TW200815381A - Methods for treating or reducing muscle fatigue - Google Patents

Abstract

The invention provides methods for treating muscle fatigue with compounds of the invention. The invention relates to compositions and methods for treating, preventing or reducing muscle fatigue by administering compounds that stabilize ryanodine receptors (RyR), which regulate calcium channel functioning in cells. The invention provides methods to treat muscle fatigue in a subject suffering from a wide variety of acute and chronic pathologies, neurological or genetic diseases or conditions, including but not limited to cardiac disease, HIV infection and AIDS, muscular dystrophy, cancer, malnutrition, exercise-induced muscle fatigue, age-associated muscle fatigue, renal disease and renal failure. The invention provides methods for treating a subject suffering from muscle fatigue as a result of sustained, prolonged and/or strenuous exercise, or chronic stress.

Description

200815381 IX. Description of the invention: [Technical field to which the invention pertains] The field of the invention belongs to skeletal muscle fatigue, muscle symptoms and abnormalities, and the above-described treatment methods. [Prior Art] Sarcoplasmic reticuium (SR) is a specialized storage site for intracellular calcium ions (Ca2+), among other functions. It can open and close the channel of the sarcoplasmic reticulum (ryan〇dine reeeptQi>,

RyR) ' regulates the release of Ca2+ from the sarcoplasmic reticulum into the cytoplasm of cells. The myometrial network releases Ca2+ to the cytoplasm to increase the cytoplasmic Ca2+ concentration. The open probability (Po) of the RyR receptor means the possibility that the RyR channel opens at any given time and is therefore capable of releasing Ca2+ from the sarcoplasmic reticulum to the cytoplasm. There are three types of Arino base receptors, all of which are Ca2+ channels: RyRl, RyR2 and RyR3. RyRl is mainly found in skeletal muscle and other tissues; RyR2 is mainly found in the heart and other tissues; and RyR3 is found in the brain and other tissues. The RyR channel is composed of four RyR polypeptides combined with four FK506 binding proteins (FKBP), which are called FKBP12 (calstabinl) and FKBP12.6 (calstabin2). Calstabinl binds to RyRl; calstabin2 binds to RyR2; and calstabinl binds to RyR3. The FKBP protein (calstabinl and cals tab in2) binds to the RyR channel (one molecule per RyR subunit) to stabilize the channel function of RyR and facilitate coupled gating between adjacent RyR channels; thus avoiding channel closure Abnormal activation in the state. 5 200815381 Protein kinase A (PKA) binds to the cytoplasmic surface of the RyR receptor. Protein stimulating of the RyR receptor A phosphorylation causes partial calstabin to be isolated from RyR. The separation of Calstabin from RyR causes an increase in the RyR opening rate, thus increasing the release of sarcoplasmic reticulum into the cytoplasmic Ca2+ skeletal muscle cells and cardiac cells. The release of Ca2+ from the sarcoplasmic reticulum is an important physiological mechanism for controlling muscle function, because of increased cytoplasm. The Ca2+ concentration can cause muscle contraction. Excitation-contraction (EC) coupling includes electrical depolarization of the cell membrane in a transverse tubule (T-tubule), which activates a voltage-gated L-type Ca2+ channel ( L-type Ca2+ channels, LTCCs). LTCCs trigger sarcoplasmic reticulum to release Ca2+ through physical interaction with RyRl. The increase in intracellular Ca2+ concentration results in the interaction of actin-myosin and muscle contraction. To alleviate muscle, intracellular Ca2+ is pumped back to the sarcoplasmic reticulum via sarcoplasmic reticulum Ca2 and ATPase pumps (SERCAs), which are regulated by phospholamban (PLB), which depends on the type of phosphoprotein Muscle fiber type. Fatigue is a process in which skeletal muscle becomes weaker, either as a result of repeated or intense use (eg, exercise), or as a result of discomfort, abnormality, or disease. Fatigue can cause work failure and is a clear symptom of many medical conditions, including heart failure, renal failure, cancer, and many muscular dystrophys. Over the past decade, it has become clear that two typical muscle fatigue explanations (called 6 200815381 lactic acid accumulation and intracellular acidosis) do not cause fatigue. In fact, these two are protective mechanisms for avoiding fatigue in highly intense exercise (Allen and Westerblad 2004; Pedersen, Nielsen et al. 2004). Muscle contraction depends on the effective coupling of the following processes: electrical stimulation of the muscle surface; release of Ca2+ via the sino base receptor (sarcoplasmic reticulum Ca2+ release channel); to produce the myosin crossover bridge (cross bridge). Thus, it is apparent that the defect causing the transient Ca2+ amplitude reduction in the excitatory contraction coupling, among other effects, causes the contraction and strength to be impaired through the formation of the inefficient cross-linking of the actin. Eberstein and Sandow propose a possible factor that inhibits Ca2+ release as a fatigue process (Eberstein and San do w 1 963). Many muscle preparations have described a reduction in Ca2+ amplitude caused by fatigue stimulation (Allen, Lee et al. 1989; Westerblad and Allen 1991; Allen and Westerblad 2001) and found time course of self-fatigue recovery (time course) ) is similar to the time course of prolonged Ca2+ release (Westerblad, Bruton et al. 2000). Sarcoplasmic reticulum Ca2+ leakage was demonstrated in the myofibrils and muscle wasting patterns after intense exercise (Wang, Weisleder et al. 2005), and this may be due to defects in the skeletal muscle aroma base receptor (RyRl). caused. Long-term activation of the sympathetic nervous system (SNS) in cardiac exhaustion causes skeletal muscle fatigue in the body, because this is due to the lack of phosphodiesterase PDE4D3 on the RyRl complex; protein kinase A versus RyRl The serine 2844 (Serine 2844) hyperphosphorylation; the RyRl complex lacks calstabinl; and the functional increase of 7200815381 (gain-of-function) channel defects (Reiken, Lacampagne et al. 20 03). RyR1 dysfunction in skeletal muscle results in localized subcellular Ca2+ release and reduced global calcium ion transient currents (Ward, Reiken et al. 2 003) 〇JTV5 19,4-[3-(4-Benzyl) Isopiperidin-1-yl)propanyl]-7-methoxy-2,3,4,5-tetrahydro-1,4-benzothiazepine hydrochloride (4_[3-(4) -benzylpiperidin-l-yl)propionyl]-7-methoxy_2,3,4, 5-tetrahydro-1,4-benzothiazepine monohydrochloride), a 1,4-benzothiazepine, already It is shown to be a modulator of the RyR calcium channel, which can be applied to the murine model of heart failure to improve skeletal muscle function, and can be separated by ex vivo separation (five weeks after left coronary artery ligation) verification. The beneficial effect of JTV5 19 on muscle fatigue is not only due to improved cardiac function, but a beneficial effect can still be seen in the administration of the agent in the absence of calstabin2 mice, which do not receive any cardiac benefit from treatment. Therefore, it has been hypothesized that JTV5 19 can directly affect muscle function (Wehrens, Lehnart et al. 2005). In long-term exercise, the phosphodiesterase PDE4D3 is absent on the RyRl complex; protein kinase A is hyperphosphorylated against RyR1's serine 2844; the RyRl complex lacks the same changes in the RyRl macromolecular complex such as calstabinl, and mice There is a relationship between time dependence and activity dependence between the repeated strong movements in the model. After prolonged exercise, the biochemical changes in the regulation and function of these RyRl macromolecular complexes are stable and slow (several days to several weeks). Therefore, it has been suggested that Ca2+ leakage of RyRl limits the peak of muscle performance and causes muscle damage during prolonged and intense exercise. 8 200815381 The contraction of striated muscle begins when a small tube (called sarcoplasmic reticulum (SR)) in muscle cells releases maternal ions (Ca2+). Excitatory contraction (EC) coupling requires a calcium ion release channel (called the RyR) on the muscle network. The second type of the RyR receptor (RyR2) was found in the heart, and the first type of RyR1 receptor (RyR 1) was found in skeletal muscle. The RyRl receptor is a four-molecular polymer consisting of four 565,000 daltons (1&11〇11) 1^111 polypeptide and four 12,000 dalton FK506 binding proteins (FKBP12). FKBP12 is Regulatory subunits that stabilize RyR channel function (Brillantes et al., 1994) and help coupler gating in adjacent RyR channels (Marx et al., 1998); the latter (RyRl) are localized in specific sarcoplasmic reticulum regions (release) The dense array of intracellular Ca2+ stores promotes muscle contraction. In addition to FKBP12, the RyRl macromolecular complex also includes catalytic and regulatory subunits of protein kinase A and luciferase PP1 (Marx et al. , 2001) 〇 Each RyRl subunit binds to a FKBP12 molecule. The separation of FKBP12 significantly alters the biophysical properties of the channel, causing the appearance of a subconductance state and increasing the channel opening rate (P.) (Brillantes et al. 5 1994; Gaburjakova et al., 2001). The separation of FKBP12 from the RyRl channel inhibits coupled gating, resulting in random gate gating rather than ensemble gating (Marx et al., 1 998). Coupling of RyR channel arrays Control is important for the activation of contractile coupling that effectively regulates muscle contraction (Marx et al., 1 998). The FKBP is a cis-trans-plyidyl-prolyl isomerase, which is widely Performance and role in a variety of cellular functions (Marks, 1996). FKBP12 binds tightly and regulates 9 200815381 Controlling the function of the following proteins: skeletal muscle Ca2+ release channel (RyRl) (Brillantes et al., 1994; Jayaraman et al.? 1992) Myocardial Ca2+ release channel (RyR2) (Kaftan et al., 1996); and related intracellular Ca2+ release channels, known as type 1 1,4,5-trisphosphate receptors ( Inositol 1,4,5-triphosphate receptor, IP 3 R 1) (Cameron et al., 1997), and type 1 transforming growth factor p (TGFP) receptor (TpRI) (Chen et Al·, 1997).

/ \ US co-pending application No. 1 〇 / 794, 21 8 discloses a method for treating skeletal muscle function defects in heart failure by administering an agent that inhibits Fkb 1 2 binding protein from the RyRl receptor, This is incorporated herein by reference in its entirety. U.S. Patent Application Serial No. 11/212, No. 3, and U.S. Application Serial No. 11/212,413, disclose the use of the novel benzothiazepines & m and methods for preventing abnormalities and diseases associated with RyR dimers, wherein the abnormalities are disease. Including skeletal muscle dysplasia and disease (such as skeletal muscle fatigue, exercise-induced skeletal muscle fatigue, muscular dystrophy, sputum field fire bladder abnormalities and incontinence), which is hereby incorporated by reference in its entirety. In this article. There is an urgent need to find novel agents that can effectively treat sputum + or prevent muscle fatigue, while muscle fatigue can be caused by stress or fish-to-fish or by the RyR receptor (regulating intracellular calcium channel function). Caused by the disease, including heart disease or abnormality, Dyracic muscle function deficiency, HIV Infection, malnutrition AIDS, muscular dystrophy, cancer, camp Exercise-induced muscle fatigue, age-related 10 200815381 Muscle fatigue, kidney disease, kidney failure. SUMMARY OF THE INVENTION In one aspect, the invention provides a method of treating, preventing or ameliorating a muscle condition, disease or disorder, the method comprising administering to the individual a therapeutically effective amount of a compound of formula I. In certain aspects, the invention provides a method of treating, preventing or ameliorating muscle fatigue in an individual comprising administering to the individual a therapeutically effective amount of a compound having the structure of Formula I;

Where η is 0, 1, or 2; q is 0, 1, 2, 3, or 4; each R system is selected from the group consisting of: Η, halogen, -OH, -ΝΗ2, -NO , -CN, -CF3, -OCF3, -N3, -S03H, -S( = 0)2 alkyl, -S( = 0)alkyl, -0S( = 0)2CF3, fluorenyl, -Ο-醯Base, alkyl, alkoxy, alkylamino, alkylarylamino, thiol, cycloalkyl, aryl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, Alkenyl, alkynyl, (hetero-)aryl, 11 200815381 (hetero-) arylthio and (hetero-)arylamino; wherein thiol, -fluorenyl, alkyl, alkoxy, a thiol group, an alkylthio group, a cycloalkyl group, an alkylaryl group, an aryl group, a aryl group, a hetero-alkyl group, a sulfhydryl group, a fast group, a (hetero-) aryl group and a (hetero-) arylamine group; Among the groups consisting of: jj, aryl, aryl, aryl, cycloalkyl, heteroaryl, may be substituted or unsubstituted, each alkyl, alkenyl, _cycloalkyl, heteroaryl and Heterocyclic group; R2 is a group consisting of %, -C(=0)R5, -C(=S)R6, -S〇2R7-(CH2)m-R10, alkyl, aryl, alkane Fang , cycloalkylalkyl and heterocyclyl · unsubstituted individual alkyl, aryl, alkaryl, heterocycloalkylalkyl and heterocyclic; R3 is a group consisting of the following Selected: -C( = 0)NHY, fluorenyl, -0-fluorenyl, alkyl, aryl, cycloalkyl, heteroaryl and heterocyclic; in which each thiol, alkyl, alkenyl , aryl, arylheteroaryl and heterocyclic; wherein Y is selected from the group consisting of hydrazine, alkyl, aryl, alkaryl, cycloalkylheterocyclyl, and which may or may not be substituted Alkaryl, cycloalkyl, heteroaryl and heterocyclic; or unsubstituted individual, alkylarylamine heteroaryl, heterocycle, (hetero-)arylthioketone, alkyl, heterocyclic ; its base, alkaryl, selected: -P (=0) R8R9 n 1 group, cycloalkyl, which may be substituted or substituted, cycloalkyl, Η, -C〇2Y, aryl, aryl, burn Substituted or unsubstituted, cycloalkyl, constituting group, heteroaryl and aryl, alkane 12 200815381 R4 is selected from the group consisting of hydrazine, alkyl, alkenyl, aromatic Base, aryl group, naphthenic , Aryl, heteroaryl and heterocyclyl; wherein each preferably a substituted or unsubstituted alkyl group, alkenyl group, aryl group, alkaryl group, cycloalkyl, heteroaryl and heterocyclyl;

Rs is selected from the group consisting of -NUu, -(CH2)ZNR15R16, _NHNR"Ri6, ΝΗ〇Η, 〇Ri5,

-C( = 0)NHNR15Ri6> -C〇2r15. -C( = 0)NRi5Ri6' -CH2X . Mercapto, alkyl, alkenyl, aryl, alkaryl, cycloalkyl, cycloalkylalkyl, a heteroaryl group, a heterocyclic group and a heterocycloalkyl group; which may be substituted or unsubstituted, each fluorenyl group, alkyl group, alkenyl group, aryl group, alkylaryl group, cycloalkyl group, cycloalkylalkyl group, heteroaryl group, a heterocyclic group and a heterocycloalkyl group, wherein Z is 1, 2, 3, 4, 5 or 6; R6 is selected from the group consisting of -OR! 5, -NHNR15R16, -NHOH, -NR15R16 , -CH2X, fluorenyl, dilute, alkyl, aryl, alkaryl, cycloalkyl, cycloalkylalkyl, heteroaryl, heterocyclyl and heterocycloalkyl; wherein each may be substituted or unsubstituted Mercapto, alkenyl, alkyl, aryl, alkaryl, cycloalkyl, cycloalkylalkyl, heteroaryl, heterocyclyl and heterocycloalkyl; R7 is selected from the group consisting of : a 0Rl5, -NR15R16, -NHNR15R16, -NHOH, -CH2X, alkyl, alkenyl, alkynyl, aryl, alkaryl, cycloalkyl, cycloalkylalkyl, heteroaryl, heterocyclic and Heterocycloalkyl; which may or may not be substituted for each alkyl, alkenyl, Alkynyl, aryl, alkaryl, cycloalkyl, cycloalkylalkyl, heteroaryl, heterocyclyl and heterocycloalkyl; 13 200815381

The Rs and R9 systems are respectively selected from the group consisting of OH, decyl, alkenyl, alkoxy, alkyl, alkylamino, aryl, aryl, cycloalkyl, cycloalkylane a heteroaryl group, a heterocyclic group and a heterocycloalkyl group; wherein each of a decyl group, an alkenyl group, an alkoxy group, an alkyl group, an alkylamino group, an aryl group, an alkylaryl group, a cycloalkyl group, a cycloalkylalkyl group, a heteroaryl group, a heterocyclic group and a heterocycloalkyl group;

Rio is selected from the group consisting of -NRi5R16, OH, -S02Rn, -NHSO2R11, C(=0)(R12), NHC = 0(R12), -OC = O(R12) and -P ( = 〇) R13 R14; R11, R12, R13 and R14 are respectively selected from the group consisting of hydrazine, OH, NH2, -NHNH2, -NHOH, fluorenyl, fluorenyl, alkoxy, alkyl An alkylamino group, an aryl group, an alkylaryl group, a cycloalkyl group, a cycloalkylalkyl group, a heteroaryl group, a heterocyclic group and a heterocycloalkyl group; wherein the fluorenyl group, the alkenyl group, the alkoxy group may be substituted or unsubstituted a group, an alkyl group, an alkylamino group, an aryl group, an alkylaryl group, a cycloalkyl group, a cycloalkylalkyl group, a heteroaryl group, a heterocyclic group and a heterocycloalkyl group; the X system is selected from the group consisting of the following: : halogen, -CN, -CO2R15, -C( = 0)NRl5Rl6, -NR15R16, -〇Ri5, -S〇2r7 and -P( = 〇)R8R9;

Ri5 and Ri6 are respectively selected from the group consisting of hydrazine, fluorenyl, alkenyl, alkoxy, OH, hydrazine, alkyl, alkylamino, aryl, alkaryl, cycloalkyl, a cycloalkylalkyl group, a heteroaryl group, a heterocyclic group and a heterocycloalkyl group; wherein each of the fluorenyl group, the alkenyl group, the alkyl group, the alkyl group, the alkyl group, the aryl group, the alkylene group may be substituted or unsubstituted. a group, a cycloalkyl group, a cycloalkylalkyl group, a heteroaryl group, a heterocyclic group and a heterocycloalkyl group; and, optionally, R15 and R16 may form a heterocyclic ring together with the nitrogen to which they are bonded, and the heterocyclic ring may Containing a substituent; the gas in the benzothiazepine ring may be optionally a quaternary nitrogen; and its enantiomers, diastereomers, tautomers (tautomers), pharmaceutically acceptable salts, hydrates, solvates, complexes (c 〇 mp 1 e X es) and prodrugs. In certain embodiments, the compound is not SI, S2, S3, S4, S5, S6 > S7, S9, S11, S12, S13, S14, S19, S20, S22, S23, S24, S25, S26, S27, S36, S37, S38, S40, S43, S44, S45, S46 'S47, S48, S49 'S50, S51, S52, S53, S54, S55, S56, S57, S58 'S59, S60, S61, S62, S63, S64 ' S66 ' S67 , S68 ' S69 , S70 ' S71 , S72 , S73 , S74 ' S75 , S76 , S77 , S78 , S79 , S80 , S81 , S82 , S83 , S84 , S85 , S86 , S87 , S88 , S89 , S90, S91, S92, S93, S94, S95, S96, S97, S98, S99 or S100. In another embodiment, the compound is not S4, S7, S20, S24, S25, S26, S27 or S36. In one embodiment, the compound is not JTV-519. In certain embodiments, the methods treat symptoms. In other embodiments, the methods treat the cause and 7 or symptoms. 15 200815381 In certain embodiments 'the compound is described by a chemical formula selected from the group consisting of: Formula Ia, 〗 〖b, ic, 卜d, he, if, Ig, Ih, Ii, Ij , Ik, 1-1, ^, ϊ_η, 1〇 or Ι-Ρ. In other embodiments, the compound is described by a chemical formula selected from the group consisting of: Formula Ia, Ib, Ie, u,, h, Μ, η, 1-0, and Ι-Ρ . In other embodiments, the compound is described by a chemical formula selected from the group consisting of: Formula I-n, Lo and. In other embodiments 'the compound is selected from the group consisting of: S 1 , s 2, S 3, S4, S5, S6, S7, S9, S11, S12, S13, S14, S19, S20, S22 , S23, S24, S25, S26, S27, S36, S37, S38, S40, S43, S44, S45, S46, S4 7, S48, ! 549, S50, S51, S52, S53, S54, S55, S56, S57, S58, S59, S60 'S61, S62, S63, S64, S66, S67, S68, S69, S70, S71, S72, S73, S74, S75, S76, S77, S78, S79, S80, S81, S82, S83, S84, S85, S86, S87 'S88, S89, S90, S9, S92, S93, S94, S95, S96, S97, S98, S99, S100, S101, S102, S103, S104, S107, S10 8, S109, S110, Sm, S112, S113, S114, SI 15, SI 16 > SI 17 > SI 18 > SI 19 ' S120, S121, SI 22 with S 1 23 . In other embodiments, the compound is selected from the group consisting of: S47, S48, S50, S51, S59, S64, S74, S75, S77, S85, S101, S102' S103' S107' S109, S110 'Sill, S117 and S 1 2 1. In other embodiments, the compound is selected from the group consisting of S68, S101, S102, S103, S107, S109, S110, sill, S11 7 and S 1 2 1 . In one embodiment, the compound is s 1 0 7 . 16 200815381 In other embodiments of the method, muscle fatigue is caused by a skeletal muscle disease or abnormality. In other embodiments, skeletal muscle disease or abnormalities are associated with RyRl^: dysfunction. In certain embodiments, the methods treat muscle fatigue caused by myopathy. In other embodiments, the methods treat muscle fatigue caused by muscular dystrophy. In other embodiments, these methods treat muscle fatigue caused by central core disease. In other embodiments, the methods treat malignant hyperthermia. In other embodiments, muscle fatigue is caused by muscle fatigue or age-related muscle fatigue. In other embodiments, exercise-induced muscle fatigue is caused by prolonged exercise or high-intensity exercise. In other embodiments, the individual has neuropathy, neurological disease or abnormality, seizure condition, genetic disease or abnormality, heart disease or abnormality, infectious disease, HIV infection, AIDS, myopathy, Muscular atrophy, cancer, malnutrition, kidney disease or kidney failure. In certain embodiments, the heart disease or abnormality is selected from the group consisting of irregular heartbeat disease, exercise-induced irregular heartbeat, congestive heart failure, chronic obstructive pulmonary disease, hypertension Or any combination of the above. In other embodiments, the irregular heartbeat disorder is selected from the group consisting of: atrial or ventricular arrhythmia; atrial fibrillation or ventricular fibrillation; atrial tachyarrhythmias Or ventricular tachyarrhythmias; atrial tachycardias or heart to frequency 17 200815381 ventricular (tachycardias); catecholaminergic polymorphic ventricular tachycardia (CP VT) and the above-mentioned movement The resulting variant. In other η applications, 'muscle lesions are congenital myopathy. In other embodiments, the myopathy is selected from the group consisting of: muscular dystrophy, central axis disease, myopathy with cores and rods, mitochondrial myopathy (mitochondrial myopathy), endocrine myopathy, glycogen storage diseases, myoglobinurias, dermatomyositis, myositis ossificans, family Familial periodic paralysis, polymyositis, inclusion body myositis, neuromyotonia, and stiff-man syndrome. In other embodiments, the myopathy becomes muscular atrophy, central axis disease, or malignant hyperthermia syndrome. In other embodiments, the muscular dystrophy is selected from the group consisting of Duchenne muscular dystrophy, facioscapulohumeral dystrophy, limb-type muscular dystrophy (limb) Girdle muscular dystrophy), myotonic muscular dystrophy, myotonic dystrophy type I, myotonic muscular, Bayesian muscular dystrophy (Becker's) Muscular dystrophy), congenital muscular dystrophy, distal muscular dystrophy, emery-Dreifuss muscular dystrophy, and pharyngeal muscular atrophy (Emery Dreifuss muscular dystrophy) 〇cul〇pharyngeal muscular dystrophy). In other embodiments, the glycogen storage disorder is selected from the group consisting of: p〇nipe, s disease, Andersen's disease, and comorbid (c〇H, s diseases). In other embodiments, the myoglobinuria is selected from the group consisting of McArdle's disease, Tau & Γ1ιί disease, and DiMauro disease. In other embodiments, the mitochondrial myopathy is selected from the group consisting of K e a r n s - S a y r e (Kearns-Sayre syndrome), MELAS and MERRF. In other embodiments, the individual is a non-human animal selected from the group consisting of: canine, equine, feline, porcine, murine, bovine, avian, and ovine. In an embodiment, the individual is a human. In another aspect, the invention provides a method of treating, preventing or ameliorating muscle fatigue in an individual comprising administering to the individual a therapeutically effective amount of a compound, the compound being described by a chemical formula selected from the group consisting of :SI, S2, S3, S4, S5, S6, S7, S9, S11, S12, S13, S14, S19, S20, S22, S23, S24, S25, S26, S27, S36, S37, S38, S40, S43 , S44 , S45 , S46 , S47 ' S48 , S49 , S50 , S51 , S52 , S53 , S54 ' S55 , S56 , S57 ' S58 , S59 , S60 , S61 , S62 - S63 ' S64 > S66 ' S67 ' S68 ' S69 'S70, S71, S72, S73, S74, S75, S76, S77, S78, S79, S80, S81, S82, S83, S84, S85, S86, S87, S88, S89, S90, S91, S92, S93, S94, S95' S96' S97, S98' S99' S100, 19 200815381 5101, S102, S103, S104, S107, S108, S 109, S110, Sill, S112, S113, S114, S115, S116, S117, S118, S119 , S120, S 1 21, S 1 2 2 and S 1 2 3, or any of the above-mentioned image isomers, non Like heterogeneous composition, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes, metabolites and prodrugs thereof. In certain embodiments, the compound is not S4, S7, S20, S24, S25, S26, S27 or S36. In one embodiment, muscle fatigue is muscle fatigue caused by exercise. In another embodiment, muscle fatigue is accompanied by any other disease or condition characterized by muscle fatigue. In one embodiment, muscle fatigue is caused by muscle lesions. In another embodiment, muscle fatigue is caused by muscle wasting. In another embodiment, muscle fatigue is caused by central axis air disease. In certain embodiments, the compound is selected from the group consisting of: S47, S48, S50, S51, S59, S64, S74, S7 5, S77, S85, S101, 5102, S103, S107, S109, S110, S111, S117 and S121, or any of the above image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes and prodrugs. In certain embodiments, the compound is selected from the group consisting of: S101, S102, S103, S107, S109, S110, si 11, si 17 and S 1 2 1, or any of the above-described mirror image isomers Non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes and prodrugs. In one embodiment, the compound is S 1 〇 7 . In certain embodiments, the individual suffers from exercise-induced muscle fatigue, age-related muscle fatigue, myopathy, neurological or neuropathic disease, infectious disease, chronic disorder, genetic disease, or any other 20 200815381 with A disease or abnormality associated with muscle fatigue. In certain embodiments, the individual is suffering from a heart disease or abnormality, an HIV infection, AIDS, muscular dystrophy, cancer, malnutrition, kidney disease, kidney failure, or a combination of any of the foregoing. In other embodiments, the individual suffers from an irregular heartbeat; an irregular heartbeat caused by exercise; congestive heart failure; chronic obstructive pulmonary disease; hypertension or any combination of the above. In certain embodiments, the individual's irregular heartbeat includes atrial and ventricular arrhythmia; atrial fibrillation and ventricular fibrillation; atrial frequency arrhythmia and ventricular pacing arrhythmia; atrial frequency and ventricular frequency; catecholamine polymorphic ventricle Frequency pulsation (CPVT) and the variation caused by the above motion. In some embodiments, exercise-induced muscle fatigue is caused by prolonged exercise. In other embodiments, muscle fatigue is caused by high intensity exercise. cause. In some embodiments, muscle fatigue is muscle fatigue caused by high intensity exercise in an individual. In another embodiment, muscle fatigue is caused by prolonged exercise. In other embodiments, muscle fatigue is age-related muscle fatigue in an individual. In one embodiment, muscle fatigue is accompanied by an abnormality in the disease. In certain embodiments, muscle fatigue is caused by muscle lesions. In other embodiments, muscle fatigue is due to certain aspects of muscular dystrophy in an individual, and the present invention provides a method of treating or preventing a muscle lesion in an individual, the method comprising administering to the individual a therapeutically effective amount of a compound'

The compound is a compound of the formula I, Ii, Ij, Ik, μ, 21 200815381 isomers, tautomers, tautomers, pharmaceutically acceptable treads, hydrates, solvates, complexes, metabolism Or prodrug or any combination of the above. In certain aspects, the invention provides a method of treating or preventing muscular atrophy in a subject, the method comprising administering to the individual a therapeutically effective amount of a compound of Formula I, Ia, Ib, Ic, id, & , 1 f 1 g, I_h, ii, y, b, h, b, Ι-η, I 〇 or ip, or its mirror image isomer, non-image isomer, tautomer, drug In combination with the above, a salt, a hydrate, a solvate, a complex, a metabolite, or any combination thereof. In certain aspects, the invention provides a method of treating or preventing a central disease in a subject, comprising administering to the individual a therapeutically effective amount of a compound of Formula 1, La, Ib, Ic, Id, Ie, If, Ig, u, u, 1 3 1虻, H, in, i_〇 or i_p, or its mirror image isomer, non-image isomer, tautomer, pharmaceutically acceptable Accepted salts, hydrates, solvates, complexes, metabolites or prodrugs or any of the foregoing, in certain aspects, the invention provides a method of treating or preventing malignant t in an individual, the method comprising administering to the individual A therapeutically effective amount of a compound which is a compound of Formula I, Ia, I_b, Ic, Id, Ie, If, Ig, Ih, τ · s 1 , Ij, I_k, 1-1, Im, In, Io or 卜p or not or its mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes, metabolites or Any combination of the above. In certain embodiments, the compound is

II b' Ie, if, i_g, u, 1-1, In, 1-〇 or ip represented by a substance or its mirror image isomer, non-image isomer, tautomer, 22 200815381 Pharma Acceptable salts, hydrates, solvates, complexes, metabolites or prodrugs or combinations of any of the foregoing. In other embodiments, the compound is selected from the group consisting of: Sl, S2, S3, S4, S5, S6, S7, S9, SI 1, S12, SI 3, S14, S19, S20, S22 , S23, S24, S25, S26, S27? S36, S37, S38, S40, S43, S44, S45, S46, S47, S48? S49, S50, S51, S52, S53? S54, S55, S56, S57, S58 , S59, S60, S61, S62, S63? S64, S66, S67, S68, S69, S70, S71, S72, S73, S74, S75, S76, S77, S78, S79, S80, S81, S82, S83, S84 , S85, S86, S87, S88, S89, S90, S91, S92, S93, S94, S95, S96, S97, S98, S99, S100, S101, S102, S103, S104, S107, S108, S109, S110, Sill , S112, S113, S114, S115, S116, S117, S118, S119, S120, S121, S122 and S123, and its mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, Hydrates, solvates, complexes, metabolites or prodrugs, and combinations of any of the foregoing. In certain embodiments, the compound is not SI, S2, S3, S4, S5, S6, S7, S9, S11, S12, S13, S14, S19, S20, S22, S23, S24, S25 as described herein. , S26, S27, S36, S37, S38, S40, S43, S44, S45, S46, S47, S48, S49 'S50, S51 'S52, S53, S54, S55, S56, S57, S58, S59, S60, S61 > S62 ' S63 ' S64 , S66 ' S67 , S68 , S69 ' S70 ' S71 ~ S72 ' S73 , S74 , S75 , S76 ' S77 , S78 , S79 , S80 ' S81 , S82 ' S83 , S84 , S85 , S86 , S87, S88 'S89, S90, S91, S92, S93, S94·, S95, S96, S97, S98, S99 or S100. In some embodiments of the application, the compound is not S4, S7, S20, S24, S25, S26, S27 or S36. In another embodiment, the compound is not jtv-519. In other embodiments, the method comprises administering to the individual a therapeutically effective amount of a compound represented by the formula In, 1-oxime or Ip, or a mirror image isomer thereof, a non-image isomer, a tautomer. A pharmaceutically acceptable salt, hydrate, solvate, complex, metabolite or prodrug or a combination of any of the foregoing. In other embodiments, the compound is selected from the group consisting of: S68, S101, S102, S103, S104, S107, S108' S109, S11, Sill, S112, S113, S114, S115, S116, SI 17. SI 18, SI 19, S120, S121, S122 and S123, and any of the foregoing salts, hydrates, solvates, polymorphs, complexes, metabolites, prodrugs and combinations thereof . In other embodiments 'the compound is selected from the group consisting of S 1 〇1, S 1 02, S103, S107, S109, S110, Sill, S117 and S121, and any of the above salts, hydrates , solvates, polymorphs, complexes, metabolites, prodrugs, in combination with any of the foregoing. In other embodiments, the compound is S107, any salt, hydrate, solvate, polymorph, complex, metabolite, peony, or any combination thereof. In other embodiments, the compound is S117, any salt, hydrate, solvate, polymorph, complex, metabolite 'prodrug, or any combination thereof. In certain embodiments of the methods of the invention, the compound is applied in the form of a particular salt, hydrate, solvate, isomer, mirror image isomer or substantially pure form. In certain embodiments, the compound is administered as a compound isolated in neat form on substantially 24 200815381, wherein the compound is S 1 17 . In certain embodiments of the method of treating myopathy or muscular dystrophy, the condition can be muscle fatigue, exercise-induced muscle fatigue, or any other condition associated with skeletal muscle defects. In another aspect, the invention provides a method of reducing calciin activity or plasma creatine kinase activity or content in an individual to treat or reduce skeletal muscle fatigue in an individual, the method comprising administering to the individual A therapeutically effective amount of a compound (represented by Structural Formula I). In another aspect, the invention provides a method of reducing calcitonin activity or creatinine kinase activity or content in an individual to treat or reduce skeletal muscle damage in an individual, the method comprising administering to the individual a therapeutically effective amount of a compound (Expressed by Structural Formula I). In another aspect, the invention provides a method of reducing calcitonin activity or plasma creatine kinase activity or amount in an individual to treat or reduce skeletal muscle function deficits in an individual, the method comprising administering to the individual a therapeutically effective amount of a compound (indicated by the structural formula). In another aspect, the invention provides for the reduction of intracellular calcium activating protease activity or blood creatine kinase activity or content to treat or delay muscle lesions such as, but not limited to, central axis disease or malignant hyperthermia syndrome. In the method, the method comprises administering to the individual a therapeutically effective amount of a compound (represented by Structural Formula I). In another aspect, the invention provides a method of reducing calcitonin activity or glycerol creatine kinase activity or amount in an individual to treat or delay the onset of muscular dystrophy in an individual, the method comprising administering to the individual a therapeutically effective dose Compound (represented by structural formula I). 25 200815381 In other embodiments, the compound is represented by the formula: Ι-η, 1-〇 or Ip ' or its mirror image isomer, non-image isomer, tautomer, pharmaceutically acceptable a salt, hydrate, solvate, complex, metabolite or prodrug or a combination of any of the foregoing. In other embodiments, the compound is selected from the group consisting of: SI, S2, S3, S5, S6, S9, S11, S12, S13, S14, S19, S22, S23, S37, S38, S40, S43, S44, S45 'S46, S47, S48, S49, S50, S51, S52, S53, S54, S55, S56, S57' S58, S59, S60, S61, S62, S63, S64, S66, S67, S68, S69, S70, S71, S72, S73, S74, S75, S76 'S77, S78, S79, S80, S81, S82, S83, S84, S85, S86, S87, S88, S89, S90, S91, S92, S93' S94, S95, S96, S97, S98, S99, S100, S101, S10 2, S103, S104, S107, S108, S109, S110, S111, S112, S113, S114, S115, S116, S117, S118, S119, S120 'S121, S122 and S 1 23, and the above-described mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes, metabolites or pre- Medicine and any combination of the above. In other embodiments, the compound is selected from the group consisting of S 1 0 1 , S 1 02, S 1 03, S107, S109, S1 10, SI 1 1 , S1 17 and S121. In one embodiment, the compound is S1 07 or a pharmaceutically acceptable salt thereof. In another embodiment, the compound is S117 or a pharmaceutically acceptable salt thereof. In certain embodiments, the compound is not SI, S2, S3, S4, S5, S6, S7, S9, SI 1, S12, S13, S14, S19, S20, S22, S2 3, S24, S25, S26 ' S27, S36, S37, S38, S40, S43, S44, S45 'S46, 26 200815381 S47, S48, S49 'S50, S51, S52, S53 'S54, S55, S56, S57, S58, S59, S60, S61, S62, S63, S64, S66, S67, S68, S69, S70, S71, S72, S73, S74, S75, S76, S77, S78, S79, S80, S81, S82, S83, S84, S85, S86, S87, S88, S89, S90, S91, S92, S93, S94, S95, S96: S97, S 9 8 , S 9 9 or s 1 0 0. In another embodiment, the compound is not s 4, S7, S20, S24, S25, S26, S27 or S36. In one embodiment, the compound is not JTV-519. In certain embodiments of the method of treating muscular dystrophy, the muscular dystrophy is selected from the group consisting of Duchenne muscular atrophy, facial scapula muscular atrophy, limb-type muscular dystrophy, and myotonic Muscular atrophy, Bayesian muscular dystrophy 'congenital muscular dystrophy, distal muscular dystrophy, Emery-Dreifuss muscular dystrophy and ocular pharyngeal muscular atrophy. In some aspects, the invention provides methods of treatment wherein the individual is a human. In certain embodiments, the individual is a non-human animal selected from the group consisting of: canine, equine, feline, pig, murine, bovine, poultry, sheep, drum, class, rodent Animals, sheep (〇vine) and bovidae. In other aspects, the invention provides methods of treatment wherein the compound is part of a pharmaceutical composition. In certain embodiments, the pharmaceutical composition comprises at least one pharmaceutically acceptable excipient. In other embodiments, the at least one pharmaceutically acceptable excipient includes aromatics, buffers, binders, colorants, 、^^^, 27 200815381 disintegrants, Diluents, emulsifiers, extenders, flavor-improving agents, gellants, glidants, preservatives, skin penetration enhancers (skin-penetration) Enhancers), solubilizers, stabilizers, suspending agents, sweeteners, tonicity agents, vehicles, viscosity-increasing agents ) or any combination of the above. In other embodiments, the pharmaceutical composition further comprises a second active formulation. In certain embodiments, the second active agent is an analgesic. In some embodiments, the composition is in the form of a capsule, a granule form, a powder form, a solution form, a suspension form or a tablet form or an animal feed form. In certain embodiments, the composition is administered by parenteral, enteral, intravenous, intraarterial, intraosseous, intraosseal, intracutaneous (intracutaneous), intracutaneous (intraosseal), intracutaneous (intracutaneous) ), subcutaneous, intradermal, subdermal, transdermal, intrathecal, intramuscular, intraperitoneal, intrasternal, parenchymal (intrasternal), parenchymal (intrasternal), parenchymal (intrasternal), parenchymal (intrasternal) Parenchymatous), oral, sublingual, buccal, rectal, vaginal, inhaled or intranasal. In certain embodiments, the compound is administered using a drug-releasing implant or via an osm〇tic pump. In certain embodiments, the subject is administered a dose of a compound that restores the binding of calstabin 1 to RyR1. In certain 28 200815381 embodiments, the individual is administered a compound sufficient to increase the dose of calstabin. In certain embodiments, the individual is administered a compound that reduces the amount of calcium-activated egg self-enzymatic activity. In some embodiments, the individual is administered a compound sufficient to reduce plasma creatine kinase activity or: a certain amount of fatigue caused by treatment stress or exercise: in the second, before the onset of exercise or stress ( 7, 6, 5, 4, 3, 2 in the process of simultaneous, at the end or after the end (1, 2, 3, 4, $ days),

The compound is administered. In certain embodiments, the dosage of 6:: is administered to the individual from about 1 mg/kg/day to about 2 mg: in certain embodiments, the dosage of the compound administered to the individual About gram; kg / kg / day to about i mg / kg / day. In certain aspects, the invention provides a method of treating or preventing muscle fatigue in an individual in need thereof, which comprises administering to the individual a therapeutically or prophylactically effective amount of a compound that reduces the rate of opening of the RyRl channel, in certain embodiments. The present invention provides a method of treating or preventing muscle fatigue in an individual towel having an effect, comprising administering to the individual a therapeutically or prophylactically effective amount of a compound (reducing Ca2+ ion flux through the RyR1 channel, in certain embodiments, The present invention provides a method of treating or preventing muscle fatigue in an individual in need thereof, comprising administering to the individual a therapeutically or prophylactically effective amount of a compound (reducible via

#5 ion leakage of the RyRl channel). In other embodiments, the invention provides a method of treating or preventing muscle fatigue in an individual in need thereof, comprising administering to the individual a therapeutically or prophylactically effective amount of a compound (which increases the affinity of calstabin j in combination with RyR 1 ). In other embodiments, the invention provides a method of treating or preventing muscle fatigue in an individual in need thereof, comprising administering to the individual a therapeutically or prophylactically effective amount of a compound (which reduces the separation of caistabin(R) from RyR1). In certain embodiments, the invention provides a method of treating or preventing muscle fatigue in a subject in need thereof, comprising administering to the individual a therapeutically or prophylactically effective amount of a compound (which may increase calstabin 1 recombination with RyRl). In certain embodiments, the invention provides a method of treating or preventing muscle fatigue in an individual in need thereof, comprising administering to the individual a compound that treats or prevents the effective agent 篁 (a combination of calstabin 1 and RyR l can be mimicked). In certain embodiments of the methods of the invention, the individual is suffering from a myopathy, a neurological disorder, an infectious disease, a chronic disease, a genetic disease, or any other disease or abnormality associated with symptoms of fatigue. Non-limiting examples of such diseases or abnormalities are heart disease or abnormalities, skeletal muscle function defects, HIV infection, AIDS, muscular dystrophy, cancer, malnutrition, muscle fatigue from exercise, age-related muscle fatigue, Kidney disease, kidney failure, irregular heartbeat; irregular heartbeat caused by exercise; jk-induced heart failure; chronic obstructive pulmonary disease; hypertension or any combination of the above. Individual irregular heartbeats include atrial and ventricular arrhythmia; atrial fibrillation and ventricular turnover; atrial frequency arrhythmia and ventricular pacing arrhythmia; atrial frequency and ventricular frequency; catecholamine polymorphic ventricular frequency (CpVT) and Variants caused by exercise. In some embodiments, the system is human. In other embodiments the system is a non-human animal, such as a canine, equine, feline, porcine, murine, bovine, poultry, sheep or any other animal in need of treatment. 30 200815381

The present invention relates to compositions and methods for treating muscle fatigue by administering a novel compound that stabilizes the Arnoxine receptor, which regulates calcium channel function in cells. The present invention provides a method of treating or preventing or reducing muscle fatigue by administering a guanidine, 4-benzothiazepine compound which is a RyR stabilizer. Individual muscle fatigue can be caused by many acute and chronic pathologies, diseases or symptoms, including but not limited to heart disease, HIV infection and AIDS, muscular dystrophy, cancer, malnutrition, exercise-induced muscle fatigue, age-related Sexual muscle fatigue, kidney disease and kidney failure. The present invention provides a method for treating or preventing about ion leakage and muscle fatigue of an individual's internal Anoplast body, wherein the individual string, a disease or abnormality characterized by muscle fatigue or the muscle fatigue of the individual is sustained and long Time and / or intense exercise or long-term stress. The present invention provides a method for the treatment of muscle fatigue by a calcium ion release channel stabilizer (for example, a RyCal compound). The present invention provides a method of reducing calpain activity 戋 a . The present invention provides a method of reducing plasma creatine kinase activity. V. This is the channel. The reduction in the amount used in the calcium ion release method inhibits the combination of the individual of the present invention. Individuals are fined to some compounds. The compounds used in other intracellular methods for intracellular methods can be modulated in the cells of the method of the invention. In other embodiments of the invention, the separation of the FKBP from the RyR is disclosed. The compounds can be used to enhance the individual methods of the RyR-FKBP complex of the present invention. In the compound-administerable example of the compound, the compound used in the pg method of the present invention for RyR is used in other embodiments of the compounds used in the intracellular FKBP and j. In other examples, the compound used in the method of the invention may be limited. Preventing or treating leakage of RyR receptors in an individual. In other embodiments, the compounds used in the methods of the invention modulate the binding of RyR to FKBP in an individual. In other embodiments, the compounds used in the methods of the present invention reduce the rate of RyR opening by increasing the affinity of FKBP for protein kinase A phosphorylated RyR. In some embodiments of the invention, the RyR is RyRl. In certain embodiments of the invention, the FKBP is calstabinl. In certain embodiments, the methods of treatment of the present invention are directed to methods of treating muscle fatigue caused by sustained or intense exercise. In other embodiments, the method of treatment of the present invention is directed to a method of treating muscle fatigue caused by prolonged stress. In certain embodiments, the invention relates to methods of treating or preventing muscle symptoms including, but not limited to, myopathy, such as muscular dystrophy. In certain embodiments, the invention relates to a method of treating a muscle lesion in an individual in need thereof, comprising administering to the individual a therapeutically or prophylactically effective amount of one of the compounds of the invention, such as Formula I, Ia, Ib, Ic a compound of Id, Ie, If, Ig, Ih, Ii, Ij, Ik, 1-1, Im, In, Io or Ip, or a mirror image isomer, a non-image isomer, a tautomer, A pharmaceutically acceptable salt, hydrate, solvate, complex or prodrug. The methods and compositions of the present invention can be used to treat any individual in need thereof. In certain embodiments, the system is a mammal, such as a mammal selected from the group consisting of: primates, rodents, sheep, bovidae, 32 200815381 pig, equine, feline, and canine. In the preferred embodiment, the system is human. The methods and compositions of the present invention can be used to treat or prevent symptoms, diseases, or abnormalities (e.g., myopathy) that affect muscle. For example, the methods and compositions of the present invention can be used to treat or prevent myopathy from a group consisting of: congenital muscle disease, muscular dystrophy, mitochondrial myopathy, endocrine myopathy, muscle Hepatic glycocalyx, myoglobinuria, dermatomyositis, muscle ossification, familial periodic itching, polymyositis, inclusion body myositis, neuromuscular rigidity and stiff human disease. A non-limiting example of a disease or abnormality affecting skeletal muscle function is central axis disease, malignant hyperthermia syndrome, etc. In a preferred embodiment, the methods and compositions of the present invention are useful for treating or preventing muscular dystrophy . For example, the methods and compositions of the present invention can be used to treat or prevent muscular atrophy selected from the group consisting of: Duchenne muscular atrophy, facial scapula muscular atrophy, limb-type muscular dystrophy, muscle Tonic-type muscular dystrophy, Bayesian muscular dystrophy, congenital muscular dystrophy, distal muscular atrophy, Emery-Dreifuss muscular dystrophy, and pharyngeal muscular atrophy. The compositions of the present invention can be administered by any suitable method known in the art. For example, the compositions of the invention may be administered by a route selected from the group consisting of parenteral, enteral, intravenous, intraarterial, intraspinal, intraosseous, intradermal, subcutaneous, intradermal, Subdermal, skin penetration, intrathoracic, intramuscular, intraperitoneal, intrasternal, parenchymal, oral, 33 200815381 4I ffl. Buccal, rectal, vaginal, inhaled or intranasal, $ use of music The implant is released by applying the implant. In the present invention, the preferred embodiment of the present invention relates to the treatment or prevention of muscle lesions in an individual in need thereof, the treatment of the individual or the treatment of the anti-injective agent. The compound S1〇7 U is like an isomer, a non-image isomer, a tautomeric wind, which is not a gas, hydrate, solvate, complex or prodrug. In a more preferred embodiment, the invention is related to::: Zhaozhong muscular atrophy (eg, Duchenne muscular dystrophy)::: need =: individual administration of a therapeutic or prophylactically effective amount of compound II or % , non-image isomers, tautomeric piles, salts, hydrates, solvates, complexes or prodrugs, which are attached to the [application] In the case of the patent application, the singular forms "a" and "the" are used in the singular and the singular Equals, and refers to "FKBP126 polymorphism, refers to one or more ΡΚΒΡ12.6 polypeptide (also known as ealstabin2) and the equivalents known to the skilled person, 胄, etc. All the public interest mentioned here U, patents and other references, the entire disclosure of which is incorporated herein by reference. Muscle fatigue can be caused by the following factors: intense or repetitive physical activity or symptoms; or any underlying physical symptoms of fatigue or ... fibrils and/or muscle function. Muscle fatigue is defined as sports tempering & + & failure - this can be assessed on a stress-type exercise test and defeated by a specific task (such as 咏^仃仃7杈Run/Treadmill running) Time quantification. The task of Φ sheng, ri, private failure is defined as the termination of exercise due to the inability to continue (this is defined as muscle fatigue).

\ Continuous or long-term motion is defined as performing motion beyond a specific and measurable time period. Intense exercise is a movement that causes muscle fatigue during a specific time period. Long-term stress is defined as a condition that causes long-term muscle fatigue, which is caused by sustained long-term exercise or stress, which is usually accompanied by long-term activation of the sympathetic nervous system (for example, "attack or escape (fight)慢性r Hight) "long-term activation of the reaction" caused by chronic diseases/abnormalities. In Example t, an individual's skeletal muscle function defect occurs during chronic obstructive pulmonary disease, hypertension, asthma, or hyperthyroidism. In certain aspects, the present invention relates to compositions and methods for treating and preventing myopathy. The term "my〇pathy" as used herein refers to a neuromuscular disorder caused by dysfunction of the muscle itself. The term "muscle lesion" as used herein includes all muscle lesions described herein and also includes all other muscle lesions known to those skilled in the art. 35 200815381 Muscle lesions may be hereditary (eg, many types of muscular dystrophy) or develop secondary musculoskeletal diseases and abnormalities including (but not limited to) congenital muscle lesions, muscle contractions (characterized by progressive muscle weakness), Granuloid myopathy, endocrine muscle disease, muscle glycogen storage, myoglobinuria, dermatomyositis, muscle ossification, familial periodic itching, polymyositis, inclusion body myositis, nerve Sexual muscle rigidity, stiff human disease, general muscle spasm (e〇mm〇n muscle cramps) and hand and foot spasm (tetany). Examples of muscular dystrophy include, but are not limited to, Duchenne muscular dystrophy, facial scapular limb muscle atrophy, limb-type muscular dystrophy, myotonic progressive muscular dystrophy, Bayesian muscular dystrophy, congenital Muscular atrophy, recurrent muscle atrophy, Emery-Dreifuss muscular atrophy and ophthalmic pharyngeal muscular atrophy. Examples of mitochondrial myopathy include, but are not limited to, Kearns-Sayre's disease, MELAS, and MERRF. MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke) is an abbreviation for "mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke", which is a progressive neurodegenerative disorder. MELAS affects a variety of organ systems, including the central nervous system (CNS), skeletal muscle, eyes, heart muscle, and the gastrointestinal and renal systems that are less likely to have a chance. MERFF (myoclonus epilepsy with ragged-red fibers) is an abbreviation for "myoclonus epilepsy with reddish fibers", which causes epilepsy (epHePsy), coordination loss, dementia, and muscle weakness. Examples of muscle glycogen storage include, but are not limited to, Pompe disease, Ander's disease, and Kohlosis. Examples of myoglobinuria include, but are not limited to, McCardel's disease, Tarre's disease, and Dharma's disease. 36 200815381 The definition of the same sink. Providing groups here

The vocabulary used herein is as follows: a preliminary definition of the terms used in this specification or a vocabulary, unless

The general formula I, I-a, I-b, l-c, id Μ, I-j, H, 1-1, Ι-η, n are referred to herein as "the compounds of the present invention". The compounds of the present invention are named using a digital nomenclature system, and compounds Nos. 1 to 123 are provided herein. Use the prefix "s" or the prefix "arm" to classify these numbered compounds. Therefore, the first numbered compound is called "S1" or "ARMO〇i", the second numbered compound is called "S2" or "ARM0 02", and the third numbered compound is called "S3" or " ARM003" and so on. The "S" and "ARM" naming systems can be applied interchangeably throughout the specification, illustration and patent application. The term "alkyl" as used herein means a straight or branched chain saturated hydrocarbon having 1 to 6 carbon atoms. Representative alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, second butyl (sec-butyl), tert-butyl, pentyl, isethyl Base, neopentyl, hexyl, isohexyl and neohexyl. The term "CrC4 alkyl" means a straight or branched alkane (hydrocarbon) group having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, and third. Butyl and isobutyl. 37 200815381 The term "r-alkenyl" as used herein means a straight or branched hydrocarbon having from 2 to 6 carbon atoms having at least ~ carbon-carbon double bonds. In one embodiment, an alkenyl group has one or two double bonds. The alkenyl group may exist in a ruthenium or osmium structure and the compound of the present invention comprises the above two structures. The term "alkynyl" as used herein means a straight or branched hydrocarbon having 2 to 6 carbon atoms and having at least one carbon-blocking triple bond. The term "aryl" as used herein means an aromatic group containing from 1 to 3 aromatic rings (not bite). The term "cyclic group" as used herein includes a cycloalkylheterocyclic group. The term "cycloalkyl" as used herein means a carbon ring of 3 to 7 members which is saturated or partially unsaturated. Any suitable position of the cycloalkyl group can be covalently attached to the defined chemical structure. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. The term "halogen" as used herein means fluorine, gas, bromine and iodine. The term "heterocyclic group" or "heterocyclic" or "heterocyclyl" or "heterocyclo" as used herein means at least one ring of carbon atoms. a fully saturated, partially unsaturated or fully unsaturated group having at least one hetero atom (eg, a single ring of 4 to 7 members, a bicyclic ring of 7 to 11 members, or a tricyclic system of 16 members), Each ring including an aryl group (i.e., "heteroaryl group") and which is bonded to a heterocyclic group containing a hetero atom of a heterocyclic ring 38 10 200815381 may have 1, 2, 3 or 4 selected from the group consisting of A heteroatom of a nitrogen atom, an oxygen atom and/or a sulfur atom, wherein the nitrogen and sulfur heteroatoms are selectively oxidized and the nitrogen heteroatoms are selectively quaternized. The heterocyclic group can be attached to the remainder of the molecule at any heteroatom or carbon atom in the ring or ring system. Exemplary heterocyclic groups include, but are not limited to, aZepanyl, azetidinyl, aziridinyi, dioxolanyl. , σ 南 south (furanyl), furaZanyl, homo piperazinyl, imidazolidinyl, imidazolinyl, isothiazolyl, Isoxazolyl, morpholinyl, oxadiazolyl, oxaz〇ndinyl, oxazolyl, orthoquinone. Oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, piperazinyi, piperidinyl, pyranyl , pyrazinyi, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyrido oxazolyl ), ° ratio σ 嗤 嗤 ( (pyridoimidazolyl), mouth ratio β 嗟 坐 坐 坐 (pyridothiazolyl), "pyridinyl", pyrimidinyl, pyrrolidinyl, "pyrrolidinyl" Pyrolinyl,. Quinuclidinyl, tetrahydrofuranyl, thiadiazinyl, thiadiazolyl, thienyl, 嗟 嗟嗤 39 39 200815381 (thienothiazolyl ), "thienooxazolyl", thienoimidazolyl, thiomorpholinyl, thiophenyl, triazinyi, and triazolyl (triaz〇iyl). Exemplary bicyclic heterocyclic groups include indolyl, isoindolyl, benzothiazolyl, benzoxazolyl, benzoxadiazolyl, and the like. , benzothienyl, (benzothienyl). qUinuClidinyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl σIndolizinyl, benzofuryl, benzofurazanyl 'chromonyl, coumarinyl, benzo-α-pyran Benzopyranyl, cinnolinyl, quinoxaliny 1, 17 indazolyl, pyrolopipyridyl, fluoropyridyl (such as fluorine [2] , 3-c]° thiol, fluoro[3,2-b]acridinyl or fluoro[2,3-b]acridinyl), dihydroisoindolyl, dihydropyridinium A group (for example, 3,4-dihydro-4'oxy-indolyl), three wells, triazinylazepinyl, tetrahydroquinolinyl, and the like. Exemplary tricyclic radicals include carbazolyl, benzidolyl, phenanthrolinyl, acridinyl, phenanthridinyl, or sulphate. (xanthenyl) and so on. The term "phenyl" as used herein means a substituted or unsubstituted stupid group. 40 200815381 The above terms "alkyl", "alkenyl", "alkynyl", "aryl", "phenyl", "cyclo", "cycloalkyl", "heterocyclyl", "heterocyclic" The heterocyclo) and "heterocycle" may further have one or more substituents as needed. Exemplary substituents include, but are not limited to, one or more of the following groups: hydrogen, halogen, CF 3 , hydrazine CF 3 , cyano, nitro, N 3 , keto, cycloalkyl, aryl, alkynyl , heterocyclic, aryl, alkaryl, heteroaryl or 0Ra, SRa, S( = 0)Re, S( = 〇)2Re, P( = 〇)2Re, S( = 0)20Ra, P( = 0) 2〇Ra , NRbRc , NRbS( = 0)2Re , NRbP( = 0)2Re, S( = 0)2NRbRe, P( = 0)2NRbRc, C( = 0)0Ra, C( = 0)Ra, C( = 〇)NRbRc, 0C( = 0)Ra, 0C( = 0)NRbRc, NRbC( = 0)0Ra, NRdC( = 0)NRbRc, NRdS( = 0)2NRbRc, NRdP( = 0)2NRbRc, NRbC ( = 0)Ra or NRbP( = 0)2Re, wherein Ra is hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, alkaryl, heteroaryl, heterocyclic or aryl; Rb, Re and Rd Is a hydrogen, an alkyl group, a cycloalkyl group, an alkylaryl group, a heteroaryl group, a heterocyclic ring, an aryl group, or, alternatively, Rb and Rc described above together with the nitrogen atom to which they are attached form a heterocyclic ring; Re is an alkyl group, a cycloalkyl group, an alkenyl group, a cycloalkenyl group, an alkynyl group, an alkylaryl group, a heteroaryl group, a heterocyclic group or an aryl group. Among the above exemplary substituents, groups such as an alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, a cycloalkenyl group, an alkylaryl group, a heteroaryl group, a heterocyclic ring and an aryl group may be optionally substituted by themselves. . Exemplary substituents may further comprise at least one calibration group, such as fluorescent, biologically luminescent, chemically luminescent, colored, and radiolabeled groups, as appropriate. The fluorescing group is selected from the following: dipyrrole methane boron difluoride compound (bodipy), dansone (dan"1), fluorescein, rhodamine 41 200815381 (rhodamine), Texas red (Texas red), cyanine dyes, pyrene, coumarins, Cascade BlueTM, Pacific Blue, Marina Blue ), Oregon Green, 4', 6-dimercaptobenzoquinone (DAPI), 11 bow I ° indopyra dyes, lucifer yellow, carbonized bite (propidium iodide), η than porphyrins, arginine, and variants and derivatives thereof. For example, ARM 1 18 of the present invention comprises a calibration group "Βς>DIPY", which is The main component is 4,4-difluoro-4·boron-3a,4a-diaza-s-hydrogen sulfoxide group (454-difluoro-4-bora-3a, 4a-diaza-<s,-indacene moiety) a family of fluorophores. For more information on the cursor-based group and fluorescence technology, see, for example, Richard P. Haughland's Handbook of Fluorescent Probes and Research C/zewica/s, 6th Edition (Molecular Probes, 1996) The entire text of which is incorporated herein by reference. Those skilled in the art can readily select an appropriate calibration base and incorporate the above-described calibration groups onto any of the compounds of the present invention without undue experimentation.

The term "quaternary nitrogen" as used herein means a tetravalent nitrogen atom, which includes, for example, a tetraalkylammonium group (for example, tetrakisammonium salt, N-methylpyridinium, N-methylpyridinium). Normal valence nitrogen; protonated ammonium species (eg, trimethyl-hydroammonium, N-hydropyridinium, n-hydropyridinium); N- Amine oxide (amine N ο X ides ) (for example, N-methyl-morphine-N-oxide 42 200815381 N-methyl-morpholine-N-oxide), N-oxidation port pyridine-N- The normal valence nitrogen in the oxide"; and the normal valence nitrogen in the N-aminoammonium salt (for example, N-aminopyridinium salt). Unless otherwise indicated, the nitrogen in the benzothiazepine ring of the compound of the present invention in the entire specification may be optionally a fourth-order nitrogen. Non-limiting examples thereof include ARM-1 13 and ARM-1 19. The compounds of the invention may exist in their tautomeric form, such as aini (ie) or imino ether. All such tautomeric forms are considered herein as part of the present invention. The term "prodrug" as used herein denotes a compound which, after administration to an individual, undergoes chemical conversion by metabolic or chemical processes to produce a compound of the invention. "Prodrug" means conversion to a prodrug in vivo ( Prodrugs of parent drugs are usually helpful because, in some cases, they are easier to administer than protoplasts. For example, they are bioavailable when administered orally, whereas prodrugs are not. The pharmaceutical composition also has two good solubility in the prodrug. For example, the compound carries a protective group which can be separated by hydrolysis in a body fluid (for example, 'blood) to thereby release the active compound' or the protective group in the body fluid. Is oxidized or reduced to release the compound. All stereoisomers of the compounds of the invention (for example, those due to various substituents The inclusion of a carbon atom and the presence thereof, including both the mirror image and the non-image isomer form, are considered to fall within the scope of the invention. For example, 43 200815381 individual stereoisomers of the compounds of the invention, which may be substantially Without other isomers (for example, pure or substantially pure optical isomers with specific activities), or may be a mixture, such as a racemic isomer or mixed with all other or other selected stereoisomers. The invention's ehiral centers may have S (left-handed) or R (right-handed) structures (as defined by "IUPAC 1 974 Recommendations"). Separation of racemic isomers by physical methods, such as Fractional crystallization; separation or crystallization of a non-figomereous derivative; or separation of chiral column chromatography by any suitable method from racemic isomers Individual optical isomers are obtained, which include, without limitation, conventional methods, such as salt formation followed by crystallization with an optically active acid. After preparation, it is preferably subjected to separation and purification to obtain a composition ("approximately pure" compound) containing a compound equal to or higher than 99% by weight, which is then applied as described herein. Or formulating the composition. The "substantially pure" I compound of the invention is also considered to be part of the invention. All structural isomers of the compounds of the invention may exist in a mixed, pure or large form. The U of the compound comprises cis (7): both trans-W-hydrocarbon isomers and the cis and trans of the ring or heterocycle. The term "metabolite" as used herein means in vivo (eg, chemistry in an individual) A by-product produced by the compound. 44 200815381 In the entire specification, groups and substituents can be selected to provide stable groups and compounds. The term "RyCal compound" as used herein means having the formula I, Ia (la), ib (lb), Ic (Ic), id (Id), Ie (Ie), If (If) provided by the present invention, Ig (Ig), ih (Ih), Ii (n), ij (ij), ik (Ik), Il (II), Im (Im), In (in), i-0 (I〇), Ip ( a compound of 1) or n, and is referred to herein as "a compound of the invention". The above compounds include, but are not limited to, any one or more compounds of the formula including, but not limited to, the following Compounds·· as defined herein, s 1, S 2, S 3, S 4, S5, S6, S7, S9, S11, S12, S13, S14, S19, S20, S22, S23, S24, S25, S2 6, S27, S36, S37, S38, S40, S43, S44, S45, S46, S47 > S48, S49, S50, S51, S52, S53, S54, S55, S56, S5 7, S58, S59, S60, S61, S62, S63, S64, S66 'S67 ' S68 ' S69 ' S70, S71' S72, S73 1 S74, S75, S76, S77, S78, S79, S80, S81, S82, S83, S84, S85, S86, S87 ' S88 ' S89 ' S90, S91, S92, S93, S94, S95, S96, S97, S98, S99, S100, S101, S102, S103, S104, S105, S107, S108, S109, S110, Sill, S112, S113, S114, S115, S116, S117, S118, S119, S120, S121, S122 and S1 2 3 . In certain embodiments, the compound is isolated and substantially pure. Individuals treated by the methods of the invention may include mammals. The above-mentioned mammals may include humans, primates, canines, equines, felines, pigs, murines, 45 200815381 bovine, poultry, ungulate or sheep. Here, the "animal", "individual" and "patient" include the animal kingdom and so on: it includes (but is not limited to) mammals, animals (for example, Jue, dog 'horse and humans.) pKA) "protein kinase A filled with acid" means the reaction of replacing the hydroxide with phosphate by the whitening of the enzyme.

\

"Back-phosphorylation" of the RyRl or RyR2 receptor means squaring by protein squamous A. A "pharmaceutical composition" means one or more of the pharmaceutically acceptable salts, hydrates, polymorphs or pre-treatments ', ^, 0 described herein. The purpose of the cardiopharmaceutical composition of a chemical component, such as a physiologically acceptable carrier and excipient, is to accelerate the administration of the compound to the organism. The compounds of the present invention may also be formulated as pharmaceutically acceptable salts such as the above acid addition salts and complexes. The above salts can be formulated to accelerate drug application by altering the physical properties of the agent without impeding its physiological effects. Examples of useful changes in the state of the object include, but are not limited to, lowering the melting point to accelerate the application of transmucosal and increase solubility to accelerate the administration of higher concentrations of the agent. The term "pharmaceutically acceptable salts" means salts which are suitable or compatible for the treatment of a patient or individual, such as a human patient or animal. 46 200815381 The term "pharmaceutically acceptable acid addition salt" as used herein means any of Formulas I, Ia, Ib, Ic, Id, Ie, If, ig, u, Ii, Ij, Ik, 1-1, Non-toxic organic or inorganic salts of the basic compounds represented by Im, In, Io or Ip or n or their intermediates. Exemplary inorganic acids which form suitable acid addition salts include, but are not limited to, hydrogen chloride, hydrobromic acid, sulfuric acid, and phosphoric acid, as well as sodium monohydrogen orthophosphate and potassium hydrogen sulfate (sodium monohydrogen orthophosphate). Potassium hydrogen sulfate). Exemplary organic acids which form suitable acid addition salts include, for example, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, Malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, benzoic acid, phenylacetic acid, cinnamic acid and salicylic acid, such as mono-, di- and dicarboxylic acids, and such as p-toluene acid ( P_t〇luene sUlfonic acid) and sulfonic acid such as methanesulfonic acid. Single or double acid salts can be formed and such salts exist in either of a hydrated state, a dissolved state or a nearly anhydrous state. In general, the acid addition salts of the compounds of the present invention are relatively soluble in water and many hydrophilic organic solvents, and acid addition salts generally exhibit a higher melting point relative to their free base type. The compounds of the invention may also be formulated as pharmaceutically acceptable salts, such as the acid addition salts and complexes described above. The above salts can be formulated to accelerate drug application by altering the physical properties of the agent without impeding its physiological effects. Examples of useful changes in the physical state include, but are not limited to, lowering the melting point to accelerate the application across the mucosa and increasing solubility to accelerate the administration of higher concentrations of the agent. 47 200815381 The term "pharmaceutically acceptable salts" means salts suitable for the treatment of humans, such as human patients. These salts can be ^^ I-a, I-b, I-c, Ld, I-e, I-f, I:g, I h, Μ, η, ! A non-toxic organic or inorganic salt of the compound represented by 彳, μ, I-m, Ι·η, I-0,][_p or any of the specific compounds described herein or any of the above intermediates. Exemplary salt-forming ions include, but are not limited to, ammonium ions (nh4 + ), calcium ions (Ca2+), iron ions (1^2+ and Fe3+), magnesium ions (Mg2+), and potassium ions (κ). +), acridine ion (C5H5NH+), quaternary ammonium salt (NR〆), sodium ion (: (Na), acetate, carbonate, chloride, bromide, citrate, cyanide, hydroxide, Nitrate, nitrite, oxide, phosphate, sulfuric acid hand, maleate, fumarate, lactate, tartrate, gluconate, besylate and propyl Illustrative inorganic acids which form suitable salts include, but are not limited to, hydrogen chloride, hydrobromic acid, sulfuric acid and phosphoric acid, and metal salts such as sodium monohydrogen phosphate and potassium hydrogen sulfate. Exemplary organic acids of addition salts include, for example, glycolic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, malic acid, tartaric acid, citric acid, ascorbic acid, cis Single or double with butyl phthalic acid, benzoic acid, phenylacetic acid, cinnamic acid and salicylic acid Acid protecting 'and sulfonic acids such as p-toluenesulfonic acid and methanesulfonic acid. Forming mono- or di-acid salts' and such salts are present in either hydrated, dissolved or almost anhydrous form. Said that compounds having the formula I, Ia, j_b, Bu "Ie, If, Ig, Bu h, Li, rj, u, Bu m, I n, Buddhism and Ip" are more soluble in acid addition salts. In water and many hydrophilic organic solvents, and with respect to their free base type, acid addition salts generally show a higher melting point in 48 200815381. Those skilled in the art can perform selection of suitable salts. For example, according to P · "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Heinrich Stahl and Camille G. or "Pharmaceutcial Salts" by Berge in j. pharm Sci, Vol 66 (1), p 1-19, 1977 To select salts. For example, other non-pharmaceutically acceptable salts (eg, oxalates) can be used for isolation or subsequent conversion to a pharmaceutically acceptable acid addition salt in a laboratory compound of the invention. Class I, Ia, Ib, Ic of the invention , Id, Ie, If, Ig, Ih, Ii, Ij, Ik, 1-1, Im, In, Io and I-ρ and compounds of formula ii may form hydrates or solvates, and this includes patent application Within the scope of the present invention, when the compounds of formula I of the present invention exist as regioisomers, configurational isomers, conformers, or diasteroisomeric Forms as well as the various mixtures described above are included within the scope of Formula I. If desired, individual isomers can be isolated by known separation and purification methods. For example, when the formula I of the present invention is a racemic isomer, the racemic isomer can be separated into the (S)-compound and the (R)-compound by optical resolution. Individual optical isomers and mixtures thereof are included in Formula I, Ia, Ib, Ic, Id, Ie, If, Ig, Ih, Ii, Ij, Ik, 1-1, Im, In, Io, and Ip, and In the scope of II. The term "solvate" as used herein means Formulas I, Ia, Ib, Ic, Id, Ie, If, Ig, Ih, Ii, Ij, Ik, 1-1, Im, In, Io 49 200815381 and Ip And a compound of formula II or a pharmaceutically acceptable salt of a compound of formula I wherein the molecular system of a suitable solvent is incorporated in a crystalline lattice. Suitable solvents are physiologically acceptable at the dosages employed. Examples of suitable solvents are ethanol, water, and the like. When water is a solvent, the molecule is considered a "hydrate." The term "polytype" means a specific crystalline state of a substance having specific physical properties (such as X-ray diffraction, infrared spectrum, melting point, etc.). The terms "effective dose", "sufficient dose" or "therapeutically effective dose" as used herein are sufficient to cause a beneficial or desired result (including clinical outcome), so the "effective dose" depends on the environment in which it is administered. . The response is for prevention and/or treatment. The term "effective dose" also includes a dose of a compound of formula I which is "effectively treated" and which avoids or substantially reduces the undesirable side effects. As used herein and as understood by the art, "treatment" is a method of obtaining beneficial or desired results, including clinical outcomes. Whether observable or unobservable, beneficial or desirable results may include, but are not limited to, amelioration or amelioration of one or more symptoms or diseases; reduction in the extent of the disease; stabilization (ie, no deterioration) of the condition of the disease; Prevent the spread of disease; delay or slow progression of disease; improvement and mitigation of disease status; and remission (whether in part or in whole). "Treatment" also means prolonged survival, as compared to the expected survival time if no treatment is received. The term "inhibiting isolation" as used herein includes hindering, reducing, inhibiting, limiting or avoiding the physical separation of FKBP subunits from RyR molecules in individual cells, as well as blocking, reducing, suppressing 丨ac inhibition, restriction or avoidance. RyR molecules within individual cells are physically separated or separated from the FKBP subunit. The term "enhanced binding" as used herein includes the ability to enhance, enhance or ameliorate the phosphorylation of RyR in human cells, such as κ, Ό σ FKBP (for example, a higher background than the negative control group). , about five times the binding capacity), and the ability to enhance, enhance or improve the physical binding of FKBp to phosphorylated RyR in individual cells (eg, the twist binding is about twice as high as the background binding of the negative control group). Five times the combined ability). The term "cardiomyocyte" as used herein includes myocardial fibers, such as those found in cardiac myocardium. The present invention provides a compound capable of treating and preventing abnormalities and diseases associated with the RyR receptor, which regulates the function of the mother ion channel in the cell. More specifically, the present invention provides a compound capable of treating or preventing leakage of a RyR channel. "Abnormality and disease associated with RyR receptors" means abnormalities and diseases that can be treated and/or prevented by modulating the RyR receptor (regulating the #5 ion channel function in cells). "Abnormalities and diseases associated with RyR receptors" include (but are not limited to) cardiac abnormalities and diseases, skeletal muscle abnormalities and diseases, cognitive abnormalities and diseases, malignant hyperthermia syndrome, central axis disease, diabetes and sudden infant death ( Sudden infant death syndrome). Cardiac abnormalities and disease burdens (but not limited to) irregular heartbeat abnormalities and diseases, irregular heartbeat abnormalities and diseases caused by exercise, sudden cardiac death, motor-induced sudden cardiac death, congestive heart failure , chronic obstructive pulmonary disease and high blood pressure. Irregular heartbeat abnormalities and diseases including irregular heart caused by exercise 51 200815381 Jump abnormalities and diseases including (but not limited to) atrial and ventricular arrhythmia; atrial fibrillation and ventricular fibrillation; atrial frequency arrhythmia and ventricular arrhythmia Atrial frequency and ventricular frequency; catecholamine polymorphic ventricular frequency (CPVT) and its motor-induced variants. Skeletal muscle abnormalities and diseases include, but are not limited to, skeletal muscle fatigue, exercise-induced skeletal muscle fatigue, muscle wasting, bladder dysfunction, and incontinence. Cognitive disorders and diseases include, but are not limited to, Alzheimer's Disease, various types of memory loss, and age-dependent memory loss. As referred to herein, the compounds of the present invention are capable of treating and preventing disorders and diseases associated with the RyR receptor (regulating intracellular calcium channel function) by accomplishing the following: repairing channel leakage and improvement Binding of FKBP protein (eg, calstabinl) to RyR phosphorylated by protein kinase A. Thus, in one embodiment, the compound is useful for the treatment and prevention of muscle fatigue associated with the RyR receptor, which regulates calcium channel function in cells. In one embodiment, the compounds of the invention are effective for treating muscle fatigue caused by myopathy, discomfort, disease, abnormality or symptoms associated with RyR receptors (regulating intracellular calcium channel function). Examples of such abnormalities and symptoms include, but are not limited to, heart disease or abnormalities, skeletal muscle function defects, HIV infection, AIDS, muscular dystrophy, cancer, malnutrition, exercise-induced muscle fatigue, age-related muscle fatigue, Kidney disease and kidney failure. 52 200815381 Examples of cardiac abnormalities and diseases include, but are not limited to, irregular heartbeat abnormalities and illnesses, exercise-induced irregular heartbeat disorders and diseases, congestive heart failure, chronic obstructive pulmonary disease, and hypertension. Examples of irregular heartbeat abnormalities and diseases include irregular heartbeat abnormalities and diseases caused by exercise, including (not limited to) atrial and ventricular arrhythmia; atrial fibrillation and ventricular fibrillation; , atrial pulse arrhythmia and ventricular frequency rhythm Incomplete; atrial frequency and ventricular stimuli; catechin polymorphic ventricular pulsation (CPVT) and its motor-induced changes In one embodiment, the compounds of the invention modulate about ion channels within an individual's cells. In another embodiment, the compounds of the invention reduce the release of the feeder ions into the cells. In another embodiment, the compounds of the invention reduce or avoid a reduction in the amount of FKBP bound to RyR in an individual. In another embodiment, the compounds of the invention inhibit the isolation of FKBP from RyR in a subject. In another embodiment, the compounds of the invention increase the binding of cells to RyR in an individual. In another embodiment, the compounds of the invention stabilize the RyR-FKBP complex in an individual cell. In another embodiment, the compounds of the invention prevent or treat leakage of RyR receptors in an individual. In the present invention, the compounds of the present invention modulate the binding of the ruler to FKBP in an individual. In another embodiment, the compounds of the invention reduce the rate of opening of RyR by increasing the affinity of fkbp for the phosphorylation of RyR of linsin kinase A. In another embodiment, the compounds of the invention reduce or inhibit calcium stimuli Chymotrypsin activity to treat muscle fatigue. In another embodiment, the compounds of the invention reduce blood creatinine kinase levels for the treatment of muscle fatigue. 53 200815381 The method of the invention can be practiced in vivo (Ζπ Wvo) or ex vivo (h vz7ro). Thus in one embodiment, the methods of the invention are practiced in an in vitro system (e.g., on isolated cell components in a test tube). In another embodiment, the methods of the invention are practiced in vivo, such as in cultured cells or tissues or within an individual. In another embodiment, the present invention provides the application of Formulas I, Ia, Ib, Ic, Id, Ie, i_f, Ι-g, ih, Ii, Ij, Ik, 1-1, Im, In, Io and 1- p and a compound represented by formula II for the preparation of a medicament for treating or preventing a muscle abnormality or disease such as, but not limited to, muscle fatigue in an individual. In another embodiment, muscle fatigue can be caused by increased pressure, such as an individual (e.g., a soldier or an athlete) who receives a sustained and prolonged exercise regimen. Thus in one embodiment, the compounds of the invention are used to treat muscle fatigue in an individual suffering from stress (e.g., due to, for example, a strong exercise regimen). Abnormal bone myopathy and diseases include, but are not limited to, stress-induced skeletal muscle fatigue, exercise-induced skeletal muscle fatigue, muscular dystrophy, bladder abnormalities, and fecal incontinence.

Pending applications USSN 1 1/2 12, 309 and 1 1/212, 413 and PCT application PCT/US2006/32405 teach the synthesis of a small number of lead compounds based on 1,4-benzothiazepines by Increase the binding of calstabinl to rescue RyRl channel function and Ca2+ leakage. Measuring the muscle fatigue 54 200815381

Defects in the Ca release channel (eg, the "leakage" of the channel can lead to fatigue of the bone path muscle. In some aspects, the present invention provides: 2 + a defect in a compound that can be used to treat, alleviate or Methods for preventing the following symptoms: muscles; muscle fatigue, including but not limited to muscle fatigue or muscle damage caused by exercise; muscle fatigue or injury associated with disease symptoms such as, but not limited to, muscle lesions, muscular dystrophy, and the like. Recent studies that lactic acid accumulation may not be harmful are triggering

Questions about the molecular mechanism of muscle fatigue. The hypothesis has proposed the role of 26 ion imperfect regulation. The present invention provides data showing changes in the function of the type 1 RyRi receptor during long-term exercise, and the main calcium ion release channel in the RyR1 skeletal muscle sarcoplasmic reticulum (SR) is coupled with excitatory contraction. (ECC) is required. Long-term movement during the RyRl channel in its Ser2844 (human

Ser2 8 43) is hyperphosphorylated by protein kinase a. The over-phosphorylation of protein kinase a is associated with the depleti〇n phosphodiesterase PDE4D3 in the RyRl complex. Furthermore, excessive acidification of protein kinase A causes the channel macromolecular complex to lack the RyRl stable subunit calstabinl (FKBP12), resulting in a "leakage" channel (increased opening rate of protein kinase A phosphorylation in the absence of normal channels) The degree of mobility and the lack of calstabinl and PDE4D3 are related to the intensity of exercise and rot and progressive fatigue. Both the mice lacking the calistal muscle calstabinl and the lack of PDE4D mice showed significantly impaired exercise capacity. The small molecule S107, which causes calstabinl to recombine in the RyR1 channel, improves the exercise capacity and strength of the isolated muscle during the 21-day exercise program. The intracellular ion can be measured by the repeated tonic contraction process. It is determined that S1 07 can treat muscle fibers and show a reduction in fatigue. Furthermore, si〇7 can treat long-term exercise in mice, which show plasma creatine kinase levels and calcium-dependent neutrality in muscle homogenates. Protease mom activates a decrease in protease activity. This verifies the existence of muscle fatigue mechanisms during prolonged or high-intensity exercise, Calcium ion signal defects and skeletal muscle damage can result from sarcoplasmic reticulum calcium ion leakage due to lack of calstabinl. In one aspect, the invention provides the use of a RyCal compound for muscle fatigue, muscle symptoms and Abnormal molecular mechanisms; and provide the above-mentioned treatment methods. #5Ion release channel stabilizing agent prevents muscle fatigue by avoiding calcium ion leakage of the Arino base receptor during continuous and intense exercise. Skeletal muscle is repeated and intense Activity can cause weakness (also known as fatigue) with strong application 2) pain and feeling of weak muscles (called perception), and 3) varying degrees of muscle decline (called atrophic reorganization (dystr〇phie rein〇deling) )). The dominant theory of muscle fatigue is that the accumulation of intracellular lactic acid causes intracellular acid 毋' to directly inhibit the production of muscle through fibrillin (Hill et al., 1929) below physiological temperature (2(TC), indeed found Changes in acidity of intracellular pH 可 promote skeletal muscle fatigue (Hill et al., 1929). However, more recent studies question the significance of acidosis to muscle fatigue, which shows repeated transient tonic contraction that causes muscle fatigue. In a more physiological state (37 ° C), there is no significant change in intracellular pH (Westerblad et al., 1997), in which acidosis does not significantly affect the production of force. The lactic acid is discharged at a basic rate via the lactic acid transporter (lactate 56 200815381 transporter). However, during the very intense athlete training, the lactic acid formed by the anaerobic cleavage of glycogen is still an important limiting factor. Importantly, it was found Intracellular acidosis can preserve muscle stress and myofibrillar relaxation during the long-term increase of intracellular Ca2+ during repeated or prolonged tonic contraction. 'Therefore avoiding muscle fatigue (Pedersen et al. 2004) 〇 Assume that intracellular pH 値 changes are not the main fatigue mechanism, which may cause fatigue in the changes in excitatory contraction coupling. Intracellular Ca2+ is triggered by the release of RyRl channels. Muscle contraction. In the current classical physiology experiments proposed in 1963, reversible changes in contractile activation may play an important role in muscle fatigue (Eberstein et al., 1963). In the process of repetitive tonic contraction leading to progressive development of fatigue, Decreased intracellular Ca2+ levels explain the reasons for the decrease in strength (Allen et al., 2001). However, caffeine and other compounds (which maximally activate the RyRl channel and cause sudden release of sarcoplasmic reticulum Ca2+) The tonic Ca2+ concentration can be transiently restored to normal (Allen et al., 2001). Therefore, changes in the RyRi-dependent sarcoplasmic Ca2+ release mechanism appear to be involved in fatigue formation. Measurement of sarcoplasmic reticulum Ca2 loading shows bone network A decrease in the total amount of Ca2+ released during muscle fatigue, which may be one of the causes of reduced sarcoplasmic reticulum Ca2+ release during fatigue (Cook e et al. 1 985). Another theory involves the concentration of intracellular inorganic phosphate ([Pi]〇 increases the precipitation of Ca2+ in the sarcoplasmic reticulum storage organ (Allen, 2001; Cooke, 1 985). The increase in [Pi]i (due to rapid breakage of ATP) occurs in the early stages of fatigue, but tonicity 57 200815381 The decrease in [Ca2+]i occurs in the late stage of fatigue (nith period). In addition, the increase of the stationary phase [Ca2+]i can be found after the retraction and straightening contraction, whereas the stimulation and tonicity [Cahh is reduced (Warren et ai 1993; (4) Xie e et ai 1 995). An increase in the static t-phase [Ca2+]i can cause long-term damage in the excitatory contraction coupling" by activation of a protease that can damage the Ca2+ release channel of the sarcoplasmic reticulum (La b et ai·, 1 995; Chln et Brut〇n, 1996) 〇r Long-term continuous exercise fatigue may result from sarcoplasmic reticulum Ca2+ leakage, which is caused by incomplete closure of the RyRJ channel, and sarcoplasmic reticulum “Η storage Partial disappearance contributes to the reduction in strength and the increase in quiescent period [(^2十]1, which interferes with the relaxation of muscles and when sustained employment will cause muscle decline. Recent data show that the pressure pathways that have been preserved by evolution, attack or escape response, Specific release of RyRl Ca2 + in skeletal muscle can be controlled (Gaburjak〇va et al., 2001; Marx et al., 2001), and abnormal, long-term activation of this stress pathway will trigger sarcoplasmic reticulum Ca2+ causing muscle fatigue. Leakage (Reiken et al., 2003) During the exercise, the RyRl/calcium ion release channel becomes over-acidified by protein kinase 且 and lacks the stable protein cal stab ini. The RyCal compound of the present invention enhances the calstabinl pair Protein kinase a hyperphosphorylation The binding affinity of RyR 1. These compounds are referred to as "calcium channel stabilizers" or "RyCal" and belong to the structure of 1,4-benzothiazepines and related structures. In a non-limiting example, RyCal compounds Treatment improves the motor performance of mice running on a treadmill. Furthermore, evidence suggests that calcium ion leakage through the proteinase A over 58 200815381 acid-filled RyRl channel can cause muscle damage due to activation of calcium-dependent proteases, RyCal can avoid #5 ion leakage and inhibit muscle damage during prolonged exercise. In certain embodiments, RyCal compounds can be used to treat, prevent, or ameliorate muscle fatigue in chronic conditions including, but not limited to, Heart failure, AIDS, cancer, septic, chronic obstructive pulmonary disease, hypertension, asthma, dysfunction of the squamous gland, chronic muscle fatigue. In other embodiments, RyCal compounds can be used to treat or ameliorate muscular dystrophy In other embodiments, treatment with a RyCal compound can prevent or reduce muscle fatigue, which can improve long-term sustained stress and/or In other embodiments, 'treatment with RyCal compounds can prevent or reduce muscle fatigue, which can improve the performance of individuals undergoing intense physical activity. The skeletal muscle is weakened due to intense use. For fatigue, in addition, stretch-dependent contracture can lead to additional muscle damage and decline. Although fatigue is seen as an important mechanism for limited peak performance and job failure during stress, The characteristics of the mechanism that causes fatigue or muscle fiber damage are not fully defined. In addition, the determination of fatigue molecular mechanisms can help prevent fatigue and muscle tissue damage (wehrens, 2005). An important physiological mechanism controlling skeletal muscle expression is the release of intracellular calcium (Ca2+), which is released from the Ca2+ specific storage site (sarcoplasmic reticulum) via the Argonine receptor (11>^111)〇32+. Released by the channel. In skeletal muscle, cell membrane depolarization activates voltage-gated L-type Ca2+ channels (LTCCs; Cavl.l), which in turn activates RyRJ on the sarcoplasmic reticulum by direct contact with the two ion channels. 59 200815381

The opening of the RyRl channel causes the release of a large amount of sarcoplasmic reticulum Ca2+, which activates myofibrils and muscle contraction. Furthermore, forms of disease associated with sustained activation of the sympathetic nervous system and increased plasma catecholamine levels can cause maladaptive activation of intracellular pressure pathways, resulting in instability of the RyR1 channel's closed state and intracellular Ca2+ leakage (Reiken et al) · 2003; Brillantes et al. 1994). It was found that sarcoplasmic reticulum Ca2+ leakage through the RyRl channel reduced calcium ion storage in the intracellular sarcoplasmic reticulum, increased compensatory energy consumption, and markedly accelerated muscle fatigue. Stress-induced muscle defects limit the pathology of the tip and contribute to muscle fatigue (permanently reducing performance). Furthermore, the instability of the RyRl shutdown state includes the lack of a stable channel subunit calstabinl (FKBP12) (Reiken et al. 2003 ϊ Brillantes et al. 19 94). Experiments have shown that increasing the binding affinity of calstabin to RyR restores channel function (Wehrens, 2003). The sarcoplasmic reticulum can activate skeletal muscle contraction via Ca2+ release from type 1 skeletal muscle Ano receptor (RyRl). Depolarization of the transverse tubular membrane activates the voltage inducer of the dihydropyridine receptor (Cavl.l), which in turn activates the RyRl channel via direct protein-protein interaction resulting in the release of sarcoplasmic reticulum Ca2+ storage . Ca2+ binding to troponin C causes the actin-myosin cross-bridge to produce and shorten the sarcomere. The Ca2+ release channel comprises a macromolecular complex consisting of a 560 kDa RyRl subunit homotetramer that forms the backbone of regulatory channel functional proteins, including: protein kinase A and phosphodiesterase. PDE4D3, both bound to the channel via the mosaic protein mAKAP; ppi (via spinophilin binding); and calstabinl (FKBP 12) 60 200815381 (Jayaraman, Brillantes et al. 1 992; Brillantes, Ondrias et al. 1994; Marx, Reiken et Al. 2000; Marx, Reiken et ah 2001) The stimulating contraction coupling defect that causes the sarcoplasmic reticulum Ca2+ release amplitude to decrease will impair contraction and force production. Eberstein and Sandow suggest that the release of damaged sarcoplasmic reticulum Ca2+ may be a contributing factor to muscle fatigue (Eberstein and Sandow 1963). It has been described that fatigue stimulation can cause a decrease in the amplitude of sarcoplasmic reticulum Ca2+ release (Allen, Lee et al. 1989; Westerblad and v Allen 1991; Allen and Westerblad 2001). In addition, the decline in Ca2+ storage during intense and repetitive contraction has been shown (Kabbara & Allen, 19 99), and it has been found that the time course of self-fatigue recovery is similar to the time course of long-term inhibition of sarcoplasmic reticulum Ca2+ release. (Westerblad, Bruton et al. 2000). Furthermore, it has been suggested that the precipitation of inorganic calcium ion phosphate during fatigue can cause a decrease in free Ca2+ in the sarcoplasmic reticulum (Allen and Westerblad 200 1). Evidence for the release of sarcoplasmic reticulum Ca2+ release from fatigue muscles. The role of RyR1 regulated sarcoplasmic reticulum Ca2+ release in fatigue. The combination of Calstabinl (FKBP12) on RyRl stabilizes the closed state of the channel and promotes coupling between adjacent channels (Brillantes, Ondrias et al. 1994; Marx, Ondrias et al. 1 998). Removal of calstabinl from drug RyRl (using rapaniyCin or FK506, both of which binds to calstabinl and separates it from the RyR1 macromolecular complex) can cause sub-conducting states and can be found in intact skeletal muscle. The depolarization caused the rapid contraction of 61 200815381 (Lamb and Stephenson 1996). RyRi mutations that cause loss of calstabinl binding can result in impaired excitatory contraction and maximum reduction in Ca2+ release from voltage-gated sarcoplasmic reticulum without affecting the storage of sarcoplasmic reticulum Ca2+ (Aviia, Lee et al. 2003) ). Hereditary removal of FKBP12 in skeletal muscle can cause severe tissue or developmental defects, but severe cardiac development defects are found, a detailed assessment of unicorn skeletal muscle function (Shou, Aghdasi et al. 1 998). Conversely, specific knock-out of skeletal muscle results in a decrease in Ca2+ release from the voltage-controlled sarcoplasmic reticulum and increases L-channel currents in isolated myotubes (Tang, Ingalls et al. 2004). In the extensor digitalis longus (EDL) rather than the soleus (soleus) or diaphragm, the reduction in maximum maximal straight force and the right-off shift in force-frequency relationship can be found (Tang, Ingalls et al. 2004). ). These data indicate that calstabinl regulates the acquisition of excitatory contraction coupling in skeletal muscle.

The binding of Calstabinl to RyRl is regulated by phosphorylation of protein kinase A in RyRl-S2 843 (position S2844 in the mouse RyRl sequence) (Reiken, Lacampagne et al. 2003). Phosphorylation on RyRl-S2843 increases the average opening rate of RyRl in the lipid bilayer (Reiken, Lacampagne et al. 2003). The RyRl-S2843A mutant channel is not phosphorylated by protein kinase A and therefore does not show the same protein kinase A-dependent increase in the rate of opening. The RyRl-S2843D mutation that mimics protein kinase A phosphorylation has a high onset rate and has an unstable on and off state (Reiken, Lacampagne et al 1. 2003). Since other research groups have found no effect on channel function or 62 200815381, the role of protein kinase A phosphorylation in RyRl is still being investigated (Stange, Xu et al. 2003). Other post-translational modifications of RyRl have been shown to modulate the binding of calstabinl to RyRl, including oxidative and glutathionylation of up to 50 free (reduced) thiols on each RyR monomer. . It has been confirmed in the myofibrils and muscle atrophy models after intense exercise that the tendon network Ca2 leakage is an abnormal #5 ion spark (Wang, Weisleder et al. 2005), which may be functioned by RyRl. The long-term activation of the sympathetic nervous system (SNS) in heart failure is related to: early skeletal muscle fatigue; and protein kinase A hyperphosphorylation on RyRl Ser2844 (meaning skeletal muscle in heart failure, The average of four protein kinase A positions per homotetramer channel is 3-4 via protein kinase A; the RyRl complex lacks calstabinl; and increased functional channel defects (Reiken, Lacampagne et al. 2003) RyR1 dysfunction in skeletal muscle can lead to localized subcellular Ca2+ release in variants (Ward, Reiken et al. 2003) Modifications in the 〇I RyR 1 complex can change during prolonged or high-intensity exercise and may Limiting muscle tip performance, increasing muscle fatigue and contributing to muscle damage. In some aspects, the present invention provides long-term, high-intensity, forced-moving mouse mode applications to assess RyR1 channels Role in skeletal muscle fatigue. As described herein, the RyRl channel macromolecular complex recombines during exercise such that it is progressively over-phosphorylated by protein kinase A and lacks PDE4D3 and caistabinl. Functionally, this recombination is accompanied 63 200815381 The "leakage" channel (increased opening rate) and the activation of an ion-activated protease (partial activating protease) and the leakage of creatine kinase (cPK) into plasma are consistent with muscle damage. Reduced and impaired exercise capacity in isolated muscles and worsened in mice lacking PDE4D or muscle specificity lacking calstabinl. Prevention of RyRl channel leakage with #5 ion channel stabilizer S107, which can be enhanced Caistabin1 binds to RyR 1, inhibits the activation of calpain and creatinine kinase, and improves motor performance. Thus, it causes leakage channels, activation of ion-activated proteases (calcium-activated proteases), and Creatine kinase (CPK) leaks into the plasma RyRl channel complex recombination, which is involved in long-term or high-intensity exercise Mechanism of meat fatigue. Confocal focal image studies of intracellular Ca2+ release (Ca2+ spark waves) in muscle cells after mild and intense treadmill exercise show that fatigue movement causes abnormal Ca spark wave activity (Wang et al· 2005). In addition, myofibrils with abnormal Ca2+ spark wave activity (caused by intense exercise) show signs of a decline in toxic Ca2+ effect (also known as "dystrophic remodeling") (Wang et Al· 2005). Sustained exercise for several weeks and months may cause abnormal RyRl function, intracellular Ca2+ leakage, decreased muscle performance, and muscle atrophic reorganization. Furthermore, 1,4-benzothiazepines improve muscle tip performance and prevent atrophic recombination by correcting intracellular Ca2+ leakage caused by stress. Animal models can determine muscle fatigue and atrophy caused by defects in RyRl function in vivo, isolated skeletal muscle cells, and a single RyRl channel. The use of these fatigued animal models and characteristic muscle, cell and channel techniques allows for trials based on modified RyR 1 leakage treatments that can improve skeletal muscle performance, muscle fatigue reduction and atrophy during prolonged exercise. Reduction of sexual reorganization. Muscular atrophy Type 1 atrophic myotonia (DM1), the most common muscular atrophy in adults (1 in 7,400 neonates), a myotonic dystrophy protein kinase (myotonic dystrophy protein kinase) , DMPK) The 3' not I/translated region of the CTG trinucleotide repeat amplification caused by multiple systemic disorders, which cause progressive muscle weakness, hereditary muscle hyperexcitability (muscle rigidity) Cardiac conduction defects, cataract and insulin resistance (Bachinski LL, Udd B, Meola G, et al. Confirmation of the type 2 myotonic dystrophy (CCTG) n expansion mutation in patients with proximal myotonic myopathy/proximal myotonic dystrophy of Different

European origins: a single shared haplotype indicates an I ancestral founder effect. Am J Hum Genet. Oct 2003;73(4):83 5-848. Hamshere MG,Harley H,Harper P, et al. Myotonic dystrophy: the correlation of (CTG) repeat length in leucocytes with age at onset is significant only for patients with small expansions. J Med Genet, Jan 1 999; 3 6(1): 5 9-6 1 ; Liquori CL, Ricker K, Moseley ML, et Al. Myotonic dystrophy type 2 caused by a 65 200815381 CCTG expansion in intron 1 of ZNF9. Science. Aug 3 2001; 293 (553 1): 864-867). Mutant DMPK signaling RNA (mRNA) containing a repetitive CUG expansion is retained in the nucleus and thus reduces its protein content (Mankodi A, Logigian E, Callahan L, et al. Myotonic dystrophy in transgenic mice expressing an expanded CUG repeat. Science. Sep 8 2000; 289 (5485): 1769-1773). Repeated expansion of RNA alters chromatin structure; silencing of side genes (encoding transcription factors); and disrupts regulation of gene expression during development and exercise (Ebralidze A, Wang Y, Petkova V, et al. RNA leaching of transcription Factors disrupts transcription in myotonic dystrophy. Science. Jan 16 2004;303(5656):383-387) In the type 1 atrophic myotonia, the cause of most serious symptoms (including muscle weakness and progressive muscle weakness) seems to be Increased by intracellular Ca2+ concentration and concomitant myofibrillar decay (Jacobs AE, Benders AA, Oosterhof A, et al. The calcium homeostasis and the membrane potential of cultured muscle cells from patients with myotonic dystrophy. Biochim Biophys Acta. Nov 14 1990; 1096(1 ): 14-1 9). Furthermore, recent studies have established a correlation between the Ca2+ cycle disturbed in Type 1 atrophic myotonia and the abnormal splicing of RyRl and sarcoplasmic reticulum Ca2+ ATPase (SERCA1) mRNA (Kimura T? Nakamori M? Lueck JD, et al. Altered mRNA Splicing of the Skeletal Muscle Ryanodine Receptor and Sarcoplasmic/Endoplasmic Reticulum Ca2 + -ATPase in 66 200815381

Myotonic Dystrophy Type 1 . Hum Mol Genet· Jun 2 2 2005). The muscle-specific gene model of type 1 atrophic myotonia can survive, and the performance of repeated expansion of CUG results in a phenotype similar to atrophic myotonia (Mankodi A, Logigian E, Callahan L, et al· Myotonic dystrophy in transgenic Mice expressing an expanded CUG repeat. Science· Sep 8 2000; 28 9(5 4 8 5): 1769-1773). In the absence of muscle fiber necrosis, there is a phenotype of muscle rigidity, while short and long forms are repeated! Mouse strains show relatively few or more muscle regeneration and repair groups κ * woven signs (Mankodi A, Logigian E, Callahan L, et al. Myotonic dystrophy in transgenic mice expressing an expanded CUG repeat. Science· Sep 8 2000; 289 (5485): 1769-1773). Since mice with short or long repeat performance do not show signs of muscle weakness, and because of the association between RyRl changes and type 1 atrophic muscle strength, ugly "LR mice provide research exercise and sympathetic nerves The system activates the pattern of effects in these mice. After describing the RyR 1 channel composition, phosphorylation status and functional characteristics, the 'Continuous Exercise Test and RyCal Compound Treatment can be used to stimulate the ugly 1111 mice. A type of atrophic myotonia with a transcript of repeated expansion CUG (the transcript) causes a mechanism of muscle rigidity and muscle decline 'This will provide 1) a study of animal models of severe fatigue, 2) type 1 atrophic myotonia Description of the mechanism, and 3) The basic theory of the development of treatment for muscle atrophy common in adults.. Pressure pathway 舆 ▲ ▲ meat 67 67 67 200815381 、 墼 途 么 的 的 , , , , , , , , , , , , , , , , , , , , Such as, but not limited to, fighting) can cause muscle loss and/or woven damage. Due to the sustained high content of catecholamine (causing intracellular Ca 2 + leakage) toxicity is the main determinant of muscle damage. This idea can be supported by 歹〗 1) Physiological (non-fighting) exercise or stress can be in sensitivity ~ 丨 仏 仏 肌 肌(rhabdomyolysis)) causes muscle weakness and tissue atrophy (Wappler et al., 2001); 2) intense and T is a significant C in the frequency of Ca2+ spark waves in muscle cells caused by gentle treadmill movement; Ca2+ leakage (Wang et al., 2005); 3) Excessive catecholamines found in malignant hyperthermia syndrome (MH) and central axis airway disease (CCD) cause uncontrolled release of intracellular Ca2+ (Μ 〇ηη^Γ to 2〇〇〇;

MaeLennan et al. 1995); 4) Most malignant hyperthermia/central axis disease is associated with RyRl missense mutations (Loke et aU 2〇〇3); 5) as high adrenaline states occur in heart failure ( Hyperadrenergic state) can cause RyRl hyperphosphorylation, Ca2+ leakage and skeletal muscle fatigue (Reiken et al. 2003); and 6) by activation of adrenergic receptors, intracellular cAMP production and protein kinase Α phosphorylation Excessive plasma catechol V amine can cause muscle damage (Goldspink et al., 2004; Tan et al. 2003). Pressure-dependent muscle damage and dysfunction occur at the junction between the intracellular catechol phenolamine effector (protein kinase A, PKA) and intracellular Ca2+ release. The ossicular alkaloid receptor (RyR1) Cah release channel constitutes an intracellular skeleton that combines protein kinase A-mediated stress signals with intracellular Ca2+ release regulation, thus determining excitatory contraction coupling and muscle function. Obtained. Importantly, the long-term increase in long-term high adrenaline status in vivo 68 200815381

Phosphorylation of RyRl protein kinase A will lack the stability of the calstabinl subunit ^ into the muscle network Ca2+ leakage and muscle fatigue. Furthermore, these findings can be extended to fatigue animal models in vivo. Therefore, sustained activation of the sympathetic nervous system during sustained muscle performance contributes to an increase in fatigue development and atrophic skeletal muscle injury. Since the skeletal muscle RyRl Ca2 + release channel is excessively sulphated by protein A and lacks the stable calstabinl subunit after 21 days of intense exercise, similar changes in the pot can cause intracellular Ca2+ leakage in heart failure animals. So long (more than 1 week) form of exercise can cause harmful intracellular Ca2+ leakage (via defective RyRl channels). In one aspect, the present invention establishes a molecular mechanism by which muscle fatigue occurs over a long period of time. In another aspect, the invention provides a method of treating or preventing deleterious intracellular Ca2+ leakage and muscle damage by administering a novel 1,4-benzothiazepine derivative. The skeletal muscle anodine receptor (RyR1) channel system forms a macromolecular signal complex from four RyRl subunits and related proteins (binding to the cytoplasmic region of the channel). Certain aspects of the invention examine the mechanism by which the function of RyR1 is regulated via an allosteric modulator. Examination of two specific forms of ectopic regulation\1) eAMp-dependent protein kinase A (PKA) regulation of the channel, which effectively activates channel gating; 2) long-term activation of the sympathetic nervous system (due to intense, sustained motion) When the protein kinase A phosphorylation is increased for a long time, the RyRl channel lacks the stable subunit calstabinl. RyRl's protein kinase A dysregulation during continuous exercise can cause intracellular Ca2+ leakage and may be a mechanism for increased muscle fatigue and atrophic recombination. In some aspects, the present invention measures the effects of fatigue on the following: 1) RyRl protein kinase A acid cleavage; 2) RyRl channel function, assayed by reconstituting RyRl channels in a double lipid membrane; 3) Calstabinl Binding to RyRl; 4) intracellular Ca2+ spark waves in isolated myofibrils; 5) isolated skeletal muscle function; 6) granular gland integrity; 7) in vivo motor performance; 8) skeletal muscle Histology, fiber composition type and oxidative capacity; 9) Creatine kinase (CK) plasma content; 10) Protein kinase A structuring of RyR1 in white blood cells and lack of calstabinl. Calstabinl deficiency and prolonged phosphorylation of protein kinase A cause prolonged activation of RyR1, intracellular Ca2+ leakage, decreased sarcoplasmic reticulum Ca2+ storage, accelerated fatigue, and muscle atrophic reorganization. In certain embodiments, treatment with any RyCal compound can normalize RyR dysfunction and intracellular Ca2+ leakage. In heart failure mode and intense exercise, RyRl is hyperphosphorylated by protein kinase A (PKA) and is "leakage". RyRl can recombine calstabinl after treatment with RyCal compounds, a single channel in heart failure. Normalize and improve muscle performance. RyCals may prevent intracellular Cah leakage during continuous exercise and normalize RyRl channel function in cells treated with RyCais in vivo. By preventing Ca2+ leakage from RyRl, RyCal compounds can improve skeletal muscle fatigue and atrophic recombination of Y during intense exercise (like fighting) by applying two muscle-fatigue animal models in mice and rats (swim with: Running on the car, some aspects of the invention show that treatment with RyCal compounds can be pre-70 200815381 anti-RyRl lacks calstabinl, which reduces muscle fatigue, improves performance and inhibits muscle atrophic reorganization. The molecular mechanism of muscle fatigue in Type 1 atrophic myotonia is confirmed in certain aspects of the present invention. In certain embodiments, the Ry Cal compound improves skeletal muscle fatigue time and atrophic recombination in atrophic myotonic mouse mode by preventing Ca2+ leakage of RyRl. Other aspects of the invention describe the characteristics of the molecular mechanism of fatigue in a mouse model of genes. In certain embodiments, calstabinl and PDE4D3 knockout mice can be applied to further explore the fatigue molecular mechanisms and dysfunctions that cause intracellular Ca2+ leakage during sustained motion. In other aspects, the compounds of the invention reduce the activity of calcium activating proteases. In other aspects, the compounds of the invention reduce plasma creatine kinase activity. Other aspects of the invention characterize the mechanism by which the intracellular calcium ion (Ca2+) release channel is unstable during long-term activation of the sympathetic nervous system (as occurs in continuous motion and/or combat). The skeletal muscle anodine receptor (RyRl) has two ectopic regulators, one being protein kinase A and the other being the channel-stable protein subunit (calstabinl) (Wehrens et al., 2004). Other aspects of the invention disclose therapeutic and prophylactic measurements in which an agent that recombines calstabinl prevents skeletal muscle fatigue by normalizing the skeletal muscle aromatine receptor (RyRl). It is known that the hyperphosphorylation of protein kinase A (PKA) of RyR1, which causes sarcoplasmic reticulum ca2+ leakage, is one of the causes of fatigue during long-term activation of the sympathetic nervous system (Reiken et al.? 2003), and this phenomenon can be reversed by JTV519 (Reiken et al.? 2003) Wehrens et al, 2〇〇5). 71 200815381 In a second aspect, the present invention provides: a description of RyRl Ca2 + leakage 70 characteristics of intense exercise, in vivo and ex vivo (ex Wvo) muscle performance and metabolism therein; use of RyCal compounds to overcome muscle fatigue and is intense And/or methods of avoiding muscle damage and/or fatigue during exercise, and methods of preventing muscle damage and/or fatigue associated with abnormal skeletal muscle function or any disease symptoms using y chemical & Certain aspects of the invention characterize the mechanism by which the intracellular calcium ion (Ca2+) release channel is unstable, which is necessary for muscle relaxation and avoids uncontrolled intracellular sarcoplasmic reticulum Ca2+ leakage damage. Long-term activation of the sympathetic nervous system during forced and sustained exercise caused RyRl's protein to over-cyanate, lack of calstabinl and channel closure state defects associated with sarcoplasmic reticulum Ca2+ leakage. Since the stress and physical performance in combat are significantly more severe than those in animal use, it can be inferred that the muscle fatigue and atrophic decline caused by Ca2+ leakage show a more serious phenotype among the fighters. One of the two ectopic regulators of the skeletal muscle alkaloid receptor (RyR1) is protein kinase A (which is an important stress pathway) and the other is the channel-stable protein subunit (calstabinl). Some aspects of the present invention suggest potential therapeutic and prophylactic measurements in which clastabinl recombined agents can prevent skeletal muscle fatigue by normalizing skeletal muscle receptors (RyRl). It is known that when the sympathetic nervous system is activated for a long time, the hyperphosphorylation of protein kinase A (PKA) of RyR1 causing sarcoplasmic reticulum Ca2+ leakage is one of the causes of fatigue (wehrens XH, Lehnart SE, Reiken S5 et Enhancing calstabin binding 72 200815381 to Ryanodine receptors improves cardiac and skeletal muscle function in heart failure. Proc Natl Acad Sci USA· Jul 5 2005;102(27):9607-9612 ; Ward CW? Reiken S,

Marks AR, et al. Defects in ryanodine receptor calcium release in skeletal muscle from post-myocardial infarct rats. Aug 2003; 17(1 1): 1517-1 519). Some aspects of the invention are described in terms of the full characterization of the effects of intense exercise on RyRl Ca2+ leakage; in vivo and in vitro (d W v ο) muscle performance and energy metabolism; and the use of RyCal compounds to improve muscle fatigue time and exercise Ability and prevention of intense, sustained exercise and/or subsequent sports injuries. The quality of life and prognosis of patients with heart failure are reduced by skeletal muscle dysfunction (eg, shortness of breath due to weak diaphragm muscles and exercise intolerance due to muscle fatigue in the quadrilateral muscles) (Harrington et al. 1997). Recent studies have confirmed that intracellular Ca2+ release (from sarcoplasmic reticulum) is a pathological mechanism that causes skeletal muscle dysfunction in heart failure (Reiken S, Lacampagne A, Zhou H, et al. PKA phosphorylation activates the calcium release). Channel (ryanodine receptor) in skeletal muscle: defective regulation in heart failure. J Cell Biol· Mar 17 20 03; 160(6): 919-92 8.; Ward, 2003; Perreault CL,

Gonzalez -Serratos H, Litwin SE, et al. Alterations in contractility and intracellular Ca2+ transients in isolated bundles of skeletal muscle fibers from rats with chronic heart failure· iies· Aug 1 993 ;73 (2) :405-4 1 2). Myocardial infarction 73 200815381 Heart failure in myocardial infarct animals causes significantly accelerated fatigue, which is inherent in fatigue skeletal muscle and measured in a manner that the tonic force is less than 50% of the maximum contractile force (Reiken, 2003). Skeletal muscle RyR1 is hyperphosphorylated by protein kinase A and lacks calstabin1. Previous studies have demonstrated that skeletal muscle RyR1 channel is over-acidified by protein kinase A in rat model of heart failure induced by rhythm and in rat model of myocardial infarction Calstabinl (Reiken et ai 2003; Ward et al., 2003). Furthermore, in the mouse model of heart failure after myocardial infarction, the RyR1 line in the soleus muscle is hyperphosphorylated by protein kinase A. In certain embodiments t, treatment with a RyCal compound allows calstabinl to recombine with RyRl (even in intense prolonged exercise). When exercise or stress (attack or escape response) requires improved muscle performance, no-adrenalin stimulation increases the excitatory contraction coupling obtained. The catechol phenolamine binds to the no-adrenergic receptor to activate the intracellular signalling cascade of G-protein coupling, which leads to an increase in intracellular cAMp concentration and activation of protein kinase A (PKA). Protein kinase A binds to RyRl via mAKAP, which forms a signal complex with skeletal muscle Ca2+ release (Reiken et al. 2003). Protein kinase A is acidified by RyRl to increase channel opening rate and sarcoplasmic reticulum Ca2+ release (Reiken et ai., 2003; Wehrens et al., 2004). 74 200815381 Data on skeletal muscle myofibrils have confirmed that intracellular Ca2+ leakage caused by increased RyR 1 activity after intense exercise is consistent with the maximal rate of sarcoplasmic reticulum Ca2+ release. Protein kinase A hyperphosphorylation of RyR 1 results in a lack of calstabinl (FKBP12) in the channel complex due to a decrease in calstabinl binding affinity. The long-term lack of calstabinl in the RyRl channel complex alleviates endogenous inhibition of the channel and causes uncontrolled intracellular Ca2+ leakage and reduced fatigue tolerance in the sustained high adrenaline state. Increased skeletal muscle fatigue in heart failure patients and heart failure animal models (Reiken et al. 5 (2003; Harrington et al., 1 997; Perreault et al., 1 993; Lunde et al. 2001; Lunde et Al. 1 99 8). In both patients with heart failure and animals, the skeletal muscle RyR1 channel isoform was found to be hyperphosphorylated by protein kinase A and lacks the stable calstabinl subunit (Reiken et al., 2003; Wehrens Et al., 2004). Fatigue improvement and RyRl over-linarization are accompanied by an increase in the Ca2+ spark wave frequency and a decrease in the Ca2+ spark wave amplitude in skeletal muscle fibrils of heart failure animals, which is consistent with intracellular Ca2+ leakage and muscle. Reduced Ca2+ concentration in the plasma network (Reiken et al., 2003). Therefore, muscle fatigue caused by long-term high adrenergic state is most likely to cause intracellular Ca2+ leakage via i RyRl channel defects (Reiken et al., 2003) However, it is not fully understood that the role of external Ca2+ ions in the contraction of mammalian skeletal muscle is not clear. The LTCC and RyR lines in the skeletal muscle and myocardium are different, and the skeletal muscle exhibits the LTCC ocls subunit. Tanabe et al. 5 1988) and RyRl (Marks et al., 1 989) and myocardium exhibit LTCC alc subunit (Mikami et al., 1989) and RyR2 (Nakai et al., 1990). Bone collateral 75 200815381 In muscle RyRl does not rely on Ca2+ influx through LTCC als to activate sarcoplasmic reticulum Ca2+ release, evidenced by persistent excitatory contraction coupling in skeletal muscle cells when externally removed or Ca2+ channel inhibitors are removed (Armstr〇ng et al. 5 1972; Dulhunty et al. 5 i 988; Gonzalez-Serratos et al., 1982). Subsequent experiments found that the activation system supporting RyRi is physically coupled to LTCC als (Ri〇s et al ·, 1987; Tanabe et al., 1 9 90). Ca2+ leakage from RyRl may increase the energy requirement of cells, compensating sarcoplasmic reticulum Ca2+ ATPase consumes more ATP, which may cause early skeletal muscle fatigue. Oxygen was measured directly on the muscle surface, and a significant increase in the overall content of ATp in the myoglobin Ca2+ ATPase in heart failure (Meyer et al., 1 998) was estimated to be due to intracellular Ca2+ leakage. Consistent with the decrease in sarcoplasmic reticulum ca2+ concentration due to Ca2+ leakage from RyRl, calstabinl, which specifically removes muscle, increases LTCC Ca2+ influx and reduces the maximum amount of Ca2+ release in voltage-gated cells (Tang et al., 2004) ). Recent studies have demonstrated functional deficits in the RyR1 channel in skeletal muscle during heart failure, similar to those found in the RyR2 channel in debilitated myocardium: protein kinase A over-acidification of RyR1 and lack of calstabinl (Marx et al., 2000; Reiken et al. , 2003; Wehrens et al., 2005). These findings confirm that defects in RyRl function alter intracellular Ca2+ treatment, resulting in early fatigue of skeletal muscle. The lack of calstabinl in the RyRl macromolecular complex also allows the channel to be uncoupled from the other channel and allows for the opposite of the gated control (Marx et al., 1998), thus providing compelling attention. Hypothesis to explain Ca 2 + cremation wave behavior in low-fatigue skeletal muscle (Ward et al., 2003). Therefore, the changes in RyRl can be reduced in weight and exercise tolerance in the skeletal muscle-specific power reduction mode of 76 200815381 (the machine is tired of the role of the body. The Latitude and the aerobic exercise can be fixed 4a ancient OI ^ Derogatory is a k咼 heart rate and a form of body movement that is done to improve performance. Aerobic; riding a bicycle and swimming. In some embodiments... 0 minutes aerobic swimming) requires twice a day for mice . Animals in the initial training course 3 days before the start of the swimming 2, 30 points twice; 2 days before the start, 45 points two

One day, 60 minutes, two 汝, called from A, the beginning of the day and the next are 9 〇 ^ Then the mice exercise twice a day for 2 or 21 days ρ swimming between classes... during the break, Keep mice warm and watery. Using a tunable mobile water tank, the mice were filled with water in an acrylic water tank (90 cm long; 45 cm wide; $knife) to a depth of 25 cm. Pumping creates a flow in the tank. The water flow rate at ί is maintained at a flow rate of 1 liter/min. Use a heater to hold the appropriate age and weight of the mice at 3 4 以 to rule out differences in physical strength. Forced swimming was used as an effective protocol to improve the aerobic exercise capacity of mice (Evangelista et al., 2003) to study the composition and phosphorylation status of skeletal complexes. Unexpectedly, after swimming for 90 weeks for 3 weeks, C57B16 wild sputum mice showed a significant increase in s phosphorylation (by protein kinase A), whereas CaMKII phosphorylation did not have oxygen to absorb running, I move (strongly adapt to the tour; open two On. And give exercise. ί 45 public swimming class water temperature fat fat skeletal muscle muscle channel divided into two RyR2 changes. 77 200815381

The RyR1 protein is stable, but the RyRl channel lacks the stable subunit calstabinl (FKBP 1 2). RyRi hyperphosphorylation and lack of calstabin 1 are consistent with leaky RyRi channels that cause intracellular Ca2+ leakage. To investigate the effect of duration of continuous exercise on RyRl Ca2+ release pathway defects, mice received swimming for up to 7 days, immediately after 7 or 21 days of sacrification. Acceptance of longer sustained exercise showed a significant increase in RyRl's protein kinase A hyperphosphorylation, which began on day 7 and reached saturation on day 21. In addition, studies have been conducted on mouse models of muscle wasting that are known to cause intracellular Ca2+ leakage and atrophic muscle changes. Surprisingly, the soleus muscle lacking dystrophin showed that it was

RyRl_Ser2844 lacks calstabinl without acidification. These results confirm the hyperphosphorylation of protein kinase A of RyRl-Ser2844 into a specific result of dysfunction of RyRl caused by exercise. At the same time, the characteristics of histological changes in the fast-twitch muscle of a 3-week exercise (swim) mouse were described. The cross-section of the mouse toe long muscle (M. exiewor heart/Mgw, EDL) showed histological changes consistent with myofibril regression caused by intracellular Ca2+ over-loading (caused by RyRl channel defects). Therefore, a continuous exercise of 9 minutes per day caused atrophy phenotype in the long toe muscle of normal C57B16 mice. Increased cAMP-dependent signaling pathway (Up-regUiati〇n) intracellular sarcoplasmic reticulum Ca2+ release increases excitatory contraction coupling when muscle tip performance is achieved. 200815381 (Reiken, 2003). Therefore, the skeletal muscle RyR1 Ca2+ release pathway transient protein kinase A acidification is an important mechanism for attack or escape response. Studies have confirmed that dysfunction of the RyRl Ca2+ release channel occurs during intense fatigue. Mice that received severe exercise for up to 3 weeks showed a significant increase in the degree of phosphorylation of protein kinase A by RyR 1, an increase in RyRi channel activity, and atrophic tissue changes. Long-term RyR1 hyperphosphorylation results in a lack of stable calstabinl, a functional deficit in the channel, suggesting intracellular Ca2+ leakage. At the muscle cell level, this pattern has recently been reported to increase the frequency of Ca2+ spark waves in mouse bone after muscle fatigue exercise (Wang et al., 2005). Furthermore, long-term intracellular Ca2+ leakage plays a role in malnutrition in mammalian skeletal muscle (Wang, 2005). This confirms the findings in previous cardiac muscles in which changes in hormones and tectonics contribute to intracellular Ca2+ leakage that triggers excitatory contraction coupling defects (Gomez, 1997). From these studies (Wehrens et al., 2005; Reiken et al., 2003) and 〇 & 2+ sparklet data in fatigue muscles (Fal & 1^61 & 1, 20 05), it can be inferred that intracellular sputum & 2 + leakage may be an important lesion that promotes muscle fatigue and causes muscle atrophic reorganization. Therefore, expanding the fatigue pattern to study the extent of in vivo, isolated muscle, muscle cells, granulocytes, and a single RyRl channel will characterize the detrimental effects of the tip on muscle fatigue and atrophic recombination. 1,4 - benzoquinone thiopurine derivatives prevent muscle dysfunction from intracellular Ca2+ leakage in the state of continuous activation of sympathetic nervous system (occurring in heart failure) (Wehrens et al., 2005). In some aspects, the present invention provides a RyCal compound and its use to increase skeletal muscle performance and reduce muscle dysfunction under sustained stress (as occurs in a fighting state). A therapeutic concept is rationalized by a defect in the RyRi Ca2+ release channel caused by aberrant regulation of the enzyme A: 200815381. The intracellular sarcoplasmic cytoplasm can be prevented by an agent that enhances the binding of the stable calstabul subunit to the RyRi channel complex. Net Ca2+ leakage and muscle dysfunction, and thus inhibition of sarcoplasmic reticulum Ca2+ leakage and muscle atrophic reorganization resulting from sustained pressure or exercise. The administration of agents to prevent sarcoplasmic reticulum Ca2+ leakage can thus enhance a number of important physiological abilities (these abilities are dysfunctional under stress). In some aspects, the invention provides for the use of a Rycai compound to increase the binding of caistabinl to RyR1 to prevent muscle fatigue. Using physiological (swim and treadmill running) and/or pharmacological (adrenalin: stimulant) to activate the sympathetic nervous system with animal models of varying degrees of exercise 'can test whether RyCal compounds can improve skeletal muscle function and prevent muscle Atrophic decline. This can lead to a pharmacological approach based on ectopic regulation of RyR 1 that results in improved human performance during sustained stress and prevents harmful tissue damage and thus reduces recovery time. In other aspects, the invention provides a pharmacological method based on ectopic regulation of RyRl, which is an improvement in human performance at sustained stress and prevents harmful tissue damage and thus reduces recovery time. \ · gj.R 1 胥 white kinase A deuteration: using two independent techniques (detection of RyRl protein kinase A phosphorus-containing epitope by specific antibody; by postphosphorylation test The radiolabeled phosphate was incorporated) to quantify the extent to which the skeletal muscle RyRl channel was phosphorylated by protein kinase A (PKA) for 2 days, 1 week, and 3 weeks. 80 200815381 Magnetic acidification: detection of the CaMKII phosphorus-containing epitope of RyR1 using a specific antibody developed by the inventor's laboratory to quantify the continuous movement of skeletal muscle RyRl channels for 2 days, 1 week and 3 weeks. The degree of phosphorylation of the protein (calmodulin) protein kinase II (CaMKII). This may characterize specific defects caused by long-term activation of protein kinase A phosphorylation and/or CaMKII signaling (caused by intracellular Ca2+ leakage mechanisms).

The lack of calstabinl in the RyRl complex: quantitation of the number of channel-stable subunits calstabinl (FKBP12) lacking on the RyRl channel complex for up to 2 days, 1 week and 3 weeks by immunoprecipitation. Phosphorylation of Ser2843 by protein kinase A caused the absence of calstabinl. Functional yRl Functional Defect - Development of a "leakage" RyRl channel in vivo: Current physiologic properties describing the activity and opening rate of RyRl single channel after 2 days, 1 week and 3 weeks of continuous exercise. This provides a comprehensive and sensitive assessment of the sarcoplasmic reticulum Ca2+ release channel defects and leakage mechanisms known to contribute to muscle fatigue. Furthermore, these data will develop a principle to prophylactically treat Ca2+ leakage of RyRl using a small number of RyCal lead compounds. Continuous exercise fatigue improvement quantifies fatigue in the living body using two independent athletic performance tests (swim and treadmill running). The treadmill test will be used in the animal subgroup with electrocardiogram telemetry to objectively assess the relationship between increased heart rate during exercise and fatigue symptoms. Furthermore, the catechin content of the blood and muscles will be determined to confirm the continued activation of the sympathetic nervous system. Three-week maximum fatigue swimming in i 81 200815381

The low flow rate of the ascending (bottom line state, which can prevent the small air from floating passively) causes progressive RyRl dysfunction and skeletal muscle atrophy. In order to quantify the fatigue time of the swimming material, the tracking system (San Diego Instruments Inc〇rporated), A automatically tracked and digitized the time_space of 8 mice. The degree of improvement in fatigue (defined as a 2d distance in time or a significant increase in activity) was assessed by the time of fatigue. In order to determine the maximum durability of exercise capacity (ie, the time to reach exhaustion), I wood exercise ability measured the longest swimming time at three flow rates of 7 liters/min. In order to reduce the intrinsic difference in swimming ability, mice with a maximum average swimming time variation exceeding the average swimming time of 4% or more will be excluded. When a mouse is unable to breathe out of the water, it is called fatigue and is saved at this point in time. To confirm systemic exhaustion after swimming, plasma lactate and pH values were determined in heparinized arterial samples. In order to objectively quantify the fatigue time during forced swimming, use video tracking system (vide〇 tracking syst(10)) (8 hearts)

Diego Instruments Incorporated), which automatically tracks and digitizes the 'space' of the 8 mice on the 2D plane. Silk Separation ^^ The function of the osseous muscle: continuous exercise with placebo or RyCal compound for up to 2 days, 1 week and 3 weeks, in vitro VZVC> describes muscle to fatigue stimulation or single-twitch The endogenous resistance of the shrinkage regimen. Two different forms of isolated skeletal muscle and long toe long muscle (exiewwr mountain g/Mrwm / owgws) will be tested to represent the fast-twisting muscle 82 200815381 and the soleus muscle represents the si〇w_twitch muscle. This test measures the effect of RyCal compounds on skeletal muscles in a state of fatigue or disease. Skeletal muscle contraction and diastolic system are importantly dependent on intracellular Ca2+ metabolism and RyR1 function and are therefore highly sensitive assays.星立离___冬骨路机中纤维中细漏漏和肌浆絪&2 + content: using the intracellular Ca2+ spark wave to evaluate the isolated skeletal muscle with flu〇_4 Ca2 + indicator Ca2+ leakage from the sarcoplasmic reticulum of the fibrils. The Ca2+ content of the sarcoplasmic reticulum was assessed using a caffeine pulse protocol that caused the complete release of the free Ca2+ group of the sarcoplasmic reticulum. Previous studies have confirmed that calcium leakage in myofibrils in mice (with heart failure) is caused by long-term high adrenaline status (Reiken 2003; Ward 2003; Gomez 2001; Cheng 1996) Ca2+ leakage from the sarcoplasmic reticulum Changes in mitochondrial integrity: The mitochondria in mouse skeletal muscle take Ca2+ under intense physiological muscle stimulation, resulting in sustained increase in granulocyte Ca2+ content and stimulation of mitochondrial metabolism (Rudolf et al ·, 2004) However, the myotubes of atrophic m-heart mice show a significant increase in Ca2+ intake (Robert V, Massimino ML, Tosello V, et al. Alteration in calcium handling at the subcellular level in mdx myotubes. J Biol Chem. Feb 16 2 001; 276(7): 464 7-465 1). Cytoplasmic over-loading of Ca2+ is a highly toxic event that is equivalent to the common pathway of cell death. Granulocyte plays an important role in cell death, and the spatial proximity of RyRl Ca2+ release to granulocyte Ca2+ captures 200815381 suggests that sarcoplasmic reticulum Ca2+ leakage during intense exercise can cause grain lines. Excessive Ca2+ loading, which has an effect on mitochondrial structure and function and may contribute to cell death. C a s p a s e -1 2 is located on the myopolynet, regulated by Ca2+ and involved in the apoptosis pathway induced by sarcoplasmic reticulum stress (Y〇ne da et al., 2001). In some aspects, the present invention describes the characteristics of fatigue movements on the membrane potential of mitochondria, which is studied by ingestion of rhodamine 123; mitochondrial expansion (representing mitochondria) Temporary permeability), studied by 520 nm spectrophotometer and optical scattering; Ca2+ content in the mitochondria was studied by culturing the cell with 45Ca2+ and then quantifying the radioactivity with a liquid scintillation counter; The mitochondria coactivating caspase enzyme measures cytochrome c; and the muscle preparation stains against TUNEL-positive nuclei. The in vivo effects of RyCal compound treatment on mitochondrial integrity and function will be assessed. Activation of intracellular proteases causes fragmentation of RyRl: a direct link between the level of Ca2+ and the proteolysis of intracellular substances in the cell is through the activation of Ca2+-dependent proteases, including mom activating proteases and caspase enzymes. . Calcium-activated protease activity is part of the mechanism of apoptosis. An increase in #5 activating protease activity has been observed in the mouse model of Duchenne muscular dystrophy (DMD), but the muscle-specific calcium-activating protease non-mutation mutation causes type 2A limb-type muscular dystrophy. (LGMD2A) (Tidball et al., 2000). These findings indicate that dysregulation of calcium-activated protease activity contributes to the development of muscle disease by disrupting normal regulatory mechanisms and systemic, non-specific increases in the ability to hydrolyze protein 84 200815381. The other components of the RyRl and Ca cycling mechanisms are caspase and the target of the activin enzyme and are cleaved by it (J〇hns〇n et al, ;

Shevchenko et al., 1 998). Therefore, the inventors will investigate whether the activity of Ca2+-dependent protease and/or caspase after intense exercise causes a cardiac rupture, and whether JTV519 can avoid the activation of non-specific proteolysis by inhibiting sarcoplasmic reticulum Ca2+ leakage. Histology of Skeletal Muscles in Labor Exercises #Continuing Exercise Atrophic changes can cause muscle fiber necrosis and progressive muscle weakness and weakness. Histological studies of skeletal muscle include the following analyses: eosinophilic over-contracted muscle fibers, necrotic fibers, ongoing muscle regeneration, and fibroblast proliferation in muscle tissue because muscle tissue is connected to connective tissue Substitution (fibrosis) is the main cause of permanent muscle weakness. Therefore, the inhibition of these changes by the Ry Cal compound can be tested. Traditional histological techniques can assess fiber size, split fiber, and variability in concentrated nuclei. Classification of mitochondria in skeletal muscle by total chemical analysis: Historically, the most widely used fiber type classification was based on mATPase (myofibrillar adenosine triphosphatase) activity, which was resolved by histochemical methods. Type I (low activity) and type II (high activity) fibers (Brooke et al., 1970). The Type II fiber is further classified into a Type IIA type and a ΠΒ type fiber by describing the characteristics of the pH instability of the mATPase. Additional fiber type characterizations for physiology, histochemistry, and ultra-microstructural methods show · Type I, medium-sized slow 85 200815381 oxidized fiber; Type IIA, red fast-reducing oxidized glycolytic fiber; and ΙΙΒ-type, White fast shrinking sugar fiber. Quantitative histochemistry can be used to determine oxidative and glycolytic abilities of mATP 83⁄4, succinate dehydrogenase and a-glycerophosphate dehydrogenase in sustained motility and/or RyCal compound therapy. Changes in the cross-sectional area of the myosin heavy chain (MHC) fiber. In order to determine the activity of myofibrillar ATPase, a cryosection using 1 μ μm thick cryopreservation (first culture in an acidic environment for 5 minutes or in an alkaline environment for 12 minutes) may be used. Succinate dehydrogenase staining will characterize the activity and differences between muscle fibers. Changes and improvements in oxidized muscle fiber types after myotonia treatment have previously been described in mice (Reininghaus et al., 1 988)

Classification of skeletal muscle fibers by immunocytochemical analysis: In order to quantify changes in the number and position of muscle nuclei and to distinguish between interstitial cell nuclei, the application of the nuclear to the nucleus (DAPI) to the basement membrane ( Fluorescence staining microscopy of laminin antibodies for morphometry. This technique is particularly important in determining the cross-sectional area of muscle fibers and the number of nuclei. Furthermore, this technique can be combined with the following affinity-purified antibodies for combined immunocytochemical analysis: C-terminal RyR2-5 029 antibody, RyRl-p2844 phosphorus-containing epitope-specific antibody (Wehrens et al ·, 2004), calstabinl (FKBP12) antibody (Jayaraman et al., 1992), myosin heavy chain isotype specific antibody (Rivero et al. 1 999) and α-actinin (Ruehr et Al·, 86 200815381 2 0 03), using a pre-established protocol with l〇μιη thick long toe muscles or low-temperature sections of soleus muscle (Moschella et al., 995). This method allows the inventors to classify myogens based on the myosin heavy chain content, metabolic activity, fiber size, and the degree of protein kinase A morphogenesis of RyR 1 in combination with cal stab ini during continuous exercise and/or Ry Cal compound treatment. Fiber and phenotypic changes. Furthermore, fiber type and immunocytochemical analysis data were linked to the following: isolated skeletal muscle function, general histological data, histochemical method, RyR 1 single channel function and mitochondrial data. Immunohistochemical staining was used to determine the binding of calstabin1 to RyRl and the protein kinase of RyRl after treatment with or without Ry Cal compound or Ry Cal compound in α-actinin positive compartment (Z-disc). A fiber type specificity of phosphorylation was improved, and an increase in the expression of #5 activating protease was measured in the myofibrillar region of the muscle fiber (Z disk; compartment containing RyRl channel). Skeletal muscle oxidizing capacity: Muscle oxidizing ability is related to the ability of skeletal muscle to use aerobic exercise. As a general experiment, the citric acid cycle and citrate synthase activity in muscle homogenate were evaluated by spectrophotometry to evaluate the activity of fast-shrinking skeletal muscle and slow-shrinking skeletal muscle oxidase. A similar protocol was used by the same research group to describe that the complex of coenzyme A and oxaloacetate is expected to increase its activity as measured (Evangelista et al., 2003). This test will be used to confirm the change in oxidative capacity observed in histochemical analysis. Improvement in plasma levels of creatine kinase (CK): plasma concentrations of creatine kinase (CK) and lactate dehydrogenase (LDH) can be determined (Santos et al., 2004; 87 200815381)

Thompson et al., 2004), as an indicator of muscle damage and inflammation after debilitating exercise, and used to determine the effects of RyCal compound therapy. Composition and function of RyR1 in cells: When stimulatory activation of B cells or T cell receptors, RYR1 in immune cells acts as a Ca2+ release channel (Sei t al., 2002; Kraev et al., 2003). For functional analysis, the peripheral mouse white blood cells were isolated by centrifugation of the mouse gold ball sample. White blood cells (1〇6/μl) were loaded with 1 μΜ fluo-3 of ethoxylated-mercapto esters (Molecular Pr〇bes, EUgene, l OR) by incubation at 25 ° for 30 minutes and tested in cells. The caffeine sensitivity of Ca2+ release. The properties of the components of the RyRi channel complex and protein kinase A phosphorylation will be described. Studying RyR1 in white blood cells can be used to monitor temporal changes in continuous exercise and RyCal compound therapy in vivo. Genetically modified mouse

The RyRl macromolecular signal complex plays an important role in regulating channel activation and coupling with excitatory contraction via the sympathetic nervous system. In the RyR 1 complex, mAKAP binds protein kinase A to phosphodiesterase PDE4D3 to the I channel and ligase pp 1 binds to the channel via the directional protein spinophilin. This signal module regulates the phosphorylation of protein kinase A on RyR2 Ser 2 8 43 as part of an “attack or escape” stress response. During normal exercise, two to three of the four Ser2843 protein kinase A phosphorylation sites in each tetrameric RyR1 channel are transiently phosphorylated via protein kinase A, resulting in an increase in RyRl channel activity. 88 200815381 Protein kinase A phosphorylation causes RyR1 kinase A phosphorylation to decrease the binding affinity of the stable protein calstabilU, resulting in a high Ca2+ dependence of the channel. The PDE4D3 color in the RyR1 macromolecular signal complex is protected against protein kinase A hyperphosphorylation and forms a negative feedback loop in the egg. Phosphorus in the RyRl complex rapidly reduces localized cAMP concentration and thus terminates channel activation. Accelerated muscle fatigue in mice lacking calstabinl in the RyRl complex can be tested. Therefore, both calstabinl and PDE4D3 can be regarded as "muscle dysfunction during excessive exercise or stress. Therefore, the extra component of the sub-complex can potentially be a novel drug for the treatment of harmful pharmaceutical agents to prevent the strong pressure molecules of the combatants. To prevent fatigue. Type 1 atrophic myotonia (D Μ 1) muscle-specific ugly MLR phenotype similar to atrophic muscle rigidity. Heavy and strong phenotypic sacral muscle regression and different repair degree signs, which is equivalent to The performance of a toxic long-form replicating variant of a short replicating variant. In addition to the sensitivity of the wild-type mouse swimming and treadmill running program to study muscle fatigue, the mice received the same motion-pressure protocol. Into the mouse. After the experiment is completed, the muscles are rapidly frozen in the nitrogen or further processed for stereoscopic vision to carefully cut a specific type of muscle activation system because the protein (FKBP12) plays a protective role in channel activation sensitivity. The role of acid diesterase in the activation of leukokinin A is protected by the lesser PDE4D3 or RyR2 complex of protein kinase A. RyRl scores the ground and/or the fatigue of the time. These single-sex mice are simulated, and the histopathology of the ugly SdLR muscle is less toxic than the less toxic. Sacrificial mice will be implanted in the subgroup of mice and tested in liquid histology. In the case of liquid nitrogen, the cold-cooled bundle was examined by histological analysis and immunohistochemical analysis. In all tissue samples, the degree of phosphorylation of protein kinase A in RyRl, the content of RyR2 macromolecular complexes (including caistabinl, protein kinase A, RII, mAKAP, ppi, spinophilin, PDE4D3, CaMKII) and the characteristics of a single channel will be examined. . The following characteristics were determined: activation and inhibition of Ca2+ sensitivity, Mg2+ inhibition, and changes in in vivo RyCal treatment. At the end of each single-channel experiment, the Ranol base was applied to the channel to confirm the identity of RyR. Mice will be able to test the beneficial effects of RyCal compounds in extreme genetic patterns of muscle fatigue and muscle wasting. The sensitivity of muscle fatigue can be studied using a swimming and treadmill running program. Implanted telemetry devices were implanted into mice two weeks before the mice received a exercise-pressure regimen. After the experiment is completed, the mice will be sacrificed and the muscles will be rapidly frozen in liquid nitrogen. After each experiment was completed, the muscles were cut and examined by histology and Western blotting. In all tissue samples, the degree of protein kinase A phosphorylation of RyRl, the content of RyR2 macromolecular complexes (including calstabinl, protein kinase A, RII, mAKAP, ppi, Spin〇philin, PDE4D3, CaMKII) and single channel will be examined. Characteristics. The following characteristics were measured: activation and inhibition of Ca2+ sensitivity, Mg2+ inhibition, and protein kinase A squaring. At the end of each single channel experiment, the linoreline was applied to the channel to confirm the identity of the RyR.

RyCal Compounds Prevent Muscle Fatigue and Muscle Depression In some aspects, the invention provides RyCal compounds that restore the normal function of hyperphosphorylated RyR1. Furthermore, applying PDE4D culling with 90

200815381 FKBP12 Single ligand deficiency (haploinsufficient) Whether J RyCal compounds prevent muscle fatigue. The above discussion establishes two animal models (mouse and rat) of muscle dysfunction and fatigue (cause of cause). This pattern of long-term activation of the sympathetic nervous system caused by exercise and stress is an important clue to critical defects in the empty Ca2+ release channel. Previous experiments in the heart failure mode that caused high levels of activation have been critical defects in RyRl that accelerate muscle fatigue. RyCal system for muscle fatigue during sustained exercise capacity, as well as research on fatigue-induced slow-shrinking skeletal muscle and fast-shrinking skeletal muscle function have been well established in rat and mouse heart failure patterns to determine categorical meat fatigue, so the inventors will Similar techniques were applied to test muscle fatigue after the immobilization protocol. During the continuous activation of the sympathetic nervous system (eg, but not the muscles that require maximal velocity, the long-term maximum pressure of intracellular sarcoplasmic reticulum Ca2 sympathetic nervous system is permanently activated by KyR 1 hyperphosphorylation and intracellular Ca2+ leakage. Internal Ca2+ leakage gradually causes muscle lesions, the specific duration of the myopathy and the significant decrease in maximum strength, and the additional ATP consumption of the sarcoplasmic reticulum (to compensate for uncontrolled leakage) Acceleration. The sarcoplasmic reticulum Ca2+ leakage is the direct cause of muscle fatigue inherent in myofibrils and is irreversible. Therefore, by calstabinl knot; the rat can be determined in continuous form to provide continuous ί to cause RyRl The sympathetic longitude is determined to contribute to the compounding effect of the compound in the mode of inhibition. Previously, the amine-induced muscle-sustaining animal is limited to fighting) dimension + circulation. Caused by possible • In the muscle, the cell sign for the superficial surface of the Ca2+ ATP sarcoplasmic reticulum Ca2+ is unique in the intense environment under the channel and 91 200815381 even under the Lili stable channel closed state to repair the tendon network Ca2+ leakage The agent (such as any RyCal compound), despite the constant stress, helps prevent fatigue from accelerating and promotes longer-term performance (Wehrens, 2000). This drug treatment is unique because of its toxic effects against major fatigue mechanisms and possibly preventing intracellular Ca2+ leakage. Furthermore, the molecular mechanism of this drug therapy is unique because its treatment contributes to specific defects in muscle fatigue, and since the mechanism of action of RyCal is to stabilize the normal RyFU channel by increasing the binding affinity of calstabinl, this is in contrast to blocking ions. The traditional method of channel function is significantly different. Recently, in the myofibrils and muscle wasting model after intense exercise (Wang et al., 2005), it was confirmed that the sarcoplasmic reticulum 2+ leakage may be caused by defects in the skeletal texture of the alkaloid receptor (RyR1). . Furthermore, long-term activation of the paternal nervous system (SNS) in heart failure causes the skeletal muscle (SM) in the body to fatigue due to the following reasons: · The RyRl complex lacks phosphodiesterase,

Serine 2844 protein kinase a over-scalage, lack of RyR1 complex

CalStaMnl and increased functional channel defects (Reiken et al., 2 03) RyRi dysfunction of the collateral muscle causes localized intracellular release and impaired global calcium current transients (Ward et al., 2 0 03). In the case of prolonged exercise, there is evidence that the change in macromolecular complexes is the lack of PDE4D3 in the RyRl complex, the hyperphosphorylation of protein kinase a in RyRl S 44, and the lack of cal stab in 1 in the RyR1 complex. From the secret, 1 Λ a rat model of repeated intense exercise in a time-dependent and active-dependent manner related to _ ' ^ 々 A ! After prolonged exercise, the RyRl macromolecular complex disk function is stabilized by these, biochemical changes in ▲, and the number is up to 92 200815381 days to weeks slowly recovering. Therefore, Ca2+ leakage of RyR1 limits the tip of the muscle and causes muscle damage during prolonged, intense exercise. The molecular mechanism of muscle fatigue has almost put the hypothesis that muscle fatigue is caused by lactic acid accumulation in the cytoplasm and potassium accumulation in the transverse tubules, and shifts attention to metabolism and mitochondrial regulation and signaling during prolonged exercise. Pathway research (Lin, Wu et al. 2002; Wu? Kanatous et al. 2002; Wang, Zhang et al. 2004). Although these alternative explanations are important, they are unlikely to directly explain the underlying abnormalities of excitatory contraction coupling found in fatigue muscles (Beirchtold, Brinkmeier et al. 2000). Here, the regulation of the skeletal muscle calcium release channel (RyR1) during long-term or high-intensity exercise is described. The recombination of the RyRl macromolecular complex during prolonged exercise is regulated by protein kinase a.

The over-phosphorylation of Ser2 8 44, the lack of ρ〇Ε4〇3, and the composition of caistabin' may play a role in determining muscle fatigue during prolonged exercise. Many positive effects of exercise-promoting organisms range from improving cardiovascular performance to taking two glucose intakes and normalizing fuel metabolism (G〇〇dyear and Kahn 199 8; Pollock, Franklin et al. 2). In heart failure, 'slight exercise training has been shown to improve skeletal muscle strength and reduce fatigue. The menstrual system adapts to more aerobic muscle characteristics (Minotti, Johnson et al. 1990; Lunde, Sjaastad et al. 2〇〇1; Meyer 2〇〇6). On the other hand, high-intensity exercises (such as those performed by marathon runners or long-distance bicycle riders) can cause significant muscle damage and damage to workers in a single event. 93 200815381 Performance for several days to weeks (O'Reilly, Warhol Et al. 1 987 ; Balnave and Thompson 1 993 , Komulainen and Vihko 1994). A mouse model of intense physiological movement is also described herein to examine changes in RyR 1 function and excitatory contraction coupling experienced by athlete elites, military personnel, or other individuals under intense stress. Daily swimming with a level treadmill running test can be constructed without excluding the physiological movements that include isometric or eccentric contractions of the hind limbs. Although the evidence of muscle injury in this type of exercise is less pronounced than the complete eccentric contraction (the data presented is easier to infer).

The biochemical changes identified in the RyRi macromolecular complex are consistent with the leakage-type calcium ion release channel. The single channel bilayer membrane data confirmed the leakage phenotype of the RyR1 channel (from the long-moving hind limb muscle), the number of resting calcium ions in the long-term exercise group (relative to the sedentary control group), the channel Has an increased turn-on rate. Reproduction of the RyR 1 complex biochemical changes in the two models of the mouse model, muscle specificity lacking calstabinl (call /) and lack of PDE4D3 (PDE4D-/-), can be found in motion defects. In another method, the absence of calstabinl is assessed by pharmacologically recombining the stabilizing subunits to RyR1 by the Ca2+ channel stabilizer Si〇7. Caistabin 1 caused by S107 recombined with RyR 1 resulted in an improvement in exercise capacity (measured by treadmill failure time), which was higher than that of mice lacking calstabinl by vehicle treatment in the same 2 day course. a//·/· 94 200815381 The lack of S107 effect in mice provides evidence of the molecular mechanism by which S107 does bind to RyRl via calstabinl. The in vitro fatigue protocol for intact isolated muscles is limited by the strength of the decline (which is mainly limited by tissue hypoxia) (zhang, Brut0I1 et al. 2006). Therefore, single-peg short muscle (Fdb) muscle fibers were isolated from sedentary and prolonged mice (with or without S 1〇7) to assess the reduction in tonic Ca 疲劳 during fatigue. The moving fibers with the recombined calstabin are relatively free of fatigue (Fig. 13). The effect of S1 07 does not appear to be due to the shift in characterization dynamics to the slower calcium ion cycle. S 107 corrects leakage in RyRl (from long-moving mice) as measured in the stationary phase of the lipid bilayer membrane for low levels of Ca2+. Ion spark waves do not occur frequently in all test conditions, which is consistent with most rare reports of spark waves in the skeletal muscle 'unless in a highly pathological state, such as hypoosmotic shock or muscular atrophy (Isaeva, Shkryl et al. 2005; Rios 2005; Wang, Weisleder et al. 2005), Ca2+ leakage caused by the high activity of the skeletal muscle receptors is not directly observed in the isolated muscle fibers. Many hypotheses show how changes in RyRl Ca2+ leakage cause muscle damage. However, these data do not reveal a muscle injury pathway that is solely responsible for the observed physiological outcome, which implies the effect of calcium-activated protease activation over prolonged and/or high-intensity exercise. Many teams have confirmed the primary mechanism by which the activation of protease-activated muscles causes muscle damage (Belcasu〇 1 993; Spencer and Mellgren 2002). As described herein, calcium-activated protease activity in the long toe muscles isolated 95 200815381 was increased after prolonged exercise (but reduced by S107 treatment), indicating that correction of the leaky RyRl prevents activation of calpain ( Figure 15). While other Ca2+-dependent pathways (such as caspase, gamma-promoting or calcitonin-dependent kinases) may contribute to the damage caused by leakage-type Arano receptors, the data of the present invention suggest a local increase in cytoplasmic Ca2+ The mechanism that causes harm. The hypothesis that leakage caused by Arano base receptors can cause muscle damage is further supported by evidence of reduced muscle damage in S 7 7 treated mice (as measured by serum creatine kinase) (Figure 15) . The data presented here show that changes in the RyRl macromolecular complex cause leakage phenotypes during prolonged, high-intensity exercise, which in turn impairs motor performance. Compounds In one aspect, the invention provides methods and uses comprising administering a compound of formula I:

Wherein, η is 0, 1 or 2; 96 200815381 q is 0, 1, 2, 3 or 4; each R is selected from the group consisting of: Η, halogen, -ΟΗ, -ΝΗ2, - Ν02, -CN, -CF3, -〇CF3 "N3, -S03H, -S( = 〇)2

Alkyl, -S(=0)alkyl, -0S(=0)2CF3, fluorenyl, -fluorenyl-alkyl, alkoxy, alkylamino, alkylarylamine, alkylthio , cycloalkyl, alkaryl, aryl, heteroaryl, heterocyclyl, heterocycloalkyl, alkenyl, alkynyl, (hetero) aryl, (hetero) arylthio and (hetero-) An arylamine group; wherein each fluorenyl group, -0-fluorenyl group, alkyl group, alkoxy group, alkylamino group, alkylarylamino group, alkylthio 'cycloalkyl group, alkylaryl group, aryl group can be optionally substituted , heteroaryl, heterocyclic, heterocycloalkyl, alkenyl, alkynyl, (hetero-)aryl, (hetero)arylthio and (hetero-)arylamine;

Ri is selected from the group consisting of H, keto, alkyl, alkenyl, aryl, alkyl, cycloalkyl, heteroaryl and heterocyclic; wherein each alkyl can be optionally substituted , alkene S, aryl, alkaryl, cycloalkyl, heteroaryl and heterocyclic; ~ R2 consists of the following Η, -C(=0)R5, -(CH2)m-Ri〇, alkane Selected from the group consisting of an alkyl group and a heterocycloalkylaryl group, a heteroaryl group: -CBuS)R6, -S〇2R7, a P(=〇)R8R9, an aryl group, an alkylaryl group a heteroaryl group, a cycloalkyl group, a ring; wherein each alkyl group, aryl group, cycloalkyl group, cycloalkylalkyl group and heterocyclic group are optionally substituted, and R3 is selected from the group consisting of the following: :H, -C02Y, -C( = 0)NHY, aryl, -O-SS, 户, 烯基, alkenyl, aryl, alkaryl, cycloalkyl, heterocyclyl and heterocycle a compound; an intermediate group, an alkyl group, an alkenyl group, a aryl group, an alkaryl group, a ring fluorene, a heteroaryl group and a heterocyclic group; and wherein the γ system is 97 200815381 Selected from the group consisting of hydrazine, alkyl, aryl, alkaryl, cycloalkyl a heteroaryl group and a heterocyclic group, wherein each alkyl group, aryl group, aryl group, cycloalkyl group, heteroaryl group and heterocyclic group are optionally substituted; R4 is selected from the group consisting of: Η , an alkyl group, an alkenyl group, an aryl group, an alkylaryl group, a cycloalkyl group, a heteroaryl group and a heterocyclic group; wherein each alkyl group, alkenyl group, aryl group, alkylaryl group, cycloalkyl group, hetero group can be optionally substituted An aryl group and a heterocyclic group; r5 is selected from the group consisting of: -NRi5Ri6, -(CH2)zNR"Ri6, -NHNR15R16, -NHOH, -〇r15, -C(-0)NHNR15r16, _c〇 2r15, _c(=o)nr15r16, _ch2x, aryl, alkyl, alkenyl, aryl, alkaryl, cycloalkyl, cycloalkylalkyl, heteroaryl, heterocyclyl and heterocycloalkyl; Wherein the thiol group, the alkyl group, the alkenyl group, the aryl group, the aryl group, the ring-based group, the cycloalkyl group, the heteroaryl group, the hetero-based group and the heterocyclic group are optionally substituted, and wherein the z-line 1 , 2, 3, 4, 5 or 6; R6 is selected from the group consisting of: R15, _nhnr15r16, -ΝΗΟΗ, _NR15R16, -CH2X, fluorenyl, alkenyl, alkyl, aryl, pyrene Aryl, cycloalkyl, cycloalkyl a pyridyl group, a heteroaryl group, a heterocyclic group and a heterocyclic group; wherein each of the fluorenyl, alkenyl, alkyl, aryl, aryl, cycloalkyl, cycloalkylalkyl, heteroaryl groups may be optionally substituted a group, a heterocyclic group and a heterocyclic group; R7 is selected from the group consisting of: 〇r15, -NR15r16, -NHNR15R16, -ΝΗΟΗ, -CH2X, alkyl, alkenyl, alkynyl, aryl An alkaryl group, a cycloalkyl group, a cycloalkylalkyl group, a heteroaryl group, a heterocyclic group and a heterocyclic group; wherein each alkyl group, alkenyl group, alkynyl group, aryl group, aryl group, Ring: 3⁄43⁄4 group, cycloalkyl group, heteroaryl group, heterocyclic group and heterocyclic group; 98 200815381 R8 and R9 are respectively composed of the following groups: a dilute group, an alkoxy group, a pyridyl group , an amine group, an aryl group, a hydrazine group, a heterocyclic group, a heterobasic group, and a heterocyclic ring to replace each fluorenyl group, alkenyl group, alkoxy group, alkyl group, aryl group, cycloalkyl group , cycloalkylalkyl, heteroaryl, heterocyclic

Rio is selected from the group consisting of OH, -SO2R11, -NHSC^Ru, c(= 〇)(r12), -0 00(1112) and -? ( = 0) 11131114; R 1 1 , R 1 2, R 1 3, and R 1 4 are respectively derived from the following: Η, OH, NH2, -NHNH2, -NHOH, sulfhydryl, alkyl, and burned a base group, a aryl group, a aryl group, a cycloalkylheteroaryl group, a heterocyclic group and a heterocycloalkyl group; wherein an alkenyl group, an alkoxy group, an alkyl group, an alkylamino group, an aryl group, a cyclyl group can be selected. The alkyl group, the heteroaryl group, the heterocyclic group and the heterocycloalkyl group X are selected from the group consisting of halogen, -C(=0)NR15R16, _NR15R16, 〇r15, _so2r7, and R15 and R16. The group consisting of the following group of neoyl, alkenyl, alkoxy, OH, NH2, alkyl, alkaryl, cycloalkyl, cycloalkylalkyl, heteroaryl, heterocyclic optionally substituted Each of a mercapto group, an alkenyl group, an alkoxy group, an aryl group, an alkylaryl group, a cycloalkyl group, a cycloalkylalkyl group, and a heterocycloalkyl group; and optionally, a 15 and R10 I are produced: 〇H, brewed a aryl group, an aryl group, a cycloalkyl group, a selective amine group, an aryl group, an alkyl group and a heterocycloalkyl group; :-NR15R16, , nhc = 〇(r12), a selected group, a dilute group , oxygenated •, cycloalkylalkyl, • substituted each a thiol group, a aryl group, a calcyl group, -CN, -C02R15, and -p(=o)r8r9; - selected from the group consisting of hydrazine, hydrazino, aryl, alkyl and heterocycloalkyl; The base, the amine amine 'heteroaryl, the heterocyclic ring may form a heterocyclic ring with the 99 200815381 nitrogen to which it is combined, and the heterocyclic ring may have a substituent; the nitrogen in the benzothiazepine ring may be optionally a four Grade nitrogen; and its mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, water complexes, solvates, complexes and prodrugs. In certain embodiments of Formula I, if q is 〇 and n is 〇, then R2 is not Η, Et, ·€( = 0)ΝΗ2, (= 〇)NHPh, -C( = S)NH - n-Butyl, -C( = 0)NHC( = 0)CH2C1, -C( = 0)H, -C( = 0)Me ' -C( = 〇)Et, -C( = 0)CH = CH2, -S( = 〇)2Me or -S( = 0)2Et; and if q is 0 and η is 1 or 2, then R2 is not -C( = 0)Me, C( = 0 ) Et, -S( = 0)2Me or _S( = 0)2Et; in addition, if q is 1 and R is methyl, chloro or fluorine at position 6 of benzothiazepine, then R2 It is not Η, Me, -C( = 0)H, -C( = 0)Me, -C( = 〇)Et, -C( = 0)Ph, -S( = 0)2Me or -S( = 0) 2Et ; and further, if q is 1, n is 〇 and r is OCT3, thiol or Ci-Cs alkoxy at position 7 of the stupid and sulphur ring, then R2 is not Η, - c(=〇)ch=ch2 or

In one embodiment, the invention provides methods and uses comprising the application of a compound of formula 1 (as described above] but the compound is not S24 or S68. In one embodiment, the invention provides methods and uses comprising the administration of a compound of formula ha: 100 200815381

Where: η is 0, 1 or 2, q is 0, 1, 2, 3 or 4; each R is selected from the group consisting of: Η, halogen, -OH, -ΝΗ2, _n〇2 , -CN, -CF3, -〇Cf3, N3, _s〇3H, s(=〇)2 alkyl, -s(=〇)alkyl, -〇s(=o)2CF3, fluorenyl, alkyl, Alkoxy, alkylamino, alkylthio, cycloalkyl, aryl, heterocyclyl, heterocycloalkyl, alkenyl, alkynyl, (hetero)aryl, (hetero-)arylthio with a hetero-)arylamino group; which may be substituted or unsubstituted with each fluorenyl group, alkyl group, alkoxy group, amphoteric group, thiol group, cycloalkyl group, aryl group, heterocyclic group, heterocycloalkyl group, alkenyl group , alkynyl, (hetero-)aryl, (hetero-)arylthio and (hetero-)arylamine; r2 is selected from the group consisting of: Η, —C = 0(R5), - C = S(R6), -s〇2R7, -P( = 〇)R8R9, -(CH2)m-Ri〇, alkyl, aryl, heteroaryl, cycloalkyl, cycloalkylalkyl and hetero a cyclic group; wherein each alkyl group, aryl group, heteroaryl group, cycloalkyl group, cycloalkylalkyl group and heterocyclic group may be substituted or unsubstituted; R5 is selected from the group consisting of: NRl5Rl6, 10 1 200815381 -(CH2)zNRi 5R16 , -NHNRi 5R16 , -NHOH , -OR15 , -c(=o)nhnr15r16, -C〇2r15, -C(=0)NR15R16, -CH2X, sulfhydryl, alkyl, hydrazine Alkyl, alkynyl, aryl, cyclopropenyl, cycloalkyl, heterocyclyl and heterocycloalkyl; wherein may be substituted or unsubstituted each fluorenyl, alkyl, alkenyl, alkynyl, aryl, ring An alkyl group, a cycloalkylalkyl group, a heterocyclic group and a heterocyclic group, wherein z is 1, 2, 3, 4, 5 or 6; R6 is selected from the group consisting of: -OR", -NHNR15R16, -NHOH, -NRi5R16, -CH2X, decyl, alkenyl, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl and heterocycloalkyl; wherein each may be substituted or unsubstituted Amidino, alkenyl, alkyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl and heterocycloalkyl; R7 is selected from the group consisting of Η, 〇r15, -NRi5Ri6, -NHNR15R16, -NHOH, -ch2x, alkyl, methoxy, alkynyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl and heterocycloalkyl; wherein each may be substituted or unsubstituted Alkyl, alkenyl, alkynyl, aryl, cycloalkyl, Alkyl group alkyl, heterocyclyl and heterocycloalkyl;

The Rs and R9 systems are respectively selected from the group consisting of 〇Η, fluorenyl, alkenyl, alkoxy, alkyl, alkylamino, aryl, cycloalkyl, cycloalkylalkyl, a heterocyclic group and a heterologous alkyl group, which may or may not be substituted for each fluorenyl group, aryl group, alkoxy group, alkyl group, acryl group, aryl group, cycloalkyl group, cycloalkylalkyl group, heterocyclic group and Heterocycloalkyl; R10 is selected from the group consisting of -Nr15r16, OH, -NHS02R", -C(= 〇)r12, _NH(C = 〇)Rl2, 102 200815381 -0 (C = 0) Ri2 and -P〇0)r13r14; M system 0, 1, 2, 3 or 4;

Rn, R!2, Ru and R!4 are respectively selected from the group consisting of the following: Η, OH, NH2, -NHNH2, -NHOH, thiol, alkenyl, alkoxy, alkyl An alkylamino group, an aryl group, a cycloalkyl group, a cycloalkylalkyl group, a heterocyclic group and a heterocycloalkyl group; wherein the fluorenyl group, the alkenyl group, the alkoxy group, the alkyl group, the acryl group may be substituted or unsubstituted. , aryl, cycloalkyl, cycloalkyl, heterocyclyl and heterocycloalkyl; X is selected from the group consisting of halogen, -CN, -CO2R15, -C( = 0)NR15Ri6 , -NR15R16, -OR15, -S02R7 and -P( = 0)R8R9 ;

Rl 5 and Rl6 are respectively selected from the group consisting of H, decyl, alkenyl, alkoxy, OH, hydrazine 2, alkyl, alkylamino, aryl, cycloalkyl, cycloalkyl An alkyl group, a heterocyclic group and a heterocycloalkyl group; wherein each of a decyl group, an alkenyl group, an alkoxy group, an alkyl group, an alkylamino group, an aryl group, a cycloalkyl group, a cycloalkylalkyl group, or a heterocyclic group may be substituted or unsubstituted. a cyclic group and a heterocycloalkyl group; and, optionally, r15 and R16 may form a heterocyclic ring together with the nitrogen to which they are bonded, the heterocyclic ring may have a substituent; the nitrogen in the benzothiazepine ring may be optionally It is a four-stage nitrogen; and its mirror image isomer, non-image isomer, tautomer, pharmaceutically acceptable salt, hydrate, solvate, complex and prodrug. In one embodiment, 'if q is 0 and η is 0, then R2 is not Η, Et, 〇( = 0) ΝΗ 2, ( = 〇) NHPh, _C( = S)NH-n-butyl, 103 200815381 _C( = 0)NHC( = 0)CH2C1, -C(,H,_c( = 〇)Me, -c(,Et, -C( = 〇)CH = CH2, -S( = 0)2Me Or _s( = 〇)2Et; and if q is 0 and n is 丨 or 2, then R2 is not -C( = 0)Me, -C( = 0)Et, -S 〇)2Me Or s( = 〇)2Et; in addition, if q is 1 and R is methyl, gas or fluorine at position 6 of benzothiazepine, then R2 is not H, Me, _CK = C)&gt ;H, = -C( = 0)Et, -C( = 0)Ph, -S( = 〇)2Me or _s(==0)2Et ;

Furthermore, if q is 1, n is 0 and R is the position of benzothiazepine ring No. 7

When OCT3, hydroxy or C1-C3 alkoxy, then R2 is not H -c(=o)ch=ch2 or

In certain embodiments, the present invention provides methods and uses comprising a compound of the formula wherein each R line is selected from the group consisting of: hydrazine, halogen, -OH, 〇Me, -NH2, -N02 , -CN, -CF3, -〇CF3, -Ν3, -SbOhCrC* alkyl, 4( = 0)(^-0:4 alkyl, -S-C1-C4 alkyl, -OS( = 〇)2CF3 , Ph, -NHCH2Ph, •C(=〇)Me, -0C(=0)Me, morpholinyl and propyl phenyl; and n is 0, 1 or 2. In other embodiments, the invention Methods and uses are provided comprising the administration of a compound of formula Ia, wherein R2 is selected from the group consisting of: -C = 〇(R5), -C = S(Rg), -SO2R7, a P(=0)R8R9 And -(CH2)m-R10. 104 200815381 In yet another embodiment, the invention provides methods and uses comprising administering a compound of formula Ib:

Wherein R' and R" are respectively selected from the group consisting of hydrazine, halogen, -OH, -NH2, -N〇2, -CN, -CF3, -OCF3, -N3, -SO3H, - S(=0)2 alkyl, -S(=0)alkyl, -0S(=0)2CF3, fluorenyl, alkyl, alkoxy, alkylamino, alkylthio, cycloalkyl, aryl a heterocyclic group, a heterocycloalkyl group, an alkenyl group, an alkynyl group, a (hetero-)aryl group, a (hetero-)arylthio group and a (hetero-)arylamino group; wherein the fluorenyl group or the alkane may be substituted or unsubstituted Alkyl, alkoxy, alkylamino, cycloalkyl, aryl, heterocyclyl, heterocycloalkyl, alkenyl, alkynyl, (hetero-)aryl and (hetero)arylthio; R2 and η Generally as defined in the above formula Ia; and its mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes and prodrugs. In certain embodiments, the invention provides methods and uses comprising administering a compound of Formula Ib, wherein the R' and R" systems are each selected from the group consisting of: hydrazine, halogen, -OH, OMe, -NH2 , -N〇2, -CN, 105 200815381 CF3, -OCF3, -N3, -S pOhCrCU alkyl, -8 (= 0)(^-(^4 alkyl, -S-Ci-CU alkyl, -0S( = 0)2CF3, Ph, -NHCH2Ph, -C( = 0)Me, · 0 (Ι: ( = 0) Μ 6, 淋 基 and propylene groups; and η is 0, 1, or 3. In some instances, R' is Η or OMe and R" is Η. In other embodiments, The invention provides methods and uses comprising the administration of a compound of formula Ib, wherein R2 is selected from the group consisting of: -c=o(r5), -C=S(R6), -SO2R7, -p(=o) R8r9 and -(CH2)m-R10. In yet another embodiment, the invention provides methods and uses comprising administering a compound of formula Ic:

Wherein each R, R7, q and η are as defined in the above formula Ia; and as mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvents Compounds, complexes and prodrugs. In certain embodiments, the present invention provides methods and uses comprising the administration of a compound of Formula Ic, wherein each R is selected from the group consisting of: 106, 2008, 381, respectively, halogen, -OH, OMe, _NH2 -CF3, -OCF3, -N3, -S( = 0)2Ci-C4 alkyl, sulfhydryl, -S-C1-C4 alkyl, -OS( = 〇)2CF3, Ph -C( = 0)Me, - 0C (= 0)Me, morphinyl and propenyl; or 2 〇 In other embodiments, the invention provides methods and uses including administration wherein R7 consists of #-OH, -NR15R16, alkyl, A aryl group, an aryl group, a cycloalkylheterocyclyl group and a heterocycloalkyl group; embodiments in which a aryl group, a cycloalkyl group, a cycloalkylalkyl group, a heterocyclic group and a heterocyclic group may be substituted or unsubstituted are further examples. In the present invention, the method and use of the inclusion complex are provided: -N〇2, -CN, SfCOCrC^ alkane, -NHCH2Ph, and n-system 0,1 丨Ic compound Ic is selected from the group consisting of: cycloalkylalkyl , alkyl, alkenyl, alkyl. Application I-d

Wherein R, and R" are respectively composed of the group consisting of -OH, -NH2, -N〇2, -CN, -CF3, _〇Cf3 s(=o)2 alkyl, -s( =〇)alkyl, -os(=〇)2CF3, selected from: oxime, halo, -Ν3, -S03H, fluorenyl, alkyl, 107 200815381 alkoxy, alkylamino, alkylthio, cycloalkyl , aryl, heterocyclic, heterocycloalkyl, alkenyl, alkynyl, (hetero-)aryl, (hetero-)arylthio and (hetero)arylamine; and wherein each may be substituted or unsubstituted Mercapto, alkyl, alkoxy, amphetamine, cycloalkyl, aryl, heterocyclyl, heterocycloalkyl, alkenyl, alkynyl, (hetero) aryl, (hetero-) arylthio R7 and η are as defined in the compounds of the above formula a; and mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, Complexes and Prodrugs. In certain embodiments, the invention provides methods and uses comprising a compound of the formula wherein R' and R" are each selected from the group consisting of: hydrazine, halogen, -OH , OMe, -ΝΗ2, CN, -CF3, -OCF3, -Ν3, -S( = 0)2Ci-C4 alkyl, = alkyl, -S - C1 - C 4 alkyl, -0 S (= Ο) 2 CF 3, ph, - n HC Η 2 P h, -C( = 0) Me, -0C (= 0) Me, morphine and propyl group; and n is 〇, 1 or 3. In certain instances, R' is hydrazine or OMe and R" is η. In other embodiments, the invention provides methods and uses comprising a compound of the formula wherein R? is selected from the group consisting of: - OH, -NRi5Ri6, alkyl, alkenyl, aryl 'cycloalkyl, cycloalkylalkyl, heterocyclic and heterocycloalkyl; wherein each alkyl, alkenyl, aryl, ring may be substituted or unsubstituted An alkyl group, a cycloalkyl group, a heterocyclic group and a heterocyclic group. In one embodiment, the invention provides methods and uses comprising the administration of a compound of formula Ie: 108 200815381

Wherein each R, Rs, q and n are as defined in the compound of formula k above; and mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, Solvates, complexes and prodrugs. In certain embodiments, the present invention provides methods and uses comprising a compound of the formula wherein each R line is selected from the group consisting of: hydrazine, halogen, -OH, 〇Me, -ΝΗ2, -N02 , -CN, -CF3, -OCF3, -Ν3, -SpOhCi-CU alkyl, -S( = 0)Ci-C4 alkyl, -S-Ci-C4 alkyl, -OS( = 〇)2CF3, Ph , -NHCH2Ph, -C(=0)Me, -0C(=0)Me, morphinyl and propenyl; and n is 0, 1 or 2, in other embodiments, the invention provides for the administration of a compound of formula Ie The method and use, wherein R5 is selected from the group consisting of -NR15R16, -(CDzNRuRu, NHOH, -OR15, -CH2X, alkyl, alkenyl, aryl, cycloalkyl, cycloalkyl An alkyl group, a heterocyclic group and a heterocycloalkyl group; wherein each of a fluorenyl group, an alkyl group, an alkenyl group, an aryl group, a cycloalkyl group, a cycloalkylalkyl group, a heterocyclic group and a heterocyclic group can be substituted or unsubstituted. 109 200815381 In certain embodiments, the invention provides methods and uses comprising administering a compound of Formula Ie, wherein R5 is at least one calibration group (such as fluorescent, biologically luminescent, chemically luminescent, colored, and irradiated) Substituted alkyl group. The base of the cursor is selected from the following: dioxodecane boron difluoride compound (bodipy), dansui, luciferin, rhodamine, dezhou red, cyanine Dyes, tar brain, coumarin, Cascade BlueTM, Pacific Blue, Molina Blue, Olegang Green, 4', 6-diamidino-2-phenylindole (DAPI), Debbie Dyes, fluorescein, propidium iodide, pyrrole, arginine, and variants and derivatives thereof. In another embodiment, the invention provides methods and uses comprising the administration of a compound of formula If:

(If) where R' and R" are respectively selected from the group consisting of Η, halogen, -0Η, -ΝΗ2, ·Ν〇2, -CN, -CF3, -0CF3, -N3, - S03H, •s(=o)2 alkyl, -s(=o)alkyl, -os(=o)2cf3, fluorenyl, alkyl, alkoxy, alkylamino, alkylthio, cycloalkyl , aryl, heterocyclic, heterocyclic 110 200815381 alkyl, alkenyl, alkynyl, (hetero-)aryl, (heteroarylthio and (hetero-)arylamino); and which may be substituted or unsubstituted Each mercapto group, alkyl group, alkoxy group, alkylamino group, cycloalkyl group, aryl group, heterocyclic group, heterocycloalkyl group, alkenyl group, alkynyl group, (hetero-)aryl group, (hetero-) aryl sulfide R5 and η are as defined in the above formula Ia; and as mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes And certain prodrugs. In certain embodiments, the invention provides methods and uses comprising the administration of a compound of the formula If the R' and R" systems are each selected from the group consisting of: H v halogen, - OH ' OMe, -NH2, -N02, -CN, -CF3, -OCF3 -N3, alkyl, alkyl, -S-Ci-C4 alkyl, -〇S( = 0)2CF3, Ph, -NHCH2Ph, -C( = 0)Me, -0C( = 0)Me, morpholine And propylene; and n is 〇, j or 3. In some embodiments, R' is Η or OMe and R" is Η. In other embodiments, the invention provides methods and uses comprising administering a compound of formula If, Wherein Rs is selected from the group consisting of -NR15R16, -(CH2)ZNR15R16, -NHOH, -OR15, -CH2X, alkyl, alkenyl, aryl, cycloalkyl, cycloalkylalkyl, a heterocyclic group and a heteroalkyl group; wherein each of a fluorenyl group, an alkyl group, an alkenyl group, an aryl group, a cycloalkyl group, a cycloalkylalkyl group, a heterocyclic group and a heterocycloalkyl group may be substituted or unsubstituted. In an embodiment, the invention provides methods and uses comprising administering a compound of formula Ig: 111 200815381

Nr15r16 α-g)

Constructs, tautomers, pharmaceutically acceptable salts, non-mirromeric hetero hydrates, solvates, complexes and prodrugs. In certain embodiments, the present invention provides methods and uses comprising a compound of the formula wherein each R-line is selected from the group consisting of: hydrazine, halogen, -OH, 〇Me, -NH2, _CN , -CF3, -OCF3, ·Ν3, alkyl, -s( = 〇)Cl_C4 alkyl, -S-Ci-C4 alkyl, -〇S( = 0)2CF3, Ph, -NHCH2Ph, -C( = 0) Me, -0C (= 0)Me, morpholinyl and propenyl; and n is 〇, i or 2 〇 In other embodiments, the invention provides methods and uses comprising administering a compound of formula Ig, wherein Rl 5 And Rl6 are respectively selected from the group consisting of hydrazine, OH, NH2, alkyl, alkylamino, aryl, cycloalkyl, cycloalkylalkyl, heterocyclic and heterocycloalkyl; Wherein each alkyl group, alkylamino group, aryl group, cycloalkyl group, cycloalkylalkyl group, heterocyclic group and heterocyclic group 112 200815381 alkyl group may be substituted; and optionally, Ri 5 and R 16 may be bonded thereto. N together form a heterocyclic ring which can be substituted. In certain embodiments, the invention provides methods and uses comprising administering a compound of Formula I-g, wherein W is oxime or S. In yet another embodiment, the invention provides methods and uses comprising the administration of a compound of formula I-h:

Wherein R' and R" are respectively selected from the group consisting of Η, halogen, -ΟΗ, -ΝΗ2, -Ν〇2, -CN, -CF3, -OCF3, -N3, -S03H, - S(=0)2 alkyl, -s(=0)alkyl, -0S(=0)2CF3, decyl, alkyl, alkoxy, alkylamino, alkylthio, cycloalkyl, aryl a heterocyclic group, a heterocycloalkyl group, an alkenyl group, an alkynyl group, a (hetero-)aryl group, a (hetero-)arylthio group and a (hetero-)arylamino group; wherein the fluorenyl group or the alkane may be substituted or unsubstituted Alkyl, alkoxy, alkylamino, cycloalkyl, aryl, heterocyclyl, heterocycloalkyl, alkenyl, alkynyl, (hetero-)aryl 113 200815381, (hetero-)arylthio; R15 , R16 and 11 are as defined in the compound of the above formula 1-&; and their mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates , Complexes and Prodrugs. In certain embodiments, the present invention provides methods and uses comprising the administration of Formula Ih, wherein R' and R" are respectively selected from the group consisting of hydrazine, halogen, -OH, OMe, NH2, -N〇2, -CN -CF3, -OC F3, -N3, -8( = 0)2(^-(34 alkyl, -8( = 0)(^-(^4 base, -S-Ci-C4 alkyl, -0S( = 0)2CF3 , Ph, -NHCH2Ph -C( = 0)Me, -0C( = 0)Me, morpholinyl and propenyl; and n is 0, or 3. In some instances, R' is Η or OMe and R" In other embodiments, the invention provides methods and uses comprising the administration of a compound of Formula Ih, wherein the Ri 5 and Ri6 lines are each selected from the group consisting of hydrazine, OH, NH 2 , alkyl, alkylamine a group, an aryl group, a cyclic group, a cycloalkylalkyl group, a heterocyclic group and a heterocycloalkyl group; wherein each group, an alkylamino group, an aryl group, a cycloalkyl group, a cycloalkylalkyl group, a heterocyclic group Heteroalkyl; and optionally, Ri5 and R16 may form a heterocyclic ring with the nitrogen to which they are bound, and the heterocyclic ring may be substituted. In certain embodiments, the invention provides a method comprising administering a compound of formula Ih. And use, wherein W is 0 or S. In a further embodiment, the invention provides a method and use comprising the application of a compound of formula I: alkane ring physicochemicals of a mixture of alkane 114 200815381

Wherein Rl7 is selected from the group consisting of: ^^ nu16, -NHNR15R16, -ΝΗΟΗ, one OR15, -CH2X, alkenyl, real 1

a crater, a cycloalkyl group, a cycloalkylalkyl group, a heterocyclic group and a heterocycloalkyl group; wherein the group can be substituted, the group is not substituted, and the aryl group, the aryl group, the cycloalkyl group, the cycloalkyl group, and the heterocyclic soil are substituted. Each R, q and n of the heterocyclic ring is as defined in the compound of the above formula Ia - generally; and its mirror image isomer, non-image isomer, tautomer, ~ ^, and is acceptable for learning Salts, hydrates, solvates, complexes and prodrugs. In certain embodiments, the present invention provides methods and uses comprising the application of a formula of a τ, 丨" compound, wherein each R is selected from the group of J Γ / τ, respectively: Η, halogen, -OH ' OMe, -NH2, N〇2, _CN, -CF3, _〇CF3, _n3, _s(==〇)2CVC4 alkyl, ·8( = 〇)(:ι_ε4 alkyl, -S-Ci -CU calcination, -〇S( = 〇)2CF3, Ph, -NHCH2Ph, -C( = 〇)Me, -〇C( = 0)Me, morphinyl and propenyl; and n is 0, 1 or 2 In other embodiments, the invention provides methods and uses comprising the administration of a compound of formula Ii, wherein R17 is -NR15R16 and -or 15. Some other solids 115 200815381 -NHEt, -NHPh, -NH2, in the case of 'R1 7 - Ο -, - Ο M e, - NE t, or -NHCH 2 aridinyl. In one embodiment, the invention provides methods and uses comprising a compound of the formula:

Wherein R' and R" are respectively selected from the group consisting of: hydrazine, halogen-OCF3, -N3, -SO3H, -, -OH, -NH2, -N02, -CN, -CF3, -S ( = 0) 2 alkyl, -s( = o)alkyl, -〇S( = 0)2CF3, fluorenyl, alkyl, alkoxy, alkylamino, alkylthio, cycloalkyl, aryl a heterocyclic group, a heterocycloalkyl group, an alkenyl group, an alkynyl group, a (hetero-)aryl group, a (hetero-)arylthio group and a (hetero-)arylamino group; and wherein the thiol group may be substituted or unsubstituted, Alkyl, alkoxy, alkylamino, cycloalkyl, aryl, heterocyclic, heterocycloalkyl, alkenyl, alkynyl, (hetero-)aryl, (hetero-)arylthio; R17 Selected from the group consisting of: -NRl5Rl6, -NHNRi5Ri6, -NHOH, -〇R15, -CH2X' alkenyl, aryl, cyclohexyl, cycloalkylalkyl, heterocyclyl and heterocycloalkyl Which may or may not be substituted for each of the fluorenyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl and heterocycloalkyl groups of the group 200818381; n is as defined in the compound of the above formula ϊ-a And its mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable Classes, hydrates, solvates, complexes and prodrugs. In certain embodiments, the present invention provides methods and uses comprising the administration of a compound of Formula Ij, wherein the R, and R" systems are respectively composed of the following Selected from the group: Η, halogen, -OH, OMe, -NH2, -N〇2, -CN, -CF3, -〇CF3, _N3, -StOhC^CU alkyl, -S( = 0)Ci-C4 alkane Base, thiol, -OS(=0)2CF3, Ph, -NHCH2Ph, -C(= 〇)Me, -0C(=0)Me, morpholinyl and propenyl; and n is 〇, 1 or 3. In certain instances, R' is Η or 〇 Me and R" is Η. In other embodiments, the invention provides methods and uses comprising administering a compound of Formula Ij, wherein R17 is -NR1SR16 or 〇r15. Certain other In an embodiment, Ri7 is -OH, -OMe, -NEt, -NHEt, -NHPh, -NH2 or -NHCH2 pyridyl. In another embodiment, the invention provides methods and uses comprising administering a compound of formula Ik: 117 200815381

Wherein R' and R" are respectively selected from the group consisting of: Η, halogen, -ΟΗ, -ΝΗ2, -N〇2, -CN, one cP3, -OCF3, -N3, -S03H, - s(=o)2 alkyl, -s(=〇)alkyl, -0S(=0)2CF3, fluorenyl, alkyl, alkoxy 'alkylamino, alkylthio, cycloalkyl, aryl a heterocyclic group, a heterocycloalkyl group, an alkenyl group, an alkynyl group, a (hetero-)aryl group, a (hetero-)arylthio group and a (hetero-)arylamino group; and wherein each of the aryl groups may be substituted or unsubstituted, An alkyl group, an alkoxy group, an acryl group, a cycloalkyl group, an aryl group, a heterocyclic group, a heterocycloalkyl group, an alkenyl group, an alkynyl group, a (hetero-)aryl group, a (hetero-) arylthio group;

Rl8 is selected from the group consisting of -NRl5Rl6, -c(=o)nr15r16, _(c = 〇)〇R15, a 0R15, an alkyl group, an aryl group, a cycloalkyl group, a heterocyclic group, and a a calibration group; wherein each alkyl group, aryl group, cycloalkyl group and heterocyclic group may be substituted or unsubstituted; wherein p is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; 0, 1 or 2; and its mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes and prodrugs. In certain embodiments, the invention provides methods and uses comprising a compound of the formula wherein R' and R" are selected from the group consisting of 118, 2008, 15, 381, respectively: hydrazine, halogen, -OH, OMe, - NH2, -N02, -CN, -CF3, -OCF3, -N3, -S(=0)2Ci-C4 alkyl, -S(= 〇)Ci-C4 alkyl, -S-CrC* alkyl, - 0S( = 0)2CF3, Ph, -NHCH2Ph, -C(=0)Me, -0C(=0)Me, morpholinyl and propenyl; and n is 0, 1 or 3. In some instances, R 'System or OMe and R' system. In other embodiments, the invention provides methods and uses comprising administering a compound of formula Lk, wherein Ru is selected from the group consisting of: NRi 5R16, -(C = 0) 0Ri 5, an ORi 5, hospital a base, an aryl group, and a calibration group; and wherein each alkyl group and aryl group may be substituted or unsubstituted. In some instances, m is 1 and Ris is Ph, C( = 〇)〇Me, C( = 0)〇H, aminoalkyl, NH2, NHOH or NHCbz. In other examples, m is hydrazine and Rl8 is a C1-C4 alkyl group (such as decyl, ethyl, propyl and butyl). In still other examples, m is 2 and R丨s is p 洛 洛 (P y r r 〇 1J d i n e ), piperididine, piperazine or morpholine. In some embodiments, m is 3, 4, 5, 6, 7, or 8 and Ris is a cursor-based base selected from the group consisting of dipyrrolidine boron difluoride compound, dansyl , luciferin, rhodamine, texas red, phthalocyanine dye, tar brain, coumarin, casque blue, pacific blue, maolina blue, olere green, 4, , 6 - dimercapto -2 - Benzoin bee (DAPI), Debbie dye, Fluorescent yellow, Cellulose, Pyridine, arginine, and the above variants and derivatives. In yet another embodiment, the invention provides methods and uses comprising the administration of a compound of formula 1-1: 119 200815381

J

(1-1) wherein R' and R" are respectively selected from the group consisting of H, halogen, -OH, -NH2, -N〇2, -CN, -CF3, -〇CF3, - N3, -S03H, -S(=0)2 alkyl, -S(=0)alkyl, -0S(=0)2CF3, fluorenyl, alkyl, alkoxy, alkylamino, alkylthio, Cycloalkyl, aryl, heterocyclyl, heterocycloalkyl, alkenyl, alkynyl, (heteroaryl, (hetero-)arylthio and (hetero-)arylamino); Each mercapto group, alkyl group, alkoxy group, alkylamino group, cycloalkyl group, aryl group, heterocyclic group, heterocyclic compound group, alkenyl group, alkynyl group, (hetero-)aryl group, (hetero-) aryl sulfide R6 and η are as defined in the compound of the above formula a; and mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, gluten Compounds, Complexes, and Prodrugs. In certain embodiments, the invention provides methods and uses comprising the administration of a compound of Formula 1-1, wherein R' and R" are each selected from the group consisting of: Η, halogen, -0H, OMe, -NH2, -N02, -CN, CF3, -OCF3, -N3, -S( = 0) 2Cl-C4 alkyl, 4 (= 0) (^-04 alkyl, -S-Ci-C4 alkyl, -08...0)]???!!, -:^!^^! ?!!, 120 200815381 -C( = 0)Me, -0C( = 0)Me, morpholinyl and propenyl; and ηι or 3. In some instances, R is Η or 〇Me and R" is Η In other embodiments, the invention provides methods and uses comprising an application formula wherein R6 is selected from the group consisting of -NR15R16, -NHNR15R16, _R15, -NHOH, -CH2X, alkenyl, alkyl , aryl, cycloalkyl, cycloalkylalkyl, heterocyclylalkyl; wherein may be substituted or unsubstituted with each fluorenyl, alkenyl, alkyl, cycloalkyl, cycloalkylalkyl, heterocyclic group and Heterocycloalkyl. Some of the real R6 systems are -NR15Ri6 (such as -NHPh, pyrrolidine, hexahydropyridine, hydrazine, orolin, etc.). In some other examples, the r6 is a calcined oxygen such as a 0_tBu 〇 in a further In the embodiments, the present invention provides methods and uses comprising administering an analog compound: g 0, 1 compound selected: fluorenyl, and heterocyclic aryl, in the case, hexahydroindenyl, exemplified

Wherein R, and R" are respectively selected from the group consisting of -OH, -NH2, -N〇2, -CN'-CF3, -〇cf3, -N3, :Η, halogen S03H, 121 200815381 -S( = 0)2 alkyl, -s( = 〇)alkyl, -OS( = 〇) 2cf3, decyl, alkyl, alkoxy, alkylamino, alkylthio, cycloalkyl , aryl, heterocyclic, heterocycloalkyl, alkenyl, alkynyl, (heteroaryl, (hetero-)arylthio and (hetero-)arylamino); and wherein each thiol group may be substituted or unsubstituted , alkyl, alkoxy, alkylamino, cycloalkyl, aryl, heterocyclyl, heterocycloalkyl, decyl, alkynyl, (hetero) aryl, (hetero-) arylthio; R8 , R9 and η are as defined in the compound of the above formula Ia; and mirror image isomers, non-image isomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes And certain prodrugs. In certain embodiments, the invention provides methods and uses comprising the administration of a compound of Formula Im, wherein R' and R" are each selected from the group consisting of: hydrazine, halogen, -OH , 〇Me,, -cf3, -ocf3, -n3, -s(= 2Cl-C4 alkyl...s(=〇)Ci C4 alkyl, -S-Ci.C* alkyl,_0S(=0)2CF3, Ph, _NHCH2Ph, -C( = 〇)Me, _〇C( = 0) Me, morpholinyl and propenyl; and n is 〇, ! or 3. In some instances, R, is Η or 〇Me and R" is η. In other embodiments, the invention provides for administration. A method and use of a compound of Formula Im, wherein R8 and R9 are each independently alkyl, aryl, 〇H, alkoxy or alkylamine. In certain instances, r8 is Ci-C4 alkyl (such as methyl, Ethyl, propyl and butyl); and R9 is aryl, for example phenyl. In other embodiments, the invention provides methods and uses comprising a compound of the formula: 122 0, 0, 200815381

S- where:

Rd is CH2 or NRa;

Ra is H, -(CVC6 alkyl)-aryl, wherein the aryl is a disubstituted phenyl or benzo[13]dioxo-5-yl group or an amine protecting group (eg , B 〇c group);

Rb is a hydrogen of an alkoxy group (for example, a decyloxy group). Exemplary compounds of Formulas I-n include, but are not limited to, S1〇i, S102, S103, S114. In certain other embodiments, the present invention provides compounds of Formula I-o.

among them:

Re-alkyl)-phenyl, -(CVC6 alkyl)-C(0)Rb, or substituted or unsubstituted -C^Cg alkyl; and Rb-OH or mono-(Ci- C6 alkyl), and 123 200815381 wherein the phenyl or substituted alkyl group may be substituted with one or more of the following groups: halogen, hydrocarbyl, -CrQ alkyl, -0-(Cl-c6 alkyl), -NH2, -NH (Ci-C6 alkyl), -N (Ci-C6 alkyl) 2, cyano or dioxolane Exemplary compounds of the formula I - 包括 include, but are not limited to, S 1 0 7, S 1 1 0, SI 11, S120 and S121 〇 In certain other embodiments, the invention provides a compound of the formula Ι-p:

S

I-P where: R. - (CrCs alkyl)-nh2, alkyl)-0Rf, wherein Η or —(:(0)_((:1<6) alkyl or one (Ci_c6 alkyl) 4HRg, wherein R is a benzene Carboxybenzyl. Exemplary compounds of Formula Ip include, but are not limited to, S109, S122, S123. In a non-limiting example, Formula la, lb, Ie, If,

Ag Ah, In is represented by compounds SI 01, S102, and SI 03. In a non-limiting example, Formula 1a, lb, le, If, nu are represented by the compound sl4. In a limiting example, the formula la, Ib, 1 is represented by SI 07. In a non-limiting example of non-1, the formulas Ia, Ib, Ie, If are represented by S108. Right-wing non-restriction 124 200815381 In the sexual example, the formulas I a, I b, I e, I f, I p are represented by S 1 0 9 . In a non-limiting example, the formulas la, lb, Ik, Io are represented by S 1 1 0 . In a non-limiting example, the formula la, lb, Ik, Ιο is represented by S 1 1 1 . In a non-limiting example, the formula la, lb, Ic, Id is represented by S 1 1 2 . In a non-limiting example, the formula la, lb is represented by S 1 1 3 . In a non-limiting example, the formula la, lb, Ie, If, Ig, Ih is represented by S1 14. In a non-limiting example, the formula la, lb, Ig, Ih, II is represented by S 1 15 . In a non-limiting example, the formula la, lb, Ig, Ih is represented by S 1 16 . In a non-limiting example, the formula la, lb, Ie, If is represented by S 1 17 . In a non-limiting example, the formula la, lb, Ie, If is represented by S 1 18 . In a non-limiting example, the formula la, lb is represented by S 1 1 9 . In a non-limiting example, the formula la, lb, Ik, 1 is represented by S120. In a non-limiting example, the formulas la, lb, Ik, Io, Ip are represented by S121. In a non-limiting example, the formulas la, lb, Ie, If, Ip are represented by S122. In a non-limiting example, the formulas la, lb, Ie, If, Ip are represented by S123. Formula I, Ia, Ib, Ic, Id, Ie, If, Ig, Ih, Ii, Ij, Ik, 1- 1. Im, In, Io and Ip Compounds of formula II are useful in the treatment or prevention of abnormalities and diseases associated with the RyR receptor. Examples of the above compounds include, but are not limited to, S 1 , S2, S3, S4, S5, S6, S7, S9, S11, S12, S13, S14, S19, S20, S22, S23, S24, S25 as defined herein. , S26, S27, S36, S37, S38, S40, S43, S44, S45, S46, S47, S48, 125 200815381 S49, S50, S51, S52, S53, S54, S55, S56, S57, S58, S59, S60 , S61, S62, S63, S64, S66, S67, S68, S69 > S70, S71 'S72, S73, S74 > S75 > S76, S77, S78, S79, S80, S81, S82, S83, S84, S85, S86, S87, S88, S89, S90, S91, S92, S93, S94, S95, S96^S97' S98, S99, S100 > S101, S102, S103, S104, S105, S107, S108, S109, S110 S111, SI 12 'SI 13 > S114, SI 15 > S116, S117, S118, SI 19 > S120, S 1 2 1 , S 1 22 and S 1 23 . In certain embodiments, the compound is isolated and substantially pure. In a certain embodiment of the method, the compound is not S4. In another embodiment, the compound is not S7. In another embodiment, the compound is not S8. In another embodiment, the compound is not S 1 0. In another embodiment, the compound is not S20. In another embodiment, the compound is not S24. In another embodiment, the compound is not S25. In another embodiment, the compound is not S26. In another embodiment, the compound is not S27. In another embodiment, the compound is not S36. In another embodiment, the compound is not JTV-519. Certain RyCal compounds of the invention have the following construction: S1 126 200815381

127 200815381

128 200815381

Q

Ch2i S7

S9

O

129 200815381 s.

S12 N

NH

S13

\ S14

S19 130 200815381

S23 131 200815381

S24 S-24

\

O

S25 132 200815381

S36

133 200815381

S40 s S43 134 200815381

S45

135 200815381 〇·

NH〇 S46

S48

136 200815381

S49

137 200815381 S52 〇·

Ο

S33 Ν

S54 Ο

138 200815381

139 200815381

S60 140 200815381

S a Γν

ΝΗ〇 S61

S ΝΗ 2 S62

S- is

S63 141 200815381

S64 K S66

OH S S67

142 200815381

S70

S71 143 200815381

S72

S73

144 200815381 S75

S76 〇 S77

145 200815381

S78

146 200815381 Ο

S81

147 200815381

OCMe3 S85

S86 200815381 S87

OCMe3

S89 149 200815381

S90

S91 〇

S92 150 200815381

PhCH2NH

χ N OCMe3 S93

〇CMe3 S94 S95

OCMe3 151 200815381

〇 OC OCMe3

02N S97

152 200815381

S99 again 〇CMe3 S100 N3 s-

153 200815381

S102

▽ S S103

154 200815381

S107

〇

MeO S108

A^NHCbz 〇

155 200815381 S110

Sill

S112

o IIS-OH II 〇

156 200815381

S115

Ο

A^^^^Bodipy 157 200815381 ο-

S119

S120

S121

S122 158 200815381

ο S123 In one embodiment of the invention, with respect to the compound of formula I, C = 0 (R5) or S02R7, then R is located at positions 2 and 3 on the phenyl ring. In another embodiment of the invention, wherein the compound of formula I is C = 0 (R5) or S02R7, then each R is selected from the group consisting of Η, halogen, -OH, -NH2, respectively. -N〇2, -CN-S03H, fluorenyl, alkyl, alkylamino, cycloalkyl, heterocyclyl, benzyl, alkenyl, (hetero-)aryl, (hetero-)arylthio and (hetero -) arylamino; fluorenyl, alkyl, alkoxy, alkylamino, cycloalkyl, heterocyclylalkyl, alkenyl, (hetero-)aryl, (hetero)arylthio with a hetero-)arylamino group or a plurality of groups selected from the group consisting of: N, Ο, -S-, -CN, -N3, -SH, nitro, keto, fluorenyl An alkoxy group, an alkylamino group, an alkenyl group, an aryl group, a (hetero-)cycloalkyl ring group. In another embodiment of the invention, wherein the compound of formula I is C = 0 (R5) or S02R7, then at least two R groups are attached. Further, at least two R groups are bonded to the benzene ring, and both groups are bonded to the 2, 3 or 5 position on the benzene ring. Further, if R2 or No. 5, if R2 is formed, -N3, a heterocyclic ring is burned, and a heterocyclic ring may be used in one generation: a halogen group, a calcination and a (hetero-) group, and if R2 is a benzene ring, R groups are R. 159 200815381 is otherwise selected from the group consisting of hydrazine, halogen, -OH, -NH2, _M〇2, _n3, _s〇3H, fluorenyl, alkyl, alkylamino, cycloalkyl , heterocyclyl", heterocycloalkyl, alkenyl, (hetero-)aryl, (hetero-)arylthio and (hetero-)arylamine; wherein each thiol, alkyl, alkoxy, An alkylamino group, a cycloalkyl group, a heterocyclic group, a heterocycloalkyl group, an alkenyl group, a (hetero-)aryl group, a (heteroquinone arylthio group and a (hetero-)arylamine group may be used one or more of the following respectively Substituted groups of selected groups are substituted: halogen, N, oxime, -S-, -CN, -N3, -SH, nitro, keto, decyl, alkyl, alkoxy, alkylamino, Alkenyl, aryl, (hetero, -)cycloalkyl and (hetero-)cyclo. t · In another embodiment of the invention, with respect to the compound of formula I, if r2 is c=o(r5), Then r5 is selected from the following group: -NRi6, _(CH2)zNRi5Ri6, NHNHR16, NHOH, -ORi5, CONH2NHRi6, CONR16, ch2x, fluorenyl, aryl, cycloalkyl, cycloalkylalkyl, heterocyclyl and heterocycloalkyl; wherein each thiol, aryl, cycloalkyl, naphthenic The alkyl group, heterocyclic group and heterocycloalkyl group may be substituted with one or more groups selected from the group consisting of halogen, N, fluorene, -s-, -CN, -N3, nitro a keto group, a decyl group, an alkyl group, an alkoxy group, an alkylamine v group, a fluorenyl group, an aryl group, a (hetero-)cycloalkyl group and a (hetero-) ring group. In another embodiment, the invention provides Use of the compound of formula II: 160 200815381

Wherein R = OR', SR', NR', alkyl or halide and R, == alkyl, aryl or hydrazine, and wherein R may be at position 2, 3, 4 or 5. Formula II is also described in detail in the co-pending application Serial No. 10/680,988, the disclosure of which is incorporated herein in its entirety by reference. The lightness of the activity The compound of the present invention reduces the opening ratio of RyR by increasing the affinity of FKBP12 (calstabinl) and FKBP12.6 (calstabin2) for each of protein kinase a phosphorylated RyR and RyR2. In addition, the compound normalizes the gating ability of the mutant RyR channel (including the mutant RyR2 channel associated with CPVT) by increasing the binding affinity of FKBP12 (calstabinl) to FKBP12.6 (calstabin2). Therefore, the compounds of the present invention prevent abnormalities and symptoms involved in the regulation of RyR receptors (especially, the ruler 7111 and the ruler 7 are 2 receptors). Examples of such abnormalities and symptoms include, but are not limited to, cardiac abnormalities and diseases, skeletal muscle abnormalities and diseases, cognitive abnormalities and diseases, malignant fever syndrome, diabetes, and sudden infant death. Cardiac abnormalities and diseases include (but are not limited to) irregular heartbeat abnormalities and diseases, irregular heartbeat abnormalities and diseases caused by exercise, sudden cardiac death, sudden cardiac death caused by exercise, congestive heart failure, and chronic obstructive pulmonary disease hypertension. Irregular heartbeat abnormalities and diseases including irregular heartbeat abnormalities and diseases caused by exercise, its package 161 200815381

(but not limited to) atrial and ventricular arrhythmia, atrial fibrillation and ventricular fibrillation; atrial frequency arrhythmia and ventricular pacing arrhythmia; atrial frequency and ventricular frequency; catecholamine polymorphic heart to frequency ( CPVT) is a variant caused by its movement. Skeletal muscle abnormalities and diseases include, but are not limited to, skeletal muscle fatigue, exercise-induced skeletal muscle fatigue, muscular dystrophy, bladder dysfunction, and incontinence. Cognitive disorders and diseases include, but are not limited to, Alzheimer's disease, various types of memory loss, and age-dependent memory loss. The compounds of the present invention treat these abnormalities and symptoms by increasing the binding affinity of FKBP12 (calstabinl)-RyRl to increase the binding affinity of FKBP12.6 (calstabin2)-RyR2. In accordance with the above, the present invention provides a method of limiting or avoiding a decrease in the amount of FKBP (calstabin) bound to RyR in an individual cell. Here "RyR" includes RyRl, RyR2 and RyR3. In addition, FKBP includes both FKBP12 (calstabinl) and FKBP12.6 (calstabin2). Therefore, "FKBP bound to RyR" means FKBP 12 (calstabinl) which binds to RyR1, FKBP12.6 (calstabin2) which binds to RyR2, and FKBP12 (calstabinl) which binds to RyR3. Here, "RyR" also includes "RyR protein". And "RyR analogues". The "RyR analog" is a functional variant of RyR protein having RyR biological activity, which is 60% or more homologous to the amino acid sequence of the RyR protein. The RyR of the present invention may be un-acidified, acidified (e.g., by protein stimulating A) or hyperphosphorylated (e.g., by protein A). Here, the term "RyR biological activity" is used in the analysis status of 162 200815381, which means the protein or Shengyuetai activity that physically binds or binds to ca 1 stabi ' ' when RyR is RyRl and RyR3, calstabin is FKBP12 (calstabinl And 'calstabin is FKBP12.6 (calstabin2) when RyR is RyR2 (ie, nearly twice or five times more binding than the negative control group). Here, "FKBP" includes both "FKBP protein" and "FKBP analog", regardless of whether FKBP is FKBP12 (calstabinl) or FKBP12.6 (calstabin2). Unless otherwise indicated herein, a "protein" shall include a protein, a protein domain, a polypeptide or a peptide, and any such fragments. "FKBP analog" is a functional variant of FKBP protein with FKBP biological activity, whether f KBP is F κ BP 1 2 (calstabinl) or FKBP12.6 (calstabin2), its homology to the FKBP protein amino acid sequence 60% or higher. The term "r ρ Κ BP biological activity" as used herein, in the context of the analysis herein, refers to a protein or peptide activity that physically binds or binds to unphosphorylated RyR2 or RyR2 that is not hyperphosphorylated (ie, near The background binding force of the negative control group was twice or five times higher than the binding strength). FKBP binds to the RyR channel and each RyR subunit is relative to a ρΚΒΡ molecule. Thus, the term "FKBP conjugated to RyR" herein includes a FKBP12 (calstabinl) protein molecule that binds to a RyR1 protein subunit or an FKBP12 tetramer that binds to a RyRl tetramer; binds to a RyR2 protein subunit a FKBP12.6 (calstabin2) protein molecule, or a FKBP12.6 tetramer that binds to a RyR2 tetramer; a FKB P12 (cal stab ini) protein molecule that binds to a RyR3 163 200815381 protein subunit, or a combination FKBP12 tetramer of RyR3 tetramer. Therefore, "FKBP combined with RyR" means "fkbpi2 combined with RyRi", "FKBP12.6 combined with RyR2", and "FKBP12 combined with RyR3". According to the method of the present invention, Fkbp which binds to RyR in an individual cell contains a "reduction" or "abnormality" of $, meaning that the amount of FKBP bound to RyR in an individual cell is detectably reduced, decreased or reduced. When such a reduction is restricted, hindered, prevented, hindered or reduced in any way by administration of a compound of the invention, the above-described reduction in the cells of the individual can be limited or prevented such that the FKBP content of the intracellular binding to RyR in the individual is higher than that of the other. Examples of compounds are administered. Standard analysis and techniques can be used to determine the FKBP content of an individual that binds to RyR. These techniques include simple assays in the prior art (eg, 'immunity techniques, heterozygous assays, immunoprecipitation, Western blot analysis, fluorescent imaging techniques and / or radiation detection, etc.), as well as any of the analysis and detection methods disclosed herein. For example, proteins are isolated and purified from individual cells by standard methods well known in the art, including, but not limited to, essential cell extraction (eg, using a soluble protein intercalating agent); Affinity purification; chromatographic techniques such as flow thin layer chromatography (FTLC) and high performance liquid chromatography (HPLC); immunoprecipitation (using antibodies); and precipitation (eg, using isopropanol with, for example, "Trizol Reagents). The isolation and purification of the protein is followed by electrophoretic analysis (e.g., on a sodium sulfosyl sulfate "SDS-polyacrylamide" gel). The reduction in FKBP content bound to RyR in an individual can be determined by administering a compound of formula I according to the invention (in accordance with the method described below) to a FKBP content that binds to RyR after a suitable period of time, as compared to 164 200815381; Or the above limitation or prevention. The reduction of FKBP content bound to RyR in an individual cell can be restricted or prevented by, for example, inhibiting the isolation of FKBP from RyR in an individual cell; increasing the binding between FKBP and RyR in an individual cell; or stabilizing intracellular cells in an individual. RyR-FKBP complex. In addition, the reduction of FKBP content in the intracellular binding of RyR in an individual can be limited or prevented by: directly reducing the acidified RyR content in the individual cells; or indirectly reducing the intracellular acidified RyR content, for example, to regulate Or an enzyme that regulates the phosphorylation of RyR function or content (eg, protein kinase A) or another endogenous molecule. In one embodiment, the method of the invention reduces at least 10% of the phosphorylated RyR content in the cells. In another embodiment, at least a 20% phosphorylated RyR content is reduced. Individuals of the invention may be in vitro and in vivo systems, including, without limitation, isolated or cultured cells or tissues; non-cell WWo assay systems; and animals (eg, two-story, birds) , fish, mammals, marsupials, humans, pets (such as tracing, dogs, monkeys, horses, mice or rats) or economic animals (such as cows or pigs). Individual cells include striated muscle cells. The striated muscle is a muscle type in which the repetitive unit of contractile myofibrils (myofibril) is neatly arranged throughout the cell, resulting in lateral or diagonal stripes observed under an optical microscope. Examples of striated muscle cells include, but are not limited to, random (skeletal) muscle cells and cardiomyocytes. In one embodiment, the cell line used in the methods of the invention is a human cardiomyocyte. The term "cardiomyocyte" as used herein includes, for example, myocardial fibers found in the heart muscle. The myocardial fibrosis consists of a series of continuous cardiac muscle cells or cardiomyocytes connected end to end in the intervertebral disc (i n t e r c a 1 a t e d d i s k s ). These intervertebral discs have two types of cell junctions: expanded desmosomes that extend along their direction of detection and gap junctions that extend along their longitudinal portion. By administering a compound of the invention to an individual, a reduction in the FKBP content of the RyR bound to the somatic cell can be limited or prevented; this also applies to contact between the individual cell and the compound of the invention. The compounds of the invention are calcium ion channel modulators. In addition to modulating Ca2+ concentration in cardiomyocytes, the compounds of the invention also modulate, for example, Na+ ion flux and inward-rectifier K+ ion flux in guinea pig ventricular cells, as well as inhibition of, for example, intraventricular atrial cell delay. R+ (deUyed_rectifia) K+ ion current. Pharmaceutical Compositions The compounds of the present invention are formulated into pharmaceutical compositions suitable for in vivo administration in a biocompatible form for administration to human subjects. According to another embodiment, the present invention provides a pharmaceutical composition comprising a compound of the present invention in admixture with a pharmaceutically acceptable diluent and/or agent. The pharmaceutically acceptable carrier must be "acceptable" in the phase of the combination with the ingredients and is not deleterious. The pharmaceutically acceptable carrier used herein is selected from the group consisting of pharmaceutically acceptable carriers. Each of 166 200815381 organic or inorganic substances' includes analgesics, buffers, binders, disintegrating agents, diluents, emulsifiers, excipients, extenders, slip agents, solubilizers, stabilizers, suspending agents , tonicity agent, carrier and augmentation agent. Pharmaceutical additives such as fragrances, colorants, flavors, preservatives, and sweeteners may also be added as needed. Examples of acceptable pharmaceutical carriers include carboxymethyl cellulose, microcrystalline cellulose, glycerin, acacia, lactose, magnesium stearate, methylcellulose, powder, saline, Sodium alginate, sucrose, starch, talc, water, and the like. The pharmaceutical formulations of the present invention are prepared by methods well known in the pharmaceutical art. For example, a compound or a diluent of the compound of the present invention is combined with a carrier and/or a diluent to form a suspension or solution. Alternatively, one or more accessory ingredients such as buffers, flavors, surfactants and the like may also be added. The choice of carrier will depend on the solubility and chemical nature of the compound, the route of administration, and standard pharmaceutical practice. A compound of the invention is administered to an individual by contacting the compound with a subject cell (e.g., a cardiomyocyte) within a body. The present invention is contacted (e.g., introduced) into individual cells using a technique such as introduction and transfer of proteins, nucleic acids, and W drugs. Examples of methods of contacting a cell with a compound of the invention (ie, treating the cell) include, without limitation, absorption, electroporation, infiltration, immersion, liposome delivery, Transfection methods, transfusi〇n, vectors, and other drug delivery vehicles and methods. When the target cell is located in a particular part of the individual, the compound of the invention is preferably delivered directly to the cell by injection or other means (e.g., 167 200815381 to introduce the compound into blood or other body fluids). Detection of target cells in individual tissues by standard detection methods in the prior art, examples of such detection include (not to be): immunological techniques (eg, immunohistochemical staining), fluorescent imaging techniques, and display Microtechnology. In addition, the compounds of the invention can be administered to a human or animal subject by conventional procedures. This step includes, without limitation, oral, sublingual or buccal, parenteral, transdermal, inhalation or intranasal, intravaginal, intrarectal. And intramuscular administration. Parenteral administration of a compound of the invention includes by epithelial, intracapsular, intracranial, intradermal, intrathecal, intramuscular, intraocular, intraperitoneal, intraspinal, intrasternal, vascular Injection into the inside, vein, stromal tissue, subcutaneous or sublingual, or by catheter. In one embodiment, the drug is administered to the individual by delivery to the muscle of the individual (including, but not limited to, the myocardium of the animal). In one embodiment, the music is administered to the individual via drug delivery to the cardiomyocytes via a catheter inserted into the individual's heart. In other embodiments, the agent is administered via subcutaneous pumping. Formulations for oral administration of the compounds of the invention may be in the form of capsules, lozenges, powders, granules, suspensions or solutions. The formulation contains a common additive such as lactose, mannitol, corn starch or potato starch. The formulation also contains a binder such as microcrystalline cellulose, cellulose derivatives, acacia, corn starch or gel. In addition, the formulation also contains a disintegrating agent such as corn starch, potato starch or sodium carboxymethylcellulose. The formulation also contains anhydrous disc acid hydrogenation (dibasic calcium phosphate 168 200815381, the most chloramphenic acid 瓿 瓿 瓿 脏 ) ) ) ) ) ) ) )) or sodium starch glycolate. Thereafter, the formulation may contain a lubricant such as talc or magnesium stearate. For parenteral administration (ie, a drug introduced via a route other than the digestive tract), the compound of the present invention is incorporated into a sterile aqueous solution that is isotonic with the blood of the individual. Such a formulation is prepared by dissolving the solid active ingredient in a solution containing, for example, sodium. An aqueous solution prepared from a physiologically compatible substance such as glycerin and having pH-reducing water compatible with physiological conditions, and then sterilizing the solution. The formulation can be placed in a single or multiple dose container, such as a sealed amoules or vial. The party can be delivered by any mode of injection, including (not limited to) extramuscular, intracapsular, intracranial, intradermal, intrathecal intramuscular, intraocular, intraperitoneal, intraspinal, intrasternal, intravascular, Intravenous, stromal tissue, subcutaneous or sublingual injection, or by means of a catheter inserted into the heart of an individual. For transdermal administration, the compound of the present invention is conjugated to a skin penetration aid such as propylene glycol, polyethylene glycol, isopropanol, ethanol, oleic acid, i-methylpyrrolidone or the like to enhance the compound of the present invention. The permeability of the skin and allows the compound to penetrate the skin into the blood. The composition/permeabilizer composition of the present invention is further combined with a polymer such as ethyl cellulose, hydroxy cellulose, vinyl acetate, polyvinyl pyrrolidone, etc., to make the composition And allowing the gelled composition to be dissolved in a solvent such as dichloromethane, evaporated to the desired viscosity and then applied to the backing material to form a patch 169 200815381. In certain embodiments, the composition It is made into a single-dose, lozenge, capsule or single-dose vial. Depending on the desired clinical effect, a unit dose, i.e., a therapeutically effective dose, will be administered during a clinical trial designed to treat each disease of the selected compound. The present invention also provides an article suitable for treating a body abnormality (e.g., cardiac abnormality). The compounds of the present invention, or one or more of the compounds of the invention as described herein, comprise a drug analog of the mother knife λ. The article is co-packaged with instructions for indicating the condition of treatment, combination, and/or prevention of the pharmaceutical composition. For example, the article contains a unit dosed human that is capable of treating or preventing a muscle disease as disclosed herein, and indicates that the unit dose can treat or prevent a particular disease (eg, s Μ, 佯 )) Instructions. According to the method of the present invention, a compound of the present invention which is effective for limiting or preventing a decrease in the FKBp content of RyR bound to a body (especially in cells of an individual) is administered to a body (or a cell contacting the individual). The skilled artisan can readily determine this effective dose according to conventional procedures, including titrations (titrati〇n cUrves) analysis established in vivo, and the methods and assays disclosed herein. An amount of a compound of the invention effective to limit or prevent a decrease in the amount of FKBp bound to RyR in vivo is between about 5 mg/kg/day to about 2 mg/kg/day, and/or sufficient to achieve between The amount of the compound in the plasma concentration of 3 〇〇N/ml to about 10 〇〇N/ml. In one embodiment, the compound of the invention is used in an amount between about 1 oz/kg/day to about 2 mg/kg/day. 170 200815381 Use The present invention provides a novel therapeutic method for treating patients suffering from various abnormalities associated with modulation of RyR receptors, particularly skeletal muscle abnormalities (RyRl). The present specification also provides novel therapeutic methods for treating muscle fatigue in a patient resulting from various disease states and abnormalities associated with RyR including, but not limited to, cardiac abnormalities (RyR2) and/or cognitive abnormalities (RyR3). In one embodiment of the invention, the individual has not developed symptoms of muscle fatigue (e.g., muscle fatigue caused by exercise). In another embodiment of the invention, the individual is in need of treatment for an abnormality associated with muscle fatigue, including skeletal muscle abnormalities. Various disorders of treatment or prevention by the compounds of the invention include, but are not limited to, cardiac abnormalities and diseases, skeletal muscle abnormalities and diseases, cognitive disorders and diseases, malignant hyperthermia syndrome, diabetes and sudden infant death. Cardiac abnormalities and diseases include (but are not limited to) irregular heartbeat abnormalities and diseases, irregular heartbeat abnormalities and diseases caused by exercise, sudden cardiac death, sudden cardiac death caused by exercise, congestive heart failure, and chronic obstructive pulmonary disease hypertension. Irregular heartbeat abnormalities and diseases include irregular heartbeat abnormalities and diseases caused by exercise, including (but not limited to) atrial and ventricular arrhythmia; atrial fibrillation and ventricular fibrillation; atrial frequency arrhythmia and ventricular arrhythmia; atrial frequency Pulse and ventricular frequency; catecholamine polymorphic ventricular pulsation (CPVT) and its motor-induced variants. Skeletal muscle abnormalities and diseases include, but are not limited to, skeletal muscle fatigue, exercise-induced skeletal muscle fatigue, muscular dystrophy, bladder dysfunction, and incontinence. Cognitive disorders and diseases include (but are not limited to) Alzheimer's 171 200815381 syndrome, various types of memory loss discs, 仗 仗 平 平 平 平 。 。 。 。. Those skilled in the art will be able to further identify other conditions (including but not limited to muscle and cardiac abnormalities) that may be effectively treated with the compounds of the present invention, based on the information provided herein. It is effective to limit or prevent the amount of the compound of the present invention which is reduced in the amount of FKBP12 bound to RyR1 in vivo, and is effective for preventing the amount of muscle fatigue in an individual. Those skilled in the art can readily determine the amount thereof according to conventional procedures. The procedure includes clinical trials and methods disclosed herein. Since the compounds of the present invention are capable of stably binding to FKBP of RyR and maintaining a dynamic balance of phosphorylation and dephosphorylation of protein kinase A of RyR, the compounds of the present invention are also effective for treating individuals who have experienced clinical symptoms of these different abnormalities. . For example, if an abnormal condition of the individual is detected early, the compound of the present invention can effectively limit or prevent a further decrease in the FKBP content bound to RyR in the individual. Furthermore, the individual of the present invention may be a candidate for muscle fatigue abnormalities (including #not limited to chronic diseases associated with muscle fatigue or muscle or muscle fatigue caused by stress or hard work). Thus, in yet another embodiment of the invention, the individual has been or is in motion and has developed an anomaly caused by exercise. In this case, Muss is effective in limiting or preventing the reduction of FKBP content in RyR in an individual. The amount of the compound used in the invention is an amount effective for treating an exercise-induced abnormality in an individual. Here, the servant of the present invention "effectively treats abnormalities caused by exercise" comprises a compound of the present invention in an amount of 172 200815381 which comprises an amount of the compound of the present invention which is effective for reducing or ameliorating the clinical damage or symptom of a motor-induced abnormality characterized by muscle labor. The amount of the compound of the present invention which is effective in treating an exercise caused by fatigue will depend on the specificity of each case, including the type of abnormality caused by the exercise, the body of the individual, the severity of the symptoms of the individual, and the compound of the present invention. Dosing mode. : The skilled artisan can readily determine its dosage according to conventional procedures, including clinical trials and the methods disclosed herein. In one embodiment: the prodrug compound is capable of treating an abnormality caused by exercise in an individual. The present invention further provides a method of treating muscle fatigue associated with abnormalities and symptoms caused by exercise in an individual. The method comprises administering to the individual an amount of a compound of the invention effective to treat an abnormality caused by movement in the individual. For example, an effective amount of a compound effective to treat arrhythmia caused by exercise in an individual is between about 5 g/kg/day to about 2 mg/kg/day, and/or sufficient to reach about 300 Ng/亳. Increase to a plasma concentration of about 1 〇〇〇 Nike / liter. The present invention also provides a method of preventing an abnormality caused by movement in an individual. The method comprises administering to the individual an amount of a compound of the invention effective to prevent abnormalities caused by exercise in the individual. An effective amount of a compound of the invention effective to prevent an abnormality caused by exercise in an individual is between about 5 mg/kg/day to about 20 Ng/kg/day, and/or sufficient to reach between about 3 Nike/House Increase to a plasma concentration of approximately 100 ng/ml. Further, the present invention provides a method of preventing an abnormality caused by motion in an individual. The method comprises administering to a subject an amount of a compound of the invention effective to prevent abnormalities caused by movement within the individual. The effective amount of the compound of the present invention effective to prevent the sling caused by the movement in the individual is between about 5 g/kg/day to about 2〇173 200815381 克/kg/day, and/or sufficient to reach about 300 奈The amount of plasma concentration between gram/ml and about 1000 ng/liter. The compounds of the present invention can be used alone or in combination with each other or in combination with other cardiovascular active drugs including, but not limited to, diuretics, anticoagulants, antiplatelet agents, and antiarrhythmic agents. , inotropic agents, heart rate agents (chr〇n〇tr〇pic agents), alpha and beta blockers, angiotensin inhibitors and vasodilators. Further, the above combination of the compound of the present invention and other cardiovascular agents can be administered separately or in combination. Further, one of the components of the composition may be administered before, simultaneously with or after the administration of the other drug in the pharmaceutical composition. In view of the above methods, the present invention also provides a method of using the compounds of the present invention to limit or prevent a decrease in the FKBP content bound to RyR in an abnormal candidate patient. The invention also provides methods of using the compounds of the invention to treat or prevent muscle abnormalities in an individual. Furthermore, the present invention provides a method of using the compounds of the present invention to treat or prevent muscle abnormalities caused by exercise in an individual. Intracellular Ca2+ leakage is considered to be a major regulator of muscle degeneration and atrophic muscle reorganization. Muscular atrophy is a heterogeneous hereditary disease characterized by weakness and progressive muscle atrophy. All forms of muscular dystrophy involved in the dystrophin-associated protein complex, also known as dystrophinopathies, are the most common form of Duchenne muscular dystrophy (DMD). One of the genetic diseases (sex 174 200815381 singularity before the age of a reduced or meat atrophy and shell cell growth, suffering from high-systolic heart disease, atrophy Duchenne muscle mainly caused by the increase of Ca2 + - According to the mouse and calcium meat atrophy calcium-dependent, one out of 3,500 children), and most patients usually die of respiratory and/or heart failure at 3 weeks. Bayesian muscular dystrophy (BMD) is a mild form of disease that is associated with the performance of truncated myosin in dystr〇phin, but is completely absent or minimal in patients with Duchenne. The content of myosin. Duchenne muscular atrophy (DMD/BMD) is caused by a mutation in the gene encoding 427-kDa fine protein-myotrophin. However, with age, patients with Bayesian muscular dystrophy have more common visceral symptoms than those with Duchenne muscular dystrophy and are not associated with musculoskeletal symptoms. Due to the incidence of confinement, it is not possible to remove Duchenne muscles by genetic screening. Therefore, an effective treatment is needed. It is known that Duchenne and Bayesian muscles are associated with abnormal metabolism of intracellular calcium. Since it is generally believed that the change of Ca2+ concentration in muscle fiber cells of patients with atrophy is a disease mechanism, it is urgent to develop a treatment for preventing Ca2+ abnormality in skeletal muscle degeneration. The main genetic defect of Duchenne and Bayesian muscular dystrophy is the lack of muscle performance. However, it is still unknown that the intracellular Ca2+ concentration ([Ca2·^) will directly cause damage to harmful myocytes (muscle fibers) and simultaneously activate lysin 6# ( Ca2+-dependent proteases). Activation of proteolytic dysfunction due to elevated calpain activity in wdx necrotic muscle fibers will lead to limb-type disease. Therefore, prevention of activation of sputum protease by inhibiting intracellular Ca2+ concentration can be used as a preventive for Duchenne muscular atrophy. Patients with muscle wasting 175 200815381 A method of contraction. Significant differences in abnormal and atrophic muscles have been observed in the myotubes and the hearts of the animal 4 myosin. The increase in intracellular Ca2+ concentration is prevented by administration of a compound containing the present invention. The compounds of the invention may be prepared in various forms, as well as the salts of the compounds, solvates, complexes, prodrugs or prodrugs, including all of the different formulas of the compound. The invention further provides for the inclusion of a radiolabeled present invention. The compounds of the invention are labeled using a variety of different radiolabels known in the art. For example, the radioisotope of the present invention. The isotope is any radiometric measurable, including (not limited to) 35s, 125i, 3h or 14c. A method can be used to detect the radioactivity emitted by the isotope. For example, 'technique (especially referred to as scintigraphy) detects T-rays of isotopes<by way of non-limiting example, the standard compound is prepared in the following manner. The base of the compound of the present invention is utilized as B B r 3 . Next, in the presence of a base (eg, NaH), a methylating agent (eg, a 3H-sulfuric acid phenolic compound is remethylated to provide a 3H-labeled pseudonym using forced swimming as a booster mouse bone The muscle has an effective program to study the state of the skeletal muscle RyRl channel complex. Unexpectedly, swimming 90 minutes twice a day for up to 3 weeks Mo (including lack of meat between [Ca2, pharmaceutical compositions such as salts, water, and The method for grouping the compounds of the present invention, wherein the radioactive label is a radiation of the same substance, which is commonly used in the art, 7 angiography technology is used to remove the methyl ketone methyl ether from the radioactive benzene ring of the invention) After the component of oxygen motility and phosphorylation, C57B16 176 200815381 wild-type mice showed significantly increased RyR2 acidification (by protein kinase A) whereas Ca2, cadherin kinase II (CaMKII) phosphorylation did not change, Point out the clarity of the pressure pathway, in which the RyR 1 protein is stable, but the RyR1 channel lacks the stable subunit calstabinl (FKBP12). RyRl hyperphosphorylation and lack of calst have been shown Ablin is consistent with the leaky RyRl channel that causes intracellular Ca2+ leakage. After swimming for 90 minutes twice for up to 3 weeks, the RyRl channel is hyperphosphorylated by protein kinase A and lacks the stable calstabinl subunit. The RyR 1 macromolecular channel complex showed an increase in protein kinase A phosphorylation of Ser-2 844 (corresponding to human RyRl-Ser-2843), whereas CaMKII phosphorylation of Ser-2849 (corresponding to human RyRl-Ser-2848) did not change. With the increase in RyRl-Ser-2844 protein kinase A hyperphosphorylation, the channel complex begins to lack calstabinl. Phosphorylation and normalization of the four subunits of the tetrameric channel complex can show protein kinase A filling. A significant increase in acidification and a lack of stability of the calstabinl subunit. Example 1: Effect of S36 A common name for the RyCal compound named S36, S107 was synthesized as described in PCT/US2006/32405, as described in the co-pending application USSN 1 1/212,309. Figure 6 shows some of the molecular mechanisms that cause muscle fatigue and the effects of S36 on muscle fatigue. : The same litterate eight-week-old wild-type C57BL/6J mice were randomly received 36 or Carrier treatment of either. 177 200815381 Two days before the start, a horizontal incision in the neck of the neck will be filled with 200 ul of phosphate buffered saline (PBS) or 2 〇〇W of S36 ( Diluted in PBS at a concentration of 10 pg/ul) osmotic irrigation / 隹庄帮浦 (Alzet Model 2004, Durect, Cupertino, CA; total 穑1 槚20〇μΐ, delivery rate 0.25 μΐ / hr) subcutaneous Implanted into the back surface of the mouse. The mice were allowed to recover for three days before starting exercise. Standard food and moisture are supplied arbitrarily.运动动: Starting on the first day, Xiaoyan spleen, silkworm red scorpion soil J will exercise for up to 3 weeks; swim 5 days a week and run 1 hour a week on the treadmill.

View-style: - The daily swimming program consists of two swimming sessions during a one-hour break every day. The initial training method for up to 5 days is increased by 1 minute from 4 minutes to 80 minutes, and each swim course lasts 90 minutes. An opaque acrylic water tank 30 cm long and 30 cm wide is filled with tap water to a depth of at least 20 cm. The water is circulated by a separate water storage tank with heating elements, a thermostat and a pump to heat the water to 32_34 〇c. The Tyg0n pipe, which has a small needle-shaped hole at the bottom of the tank, emits compressed gas to agitate the horizontal plane. Paired with the treatment population (4 pairs of appropriate mice with pair-wise ~ 丨 11 可 can swim in the sink at any one time point. The same litter but not exercised mice were retained as a negative control group. Each individually recognized mouse swimming event utilizes a video tracking system (San Diego Instruments, San Diego, CA) that includes a Sony CCD video recorder, a DVD/Hard Drive, an image capture card, and a good condition. The SMART 2.0 software customized for the social behavior package of 8 mice can be tracked at the same time. The mice were anesthetized with oxygen 178 200815381 1.5% isoflurane in the gas, and the small 0.75 cm micro-can be used. The Velcro coin is sewn to the scalp of each mouse with a 5.0 nylon suture. The 1 cm colored plastic dot adhered to the fishing surface side of the Ve 1 cr 〇 (h ο 〇kside) can be firmly attached to Multiple body tracking on mice and under appropriate light conditions. Analysis of mouse results tracked by X-axis, Y-axis and time and averaged 2, 5 and 10 minute intervals and swimming distance .

The Instruments (Columbus, 0H) treadmill (model: Exer-6M Treadmill with Treadmill Shock Detection Unit) allowed the mice to run. The mice were placed on their respective runways at the lowest rate (7 m/y) and the sh〇cking apparatus was turned off and allowed to acclimate to the environment for 6 minutes. The front half of the treadmill is covered with aluminum foil to block light. The table lamp illuminates the scared area at the back end of the treadmill. k should be P after the segment, turn on the current and record the number of frights sent during the next two three-minute intervals (training = period). Then reset the startle counter, lift the speed to 10 m/min, and observe the scared area and record loss at three-minute intervals: the fright of each mouse until the end of the experiment. The distance of ..., step = from the initial 〗 〇 / / min rise to 36 meters, minutes ... minutes speed ==: over 2 meters, points. Apply to all mice of a specific scorpion - high. Going, the difficulty of the program on the experimental schedule has been picked up, avoiding the scare area and giving up or when the mouse has anointed you as a mission failure. In almost all cases, the two are almost the same. 179 200815381 Separation of plastic seats: After the 21st and 2nd days of exercise, the mice were given the last swim with an unbelievable curriculum. After 9 minutes of swimming, each mouse was sacrificed immediately by inhaling carbon dioxide and cervical dislocation (cervical disl〇cati〇n). The blood is removed by centrifugation in the heart and centrifuged, and the plasma is eluted and frozen in liquid nitrogen. Two toe longissimus muscles were exposed and wetted with Tyrode's solution, and a 4.0 thread was tied to the proximal and distal tendons, and the muscles were freely cut. Perfuse muscles with 2.0 mM CaCl2 and 100% 〇2 bubbling and heating to 35 °C, and suspending the muscles in isoletric force transducers (F-30) , Harvard Apparatus, Cambridge, MA). The force-frequency relationship was measured after 1 cN resting tension was balanced for 10 minutes and a short-term gaining scheme with a 60 second delay between 800 ms stimulation at 40-1 50 Hz. Fatigue is generated with a tonicity regimen of 50 Hz every two seconds (600 ms each) for 120 seconds. DMC ν4·1·6 (Aurora Scientific, Canada) was used to stimulate and record muscle response, and DMA v3.2 (Aurora Scientific, Canada) was used to analyze the data. After the stimulation, the muscle length was measured under resting tension and the dry weight was recorded. A long toe long muscle was frozen in isopentane (·8 (TC ) for histological analysis and the other was frozen in liquid N 2 for biochemical analysis. In addition, two soleus muscles were cut and the same One was frozen in isopentane for histological analysis and the other was frozen in liquid N2 for biochemical 180 200815381 analysis. The outer muscles (vastus lateralis), heart and diaphragm were also cut from each animal and frozen. Used in liquid N2 for biochemical analysis. Biochemical analysis: inhibition with 0.5 ml buffer (50 mM Tris HC1 buffer, pH 7.4, 0.9% NaCl, 5.0 mM NaF, 1.0 mM Na3V04, 0.5% Triton X-100 + protease) The RyR antibody in the agent was immunoprecipitated from the skeletal muscle homogenate at 4 hours (the culture was followed by Protein A sepharose beads (Amersham Pharmacia) at 4 Incubate for 1 hour at ° C, after which the beads were washed three times with buffer. To detect RyR2, the protein was separated on a 6% sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE), and Calstatab2, protein on 15% SDS-PAGE Divided in size and transferred to nitrocellulose membrane (SemiDry transfer blot, Bio-Rad) overnight. Cultured in 5% skim milk to avoid non-specific antibody binding and in Tris buffered saline solution with 0.1% Tween-20 ( Tris-buffered saline) After washing, the membrane was incubated for 1 _2 hours at room temperature with the following primary antibody: calstabin antibody (1:1,000); RyR antibody (5029; 1:5,000); or RyR2-pSer2809 phospho-based antibody ( 1:5,000), which detects a protein kinase A-mediated mouse RyR 1 -pSer2844 and RyR2-pSer2 808 〇 after three washes with wasabi peroxidase-labeled rabbit IgG antibody (1:5,000, Transduction Laboratories) , Lexington, KY) cultured membranes and developed with an enhanced chemiluminescent extraction system (Amersham Pharmacia). Band density was quantified using QUANTITY ONE software (Bio-Rad) 181 200815381 Example 2 · Figures 7-20 of S107 show the molecular mechanisms and effects of muscle fatigue. Drug delivery: C57BL/6J mice of the same litter size of eight weeks of age were randomized to receive either S107 or vehicle (H2). Three days before the start of each test, 2 ul of phosphate buffered saline (PBS) or 200 μM of S107 (diluted in H 2 O at a concentration of 10 μβ/ι1) was placed through a horizontal incision on the neck. Permeation perfusion pump (Alzet Model 2004,

Durect, Cupertino, CA; total volume 200 μl, delivery rate 〇 25 μΐ/hr) was implanted subcutaneously into the dorsal surface of the mouse. The mice were allowed to recover for three days before the start of the exercise. Standard food and moisture were arbitrarily supplied throughout the experiment. Long-term exercise mode = The daily swimming program consists of two swimming sessions during a four-hour break every day. Up to 5 days of initial training The swimming course is increased by 1 minute from 40 minutes to 8 minutes, and each swim course lasts 90 minutes. An opaque acrylic water tank 30 cm long and 30 cm wide is filled with tap water to a depth of at least 20 cm. Use a separate water storage tank with heating element, thermostat and pump to circulate and heat the water to 3 2 - 3 4 °C. Eight mice paired with the genotype and/or treatment population can swim in the sink at any one time point. Mice that were born in the same place but did not exercise were retained as a negative control group. In order to confirm that a consistent exercise environment was achieved, a pilot experiment using a video tracking system (San Diego Instruments, San Diego, CA) to track the movement of each individual 182 200815381 was performed. Use the SMART 2.0 software with Social Behavior package (San Diego Instruments) to analyze the individual records of each 90-minute swim track to get the distance traveled and the average speed from start to finish. There was no significant difference in the degree of exercise. Tread_Car Performance: The mice were run using a Columbus Instruments (Columbus, OH) treadmill (Model: Exer-6M Treadmill with Treadmill Shock Detection Unit) with six runways. Turn off the scare device and place the mice on their respective runways and let them adjust to the environment for 1 〇 minutes. The front half of the treadmill is covered with aluminum foil to block light, and the table lamp illuminates the scared area at the rear of the treadmill. After the adaptation phase, the treadmill was set at 1 cm/min and the mice were trained to run for 6 minutes with mild stimulation. The current is then turned on and the number of intimidation delivered during the next two three minute intervals (training period) is recorded. The frightening counter was then reset and the scare area and the shock of the record delivered to each mouse were observed at three minute intervals until the end of the experiment. At a fixed distance, the treadmill speed increased from the initial 10 m/min to 24 m/min. The speed increase will not exceed 2 m/min every 6 minutes. When a mouse is unable to continue running (even if it is slightly irritated), it is defined as a mission failure. Immediately after the complete muscle specimen L forced exercise course, each mouse was sacrificed by inhalation _ carbon monoxide and neck dislocation. The blood was removed by centrifugation in the heart and centrifuged to elute the plasma and cool the beads in liquid nitrogen. Attach the silk thread of 4·〇 to the tendon near the distal and distal ends of the long toe and the soleus muscle, and cut the muscle freely and place it in a modified forest with a bubble of 100 〇/〇〇2 183 200815381 Ringer's solution (140 mM NaCl, 5 mM KC1, 2.0 mM CaCl2, 2 mM MgCl2, 10 mM HEPES, 10 mM glucose, pH 7.4). The muscles were suspended vertically in a 50 mL Radnoti jacketed glass chamber, and one tendon was attached to the isotonic contraction force transducer (F-30, Harvard Apparatus, Cambridge, ΜΑ) with 4.0 threads, and the other The tendon is attached to the fixed arm of the built-in platinum irritating electrode plate with a suture. The force-frequency relationship was measured after the muscles were filled with 3 5 QC Ringer's solution and equilibrated with 1 cN static tension for 10 minutes with a short-term potentiation protocol, between 800 ms and 40 ms at 150-150 Hz. Has a 60 second delay. The muscle response was stimulated and recorded using DMC ν4·1·6 (Aurora Scientific, Canada), and the data was analyzed using D M A v 3.2 (A u r 〇 r a Scientific, Canada). After stimulation, muscle length was measured under static tension and muscle dry weight was recorded. Conjugated focal microscopy: Fibro digitorum brevis (FDB) fibers were isolated by standard method enzymes (Reiken, Lacampagne et al. 2003). Briefly, the muscle is cut from the paw and placed in a modified Ringer's solution, and all fascia is removed. Type 1 collagenase (2 mg/ml, Sigma) was freshly placed in Ringer's solution, and the muscle was decomposed for two hours in a 37 ° C incubator (shake at 1.25 rpm). Place the muscles in fresh Ringer's solution and carefully grind them. A single fiber was collected and attached to a cover glass of laminin (Sigma L-2020). The cells were loaded with 2 μΜ fluo4-AMg (Invitrogen) for 20 minutes, placed on a Zeiss Live 5 microscope stage and filled with Ringer's solution for 15 minutes. The fiber enthalpy was adjusted at i Hz frequency before the presentation of the basic properties of the fiber 184 200815381. The linear scan image is continuously taken at a scan rate of 1 ms at a linear order of 300 s s z stimuli (with a 2 Hz training speed) at a frequency of 1 0 0 Η z. The images were analyzed in ImageJ and the F/FO ratio of each fiber was calculated. Human Exercise Program: Three weeks prior to the pilot course, the Individual to the Human Performance Laboratory (Appalachian State Univeristy) described the basic measurements of cardiorespiratory fitness and body composition. On three consecutive trial day, the individual consumed standard breakfast (7-8:00 in the morning) and lunch (before 12:30 pm), followed by an overview of the ASU Human Performance Lab at 2:00 pm. Individuals move on a sports bike with a 70% V02max from 3:00 pm to 6:00 pm. The test course day is Monday, Tuesday and Wednesday afternoon. Oxygen consumption and other metabolic parameters were measured every 30 minutes using an indirect calorimeter with a mouthpiece and nose clip, and blood lactate and glucose levels were measured every 60 minutes (via fingertips) to confirm individual compliance The amount of exercise. Individual movements drink 5-1·0 liters of water per hour while avoiding all forms of inhaled energy (eg, alcohol, beverages). Individuals in the resting control group were sitting in the laboratory during the exercise test. Each of the three trial sessions collected ash, urine, and saliva samples 15-30 minutes before exercise/quiet sitting and then 5-10 minutes after exercise. Muscle section samples were taken on days 1 and 3 using the needle bi〇pSy procedure 15-30 minutes before exercise/quiet sitting and then 5-10 minutes after exercise. Take four samples at intervals of about 2 inches (two out of each strand). The sections were snap frozen in liquid nitrogen and staggered at -80 °C. 185 200815381 Single channel recording data acquisition: Preparation of sarcoplasmic reticulum vesicles from sedentary mice and long-term exercise mice (treated with either vehicle or S107) as previously described (Reiken, Lacampagne et al. 2003) ). The RyRl channel is reconstituted by spontaneous fusion of microsomes into planar lipid bilayers. The ratio of phosphatidyl ethanol amine to phosphatidylserine is 3 Mixture of :1 (Avanti Polar Lipids). The planar lipid bilayer membrane was formed in a 200 μιη hole in a polysulfonate cup (Warner Instruments, Inc.), which separated the two soaks (1 mM EGTA, 250/125 mM HEPES/Tris, 50 mM KC1). , 0.5 mM CaCl 2 , pH 7·35 as the ipsilateral (ch) solution and 53 mM Ba(0H) 2 , 50 mM KC1, 250 mM HEPES, pH 7.35 as the reverse solution). After the incorporation, the RyRl channel activity was continuously recorded for at least 10 minutes. The free Ca2+ concentration in the cb compartment was calculated using the WinMaxC program (version 2.50) (Bers, Patton et al. 1994). A single channel current at 0 mV was recorded in a gap-free mode using an Axopatch 200A patch-clamp amplifier (Axon Instruments, USA), filtered at 1 kHz and digitized at 10 kHz. Data acquisition was performed using the Digidata 1322A and Axoscope 9 software (Axon Instruments, USA). The records were analyzed using Clampfit 10·1 (Molecular Devices, USA) and Origin software (version 6.0, Microcal Software, Inc., USA). Analysis of Argonine Receptor Etiquette: Isotonically dissolved 10 mg muscle samples. By 250 gg homogenate with 0.5 ml modified 186 200815381 RIPA buffer (50 mM Tris-HCl (pH 7.4), 0.9% NaCl, 5.0 mM NaF, 1.0 mM Na3V 〇 4, 1.0% Triton-Xl The RyR antibody (2 μΐ 5 029 Ab) in sputum and protease inhibitor was incubated at 4 ° C for 1 hour to immunoprecipitate the RyR receptor RyR 1 . The samples were then incubated with Protein A Sepharose microbeads (Ammersham Pharmacia) for 1 hour at 4 ° C, after which the beads were washed three times with buffer. The protein was separated on a 4-20% concentration gradient of sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) and transferred to the stone pathological fiber membrane overnight (SemiDry transfer blot, Bio-Rad). ). After incubation with a blocking solution (LICOR Biosciences, Lincoln NE) to avoid non-specific antibody binding and washing in Tris buffered saline solution with 0.1% Tween-20, the membrane was incubated for 1-2 hours at room temperature with the following primary antibody: Calstabin antibody (1:2500 in a blocking solution); RyR antibody (5029; 1:5,000); or RyR2-pSer2809 phospho-based antibody (1:5,000), which detects protein kinase A phosphorylated mouse RyRl-pSer2844 With RyR2-pSer2808; and PDE4D3 antibody (1:1 000). After three washes, they were incubated with infrared-labeled secondary antibody (1:10,000 dilution, LICOR Biosystems). Band density was quantified using an Odyssey Infrared Imaging System (LICOR Biosciences). Dance-activated chymase and creatine kinase assays: Tissue #5-activated protease activity was measured using a calcium activating protease activity test kit (C a 1 b i 〇 c h e m, Sa n D i e g 〇, C A). This test was based on the decomposition of the fluorescent peptide matrix Suc-LLVY-AMC (Calbiochem). The muscle homogenate was diluted to a final concentration of 600 pg/ml and the activity of calcium activating protease in the plasma of 187 200815381 was measured according to the manufacturer's instructions. Plasma creatine kinase (CK) activity was tested using a kit of reagents (Pointe Scientific, Inc., Canton, MI). Plasma samples (each 5 μl copy) were added to 200 μl of the creatine kinase reagent and the change in absorbance at 340 nm for 4 minutes was recorded using a plate reader. The change in average absorbance per minute was used to determine the degree of CPK according to the manufacturer's instructions. The analysis presented the data as a mean standard deviation (SEM). An independent experiment with a significant degree of 0.05 was used to test the difference between cair/_ and wild type, PDE4D small and wild type, and exercise + carrier versus exercise + si〇7 except for the following. The distribution of treadmill failure data is found to be asymmetrical in many instances. Therefore, the Wilcson x 〇 rank rank test is used to compare all such data. Also intensive to cause RyRl protein stimulating A hyperphosphorylation | You - Tong 4 agricultural complex U calstabinl Austrian PDE4D3: The movement patterns in mice have been divided into two types: υ autonomous movement, including the running wheel Circle; and 2) involuntary movements, including swimming or forced treadmills to exhaustion. In order to achieve high-intensity exercise, the twice-daily swimming program is designed to achieve consistent exercise in mice for days to weeks. This mouse exercise regimen is not designed with either centrifugation or isotonic characteristics, but rather a physiologically mixed type that is centrifuged into isotonic motion. After forced exercise, RyRi was immunoprecipitated from the entire muscle of the hind limb muscles, and the RyR1 channel complex was separated by size and directed against the channel on sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE). The complex components were subjected to immunological dot analysis. High-intensity 188 in 200815381 degrees of exercise caused RyRl to gradually phosphorylate at the protein kinase A site Ser2844 (RyRl-pS2844), and this phenomenon was performed twice a day (9 minutes course, eg Figure 8A) for 14 days. When it reaches saturation. In addition, the RyRi macromolecular complex of the long toe longus muscle (EDL) was recombined, including the lack of calstabinl and PDE4D3 (Figures 8A and 8B) for nearly 14 days. The same biochemical alterations were observed in all other hind limb skeletal muscles isolated from the same mouse, including the soleus muscle, tibialis anterior and gastrocnemius. RyRi protein kinase A hyperphosphorylation and lack of caistabinl and PDE4D3 depend on exercise intensity, while only relatively high intensity exercise can cause significant channel changes (8C and 8D). Furthermore, recombination of the RyRl channel complex continued after exercise and only partially recovered after three days of rest after long periods of exercise (Fig. 16A). Protein kinase A hyperphosphorylation of RyRl is not altered by the amount of protein kinase A and PP1 bound to the RyRl complex, as these levels are not affected by the state of motion (Figures 16A and 16B). During exercise, there was no change in the calstabinl content in the whole muscle pulp measured by immunoblot analysis (Fig. 16C). Therefore, high-intensity exercise for several weeks per day causes recombination of the RyRl channel complex's main: channel lacking phosphodiesterase PDE4D3; protein kinase A hyperphosphorylation; and RyRl channel complex lacking stability subunit ealstabinl ° human RvRl channel sensitivity in exercise sputum in order to verify whether the recombination of RyRl channel macromolecular complexes found in exercised mice is significant for human physiology, in high-intensity exercise programs (57% of 189 200815381)

V〇2m ax bicycles for 3 hours/day) Human muscles were taken from trained athletes before and after the first and third days of exercise (Nieman, Henson et al. 2006). Human RyRl macromolecular complex was immunoprecipitated from muscle homogenate and separated by SDS-PAGE and immunoblot analysis to evaluate RyRl protein kinase A. The degree of phosphorylation and the content of calstabinl and PDE4D3 in RyRl complex. High-intensity exercise caused excessive proteination of RyRl's protein kinase A and lacked cal stab ini (compared to the non-exercised control group) (Fig. 9). Before the third day of exercise, the RyRl protein kinase A phosphorylation system in the trained bicycle knight was at or near the stationary stage, and the RyRl complex was not found to be significantly lacking calstabinl, however, at the beginning of the third day of high-intensity exercise The RyRl complex stably lacks PDE4D3 (Fig. 9B). Therefore, the same recombination of the RyRl channel complex found in long-lived mice can occur in highly trained athletes who receive intense exercise. Muscle-specific calstabinl^ mice have high-intensity motor deficits: muscle-specific loss of calstabinl has previously been shown to be associated with changes in the force-frequency relationship of isolated muscle samples and reduction in C av 1 · 1 current (Tang, Ingalls et Al· 2004). In order to determine calstabinl binding to

Whether RyRl has an effect on athletic performance, the inventors assessed muscle specificity and lacked calstabinl mice to run to exhaustion. The call·plant mice had significant defects in high-intensity exercise ability relative to the wild-type control group of the same litter (Fig. A). The motor deficits in males and females are similar (Fig. 10B), and although there is a slight decrease in body 190 200815381 in call·/· mice (1 oc map), there is no association between failure time and weight loss. (10D alone). Motor deficits are most pronounced in high-intensity exercise (tachometer speed equal to or at 24 m/min) or centrifugal motion (for example, a downhill treadmill run of 14.). 0/2 call-/- mice were able to complete a 30-minute downhill treadmill program, but 2/2 wild-type mice of the same litter could complete the program. 24 hours after the downhill movement (as described herein), plasma creatine kinase (CPK) increased in the ca/plant mice (relative to the wild-type control group of the same litter) as muscle damage increased (Fig. 10E). After exercise for several days after exercise, the ability of wild-type mice to move close to ea// - the ability of eight mice to move may be due to the gradual lack of 4 RyRl complexes... heart-! (long-term exercise in wild-type mice) (in Figure 8), the RyRl-passing C complex caused by exercise lacks caistabini, which is similar to the phenomenon found before the movement of mice (p. 10F). Therefore, the lack of muscle specificity calstabinl 窜 ^ & shows that fatigue increases with the lack of calstabinl in the RyRl complex, whereas the upper 丄 L _ @ plays a role in muscle fatigue. PDE4D + mouse two-one-dynamic defects: systemic lack of PDE4D causes age-dependent progressive cardiomyopathy (Lehnart, Wehrens Μ & 2005). However, before 3 months of a/), PDE4D-/. mice did not show a deficiency in the heart that could be measured by ~scene supercavitation or catheter; cardiac catheterization. The exercise capacity of two-month-old PDE4D·/- mice can be compared to wild-type mice that are homogenous. A significant decrease in exercise capacity (Fig. 3) can be found and is not associated with body weight (graphs iib, c, d). 191 200815381 Obviously, the high CPK content of resting PDE4D·/- mice g (5) (relative to the wild-type control group of the same litter) is consistent with muscle injury, j field damage, and single centrifugation of PDE4D+ mice (by 30 minutes, Xiachengcheng consists of running on a treadmill.) The CPK content is significantly improved 24 hours after the insertion (Division p p E). P D E 4 D / mice after only one day of light exercise showed that they lacked the surname, RyRl combined with PDE4D3, basic

A slight increase in phosphorylation of RyRl protein kinase A, as well as a marked increase in deficiency (Fig. UF). These data show that R.R. plays an important role in the RyR1 complex by regulating the degree of phosphorylation of protein kinase A by RyRl, whereas the lack of pDE4D3 during exercise-time channel complex phosphorylation promotes protein kinase A, which in turn contributes to muscles during exercise. Injury and skeletal muscle fatigue. Complexology combined with calstabinl in RVR1 can improve the county time table ^2 has proved that the lack of calstabinl is related to motor defects and long-term or two-intensity exercise can cause RyRl lack of calstabinl, also confirmed pharmacologically combined calstabinl in RyRl pair The effect of long-term or high-intensity exercise performance. The 1,4-benzothiazepine derivative (RyCal compound) was screened to find high target activity, high specificity (no activity against other known ion channels) and in vivo efficacy in improving exercise performance. Compound S107 was found to not affect L-type calcium channel current or hERG potassium current at 500 nM. Age- and sex-matched wild-type mice will be randomized to receive a osmotic pump containing either si 〇7 or any of the carriers. Before the start of the 21-day forced swimming mode, a dose of 2.5 ug/hour of S107 or vehicle was used to inject four 192 200815381 days. Treadmill running with the exhaustion of the mouse at night and in the exhaustion can hinder the assessment of exercise capacity once. Mice will not exercise on the same day as the treadmill assessment. Figure 12A shows that calstabinl recombination caused by S107 does not have a serious effect on wild-type exercise performance, but wild-type mice treated with S 1 〇7 are more resistant to treadmill exercise ability over time (occurring in Vehicle-treated mice) (Day 21st S 1 0 7 relative vehicle, Wilkerson assay p < 〇〇 ^' These studies are due to the training effect of repetitive motion (performance due to increased muscle tissue) It is complicated and complicated. Figure 2B shows the time of the treadmill failure on the 21st day. In the field stimulation, the toe in the tissue bath is in the tissue bath. The long toe long muscles of the isotonic S1 07 treated mice in the long muscle showed an increase in power generation at a stimulation frequency higher than 8 Hz, which corresponds to the left shift of the force-frequency relationship map (12C) Figure). Drug treatment does not cause weight (Fig. 12D) or changes in muscle mass. In similar long-term exercise tests in which mice lack calstabinl, S107 does not improve treadmill performance (Fig. 12E). Long-term exercise regimen a lot of RyRl Protein kinase A phosphorylation and the lack of calstabinl in the RyRi of the immunization hall. S107 treatment almost completely reverses the phenomenon that calytabin is absent in RyRl (Fig. 12F). Putting these data together can be shown to improve calstabinl to channel-bound drugs to prevent Because protein kinase A hyperphosphorylates the sarcoplasmic reticulum Ca2+ leakage caused by the RyRl channel, it can prevent muscle damage, muscle fatigue, improve muscle function and improve exercise performance in a fatigue program. 193 200815381 Recombination with fatigue in the short toe Destruction: After long-term exercise regimen, the fibroblast short muscle (FDB) fibers were separated from the mice by enzymes and loaded with the calcium ion indicator flu〇_4. The electric field stimulation at 1 Hz was in the fatigue program (by every two In the process of 300 ms long repetitive type, 120 Hz tonic stimulation for up to 400 seconds), each muscle fiber was presented on a zeiss Live5 conjugated focus microscope. Vehicle-treated mice were shown (Grade 3 A) Representative F/F0 traces of flexor short muscle fibers isolated from S107 treated mice (Fig. 13B) during the fatigue protocol. The short flexor fibers of si 07 treated mice showed rigidity #5 Ion transient current peak drop delay (Fig. 3 C). It is known that muscle fibers with slower Ca2+ release and resorption kinetics are less prone to fatigue. Evaluation of Ca2+ release during single contraction at 1 Hz frequency Resorption kinetics. The distribution of 50% resorption time (τ) was not significantly different between vehicle and S107 treatment (Fig. 17), indicating that there was no change in the calcium resorption kinetics of flexor short fibers. These data indicate that treatment with S107 improves Ca2+ management in muscle fibers during fatigue regimens. The leakage of the RvR1 channel caused by prolonged exercise can be reversed by the binding of calstabinl: a key debate is whether the biochemical changes found in the RyR 1 macromolecular complex during exercise cause a change in RyR 1 channel activity. To directly investigate this problem, sarcoplasmic reticulum microsomes were prepared from hind limb muscles of the following mice: sedentary mice, mice that were exercised for a long time and treated with vehicle, and small animals that were exercised for a long time and treated with S 107. mouse. The blister was fused into a planar lipid bilayer membrane using standard techniques and the single channel activity of the RyRl channel was continuously measured for up to 10 minutes at 90 nM [Ca2 + ]e/i (Fig. 14A). Consistent with previously published data (Meissner, 1994, Reiken, 2003), sedentary 194 200815381 mice's RyRl activity is very low at the stationary helper calcium ion concentration, resulting in only a few open in long-term time (in, In the experiment, a record of up to 20 points is required to calculate the opening period of the channel). In contrast, the RyR1 broadcast channel of mice that were exercised for a long period of time and treated with vehicle showed an increase in activity and a significantly higher rate of opening (sports + vehicle group (n = , 19) relative to sedentary group (n = 9) , p < 0.005), this is due to the opening frequency of the ancient, the two hands of the hand (Figure 14). S107 was applied to cause a significant decrease in RyRl opening rate (human sway + S107 group (η = 12) relative motion + carrier (n = 9), ρ < 0.00 5) to 盥 # J master 1 and sedentary mouse channel Comparison process

degree. Long-term exercise or S107-free has no effect on the length of the channel, which means that the change found in the open rate is caused by the change in the number of open events (Fig. 14B). These data show that the RyRi channel of the sport animal exhibits a "leakage" channel behavior (high opening rate) and the channel of the animal treated with sl7 is not "leakage type". C_alstabin 1_再I.·Coincided to reduce the activity of activating proteases in muscle tissue and reduce the damage of muscle tissue: a possible mechanism of ionic damage in the release of RyRl channel is a Ca2+-dependent neutral protease calcium activation Activation of a member of the protease family, whereas Ca2+-dependent neutral proteases are known to be the cause of muscle damage in many physiological conditions (Belcastro 1 993; Berchtold, Brinkmeier et al. 2000). The calcium-activated protease activity in the muscle homogenate was evaluated by decomposition of the synthetic calcium-activated protease matrix Suc-LLVY-AMC (fluorescence upon calcitonin cleavage). The prolonged toe long muscles of long-term exercise showed higher calcium activating protease activity than the sedentary control group. Calcium-activated protease activity was significantly reduced in S 1 07 treated and long-running mice (Fig. 15A). A further step by 195 200815381 measures the degree of plasma CPK activity to provide evidence of protection against muscle damage, with elevated levels of plasma CPK activity in prolonged exercise mice, but reduced to near sedentary controls in S107 treated mice. The degree (Fig. 1 5 B). Muscle histology showed evidence of muscle hypertrophy in long-moving mice with some inflammation and impaired fiber-distributed distribution, while there was no evidence of massive cell necrosis in any of the treatment groups. ϋ 3 : Muscle atrophy and the effect of S107

The RyRl calcium ion release channel becomes hyperphosphorylated (caused by protein kinase A) and lacks the stable protein calstab ini during exercise. The compounds of the invention increase the binding affinity of cal stab ini to RyRl, which is hyperphosphorylated by protein kinase a. These compounds (referred to as "calcium channel stabilizers" or "RyCal") are 1,4 -benzothiazepines and their derivatives. Treatment with these compounds improves the performance of mice running on a treadmill. Calcium ion leakage through the RyRl channel, which is over-acidified by protein kinase A, causes muscle damage caused by calcium-dependent protease activation, while RyCal prevents word leakage and inhibits muscle damage during long-term exercise. RyCa] can be applied to improve muscle fatigue in chronic diseases including heart failure, AIDS, cancer, and kidney failure and can be used to treat muscular dystrophy. Duchenne muscular atrophy (DMD) is a muscle disease of X-Hnked, characterized by mutations in the myosin gene. The increase in calcium-activated calcium-activated protease protein hydrolysis in the sarcolemma membrane is considered to be the main mechanism of Duchenne muscular dystrophy physiology 196 200815381. In the 23rd exon of the myosin gene, a stop coding for ^^mdx mice was provided, providing an effective study of the effects of different treatment strategies on the treatment of this disease. A RyCal compound, S107', reduces mi/jx: calcine-activated calpain activity (Figs. 7, 18, 19, 20). This indicates that RyCai is used to treat muscle-related diseases (including, but not limited to, muscular dystrophy). Animal model: Ux mice (21 days old) were treated with an implantable osmotic pump with Sl7 (〇 125 mg/kg/hr) or vehicle for up to 2 days. After treatment, the mice were subjected to a downhill (14°) treadmill running at 18 m/min for 30 minutes. Immediately after the exercise, the animals are sacrificed and the skeletal muscles are obtained. Preparation of the toe long muscle homogenate: The toe long muscle homopolymer was prepared in 0.5 ml of homogenization buffer (20 mM NaF, 10 mM Tris-maleate, ρΗ7·2 + protease inhibitor). The heart sarcoplasmic reticulum (CSR) fraction was obtained by centrifugation at 50,000 xg for 30 minutes. The slurry was homogenized at 4000 xg for 20 minutes and the supernatant was centrifuged at 10,000 x g for 20 minutes. For the protein concentration test, an equal amount of the sample (Aliquot) was stored and stored at -80 °C. Calcium- and i-protease bioassay: Calcium-activated protease activity assay kit (Calbiochem) was used to measure the calcium-activated protease activity of the toe-long muscles (3 μ μ§). This assay utilizes the synthetic calcium activating protease matrix suc-LLVY-AMC. Calcium-activated protease cleaves amyl and is measured by fluorescence. The assay was performed with both activation and inhibition buffers to determine the definitive #5 activating protein intoxication activity in the sample. 197 200815381 Serum creatine kinase: serum (10 μΐ) protein was separated by 4-20% PAGE. After transferring the protein to the nitrocellulose membrane, the immunoblots were developed using a creatine kinase antibody (Research Diagnostics, 1:1000 dilution). Bands were quantified using densitometry. Immunoprecipitation of the Arino base receptor: 250 samples of the long toe muscle of the toe and the RyR antibody (2 μΐ 5 029 Ab) were co-cultured at 4 ° C for 1 hour to immunoprecipitate the Arino base from the sample. (RyRl), wherein the RyR antibody is located in a modified RIPA buffer (50 mM Tris-HCl (pH 7.4), 0.9% (Nas., 5.0 mM NaF, 1.0 mM Na3 V04, 〇·5% Triton-ΧΙΟΟ and In the protease inhibitor), the sample was incubated with Protein A Sepharose microbeads (Ammersham Pharmacia Biotech, Piscatawy, NJ) for 1 hour at 4 ° C, after which the beads were washed three times with RIPA. The sample was heated to 95 ° C. And size by PAGE. Analysis: Heat the sample (immunoprecipitate or 10 pg of heart sarcoplasm,) to 95 ° C and on 6% SDS PAGE (for R γ R) and 15 % PAGE (for calstabin) Proteins were differentiated by size. Immunological spots were developed using RyR antibodies (5029, dilution ratio 1:5〇〇〇), protein kinase A phosphorylated RyR antibody (1:100〇〇) or calstabiri antibody (1:2000). The antibody was diluted in TBS-T 5% milk. 198 200815381 Reference list =

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Zhang, S.-J·, J. D. Bruton, et al. (2006)·丨, Limited oxygen diffusion accelerates fatigue development in mouse skeletal muscle. MJ Physiol (Lond) 572(2): 551-559. Brief Description of the Formulas Figures 1A and IB show the tracking data of the vehicle-treated (PBS)-treated mouse control group and the S3 6-treated mice on the first day of exercise. Figure 1A shows the speed of each mouse at five minutes and Figure 1B shows the average speed of each treatment group (η = 4 P B S, η = 4 S 3 6). There was no significant difference between treatment groups after one day of exercise. Fg-l〇〇, fg-l〇3, fg-105, and fg-i〇6 represent mice treated with S36. Fg-1〇1, fg_1〇2, fg-1〇4, and fg l〇7 represent mice treated with P B S. Figure 2A-2F shows no significant differences between the different treatment groups after 7 days of exercise. Figures 2A and 2B show the results of swimming in PBS and S36 treated mice. Figure 2A shows the speed of the five-minute interval for each mouse and Figure 2B shows the average speed for each treatment group (n = 3 PBS, n = 4 219 200815381 S3 6). Figure 2C-2F shows the results of the 7th day of the treadmill running. As described, the mice ran on a treadmill with an exercise program that increased in intensity (shown at the speed of the gray mark on the left (m/min)). Figure 2C shows individual traces showing the number of trips to the taut zone at the end of the treadmill every three minute interval. A rapid increase in the number of arrivals in the scare area (because of a mouse decline) clearly indicates that the task failed. Figure 2D shows the number of frights transmitted to each mouse every three minutes (the axis is reversed), and the mice were digitally labeled with three numerically moving averages (m 〇 v i n g average). Figure 2E depicts the total number of running distances (meters) before the failure of the PB S and S 3 6 treatment groups (η = 3 PBS, η = 4 S36), while Figure 2F depicts the number of fatigue times in the PBS and S36 treatment groups. , defined as the time the task failed (n = 3 PBS, η = 4 S3 6). No significant differences were found between the different treatment groups. Figures 3A-3F show that there is no significant difference in swimming speed between the different treatment groups after 14 days of swimming. Figure 3 shows the speed of each mouse and the third panel shows the average speed of each treatment group (n = 3 PBS, η = 4 S36). The 14th day of treadmill running showed improved performance biased towards the trend of S 3 6 treated mice. Use the increased intensity motion scheme to display the speed (m/min) marked on the left side of the gray. Figure 3C shows the various traces showing the number of trips to the tail scared area every three minutes. Figure 3D shows the number of scares (inverted) transmitted to each mouse every three minute intervals, and each mouse was plotted as a number obtained by three numerically moving average interpolation. Figure 3 depicts the total running distance (m) before failure of the PBS vs. S 3 6 treatment group (η = 3 PBS, η = 4 S36), while Figure 3F depicts the PBS vs. S36 treatment group fatigue 220 200815381 labor time The number of times, defined as the time of task failure (η = 3 PBS, η = 4 S36) 〇 The 4th, 4th, 4th and 4D graphs show the data analysis of the 16th day of the exercise process. Figures 4A and 4B show the results of the 16th day of treadmill running, which reproduces the improved performance towards the trend of s 3 6 treated mice. Use the motion scheme with increased intensity, displayed at the speed (m/min) of the black mark on the left side. Figure 4A shows the individual traces showing the number of trips to the terror zone at the end of the treadmill every three minute interval. Figure 4B shows the transmission to each mouse every three minute interval (the number of scares (the axis is reversed), and the numbers obtained by three digital moving average interpolations plot each mouse. Figure 4C depicts PBS and S36 treatment The total number of running distances (m) before the group failed (n = 3 PBS, η = 4 S3 6). Figure 4D depicts the number of fatigue times for the PBS and S36 treatment groups, defined as the time of task failure (n = 3) PBS, η = 4 S3 6). Figure 4 shows that the improved performance trend in S3 6 treated mice persisted at 16 days. Figure 5 shows S 3 6 treated mice versus PBS vehicle for the same environment on each day. The ratio of the running distance to the fatigue time of the treated mice (measured by the treadmill test). Figure 6 shows that in vivo treatment with S36 (ARM036) allows RyRl to recombine calstabirU even under intense prolonged exercise. Self-slurry immunoprecipitation of RyR 1 in mouse soleus after 21 days of exercise with or without subcutaneous micro-osmotic pump S36, followed by Western blot analysis of RyRl, phosphorus-antigenic specificity RyR, pS2844, and Binding to RyRi macromolecules Calstabini. 221 200815381 Figures 7C and 7D provide RyCal compounds for Sl〇7^, 丨, , I have less muscle atrophy pattern in mc/x mice when exercised with calcium-activated protease activity, data, and pointed out that RyCal is used Treatment of muscle-related diseases (eg, muscle contraction). Sections 7A and 7B intentionally leave blanks. Figures 8A - 8D show RjRi macromolecules 'genetic complexes undergoing substantial reorganization after exercise. Figure 8A) Immunoprecipitation of RyRi* | 1 and immunological dot analysis of RyR, RyRl-pS2844 and PDE4D3, and 蛀 human 夂 ~ combined with calstabinl of the receptor, showing exercise regimen (from twice a day swimming 槿甥 10%) for the indicated number of days The component of the RyRl complex in the long toe muscle (EDL). Fig. 8B) The optical density quantification of Fig. 8A, the respective values of which are relative to the RyR1 of the whole immunoprecipitation. Figure 8C) The composition of the RyRl complex in the long toe of the toe after five days of low-intensity and high-intensity exercise. Fig. 8D) Optical density measurement of Fig. 8C. In all of the examples, each RyR 1 immunoprecipitated product was sized, transferred and probed for a whole RyR i and one or more labeled variants on a 4-20% gradient polyacrylamide gel. The displayed ink collapse map is representative of three independent experiments. Figure 9A-9D shows that protein kinase A phosphorylation of RyR1 is caused by high-intensity bicycle exercise in humans and lacks calstabinl and PDE4D3. Figure 9A) Mud-slurry immunity from 100 ug human thigh sections before and after high-intensity (three hours at 57% V〇2 max) bicycles on days 1 and 3 (Fig. 9C) Immunoblot analysis of precipitated RyR 1 complex. The bicycle rider control group sat in the exercise room but did not exercise. Immune dot analysis showed that the overall RyRl, RyRl-S2844 protein kinase A wall acid 222 200815381, bound calstabinl and bound PDE4D3. Fig. 9B) and Fig. 9D) are quantitative analysis of optical density analysis of Figs. 9A and 9C, respectively. The bar graph depicts the levels of protein kinase Α phosphorylation, calstabinl and PDE4D3 in the RyRl complexes normalized by ryri in each day of control (n = 6) and exercise (n = 1 2) sections. Figures 10A-10F show that muscle-specific CallC mice have high-intensity motor deficits. Figure 10A) The failure time of a 2-month-old cal mouse in the same level as a wild-type mouse in a single-degree treadmill test. Dijon B) Individual treadmill failure time for each mouse by gender. Figure c) Cal 1, 1 / / mice lost weight. Figure 10D) The distribution of failure time and body weight does not show correlation in any of the mouse groups. Figure 1 〇 E) Baseline and plasma creatine kinase (CPK) levels after running on a single downhill centrifugal treadmill. i i f map) self-extended toe long muscle immunoprecipitation RyR1 and immune dot analysis RyR, RyRl_pS2844, PDE4D3 and calstabinl. *, ρ<0·01, Wilkerson check. Figure 11A-11F shows? ]^4〇-/- mice have motor deficits. Figure 11 shows the failure time of a 2-month-old call-/- mouse and a wild-type same litter in a single-degree treadmill test. Figure 11B) Individual treadmill failure time for each mouse. Figure 11C) Body weight of PDE4D··· mice. Figure 11D) The scatter plot of time to weight and weight did not show a correlation in any of the mouse groups. Figure 11E) Plasma creatinine kinase (CPK) content after running on the bottom line and a single downhill centrifugal treadmill. Fig. 1F) Self-extended toe long muscle immunoprecipitation RyRl 223 200815381 and immune dot analysis RyR, RyRl_pS2844, PDE4D3 and calstabinl. *, ρ < 0·01, Wilkerson check. Figures 12A-12F show pharmacological recombination of caistabin; [RyRi improves in vivo motor performance. Figure 12a) Failure time on a treadmill test on a specific number of days of the 28-day S1 07 treatment trial. Figure 12B) Day 21 Individual treadmill failure time for each mouse. Figure 12C) Force-frequency plot of the long toe muscles that were separated immediately after the 21st day of exercise and stimulated in the oxygenated muscle sink. The strength (eN) value is normalized by the cross-cutting area of the muscle. Figure 12D) The body weight of the entire trial did not show a therapeutic effect. The first UE map) Muscle specificity (^11 small mouse similar test treadmill failure time. Fig. 2p map) self-extension toe long muscle immunoprecipitation RyRl and immune dot analysis RyR, RyRl-pS2844, PDE4D3 and calstabinl. *,p < 〇 〇1, Salter kesen check. Figures 13A-13C show that long-term sl7 treatment reduces the likelihood of fatigue in isolated short flexor fibers. Figure )) The representative record of the flU〇_4 fluorescence value (AF/FO) of the carrier for the treatment of flexor short muscle fibers with a frequency of 12 Hz at a frequency of 0.5 Hz vocabulary - stimulating tonicity 3 〇〇 long weight The standardization of the overlying standard is standardized. Continue to ΗΕΡΡς μ I main at room temperature <> , 今 Today nWES 泰 泰 泰 液 布 布 布 布 布 布 布 布 布 布Figure 13B) S107 treatment of 瘆 似 似 一 Λ 肌 肌 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型 典型Figure 13C) The average peak of the strong mass of the flU0-4 罄 umbrella, Β! θ, 、, 4 camp first measurement (AF/FO) is the peak of the fatigue stimulus tl ^ ^ (n=11 carrier; n S107). *, independent sample t_test 13 224 200815381 1 4 A -1 4 B shows the RyR 1 system leakage type of the moving muscle, and the Po increase of the calcium ion in the stationary phase. Figure 14A) A typical record of RyRl channel activity at 90 nM [Ca2 + ]c^ from sedentary mice (left-staying, long-term exercise, and vehicle-treated mice) + vehicle) and mice that exercised for a long time and were treated with S 107 (right movement + S 1 〇 7). Single channel opening was plotted with an upward bias vector; horizontal strips were indicated at the beginning of the record Turn on and off (c) state. Display the corresponding channel open rate (P〇), average turn-on time (To) and turn-on frequency (Fo) on each record group, and display the average value of all experiments. Figure 14B) Sedentary mice (sit; η = 9) with long-term exercise and vehicle-treated mice (sports + vehicle; η = 9) and mice that were chronically exercised and treated with s 1 07 (sport + S107 ; η = 12) The average opening rate (left side), average opening time (middle) and opening frequency (right side) of RyRl activity. The error bar indicates the standard machine difference; *, compared with the sitting group p <0.005;#, compared with the exercise + S107 p < 0.005 〇 15A-15B shows S107 to avoid muscle damage caused by prolonged exercise Calcium activated protease activation. Figure 15A) The degree of plasma creatine kinase (CPK) activity in sedentary mice with long-term exercise and small gas with sate and without calstabin 1. Figure 15B) Measurement of the level of calpain activity in the homoplasmate of the long toe muscle using the calcin activating protease fluorescent matrix assay. *, independent sample t · test p < 〇. 〇 1, S107 relative carrier. Figures 16A-16C show the effect of motion on the composition of the RyRi complex. 225 200815381 The first meat fiber RyRl ^ The first S107 of the mouse [Main 17A-17B shows the distribution of 50% resorption time (τ) in the muscle in the presence or absence of S107 treatment. The 18A-18C plot shows f-induced acidification and lack of calstabin 1 over time in the mdx mouse model. Figure 19A-19F shows the effect of S107 on wild-type (unaffected) and cardiac tolerance, body weight, CPK and calpain levels. Figure 20 shows histological sections of wild type (unaffected) versus mdx mice treated with or without S107. Component Symbol Description] 226

Claims (1)

200815381 X. Application for a specialist: 1 · A method of treating or preventing muscle fatigue in a body, the method comprising administering to the individual a therapeutically effective amount of a compound of the formula Ia, Ib, Ic, I_d, Ie , If, Ig, Ih, Ii, Ik, 1-1' Im, In, Ι·ο or I-ρ, or the above-mentioned enantiomers, non-image isomers ( Diastereomers), tautomers, pharmaceutically acceptable salts, hydrates, solvates, complexes, metabolites or prodrugs, or any combination thereof, wherein the compound is not SI, S2, S3, S4, S5, S6, S7, S9, S11, S12, S13, S14, S19, S20, S22, S23, S2 4, S25, S26> S27' S36, S37, S38' S40, S43, S44 , S45' S46' S47, S48, S4 9, S50, S51, S52, S53, S54, S55, S56, S57, S58, S59, S60, S61, S62, S63, S64, S66, S67' S68, S69' S70, S7, S72, S73' S74' S75' S76' S77, S78, S79, S80, S81' S82, S83, S84, S85, S86, S87, S88, S89' S90, S91' S92' S93' S94~S95, S96, S97' S98, S99, S100 or JTV-519 〇2. The method of claim 1, wherein the compound is of the formula In, 1- Illustrated by the structure of hydrazine or Ip, or the above-described mirror image isomer, non-image isomer, tautomer, pharmaceutically acceptable salt, hydrate, solvate, complex, metabolite or prodrug Or any of the above-mentioned groups 227, 2008, 158, 381, and the method of claim 1, wherein the compound is selected from the group consisting of: S101, S102, S103, S104, S107, S108, S109 , S110, Sm, S112, S113, S114, S115, S116, S117, S118, S119, S120, S121, S122 and S123, and any of the above salts, hydrates, solvates, polymorphs, A complex, metabolite or prodrug, or a combination of any of the foregoing. 4. The method of any of claims 2 or 3, wherein the muscle fatigue is caused by a skeletal muscle disease or abnormality. 5. The method of claim 4, wherein the skeletal muscle disease or abnormality is associated with a functional abnormality of a type 1 anaphytic receptor (RyRl). 6. The method of claim 4, wherein the skeletal muscle disease or abnormal myopathy. 7. The method of claim 6, wherein the myopathy is a muscular dystrophy, a central core disease, or a malignant hyperthermia. The method according to any one of claims 1 to 2, wherein the muscle fatigue is caused by a disease or a symptom selected from the group consisting of: Neur〇pathy), neurological disease or abnormality, seizure condition, genetic disease or abnormality, heart disease or abnormality, infectious disease, HIV infection, AIDS, cancer fatigue, malnutrition, kidney disease and kidney failure. 9. The method of claim 8, wherein the heart disease or abnormality is selected from the group consisting of: an irregular heartbeat, an irregular heartbeat caused by exercise, congestive heart failure, chronic Chronic obstructive pulmonary disease, hypertension, or any combination of the above. The method of claim 9, wherein the irregular heartbeat symptom is selected from the group consisting of: atrial or ventricular arrhythmia or atrial fibrillation; atrial or ventricular Atrial or ventricular tachyarrhythmia; atrial or ventricular tachycardia; catecholaminergic polymorphic ventricular tachycardia (CPVT) and its motor-induced variants. 229. The method of any one of the preceding claims, wherein the muscle fatigue is caused by exercise-induced muscle fatigue. 1 2 The method of claim 11, wherein the muscle fatigue caused by the exercise is caused by prolonged exercise or high-intensity exercise. The method of claim 1, wherein the system is selected from the group consisting of a non-human animal, a canine family, a horse family, a cat family, a pig class, A method of the invention, wherein the system is a human and the compound is contained in a pharmaceutical composition, the pharmaceutical composition of the invention. The composition includes at least one pharmaceutically acceptable excipient. The method of claim 14, wherein the at least one pharmaceutically acceptable excipient is selected from the group consisting of: aromatics, buffers, adhesives (binders) ), colorants, disintegrants, thinners, emulsifiers, extenders, flavor-improving agents, gellants, slip agents (glidants), preservatives, skin-penetration enhancers, solubilizers, 230 200815381 stabilizers, suspending agents, sweeteners, tonicity The method of claim, the vehicle, the viscosity-enhancing agent, or any combination of the above, wherein the pharmaceutical composition further comprises a second active agent. . The method of claim 16, wherein the second active agent is an analgesic. 18. The method of claim 17, wherein the composition is in the form of a capsule, a granule form, a powder form, a solution form, a suspension form or a It exists in the form of a tablet. The method of claim 18, wherein the composition is administered orally or parenterally via a drug release stent or an implant or via a osmotic pump ( Parenterally), enteral, intravenous, intraarterial, subcutaneous, intramuscularly administered. 231. The method of claim 1, wherein the method is one of an activity sufficient to reduce sputum activating protease (c a 1 p a i η ) activity. The method of claim 1, wherein the method is administered to the individual in a dose sufficient to reduce plasma creatine kinase. 22. The use of a therapeutically effective dose of a compound of the formula 1- um, 1-13, 1-〇, 1-(1, 1-6, 1-£, 1-8, 1- The structure of Ik, 1-1, Ι-η, 1-〇 or Ι-ρ, or an isomer, a non-image isomer, a tautomer, a pharmaceutically, a hydrate, a solvate, a compound, a metabolite or a combination of the foregoing; for the preparation of a muscle fatigue in the body wherein the compound is not SI, S2, S3, S4, S5 S9, S11, S12, S13, S14 'S19, S20 'S24, S25 > S26 > S27, S36, S37, S38, S44 - S45 > S46 'S47, S48, S49, S50, S53, S54, S55, S56, S57, S58, S59, S62 - S63 'S64 ' S66, S67, S68, S69 'S72 > S73 > S74 > S75, S76, S77 'S78 > S81, S82 > S83, S84, S85, S86, S87, the compound is applied to the compound system Or the content of the compound h, Ii, Ij, the above image. Acceptable salt 'drug, or any composition, S6 'S7 ' S22 ' S23 ^ S40 > S43 ^ S51 , S52 , S60 ^ S6 Bu S70 ' S71 > S79 , S80-S88, S89, 232 200815381 S90, S91, S92 > S93, S94 > S95 'S96, S97 'S98, S99, S100 or JTV-519. 2 3 · A therapeutically effective dose of one of the compounds Use, wherein the compound is represented by the structure of Ι-ll, 1-〇 or Ip, or the above-mentioned mirror image isomer, non-image isomer, tautomer, pharmaceutically acceptable salt, a hydrate, solvate, complex, metabolite or prodrug, or a combination of any of the foregoing; for the preparation of a composition for treating an in vivo muscle fatigue. 24. If the patent application is in claim 22 or 23 The use, wherein the compound is selected from the group consisting of: S 1 0 1 , S 1 0 2, S103, S104, S107, S108, S109, S110, Sill, S112, S113, S114, S115, S116, S117, S118, S119, S120, S121, S122 and S 1 2 3 'and any of the salts, hydrates, solvates, polymorphs, complexes, metabolites or prodrugs thereof, and any of the foregoing combination. 233