator was controlled by a master eight pulse generator (AMPI, Israel Jerusalem) and triggered by Multiclamp 700B (Molecular Devices, Sunnyvale, Calif.). Signals were digitized by Digidata 1322 and recorded using a Multiclamp 700B amplifier. The fEPSP slope was measured by linear extrapolation from 20-80% of the maximal sound deflection and the slope was confirmed to be stable within 10% for at least 15 minutes before starting the experiment. Data was analyzed using Clampfit (Version 9; Axon Instrument) on IBM compatible personal computers. The induced fEPSP (50% of maximum amplitude, 2 to 4 mV) was recorded in the peak dendritic field in the emissive layer for a stable baseline period of at least 30 minutes.

Induction of hippocampal diffusion inhibition

The hippocampal acute parietal slice was transferred to a recording chamber immersed in a microscope stage and perfused with warmed ACSF (32 ° C) where the extracellular [K ⁺ ] was elevated to 8 mM at a perfusion rate of 3 ml / min. Diffusion inhibition was initiated by a pressure injection puff of 3M KCl through a glass pipette. A pressure pulse of 8-10 psi lasting 50-100 ms was driven by Picospritzer II (Parker Hannifin, Holliss, NH). The pipette resistance was 2 ± 0.2 MΩ when filled with 3M KCl.

Dendrite  Dynamic visualization analysis of visual contraction / recovery

CA1 pyramidal neurons were loaded in hippocampal slices by Alexa Fluor 594 (100 [mu] M) for 15 minutes, then customized two-photon laser-equipped with 60x / 1.1 nA water immersion infrared objective and 4x digital room as described above, Scanning A third dendrite projection was imaged using an Olympus BX61WI microscope (Zhang et al., 2008). An XYZ scanning mode in the range of +/- 3 mu m from the focused layer was used at z stage intervals of 0.5 mu m from the focused layer to avoid movement bias and each image took 0.45 seconds to the end for a single depth profile time of 7 seconds. I took a lot of attention to avoid random indications of phototoxicity and saturation of fluorescence for small dendritic spines from too much excitation light. Depth profiles were repeated at 5 minute intervals to reduce possible dye fade and phototoxicity. Mai / Tai lasers (Solid-State Laser, Mountain View, CA) adjusted to 810 nm were used here and image acquisition was controlled by the Olympus FluoviewFV300 software (Olympus America, Melville, NY). Simultaneous fluorescence was detected by a scanhead photomultiplier tube of a confocal laser with a maximally open pinhole and an emission-optimized spectrum window for the signal to background. In the transfluorescent path, green and red fluorescence were separated using a 565 nm color-picking mirror and passed through HQ525 / 50 and HQ605 / 50 emission filters, respectively, to transmit or reflected excitation light (Chroma Technology Locking ham)) were removed and detected simultaneously by two photomultiplier tubes. The figure shows a collapsed image of the entire 6 탆 Z-profile, which projection was used to calculate the intensity as an exponent of the visible volume.

Data Analysis

The recording signal was filtered by a 1 kHz cutoff frequency through an eight-pole Bessel low-pass filter and sampled by Clampex (V.9) at intervals of 100 μs. After the fEPSP slope was calculated by Clampfit (V.9), the data was further processed by Origin 6.1 (Microcal Software, MA) and presented by CorelDraw 10 (Corel, Ottawa, Ontario, Data were analyzed by Student's t test using one-way analysis of variance (ANOVA) or SPSS software (SPSS Inc., Chicago, IL) .The significance level was set at P &lt; ± SEM.

result

In the field CA1 of the seahorse slice Lesional  High [K +] induced Diffusibility  control

(Fig. 1, upper left) of 3M [K ⁺ ] o into the field in the CA1 emissive layer in artificial cerebrospinal fluid (ACSF) where [K ⁺ ] o rose to 8 mM at 32 [ SD was reliably induced, spreading across the entire CA1 region of the hippocampus at a propagation velocity of 9 ± 5.4 mm / min (n = 16). SD characterized by a negative field potential shift (Fig. 1, upper right) showed a maximum shift of -8 ± 1.5 mV (n = 30) in the radial layer synaptic layer of the hippocampus CA1 region, which typically lasted more than 45 seconds .

SD was induced every 10 minutes to test whether the area of the negative potential transfer (Fig. 1, upper right, hatched area), which is a measure of the period of extracellular space conduction and the effective scale of K ⁺ , was stable by repeated SD. The area of the voice potential shift recorded from the start electrode is 600 占 퐉 far higher than in the distal recording electrode (Fig. 1, lower), but the amplitude or propagation speed of individual illustrations of repeated sequential SD There was no difference (multiple comparison of this ferroni, P > 0.20). This result indicates that the reproducible SD speech potential shift can occur stably and repeatedly in the field CA1 of the hippocampus for at least the first ten.

GLYX-13 inhibits propagation and increases the refractory period of hippocampal spread inhibition

To test the hypothesis that the novel NMDA receptor glycine co-agonist site partial agonist GLYX-13 can raise or prevent the limit on SD by regulating NMDA receptor activation within the physiological range and preventing excessive activation, Prior to the simple injection of high [K +] (1 mM in the patch pipette) into the emissive layer of μ, GLYX-13 was applied to the hippocampal slice at 1, 10, or 50 μm for 30 min and attempted to induce SD. The dose-response relationship was initially plotted on the basis of the time of high [K +] injection on the Naïve slice, and after 30 minutes thereafter, each of the three test concentrations (1, 10 or 50 μM) Was applied with a bath of GLYX-13.

At least three SDs were induced initially in drug-free ACSF, and then the slice was perfused with GLYX-13 and continued to induce SD every 10 minutes. In the case of GLYX-13 at 10 [mu] M as illustrated in Figure 2 above, there was no significant difference between the reference regions of the individual sequential illustrations of SD at the initiation site (this ferroni multiple comparison test, P > 0.20) 13 does not change the start of SD.

GLYX-13 was then tested to see if it affected the SD amplitude relationship at the near (proximal) and distant (distal) recording sites by repeated SD. The slice was perfused with 10 [mu] M GLYX-13 and SD was continuously induced every 10 minutes (Fig. 2, lower). This Peroni multiple comparison test ( P > 0.20) again did not show any significant differences between the zones of the individual sequential illustrations of SD in the control slides. GLYX-13 did not affect the initiation of the SD negative-potential translation at the proximal recording site.

The amplitude of SD at the initiation site was correlated with the region of the negative potential site of SD at the remote site (Fig. 2, bottom). Therefore, the ratio of proximal / distal area in the absence and presence of GLYX-13 was compared. As illustrated in Figure 2, lower, the area ratio between the remote site and the initiation site decreased dramatically in the presence of 10 μM GLYX-13 by application of the 6 th SD (6 th SD). Two-way ANOVA showed three significant effects: 1) Time effect repeat SD (F (1, 12) = 5.17, p <0.05) on the same slice compared to all observations; 2) Effect from medication (F (1,12) = 5.07, p <0.05); And 3) interaction of treatment with time (F (1,12) = 30.91, p <0.01). This result suggests that recurrent negative displacement of SD changes the amplitude ratio between the remote site and the initiation site; Indicating that this reduction is markedly enhanced by GLYX-13. This finding suggests that GLYX-13 can significantly inhibit the spread of SD through the brain, especially after multiple SD events.

Previous studies have demonstrated that GLYX-13 can limit the propagation of SD by reducing the amplitude of the speech field potential shift and slowing the conduction rate, the greatest effect after multiple SDs. By shortening the interval between each SD initiation, it was found that each SD in the control slice had an absolute refractory period of 3-4 minutes during the next successful SD (n = 7). 10 μM GLYX-13 prolonged this refusal period by 5-6 min (n = 6). As shown in FIG. 3, SD could successfully occur at 5 minutes after the previous SD in the control slice (control), but could not occur at the slice treated by GLYX-13 (GLYX-13 30 ').

SD  Judo Extracellular  Using the intrinsic variation of the optical density produced by the space displacement SD  Measurement of radio wave

As the fluid rushes into the depolarized cells and it swells, SD causes profound contraction of extracellular space volume, which can be seen as a change in intrinsic optical density in vivo. This intensity intensity change can also be easily detected in brain slices using propagated optical DIC microscopy. 4A shows that the SD "signature" of increased brightness can be used to diffuse from the initiation pipette, propagate across the slice, and calculate the SD conduction velocity (FIG. 4B). The time course of the increased luminance is well correlated with the maximum voice potential shift of SD using electrophysiological recordings. Thus, the calculation of the time course of the luminance change by distance provides an accurate and convenient way to measure the SD conduction velocity.

To determine if repeated SDs exhibited the same conduction rate, the luminance change was monitored over five points separated by 150 [mu] m as programmed at the top of Fig. 4A. One-way ANOVA by repeated measurements shows that SD maintains the normal permeation rate within the individual slice from P1 to P5 (F (5, 35) = 2.42, p> 0.05). Thus, the permeation rate can be used from P1 to P5 to test SD stability over successive illustrations and to test over successive illustrations. The average propagation velocity of SD is 9.07 0.55 mm / min, and is in the range of 7.54-11.14 mm / min (n = 16).

To test whether the repeated illustrations of SD maintain stable rates and whether GLYX-13 affects SD conduction velocity, the ANOVA test is used by repeated measurements to analyze six subsequent SDs as illustrated in Figure 5 Respectively. The analysis showed a significant difference between the control and the GLYX-13 treated slice (F (1,50) = 3.66, p <0.01). The SD sequence accounts for 38.5% of the total variance, indicating that the SD propagation velocity is slowed by the significantly increasing number of SDs (F (5,50) = 14.01, p <0.01). There was a significant interaction between the number of SDs and the GLYX-13 treatment (F (5,50) = 2.53), even though the GLYX-13 effect on SD propagation rate across all repeats SD did not reach significance in the two-way ANOVA , P <0.05), reflecting that GLYX-13 (filled circle) significantly reduces the SD propagation rate only by the 7th SD.

Diffusibility  Inhibition is the CA1 pyramidal neuron Dendrite  Allow thorns to shrink reversibly in volume

The anatomic microstructure of the pyramidal neuron resin cells in the brain is surprisingly reliable in response to various stimuli. Depolarization, oxygen / glucose deprivation and N-methyl-D-aspartate all produced shrinkage of dendritic spines in CA1 pyramidal neurons in hippocampal hippocampus. To investigate the response of visibility to SD, a single CA1 pyramidal neuron was filled with fluorescent dye Alexa Fluor-594, and a serial z-stack slice (0.2 탆 step spanning 5 탆) collected from a 2-photon laser scanning microscopy examination Was used to image the thorn morphology. After 30 minutes of loading of the Alexa fluoro-594 into the CA1 pyramid reached equilibrium distribution in the neurons, SD was elevated to the radial layer of the CA1 region of the hippocampus slice (capillary side axon synapses in CA1 pyramidal neurons) K +] (1M on a patch pipette). As illustrated in Figures 6A and 6B, the depolarization produced by SD does not cause substantial visible shrinkage as measured by the collapsed z-stack fluorescence amplitude, and the visual volume is 20-30 minutes after the first SD . This confirms that one of the consequences of SD-induced depolarization is a deformation of the dendritic spine morphology.

GLYX -13 is Diffusibility  After inhibition Dendrite  Thorn recovery Improve

Finally, the effect of GLYX-13 on the dynamic morphological response of dendritic spines to SD was examined. It was investigated whether GLYX-13 responded to SD to reduce visual contraction or to improve its recovery. Figure 6A illustrates the visible contraction in response to two illustrations of SD in control pyramidal neurons, while Figure 6B illustrates the same procedure in the presence of 10 [mu] M GLYX-13. ANOVA for repeated measurements reached significance (F (8,64) = 17.53, p < 0.001) for the fluorescent intensity measured from the dendrite visibility as illustrated at the top of Figure 6C, (F (8,64) = 6.18, p < 0.01), indicating that GLYX-13 (filled circle) does not alter the shrinkage caused by SD, but this rescues the recovery of visible size after SD F (8,64) = 2.81, p < 0.05).

Example 2

Preparation of hippocampus slices: Experiments were performed using Sprague-Dawley (TM) rats (Taconic Farms) between 14 and 21 days of age. Anesthetic depth by a rat in iso-fluoran and, expense and, removing the brain quickly and, 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 (mM units (95% O ₂ -5% CO ₂ ), ice-cooled artificial cerebrospinal fluid (ACSF). The brains were bisected, the forehead cut, and the individual hemispheres were adhered using cyanoacrylate attached to the stage, which was subsequently immersed in oxygenated ice-cold ACSF during slicing. A 400 mu m thick slice slice containing hippocampus was cut using a Leica 1200s and transferred to the interfacial retention chamber for incubation at room temperature for at least 1 hour before starting the experiment.

Induction of hippocampal diffusibility inhibition: The hippocampal acute parietal slice was transferred to a recording chamber immersed in a microscope stage and perfused with warmed ACSF (32 ° C), where extracellular [K +] was perfused at a perfusion rate of 3 ml / min To 8 mM. Diffusion inhibition was initiated by a pressure injection puff of 3M KCl through a glass pipette. Pressure pulses of 8-10 psi, which lasted for 50 ms, were driven by a picospritzer. The pipette resistance was 2 ± 0.2 MΩ when filled with 3M KCl.

Electrophysiological recording method: Extracellular recordings were performed by Clampex (v. 9) using MultiClamp 700B (Axon Instruments), filtered at 1 kHz and digitized at 3 kHz. A low resistance (1 to 2 M OMEGA after being charged by ACSF) recording electrode was applied to the apical dendritic region of the capillary side end field in the emissive layer of the CA1 region at approximately 150 mm distance from the SD initiation pipette to the thin walled borosilicate Made from glass, the rate of SD was monitored. A recording chamber immersed in a Zeiss Axioskop 2 FS vertical microscope equipped with infrared differential interference contrast (DIC) optics was mounted. The 10x objective lens was used to image the slice luminance change caused by SD. The luminance change was imaged every 100 ms by a cooled CCD camera (CoolSNAP HQ) controlled by the PTI master system. Electrophysiological data were analyzed by Clampfit 9. Imaging data was digitized and reconstructed by ImageJ (NIH). All experiments were performed under approved protocols in accordance with the National Institutes of Health guidelines.

The findings of Example 1 have been investigated in connection with female and adolescent patients. Experimental protocols were used to initiate diffuse depolarization (SD) in the CA1 region of the hippocampus in vitro brain slices. To better observe the distance of diffusion suppression and the SD propagation velocity from the initiation site over a longer range of visibility, the 10x objective of Example 1 was replaced by a 4x objective to magnify the viewing domain to 1.6 mm . The recording chamber is modified so that the slice is perfused from both sides and that more effective perfusion not only provides sufficient oxygen to the brain slice but also allows each illustration of diffusing inhibition to produce dramatic ion composition changes both extracellularly and intracellularly Lt; RTI ID = 0.0 &gt; extracellular &lt; / RTI &gt; environment.

Two groups of 2 month old ovariectomized Sprague-Dawley (R) female rats were included in the current cohort study. One group of ovariectomized rats was provided with daily injections of estrogen for 7 days after the 7th surgical day while the other was only injected with oil vehicle as the control group for 7 days before the experiment.

Change in luminance was failed (<100 mm), local diffusion (> 100 mm; <800 mm); Or &lt; / RTI &gt; complete diffusion (&gt; 800 mm). SD induction was attempted on 39 slices from 8 oil-treated rats and 35 slices from 7 estrogen-treated rats. Table 1 summarizes the frequency of occurrence of SD caused by the same method in the two groups. In the oil-treated group, the failure rate was significantly higher than in the estrogen-treated group (chi-square test, P = 0.014), indicating that estrogen itself increases the tendency of brain tissue to exhibit SD in lesion depolarization.

The luminance change was used to predict the propagation speed of the diffusion suppression as shown in FIG. The propagation velocity was calculated from both the localization and full SD induced by the lesion 3M potassium puff by dividing the distance of SD between the sites of observation by the time it took for the SD to travel between these sites. As illustrated in Figure 7, the average diffusion rate at the slice from the estrogen treated rats was significantly greater than the diffusion rate at the slice from the oil treated rats (0.083 0.005 mm / sec, P < 0.05, Student t test) Fast 0.121 ± 0.013 mm / sec. This data indicates that estrogens play a significant role in increasing both the tendency and speed of SD in the hippocampus, and possibly also in the neocortex.

A test was conducted to determine how estrogen affects the severity of SD by measuring the maximum distance of SD propagation far from the onset site. As shown in FIG. 8, the distance of the observation region showing the minimum detectable brightness change (> 5% change from the reference value) from the start position was averaged by the distance of the near neighbor observation region showing the undetectable luminance change, Lt; / RTI &gt; along the predicted path of the sharper side by measuring this distance from the diffusion endpoint. As shown in Figure 8, the SD at the slice from the estrogen treated rats was significantly higher than the SD at the slice from the oil treated rats (0.394 0.051 mm; n = 16, P <0.05, Student t test) The long distance (0.594 ± 0.071 mm; n = 18) was moved. Therefore, SD in estrogen-treated rats was not only more easily induced, but also propagated longer distances.

10 μM of GLYX-13 was then added to the perfusate after two Sd's were induced 10 minutes apart and continued to induce SD every 10 minutes to determine if the rate and scale of SD were affected. Figure 9 shows the typical difference in slice-induced SD from estrogen-treated rats before application of GLYX-13 and after 50 min exposure to 10 μM bath-applied GLYX-13. To evaluate the effect of GLYX-13 on SD, the change in luminance at 10 observation sites was measured. Although SD is still easily induced as shown in FIG. 9B, FIG. 9A clearly shows that the brightness change associated with SD is delayed in the presence of GLYX-13. The twin-t test indicates that GLYX-13 significantly reduces the mean SD propagation velocity from 6.56 ± 0.57 mm / min to 4.96 ± 0.28 mm / min (n = 5, P <0.01, paired t test). The same experiment was performed on hippocampal slices from oil-treated rats. This decrease did not reach statistical significance while GLYX-13 reduced the mean SD propagation velocity from 5.09 ± 0.61 mm / min to 4.50 ± 0.39 mm / min (n = 5).

Two-way repeated ANOVA was also performed to investigate the effect of GLYX-13 on SD propagation velocities across the two groups. As illustrated in Figure 10, the first significance arises from estrogen-treated rats (F (1,8) = 3.1; p < 0.05) before and after the application of GLYX-13; The second significance occurred between the exposure of GLYX-13 between the two groups (F (1,8) = 4.2; p < 0.05). These results indicate that GLYX-13 has a stronger effect in estrogen-treated rats, which significantly slows the spreading rate of SD to the level of oil-treated rats.

Example  3

Way

animal

Adult male (2-3 month old) Sprague-Dawley (SD) rats were purchased from Harlan (USA). The rats were housed in a Lucite cage that had white wood bedding and kept in 12 hours: 12 hours light: cancer cycle (light at 5AM) and were free to study Purina (R) rat food (US) Access was provided

Induce traumatic brain injury

Induced blunt-traumatic brain injury and modified for use in rats according to protocols such as Goldstein et al. (Goldstein, LE, et al. (2012) "Chronic traumatic encephalopathy in blast-exposed military veterans and a male adult Sprague-Dawley (TM) rats were anesthetized with isoflurane, the earplugs were inserted into both ears of the rats, and rats were anesthetized with polyester film &lt; RTI ID = 0.0 &gt;Lt; RTI ID = 0.0 &gt; PSI &lt; / RTI &gt; Siam controls were placed outside the blast radius. The rats were initially injected with 3.5-4% isoflurane (1 mg / kg IV), while the animals were dosed with 1 hour of rabastin (3 mg / kg IV) or 0.9% sterile saline vehicle And their ears were then protected by 1.5 x 1.5 mm foam plugs (Pura-Fit ear plugs, Moldex-Metric INC, Coolvertshire, Calif.). The rats were placed in a thoracic restrainer (Stoelting (USA)) to protect their bodies while allowing the head to move freely, and the head was placed in an aluminum shock tube (183 x 61 cm; L-3 Applied Technologies USA). &Lt; / RTI &gt; The rats received a single approximately 42 PSI blast of helium produced by 0.014 inch punching of the polyester film. Siam controls were placed outside the blast radius. Animals were dosed with either rafastinol (3 mg / kg IV) or 0.9% sterile saline vehicle (1 ml / kg IV) at 1 hour after blast. The potential for recovery from anesthesia was noted (Figure 11A). Recovery is defined as blinking and positional reflexes and evidence of normal gait (evidence of normal rhythm gating, acyclic gating, full weight gain, slight nasal congestion, and inquisitive behavior). Animals were dosed with GLYX-13 (3 mg / kg IV) or 0.9% sterile saline vehicle (1 ml / kg IV) at 1 hour after blast. N = 4-6 per group; * P &lt; 0.05 (FIG. 11A) ANOVA.

Positive Emotional Learning ( PEL ) Figure 11B shows the results of a single 3-minute positive emotional learning (PEL) test session performed 24 hours after blast using inter-subject design. N = 4-6 per group. Fisher PLSD post-test was used to evaluate the results (Rapastinel + TBI vs. Vehicle + TBI).

A heterogeneous rough-and-tumble play has been described by 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 the test occurred at 3 and 2 weeks after dosing. The heterogeneous coarse body play stimuli were administered by the right hand of the experimenter. The animals received a three-minute union coarse body play consisting of 15 seconds of alternating union coarse body play and 15 seconds of unenchanted play. A high frequency of ultrasound (USV) was recorded and analyzed by Sonogram as previously described (Id.) By Avasoft SASlab Pro (Germany). The frequency-modulated 50-kHz USV generated during each non-stimulation period was quantified to measure the PEL. Animals were not allowed to play stimuli before testing.

result

As shown in FIG. 11A, animals receiving TBI showed a longer recovery time from anesthesia as compared to Siam control animals [F (1, 13) = 41.7, P <0.05]. As shown in Figure 11B, rapastinel (3 mg / kg IV) relieved TBI induced inhibition of positive emotional learning [F (2, 12) = 10.0, P < Fisher PLSD post-test Rupastatin + TBI vs. Vehicle + TBI, Sham vs Vehicle + TBI vs. P <0.05]. In addition, the anesthetic recovery time was not significantly correlated with the rate of PEL [r (15) = -0.14, P > .05]. In the positive emotional learning test, a single dose of rapamystel (3 mg / kg IV) completely relieved the learning and emotion deficiencies induced by TBI. Thus, in this preclinical model, Rapastelnell is effective in treating major cognitions, and emotional symptoms of TBI and Rapastinel have therapeutic potential in the treatment of PTSD.

Equalization

Although specific embodiments of the present disclosure have been described, the foregoing description is illustrative and not restrictive. Many modifications of the present disclosure will be apparent to those skilled in the art upon examination of this specification. The full scope of the disclosure should be determined by reference to the claims and their full scope of equivalents, together with the specification and such modifications.

Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, parameters, technical characteristics and the like used in the specification and claims, and the like are to be understood as being modified in all instances by the term "about. &Quot; Thus, unless indicated to the contrary, the numerical parameters set forth in the specification and the appended claims are approximations that may vary depending upon the desired properties sought to be obtained by this disclosure.

All publications and patents mentioned in this specification, including the items listed below, are hereby incorporated by reference in their entirety, as if each individual publication or patent was specifically and individually indicated by reference. In the case of a conflict, the present application, including any definitions herein, will prevail.

Claims (26)

A method of treating migraine in a patient in need of treatment for migraine comprising administering to said patient a pharmaceutically effective amount of a compound as shown below or a pharmaceutically acceptable salt thereof.

2. The method of claim 1 wherein the compound is administered to the patient at a dose of about 0.01 mg / kg to about 1000 mg / kg or about 1 mg / kg to about 500 mg / kg of the compound. 3. The method of claim 1 or 2, wherein the migraine is selected from the group consisting of episodic migraine, chronic migraine, retina migraine, ophthalmoplegic migraine, acephalgic migraine, migraine disorder, , Menstrual migraine, abdominal migraine, childhood periodic syndrome, or cluster headache. The method according to claim 1 or 2, wherein the migraine headache is seizure type migraine, chronic migraine headache, retinal migraine headache, migraine headache, migraine headache, or migraine headache. 5. The method according to any one of claims 1 to 4, wherein the migraine is migraine with aura (classical migraine). 5. The method according to any one of claims 1 to 4, wherein the migraine is a non-migraine migraine (general migraine). 7. The method according to any one of claims 1 to 6, wherein the migraine is accompanied by allodynia. 8. The pharmaceutical composition according to any one of claims 1 to 7, which comprises about 1 mg / kg to about 10 mg / kg, about 10 mg / kg to about 250 mg / kg, about 20 mg / Kg to about 100 mg / kg, from about 60 mg / kg to about 90 mg / kg, from about 30 mg / kg to about 125 mg / kg, from about 40 mg / kg to about 110 mg / , Or from about 70 mg / kg to about 90 mg / kg. 8. A pharmaceutical composition according to any one of claims 1 to 7, which comprises about 1 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 10 mg / kg, about 20 mg / / Kg, about 30 mg / kg, about 50 mg / kg, about 75 mg / kg or about 100 mg / kg. 10. The compound according to any one of claims 1 to 9, wherein said compound is administered twice a day, once a day, once every two days, once every three days, once every four days, once every five days, About once, about once every two weeks or about once a month. A method of treating, inhibiting and / or preventing cortical spreading inhibition in a patient in need of such treatment, inhibition and / or prevention of a cortical spreading depression comprising administering a therapeutically effective amount of a compound represented by the formula A method for treating, inhibiting and / or preventing cortical diffusibility inhibition, comprising administering to said patient a pharmaceutically acceptable salt thereof:

. A method of treating or alleviating a long-term migraine after-effect in a patient in need of treatment or alleviation of a long-term migraine headache, comprising administering to said patient a pharmaceutically effective amount of a compound represented by the following formula or a pharmaceutically acceptable salt thereof: A method of treating or alleviating a long-term post-migraine episode, comprising:

. 13. The method of claim 11 or 12, wherein said compound is administered to said patient at a dose of about 0.01 mg / kg to about 1000 mg / kg or about 1 mg / kg to about 500 mg / kg of said compound. 13. The composition of claim 11 or 12, wherein said composition comprises about 1 to about 20 mg / kg, about 2 to about 15 mg / kg of said compound; About 10 mg / kg to about 250 mg / kg, about 20 mg / kg to about 150 mg / kg, about 30 mg / kg to about 125 mg / kg, about 40 mg / Kg to about 90 mg / kg, or from about 70 mg / kg to about 90 mg / kg. Way. 13. The method of claim 11 or 12 wherein said compound is administered at a dose of about 1 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 10 mg / kg, about 21 mg / 30 mg / kg, about 50 mg / kg, about 70 mg / kg or about 10 mg / kg. 16. The method according to any one of claims 11 to 15, wherein the compound is administered twice a day, once a day, once every two days, once every three days, once every four days, once every five days, About once, about once every two weeks or about once a month. 17. The pharmaceutical composition according to any one of claims 1 to 16, which is selected from the group consisting of opioids, antidepressants, antiepileptics, non-steroidal anti-inflammatory drugs (NSAID), serotonin 5HT1B / 1D agonists, Lt; RTI ID = 0.0 &gt; antagonist, &lt; / RTI &gt; or a phytate antagonist or anti-inflammatory compound. 18. The method according to any one of claims 1 to 17, wherein the patient is a human. 19. The method according to any one of claims 1 to 18, wherein the patient is female. 20. The method of any one of claims 1 to 19, wherein the patient is a pediatric or adolescent patient. A method of treating or alleviating traumatic brain injury in a patient in need of such treatment or alleviation of traumatic brain injury comprising administering to said patient a pharmaceutically effective amount of a compound represented by the following formula or a pharmaceutically acceptable salt thereof: How to treat or relieve traumatic brain injury:

. 22. The method of claim 21, wherein said compound is administered to said patient at a dose of about 0.01 mg / kg to about 1000 mg / kg or about 1 mg / kg to about 500 mg / kg of said compound, How to mitigate. 23. The method of claim 21 or 22, wherein said compound is administered in an amount of about 1 to about 20 mg / kg, about 2 to about 15 mg / kg; About 10 mg / kg to about 250 mg / kg, about 20 mg / kg to about 150 mg / kg, about 30 mg / kg to about 125 mg / kg, about 40 mg / Kg to about 90 mg / kg, or from about 70 mg / kg to about 90 mg / kg. A method of treating or alleviating traumatic brain injury. 24. The compound according to any one of claims 21 to 23, wherein the compound is administered twice a day, about once a day, once every two days, once every three days, once every four days, once every five days, About once, about once every two weeks, or about once a month. 25. A pharmaceutical composition according to any one of claims 21 to 24 for use in the manufacture of a medicament for the treatment and / or prophylaxis of steroids, antidepressants, opioids, antiepileptics, antiepileptics, anticonvulsants, NSAIDs, serotonin 5HT1B / 1D agonists, diuretics and / Or a pharmaceutically acceptable salt thereof. 26. The method according to any one of claims 21 to 25, wherein the patient is a human being, treating or alleviating traumatic brain injury.

Images