JP2005530778A - Methods for prevention and / or treatment of neurological disorders - Google Patents

Abstract

The present invention relates to the use of a substrate for UDP-glucuronosyltransferase (UGT) and salts thereof for the prevention and / or treatment of neurological disorders. In a preferred embodiment, the present invention relates to the use of at least one UDP-glucuronosyltransferase (UGT) substrate and salts thereof expressed in the brain for the prevention and / or treatment of neurological disorders. The invention further relates to the use of said substrates and salts thereof for the prevention and / or treatment of glutamate cytotoxicity, more particularly neuropathy induced by glutamate. The invention further relates to the use of said substrate and salts thereof for the manufacture of a medicament having an inhibitory effect on the release of glutamate.

Description

  The present invention relates to the use of a substrate for UDP-glucuronosyltransferase (UGT) and salts thereof for the prevention and / or treatment of neurological disorders. In a preferred embodiment, the present invention relates to the use of at least one UDP-glucuronosyltransferase (UGT) substrate and salts thereof expressed in the brain for the prevention and / or treatment of neurological disorders. The invention further relates to the use of said substrates and salts thereof for the prevention and / or treatment of glutamate cytotoxicity, more particularly neuropathy induced by glutamate. The invention further relates to the use of said substrate and salts thereof for the manufacture of a medicament having an inhibitory effect on the release of glutamate.

Numerous studies have established that cellular communication using excitatory amino acids may be converted into a mechanism of cell destruction.
For example, glutamate is a major excitatory neurotransmitter in the mammalian nervous system, particularly the brain and spinal cord, and operates at its diverse excitatory synapses.

  The wide distribution of glutamate receptors throughout the nervous system demonstrates that glutamate plays a central role in a wide range of physiological and pathological events (Watkins J. C., Collingridge GL ,, The NMDA receptor, IRL Oxford, 1989). For example, it has been strongly suggested that it plays a central role in functions such as learning, pattern recognition, and memory (Blist TVP, Collingridge GL, Nature 361, 31-39, 1993).

  Normally, extracellular glutamate levels rise in a spatially confined state for a short period with normal synaptic transmission; however, in pathological situations, the levels may remain significantly elevated.

  In addition, glutamate is toxic to neurons in vitro and in vivo, and the function of the glutamate receptor, particularly the N-methyl-D-aspartate ("NMDA") receptor subtype of glutamate receptor is numerous in neurons. It is also known from decades ago that it is critical in injury and nerve injury (Appel SH, Trends Neurosci. 16, 3-5, 1993). Many neurological disorders with epileptic seizures and chronic or acute degenerative processes such as Alzheimer's disease, Huntington's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), spinal muscular atrophy ( SMA), retinal disorders, strokes, depression and traumatic brain disorders are associated with neuronal cell death by glutamate receptor overstimulation. Similarly, in the ischemic brain, extracellular glutamate increases rapidly after the onset of ischemia and decreases with reperfusion, so that nerve damage caused by ischemia after cerebral artery occlusion is at least partially glutamate-receptive. It has also been shown that it can be mediated by body overactivation (Davalos et al., 1997, Stroke, 28, 708-710). Another pathological situation with a significant increase in extracellular glutamate concentration is hypoxia or hypoglycemia. Furthermore, Stephans and Yamamoto (1994, Synapse, 17, 203-209) have shown that the neurotoxic effects of drug-induced neurotoxicity, such as methamphetamine (METH), on striatal dopaminergic neurons are actually overstimulated by glutamate receptors. It can be mediated by

  Overactivation of these glutamate receptors (referred to as “glutamate cytotoxicity”) is actually associated with increased extracellular glutamate concentrations. Mechanisms of extracellular glutamate increase include increased glutamate excretion by cells and / or decreased glutamate uptake. Accordingly, providing a means of protecting diseased cells, particularly neurons, from glutamate-induced cytotoxicity, and more specifically providing a means of regulating glutamate release and / or uptake by glutamate producing cells. Would be desirable.

  To this end, glutamate receptors present in target cells, most often N-methyl-D-aspartate (“NMDA”) receptors, are targeted by inhibition with agonist or antagonist specific molecules. That has already been advocated. Examples of such molecules are anthranilic acid derivatives (see US 5,789,444), Basilen Blue D-3G (Reactive Blue 2) and Cibacron Blue 3GA. And 5-adenylylimidodiphosphate (AMPPNP) (see US 6,326,370), NMDA specific antagonists such as ketamine, gluconate (Sakaguchi et al., 1999, Neuroscience, 92, 677-684). ), Dextromophane, or 3- (2-carboxypiperazin-4-yl) -propyl-1-phosphonic acid (Kristensen et al., 1992, Pain, 51:24). -253; Eide et al., 1995, Pain, 61, 221-228), or 2-methyl-6- (phenylethynyl) pyridine (MPEP), an antagonist of metabotropic glutamate receptor subtype 5 (mGluR5) (Ossosska et al., 2001, Neuropharmacology, 41, 413-420).

  However, adverse side effects of these compounds (eg psychotic effects, headaches, hallucinations, discomfort, or impaired cognitive and motor functions) hinder their widespread use. Thus, currently available treatment methods are unsatisfactory in terms of safety or effectiveness for their widespread implementation.

  Therefore, there is still a need to provide improved methods and means for protecting diseased cells, more preferably neurons, from glutamate-induced cytotoxicity.

  Many neurological disorders are associated with increased levels of release of nerve mediators (or neurotransmitters) such as acetylcholine (Ach), butyrylcholine (Buch), γ-aminobutyric acid (GABA), serotonin, or catecholamines, particularly dopamine, or ATP. Related.

In mammals, glucuronidation can be used to remove many lipophilic drugs, pollutants, xenobiotics and endogenous compounds with less bioavailability and therefore less active, higher polarity and water solubility. It is the main metabolic pathway to promote detoxification and elimination by converting to conjugates and facilitating their transport to the excretory organ and subsequent excretion into the urine or bile (for review see Ritter , 2000, Chemico-Biological Interactions, 129, 171-193). This reaction is catalyzed by UDP-glucuronosyltransferase (UGT). This multigene family of enzymes links the glucuronic acid part of UDP-glucuronic acid via hydroxyl (alcoholic, phenolic), carboxyl, sulfuryl, carbonyl and amino (primary, secondary or tertiary) linkages. To structurally unrelated compounds (for review see Turkey and Strassburg, 2000, Annu. Rev. Pharmacol. Toxicol., 40, 581-616). Initially, glucuronidation was thought to be a metabolic pathway primarily performed in the liver. However, numerous studies have shown that UGT activity is also present in human intestines, kidneys, colon tissue and brain, in which the enzyme helps protect the organ against potentially reactive hydroxylated molecules that can reach the organ. It is thought to be actively involved (Suleman et al., 1998, Archives of Biochemistry and Biophysics, 358, 1, 63-67; Gradinaru et al., 1999, Neurochemical Research, 24, 995-1000).
US 5,789,444 US 6,326,370 Watkins J.M. C. , Collingridge G. L. The NMDA receptor, IRL Oxford, 1989. Bliss T.W. V. P. , Collingridge G. L. , Nature 361, 31-39, 1993. Appel S.M. H. , Trends Neurosci. 16, 3-5, 1993 Davalos et al. 1997, Stroke, 28, 708-710. Stephans and Yamamoto (1994, Synapse, 17, 203-209) Sakaguchi et al. 1999, Neuroscience, 92, 677-684. Kristensen et al. , 1992, Pain, 51: 249-253. Eide et al. , 1995, Pain, 61, 221-228. Ososska et al. , 2001, Neuropharmacology, 41, 413-420. Ritter, 2000, Chemico-Biological Interactions, 129, 171-193 Turkey and Strassburg, 2000, Annu. Rev. Pharmacol. Toxicol. , 40, 581-616 Suleman et al. , 1998, Archives of Biochemistry and Biophysics, 358, 1, 63-67. Gradinaru et al. , 1999, Neurochemical Research, 24, 995-1000.

  The inventors have otherwise utilized UGT activity in vivo, more specifically in the brain, for the treatment and / or prevention of acute and chronic nerve mediator related diseases or conditions, particularly neurological diseases. We have now demonstrated that we can identify a new class of compounds. The present invention is a pharmacological method that replaces conventional methods using compounds such as competitive and non-competitive glutamate antagonists or agonists, gangliosides and growth factors. In certain embodiments, the present invention can be used as a pharmacological tool to regulate neuronal mediator release and cytotoxicity, preferably neurotoxicity, by cells, many neurological disorders associated with epileptic seizures, and acute and chronic A new class of compounds is provided that enables the treatment and / or prevention of neurodegenerative diseases of the present invention, as well as nerve injury caused by ischemia or nerve mediator related diseases or conditions. These disorders are at least in part related to the overactivation of nerve mediator receptors and / or excessive extracellular nerve mediator concentrations.

  First, the present invention relates to a UDP-glucuronosyltransferase for the preparation of a pharmaceutical composition having an inhibitory effect on extracellular nerve mediator release into an individual treated with the pharmaceutical composition. The use of at least one substrate of (UGT) and its salts. According to one particular embodiment, UDP-glucuronosyltransferase (UGT) is expressed in the brain.

  According to a particular embodiment, UDP-glucuronosyltransferase (UGT) is selected in the group consisting of: UDP-glucuronosyltransferase 1A1 (UGT1A1; also referred to as HUG-Br1 or UGT1.1), UDP-glu Chronosyltransferase 1A3 (also referred to as UGT1A3; 1c), UDP-glucuronosyltransferase 1A4 (UGT1A4; also referred to as HUG-Br2), UDP-glucuronosyltransferase 1A6 (UGT1A6), UDP-glucuronosyltransferase 1A9 (UG1; HLUG P4), UDP-glucuronosyltransferase 2B4 (UGT2B4; also called Hlug-25, Th-1, h-1 or h-20) UDP-glucuronosyltransferase 2B7 (UGT2B7), UDP-glucuronosyltransferase 2B10 (UGT2B10; also referred to as h-46), UDP-glucuronosyltransferase 2B15 (UGT2B15; UDPGTh-3, h-3, HLUG-4) (Also referred to as HE 8A), UDP-glucuronosyltransferase 2B17 (UGT2B17), and UDP-glucuronosyltransferase 2B28 (UGT2B28). According to a particular embodiment, UDP-glucuronosyltransferase (UGT) is expressed in the brain and is selected in the group consisting of UDP-glucuronosyltransferase 1A6 (UGT1A6) and UDP-glucuronosyltransferase 2B7 (UGT2B7) .

According to the present invention, the term “UDP-glucuronosyltransferase (UGT)” refers to an enzyme that catalyzes a glucuronidation reaction. This reaction utilizes UDP-glucuronic acid as an auxiliary substrate for the formation of glucuronide. All of these terms are widely used in the literature; for a review, see Turkey and Strassburg, 2000, Annu. Rev. Pharmacol. Toxicol. , 40, 581-616. A preferred method for analyzing the activity of a particular UGT is to express the corresponding cDNA in a cell line that expresses minimal levels of each protein. More specifically, after transfection of the cells by standard transient transfection methods or by stably introducing cDNA into the genome, the cells are selected with an appropriate antibiotic. Then, homogenized the cells in buffer containing Tris -HCl and MgCl _2, sonicated. The cell homogenate is then examined for the ability to glucuronidate a series of substrates. The activity and selectivity of UGT for a substrate can be compared with any known specific substrate (Remmel and Burcell, 1993, Biochem. Pharmacol., 46, 559-566).

Currently, 15 UGT cDNAs, 8 UGT1A proteins encoded by the UGT1A locus, and 7 proteins encoded by the UGT2B gene have been identified in humans. “UDP-glucuronosyltransferase (UGT) 1A6” represents one of the UGT proteins encoded by the UGT1A locus. The human UGT1A6 sequence is shown in SEQ ID NO: 1 (Harding et al., 1988, Proc. Natl. Acad. Sci. 85, 8388-1385). UGT1A6 is also called PNP-UGT, phenol pI6.2, UGT1 ^* 6, UGT1A06 or HLUGP1. “UDP-glucuronosyltransferase (UGT) 2B7” represents one of the UGT proteins encoded by the UGT2B locus. The human UGT2B7 sequence is shown in SEQ ID NO: 2 (Ritter et al., 1990, J. Biol. Chem., 265, 7900-7906. UGT2B7 is Th-2, h-2, Hlug-6, UGT2 ^* 7 or catechol. Also named estrogen UDPGT).

  According to the present invention, the term “UGT substrate” generally refers to an aglycon substrate (ie, a compound that can be enzymatically converted or metabolized by the UGT enzyme disclosed above) and a glucuronidated substrate (ie, via an active group). Both of the compounds converted by the addition of at least one glucuronic acid moiety.

  According to the present invention, the term “neural mediator” is synonymous with “neurotransmitter” and goes beyond the junction (synapse) that separates one nerve cell (neuron) from another nerve cell or muscle. Represents a chemical substance that conveys information. Examples of nerve mediators are acetylcholine (Ach), butyrylcholine (Buch), γ-aminobutyric acid (GABA), serotonin, norepinephrine, epinephrine, endorphin, or catecholamines, especially dopamine, or adenosine triphosphate. According to a preferred embodiment, the nerve mediator is glutamate.

  According to one embodiment, the UGT substrate of the present invention is selected in the group consisting of planar low molecular weight phenols, polycyclic aromatic hydrocarbons, and structurally related compounds. More specifically, the UGT substrate is at least one selected from the group consisting of hydroxyl (alcoholic, phenolic, etc.), carboxyl, sulfuryl, carbonyl and amino (primary, secondary or tertiary) moieties. Contains one part.

  According to a specific embodiment, the substrate is a compound selected from the group consisting of: 1-naphthol, 2-naphthol, 4-nitrophenol, convertible at least by UDP-glucuronosyltransferase 1A6 (UGT1A6) , Methyl salicylate, ketoprofen, naproxen, 5-OH tryptamine / serotonin, carprofen / rimadyl, acetaminophen / paracetamol, benzidine, 4-methylumbelliferone -MU), silymarin (Venketaramanan et al., 2000, Drug Metab Dispos., 28, 1270- ; Silymarin Tokishiforin (toxifolin), silichristin (silichristin), silidianin (silidianin), silybin (silybin) A and B, is a mixture of isosilybin (isosilybin) A and B) [see FIG. 1A and 1B].

  According to another embodiment, the substrate is a compound selected from the group consisting of: transretinoic acid, ASA, AZT, benoxaprofen (convertible), which is convertible by at least UDP-glucuronosyltransferase 2B7 (UGT2B7). benoxaprofen, benzidine, (benzo (a) pyrene mbs), buprenorphine (buprenorphine), carprofen, chloramphenicol, clofibric acid, flavenoids quercetin, chemphenol (kaempfenol), DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone, mefenamic acid (mefenamic acid) ), Phenate NSAID, mycophenolic acid MCPA mofetil, fenoprofen, hydromorphone, ibuprofen, ketoprofen, linoleic acid, lorazepam a lomol e 3 and morphine 6, nalbufen, nalmefene, naltrindole, nalorphine, naloxone, naltrexone, norproxen, norphone Don (oxycodone), oxymorphone, pirlprofen, propanolol, S-oxazepam, tacrolimus, temazepam, ticapronol, c (Valproate), zomepirac, 5-OH tryptamine.

  According to a specific embodiment, the substrate is selected in the group consisting of ketoprofen, naproxen, 5-OH tryptamine / serotonin, carprofen / rimadil, and benzidine.

In certain embodiments, the UGT substrates of the present invention are further substituted with 1 to 4 identical or different heteroatoms and / or heterogroups. Examples of these heteroatoms and / or heterogroups are O, H, alkyl or aryl groups C _n H _{n + 1} (n = 1-5), OCH ₃ , N, halogen (fluorine, chlorine, bromine or iodine atoms ), S, or any labeling element that can visualize the substrate. These substituent atoms or groups and their uses are widely known in the art.

  Addition salts of the substrate of the present invention include inorganic or organic acids or bases such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, perchloric acid, fumaric acid, acetic acid, sodium, lithium, potassium, magnesium, Common salts formed from aluminum, calcium, zinc, ethylenediamine; formic acid, benzoic acid, maleic acid, tartaric acid, citric acid, oxalic acid, aspartic acid, and alkane-sulfonic acids are included.

  UGT substrates used in accordance with the present invention are commercially available in a number of commercial catalogs: for example Merck, “Products chimique et reactifs” 2002, and Acros Organics “Fine Chemicals” 2002-2003:

Other suppliers are:
Naproxen: Apranax (Roche, France) or Naprosyn (Roche, USA);
Carprofen: Limadil (Pfeizer);
Ketoprofen: Toprec (Aventis, France) or Ordis (Wyeth, USA).

  Due to the newly identified inhibitory properties of these UGT substrates, they treat and / or prevent diseases, conditions and seizures, preferably neurological, associated with the deleterious effects of released neurotransmitters. It will be particularly suitable for. In one particular embodiment, the UGT substrate of the present invention is suitable for treating and / or preventing diseases, conditions and seizures, preferably neurological, associated with the detrimental effects of released glutamate.

  According to one particular embodiment, the diseases, conditions and seizures are associated with the deleterious effects of excessively released neurotransmitters, more specifically the deleterious effects of excessively released glutamate.

  Accordingly, the present invention further provides a method for treating and / or preventing cytotoxicity induced by neuronal mediators in a patient in need thereof, comprising at least one of the above UDP-glucuronosyltransferases (UGT). A method comprising administering to a patient a composition comprising a substrate (therapeutically effective amount) and a pharmaceutically acceptable carrier. In certain embodiments, the UGT substrate is a compound convertible by at least one UDP-glucuronosyltransferase (UGT), preferably at least one UDP-glucuronosyltransferase (UGT) expressed in the brain.

  Preferably, the invention is a method for treating and / or preventing glutamate-induced cytotoxicity in a patient in need thereof, comprising at least one substrate of UDP-glucuronosyltransferase (UGT) as described above It relates to a method comprising administering to a patient a composition comprising a (therapeutically effective amount) and a pharmaceutically acceptable carrier. In a particular embodiment, the UGT substrate is a compound convertible by at least one UDP-glucuronosyltransferase (UGT), preferably at least one UDP-glucuronosyltransferase (UGT) expressed in the brain.

  According to another aspect, the UGT substrate is selected in the group consisting of planar low molecular weight phenols, polycyclic aromatic hydrocarbons, and structurally related compounds. More specifically, the UGT substrate is at least one selected from the group consisting of hydroxyl (alcoholic, phenolic, etc.), carboxyl, sulfuryl, carbonyl and amino (primary, secondary or tertiary) moieties. Contains one part.

  According to another embodiment, the substrate is a compound selected from the group consisting of: 1-naphthol, 2-naphthol, 4-nitrophenol, convertible by UDP-glucuronosyltransferase 1A6 (UGT1A6) Methyl salicylate, ketoprofen, naproxen, 5-OH tryptamine / serotonin, carprofen / rimadyl, acetaminophen / paracetamol, benzidine, 4-methylumbelliferone (4-MU), silymarin (Venketaramanan et al., 2000, Drug Metab Dispos , 28, 1270-3; silymarin is a mixture of toxifolin, silicristine, silydianin, silybin A and B, and isosiribine A and B) [see FIG.

  According to another embodiment, the substrate is a compound selected from the group consisting of: transretinoic acid, ASA, AZT, beoxaprofen, benzidine, convertible by UDP-glucuronosyltransferase 2B7 (UGT2B7). (Benzo (a) pyrene mbs), buprenorphine, carprofen, chloramphenicol, clofibric acid, flavenoids quercetin, chemphenol, cyclosporine, DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone, mefenamic acid, phenemate NSAID , Mycophenolic acid MCPA mofetil, fenoprofen, hydromorphone, ibuprofen, ketoprofen, linoleic acid, lorazepam, losartan, menthol, morphine 3 and mol -6, nalbufen, nalmefene, naltoldol, nalolphine, naloxone, naltrexone, S-naproxen, norcodeine, normorphone, oxycodone, oxymorphone, pyrprofen, propanolol, S-oxazepam, tacrolimus, temazepam, tolcapone, thiaprofenic, valproate, valproate Zomepirac, 5-OH tryptamine.

  According to a particular embodiment, the substrate is selected in the group consisting of ketoprofen, naproxen, 5-OH tryptamine / serotonin, carprofen / rimadil, and benzidine.

  In the context of the present invention, the term “neurotransmitter-induced cytotoxicity” or “neurotransmitter-induced cytotoxicity” refers to cytotoxicity associated with overactivation of the involved neuromediator receptor. To do. These terms are well known to those skilled in the art. More specifically, “glutamate-induced cytotoxicity” refers to all diseased cells that express the glutamate receptor. According to a preferred embodiment, these cells that are susceptible to nerve mediators are nervous cells (preferably neuro-cells), preferably neurons. These affected nervous system cells are present, for example, in the brain, spinal cord, retina, nerve-muscle junction, and the like. “Cytotoxicity” means that cell function and / or properties are impaired, resulting in cell dysfunction and ultimately cell death.

  In a particularly preferred embodiment, the method of the invention is for the treatment and / or prevention of neurotoxicity induced by a nerve mediator, more preferably for the treatment and / or prevention of neurodegeneration (i.e. degeneration of neural cells). is there.

  In another particularly preferred embodiment, the method of the invention is for the treatment and / or prevention of glutamate-induced neurotoxicity, more preferably for the treatment and / or prevention of neurodegeneration (ie, degeneration of neural cells). is there.

  The present invention further provides a method for modulating the release of at least one nerve mediator in a patient comprising at least one substrate (therapeutically effective amount) of UDP-glucuronosyltransferase (UGT) and pharmaceutically. Relates to a method comprising administering to a patient a composition containing an acceptable carrier, wherein the substrate is as detailed above. In a particular embodiment, the UGT substrate is a compound convertible by at least one UDP-glucuronosyltransferase (UGT), preferably at least one UDP-glucuronosyltransferase (UGT) expressed in the brain.

  The present invention further comprises a method for modulating glutamate release in a patient, comprising at least one substrate (therapeutically effective amount) of UDP-glucuronosyltransferase (UGT) and a pharmaceutically acceptable carrier. Relates to a method comprising administering the composition to a patient; wherein the substrate is as detailed above. In a particular embodiment, the UGT substrate is a compound convertible by at least one UDP-glucuronosyltransferase (UGT), preferably at least one UDP-glucuronosyltransferase (UGT) expressed in the brain.

  “Modulating nerve mediator release” means that the level of nerve mediator release in an untreated patient differs from that seen after treatment of the patient with a substrate of the present invention. According to one embodiment, treatment of a patient with a substrate of the invention results in a negative modulation, preferably inhibition of release by nerve mediator producing cells, so that the concentration of nerve mediator in the treated patient is seen prior to the treatment. Compared to the concentration of the same nerve mediators.

  The present invention further provides a method for treating and / or preventing a disease and / or condition associated with excessive release of at least one nerve mediator in a patient comprising at least a UDP-glucuronosyltransferase (UGT). Relates to a method comprising administering to a patient a composition comprising one substrate (therapeutically effective amount) and a pharmaceutically acceptable carrier; wherein the substrate is as detailed above. In a particular embodiment, the UGT substrate is a compound convertible by at least one UDP-glucuronosyltransferase (UGT), preferably at least one UDP-glucuronosyltransferase (UGT) expressed in the brain.

  The present invention further provides a method for treating and / or preventing diseases and / or conditions associated with excessive release of glutamate in a patient comprising at least one substrate (therapy) of UDP-glucuronosyltransferase (UGT). Effective amount) and a composition comprising administering to the patient a composition containing a pharmaceutically acceptable carrier, wherein the substrate is as detailed above. In a particular embodiment, the UGT substrate is a compound convertible by at least one UDP-glucuronosyltransferase (UGT), preferably at least one UDP-glucuronosyltransferase (UGT) expressed in the brain.

  According to the present invention, the term “treatment and / or prevention” refers to an act intended to cause a beneficial change in the condition of a mammal, eg a human (sometimes referred to as a patient). Beneficial changes can include, for example, one or more of the following: restoration of function, reduction of symptoms, restriction or delay of progression of the disease, disorder or condition, or prevention of worsening of the patient's condition, disease or disorder. Limit or delay. Such therapies can include, for example, nutritional modifications, radiation administration, drug administration, habit modification, and combinations thereof, for example.

  According to the present invention, “disease and / or condition associated with excessive release of at least one nerve mediator” refers to a number of acute and chronic nerve mediator related diseases or conditions, in particular neurological diseases. To do. More specifically, it refers to epileptic seizures and acute and chronic neurodegenerative diseases, as well as nerve injury caused by ischemia or nerve mediator related diseases or conditions, wherein the disorders are at least partially Is associated with overactivation of neuronal mediator receptors and / or excessive extracellular nerve mediator concentrations. Examples include those with chronic or acute degenerative diseases such as Alzheimer's disease, Huntington's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA). ), Retinal disorders, stroke, clinical depression, and traumatic brain disorders, which involve neuronal cell death caused by overstimulation of nerve mediator receptors, particularly glutamate receptors. Similarly, neurological injury caused by ischemia or drug-induced neurotoxicity, such as the neurotoxic effect of methamphetamine (METH) on striatal dopaminergic neurons is also an indication of the method according to the invention. Other indications include, for example, pain, hormone balance, blood pressure, thermoregulation, breathing, learning, pattern recognition or memory-related conditions, or disorders associated with hypoxia or hypoglycemia, especially those indications that are glutamate This is the case of the related state.

  Examples include treatment of adverse effects due to excessive glutamate release resulting from epilepsy, clinical depression, amyotrophic lateral sclerosis, spinal muscular atrophy (SMA), Huntington's disease, trauma and other angiogenic brain accidents. It is done.

  In the embodiments of the invention disclosed above, most compositions and methods relate to the substrate in aglycone form. Alternatively, the present invention further provides at least one UDP-glucuronosyltransferase (UGT) for preparing a pharmaceutical composition having an inhibitory effect on extracellular nerve mediator release into an individual treated with the pharmaceutical composition. Of glucuronic acid conjugated substrates and their salts. According to this aspect, it is understood that the substrate for UDP-glucuronosyltransferase (UGT) is glucuronidated prior to administration to a patient in need thereof. The glucuronidation can be achieved by modifying the aglycon substrate by chemical or enzymatic methods. Chemical glucuronidation can be accomplished using standard chemical methods well known in the art, including, for example, adding a glucuronide group to the aglycon substrate to be used (Lacy and Sainsbury, 1995, Tetrahedron Lett., 36, 3949-50). Alternatively, the glucuronidated substrate can be obtained by glucuronidation with an enzyme using at least one UDP-glucuronosyltransferase (UGT) capable of glucuronidating an aglycon substrate. In a preferred embodiment, the UDP-glucuronosyltransferase (UGT) is naturally expressed in the brain, such as UDP-glucuronosyltransferase (UGT) 1A6 or UDP-glucuronosyltransferase (UGT) 2B7.

  According to one embodiment, the glucuronidated UGT substrate comprises a planar small molecule glucuronidated phenol, a glucuronidated polycyclic aromatic hydrocarbon, and a structurally related compound. Selected in group. More specifically, the glucuronidated UGT substrate is a hydroxyl (alcoholic, phenolic, etc.), carboxyl, sulfuryl, carbonyl and amino (primary, secondary or tertiary) attached to the glucuronic acid moiety. ) At least one portion selected in the group consisting of portions.

  According to a specific embodiment, the glucuronidated substrate is 1-naphthol, 2-naphthol, 4-nitrophenol, methyl salicylate, ketoprofen, naproxen, 5-OH tryptamine / serotonin, carprofen / limadil, acetaminophen / Paracetamol, benzidine, 4-methylumbelliferone (4-MU), silymarin (Venketaramanan et al., 2000, Drug Metab Dispos., 28, 1270-3; silymarin is toxifolin, silicristine, silydinine, silybinin A and B, Selected from the group consisting of isociribine A and B) and further glucuronidated.

  According to another embodiment, the glucuronidated substrate is trans retinoic acid, ASA, AZT, benoxaprofen, benzidine, (benzo (a) pyrene mbs), buprenorphine, carprofen, chloramphenicol, clofibulin. Acids, flavenoids quercetin, chemphenol, cyclosporine, DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone, mefenamic acid, phenate NSAID, mycophenolate MCPA mofetil, fenoprofen, hydromorphone, ibuprofen, ketoprofen, linoleic acid, lorazepam , Losartan, menthol, morphine 3 and morphine 6, nalbufen, nalmefene, naltrindole, nalolphine, naloxone, naltrexone, S- Selected in the group consisting of proxene, norcodeine, normorphone, oxycodone, oxymorphone, pyrprofen, propanolol, S-oxazepam, tacrolimus, temazepam, tolcapone, thiaprofenic, valproate, zomepirac, 5-OH tryptamine, and further conjugated with glucuronic acid Yes.

  Glucuronic acid-conjugated substrate addition salts of the present invention include inorganic or organic acids or bases such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, perchloric acid, fumaric acid, acetic acid, sodium, lithium , Potassium, magnesium, aluminum, calcium, zinc, ethylenediamine; common salts formed from formic acid, benzoic acid, maleic acid, tartaric acid, citric acid, oxalic acid, aspartic acid, and alkane-sulfonic acids.

  The present invention further provides a method for treating and / or preventing neuronal mediator-induced cytotoxicity in a patient in need thereof, comprising at least one UDP-glucuronosyltransferase (UGT) as described above. It relates to a method comprising administering to a patient a composition comprising a glucuronidated substrate (therapeutically effective amount) and a pharmaceutically acceptable carrier.

  The present invention further provides a method for treating and / or preventing glutamate-induced cytotoxicity in a patient in need thereof, comprising at least one glucuronic acid of the above UDP-glucuronosyltransferase (UGT). It relates to a method comprising administering to a patient a composition comprising a conjugated substrate (therapeutically effective amount) and a pharmaceutically acceptable carrier.

  In particularly preferred embodiments, the “nerve-mediated cytotoxicity” is a neuro-mediated neurotoxicity, more preferably neurodegeneration (ie, degeneration of nervous system cells).

  In another particularly preferred embodiment, “glutamate-induced cytotoxicity” is glutamate-induced neurotoxicity, more preferably neurodegeneration (ie, degeneration of nervous system cells).

  The present invention further provides a method for modulating the release of at least one nerve mediator in a patient comprising at least one glucuronidated substrate of UDP-glucuronosyltransferase (UGT) (therapeutically effective amount). ) And a composition containing a pharmaceutically acceptable carrier, wherein the substrate is as detailed above.

  The present invention is further a method for modulating glutamate release in a patient comprising at least one glucuronidated substrate (therapeutically effective amount) of UDP-glucuronosyltransferase (UGT) and a pharmaceutically acceptable A method comprising administering to a patient a composition containing a possible carrier; wherein the substrate is as detailed above.

  The present invention further provides a method for treating and / or preventing a disease and / or condition associated with excessive release of at least one nerve mediator in a patient comprising at least a UDP-glucuronosyltransferase (UGT). Relates to a method comprising administering to a patient a composition comprising one glucuronidated substrate (therapeutically effective amount) and a pharmaceutically acceptable carrier, wherein the substrate is as detailed above .

  The present invention is further a method for treating and / or preventing diseases and / or conditions associated with excessive release of glutamate in a patient, comprising at least one glucuronidation of UDP-glucuronosyltransferase (UGT). A method comprising administering to a patient a composition comprising a formulated substrate (therapeutically effective amount) and a pharmaceutically acceptable carrier, wherein the substrate is as detailed above.

  The UGT substrates described in the present invention (including aglycone or glucuronidated forms) are administered as a composition containing at least one active compound and a pharmaceutically acceptable carrier. Any common pharmaceutically acceptable carrier can be used in preparing such compositions. The carrier material may be an organic or inorganic inert carrier material suitable for the chosen route of administration. Suitable carriers include water, gelatin, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, petrolatum and the like. Furthermore, the pharmaceutical composition may contain other medicinal substances. Additional additives such as flavoring agents, preservatives, stabilizers, emulsifiers, salts that alter osmotic pressure, buffers and the like may be added according to accepted pharmaceutical formulation practices. Any common dosage form can be used, such as tablets, capsules, pills, powders, granules. Advantageously, they are in the form of tablets, dragees, hard gelatin capsules, capsules, granules, or solutions or suspensions for administration by the injectable route for oral administration.

  The methods of the invention can be practiced by administering a composition containing a substrate of the invention (including aglycone or glucuronidated forms) by any route by which the drug is generally administered. Such routes include systemic routes and local routes. Examples are intravenous, intramuscular, subcutaneous, intracranial, intraventricular, inhalation (Gradinaru et al., 1999, supra), intraperitoneal, and oral routes. Preferably, the method of the present invention is performed by an oral or intravenous route of administration.

  In accordance with the present invention, the substrates described herein (including aglycone or glucuronidated forms) are useful in pharmaceutically acceptable oral regimes. Preferred oral dosage forms include hard or soft gelatin, methylcellulose or other suitable material tablets, capsules that dissolve easily in the gastrointestinal tract. Oral dosage forms contemplated according to the present invention will vary depending on the requirements of the individual patient and will be determined by the prescribing physician. A preferred oral dosage form is a capsule or tablet containing 50-500 mg of the substrate of the present invention.

  A typical formulation for intravenous administration is a sterile aqueous solution containing a water / buffer solution. Intravenous vehicles include supplemental fluids, nutrients, and electrolytes. Preservatives and other additives may be present, for example, as antibiotics and antioxidants. Compositions for intravenous bolus administration can contain up to 10 mg / ml (10,000 mg / liter) of the substrate described herein. Compositions for intravenous administration preferably contain from about 50 mg / liter to about 500 mg / liter of at least one substrate described herein.

  In practicing the methods of the invention, the substrate of the invention (including aglycone or glucuronidated forms) is generally administered to an adult daily in an amount of about 5 mg / kg to about 30 mg / kg daily in a single dose or split. The amount is preferably about 13 mg / kg to about 17 mg / kg per day, preferably administered orally or intravenously. However, the exact dose will vary depending on patient requirements. The dose is applied according to the patient to be treated and the pathological condition, for example 1-100 mg / day. This therapy is generally performed for about 3 months. Alternatively, the method of the invention can be carried out prophylactically for an indefinite period of time in patients at high risk for acute neurotoxic events such as stroke. For the treatment of acute neurotoxic events, according to the method of the invention as soon as possible after diagnosis of acute neurotoxic events, preferably within 12 hours, most preferably within 6 hours after the onset of acute neurotoxic events. The patient should be treated. When drugs are administered orally, they are generally administered at regular intervals.

These drugs are administered in particular by the oral or injection route.
According to one particular embodiment, the UGT substrates described in the present invention (including aglycone or glucuronidated forms) are modified with specific groups that promote the passage of the UGT substrate through the blood brain barrier (BBB). One mode is lipidization, which involves adding a lipid-like group by modification of the hydrophilic portion of the substrate structure. The resulting lipid soluble substrate is transported through the BBB by utilizing the pores that are transiently formed in the lipid bilayer. Another approach is the addition of hydrophobic groups to enhance BBB transport by passive diffusion. For example, addition of methyl groups or acetylation of amine groups increases lipophilicity and brain permeability. Yet another method is to modify the substrate so that the compound has a structure resembling a nutrient and utilize one of the specific carrier-mediated transport systems within the BBB, such as amino acids, hexoses, vitamins and neuropeptides. It is. Another method is to attach a substrate via a chemical linker to a small synthetic peptide vector that efficiently crosses the cell membrane, such as Pegelin or Penetratin (for a general review, see Temsanani). et al., 2000, Pharm. Sci. Technol. Today, 3, 155-162).

  According to another aspect, a patient in need of a substrate or formulation of the invention (including aglycone or glucuronidated forms) in combination with a second compound or formulation intended to facilitate transport through the BBB To be administered. For example, a substrate or formulation of the invention is combined with hypertonic solutions, bioactive agents such as bradykinin and angiotensin peptides, vasoactive agents such as histamine, leukotrienes with the ability to transiently disrupt BBB. According to the present invention, “in combination” or “in combination with” refers to the substrate or formulation of the present invention and a second compound or formulation intended to facilitate transport through the BBB, simultaneously or sequentially, or It means that it can be used at different times. Simultaneous means that they are administered together. In this case, these two essential ingredients can be mixed prior to administration to prepare a composition, or can be administered simultaneously to a patient in need thereof. They can also be administered sequentially, ie one after the other regardless of which component is administered first. In addition, timed or intermittent modes of administration can be employed, which can be stopped and restarted at regular or irregular intervals. It is pointed out that the route of administration and site of administration of the two components may be different. The interval between administration times and the route and site of administration can be determined by those skilled in the art. An interval of 10 minutes to 72 hours, advantageously 30 minutes to 48 hours, preferably 1 to 24 hours, very preferably 1 to 6 hours can be recommended.

  Pharmacogenomic analysis (see, eg, Ciotti et al., 1997, Pharmacogenetics, 7, 485-495) impairs enzyme activity due to polymorphism of the UGT gene and reduces or increases patient responsiveness to UGT substrates. It was shown that it was possible. For example, missense mutations in UGT1A6 (A541 → G541, and A552 → C552) that mutate UGT1A6 activity and thus their substrate metabolic rate have been identified (for more details see Chiotti et al., 1997, Pharmacogenetics, 7, 485-495). Thus, according to another particular aspect, the therapeutic and / or prophylactic methods of the invention further comprise a preliminary comprising determining whether the patient being treated exhibits a genetic polymorphism that results in a decrease or enhancement of UGT activity. Process. More specifically, this preliminary step is performed by nucleotide sequence analysis of the genomic DNA of the patient, wherein the UGT1A6 gene of the patient to be treated is at least one of mutation A541 → G541 (T181A mutation) or mutation A552 → C552 (R184S mutation). Is to determine whether one is included. Details of this method are described in, for example, Chiotti et al. 1997, Pharmacogenetics, 7, 485-495, the entire contents of which are incorporated herein by reference.

  All publications and patent applications cited herein are hereby incorporated by reference as if each publication and patent application was specifically and individually indicated to be incorporated. Although the invention has been described in detail by way of illustration and specific examples so as to provide a clear understanding of the invention, in light of the teachings of the invention, certain changes and modifications may be made thereto without departing from the spirit and scope of the claims. It will be obvious to those skilled in the art.

  Although the present invention has been specifically described, it is to be understood that the terminology used is for purposes of illustration and not limitation. Obviously, many modifications and variations of the present invention are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims, the invention may be practiced otherwise than as specifically described. Accordingly, those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments specifically described herein. Such equivalents are intended to be encompassed in the scope of the claims.

These and other aspects are disclosed in, or are apparent from, the first description and examples herein. Other references relating to any of the methods, uses and compounds used in accordance with the present invention can be retrieved from public libraries using, for example, electronic devices. For example, the public database “Medline” that can be used on the Internet under http://www.ncbi.nlm.nih.gov/PubMed/medline.html can be used. Other databases and addresses such as http://www.ncbi.nlm.nih.gov/, http://www.infobiogen.fr/, http://www.fmi.ch/biology/research_tools.html, http : //www.tigr.org/ is known to those skilled in the art and can also be obtained, for example, using http://www.lycos.com. A review of biotechnology patent information and related patent information sources useful for retrospective and current recognition is given in Berks, TIBTECH 12 (1994), 352-364.
[Example]
The cortex was detached from cortical neuron primary culture Wistar rat fetus (E16) and kept in PBS without calcium and magnesium. They were then treated for 15 minutes at 37 ° C. in PBS containing 0.25% trypsin and placed in complete medium containing horse serum for trypsin activity inhibition. The cortex was then dissociated in complete medium (Neurobasal, 2% horse serum, 2 mM glutamine, supplemental B27 (1 ×), 100 μg / ml gentamicin, 10 μM β-mercaptoethanol). Cells were seeded at 10 ⁵ cells / well in a 96-well plate pre-coated with 9 μg / ml of polyornithine (Poly-Ornitine). The culture was kept at 37 ° C. in a humid incubator with 5% CO ₂ . On the second day, 5 μM cytosine arabinofuranoside was added to the cell culture to prevent glial cell growth. On day 7, primary cultures of neurons used for subsequent experiments were obtained.

Evaluation of neuronal cell death Neuronal cell death was estimated by measuring the release of cytosolic enzyme, lactate dehydrogenase (LDH), into the culture medium of the cell culture. Quantification of LDH release was performed using a colorimetric CytoTox96® non-radioactive assay (Promega). In summary, 100 μL of cell culture medium was removed and centrifuged at 1500 rpm for 4 minutes to remove cell debris. 50 μL was then added to 50 μL assay buffer and placed in the dark for 30 minutes at room temperature. The reaction was stopped with 50 μL of the stop solution, and the absorbance at 490 nm was measured. The percent cell death was calculated by comparison with the control.

Evaluation of glutamate release Glutamate release by cultured neurons was evaluated in chemiluminescent enzyme assays in aliquot supernatants (Israel et al., Neurochem Int. January 1993; 22 (1): 53-8).

Neuroprotection assay Diazepam (Sigma) and a glucuronide derivative of diazepam, called temazepam (Alltech), and a derivative that is a UDP-glucuronosyltransferase 2B7 (UGT2B7) substrate, induces UGT2B7 substrate by glutamate Prophylactic and / or therapeutic effects on neurological disorders that have been demonstrated have been demonstrated.

Diazepam and temazepam were used to protect primary cultures of neurons isolated from rat cerebral cortex from neurotoxin-induced cell death.
400 μM 3-nitropropionic acid (3-NP, Sigma), a toxin that induces epileptic seizures in animal models, in vitro in the absence or presence of 0.1, 1 and 5 μM diazepam and temazepam Was processed. After 24 hours, the level of cell death of the cell culture induced by toxin was analyzed by lactate dehydrogenase release quantification.

  The results (FIG. 2) show that diazepam protects neurons from 3-NP at all test concentrations, whereas temazepam significantly enhances this cytoprotection at 1 and 5 μM by about 50%. These data show that glucuronide modification of diazepam affects cell viability when exposed to toxic compounds.

  Similar results were obtained with other neurotoxins, ie, 100 μM kainic acid (Fisher Bioblock) that mimics glutamate-induced epileptic seizures in animal models. For 1 and 5 μM temazepam, better neuroprotection was seen than the same concentration of diazepam (FIG. 3).

Neuroprotection Assay 1-Naphthol (Merck), a substrate of UDP-glucuronosyltransferase 1A6 (UGT1A6), was used to demonstrate the effect of UGT1A6 substrate on glutamate-induced neurological damage.

1-Naphthol was used to protect primary cultures of neurons isolated from rat cerebral cortex from neurotoxin-induced cell death.
5 μM ionomycin (Ionmycin, Sigma), ie, calcium ionophore, in the presence of 10 μM 1-naphthol, 5 or 10 μM 3-methylcholanthrene (3-methylcholanthrene, 3-MC, Aidrich), ie 1-naphthol. Neurons were treated in vitro in the absence or presence of UGT inducers that allow in situ glucuronidation.

  The results (FIG. 4) show that naphthol protects neurons from ionomycin-induced cell death and that glucuronidation of 1-naphthol by 3-MC enhances the protective effect of 1-naphthol in a dose-dependent manner Indicates to do.

These data indicate that the glucuronide derivative of the UGT1A6 substrate provides a better neuroprotective effect than the parent molecule.
Measurement of glutamate concentration Glutamate released by neurons was followed by an enzyme assay coupled to a chemiluminescent reaction. In this experiment, primary cultures of neurons isolated from rat cerebral cortex were treated with 10 μM 1-naphthol in the presence or absence of 1 or 5 μM 3-MC. The results (FIG. 5) show that 1-naphthol reduces glutamate release and that glucuronidation of 1-naphthol by 3-MC enhances glutamate reduction in a dose-dependent manner.

  This experiment shows that the reduction in glutamate release is improved with the protective effect of glucuronidation on the UGT1A6 substrate.

The chemical structures of the compounds cited and shown throughout this specification are shown. The chemical structures of the compounds cited and shown throughout this specification are shown. 2 shows the effect of diazepam and temazepam on neuronal cell death. Neuronal cell death in culture was arbitrarily set to zero. Cells were treated with vehicle (row 1) and 400 μM 3-NP alone (row 2), or at the same time, diazepam (rows 3-5) and temazepam (rows 6-8) at any one of three test concentrations. Processed. The results show that except for 0.1 μM (row 6), 1 μM (row 7) and 5 μM (row 8) temazepam improve the neuroprotective effect seen at all concentrations of diazepam (rows 3-5). Indicates. 2 shows the effect of diazepam and temazepam on neuronal cell death. Neuronal cell death in culture was arbitrarily set to zero. Cells were treated with vehicle (row 1) and 100 μM kainic acid alone (row 2) or simultaneously with any one of three test concentrations of diazepam (rows 3-5) and temazepam (rows 6-8). did. The results show that except for 0.1 μM (row 6), 1 μM (row 7) and 5 μM (row 8) temazepam improve the neuroprotective effect seen at all concentrations of diazepam (rows 3-5). Indicates. 1 shows the effect of 1-naphthol and 1-naphthol glucuronide on neuronal cell death. Neuronal cell death in culture was arbitrarily set to zero. Cells were treated with vehicle (row 1) and 5 μM ionomycin alone (row 4) or simultaneously with 10 μM 1-naphthol (row 7) and 3-methylcholanthrene 5 μM (row 8) or 10 μM (row 9). did. These two test doses of 3-methylcholanthrene were evaluated in the absence (rows 2, 3) or presence (rows 5, 6) of ionomycin and considered as controls. The results show that 3-methylcholanthrene alone does not affect the natural death of neurons (columns 2, 3) and has no protective effect against ionomycin-induced toxicity (columns 5, 6). . The results also show that the protection afforded by 10 μM 1-naphthol (row 7) is improved by glucuronidation induced by 3-methylcholanthrene 5 μM (row 8) and 10 μM (row 9). 1 shows the effect of 1-naphthol and 1-naphthol glucuronide on glutamate released by neurons in culture. Glutamate release was assessed by a chemiluminescent enzyme assay. Basal levels of glutamate release were quantified in untreated cells (Row 1), cells treated with vehicle (Row 2), and cells treated with 3-methylcholanthrene at the highest concentration tested in this experiment (Row 3). did. The effects of 1-naphthol 1 μM (row 4) and 10 μM (row 7) were evaluated in the presence of 3-methylcholanthrene 1 μM (rows 5, 8) and 5 μM (rows 6, 9). The results show that the vehicle and glucuronidation inducer 3-methylcholanthrene do not affect glutamate release (columns 2 and 3), whereas 1-naphthol reduces glutamate release at both concentrations (columns 4 and 7). ) Furthermore, for the two test doses of 3-methylcholanthrene, glutamate conjugation is improved by conjugation of both doses of 1-naphthol (rows 5, 6, 8, 9). This improvement is statistically significant at the highest concentration of 3-methylcholanthrene (rows 6, 9) at any 1-naphthol dose.

[Sequence Listing]

Claims (15)

  Use of at least one substrate of UDP-glucuronosyltransferase (UGT) and salts thereof for the preparation of a pharmaceutical composition having an inhibitory effect on extracellular glutamate release into an individual treated with the pharmaceutical composition.   The UDP-glucuronosyltransferase (UGT) is expressed in the brain and is selected in the group consisting of UDP-glucuronosyltransferase 1A6 (UGT1A6) and UDP-glucuronosyltransferase 2B7 (UGT2B7). use.   Use according to claim 1 or 2, wherein the UGT substrate is selected in the group consisting of planar low molecular weight phenols, polycyclic aromatic hydrocarbons, and structurally related compounds.   The substrate is a substrate of UDP-glucuronosyltransferase 1A6 (UGT1A6), 1-naphthol, 2-naphthol, 4-nitrophenol, methyl salicylate, ketoprofen, naproxen, 5-OH tryptamine / serotonin, carprofen / limadil, acetamino 4. Use according to any one of claims 1 to 3, selected in the group consisting of phen / paracetamol, benzidine, 4-methylumbelliferone (4-MU), silymarin.   The substrate is a substrate of UDP-glucuronosyltransferase 2B7 (UGT2B7), transretinoic acid, ASA, AZT, beoxaprofen, benzidine, (benzo (a) pyrene mbs), buprenorphine, carprofen, chloramphenicol, Clofibric acid, flavenoids quercetin, chemphenol, cyclosporine, DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone, mefenamic acid, phenemate NSAID, mycophenolic acid MCPA mofetil, fenoprofen, hydromorphone, ibuprofen, ketoprofen, linoleic acid , Lorazepam, losartan, menthol, morphine 3 and morphine 6, nalbufen, nalmefene, naltrindole, nalolphine Selected from the group consisting of naloxone, naltrexone, S-naproxen, norcodeine, normorphone, oxycodone, oxymorphone, pyrprofen, propanolol, S-oxazepam, tacrolimus, temazepam, tolcapone, thiaprofenic, valproate, zomepirac, 5-OH tryptamine The use according to any one of claims 1 to 3.   Use according to any one of claims 1 to 5, wherein the pharmaceutical composition is for treating and / or preventing glutamate-induced cytotoxicity in a patient in need thereof.   The use according to any one of claims 1 to 5, wherein the pharmaceutical composition is for treating and / or preventing glutamate-induced neurotoxicity.   The use according to any one of claims 1 to 5, wherein the pharmaceutical composition is for treating and / or preventing neurodegeneration.   6. Use according to any one of claims 1 to 5, wherein the pharmaceutical composition is for modulating glutamate release in a patient.   The use according to any one of claims 1 to 5, wherein the pharmaceutical composition is for treating and / or preventing diseases and / or conditions associated with excessive release of glutamate in a patient.   Diseases and / or conditions associated with excessive release of glutamate include epileptic seizures, acute and chronic neurodegenerative diseases, ischemia, Alzheimer's disease, Huntington's disease, Parkinson's disease, multiple sclerosis (MS), amyotrophic side Cord sclerosis (ALS), spinal muscular atrophy (SMA), retinal disorders, stroke and traumatic brain disorders, drug-induced neurotoxicity, pain, hormone balance, blood pressure, thermoregulation, breathing, learning, pattern recognition, memory, And use according to claim 10 selected in the group consisting of disorders associated with hypoxia or hypoglycemia.   At least one glucuronidated substrate of UDP-glucuronosyltransferase (UGT) for the preparation of a pharmaceutical composition having an inhibitory effect on extracellular glutamate release into an individual treated with the pharmaceutical composition; Use of its salts.   The glucuronidated UGT substrate is selected in the group consisting of planar small molecule glucuronidated phenols, glucuronidated polycyclic aromatic hydrocarbons, and structurally related compounds; Use according to claim 12.   Glucuronidated substrates are 1-naphthol, 2-naphthol, 4-nitrophenol, methyl salicylate, ketoprofen, naproxen, 5-OH tryptamine / serotonin, carprofen / limadil, acetaminophen / paracetamol, benzidine, 4-methyl The use according to any one of claims 12 to 13, which is selected in the group consisting of umbelliferone (4-MU) and silymarin and is further glucuronidated.   Glucuronic acid-conjugated substrates are trans retinoic acid, ASA, AZT, benoxaprofen, benzidine, (benzo (a) pyrene mbs), buprenorphine, carprofen, chloramphenicol, clofibric acid, flavenoids quercetin, chem Phenol, cyclosporine, DMXAA, diclofenac, dihydrocodeine DHC, dihydromorphone, mefenamic acid, phenemate NSAID, mycophenolic acid MCPA mofetil, fenoprofen, hydromorphone, ibuprofen, ketoprofen, linoleic acid, lorazepam, losartan, menthol, morphine 3 And morphine 6, nalbufen, nalmefene, naltrindole, nalolphine, naloxone, naltrexone, S-naproxen, nor Selected from the group consisting of dein, normorphone, oxycodone, oxymorphone, pyrprofen, propanolol, S-oxazepam, tacrolimus, temazepam, tolcapone, thiaprofenic, valproate, zomepirac, 5-OH tryptamine, and further conjugated with glucuronic acid, Use according to any one of claims 12-13.