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    • 专利摘要:
    Triazoles for the regulation of intracellular calcium homeostasis. The present invention relates to 1,2,3-triazoles of the formula: {image-01} Useful to improve or restore the function of intracellular calcium homeostasis and the ryr-calstabin binding in human and animal cells. It is also related to methods of synthesizing said compounds, to pharmaceutical compositions containing them, and to their use for preventing or treating musculoskeletal, cardiac and nervous system disorders. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2643856A1
    申请号:ES201630670
    申请日:2016-05-24
    公开日:2017-11-24
    发明作者:Jesús María Aizpurua Iparraguirre；Aitziber IRASTORZA EPELDE；Pablo FERRÓN CELMA；José Ignacio Miranda Murua；Ainara Vallejo Illarramendi；Adolfo José LÓPEZ DE MUNAIN ARREGUI；Iván TORAL OJEDA；Garazi ALDANONDO ARISTIZABAL
    申请人:Euskal Herriko Unibertsitatea；Administracion General de la Comunidad Autonoma de Euskadi；
    IPC主号:
    专利说明:

    Triazoles for the regulation of intracellular calcium homeostasis Field of the Invention
    The present invention relates to new substituted 1,2,3-triazoles useful for improving
    5 or restore the function of intracellular calcium homeostasis in human and animal cells. It is also related to methods of synthesis of said compounds, pharmaceutical compositions containing them, and their use to prevent or treat musculoskeletal, cardiac and neurodegenerative disorders. 10 Background of the invention
    Muscular dystrophies are hereditary heterogeneous diseases that are characterized by weakness and progressive atrophy of skeletal muscle. Duchenne muscular dystrophy (DMD) is one of the most frequent forms, it is linked to the X chromosome and occurs in 1 in 3500 males. As in the case of its most benign allelic form (Becker muscular dystrophy,
    15 BMD), both are produced by mutations in the gene encoding dystrophin, a 427-kDa cytoskeleton protein. Genetic studies are not enough for the eradication of the disease due to the high incidence of sporadic cases, so the search for new effective therapies is urgently needed.
    Waist muscular dystrophies (LGMDs) are a large group of muscular dystrophies
    20 inherited characterized by progressive proximal weakness with predominant involvement of the pelvic and shoulder girdles. Within the recessive forms of LGMD, calpainopathy or LGMD2A is the most frequent form and is caused by mutations in the gene that encodes calpain 3 (CAPN3), a non-lysosomal cysteine protease necessary for the proper functioning and regeneration of muscle.
    25 Myotonic dystrophy type 1 (DM1) is the most common adult form of muscular dystrophy and is characterized by muscular weakness, myotonia and multisystemic involvement. It is an autosomal dominant inherited disease caused by an unstable expansion of the CTG triplet repeat located in the 3 'non-coding region of the DMPK gene, located on the long arm of chromosome 19 and expressed predominantly in skeletal muscle.


    The above mentioned as well as other muscular dystrophies (such as DM2, recessive and dominant LGMD, congenital or metabolic myopathies among others) share alterations in intracellular calcium homeostasis. Because the alteration of the intracellular concentration of Ca2 + in muscle fibers seems to represent a common central pathogenic mechanism, the development of therapeutic interventions that prevent alterations of intracellular Ca2 + constitutes a very valuable therapeutic target. (Vallejo-Illarramendi et al. Expert Rev. Molec. Med. 2014, 16, e16, doi: 10.1017 / erm. 2014.17 "Dysregulation of calcium homeostasis in muscular dystrophies").
    High baseline levels of intracellular Ca2 + lead to the activation of calpain, protein degradation, the opening of mitochondrial permeability transition pores (PTPm) and, finally, the death of muscle fiber due to necrosis. The increase in intracellular Ca2 + levels is a complex process that involves Ca2 + flows through the sarcolemma, calcium losses from the sarcoplasmic reticulum (RS) and abnormal levels of Ca2 + in the RS. In mdx mice, an animal model of Duchenne muscular dystrophy, the abnormal S-nitrosylation of ryanodine receptor Ryr1 and RyR2 cysteine residues leads to the dissociation of calstabine from the protein complex, which produces unstable channels that lose calcium in the state of rest. This nitrosylation appears to be caused by a deregulation of nitric oxide that causes nitrosative and oxidative stress in the muscles of mdx dystrophic mice (Dudley et al., Am. J. Pathol. 2006; Bellinger et al., Nat Med. 2009, 15, 325 -330; Fauconier et al., Proc. Natl. Acad. Sci. USA 2010). In addition, induced microdistrophin expression in dystrophin-deficient myotubes reverses the increase in calcium release dependent on inositol 1,4,5-triphosphate receptors (IP3R) at control levels, suggesting the involvement of IP3R in the alteration of Ca2 + homeostasis in DMD.
    Immunoprecipitation assays have shown that calpain 3 (CAPN3) interacts with calsecuestrin (CSQ), a protein that participates in calcium homeostasis. In CAPN3 knockout mice (C3KO, Capn3 - / -), the reduction in RyR1 levels is accompanied by a reduction in the release of Ca2 + from RS. In turn, it has been seen that after the release of calcium from RS in Capn3 - / - mice, the reuptake of Ca2 + towards RS occurs more slowly, although the basis of this finding needs to be studied in more depth. However, it has been observed that the loss of CAPN3 proteolytic activity in Capn3 cs / cs mice does not affect calcium homeostasis, indicating that the structural function of CAPN3 is key to the maintenance of Ca2 + homeostasis .


    Myotonic dystrophy has been associated with the deregulation of the alternative connection of the CACNA1S gene that encodes the alpha 1S subunit of the DHPR dihydropyridine receptor, an essential voltage sensor in the excitation-contraction coupling (E-C). In DM1 and DM2, the omission of exon 29 of CACNA1S is greater, which increases the channel conductance and voltage sensitivity.
    Several disorders and diseases are associated with congenital or acquired modifications of the RyR1 protein (Kushmir et al. Recent Pat Biotechnol. 2012, 6, 157-166 "Ryanodine receptor patents"). Such disorders and diseases include conditions of the skeletal muscle, heart and nervous system. More specifically they include, but are not limited to, congenital myopathies, muscular dystrophies, sarcopenia, skeletal muscle fatigue, acquired muscle weakness or atrophy, malignant hyperthermia, irregular heart rhythm disorders and diseases associated with exercise, congestive heart failure, hypertrophic cardiomyopathy, Alzheimer's disease and memory loss associated with age. In all these conditions, it has been suggested that elevations of intracellular Ca2 + concentrations ([Ca2 +] i) at rest contribute directly to the toxicity of the cell (myofiber, cardiomyocyte, neuron or glial cell), its deterioration and simultaneous activation of Ca2 + dependent proteases.
    There are two main types of ryanodine receptors related to calcium channels in muscle fibers: RyR1 located in the skeletal muscle, and RyR2 located in the heart muscle. Each calcium channel is formed by a tetramer of RyR protein, each RyR monomer being able to interact with a casltabine protein. RyR1 binds to FKBP12 (calstabine1) and RyR2 binds to FKBP12.6 (calstabine2). Both in the heart and in skeletal muscle it has been seen that abnormal dissociation of calstabins and RyR channels by the progressive nitrosylation of the RyR channels, causes an increase in the release of Ca2 + from the sarcoplasmic reticulum to the intracellular cytoplasm, reducing the yield Muscle during contraction and activating, in the long term, muscular dysfunction (Bellinger et al., Nat. Med. 2009, 15, 325-330; Fauconier et al., Proc. Natl. Acad. Sci. USA. 2010, 107 , 1559-1564).
    Some low molecular weight compounds that show therapeutic activity in damaged muscle tissues are known in the state of the art. For example, the utility of 4- [3- (4-benzylpiperidinyl) -propanoyl] -2,3,4,5-tetrahydro-1,4-benzothiazepine (JTV-519) to prevent muscle necrosis has been demonstrated Cardiac and myocardial infarction. At concentrations of 10–6 M, compound JTV-519 inhibits in vitro myocardial necrosis induced by adrenaline and


    Caffeine in the left ventricle of a rat heart without affecting heart rate or left ventricular pressure (Kaneko, N. et al. WO92 / 12148A1 "Preparation of 4 - [(4benzylpiperidinyl) -alkanoyl] -2,3,4,5 -tetrahydro-1,4-benzothiazepine derivatives for inhibiting the kinetic cell death of cardiac muscles without inhibiting cardiac functions ”). Compound 5 JTV-519 acts on the ryanodine RyR2 receptor associated with calstabine2 (FKBP12.6), increasing the affinity of FKBP12.6 for the phosphorylated RyR2 receptor via PKA kinase, and also for the mutant RyR2 receptor which, otherwise mode, they have low affinity or do not bind to FKBP12.6. This JTV-519 action repairs the Ca2 + ion leak in RyR2 (Marks, A. R. et al US2004 / 229781A1 “Compounds and methods for treating and preventing exercise-induced
    10 cardiac arrhythmias ”).
    It is also known that pharmaceutically active compositions containing compound S107, or other structurally related tetrahydrobenzothiazepines, are effective in treating or preventing disorders or diseases related to RyR2 receptors that regulate the functioning of calcium channels in heart cells (Marks, AR et to US2006 / 194767A1 “Benzothiazepines as novel agents for preventing and treating disorders involving modulation of the RyR receptors and their preparation and pharmaceutical compositions”; see also: Mei, Y. et al. PLoS One. 2013, 8: e54208 “Stabilization of the skeletal muscle ryanodine ion channel-FKBP12 complex by the 1,4-benzothiazepine 20 derivative S107 ”) receiver. It is also known that the same family of compounds is active as a stabilizer of the RyR1-calstabine1 interaction, reducing muscle fatigue (Marks, AR et al. WO2008 / 060332A2 "Methods using tetrahydrobenzothiazepine compounds for treating or reducing muscle fatigue") and to treat sarcopenia (Marks, AR et al WO2012 / 019071A1 “Methods and compositions comprising benzazepine derivatives for preventing and treating
    25 sarcopenia ”).
    Some carvedilol derivatives have been described as assistants of the normalization of intracellular calcium homeostasis by action on RyR2 receptors, thus testing


    a beneficial effect in cardiac therapy (Chen, S. et al. US2007 / 025489A1 "Preparation of carbazoles as ryanodine receptor type 2 (RyR2) antagonists for treatment of cardiac conditions").
    Finally, it is known that the Ryom3 ryanodine receptor isomorph regulates intracellular calcium homeostasis in the brain or other neuronal tissues, and its modulation using 5 benzothiazepines has been claimed to treat some neuronal disorders (Marks, AR et al. WO2012 / 037105A1 " Methods and compositions comprising benzazepine derivatives for treating or preventing stress-induced neuronal disorders and diseases ”). In addition, both RyR1 and RyR2 are expressed in the brain, and there is evidence that calcium regulation and nitrooxidative stress are involved in various brain disorders and neuronal death (Kakizawa et
    10 al., EMBO J. 2012, 31, 417-428; Liu X et al., Cell. 2012, 150, 1055-1067).
    All these antecedents would show that the development of compounds that allow to regulate or modulate RyR receptors, regulating intracellular calcium levels, would provide an useful alternative for the treatment of muscular disorders, as well as in heart and neurodegenerative diseases. Summary of the invention
    The authors of the present invention have developed new compounds derived from triazole, in particular 4 - [(phenylthio) alkyl] -1H-1,2,3-triazoles, suitable for modulating RyR receptors that regulate calcium function in animal cells or human In the invention, said compounds
    20 are designated as "AHK".
    As evidenced in the experimental part, the compounds according to the present invention have the capacity to modulate intracellular calcium homeostasis in muscular dystrophic fibers, reversing the observed increases in intracellular calcium. In addition, said compounds have a modulating effect on RyR, their ability having been demonstrated
    25 to recover the RyR1-calstabine interaction on healthy human myotubes subjected to nitro-oxidative stress.
    On the other hand, in vivo tests have shown that these compounds also improve the grip strength in dystrophic mice, as well as normalize overexpressed dystrophic genes and reduce dystrophic histopathological markers.


    Therefore, a first aspect of the invention is related to a 1,2,3-substituted 1,4,3 substituted triazole compound of Formula (I): 5 (I)
    in which:
    R1 is a biradical C1-C4 alkylene optionally substituted by one or more substituents independently selected from the group consisting of C1-4 alkyl, C6-10 aryl, F, Cl, CN and 10 NO2;
    R2 is a C1-C6 biradical alkylene, in which 1, 2 or 3 -CH2- groups may be optionally replaced by groups selected from –O– and –S–; and wherein the biradical C1-C6 alkylene may be optionally substituted with one or more groups independently selected from C1-4 alkyl, allyl, propargyl, hydroxymethyl, 1-hydroxyethyl, 2
    15-hydroxyethyl, 3-aminopropyl, 4-aminobutyl, 3-guanidylpropyl, 3-indolylmethyl, C6-10 aryl, benzyl, 4-hydroxybenzyl, C6-10 heteroaryl, F, Cl, OH, O (C1-4 alkyl), CN , NO2, CO (C14 alkyl), CO2 (C1-4 alkyl), -CONH (C1-4 alkyl), -CON (C1-4 alkyl) 2;
    R3 is a group independently selected from H, C1-4 alkyl, C6-10 aryl, F, Cl, Br, I;
    m is selected from 0, 1, 2, 3, 4;
    20 n is selected from 0, 1, 2;
    X is independently selected from the group consisting of OH, O (C1-4 alkyl), O (C6-10 aryl), OCF3, S (C1-4 alkyl), S (C6-10 aryl), C1-6 alkyl , CF3, NHC (O) (C1-4 alkyl) and halogen;
    or two X groups may represent a birradical methylenedioxy, ethylenedioxy or propylenedioxy; and
    Y is selected from the group consisting of -OH, -CO2H, -CO2 (C1-4 alkyl), -CO2 (allyl),
    CO2 (benzyl), -SO3H, -NH2, -NH (C1-4 alkyl), -N (C1-4 alkyl) 2, N (C1-4 alkyl) 3 and N (heterocyclyl or heteroaryl), wherein said heterocyclyl or heteroaryl is found


    optionally substituted by a C1-4 alkyl group and where the N atom is part of the heterocyclyl or heteroaryl;
    or a stereoisomer, pharmaceutically acceptable salt, solvate or complex thereof, or an isotopically labeled derivative or a prodrug thereof.
    Another aspect of the invention comprises a method of synthesis of the compounds of Formula (I), which comprises:
    a) reacting an alkyne of Formula (II) with an azide of Formula (III),
    (O) n S R1 R3
    N3 R2Y
    (II) (III)
    10 optionally in the presence of a copper catalyst and optionally in the presence of a base, to give a compound of formula (I) as previously defined,
    where:
    the groups R1, R2, R3, m, n and X in the compounds of formulas (II) - (III) are as defined above, and
    Y is a group as defined above, optionally protected with a carboxyl protecting group, a hydroxyl protecting group or an amino protecting group;
    b) when n is 0 in the compound of formula (I) obtained in step a), optionally treating said compound of formula (I) with an oxidizing agent to give a compound
    20 of formula (I) wherein n is 1 or 2, R1, R2, R3, and X are as previously defined, and Y is a group as defined previously optionally protected with a carboxyl protecting group, a protecting group of hydroxyl or an amino protecting group; Y
    c) when the compound of formula (I) obtained in step a) or b) has a group Y
    Protected with a protective group, remove said protective group to give a compound of formula (I) wherein R1, R2, R3, m, n, X and Y are as previously defined.


    Another aspect of the invention is related to pharmaceutical compositions containing a compound of Formula (I), defined as indicated above, together with one or more excipients.
    or pharmaceutically acceptable vehicles.
    A further aspect of the present invention relates to the use of a compound of formula (I) as defined above for the preparation of a medicament.
    A final aspect of the invention is directed to the use of a compound of formula (I) as defined above in the preparation of a medicament for the treatment and / or prevention of disorders or diseases related to the deregulation of calcium concentration. intracellular or with RyR receptor dysfunction, in particular disorders or diseases of the
    10 skeletal muscle, disorders or heart disease and disorders or diseases of the nervous system. Brief description of the figures
    Figure 1 shows the toxicity curves of compounds AHK1, AHK2 and S-107 on 15 human myotubes after 24 h of incubation using the Citotox 96 colorimetric assay.
    Figure 2 shows the in vitro effect of compounds AHK1 and AHK2 on intracellular calcium levels at rest in mouse muscle fibers:
    A) Image of isolated fiber of the short digital flexor muscle that shows the characteristic striation pattern. B) Representative images of fibers [control fibers (CTRL) and dystrophic fibers
    20 (MDX)] loaded with fura-2AM after performing the background correction necessary to measure intracellular calcium levels. C) Histogram showing the basal levels of intracellular calcium in the different fiber groups. The number of fibers analyzed (n) is shown in the histogram (Kruskal-Wallis and U Mann-Whitney, * p <0.05).
    Figure 3 shows the in vitro effect of compounds AHK1 and AHK2 on the interaction RyR1-calstabine1 in healthy human myotube cultures exposed to peroxynitrite stress.
    Figure 4 shows the effect on grip strength in mdx dystrophic mice treated with compounds AHK1 and AHK2. * p <0.05; n = 10 control mice (CTRL); n = 11 untreated mdx mice (MDX); n = 10 mdx mice treated with AHK1 (MDX + AHK1); and n = 6 mdx mice treated with AHK2 (MDX + AHK2).


    Figure 5 shows the in vivo effect of compounds AHK1 and AHK2 on muscle degeneration / regeneration in dystrophic mdx mice:
    A) Cryostat sections representative of diaphragms of control and dystrophic mice in which the collagen IV labeled with a fluorescent antibody is observed, and the nuclei labeled with 5 4 ′, 6-diamidino-2-phenylindole (DAPI).
    B) Shows the number of central nuclei obtained in sections of control mice (CTRL), dystrophic mice (MDX) and dystrophic mice treated with AHK1 (MDX + AHK1) and AHK2 (MDX + AHK2). * P <0.05; n = 3 CTRL; n = 7 MDX ND; n = 7 MDX + AHK1 and n = 4 MDX + AHK2).
    Figure 6 shows the effect of treatment with AHK1 on the gene expression pattern of the anterior tibial muscle of mdx mice. Detailed description of the invention
    The present invention provides new triazole compounds that are capable of treating or preventing disorders or diseases associated with intracellular calcium dysregulation or dysfunction of RyR receptors.
    Thus, as previously mentioned, the first aspect of the present invention is directed to a compound of formula (I):
    (I)
    20 in which:
    R1 is a biradical C1-C4 alkylene optionally substituted by one or more substituents independently selected from the group consisting of C1-4 alkyl, C6-10 aryl, F, Cl, CN and NO2;
    R2 is a C1-C6 biradical alkylene, in which 1, 2 or 3 -CH2 groups may optionally be
    25 replaced by groups selected between –O– and –S–; and wherein the biradical C1-C6 alkylene may be optionally substituted with one or more groups independently selected from C1-4 alkyl, allyl, propargyl, hydroxymethyl, 1-hydroxyethyl, 2


    hydroxyethyl, 3-aminopropyl, 4-aminobutyl, 3-guanidylpropyl, 3-indolylmethyl, C6-10 aryl, benzyl, 4-hydroxybenzyl, C6-10 heteroaryl, F, Cl, OH, O (C1-4 alkyl), CN, NO2, CO (C14 alkyl), CO2 (C1-4 alkyl), -CONH (C1-4 alkyl), -CON (C1-4 alkyl) 2;
    R3 is a group independently selected from H, C1-4 alkyl, C6-10 aryl, F, Cl, Br, I;
    5 m is selected from 0, 1, 2, 3, 4;
    n is selected from 0, 1, 2;
    X is independently selected from the group consisting of OH, O (C1-4 alkyl), O (C6-10 aryl), OCF3, S (C1-4 alkyl), S (C6-10 aryl), C1-6 alkyl , CF3, NHC (O) (C1-4 alkyl) and halogen;
    or two X groups may represent a birradical methylenedioxy, ethylenedioxy or propylenedioxy; and
    10 Y is selected from the group consisting of -OH, -CO2H, -CO2 (C1-4 alkyl), -CO2 (allyl), CO2 (benzyl), -SO3H, -NH2, -NH (C1-4 alkyl), -N (C1-4 alkyl) 2 and N (C1-4 alkyl) 3 and N (heterocyclyl or heteroaryl), wherein said heterocyclyl or heteroaryl is optionally substituted by a C1-4 alkyl group and where the N atom is part of the heterocyclyl or heteroaryl;
    15 or a pharmaceutically acceptable stereoisomer, salt, solvate or complex thereof, or an isotopically labeled derivative thereof, or a prodrug thereof.
    In the context of the present invention, the following terms found in the compounds of formula (I) have the meaning indicated below:
    The term "birradical alkylene" refers to a birradical formed by a linear chain or
    20 hydrocarbon branched consisting of carbon and hydrogen atoms, which has no unsaturation and is attached at its ends to the rest of the molecule through simple bonds, such as, for example, methylene, ethylene, propylene, butylene, etc. The reference to biradical C1-C4 alkylene refers to when said birradical has between 1 and 4 carbon atoms, while the mention to birradical C1-C6 alkylene refers to when said
    Birradical has between 1 and 6 carbon atoms. The biradical alkylene may be substituted as specified in the definitions of the substituents R1 and R2 in the compound of formula (I).
    The term "alkyl" refers to a radical formed by a linear or branched hydrocarbon chain consisting of carbon and hydrogen atoms, which does not contain any saturation and is linked to the rest of the molecule by a single bond, for example


    methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, n-pentyl, etc. Mention to C1-C4 alkyl refers to when said radical has between 1 and 4 carbon atoms.
    The term "C6-C10 aryl" refers to a radical formed by an aromatic ring of between 6 and 10 members consisting of carbon and hydrogen atoms, preferably a phenyl radical.
    The term "C6-C10 heteroaryl" refers to a radical formed by an aromatic ring of between 6 and 10 members consisting of carbon and hydrogen atoms and one or more heteroatoms selected from O, N and S.
    The term "-N (heterocyclyl)" refers to a radical consisting of a cycle of between 5 and 7 members consisting of carbon and hydrogen atoms and one or more heteroatoms selected from O, N and S, at least one of them being N and this being directly attached to the radical R2.
    The term "-N (heteroaryl)" refers to a radical formed by an aromatic ring of between 6 and 10 members consisting of carbon and hydrogen atoms and one or more heteroatoms selected from O, N and S, at least one of they N and being directly attached to the radical R2.
    The term "allyl" refers to a radical of the formula -CH2-CH = CH2.
    The term "halogen" refers to F, Cl, Br or I.
    The term "isotopically labeled derivative" refers to a compound of formula (I) in which at least one of its atoms is isotopically enriched. For example, compounds of formula (I) in which a hydrogen is replaced by a deuterium or tritium, a carbon is replaced by a 13C or 14C enriched atom, or a nitrogen is replaced by a 15N enriched atom, they are within the scope of this invention.
    The term "pharmaceutically acceptable salts or solvates" refers to any pharmaceutically acceptable salt, ester, solvate, or any other compound that, in its administration to the recipient, is capable of providing (directly or indirectly) a compound of formula (I) such and as described herein. Salt preparation can be carried out by methods known in the state of the art.
    For example, pharmaceutically acceptable salts of the compounds provided herein are synthesized from the compound described above containing


    a basic or acid unit by conventional chemical methods. In general, such salts are prepared, for example, by reacting the free acidic or basic forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of both. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. Examples of acid addition salts include addition salts of mineral acids such as, for example, hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate, and organic acid addition salts such as, for example, acetate, maleate, fumarate , citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulfonate and p-toluenesulfonate. Examples of alkaline addition salts include inorganic salts such as, for example, sodium, potassium, calcium, ammonium, magnesium, aluminum and lithium, and organic alkaline salts such as, for example, ethylenediamine, ethanolamine, N, N-dialkylene ethanolamine, glucamine and salts. basic amino acids
    Solvates refer to a salt of the compound of formula (I) in which the molecules of a pharmaceutically suitable solvent are incorporated into the crystalline network. Solvation methods in general are known in the state of the art. Examples of pharmaceutically suitable solvents are ethanol, water and the like. In a particular embodiment the solvate is a hydrate.
    The compounds of formula (I) or their salts or solvates are preferably in pharmaceutically acceptable form or in substantially pure form. As a pharmaceutically acceptable form it is understood, inter alia, that they have a pharmaceutically acceptable level of purity, excluding normal pharmaceutical additives such as diluents and excipients, and not including any material considered toxic at normal dosage levels. The purity levels for the drug are preferably above 50%, more preferably above 70%, and even more preferably above 90%. In a preferred embodiment it is above 95% of the compound of formula (I), or of its salts or solvates.
    The compounds of the present invention represented by the formula (I) described above may include any stereoisomer depending on the presence of chiral centers, including enantiomers and diastereoisomers. The individual isomers, enantiomers or diastereomers and mixtures thereof are within the scope of the present invention.
    The term "prodrug" is used in its broadest sense and encompasses those derivatives that are converted in vivo into the compounds of the invention. Such derivatives include, depending


    of the functional groups present in the molecule and without limitation, esters, amino acid esters, phosphate esters, sulphonate esters of metal salts, carbamates and amides. Examples of methods for producing a prodrug of a given active compound are known to one skilled in the art and can be found, for example, in Krogsgaard-Larsen et al. "Textbook of Drugdesign and Discovery" Taylor & Francis (April 2002).
    In a variant (A) of the present invention, the radical R1 is -CH2-.
    In a preferred embodiment of said variant (A), R2 is a birradical C1-4 alkylene optionally substituted with one or two substituents independently selected from the group consisting of methyl, ethyl, propyl, isopropyl, allyl, propargyl, butyl, isobutyl, secbutyl , tert-butyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 3-aminopropyl, 4-aminobutyl, 3-guanidylpropyl, 3-indolylmethyl, phenyl, 1-naphthyl, 2-naphthyl, benzyl, 4-hydroxybenzyl, and C6 heteroaryl -10.
    More preferably, R2 is a birradical -CH2-or -CH2-CH2-, optionally substituted with one or two substituents independently selected from the group consisting of methyl, isopropyl, isobutyl and benzyl. Even more preferably, R2 is a birradical -CH2 optionally substituted with two methyl substituents or with a substituent selected from the group consisting of isopropyl, isobutyl and benzyl, or R2 is a birradical -CH2-CH2-. In a also preferred form, R2 is a birradical -CH2-or -CH2-CH2-.
    In another preferred embodiment of said variant (A), R3 is H.
    In another preferred embodiment of said variant (A), m is 1.
    In another preferred embodiment of said variant (A), n is 0.
    In another preferred embodiment of said variant (A), X is -O (C1-4 alkyl) or halogen, more preferably it is a methoxy group in meta or para position or chlorine.
    In another preferred embodiment of said variant (A), Y is selected from the group consisting of CO2H, CO2Me, NH2, -NHMe, -NMe2, -NMe3, -NHEt, -NEt2, -NEt3,
    or a pharmaceutically acceptable salt of said groups.


    More preferably, Y is selected from the group consisting of CO2H, CO2Me, NH2, -NHMe, -NMe2, -NMe3, -NHEt, -NEt2, -NEt3, pyrrolidin-1-yl, piperidin-1-yl, morpholin -4-yl, 4piperazin-1-yl, 4-methyl-piperazin-1-yl, pyridin-1-yl, more preferably CO2H and -NMe2, even more preferably Y is -CO2H or a pharmaceutically acceptable salt thereof.
    Within the variant (A), the compounds of formula (I) are selected from the group consisting of: 1-carboxymethyl-4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, 1- [2- (N, N-dimethylamino) ethyl] -4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, 1-carboxymethyl-4- [4- (methoxy) phenylthiomethyl] -1H -1,2,3-triazole, (S) -1- (1-carboxy-2-phenylethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, (R) - 1- (1-carboxy-2-phenylethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, (S) -1- (1-carboxy-3-methylbutyl) -4 - [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, (R) -1- (1-carboxy-3-methylbutyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1 , 2,3-triazole, (S) -1- (1-carboxy-2-methylpropyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, (R) -1- (1-carboxy-2-methylpropyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, 1- (1-carboxy-1-methyl ethyl) -4- [3- (methoxy ) phenylthiomethyl] -1H-1,2,3-triazole, 1- (2-hydroxyethyl) -4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, 1-methoxycarbonylmethyl-4- ( phenylsulfinylmethyl) -1H-1,2,3-triazole, and 1-carboxymethyl-4- [3- (methoxy) phenylsulfoni lmethyl] -1H-1,2,3-triazole, 1-carboxymethyl-4- [3- (chloro) phenylthiomethyl] -1H-1,2,3-triazole,
    or a salt thereof.In a variant (B) of the present invention, the radical R1 is -CH2-.In a preferred embodiment of said variant (B), R2 is a C1-4 biradical alkylene in which
    one or two -CH2-groups are replaced by -O-, and wherein said birradical C1-C4 alkylene is optionally substituted with one or two C1-C4 alkyl groups, preferably selected from methyl, ethyl, n-propyl, isopropyl, n -butyl and tert-butyl.
    More preferably, R2 is a birradical -CH2-or -CH2-CH2-, optionally substituted with


    one or two substituents independently selected from the group consisting of methyl, isopropyl and isobutyl. Even more preferably, R2 is a birradical -CH2-optionally substituted with two methyl substituents or with a substituent selected from the group consisting of isopropyl and isobutyl, or R2 is a birradical -CH2-CH2-.
    In another preferred embodiment of said variant (B), R3 is H. In another preferred embodiment of said variant (B), m is 1. In another preferred embodiment of said variant (B), n is 0. In another preferred embodiment of said variant (B), X is -O (C1-4 alkyl) or halogen,
    more preferably it is a methoxy group in meta or para or chlorine position. In another preferred embodiment of said variant (B), Y is selected from the group consisting of NH2, -NH (C1-C4 alkyl), -N (C1-C4 alkyl) 2, -N (C1-C4 alkyl )3,
    More preferably Y is selected from -NH2, -NHMe, -NMe2, -NMe3, -NHEt, -NEt2, -NEt3, pyrrolidin-1-yl, piperidin-1-yl, morpholin-4-yl, 4-piperazin-1 -yl, 4-methyl-piperazin-1-yl,
    15 pyridin-1-yl or a pharmaceutically acceptable salt of said groups. More preferably, Y is -NMe2, or a pharmaceutically acceptable salt thereof.
    In a variant (C) of the present invention, the radical R1 is -CH2-.
    In a preferred embodiment of said variant (C), R2 is a birradical C1-4 alkylene optionally substituted with one or two substituents independently selected from the
    A group consisting of methyl, ethyl, propyl, isopropyl, allyl, propargyl, butyl, isobutyl, secbutyl, tert-butyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 3-aminopropyl, 4-aminobutyl, 3-guanidylpropyl, 3 -indolylmethyl, phenyl, 1-naphthyl, 2-naphthyl, benzyl, 4-hydroxybenzyl, and C6-10 heteroaryl.
    More preferably, R2 is a birradical -CH2-or -CH2-CH2-, optionally substituted with
    One or two substituents independently selected from the group consisting of methyl, isopropyl, isobutyl and benzyl. Even more preferably, R2 is a birradical -CH2 optionally substituted with two methyl substituents or with a substituent selected from the group consisting of isopropyl, isobutyl and benzyl, or R2 is a birradical -CH2-CH2-. In a


    Also preferred, R2 is a birradical -CH2-or -CH2-CH2-.In another preferred embodiment of said variant (C), R3 is H.In another preferred embodiment of said variant (C), m is 1.In another preferred embodiment of said variant (C), n is 0.In another preferred embodiment of said variant (C), X is -O (C1-4 alkyl) or halogen,
    more preferably it is a methoxy group in meta or para or chlorine position.
    In another preferred embodiment of said variant (C), Y is selected from the group that
    consists of CO2H, CO2 (C1-C4 alkyl), NH2, -NH (C1-C4 alkyl), -N (C1-C4 alkyl) 2, -N (alkyl
    C1-C4) 3,
    or a pharmaceutically acceptable salt of said groups.
    More preferably Y is selected from -NH2, -NHMe, -NMe2, -NMe3, -NHEt, -NEt2, -NEt3, pyrrolidin-1-yl, piperidin-1-yl, morpholin-4-yl, 4-piperazin-1 -yl, 4-methyl-piperazin-1-yl, pyridin-1-yl or a pharmaceutically acceptable salt of said groups.
    More preferably, Y is selected from the group consisting of CO2H and -NMe2 or a pharmaceutically acceptable salt thereof.
    Method of obtaining the compounds of the invention
    The compounds of formula (I) of the present invention can be prepared by a process comprising reacting an alkyne of formula (II) with an azide of formula (III):
    (O) n S R1 R3
    N3 R2Y
    (II) (III)
    wherein R1, R2, R3, m, n and X in the compounds of formulas (II) - (III) are as defined for the compounds of formula (I), and where Y is a group as defined for the compounds of formula (I), optionally protected with a group


    carboxyl protector, a hydroxyl protecting group or an amino protecting group, depending on the nature of said group Y.
    This reaction can be carried out in the presence of a copper catalyst, such as, for example, copper (II) sulfate / sodium ascorbate, copper (I) iodide or copper (I) acetate.
    In addition, in a preferred embodiment, the reaction between the compound of formula (II) and the compound of formula (III) is carried out in the presence of a base, such as, for example, sodium acetate, diisopropylamine or triethylamine.
    In a particular embodiment, when n is 0, the compound of formula (I) obtained according to the above procedure can be treated with an oxidizing agent to give a compound of
    10 formula (I) in which n is 1 or 2.
    Examples of oxidizing agents include, for example, 3-chloroperbenzoic acid or tert-butyl hydroperoxide.
    In another particular embodiment, when the compound of formula (I) obtained according to the above procedure has a Y group protected with a protective group, said protective group 15 is removed to give a compound of formula (I) wherein R1, R2, R3 , m, n, X and Y are as defined for the compound of formula (I). The removal of the protective group can be performed following procedures commonly known to an expert in organic synthesis.
    Pharmaceutical compositions
    The present invention further provides pharmaceutical compositions comprising a compound of formula (I), or a pharmaceutically acceptable stereoisomer, salt, solvate or complex thereof, or an isotopically labeled derivative thereof, or a prodrug thereof; and one or more pharmaceutically acceptable excipients or vehicles.
    The pharmaceutically acceptable vehicle must be acceptable in the sense of being compatible with
    25 the other ingredients of the composition and not harmful to the recipient thereof. Said pharmaceutically acceptable carrier can be selected from among organic and inorganic materials that are used in pharmaceutical formulations and which are incorporated as analgesic agents, pH regulators, binders, disintegrants, diluents, emulsifiers, fillers, glidants, solubilizers, stabilizers, suspending agents, tonicity agents and
    30 thickeners Pharmaceutical additives such as antioxidants, agents may also be added.


    aromatics, dyes, aroma enhancing agents, preservatives and sweeteners.
    Examples of pharmaceutically acceptable carriers include, among others, carboxymethyl cellulose, crystalline cellulose, glycerin, gum arabic, lactose, magnesium stearate, methyl cellulose, saline, sodium alginate, sucrose, starch, talc and water.
    The pharmaceutical formulations of the present invention are prepared by procedures well known in the pharmaceutical art. For example, the compounds of formula (I) are mixed with a pharmaceutically acceptable carrier or carrier, such as a suspension or solution. The choice of vehicle is determined by the solubility and chemical nature of the compounds, the route of administration chosen and the standard pharmaceutical practice.
    The administration of the compounds or compositions of the present invention to a human or animal subject may be by any known procedure including, without limitation, oral administration, sublingual or oral administration, parenteral administration, transdermal absorption, via nasal inhalation or instillation. , vaginal, rectal and intramuscular administration.
    In a particular embodiment, administration is carried out parenterally, such as by subcutaneous injection, intramuscular injection, intraperitoneal injection, intravenous injection. For parenteral administration, the compounds of the invention are combined with a sterile aqueous solution that is isotonic with the subject's blood. Such a formulation is prepared by dissolving the solid active ingredient in water containing
    20 physiologically compatible substances, such as sodium chloride, glycine and the like, and having a buffered pH compatible with physiological conditions. Said formulation is presented in single or multiple dose containers, such as ampoules or closed vials.
    In another particular embodiment, administration is performed orally. In that case, the formulation of the compounds of the invention can be presented as capsules,
    25 tablets, powders, granules, or as a suspension or solution. Said formulation may include conventional additives, such as lactose, mannitol, starch, etc .; binders, such as crystalline cellulose, cellulose derivatives, gum arabic, corn starch or gelatins; disintegrants, such as corn starch and potato starch or sodium carboxymethyl cellulose; lubricants, such as talc or magnesium stearate.


    Applications
    The compounds of the present invention have proved suitable for modulating RyR receptors that regulate the function of calcium in animal or human cells, so that they are capable of treating or preventing disorders or diseases associated with the deregulation of intracellular calcium caused primarily as a consequence of RyR receptor dysfunction.
    The term "deregulation of intracellular calcium" means an abnormal regulation of calcium levels and calcium fluxes in cells.
    Thus, an increase in the intracellular concentration of calcium (Ca2 +) under resting conditions contributes to the damage of toxic muscle cells (myofibers) and the simultaneous activation of Ca2 + dependent proteases, such as calpain. Since the activity of calpain is increased in necrotic muscle fibers of mdx mice and calpain dysfunction contributes to myodistrophy of the waist and extremities, the prevention of calcium-dependent protease activity by inhibiting intracellular elevations of Ca2 + makes it possible to avoid muscular atrophy and, therefore, treat diseases such as Duchenne's myodystrophy or Becker's myodistrophy.
    In vitro tests performed with the compounds of the invention have shown their ability to reverse intracellular calcium increases in muscle fibers, suggesting that these compounds perform this reversal through a mechanism that involves modulating RyR channels. In addition, said compounds have also shown the ability to reduce by half the number of overexpressed genes in mdx dystrophic muscle, some of which is related to the loss of skeletal muscle or atrophy.
    RyR receptors that regulate intracellular calcium function include RyR1, RyR2 and RyR3, as well as a RyR protein or a RyR analog. A RyR analog refers to a functional variant of RyR protein with biological activity that has a homology of 60% or higher in the amino acid sequence with the RyR protein.
    In the context of the present invention, "biological activity of RyR" means the activity of the protein or peptide that demonstrates an ability to physically associate with, or bind to, FKBP 12 (calstabine-1) in the case of RyR1 and RyR3, and FKBP12.6 (calstabine-2) in the case of RyR2 under the conditions of the tests described herein.
    The compounds of the present invention can be used to limit or prevent


    decrease in the level of FKBP (calstabine) bound to RyR in the cells of a subject. Based on what is described in the previous paragraph, FKBP linked to RyR refers to FKBP12 linked to RyR1 (calstabine-1), FKBP 12.6 linked to RyR2 (calstabine-2) and FKBP12 linked to RyR3 (casltabine-1).
    A decrease in the level of FKBP linked to RyR in the cells of a subject is limited or avoided when said decrease is, in any case, stopped, hindered, impeded, obstructed.
    or reduced by the administration of the compounds of the invention, so that the level of FKBP bound to RyR in the cells of a subject is higher than it would otherwise be in the absence of the administered compound.
    The level of FKBP bound to RyR in a subject is detected by standard tests or techniques known to one skilled in the art, such as immunological techniques, hybridization analysis, immunoprecipitation, Western blot analysis, fluorescence imaging techniques and / or detection. of radiation, as well as any other as disclosed in the experimental part of this document.
    In a particular embodiment, the decrease in the level of FKBP (calstabine) bound to RyR occurs as a result of subjecting the cells of a subject to nitro-oxidative stress. In fact, experimental tests carried out with the compounds of the invention have shown that they allow minimizing the degenerative effects of nitro-oxidative stress on healthy human myotubes through an increase in the affinity of the RyR1calstabine-1 interaction.
    Consequently, it has been shown that the compounds of the invention avoid disorders or conditions that involve the modulation of RyR receptors or the increase of intracellular calcium, thus allowing the levels thereof to be regularized. Examples of such disorders or conditions include skeletal muscle disorders and diseases (related to RyR1 modulation), cardiac disorders and diseases (related to RyR2 modulation) and nervous system disorders and diseases (related to RyR1 modulation, RyR2 or RyR3).
    Thus, a further aspect of the present invention relates to the use of the compounds of formula (I), or a pharmaceutically acceptable stereoisomer, salt, solvate or complex thereof, or an isotopically labeled derivative thereof, or a prodrug thereof. , in the preparation of a medicament aimed at the treatment and / or prevention of skeletal muscle disorders and diseases, cardiac disorders and diseases and disorders and


    diseases of the nervous system.
    In a particular embodiment, skeletal muscle disorders and diseases are selected from muscular dystrophies, congenital myopathies, metabolic myopathies and muscular atrophy. Preferably, said skeletal muscle disorder or disease is Duchenne muscular dystrophy or Becker muscular dystrophy.
    In another particular embodiment, cardiac disorders and diseases are selected from heart failure, cardiac ischemia, cardiac arrhythmias and cardiomyopathies.
    In another particular embodiment, disorders and diseases of the nervous system are selected from brain accident, Alzheimer's disease, frontotemporal dementia and cognitive impairment.
    In a preferred embodiment, the compounds according to variant (A) of the present invention are those used in the preparation of a medicament for the treatment of skeletal muscle disorders and diseases, as well as for the treatment of cardiac disorders and diseases, such as those described. previously.
    In another preferred embodiment, the compounds according to variant (B) of the present invention are those used in the preparation of a medicament for the treatment of disorders and diseases of the nervous system as described above.
    A further aspect of the present invention relates to a compound of formula (I), or a pharmaceutically acceptable stereoisomer, salt, solvate or complex thereof, or an isotopically labeled derivative thereof, or a prodrug thereof, for use in the treatment and / or prevention of disorders and diseases of skeletal muscle, disorders and heart disease and disorders and diseases of the nervous system.
    Another aspect of the invention is directed to a method for the treatment and / or prevention of skeletal muscle disorders and diseases, disorders and heart diseases and disorders and diseases of the nervous system, which comprises administration to a patient in need of an amount Therapeutically effective of a compound of formula (I), or a pharmaceutically acceptable stereoisomer, salt, solvate or complex thereof, or an isotopically labeled derivative thereof, or a prodrug thereof.
    By "therapeutically effective" amount should be understood as sufficient to achieve beneficial or desired results, the preventive and / or therapeutic response being, avoiding or


    substantially mitigating unwanted side effects.
    In a particular embodiment, the compounds of the present invention are administered to a subject in an amount effective to modulate abnormal intracellular calcium concentrations. This amount can easily be determined by an expert in the area using known procedures. For example, the release of intracellular calcium through RyR channels can be quantified using calcium-sensitive fluorescent dyes, such as Fluo-3 or Fura-2, and monitoring calcium-dependent fluorescence signals with a photomultiplier tube and software adequate (Brillantes, et al. Cell, 1994, 77, 513-523, "Stabilization of calcium release channel (ryanodine receptor) function by FK506-binding protein"; Gillo, et al. Blood, 1993, 81, 783-792) .
    The concentration of the compounds of the invention in serum of a human or animal subject can be determined following methods known in the art (Thebis, M. et al. Drug Test. Analysis 2009, 1, 32-42 "Screening for the calstabin-ryanodine complex stabilizers receiver JTV-519 and S-107 in doping control analysis ”). The administered amount of compounds of Formula (I) that is effective in limiting or preventing abnormal levels of intracellular calcium will depend on the relative efficacy of the compound chosen, the severity of the disorder treated and the weight of the affected. However, typically the compounds will be administered one or more times a day, for example 1, 2, 3 or 4 times daily, with total typical daily doses in the range of approximately, between 1 mg / kg / day and 100 mg / kg / day, and more preferably between 10 mg / kg / day and 40 mg / kg / day, or an amount sufficient to achieve serum levels between approximately 1 ng / mL and 500 ng / mL.
    The compounds of Formula (I) can be used alone, in combination with each other, or in combination with other drugs that have therapeutic activity including, but not limited to, mRNA exon skip enhancers, gene transcription modulators, diuretics, anticoagulants, agents platelets, antiarrhythmics, inotropic agents, chronotropic agents, α and β blockers, angiotensin inhibitors and vasodilators.
    Such drugs may be part of the same composition, or be provided as a separate composition, for administration at the same time or at a different time.
    The present invention also includes an in vitro method for determining the ability of a compound to modulate intracellular calcium levels and prevent the dissociation of calstabine from the RyR protein complex, wherein said method comprises: (a) obtaining or generating a culture


    cell containing RyR receptors; (b) contacting the cells with the compound to be tested; (c) exposing the cells to one or more known conditions that alter intracellular calcium regulation or generate post-translational modifications in the RyR receptor; (d) determine if said compound modulates intracellular calcium levels; and (e) determining whether said compound limits or prevents the dissociation of calstabine from the RyR protein complex.
    In a particular embodiment, the conditions that alter intracellular calcium regulation or generate post-translational modifications in the RyR receptor are oxidative stress or nitrosative stress.
    The present invention also contemplates a method of diagnosing a disorder or disease, wherein said method comprises:
    - obtain a sample of tissue or cells from a subject that contains RyR receptors; -include the tissue sample or cells obtained in step a) with a compound of formula (I), and -determine:
    (to) if there is an increase in the interaction between RyR and calstabine with respect to
    to the interaction between RyR and calstabine in cells or control tissue; or
    (b) if there is a decrease in intracellular calcium levels compared to an absence of such decrease in control cells or tissue;
    where an increase in the interaction between RyR and calstabine in (a) or a decrease in intracellular calcium levels in (b), indicates the presence of a disorder or disease in the subject.
    In a particular embodiment, the compound of formula (I) employed in the diagnostic method is a compound according to variant (C) of the present invention.
    In another particular embodiment, the tissue sample is a muscle tissue sample.
    In a particular embodiment, when RyR is RyR1, the disorder or disease to be diagnosed is a skeletal muscle disorder or disease.
    In a particular embodiment, when RyR is RyR2, the disorder or disease to be diagnosed is a heart disease or disorder.
    In a particular embodiment, when RyR is RyR1, RyR2 or RyR3, the disorder or disease a


    diagnose is a disorder or disease of the nervous system.
    The measures of increase in the RyR and calstabine interaction and of decrease in intracellular calcium levels can be carried out by techniques known by an expert, such as immunoprecipitation, in situ proximity ligation assays (PLA) and calcium images at
    5 real time using fluorescent probes. Examples
    Acronyms of compounds, reagents, solvents or the techniques employed are defined as follows:
    - AHK1: is a compound according to Formula (I) that contains the groups: R1 = R2 = –CH2–; 10 R3 = H; m = 1; n = 0; X = 3-MeO; Y = CO2H, -AHK2: is a compound according to Formula (I) that contains the groups: R1 = –CH2–; R2 = –CH2CH2–; R3 = H; m = 1; n = 0; X = 4-MeO; Y = NMe2, -S-107: compound used for comparative purposes in biological tests performed whose structure is found in the background of this document.
    15 -tBuOH: tert-butanol, -DAPI: 4 ’, 6-Diamino-2-phenylindole, -ESI: Electrospray ionization, -EtOAc: Ethyl acetate, -HRMS: High resolution mass spectrometry,
    20 -IR: Infrared spectroscopy, -mdx: Animal model of Duchenne muscular dystrophy related to the X chromosome, -Pf: Melting point, -RMN: Nuclear magnetic resonance, -SIN1: 3-Morpholino-sidnonimine, peroxynitrite generating agent and Nitric oxide.
    -THF: Tetrahydrofuran, -TBTA: Tris [(1-benzyl-1H-1,2,3-triazol-4-yl) methyl] amine,
    The following examples are provided for illustrative purposes, and do not imply a limitation of the present invention.
    EXAMPLE 1: 1-Methoxycarbonylmethyl-4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.


    A solution of 1- (4-methoxyphenylthio) -2-propyne (2 mmol, 356 mg), methyl azidoacetate (2.00 mmol, 230 mg) and TBTA (catalyst) in THF / tBuOH (1: 1) was prepared , 4 mL) under nitrogen atmosphere. Then, two degassed aqueous solutions of CuSO4 (0.4 mmol, 63 mg) in H2O (1 mL) and sodium ascorbate (0.8 mmol, 158 mg) in H2O (1 mL) were added and the mixture was stirred at room temperature overnight. Upon completion of the reaction, the solvent was evaporated, a 28% solution of aqueous ammonia (10 mL) was added and the product was extracted with CH2Cl2 (3 x 20 mL). The combined organic extracts were dried (MgSO4) and the solvent was evaporated under reduced pressure. The product was purified by column chromatography (silica gel; EtOAc / hexane 1: 3). Rdto: 477 mg (78%). Yellowish oil. IR (cm-1): 2953, 2837 (C-H), 1750 (C = O), 1219, 1174 (triazole). 1H NMR (500 MHz, CDCl3): δ 7.38 (s, 1H), 7.30 (d, J = 8.7 Hz, 2H), 6.81 (d, J = 8.7 Hz, 2H) , 5.09 (s, 3H), 4.12 (s, 2H), 3.78 (s, 3H), 3.77 (s, 3H). 13C NMR (125 MHz, CDCl3): δ 166.7, 159.4, 145.8, 134.0, 125.4, 123.5, 114.7, 55.4, 53.1, 50.8, 30.9 HRMS (ESI +, m / z) calculated for C13H16N3O3S:
    15 294.0912; Found: 294.0916.
    EXAMPLE 2: 1-Methoxycarbonylmethyl-4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of Example 1 was followed, starting from 1- (3-methoxyphenylthio) -2-propyne (2.00
    20 mmol, 356 mg) and methyl azidoacetate (2.00 mmol, 230 mg). Rdto: 537 mg (88%). Yellowish oil. IR (cm – 1): 2953, 2837 (C-H), 1749 (C = O), 1220, 1180 (triazole). 1 H NMR (500 MHz, CDCl 3): δ 7.51 (s, 1 H), 7.18 (t, J = 8.0 Hz, 1 H), 6.92 (d, J = 7.7 Hz, 1 H) , 6.90-6.87 (m, 1H), 6.73 (dd, J = 8.2, 1.8 Hz, 1H) 5.11 (s, 2H), 4.26 (s, 2H) , 3.78 (s, 3H), 3.77 (s, 3H). 13C NMR (125 MHz, CDCl3): δ 166.7, 159.9, 145.7, 136.8, 129.9, 123.6, 121.5, 114.6, 112.5, 55.4, 53.1, 25 50.8, 28.7. HRMS (ESI +, m / z) calculated for C13H16N3O3S: 294.0912; Found: 294.0917.


    EXAMPLE 3: (S) -1- (1-Methoxycarbonyl-2-phenylethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of Example 1 was followed, starting from 1- (3-methoxyphenylthio) -2-propyne (2.00
    5 mmol, 384 mg) and the methyl ester of (S) -2-azido-3-phenylpropanoic acid (2.00 mmol, 358 mg). Rdto: 668 mg (87%). Yellowish oil. [α] D 25 = -56.1 ° (c. 1.01 g / 100 mL, CH2Cl2). IR (cm-1): 2952, 2836 (C-H), 1744 (C = O), 1228, 1172 (triazole). 1H NMR (400 MHz, CDCl3): δ 7.47 (s, 1H), 7.27-6.62 (m, 9H), 5.50 (dd, J = 8.5, 6.2 Hz, 1H ), 4.12 (q, J = 14.9 Hz, 2H), 3.75 (s, 3H), 3.73 (s, 3H), 3.46 (dd, J = 14.0, 5, 8 Hz, 1H), 3.36 (dd, J = 13.9, 9.2 Hz, 1H). 13C NMR (101
    10 MHz, CDCl3): δ 168.6, 159.9, 145.1, 136.8, 134.7, 129.9, 128.9, 127.6, 122.4, 121.5, 114.5 , 112.5, 64.3, 55.4, 53.2, 38.9, 28.7. HRMS (ESI +, m / z) calculated for C20H22N3O3S: 384.1382; Found: 384.1392.
    EXAMPLE 4: (R) -1- (1-Methoxycarbonyl-2-phenylethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of example 1 was followed, starting from 1- (3-methoxyphenylthio) -2-propyne (2.00 mmol, 384 mg) and the methyl ester of (R) -2-azido-3-phenylpropanoic acid (2, 00 mmol, 358 mg). Rdto: 653 mg (85%). Yellowish oil. [α] D 25 = + 39.4 ° (c. 2.71 g / 100 mL, CH2Cl2). HRMS (ESI +, m / z) calculated for C20H22N3O3S: 384.1382; Found: 384.1382. NMR data
    20 were identical to those in example 3.


    EXAMPLE 5: (S) -1- (1-Methoxycarbonyl-3-methyl-butyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3triazole.
    The procedure of Example 1 was followed, starting from 1- (3-methoxyphenylthio) -2-propyne (2.00
    5 mmol, 384 mg) and the methyl ester of (S) -2-azido-4-methylpentanoic acid (2.00 mmol, 342 mg). Rdto: 677.9 mg (97%). Yellowish oil. [α] D 25 = + 10.7 ° (c. 1.00 g / 100 mL, CH2Cl2). 1 H NMR (500 MHz, CDCl 3): δ 7.51 (s, 1 H), 7.16 (t, J = 8.0 Hz, 1 H), 6.90 (d, J = 7.7 Hz, 1 H) , 6.87 (s, 1H), 6.72 (dd, J = 8.2, 1.8 Hz, 1H), 5.38 (t, J = 8.0 Hz, 1H), 4.24 ( q, J = 14.8 Hz, 2H), 3.76 (s, 3H), 3.73 (s, 3H), 1.94 (t, J = 7.5 Hz, 2H), 1.19 ( dt, J = 13.4, 6.7 Hz, 1H), 0.90 (d, J = 6.5 Hz,
    10 3H), 0.84 (d, J = 6.6 Hz, 3H). 13C NMR (125 MHz, CDCl3): δ 169.8, 159.9, 145.3, 136.6, 129.8, 122.1, 115.1, 112.7, 61.1, 55.3, 53.1, 41.4, 29.1, 24.7, 22.7, 21.3.
    EXAMPLE 6: (R) -1- (1-Methoxycarbonyl-3-methyl-butyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3triazole.
    The procedure of Example 1 was followed, starting from 1- (3-methoxyphenylthio) -2-propyne (2.00 mmol, 384 mg) and the methyl ester of (R) -2-azido-4-methylpentanoic acid (2.00 mmol , 342 mg). Rdto: 653 mg (93%). Yellowish oil. [α] D 25 = -12.1 ° (c. 1.03 g / 100 mL, CH2Cl2). The NMR data were identical to those in example 5.
    EXAMPLE 7: (S) -1- (1-Methoxycarbonyl-2-methyl-propyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3triazole.


    The procedure of Example 1 was followed, starting from 1- (3-methoxyphenylthio) -2-propyne (2.00 mmol, 384 mg) and the methyl ester of (S) -2-azido-3-methylbutanoic acid (2, 00 mmol, 314 mg). Rdto: 616 mg (92%). Yellowish oil. [α] D 25 = + 21.5 ° (c. 1.03 g / 100 mL, CH2Cl2). 1 H NMR
    5 (500 MHz, CDCl3): δ 7.63 (s, 1H), 7.16 (t, J = 7.9 Hz, 1H), 6.92 (d, J = 7.5 Hz, 1H), 6.88 (s, 1H), 6.72 (d, J = 6.8, 1H), 5.06 (d, J = 8.7 Hz, 1H), 4.24 (q, J = 14, 7 Hz, 2H), 3.76 (s, 6H), 2,442.30 (m, 1H), 0.96 (d, J = 6.6 Hz, 3H), 0.73 (d, J = 6, 6 Hz, 3H). 13C NMR (125 MHz, CDCl3): δ 168.8, 159.6, 144.7, 136.3, 129.5, 121.9, 115.0, 112.3, 68.5, 55.0, 52.5, 32.0, 28.8, 18.8, 18.1.
    EXAMPLE 8: (R) -1- (1-Methoxycarbonyl-2-methyl-propyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3triazole.
    The procedure of Example 1 was followed, starting from 1- (3-methoxyphenylthio) -2-propyne (2.00 mmol, 384 mg) and the methyl ester of (R) -2-azido-3-methylbutanoic acid (2, 00 mmol, 314 mg).
    15 Rdto: 626 mg (93%). Yellowish oil. [α] D 25 = -19.5 ° (c. 1.06 g / 100 mL, CH2Cl2). The NMR data were identical to those in example 7.
    EXAMPLE 9: 4- [3- (Methoxy) phenylthiomethyl] -1- (1-methyl-1-methoxycarbonylethyl) -1H-1,2,3-triazole.
    O o
    N SN
    N
    OR


    The procedure of Example 1 was followed, starting from 1- (3-methoxyphenylthio) -2-propyne (2.00 mmol, 384 mg) and methyl 2-azidoisobutyrate (2.00 mmol, 386 mg). Rdto: 258 mg (40%). Yellowish oil. 1 H NMR (400 MHz, CDCl 3): δ 7.44 (s, 1 H), 7.11 (t, J = 7.7 Hz, 1 H), 6.85 (d, J = 7.6 Hz, 1 H) , 6.82 (s, 1H), 6.67 (d, J = 7.6 Hz, 1H), 4.17 (s, 2H), 3.69 (s, 3H), 3.62 (s, 3H), 1.82 (s, 6H). 13C NMR (101 MHz, CDCl3) δ 171.6, 159.7, 144.3, 136.7, 129.6, 121.6, 121.0, 114.8, 112.3, 64.3, 55 , 1, 53.1, 28.8, 25.5.
    EXAMPLE 10: 1- (2-Hydroxyethyl) -4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of Example 1 was followed, starting from 1- (4-methoxyphenylthio) -2-propyne (2.00 mmol, 384 mg) and 2-azidoethanol (2.00 mmol, 174 mg). Rdto: 520 mg (98%). Yellowish oil. 1H NMR (400 MHz, CDCl3) δ 7.40 (s, 1H), 7.29 (d, J = 8.3 Hz, 2H), 6.80 (d, J = 8.2 Hz, 2H), 4.40 (s, 2H), 4.09 (s, 2H), 3.99 (s, 2H), 3.77 (s, 3H), 3.18 (s, 1H). 13C NMR (101 MHz, CDCl3): δ 159.5, 144.9, 133.9, 125.4, 123.5, 114.8, 61.1, 55.5, 52.9, 30.9. HRMS (ESI +, m / z) calculated
    15 for C12H15N3O2S: 266.0963; Found: 266.0965.
    EXAMPLE 11: 1-Carboxymethyl-4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    Lithium hydroxide monohydrate (2.00 mmol, 84 mg) was added to a solution of 1
    20 methoxycarbonylmethyl-4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (1.00 mmol, 305 mg, example 1) in THF / H2O (1: 1, 8 mL) and the mixture The resulting was stirred at room temperature for one hour. The organic solvent was evaporated, the resulting aqueous mixture was acidified with 1M HCl, and the solution was extracted with EtOAc (2 x 10 mL). The combined organic phases were dried (MgSO4) and the solvent was evaporated under reduced pressure. Rdto: 158 mg (54%). White solid.
    25 P.f .: 163-170 ° C. IR (cm-1): 2923, 2848 (C-H), 1730 (C = O), 1223, 1188 (triazole). 1H NMR (500 MHz, CD3OD): 7.66 (s, 1H), 7.28 (d, J = 8.8 Hz, 2H), 6.84 (d, J = 8.8 Hz, 2H), 5.19 (s, 2H),


    4.07 (s, 2H), 3.76 (s, 3H). 13C NMR (125 MHz, CD3OD): δ 169.7, 161.1, 146.3, 135.6, 126.3, 115.7, 55.8, 51.6, 31.5. HRMS (ESI +, m / z) calculated for C12H14N3O3S: 280.0756; Found: 280.0760.
    EXAMPLE 12: 1-Carboxymethyl-4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of example 11 was followed, starting with 1-methoxycarbonylmethyl-4- [3 (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (1.00 mmol, 305 mg, example 2). Rdto: 274 mg (94%). White solid. Mp .: 115-119 ° C. IR (cm-1): 2999, 2971 (C-H), 1707 (C = O), 1229 (triazole). 1 HOUR
    10 NMR (500 MHz, CD3OD): δ 7.80 (s, 1H), 7.18 (t, J = 8.0 Hz, 1H), 6.96-6.86 (m, 2H), 6, 76 (dd, J = 8.3, 1.8 Hz, 1H), 5.19 (s, 2H), 4.23 (s, 2H), 3.75 (s, 3H). 13C NMR (125 MHz, CD3OD): δ 169.7, 161.4, 146.1, 138.0, 130.9, 125.9, 123.1, 116.1, 113.6, 55.7, 51.7, 29.3. HRMS (ESI +, m / z) calculated for C12H14N3O3S: 280.0756; Found: 280.0753.
    EXAMPLE 13: (S) -1- (1-Carboxy-2-phenylethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of Example 11 was followed, starting from (S) -1- (1-methoxycarbonyl-2-phenylethyl) -4 [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (1.00 mmol , 383 mg, example 3). Rdto: 318 mg (86%). White solid. Mp .: 79-83 ° C. [α] D24 = -23.3 ° (c. 1.12 g / 100 mL, CH2Cl2). IR (cm-1): 2931 (C
    20 H), 1727 (C = O), 1246, 1229 (triazole). 1H NMR (500 MHz, CD3OD): δ 7.77 (s, 1H), 7.20-6.71 (m, 9H), 5.56 (dd, J = 10.8, 4.6 Hz, 1H ), 4.15 (s, 2H), 3.73 (s, 3H), 3.56 (dd, J = 14.3, 4.5 Hz, 1H), 3.40 (dd, J = 14, 3, 10.9 Hz, 1H). 13C NMR (125 MHz, CD3OD): δ 171.1, 161.4, 145.9, 137.9, 137.2, 130.8, 129.9, 128.1, 124.8, 122.9, 115.9, 113.5, 65.8, 55.7, 38.9, 29.0. HRMS (ESI +, m / z) calculated for C20H22N3O3S: 384.1382; Found: 384.1392.


    EXAMPLE 14: (R) -1- (1-Carboxy-2-phenylethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of Example 11 was followed, starting from (R) -1- (1-methoxycarbonyl-2-phenylethyl) -4
    5 [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (1.00 mmol, 383 mg, example 4). Rdto: 280 mg (76%). White solid. Mp .: 78-86 ° C. HRMS (ESI +, m / z) calculated for C20H22N3O3S: 384.1382; Found: 384.1382. [α] D24 = + 16.5 ° (c. 0.98 g / 100 mL, CH2Cl2). The NMR data were identical to those in example 13.
    EXAMPLE 15: (S) -1- (1-Carboxy-3-methylbutyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of Example 11 was followed, starting with (S) -1- (1-methoxycarbonyl-3-methyl-butyl) 4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (1, 00 mmol, 349 mg, example 5). Rdto: 265 mg (76%). White solid. Mp .: 96-105 ° C. [α] D25.6 = + 9.3 ° (c. 1.11 g / 100 mL, CH2Cl2). 1 H NMR (500
    15 MHz, CDCl3): δ 11.88 (s, 1H), 7.54 (s, 1H), 7.12 (t, J = 7.9 Hz, 1H), 6.92-6.77 (m , 2H), 6.70 (dd, J = 8.1, 1.7 Hz, 1H), 5.38 (dd, J = 10.7, 5.0 Hz, 1H), 4.23 (dd, J = 40.8, 14.9 Hz, 2H), 3.70 (s, 3H), 2.03-1.86 (m, 2H), 1.17 (dt, J = 19.8, 6, 5 Hz, 1H), 0.88 (d, J = 6.5 Hz, 3H), 0.82 (d, J = 6.5 Hz, 3H). 13C NMR (125 MHz, CDCl3): δ171.2, 159.8, 144.7, 135.9, 129.8, 122.6, 122.4, 115.6, 112.9, 61.8, 55 , 3, 41.2, 28.5, 24.7, 22.6, 21.1.
    EXAMPLE 16: (R) -1- (1-Carboxy-3-methylbutyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.


    OR
    The procedure of Example 11 was followed, starting from (R) -1- (1-methoxycarbonyl-3-methyl-butyl) 4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (1, 00 mmol, 349 mg, example 6). Rdto: 301 mg (86%). White solid. Mp .: 97-104 ° C. [α] D25.4 = -12.1 ° (c. 0.95 g / 100 mL, CH2Cl2). The NMR data were identical to those in example 15.
    EXAMPLE 17: (S) -1- (1-Carboxy-2-methylpropyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of Example 11 was followed, starting from (S) -1- (1-methoxycarbonyl-2-methyl
    10 propyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (1.00 mmol, 335 mg, example 7). Rdto: 279 mg (87%). White solid. Mp .: 115-121 ° C. [α] D25.6 = + 4.5 ° (c. 1.06 g / 100 mL, CH2Cl2). 1 H NMR (500 MHz, CDCl 3): δ 11.53 (s, 1 H), 7.68 (s, 1 H), 7.12 (t, J = 7.8 Hz, 1 H), 6.87 (d, J = 7.4 Hz, 1H), 6.84 (s, 1H), 6.70 (d, J = 6.8 Hz, 1H), 5.13 (d, J = 7.6 Hz, 1H) , 4.23 (dd, J = 43.5, 14.5 Hz, 2H), 3.71 (s, 3H), 2.44 (d, J = 6.4 Hz, 1H), 0.96 ( d, J = 6.4 Hz, 3H), 0.75 (d, J = 6.4 Hz, 3H). 13C
    15 NMR (125 MHz, CDCl3): δ 170.7, 159.9, 144.6, 136.1, 129.9, 122.9, 122.8, 115.8, 113.0, 69.3, 55.4, 32.2, 28.7, 19.2, 18.3.
    EXAMPLE 18: (R) -1- (1-Carboxy-2-methylpropyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    OR


    The procedure of Example 11 was followed, starting from (R) -1- (1-methoxycarbonyl-2-methyl-propyl) 4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole. (1.00 mmol, 335 mg, example 8). Rdto: 290 mg (90%). White solid. Mp .: 114-123 ° C. [α] D25.6 = -6.7 ° (c. 1.01 g / 100 mL, CH2Cl2). The NMR data were identical to those in example 17.
    EXAMPLE 19: 1- (1-Carboxy-1-methyl ethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    The procedure of example 11 was followed, starting from 4- [3- (methoxy) phenylthiomethyl] -1- (1-methyl-1-methoxycarbonylethyl) -1H-1,2,3-triazole (0.8 mmol, 258 mg, example 9). Rdto: 246 mg (100%).
    10 Yellowish solid. Mp: 109-121 ° C 1H NMR (400 MHz, CD3OD): δ 7.77 (s, 1H), 7.09 (t, J = 8.0 Hz, 1H), 6.82 (d, J = 8 , 0 Hz, 1H), 6.79 (s, 1H), 6.67 (d, J = 8.3 Hz, 1H), 4.12 (s, 2H), 3.65 (s, 3H), 1.78 (s, 6H). 13C NMR (101 MHz, CD3OD) δ 174.2, 161.3, 145.3, 137.9, 130.8, 123.4, 116.5, 113.7, 65.9, 55.7, 29 , 5, 25.9. EXAMPLE 20: 1- [2- (N, N-Dimethylamino) ethyl] -4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole.
    Triethylamine (8.80 mmol, 1.22 mL) and mesyl chloride (4.39 mmol, 0.34 mL) were added successively over a solution of 1- (2-hydroxyethyl) -4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (2.92 mmol, 776 mg, example 10) in anhydrous THF (16 mL) cooled to 0 ° C under a nitrogen atmosphere. The reaction mixture was stirred at room temperature overnight. Next, dimethylamine hydrochloride (8.00 mmol, 652 mg), triethylamine (8.80 mmol, 1.22 mL), NaI (0.13 mmol, 20 mg) and an additional amount of anhydrous THF (8 mL) were added ) and the mixture was stirred at 50 ° C overnight. After completing the reaction, the solvents were evaporated under reduced pressure, the resulting residue was dissolved in EtOAc (20 mL) and washed


    successively with saturated aqueous NaHCO3 solution (30 mL) and brine (10 mL). The organic layer was dried (MgSO4) and evaporated to dryness in vacuo. Rdto: 572 mg (84%). White solid. Mp .: 35-40 ° C. IR (cm-1): 2943 (N-H), 2860, 2771 (C-H), 1242 (triazole). 1H NMR (400 MHz, CDCl3) δ 7.41 (s, 1H), 7.30 (d, J = 7.6 Hz, 2H), 6.81 (d, J = 7.8 Hz, 2H), 4.37 (s, 2H), 4.11 (s, 2H), 3.77 (s, 3H), 2.71 (s, 2H), 2.25 (s, 6H). 13C NMR (101 MHz, CDCl3): 159.2, 144.8, 133.7, 125.5, 122.6, 114.5, 58.6, 55.3, 48.0, 45.3, 30 , 9. HRMS (ESI +, m / z) calculated for C14H21N4OS: 293.1436; Found: 293.1440.
    EXAMPLE 21: 1-Methoxycarbonylmethyl-4- (phenylsulfinylmethyl) -1H-1,2,3-triazole.
    The procedure of Example 1 was followed, starting from phenylthio-2-propine (2.00 mmol, 268 mg) and methyl azidoacetate (2.00 mmol, 230 mg). Rdto: 342 mg (65%). White solid. Mp .: 70-74 ° C. IR (cm – 1): 2953, 2837 (C-H), 1749 (C = O), 1220, 1180 (triazole). 1H NMR (400 MHz, CDCl3): δ 7.49 (s, 1H), 7.34 (d, J = 7.6 Hz, 2H), 7.31-7.22 (m, 2H), 7, 20 (m, 1H), 5.11 (s, 2H), 4.27 (s, 15 2H), 3.78 (s, 3H). 13C NMR (101 MHz, CDCl3): δ 166.5, 144.6, 135.1, 129.1, 128.6, 126.1, 123.5, 52.6, 50.3, 28.3. On the resulting solution of 1- (methoxycarbonylmethyl) -4 (phenylthiomethyl) -1H-1,2,3-triazole (1.00 mmol, 263 mg) in anhydrous chloroform (30 mL) m-chloroperbenzoic acid (1, 00 mmol, 172 mg) at 0 ° C and the mixture was stirred at the same temperature for 30 min. The mixture was evaporated and the residue was purified by chromatography of
    20 column (silica gel; MeOH / CH2Cl2 5:95). Colorless oil Rdto: 161 mg (58%). 1H NMR (500 MHz, CDCl3): δ 7.72 (s, 1H), 7.49 (s, 5H), 5.24-5.10 (dd, J = 5.13 Hz, 2H), 4, 29-4.15 (dd, J = 4.28 Hz, 2H), 3.82 (s, 3H).
    EXAMPLE 22: 1-Carboxymethyl-4- [3- (methoxy) phenylsulfonylmethyl] -1H-1,2,3-triazole.


    On a solution of 1- (carboxymethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole (1.00 mmol, 279 mg, example 12) in a 1: 1 anhydrous mixture of chloroform / acetonitrile (3 mL) m-chloroperbenzoic acid (2.50 mmol, 437 mg) is added at 0 ° C and the mixture is stirred at room temperature overnight. The mixture is evaporated and the residue is purified by column chromatography (silica gel; MeOH / CH2Cl2 5:95). Rdto: 203 mg (65%). White solid. Mp .: 106115 ° C. 1 H NMR (500 MHz, CDCl 3): 7.91 (s, 1 H), 7.50-7.29 (Ar, 4 H), 4.99 (s, 2 H), 4.71 (s, 2 H), 3 , 84 (s, 3H). HRMS (ESI +, m / z) calculated for C12H14N3O5S: 312.0654; Found: 312.0662.
    EXAMPLE 23: 1-Carboxymethyl-4- [3- (chloro) phenylthiomethyl] -1H-1,2,3-triazole.
    A solution of bromoacetic acid (10 mmol, 1.38 g) and sodium azide (40 mmol, 2.60 g) in water (4 mL) was stirred at room temperature overnight. The solution was then neutralized with 3M NaOH and acetonitrile (15 mL), 1- (3-chlorophenylthio) -2-propine (8 mmol, 1.45 g), sodium acetate (30 mmol, 2.46 g) were added successively copper (I) acetate 15 (2 mmol, 242 mg). The resulting mixture was stirred at 45 ° C for 6 h, the solvent was evaporated, acidified with 1M HCl and extracted with EtOAc (3 x 20 mL). The combined organic phases were dried (MgSO4) and the solvent was evaporated under reduced pressure. Rdto: 1.49 g (66%). White solid. Mp .: 146-148 ° C. IR (cm-1): 2928, 2850 (C-H), 1731 (C = O), 1224, 1184 (triazole). 1H NMR (500 MHz, CD3OD): 7.86 (s, 1H), 7.39 (s, 1H), 7.28-7.21 (m, 3H), 5.22 (s, 2H), 4 , 29 (s,
    20 2H). 13C NMR (125 MHz, CD3OD): δ 168.5, 144.1, 137.9, 134.4, 130.0, 128.8, 127.6, 126.3, 124.6 50.4, 27 , 7. HRMS (ESI +, m / z) calculated for C11H10ClN3O2S: 283.0182; Found: 283.0190.
    EXAMPLE 24: Biological tests of in vitro toxicity in human cells.
    25 These tests were carried out in human myotubes LHCN-M2 control after 14 days in differentiation medium. To determine the toxicity of the different compounds analyzed, in particular AHK1 and AHK2 according to the invention, they were added to the culture medium for 24 hours at 37 ° C, at different concentrations (0-2 mM). For comparative purposes, the same was performed


    Assay by adding the reference compound S-107. Cell viability was determined by the Cytotox 96 colorimetric assay (Promega) following the instructions in the manual.
    The results are shown in Figure 1 where it can be seen that the compounds according to the invention do not show acute toxicity in vitro against human myotubes at concentrations between 10 nM and 2 mM. The behavior of the AHK1 and AHK2 compounds after 24 h of incubation contrasts with the 100% cellular toxicity found for the reference compound S-107 at 1 mM concentration under identical conditions. This result demonstrates that the compounds according to the invention have a lower toxicity than S-107, suggesting that AHK compounds may be better candidates for therapeutic treatments of human disorders that involve abnormal calcium homeostasis.
    EXAMPLE 25. In vitro assays to determine intracellular calcium levels in mouse muscle fibers.
    Fibers isolated from the mouse short digital flexor muscle were cultured overnight in the presence or absence of the AHK1 and AHK2 compounds at a concentration of 150 nM. Basal levels of intracellular calcium were evaluated by incubating the fibers with the Fura 2-AM (4 M) ratiometric fluorochrome and pluronic acid (0.02%) for 30 minutes at 37 ° C, in the culture medium. The fibers were visualized with a high resolution digital camera and the intracellular [Ca2 +] was estimated by the excitation ratio 340 nm / 380 nm.
    Figure 2 shows the in vitro effect of the compounds AHK1 and AHK2 on intracellular levels of calcium at rest in the aforementioned mouse muscle fibers. It can be seen how dystrophic fibers that were not treated (MDX) showed a significant increase in calcium levels compared to control fibers (CTRL). Overnight treatment of MDX fibers with AHK1 and AHK2 rescued intracellular calcium levels to control levels demonstrating the ability to reverse the increases in intracellular calcium observed in muscle fibers. In the light of these results, it is postulated that the compounds according to the invention carry out this reversal through a mechanism that involves modulation of RyR channels.
    EXAMPLE 26. In vitro assays on the interaction of RyR1-calstabine 1


    This test was carried out in human myotubes LHCN-M2 control after 9 days in differentiation medium. Said myotubes were pre-treated for 12 hours with the compounds AHK1 and AHK2 at a concentration of 150 nM. After the treatment, the myotubes were subjected to nitro-oxidative stress by peroxitrites by the addition of SIN1 (5 mM) for 30 minutes.
    The colocalization of RyR1-calstabine was analyzed using the in situ proximity ligation technique (PLA in situ) for which the Sigma Duolink II Red Fluorescence Kit was used, and specific antibodies against RyR1 and calstabine 1. This technique allows detect the exact location of two antigens that are less than 40nm from each other. To determine the colocalization of RyR1 with calstabine 1, 3 photographs were quantified for each condition with approximately 9 myotubes per field, using Image J software (http://rsb.info.nih.gov/ij/download.html). In each image, the colocalization area was normalized with the myosin expression area, which was determined by immunofluorescence with a specific fluorescein-conjugated antibody.
    As shown in Figure 3, the compounds AHK1 and AHK2 according to the invention have the capacity to partially recover the decrease in the interaction of RyR1-calstabine1 in healthy human myotube cultures after being subjected to nitro-oxidative stress. The analysis of the RyR1-Calst1 interaction by means of the PLA technique showed that in the presence of SIN1 the dissociation of the RyR1-Calst1 complex occurs and that such dissociation can be partially prevented by pretreatment with the compounds AHK1 and AHK2. In the upper panel of Figure 3 the quantification of the PLA images with ImageJ is shown, while in the lower panel representative images of PLA of each condition are shown, where the points represent the RyR1-Calst1 interaction (calibration bar 50 m). Therefore, the compounds tested according to the invention not only improve the functionality of Duchenne or Becker dystrophic myotubes, but also minimize the degenerative effects of nitro-oxidative stress on healthy human myotubes through an increase in the affinity of the RyR1-Calstabine1 interaction.
    According to these results, the compounds of the invention may be useful as therapeutic agents against diseases caused by the decrease in RyR1-calstabine affinity under conditions of nitro-oxidative stress.
    EXAMPLE 27. Biological assays in vivo in mice


    One month old male mdx dystrophic mice supplied by the Jackson Laboratory (https://www.jax.org/strain/001801) were used. These mice were treated with compound AHK1 or compound AHK2 for 5 weeks, where said compounds were administered in the drinking water at a concentration of 0.25 mg / mL. Weekly, the muscular strength of the front legs was measured using a grip strength meter and the value obtained was normalized by the animal's body weight. To this end, the indications described in the TREAT-NMD Neuromuscular Network protocol (http://www.treat-nmd.eu/downloads/file/sops/dmd/MDX/ DMD_M.2.2.001.pdf) were followed. Dystrophic mice showed a significant decrease in grip strength. However, after 2 weeks of treatment, the strength increased significantly in mdx mice treated with AHK1 or with AHK2 compared to their untreated littermates. After 5 weeks of treatment, muscle strength increased significantly by 20% (Figure 4).
    These data suggest that the compounds according to the invention may be effective in treating patients with muscular dystrophies by improving muscle function or preventing muscle weakness.
    After concluding the 5-week treatment, the anterior tibial muscle was obtained, which was processed for later biochemical and immunohistological analysis to determine the degree of muscle damage. Regeneration derived from cell death was determined in muscle cryosections by quantifying the percentage of central nuclei, using standard immunofluorescence techniques to detect collagen IV and cell nuclei. Figure 5 shows representative cryostat sections of diaphragms of control mice and dystrophic mice marked to see collagen IV and DAPI to see nuclei. The 5-week treatment of mdx mice with AHK1 and AHK2 significantly reduced the percentage of central nuclei, demonstrating the ability of said compounds to reduce histopathological markers of muscular dystrophy after 5 weeks of treatment. These results suggest that the compounds according to the invention are effective in vivo and reach the skeletal muscle.
    For its part, the biochemical analysis of the tibialis anterior muscle was performed by RNA extraction and analysis of the expression pattern in control mice, in mdx mice and in mice subjected to the different treatments. For this, the RT2 Profiler PCR Array specific to human skeletal muscle, myogenesis and myopathy (PAHS-099Z, QIAGEN) was used, using


    cDNA mixtures of at least 3 mice per group. The expression profile of 84 genes involved in pathophysiological mechanisms of skeletal muscle was analyzed. The experiments were performed on the 7300 Real-Time PCR (Applied Biosystems) and the results obtained were analyzed using the QIAGEN online software
    5 (http://www.sabiosciences.com/dataanalysis.php).
    Treatment of mdx mice with the compound AHK1 for 5 weeks partially rescued the gene expression pattern of mdx mice, as shown by comparing the gene expression of WT and mdx mice in Figure 5. The number of genes whose expression was increased in mdx mice with respect to control was 40, which was reduced to 20 in mice
    10 treated with AHK1 (MDX + AHK1). A gene was considered to be overexpressed when its expression increased at least 1.75 times compared to the control. Table I shows the list of genes whose expression is increased in mdx mice and that decreases to control values, which partially decreases or does not change with the treatment of AHK1.
    Table 1
    Unigene Symbol RefSeq Exchange Rate vs. MDX control MDX + AHK1
    Overexpressed genes whose expression is normalized with AHK1
    Akt1 Mm.6645 NM_009652 2.63 1.42 Bcl2 Mm.257460 NM_009741 1.94 1.05 Cast Mm.441995 NM_009817 1.86 1.49 Cav3 Mm.3924 NM_007617 3.28 1.68 Cryab Mm. 178 NM_009964 3 , 08 1.50 Ctnn1 Mm. 291928 NM_007614 1.79 1.27 Dag1 Mm. 491797 NM_010017 2.27 1.23 Des Mm. 6712 NM_010043 2.14 1.53 Dysf Mm. 2020982 NM_021469 2.15 1.28 Foxo3 Mm. 391700 NM_019740 1.76 1.14 Igfbp3 Mm. 29254 NM_008343 1.77 1.34 Igfbp5 Mm.405761 NM_010518 2.52 1.43 Ikbkb Mm.277886 NM_010546 2.72 1.62 Mapk3 Mm. 8385 NM_011952 2, 61 1.69 Myod1 Mm. 1526 NM_010866 4.27 1.28 Nfkb1 Mm. 256765 NM_008689 2.45 1.67 Ppargc1b Mm. 415302 NM_133249 1.77 1.23 Prkaa1 Mm. 207004 NM_001013367 2.33 1.55 Rps6kb1 Mm .446624 NM_028259 1.77 1.25


    Utrn Mm. 418026 NM_011682 2.03 1.35 Overexpressed genes that are partially normalized with AHK1
    Casp3 Mm.34405 NM_009810 10.28 5.27 Igf2 Mm.3862 NM_010514 22.31 9.78 Il1b Mm.222830 NM_008361 17.77 2.82 Il6 Mm.1019 NM_031168 32.72 9.76 Lmna Mm.471227 NM_019390 5 , 29 2.52 Mmp9 Mm.4406 NM_013599 4.51 2.01 Myog Mm.16528 NM_031189 17.36 3.24 Tgfb1 Mm.248380 NM_011577 6.51 3.61 Tnnc1 Mm.439921 NM_009393 25.00 4.53 Tnnt1 Mm.358643 NM_011618 12.09 3.37
    Overexpressed genes whose expression does not vary with AHK1
    Bmp4 Mm. 6813 NM_007554 1.93 2.08Capn2 Mm. 193030 NM_009794 2.90 1.75Igf1 Mm. 2,68521 NM_010512 2.87 2.07Tnf Mm. 1293 NM_013693 15.01 10.27Prkag3 Mm. 453463 NM_153744 2.52 2.68Rhoa Mm. 318359 NM_016802 2.30 1.80Pax3 Mm. 1371 NM_008781 2.16 2.48Pax7 Mm. 21760 NM_011039 3.82 2.55Musk Mm. 16148 NM_010944 3.51 2.00Myf5 Mm. 4984 NM_008656 3.40 2.13
    Specifically, the results show that the compounds according to the invention show capacity to reduce by half the number of overexpressed genes in mdx dystrophic muscle. Genes likely to be modulated by AHK compounds include, but not
    5 are limited to, Akt1, Bcl2, Casp3, Cast, Cav3, Cryab, Ctnnb1, Dag1, Des, Dysf, Foxo3, Igfbp3, Igfbp5, Ikbkb, Mapk3, Myod1, Nfkb1, Ppargc1b, Prkaa1, Rf6, Castr3, Rf6 Castr , Il1b, Il6, Lmna, Mmp9, Myog, Tgfb1, Tnnc1 and Tnnt1. Within this list, Akt1, Il1b, Mapk3, Mmp9 and Utr are genes related to skeletal muscle loss or atrophy.

    权利要求:
    Claims (20)
    [1]
    1. A compound of Formula (I):
    (OR)NN
    n
    N R2Y
    S R1
    (X)
    m
    5 R3
    (I)
    where
    R1 is a birradical C1-4 alkylene optionally substituted by one or more substituents independently selected from the group consisting of C1-4 alkyl, C6-10 aryl, F, Cl, CN and
     10 NO2;
    R2 is a biradical C1-6 alkylene, in which 1, 2 or 3 groups -CH2- can be optionally replaced by groups selected from –O– and –S–; and wherein the biradical C1-C6 alkylene may be optionally substituted with one or more groups independently selected from the group consisting of C1-4 alkyl, allyl, propargyl, hydroxymethyl, 1
    15 hydroxyethyl, 2-hydroxyethyl, 3-aminopropyl, 4-aminobutyl, 3-guanidylpropyl, 3-indolylmethyl, C6-10 aryl, benzyl, 4-hydroxybenzyl, C6-10 heteroaryl, F, Cl, OH, O (C1- alkyl 4), CN, NO2, CO (C1-4 alkyl), CO2 (C1-4 alkyl), -CONH (C1-4 alkyl) and -CON (C1-4 alkyl) 2;
    R3 is selected from the group consisting of H, C1-4 alkyl, C6-10 aryl, F, Cl, Br and I;
    m is selected from 0, 1, 2 and 3, 4;
    20 n is selected from 0, 1 and 2;
    X is independently selected from the group consisting of OH, O (C1-4 alkyl), O (C610 aryl), OCF3, S (C1-4 alkyl), S (C6-10 aryl), C1-6 alkyl, CF3, NHC (O) (C1-4 alkyl) and halogen; or two X groups may represent a birradical methylenedioxy, ethylenedioxy or propylenedioxy; and
    Y is selected from the group consisting of -OH, -CO2H, -CO2 (C1-4 alkyl), -CO2 (allyl),
    CO2 (benzyl), -SO3H, -NH2, -NH (C1-4 alkyl), -N (C1-4 alkyl) 2, N (C1-4 alkyl) 3 and N (heterocyclyl or heteroaryl), wherein said heterocyclyl or heteroaryl is found

    optionally substituted by a C1-4 alkyl group and where the N atom is part of the heterocyclyl or heteroaryl,
    or a stereoisomer, pharmaceutically acceptable salt, solvate or complex thereof, or an isotopically labeled derivative or a prodrug thereof.
    Compound according to claim 1, wherein R1 is -CH2-.
    [3]
    3. Compound according to any of the preceding claims, wherein R2 is a birradical C1-4 alkylene optionally substituted with one or two substituents independently selected from the group consisting of methyl, ethyl, propyl, isopropyl, allyl, propargyl, butyl, isobutyl, sec-butyl, tert-butyl, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 3
    10 aminopropyl, 4-aminobutyl, 3-guanidylpropyl, 3-indolylmethyl, phenyl, 1-naphthyl, 2-naphthyl, benzyl, 4-hydroxybenzyl, C6-10 heteroaryl.
    [4]
    4. Compound according to claim 3, wherein R2 is a birradical -CH2-or -CH2-CH2-, optionally substituted with one or two substituents independently selected from the group consisting of methyl, isopropyl, isobutyl and benzyl.
    5. A compound according to claim 4, wherein R2 is a birradical -CH2-optionally substituted with two methyl substituents or with a substituent selected from the group consisting of isopropyl, isobutyl and benzyl, or wherein R2 is a birradical -CH2- CH2-.
    [6]
    6.  Compound according to claim 5, wherein R2 is a birradical -CH2-or -CH2-CH2-.
    [7]
    7.  Compound according to any of the preceding claims, wherein R3 is H.
    A compound according to any one of the preceding claims, wherein m 1.
    [9]
    9.  Compound according to any of the preceding claims, wherein n is 0.
    [10]
    10.  Compound according to any of the preceding claims, wherein X is O (C14 alkyl) or halogen.
    [11]
    11. A compound according to claim 10, wherein X is a methoxy group in the meta position or para or chlorine.
    [12]
    12. Compound according to any of the preceding claims, wherein Y is selected from the group consisting of CO2H, CO2Me, NH2, -NHMe, -NMe2, -NMe3, -NHEt, -NEt2, -NEt3,

    or a pharmaceutically acceptable salt of said groups.
    [13]
    13. Compound according to claim 12, wherein Y is selected from the group consisting of CO2H, CO2Me, NH2, -NHMe, -NMe2, -NMe3, -NHEt, -NEt2, -NEt3, pyrrolidin-1-yl, piperidin-1yl , morpholin-4-yl, 4-piperazin-1-yl, 4-methyl-piperazin-1-yl and pyridin-1-yl or a salt
    5 pharmaceutically acceptable from said groups.
    [14]
    14. Compound according to claim 13, wherein Y is selected from the group consisting of CO2H and -NMe2.
    [15]
    fifteen. Compound of formula (I) according to claim 1, selected from the group consisting of: 1-carboxymethyl-4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole,
    10 1- [2- (N, N-dimethylamino) ethyl] -4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, 1-carboxymethyl-4- [4- (methoxy) phenylthiomethyl ] -1H-1,2,3-triazole, (S) -1- (1-carboxy-2-phenylethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, ( R) -1- (1-carboxy-2-phenylethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, (S) -1- (1-carboxy-3-methylbutyl ) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole,
    15 (R) -1- (1-carboxy-3-methylbutyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, (S) -1- (1-carboxy-2 -methylpropyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, (R) -1- (1-carboxy-2-methylpropyl) -4- [3- (methoxy) phenylthiomethyl ] -1H-1,2,3-triazole, 1- (1-carboxy-1-methyl ethyl) -4- [3- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole, 1- (2- hydroxyethyl) -4- [4- (methoxy) phenylthiomethyl] -1H-1,2,3-triazole,
    1-Methoxycarbonylmethyl-4- (phenylsulfinylmethyl) -1H-1,2,3-triazole, and 1-carboxymethyl-4- [3- (methoxy) phenylsulfonylmethyl] -1H-1,2,3-triazole, 1-carboxymethyl -4- [3- (chloro) phenylthiomethyl] -1H-1,2,3-triazole,
    or a salt thereof.
    [16]
    16. A pharmaceutical composition comprising a compound of Formula (I) as described
    Defined in any of claims 1-15, and one or more pharmaceutically acceptable excipients or vehicles.

    [17]
    17. Pharmaceutical composition according to claim 16 for oral or parenteral administration.
    [18]
    18. Use of a compound of formula (I) as defined in any one of claims 1 to 15 to prepare a medicament.
    19. Use of a compound of formula (I) as defined in any one of claims 1 to 15 to prepare a medicament for the treatment and / or prevention of skeletal muscle disorders and diseases, cardiac disorders and diseases and disorders and diseases of the nervous system.
    [20]
    20. Use according to claim 19, wherein skeletal muscle disorders and diseases are
    10 select between muscular dystrophies, congenital myopathies, metabolic myopathies, muscular atrophy and sarcopenia.
    [21]
    21. Use according to claim 20, wherein the skeletal muscle disorder or disease is Duchenne muscular dystrophy or Becker muscular dystrophy.
    [22]
    22 Use according to claim 19, wherein the cardiac disorders and diseases are selected from heart failure, cardiac ischemia, cardiac arrhythmias and cardiomyopathies.
    [23]
    23. Use according to claim 19, wherein disorders and diseases of the nervous system are selected from brain accident, Alzheimer's disease, frontotemporal dementia and cognitive impairment.
    [24]
    24. A method for the synthesis of a compound of Formula (I) as defined in any one of claims 1-15, comprising:
    a) reacting an alkyne of formula (II) with an azide of Formula (III),
    n
    R1 R3
    N3 R2Y
    (II) (III)
    optionally in the presence of a copper catalyst and, optionally, in the presence of a base to give a compound of formula (I),
    where:

    the groups R1, R2, R3, m, n and X in the compounds of formulas (II) - (III) are as defined in any of claims 1-15, and
    Y is a group as defined in any one of claims 1-15, optionally protected with a carboxyl protecting group, a hydroxyl protecting group or an amino protecting group;
    b) when n is 0 in the compound of formula (I) obtained in step a), optionally treating said compound of formula (I) with an oxidizing agent to give a compound of formula (I) wherein n is 1 or 2 , R1, R2, R3, and X are as defined in any of claims 1-15, and Y is a group as defined in any of
    10 claims 1-15, optionally protected with a carboxyl protecting group, a hydroxyl protecting group or an amino protecting group; Y
    c) when the compound of formula (I) obtained in step a) or b) has a Y group protected with a protective group, remove said protective group to give a compound of formula (I) wherein R1, R2, R3, m , n, X and Y are as defined in any
    15 of claims 1-15.

    FIGURES
    FIGURE 1

    FIGURE 2

    FIGURE 3

    FIGURE 4

    FIGURE 5

    FIGURE 6
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    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    WO2013142346A1|2012-03-23|2013-09-26|The Regents Of The University Of California|Premature-termination-codons readthrough compounds|
    WO2014205414A1|2013-06-21|2014-12-24|The Regents Of The University Of California|Expanded therapeutic potential in nitroheteroaryl antimicrobials|
    EP2857388A1|2013-10-01|2015-04-08|Grünenthal GmbH|Azoles containing sulfone|
    WO2016057656A1|2014-10-08|2016-04-14|Mitobridge, Inc.|Ppar-delta agonists for use for treating mitochondrial, vascular, muscular, and demyelinating diseases|
    JP2703408B2|1990-12-28|1998-01-26|麒麟麦酒株式会社|1,4-benzothiazepine derivatives|
    US8022058B2|2000-05-10|2011-09-20|The Trustees Of Columbia University In The City Of New York|Agents for preventing and treating disorders involving modulation of the RyR receptors|
    US20040229781A1|2000-05-10|2004-11-18|Marks Andrew Robert|Compounds and methods for treating and preventing exercise-induced cardiac arrhythmias|
    EP1751120A4|2004-05-25|2010-05-05|Metabolex Inc|Substituted triazoles as modulators of ppar and methods of their preparation|
    US7414134B2|2004-10-12|2008-08-19|Amgen Inc.|B1 bradykinin receptor antagonists|
    US7515666B2|2005-07-29|2009-04-07|International Business Machines Corporation|Method for dynamically changing the frequency of clock signals|
    US7704990B2|2005-08-25|2010-04-27|The Trustees Of Columbia University In The City Of New York|Agents for preventing and treating disorders involving modulation of the RyR receptors|
    WO2008060332A2|2006-06-02|2008-05-22|The Trustees Of Columbia University In The City Of New York|Methods for treating or reducing muscle fatigue|
    US8703804B2|2006-03-26|2014-04-22|Uti Limited Partnership|Ryanodine receptor inhibitors and methods relating thereto|
    WO2012019071A1|2010-08-06|2012-02-09|The Trustees Of Columbia University In The City Of New York|Methods of preventing and treating sarcopenia|
    WO2012037105A1|2010-09-14|2012-03-22|The Trustees Of Columbia University In The City Of New York|Methods of treating, ameliorating or preventing stress-induced neuronal disorders and diseases|
    EP2708535A1|2012-05-11|2014-03-19|Les Laboratoires Servier|Agents for treating disorders involving modulation of ryanodine receptors|TW201808345A|2016-05-24|2018-03-16|美商安美基公司|Pegylated carfilzomib compounds|
    WO2021162054A1|2020-02-10|2021-08-19|学校法人順天堂|Type 2 ryanodine receptor activity inhibitor|
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