Creatine prodrug, its composition, and method of use thereof

Abstract

This invention provides a membrane-permeable creatine prodrug in which the creatine molecule is modified to be more stable and to penetrate barrier tissues and cell membranes more effectively. [Solution] The present invention provides a membrane-permeable creatine prodrug, a pharmaceutical composition containing the prodrug, and a method for treating diseases such as ischemia, heart failure, neurodegenerative diseases, and genetic disorders affecting the creatine kinase system, comprising administering the prodrug or the pharmaceutical composition thereof. The creatine prodrug is, for example, a compound of formula (VII). JPEG2026048846000136.jpg61170

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Application No. 62 / 095,295, filed December 22, 2014, titled "CREATINE PRODRUGS, COMPOSITIONS AND METHODS OF USE THEREOF," the disclosure of which is incorporated herein by reference in its entirety for all purposes.

[0002] Field of Invention The present invention describes membrane-permeable creatine prodrugs, pharmaceutical compositions comprising membrane-permeable creatine prodrugs, and methods for treating diseases, such as ischemia, heart failure, neurodegenerative diseases, and genetic disorders affecting the creatine kinase system, comprising administering a creatine prodrug or a pharmaceutical composition thereof. In some embodiments, the present invention describes the treatment of genetic diseases affecting the creatine kinase system, such as creatine transporter disorders or creatine synthesis disorders, comprising administering a creatine prodrug or a pharmaceutical composition thereof. [Background technology]

[0003] Creatine plays a crucial role in cellular energy metabolism, and as high-energy phosphocreatine, it constitutes a significant muscle energy store in addition to adenosine triphosphate (ATP). In resting muscles, ATP can form phosphocreatine by transferring phosphate groups to creatine, and thus phosphocreatine is in direct equilibrium with ATP. While muscles are working, phosphocreatine is critical for the fastest possible replenishment of ATP stores. Phosphocreatine is available for this purpose during the first few seconds of maximum muscle load; this substance can reconstruct ATP in a very rapid reaction by the enzyme creatine kinase, which transfers phosphate groups to adenosine diphosphate. The creatine kinase system has a dual role in intracellular energy metabolism and function: as an energy buffer to restore ATP levels depleted at high-ATP hydrolysis sites, and as an energy buffer to transfer energy from mitochondria to other cellular parts in the form of phosphocreatine through processes involving intermediate energy carriers, multiple enzymatic reactions, and diffusion through various intracellular structures.

[0004] Many pathological conditions arise from dysfunction of energy metabolism. Depletion of ATP stores in cells, for example, when occurring during tissue ischemia, leads to impaired tissue function and cell death. Among the most medically relevant are ischemia-related cardiovascular diseases, such as stroke and heart attack, which remain the leading causes of death and morbidity in North America and Europe. Therefore, strategies that can prevent or reverse ischemia-related tissue damage are expected to have a significant impact on public health. Energy depletion is also a contributing factor to tissue damage during surgery and is a common cause of organ transplant failure. Furthermore, reperfusion with oxygen-containing solutions can even worsen tissue integrity through the generation of oxygen radicals. Thus, methods to rapidly restore ATP levels without causing reperfusion injury appear to have many therapeutic applications. Neurodegenerative diseases, such as Parkinson's disease, Alzheimer's disease, and Huntington's disease, are associated with energy metabolism disorders, and strategies to improve ATP metabolism are thought to minimize neuronal loss and thereby improve the prognosis of patients with these diseases. Finally, impaired energy metabolism is a major factor in muscle fatigue and limits physical endurance. Therefore, methods to prevent or reverse ATP depletion in ischemic or metabolically active tissues appear to have broad clinical utility for a wide range of applications.

[0005] Creatine supplementation increases intracellular creatine phosphate levels (Harris et al.) (Dechent et al., Clinical Sci 1992, 83, 367-74). In healthy individuals, creatine readily crosses the blood-brain barrier, and therefore, oral administration can increase cerebral creatine levels (Dechent et al., Am J Physiol 1999, 277, R698-704). Long-term creatine supplementation can increase the pooling of creatine phosphate in cells, thereby enhancing resistance to tissue ischemia and muscle fatigue. In other words, while creatine administration may have some therapeutic efficacy, modified creatine molecules that are more stable and more permeable to barrier tissues and cell membranes are likely to have even greater therapeutic value.

[0006] The creatine prodrugs of the present invention are stable in biological fluids and are designed to enter cells by either passive diffusion or active transport, and to release creatine into the cytoplasm of cells. These prodrugs can also cross important barrier tissues, such as the intestinal mucosa, the blood-brain barrier, and the blood-placental barrier. Thanks to their ability to cross biological membranes, creatine prodrugs can restore and maintain energy homeostasis in ATP-depleted cells via the creatine kinase system, and rapidly restore ATP levels to protect tissues from further ischemic stress. Creatine prodrugs with higher free energy or lower creatine kinase affinity that can regenerate ATP under more severe energy depletion conditions are also disclosed. The creatine prodrugs of the present invention can also be used for sustained systemic delivery of creatine at certain concentrations. The present invention relates to these, as well as other important objectives. [Overview of the project]

[0007] The present invention relates to a membrane-permeable creatine prodrug, a pharmaceutical composition containing the membrane-permeable creatine prodrug, and a method of using the membrane-permeable creatine prodrug and its pharmaceutical composition. In some embodiments, the present invention describes the treatment of genetic diseases affecting the creatine kinase system, such as creatine transporter disorders or creatine synthesis disorders, including administering a creatine prodrug or its pharmaceutical composition.

[0008] In one embodiment, the present invention describes a compound of formula (I), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (I) is as follows JPEG2026048846000001.jpg72128 Where: R is -CH3 or -CD3; R 1 is hydrogen, -OR 2 , -C(O)OR 2 , -C(O)R 2 , TIFF2026048846000002.tif2671 , TIFF2026048846000003.tif2657 , JPEG2026048846000004.jpg2737 , or JPEG2026048846000005.jpg1752 ; n is an integer from 1 to 2; Each R 2 is independently hydrogen, C 1-12 alkyl, substituted C 1-12 alkyl, C 1-12 heteroalkyl, substituted C 1-12 heteroalkyl, C 3-12 cycloalkyl, substituted C 3-12 cycloalkyl, C4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl; Each R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl; R 23 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 5-12 Cycloalkyl, substituted C 5-12 Cycloalkyl, C 5-12 Aryl, and C 5-12 Substitutive aryl, -C(O)-OR 22 , or -C(O)-R 22 and; R 22 C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl , replace C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl; and R 48 C 1-12 Alkyl or substituted C 1-12 It is alkyl.

[0009] Further embodiments describe the compound of formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (III) is as follows: JPEG2026048846000006.jpg41128 During the ceremony: W is -CH2OH or -C(O)OR 7 and; R is either -CH3 or -CD3; R 7 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl, substituted C 6-20 Heteroarylalkyl, -C(O)R 5 , -C(O)OR 5 -C(O)(NR 3 R4 )、 -C(R 3 R 4 )-C(O)OR 22 、 -C(R 3 R 4 )-(O)C(O)R 22 、 -C(R 3 R 4 )-(O)C(O)-OR 22 、 TIFF2026048846000007.tif3260 、 TIFF2026048846000008.tif3245 、 or JPEG2026048846000009.jpg2323 <00​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​23 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 5-12 Cycloalkyl, substituted C 5-12 Cycloalkyl, C 5-12 Aryl, and C 5-12 Substitutive aryl, -C(O)-OR 22 , or -C(O)-R 22 and R 22 C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl group.

[0010] Further embodiments describe the compound of formula (VI), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (VI) is as follows: JPEG2026048846000010.jpg37128 During the ceremony: R is either -CH3 or -CD3; R 10 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl, substituted C 6-20 Heteroarylalkyl, -C(O)R 5 , -C(O)OR 5 -C(O)(NR 3 R 4 ), -C(R 3 R 4 )-C(O)OR 22 , -C(R 3 R 4 )-(O)C(O)R 22 , -C(R 3 R 4 )-(O)C(O)-OR 22 ; JPEG2026048846000011.jpg2323 , TIFF2026048846000012.tif2659 ,or TIFF2026048846000013.tif2644 and; R 11 and R 12 These are, independently, hydrogen or -OR 13 is it; or R 11 and R 12 These are -C(O)R respectively. 5 However, R 11 and R12 Both cannot be hydrogen; R 13 These are, independently, hydrogen and C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl, substituted C 6-20 Heteroarylalkyl-CH(OR 5 ), -C(O)R 5 , -C(O)OR 5 , or -C(O)(NR 3 R 4 ) and; Each R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl; R 5 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl; R 23 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 5-12 Cycloalkyl, substituted C 5-12 Cycloalkyl, C 5-12 Aryl, and C 5-12 Substitutive aryl, -C(O)-OR 22 , or -C(O)-R 22 and; R 22 C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl; and n is an integer between 1 and 3.

[0011] One other embodiment describes a compound of formula (VII), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (VII) is as follows: JPEG2026048846000014.jpg39128 During the ceremony: R is either -CH3 or -CD3; Each R 14 These are, independently, hydrogen and C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substitution C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl, substituted C 6-20 Heteroarylalkyl-CH(OR 5 ), -C(O)R 5 , -C(O)OR 5 , or -C(O)(NR 3 R 4 ) and; Each R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 alkyl and R 5 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl group.

[0012] In certain embodiments, compounds of formulas (I), (III), (VI), and (VII) may have the following properties:

[0013] Each R is independently -CH3.

[0014] Each R is independently -CD3.

[0015] Each n is an independent integer of 1.

[0016] Each n is an independent integer 2.

[0017] Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 14 , R 15 , R 18 , and R 22 Independently, C 1-6 Alkyl, substituted C 1-6 Alkyl, C 3-7 Cycloalkyl, substituted C 3-7 Cycloalkyl, C 5-7 Aryl, or substitution C 5-7 It is Ariel.

[0018] Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 14 , R 15 , R 18 , and R 22 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-dimethoxyethyl, 1,1-diethoxyethyl, phenyl, 4-methoxyphenyl, benzyl, phenethyl, styryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl.

[0019] Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 14 , R 15 , R 18 , and R 22 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-diethoxyethyl, phenyl, cyclohexyl, or 3-pyridyl.

[0020] Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 14 , R 15 , R 18 , and R 22 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, dodecyl, tert-butyl, phenyl, or cyclohexyl.

[0021] Each R 2 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 14 , R 15 , R 18 , and R 22 These are independently ethyl, isopropyl, or dodecyl.

[0022] Each R 3 and R 4 It is, independently, hydrogen.

[0023] Each R 23 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl.

[0024] Each R 23 It is methyl.

[0025] Each substituent is independently halogen, -NO2, -OH, -NH2, -CN, -CF3, -OCF3, =O, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Alkoxy or substituted C 1-12 Alkoxy, -COOR 10’ And in the formula, R 10’ is hydrogen, C 1-3 Alkyl, or -(NR) 11’ )2, and in the formula, each R 11’ These are, independently, hydrogen or C 1-3 It is alkyl.

[0026] In one embodiment, the compound of formula (I) is a compound of formula (X), formula (XI), formula (XII), formula (XIII), formula (XIV), formula (XV), formula (XVa), or formula (XVb), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof; The compound of formula (X) is as follows: JPEG2026048846000015.jpg74128 In the formula, R is either -CH3 or -CD3; The compound of formula (XI) is as follows: JPEG2026048846000016.jpg73128 In the formula, R is -CH3 or -CD3; and R 24 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; The compound of formula (XII) is as follows: JPEG2026048846000017.jpg74128 In the formula, R is -CH3 or -CD3; and R 25 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; The compound of formula (XIII) is as follows: JPEG2026048846000018.jpg73128 In the formula, R is -CH3 or -CD3; and R 26 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; The compound of formula (XIV) is as follows: JPEG2026048846000019.jpg31128 The compound of formula (XV) is as follows: JPEG2026048846000020.jpg32128 In the formula, R is either -CH3 or -CD3; The compound of formula (XVa) is as follows: JPEG2026048846000021.jpg86170 In the formula, R is either -CH3 or -CD3; R 39 is hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; and R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl; The compound of formula (XVb) is as follows: JPEG2026048846000022.jpg73170 In the formula, R is either -CH3 or -CD3; R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 alkyl and R 53 C 1-12 Alkyl or substituted C 1-12 It is alkyl.

[0027] In yet another embodiment, the compound of formula (III) is a compound of formula (XVII), formula (XVIII), or formula (XIX), or a pharmaceutically acceptable salt, solvate thereof. It is a variant or stereoisomer; The compound of formula (XVII) is as follows: JPEG2026048846000023.jpg40128 In the formula, R is either -CH3 or -CD3; R 29The compounds are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, -cyclohexyl, -CH2-C(O)OR 43 -CH2-(O)C(O)R 43 -CH2-(O)C(O)OR 43 ,or JPEG2026048846000024.jpg2522 and; R 39 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; R 43 is hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; and R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl; The compound of formula (XVIII) is as follows: JPEG2026048846000025.jpg33128 In the formula, R is either -CH3 or -CD3; The compound of formula (XIX) is as follows: JPEG2026048846000026.jpg34128 In the formula, R is either -CH3 or -CD3.

[0028] In yet another embodiment, the compound of formula (VI) is a compound of formula (XXII), formula (XXIII), formula (XXIV), formula (XXV), formula (XXVI), formula (XXVII), or formula (XXVIII), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof; The compound of formula (XXII) is as follows: JPEG2026048846000027.jpg36128 The compound of formula (XXIII) is as follows: JPEG2026048846000028.jpg36128 The compound of formula (XXIV) is as follows: JPEG2026048846000029.jpg36128 The compound of formula (XXV) is as follows: JPEG2026048846000030.jpg32128 The compound of formula (XXVI) is as follows: JPEG2026048846000031.jpg37128 The compound of formula (XXVII) is as follows: JPEG2026048846000032.jpg36128 The compound of formula (XXVIII) is as follows: JPEG2026048846000033.jpg42128 In the formula, R is either -CH3 or -CD3; R a These are hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl; R 32 The compounds are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, cyclohexyl, -CH2-C(O)OR 43 -CH2-(O)C(O)R 43 -CH2-(O)C(O)OR 43 ,or JPEG2026048846000034.jpg2522 and; R 39 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; R 33 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; R 43 is hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; and R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl.

[0029] In yet another embodiment, the compound of formula (VII) is the compound of formula (XXIX), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof; The compound of formula (XXIX) is as follows: JPEG2026048846000035.jpg37128 In the formula, R is -CH3 or -CD3; and Each R 34 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl.

[0030] In one embodiment, the present invention relates to a compound having the following structure: TIFF2026048846000036.tif1333 , TIFF2026048846000037.tif1433 , TIFF2026048846000038.tif1329 、 TIFF2026048846000039.tif1429 、 TIFF2026048846000040.tif1330 、 TIFF2026048846000041.tif1430 、 TIFF2026048846000042.tif1333 TIFF2026048846000043.tif1433 、 TIFF2026048846000044.tif1329 、 TIFF2026048846000045.tif1429 、 TIFF2026048846000046.tif1351 、 TIFF2026048846000047.tif1451 、 TIFF2026048846000048.tif1419 、 TIFF2026048846000049.tif1919 、 TIFF2026048846000050.tif1420 、 TIFF2026048846000051.tif1920 、 TIFF2026048846000052.tif1533 、 TIFF2026048846000053.tif1533 , TIFF2026048846000054.tif1836 , TIFF2026048846000055.tif1836 , TIFF2026048846000056.tif2131 , TIFF2026048846000057.tif2131 , TIFF2026048846000058.tif2125 ,or TIFF2026048846000059.tif2225 ; or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof. Write it down.

[0031] In another embodiment, the present invention describes a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I), (III), (VI), and (VII), and at least one of any subtype or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or a pharmaceutically acceptable solvate of any of the above, and a pharmaceutically acceptable vehicle. In one embodiment, the present invention describes a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds as disclosed herein, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or a pharmaceutically acceptable solvate of any of the above, and a pharmaceutically acceptable vehicle.

[0032] Depending on the embodiment, the pharmaceutical composition may be incorporated into one or more sustained-release oral dosage forms.

[0033] In one embodiment, the pharmaceutical composition contains at least one compound of the present invention in an amount effective for treating a patient's disease, which is ischemia, oxidative stress, neurodegenerative disease, ischemia-reperfusion injury, cardiovascular disease, genetic diseases affecting the creatine kinase system, multiple sclerosis, mental disorders, and muscle fatigue; or contains in an amount sufficient to bring energy homeostasis to the diseased tissue or organ; or contains in an amount effective for improving the patient's muscle strength; or contains in an amount effective for improving the viability of the tissue or organ; or contains in an amount effective for improving the viability of the cell. In another embodiment, the pharmaceutical composition contains at least one compound of the present invention in an amount effective for treating a genetic disease affecting the creatine kinase system. In some embodiments, the pharmaceutical composition contains at least one compound of the present invention in an amount effective for treating creatine transporter disorders. In one embodiment, the pharmaceutical composition contains at least one compound of the present invention in an amount effective for treating creatine synthesis disorders.

[0034] In one embodiment, the present invention describes a method for treating diseases associated with impaired energy metabolism in patients, such as ischemia, oxidative stress, neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), Huntington's disease, Parkinson's disease, or Alzheimer's disease, ischemia-reperfusion injury, cardiovascular disease, multiple sclerosis (MS), psychiatric disorders, genetic diseases affecting the creatine kinase system, or muscle fatigue, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof.

[0035] In another embodiment, a method is described for treating a genetic disorder affecting a patient's creatine kinase system, such as a creatine transporter disorder or a creatine synthesis disorder, the method comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof.

[0036] In a further embodiment, a method for improving a patient's muscle strength is described, which involves administering to a patient in need of such improvement, in a therapeutically effective amount, a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof.

[0037] In yet another embodiment, a method for increasing the viability of a tissue or organ is described, wherein the tissue or organ is treated with an effective amount of at least one of the compounds of formula (I), (III), (VI), (VII), and any subtype or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or the compounds of formula (I), (III), (VI), (VII), and any subtype or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof. This includes contacting a pharmaceutical composition containing a substance.

[0038] In another embodiment, a method for improving the viability of isolated cells is described, which involves contacting cells with an effective amount of a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or with a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof.

[0039] In another embodiment, a method for treating a disease associated with oxidative stress is described, comprising administering to a patient in need of such treatment, in an effective amount, a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof.

[0040] In another embodiment, a method is described for improving the viability of a tissue or organ to treat a tissue or organ exhibiting impaired energy metabolism, the method comprising contacting a tissue or organ with a compound of formula (I), (III), (VI), (VII), and at least one of any subspecies or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or with a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subspecies or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof.

[0041] In yet another embodiment, a method for inducing energy homeostasis in a tissue or organ is described, which comprises contacting a tissue or organ with a compound of formula (I), (III), (VI), (VII), and at least one of any subspecies or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or with a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subspecies or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof.

[0042] In another embodiment, a method for treating oxidatively stressed tissue or organ is described, which involves contacting the tissue or organ with a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof, or with a pharmaceutical composition comprising a compound of formula (I), (III), (VI), (VII), and at least one of any subtypes or chemical species thereof, or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the invention pertains. In the specification, singular nouns also include plural nouns unless the context explicitly states otherwise. Methods and materials similar to or equivalent to those described herein may be used in carrying out or testing the invention, but suitable methods and materials are described below. All publications, patent applications, patents, and other references referred to herein are incorporated by reference. References made herein are not considered prior art of the claimed invention. In the event of any inconsistency, this specification shall not be deemed to be prior art. The regulations also apply to the meaning of the matter. Furthermore, the materials, methods, and examples are illustrative only and not intended to be restrictive.

[0044] Other features and advantages of the present invention will become apparent from the following detailed description and claims. [Modes for carrying out the invention]

[0045] definition A dash ("-") that is not between two letters or symbols is used to indicate a bond point of a part or substituent. For example, -CONH2 is bonded through a carbon atom.

[0046] "Alkyl" refers to a saturated or unsaturated, branched or linear, monovalent hydrocarbon radical, derived by removing one hydrogen atom from one carbon atom of a parent alkane, alkene, or alkyne, either by itself or as part of another substituent. Examples of alkyl groups include methyl; ethyl groups, e.g., ethanyl, ethenyl, and ethynyl; propyl groups, e.g., propan-1-yl, propan-2-yl, prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl(allyl), prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; and butyl groups, e.g., butyl. n-1-yl, butan-2-yl, 2-methyl-propan-1-yl, 2-methyl-propan-2-yl, but-1-en-1-yl, but-1-en-2- Examples include but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, but-1,3-dien-1-yl, but-1,3-dien-2-yl, but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc., but are not limited to these. It is not determined.

[0047] The term "alkyl" specifically includes groups having any degree or level of saturation, i.e., groups having only carbon-carbon single bonds, groups having one or more carbon-carbon double bonds, groups having one or more carbon-carbon triple bonds, and groups having a mixture of carbon-carbon single, double, and triple bonds. The terms "alkanyl," "alkenyl," and "alkynyl" are used when referring to a specific level of saturation. Alkyl groups may have 1 to 20 carbon atoms in certain embodiments, 1 to 12 carbon atoms in certain embodiments, 1 to 10 carbon atoms in certain embodiments, 1 to 6 carbon atoms in certain embodiments, and 1 to 3 carbon atoms in certain embodiments.

[0048] "Alkoxy" can be radical OR, either by itself or as part of another substituent. 31 This shows that in the formula, R 31 The group is selected from alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, cycloalkylalkyl, heterocycloalkylalkyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl as defined herein. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, and cyclohexyloxy.

[0049] "Aryl" refers to a monovalent aromatic hydrocarbon radical derived from the removal of one hydrogen atom from one carbon atom in an aromatic ring system, either by itself or as part of another substituent. Aryls include five- and six-membered carbocyclic aromatic rings, e.g., benzene; bicyclic ring systems in which at least one ring is carbocyclic and aromatic, e.g., naphthalene, indan, and tetralin; and tricyclic ring systems in which at least one ring is carbocyclic and aromatic, e.g., fluorescein. Aryls encompass a multi-ring system having at least one carbocyclic aromatic ring and at least one carbocyclic aromatic ring, cycloalkyl ring, or heterocycloalkyl ring fused with it. For example, aryls include 5- and 6-membered carbocyclic aromatic rings fused with 5- to 7-membered heterocycloalkyl rings containing one or more heteroatoms selected from N, O, and S. In such a fused bicyclic ring system where only one of the rings is a carbocyclic aromatic ring, the bond site may be on the carbocyclic aromatic ring or on the heterocycloalkyl ring. Examples of aryl groups include, but are not limited to, groups derived from acetantrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, coronene, fluorantene, fluorene, hexacene, hexaphene, hexylene, as-indacene, s-indacene, indan, indene, naphthalene, octacene, octafen, octaene, ovalene, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiaden, pyrene, pyranthrene, rubicene, triphenylene, trinaphthalene, and others. In certain embodiments, an aryl group may have 6 to 20 carbon atoms, 6 to 12 carbon atoms, and in certain embodiments, 6 to 8 carbon atoms. However, aryls do not in any way encompass or overlap with heteroaryls as separately defined herein.

[0050] "Arylalkyl" refers to a carbon atom, typically terminal or sp, either by itself or as part of another substituent. 3This represents an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom is replaced by an aryl group. Examples of arylalkyl groups include, but are not limited to, benzyl, 2-phenylethane-1-yl, 2-phenylethen-1-yl, naphthylmethyl, 2-naphthylethane-1-yl, 2-naphthylethen-1-yl, naphthobenzyl, and 2-naphthophenylethane-1-yl. When a specific alkyl moiety is intended, the nomenclature arylalkanyl, arylalkenyl, or arylalkynyl is used. In certain embodiments, the arylalkyl group is C 6-30 The arylalkyl group, for example, the alkanyl, alkenyl, or alkynyl portion of the arylalkyl group, is C 1-10 The aryl portion is C 6-20 In a particular embodiment, the arylalkyl group is C 6-20 The arylalkyl group, for example, the alkanyl, alkenyl, or alkynyl portion of the arylalkyl group, is C 1-8 The aryl portion is C 6-12 It belongs to them.

[0051] AUC is the area under the curve representing the concentration of a compound or its metabolites in a patient's bodily fluids as a function of time after administration of the compound to the patient. In certain embodiments, the compound can be a prodrug, and the metabolite can be a drug. Examples of bodily fluids include plasma and blood. AUC may be determined by measuring the concentration of a compound or its metabolites in bodily fluids, such as plasma or blood, at various time intervals using methods such as liquid chromatography-tandem mass spectrometry (LC / MS / MS), and calculating the area under the plasma concentration-time curve. Appropriate methods for calculating AUC from drug concentration-time curves are well known in the art. Where applicable to the present invention, the AUC of a drug or its metabolites may be determined by measuring the drug concentration over time in the patient's plasma, blood, or other bodily fluids or tissues after administration of the compound of the present invention to the patient.

[0052] "Bioavailability" refers to the rate and amount of a drug or its prodrug that reaches the patient's systemic circulation after administration. This can be determined, for example, by evaluating the plasma or blood concentration-over-time characteristics of the drug. Useful parameters for characterizing the plasma or blood concentration-over-time curve include the area under the curve (AUC) and the time to peak concentration (T). max ), and the maximum drug concentration (C max ) are listed, C max T is the highest concentration of the drug in a patient's plasma or blood after administration of one dose or one dosage form of the drug to the patient. max This refers to the plasma or blood of a patient after administration of one dose or one dosage form of the drug to the patient. The drug reaches its highest concentration (C max This is the time it takes to reach ).

[0053] "C max " is the highest concentration of the drug in the patient's plasma or blood after administration of one dose of the drug or prodrug to the patient.

[0054] "T max "This refers to the point at which the drug reaches its peak concentration (C) in the patient's plasma or blood after administration of one dose of the drug or prodrug to the patient. max This is the time it takes to reach ).

[0055] "The compound(s) of the present invention" or "the compound of the present invention" encompasses any of the specific compounds in these formulas. Compounds may be identified by either their chemical structure and / or chemical name. If the chemical structure and / or chemical name are inconsistent, the chemical structure is the determining factor in identifying the compound. Compounds described herein may have one or more chiral centers and / or double bonds and therefore may exist as stereoisomers, such as double bond isomers (i.e., geometric isomers), enantiomers, or diastereomers. Accordingly, within the scope of the specification, any chemical structure described in whole or in part with its relative configuration encompasses all possible enantiomers and stereoisomers of the exemplified compound, including their pure form as stereoisomers (e.g., geometrically pure, pure as an enantiomer, or pure as a diastereomer) as well as enantiomer mixtures and stereoisomer mixtures. Enantiomer mixtures and stereoisomer mixtures can be separated into their constituent enantiomers or stereoisomers using separation techniques or chiral synthesis techniques well known to those skilled in the art. The compounds of the present invention are also referred to as "creatine prodrugs" or "prodrugs of the present invention."

[0056] Examples of compounds of the present invention include, but are not limited to, stereoisomers or optical isomers of the compounds of the present invention, their racemates, and other mixtures thereof. In such embodiments, a single enantiomer or diastereomer, i.e., an optically active form, can be obtained by asymmetric synthesis or by resolution of a racemate. Resolution of a racemate can be achieved, for example, by crystallization in the presence of a resolving agent, or by conventional methods such as chromatography using a chiral high-pressure liquid chromatography (HPLC) column. The compounds of the present invention also include Z- and E-forms (or cis- and trans-forms) of compounds having a double bond. In embodiments in which the compounds of the present invention exist in various tautomer forms, the compound includes all tautomer forms of that compound.

[0057] A "stereoisomer" refers to a compound that consists of the same atoms and the same bonds, but has a different three-dimensional structure, and these structures are not interconvertible. This invention aims to encompass various stereoisomers and mixtures thereof, and includes "enantiomers." Enantiomers refer to two stereoisomers whose molecules are mirror images of each other and do not overlap.

[0058] The compounds of the present invention may exist in multiple tautomers, and when only one tautomer is described herein, it is for convenience only and naturally includes other tautomers in the shown form. Therefore, the chemical structures depicted herein encompass all possible tautomers of the exemplified compounds. The term "tautomer," as used herein, refers to isomers that can coexist in equilibrium because they readily transform into each other. For example, ketones and enols are two tautomers of a single compound. In another example, substituted 1,2,4-triazole derivatives may exist in at least three tautomers, as shown below: JPEG2026048846000060.jpg2586 R T1 is an alkyl group which may have H or substituents, R T2 This is an aryl which may have substituents.

[0059] The compounds of the present invention also include isotope-labeled compounds in which one or more atoms have atomic masses different from those normally found in nature. Examples of isotopes that can be incorporated into the compounds disclosed herein include: 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18 O, 17Examples include, but are not limited to, O. Compounds may exist in non-solvated forms, solvated forms including hydrated forms, and as N-oxides. Generally, compounds can be hydrated, solvated, or N-oxides. Certain compounds may exist in multiple crystalline or amorphous forms. The compounds of the present invention include pharmaceutically acceptable salts thereof or pharmaceutically acceptable solvates of any of the above free acid forms, as well as any of the above crystalline forms.

[0060] The "creatine kinase system" includes, but is not limited to, creatine transporters, creatine, creatine kinase, creatine phosphate, and intracellular energy transport of creatine, creatine kinase, and / or creatine phosphate. Examples of creatine kinase systems include the mitochondrial creatine kinase system and the cytoplasmic creatine kinase system. Actions related to the creatine kinase system refer to the transport, synthesis, metabolism, and translocation of compounds and proteins included in the creatine kinase system.

[0061] "Cycloalkyl" refers to a saturated or partially unsaturated cyclic alkyl radical, either by itself or as part of another substituent. The nomenclature "cycloalkanyl" or "cycloalkenyl" is used when a specific level of saturation is intended. Examples of cycloalkyl groups include, but are not limited to, groups derived from cyclopropane, cyclobutane, cyclopentane, cyclohexane, and others. In certain embodiments, the cycloalkyl group is C 3-15 Cycloalkyl, C 5-12 It is a cycloalkyl, and in a particular embodiment, C 3-7 It is a cycloalkyl group.

[0062] "Cycloalkylalkyl" refers to a carbon atom, typically terminal or sp, either by itself or as part of another substituent. 3This represents an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom is replaced by a cycloalkyl group. When a specific alkyl moiety is intended, the nomenclature cycloalkylalkanyl, cycloalkylalkenyl, or cycloalkylalkynyl is used. In certain embodiments, the cycloalkylalkyl group is C 7-30 Cycloalkylalkyl, for example, the alkanyl, alkenyl, or alkynyl portion of a cycloalkylalkyl group is C 1-10 The cycloalkyl portion is C 6-20 In some embodiments, the cycloalkylalkyl group is C 7-20 Cycloalkylalkyl, for example, the alkanyl, alkenyl, or alkynyl portion of a cycloalkylalkyl group is C 1-8 The cycloalkyl portion is C 4-20 or C 6-12 It is so.

[0063] "Disease" refers to a disease, disorder, symptom, sign, or indication.

[0064] "Halogen" refers to fluoro, chloro, bromo, or iodo groups.

[0065] "Heteroalkyl" refers to an alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) are independently replaced by the same or different heteroatomic group, either by themselves or as part of another substituent. Examples of heteroatomic groups include -O-, -S-, -OO-, -SS-, -OS-, and -NR. 57 R 58 -, =NN=, -N=N-, -N=N-NR 59 R 60 ,-PR 61 -, -P(O)2-, -POR 62 -, -OP(O)2-, -SO-, -SO2-, -SnR 63 R 64 - are examples, but are not limited to these, and in the formula, R 57 , R 58 , R 59 , R 60, R 61 , R 62 , R 63 , and R 64 These are, independently, hydrogen and C 1-12 Alkyl, substituted C 1-12 Alkyl, C 6-12 Aryl substitution C 6-12 Ariel, C 7-18 Arylalkyl, substituted C 7-18 Arylalkyl, C 3-7 Cycloalkyl, substituted C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, substituted C 3-7 Heterocycloalkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 6-12 Heteroaryl, substituted C 6-12 Heteroaryl, C 7-18 Heteroarylalkyl or substituted C 7-18 Selected from heteroarylalkyls. If a specific level of saturation is intended, "heteroalkanyl", "heteroalkenyl", or R 60 , R 61 , R 62 , R 63 , and R 64 The naming convention "heteroalkynyl" is used. In certain embodiments, R 57 , R 58 , R 59 , are independently of hydrogen and C 1-3 Selected from alkyl groups.

[0066] "Heteroaryl" refers to a monovalent heteroaromatic radical derived from the removal of one hydrogen atom from one atom of a parent heteroaromatic ring system, either by itself or as part of another substituent. Heteroaryl encompasses at least one heteroaromatic ring fused to a heterocyclic system having at least one other ring, which may be aromatic or nonaromatic. Heteroaryl encompasses a 5- to 7-membered aromatic, monocyclic ring containing one or more heteroatoms selected from N, O, and S, e.g., 1 to 4 or 1 to 3 in certain embodiments, with the remaining ring atoms being carbon; and a bicyclic heterocycloalkyl ring containing one or more heteroatoms selected from N, O, and S, e.g., 1 to 4 or 1 to 3 in certain embodiments, with the remaining ring atoms being carbon, and at least one heteroatom present in the aromatic ring. For example, heteroaryl includes a 5- to 7-membered heteroaromatic ring fused to a 5- to 7-membered cycloalkyl ring. In condensation and bicyclic heteroaryl ring systems where only one of the rings contains one or more heteroatoms, the bonding site may be on the heteroaromatic ring or the cycloalkyl ring. In certain embodiments, if the total number of N, S, and O atoms of the heteroaryl group is greater than 1, the heteroatoms are not adjacent to each other. In certain embodiments, the total number of N, S, and O atoms of the heteroaryl group is 2 or less. In certain embodiments, the total number of N, S, and O atoms of the aromatic heteroring is 1 or less. Heteroaryls do not include or overlap with aryls as defined herein.

[0067] Examples of heteroaryl groups include, but are not limited to, groups derived from acridine, arsindol, carbazole, β-carbolin, chroman, chromene, cinnoline, furan, imidazole, indazole, indole, indoline, indidine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine, phenanthoridine, phenanthroline, phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolidine, quinazoline, quinoline, quinolidine, quinoxaline, tetrazoline, thiadiazole, thiazole, thiophene, triazole, xanthene, etc. In certain embodiments, the heteroaryl group is a 5-membered to 20-membered heteroaryl, and in certain embodiments In some embodiments, the heteroaryl group is a 5- to 10-membered heteroaryl group, and in certain embodiments, it is a 6- to 8-heteroaryl group. In certain embodiments, the heteroaryl group is derived from thiophene, pyrrole, benzothiophene, benzofuran, indole, pyridine, quinoline, imidazole, oxazole, or pyrazine.

[0068] A "heteroarylalkyl" represents an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom is replaced by a heteroaryl group, either by itself or as part of another substituent. Typically, these are terminal or sp 3A heteroarylalkyl group is an atom in which a carbon atom is substituted with a heteroaryl group. When a specific alkyl moiety is intended, the names "heteroarylalkanyl," "heteroarylalkenyl," and "heteroarylalkynyl" are used. In certain embodiments, a heteroarylalkyl group is a 6- to 30-membered heteroarylalkyl group, for example, a heteroarylalkyl group consisting of 1 to 10 alkanyl, alkenyl, or alkynyl moieties and a 5- to 20-membered heteroaryl moiety; and in certain embodiments, a 6- to 20-membered heteroarylalkyl group, for example, a heteroarylalkyl group consisting of 1 to 8 alkanyl, alkenyl, or alkynyl moieties and a 5- to 12-membered heteroaryl moiety.

[0069] "Hypercycloalkyl" refers to a partially saturated or unsaturated cyclic alkyl radical in which one or more carbon atoms (and any associated hydrogen atoms) independently replace the same or different heteroatom group, either by themselves or as part of another substituent. Examples of heteroatoms that replace the carbon atoms include, but are not limited to, N, P, O, S, and Si. When a specific level of saturation is intended, the naming "heterocycloalkanyl" or "heterocycloalkenyl" is used. Examples of heterocycloalkyl groups include, but are not limited to, groups derived from epoxides, azirine, thiirane, imidazolidine, morpholine, piperazine, piperidine, pyrazolidine, pyrrolidine, and quinuclidine.

[0070] "Hypercycloalkylalkyl" refers to a carbon atom, typically terminal or sp, either by itself or as part of another substituent. 3This refers to an acyclic alkyl radical in which one of the hydrogen atoms bonded to a carbon atom is replaced by a heterocycloalkyl group. When a specific alkyl moiety is intended, the nomenclature heterocycloalkylalkanyl, heterocycloalkylalkenyl, or heterocycloalkylalkynyl is used. In certain embodiments, a heterocycloalkylalkyl group is a 6- to 30-membered heterocycloalkylalkyl group, for example, a heterocycloalkylalkyl group consisting of 1 to 10 alkanyl, alkenyl, or alkynyl moieties and a heterocycloalkyl group consisting of 5 to 20 members, and in certain embodiments, a 6- to 20-membered heterocycloalkylalkyl group, for example, a heterocycloalkylalkyl group consisting of 1 to 8 alkanyl, alkenyl, or alkynyl moieties and a heterocycloalkyl group consisting of 5 to 12 members.

[0071] "Leaving group" refers to an atom or group that can be substituted by a nucleophile, and such groups include halogens, such as chloro, bromo, fluoro, and iodine, as well as alkoxycarbonyls (e.g., acetoxy), aryloxycarbonyls, mesyloxy, tosyloxy, trifluoromethanesulfonyloxy, aryloxys (e.g., 2,4-dinitrophenoxy), methoxy, and N,O-dimethylhydroxylamino.

[0072] A "philophilic aromatic ring system" refers to an unsaturated or polycyclic ring system having a conjugated π-electron system. Examples of fused ring systems included in the definition of a "philophilic aromatic ring system" are those in which one or more rings are aromatic and one or more rings are saturated or unsaturated, such as fluorene, indane, and indene. Examples include phenalene. Examples of aromatic ring compounds include, but are not limited to, acetantrylene, acetanaphthylene, acetphenantrylene, anthracene, azulene, benzene, chrysene, coronene, fluorantene, fluorene, hexacene, hexaphene, hexylene, as-indacene, s-indacene, indan, indene, naphthalene, octacene, octafen, octalen, ovalen, penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, phenalene, phenanthrene, picene, pleiaden, pyrene, pyranthrene, rubicene, triphenylene, and trinaphthalene.

[0073] A "parent heteroaromatic ring system" refers to an aromatic ring system in which one or more carbon atoms (and any associated hydrogen atoms) are independently replaced by the same or different heteroatoms. Examples of heteroatoms that replace carbon atoms include, but are not limited to, N, P, O, S, and Si. Specifically included in the definition of a "parent heteroaromatic ring system" are fused ring systems in which one or more rings are aromatic and one or more rings are saturated or unsaturated, such as arsindol, benzodioxane, benzofuran, chroman, chromene, indole, indoline, and xanthene. Examples of hetero-aromatic ring systems include, but are not limited to, arsindol, carbazole, β-carbolin, chroman, chromene, cinnoline, furan, imidazole, indazole, indole, indoline, indoridine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, oxazole, perimidine, phenanthoridine, phenanthroline, phenazine, phthalazine, pteridine, purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolidine, quinazoline, quinoline, quinoridine, quinoxaline, tetrazoline, thiadiazole, thiazole, thiophene, triazole, and xanthene.

[0074] "Patient" refers to an animal, preferably a mammal, and especially preferably a human, including people of all ages and genders.

[0075] "Pharmaceutical composition" refers to at least one compound of the present invention and at least one pharmaceutically acceptable vehicle, which, by use therein, allows at least one compound of the present invention to be administered to a patient, come into contact with a tissue or organ, or come into contact with a cell. "Pharmaceutically acceptable" means that it is approved or expected to be approved by a federal or state regulatory agency, or is listed in the United States Pharmacopeia or other generally accepted pharmacopoeias for use in animals and more preferably in humans.

[0076] "Pharmacologically acceptable salts" refer to salts of a compound that retain the desired pharmacological activity of the parent compound. Examples of such salts include: (1) acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or organic acids, such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4 - Formed with toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-octa-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, t-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, etc.; and (2) salts formed when the acidic proton present in the parent compound is replaced by a metal ion, such as an alkali metal ion, an alkaline earth ion, or an aluminum ion; or ethanolamine, diethanolamine, triethanolamine, N-methylglucamine Coordination compounds of organic bases such as these. In certain embodiments, the pharmaceutically acceptable salt is a hydrochloride salt.

[0077] "Pharmacologically acceptable vehicle" refers to a pharmaceutically acceptable diluent, a pharmaceutically acceptable adjuvant, a pharmaceutically acceptable excipient, a pharmaceutically acceptable carrier, or any combination thereof, which may be used to administer the compound of the present invention to a patient, and which does not destroy the pharmacological activity of the compound of the present invention, and is nontoxic when administered in a dose sufficient to provide the compound in a therapeutically effective amount.

[0078] A "prodrug" is a derivative of a drug molecule that requires conversion to release the active drug in the body. Prodrugs are often, but not necessarily, pharmacologically inactive until converted to the parent drug. The compounds of formulas (I), (III), (VI), (VII), and any of their subspecies or chemical species are creatine prodrugs that are metabolized in the patient's body to release creatine.

[0079] The “precursor” refers to a drug and a group typically linked to the drug’s functional group via a bond(s) that can be cleaved under specific conditions of use. The bond(s) between the drug and the precursor may be cleaved enzymatically or non-enzymatically. Under conditions of use, for example, after administration to a patient, the bond(s) between the drug and the precursor can be cleaved, releasing the parent compound. The cleavage of the precursor may proceed spontaneously, such as via hydrolysis, or it may be catalyzed or induced by another activator, such as an enzyme, light, acid, or by changes in or exposure to physical or environmental parameters, such as temperature or pH. The activator may be inherent in the conditions of use, such as enzymes present in the systemic circulation of the patient receiving the prodrug, or it may be the acidic conditions of the stomach or an externally supplied activator.

[0080] A "protecting group" refers to a group of atoms that, when bonded to a reactive group in a molecule, shield, reduce, or inhibit its reactivity. Examples of protecting groups include Wuts and Greene, "Pro." tective Groups in Organic Synthesis," Jo hn Wiley & Sons, 4th ed. 2006; Harrison et al., "Compendium of Organic Synthetic Methods," Vols.1-11, John Wiley & Sons 1971-2003; Larock "Comprehensive Organic Transformations," John Wiley & Sons, 2n d ed. 2000; and Paquette, "Encyclopedia of Reagents for Organic Synthesis," John Wi This can be seen in ley & Sons, 11th ed. 2003. Examples of amino protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl (Boc), trimethylsilyl (TMS), 2-trimethylsilyl-ethanesulfonyl (SES), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (FMOC), and nitroveratryloxycarbonyl (NVOC). Examples of hydroxy protecting groups include, but are not limited to, those in which the hydroxyl group is either acylated or alkylated, such as benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers, and allyl ethers.

[0081] A "solvate" refers to a molecular complex of a compound with one or more solvent molecules in stoichiometric or nonstoichiometric amounts. Such solvent molecules are those commonly used in the pharmaceutical field and known to be non-invasive to the recipient, such as water and ethanol. These are examples. Molecular complexes of a compound or a part of a compound with a solvent can be stabilized by non-conjugated intramolecular forces, such as electrostatic forces, van der Waals forces, or hydrogen bonds. The term "hydrate" refers to a complex in which one or more solvent molecules are water, and includes monohydrates and hemihydrates.

[0082] "Substantially a single diastereomer" refers to a compound having two or more chiral centers whose diastereomer excess (de) is greater than 90% or at least about 90%. In certain embodiments, the de is, for example, greater than or at least about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%.

[0083] "Substituted" refers to a group in which one or more hydrogen atoms are independently replaced by the same or different substituent(s). Examples of substituents include -M and -R. 70 , -O - ,=O,-OR 70 -SR. 70 , -S - ,=S,-NR 70 R 71 ,=NR 70 , -CF3, -CN, -OCN, -SCN, -NO, -NO2, =N2, -N3, -S(O)2O - -S(O)2OH, -S(O)2R 70 -OS(O2)O - -OS(O)2R 70 ,-P(O)(O - )2, -P(O)(OR 70 )(O - ), -OP(O)(OR 70 )(OR 71 ), -C(O)R 70 ,-C(S)R 70 , -C(O)OR 70 -C(O)NR 70 R 71 -C(O)O-, -C(S)OR 70 , -NR 72 C(O)NR 70 R 71, -NR 72 C(S)NR 70 R 71 , -NR 72 C(NR 73 )NR 70 R 71 , and -C(NR 72 )NR 70 R 71 These are some examples, but are not limited to them, and in the formula, M is independently a halogen; R 70 , R 71 , R 72 , and R 73 Each of these is independently selected from hydrogen, alkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, or R 70 and R 71 These, together with the nitrogen atom to which they are bonded, form a ring selected from a heterocycloalkyl ring. In a particular embodiment, R 70 , R 71 , R 72 , and R 73 These are, independently, hydrogen and C 1-6 Alkyl, C 1-6 Alkoxy, C 3-12 Cycloalkyl, C 3-12 Heterocycloalkyl, C 6-12 Aryl, and C 6-12 Selected from heteroaryls. In certain embodiments, each substituent is independently halogen, -OH, -CN, -CF3, =O, -NO2, C 1-3 Alkoxy, C 1-3 Alkyl, -COOR 80 Selected from, in the formula, R 80 is hydrogen, C 1-3 Alkyl, and (NR 74 ) Selected from 2, in the formula, each R 74 These are, independently, hydrogen or C 1-3 It is alkyl.

[0084] In certain embodiments, substituted aryls and substituted heteroaryls include one or more of the following substituents: F, Cl, Br, C 1-3 Alkyl, substituted alkyl, C1-3 Alkoxy, -S(O)2NR 50 R 51 , -NR 50 R 51 -CF3, -OCF3, -CN, -NR 50 S(O)2R 51 , -NR 50 C(O)R 51 , C 5-10 Aryl substitution C 5-10 Ariel, C 5-10 Heteroaryl, substituted C 5-10 Heteroaryl, -C(O)OR 50 -NO2, -C(O)R 50 -C(O)NR 50 R 51 -OCHF2,C 1-3 Asil, -SR 50 -S(O)2OH, -S(O)2R 50 ,-S(O)R 50 ,-C(S)R 50 ,-C(O)O - , -C(S)OR 50 , -NR 50 C(O)NR 51 R 52 , -NR 50 C(S)NR 51 R 52 , and -C(NR 50 )NR 51 R 52 , C 3-8 Cycloalkyl and substituted C 3-8 Cycloalkyl, in the formula, R 50 , R 51 , and R 52 These are, independently, hydrogen and C 1-4 Selected from alkyl groups.

[0085] In certain embodiments, the substituents are halogen, -NO2, -OH, -COOH, -NH2, -CN, -CF3, -OCF3, C 1-8 Alkyl, substituted C 1-8 Alkyl, C 1-8 Alkoxy and substituted C 1-8 You can choose from alkoxy, and substitute C 1-8 Alkyl and substituted C1-8 Each substituent of the alkoxy can independently be a halogen, -NO2, -OH, -COOH, -NH2, -CN, -CF3, -OCF3 are selected.

[0086] In a particular embodiment, each substituent is independently halogen, -OH, -CN, -CF3, =O, -NO2, C 1-3 Alkoxy, C 1-3 Alkyl, -COOR 80 Selected from, in the formula, R 80 is hydrogen, C 1-3 Alkyl, and (NR 74 ) Selected from 2, in the formula, each R 74 These are, independently, hydrogen or C 1-3 It is alkyl.

[0087] The “therapeutic dose” refers to the amount of a compound sufficient to have an effect on such treatment of a disease, disorder, or symptom when administered to a subject for the treatment of at least one of the clinical symptoms of a disease or disorder. The “therapeutic dose” may vary depending, for example, the compound, the disease, disorder, and / or the symptoms of the disease or disorder, the severity of the symptoms of the disease, disorder, and / or the symptoms of the disease or disorder, the age, weight, and / or health condition of the patient being treated, and the judgment of the prescribing physician. Whatever the case may be, the appropriate dose can be readily determined by a person skilled in the art or can be determined by routine experiments.

[0088] A "therapeutic dose" is the dose that is effective in treating a patient's disease or disorder. The therapeutic dose may vary depending on the compound, the patient, and factors such as the patient's condition and the delivery route. The therapeutic dose can be determined according to commonly used pharmacological procedures known to those skilled in the art.

[0089] "To treat" or "to cure" any disease or disorder means to stop or induce remission of the disease, disorder, or at least one of the clinical symptoms of the disease or disorder, to reduce the risk of developing the disease, disorder, or at least one of the clinical symptoms of the disease or disorder, to slow the progression of the disease, disorder, or at least one of the clinical symptoms of the disease or disorder, or to reduce the risk of progression of the disease, disorder, or at least one of the clinical symptoms of the disease or disorder. "To treat" or "to cure" also means to inhibit the disease or disorder either or both physically (e.g., stabilization of identifiable symptoms) or physiologically (e.g., stabilization of physical parameters), and to inhibit at least one physical parameter, whether identifiable to the patient or not. In certain embodiments, "to treat" or "to cure" means to delay the onset of the disease or disorder or one or more of its symptoms in a patient who is exposed to or predisposed to the disease or disorder, even if the patient does not have or present with the disease or disorder.

[0090] From here, certain embodiments of the compounds, compositions, and methods will be described in detail. The disclosed embodiments are not intended to limit the claims. On the contrary, the claims are intended to encompass all alternative, modified, and equivalent forms.

[0091] Creatine Prodrugs In a particular embodiment, the creatine prodrug is a compound of formula (I), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (I) is as follows: JPEG2026048846000061.jpg71128 During the ceremony: R is either -CH3 or -CD3; R 1 is hydrogen, -OR 2 , -C(O)OR 2 , -C(O)R 2 , TIFF2026048846000062.tif2671 , TIFF2026048846000063.tif2657 , JPEG2026048846000064.jpg2737 ,or JPEG2026048846000065.jpg1752 and; n is an integer between 1 and 2; Each R 2 These are, independently, hydrogen and C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl; Each R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl; R 23 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 5-12 Cycloalkyl, substituted C 5-12 Cycloalkyl, C5-12 Aryl, and C 5-12 Substitutive aryl, -C(O)-OR 22 , or -C(O)-R 22 and; R 22 C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl; and R 48 C 1-12 Alkyl or substituted C 1-12 It is alkyl.

[0092] In certain embodiments of the compound of formula (I), n is an integer 1.

[0093] In certain embodiments of the compound of formula (I), n is an integer 2.

[0094] In certain embodiments of the compound of formula (I), each R 2 and R 22These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-dimethoxyethyl, 1,1-diethoxyethyl, phenyl, 4-methoxyphenyl, benzyl, phenethyl, styryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl.

[0095] In certain embodiments of the compound of formula (I), each R 2 and R 22 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-diethoxyethyl, phenyl, cyclohexyl, or 3-pyridyl.

[0096] In certain embodiments of the compound of formula (I), each R 2 and R 22 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, dodecyl, tert-butyl, phenyl, or cyclohexyl.

[0097] In certain embodiments of the compound of formula (I), each R 2 and R 22 These are independently ethyl, isopropyl, or dodecyl.

[0098] In certain embodiments of the compound of formula (I), R 3 and R 4 Each of them is independently hydrogen.

[0099] In certain embodiments of the compound of formula (I), R 23 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl.

[0100] In certain embodiments of the compound of formula (I), R 23 It is methyl.

[0101] In certain embodiments of the compound of formula (I), each substituent is independently halogen, -NO2, -OH, -NH2, -CN, -CF3, -OCF3, =O, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Alkoxy or substituted C 1-12 Alkoxy, -COOR 10’ And in the formula, R 10’ is hydrogen, C 1-3 Alkyl, or -(NR) 11’ )2, and in the formula, each R 11’ These are, independently, hydrogen or C 1-3 It is alkyl.

[0102] Depending on the embodiment, the compound of formula (I) is a compound of formula (X), formula (XI), formula (XII), formula (XIII), formula (XIV), formula (XV), formula (XVa), or formula (XVb), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof; the compound of formula (X) is the following: JPEG2026048846000066.jpg73128 In the formula, R is either -CH3 or -CD3; The compound of formula (XI) is as follows: JPEG2026048846000067.jpg72128 In the formula, R is -CH3 or -CD3; and R 24 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; The compound of formula (XII) is as follows: JPEG2026048846000068.jpg72128 In the formula, R is -CH3 or -CD3; and R 25 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; The compound of formula (XIII) is as follows: JPEG2026048846000069.jpg73128 In the formula, R is -CH3 or -CD3; and R 26 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; The compound of formula (XIV) is as follows: JPEG2026048846000070.jpg32128 In the formula, R is either -CH3 or -CD3; The compound of formula (XV) is as follows: JPEG2026048846000071.jpg32128 In the formula, R is either -CH3 or -CD3; The compound of formula (XVa) is as follows: JPEG2026048846000072.jpg68125 In the formula, R is either -CH3 or -CD3; R 39 is hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl, and R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl; The compound of formula (XVb) is as follows: JPEG2026048846000073.jpg52128 In the formula, R is either -CH3 or -CD3; R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 alkyl and R 53 C 1-12 Alkyl or substituted C 1-12 It is alkyl.

[0103] In certain embodiments of the compounds of formulas (XI), (XII), and (XIII), each R 24 , R 25 , and R 26 These are independently ethyl, isopropyl, or dodecyl.

[0104] In certain embodiments of the compound of formula (XVa), R 39 These are methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl.

[0105] In certain embodiments of the compound of formula (XVa), R 39 It is methyl.

[0106] In certain embodiments, a compound of formula (XVa) or (XVb) is R 3 and R 4 teeth These are both hydrogen atoms.

[0107] In certain embodiments of the compound of formula (XVb), R 53 These are methyl, ethyl, n-propyl, isopropyl, or tert-butyl.

[0108] In a particular embodiment, the creatine prodrug is a compound of formula (III), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (III) is as follows: JPEG2026048846000074.jpg41128 During the ceremony: W is -CH2OH or -C(O)OR 7 and; R is either -CH3 or -CD3; R 7 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl, substituted C 6-20 Heteroarylalkyl, -C(O)R 5 , -C(O)OR 5 -C(O)(NR 3 R 4 ), -C(R 3 R 4 )-C(O)OR 22 , -C(R 3 R 4 )-(O)C(O)R 22 , -C(R 3 R 4 )-(O)C(O)-OR 22 , TIFF2026048846000075.tif3260 , TIFF2026048846000076.tif3245 ,or JPEG2026048846000077.jpg2323 and; n is an integer between 1 and 2; Each R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 alkyl and R 5 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl; R 23 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 5-12 Cycloalkyl, substituted C 5-12 Cycloalkyl, C 5-12 Aryl, and C 5-12 Substitutive aryl, -C(O)-OR 22 , or -C(O)-R 22 and R 22 C 1-12 Alkyl, substituted C1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl group.

[0109] In certain embodiments of the compound of formula (III), n is an integer 1.

[0110] In certain embodiments of the compound of formula (III), n is an integer 2.

[0111] In certain embodiments of the compound of formula (III), each R 5 , R 7 , and R 22 Independently, C 1-6 Alkyl, substituted C 1-6 Alkyl, C 3-7 Cycloalkyl, substituted C 3-7 Cycloalkyl, C 5-7 Aryl, or substitution C 5-7 It is Ariel.

[0112] In certain embodiments of the compound of formula (III), each R 5 , R 7 , and R 22These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-dimethoxyethyl, 1,1-diethoxyethyl, phenyl, 4-methoxyphenyl, benzyl, phenethyl, styryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl.

[0113] In certain embodiments of the compound of formula (III), each R 5 , R 7 , and R 22 teeth, These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-diethoxyethyl, phenyl, cyclohexyl, or 3-pyridyl.

[0114] In certain embodiments of the compound of formula (III), each R 5 , R 7 , and R 22 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, dodecyl, tert-butyl, phenyl, or cyclohexyl.

[0115] In certain embodiments of the compound of formula (III), each R 5 , R 7 , and R 22 These are independently ethyl, isopropyl, or dodecyl.

[0116] In certain embodiments of the compound of formula (III), each R 3 and R 4 It is, independently, hydrogen.

[0117] In certain embodiments of the compound of formula (III), each R 23These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl.

[0118] In certain embodiments of the compound of formula (III), each R 23 It is methyl.

[0119] In certain embodiments of the compound of formula (III), each substituent is independently halogen, -NO2, -OH, -NH2, -CN, -CF3, -OCF3, =O,C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Alkoxy or substituted C 1-12 Alkoxy, -COOR 10’ And in the formula, R 10’ is hydrogen, C 1-3 Alkyl, or -(NR) 11’ )2, and in the formula, each R 11’ These are, independently, hydrogen or C 1-3 It is alkyl.

[0120] In yet another embodiment, the compound of formula (III) is a compound of formula (XVII), formula (XVIII), or formula (XIX), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof; The compound of formula (XVII) is as follows: JPEG2026048846000078.jpg41128 In the formula, R is either -CH3 or -CD3; R 29 The compounds are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, -cyclohexyl, -CH2-C(O)OR 43 -CH2-(O)C(O)R 43 -CH2-(O)C(O)OR 43 ,or JPEG2026048846000079.jpg2522 and; R 39 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; R 43 is hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; and R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl; The compound of formula (XVIII) is as follows: JPEG2026048846000080.jpg33128 In the formula, R is either -CH3 or -CD3; The compound of formula (XIX) is as follows: JPEG2026048846000081.jpg33128 In the formula, R is either -CH3 or -CD3.

[0121] In certain embodiments of the compound of formula (XVII), each R 29 and R 43 These are independently ethyl, isopropyl, or dodecyl.

[0122] In certain embodiments of the compound of formula (XVII), each R 39 is methyl. In certain embodiments of the compound of formula (XVII), R 3 and R 4 These are hydrogen atoms, respectively.

[0123] In a particular embodiment, the creatine prodrug is a compound of formula (VI), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (VI) is as follows: JPEG2026048846000082.jpg36128 During the ceremony: R is either -CH3 or -CD3; R 10 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl, substituted C 6-20 Heteroarylalkyl, -C(O)R 5 , -C(O)OR 5 -C(O)(NR 3 R 4 ), -C(R 3 R 4 )-C(O)OR 22 , -C(R 3 R 4 )-(O)C(O)R 22 , -C(R 3 R 4 )-(O)C(O)-OR 22 ; JPEG2026048846000083.jpg2323 , TIFF2026048846000084.tif2659 ,or TIFF2026048846000085.tif2644 and; R 11 and R 12 are each independently hydrogen or -OR 13 or; R 11 and R 12 are each -C(O)R 5 wherein R 11 and R 12 cannot both be hydrogen; R 13 is independently hydrogen, C 1-12 alkyl, substituted C 1-12 alkyl, C 1-1 2 heteroalkyl, substituted C 1-12 heteroalkyl, C 3-12 cycloalkyl, substituted C 3-12 cycloalkyl, C 4-20 cycloalkylalkyl, substituted C 4-20 cycloalkylalkyl, C 4-20 heterocycloalkylalkyl, substituted C 4-20 heterocycloalkylalkyl, C 5-12 aryl, substituted C 5-12 aryl, C 5-12 heteroaryl, substituted C 5-12 heteroaryl, C 6-20 arylalkyl, substituted C 6-20 arylalkyl, C 6-20 heteroarylalkyl, substituted C 6-20 heteroarylalkyl -CH(OR 5 ), -C(O)R 5 , -C(O)OR 5 , or -C(O)(NR 3 R 4 ); each R 3 and R 4 is independently hydrogen, C 1-12 alkyl, or substituted C 1-12 alkyl; R 5 is hydrogen, C 1-12 alkyl, substituted C 1-12 alkyl, C 1-12Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl; R 23 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 5-12 Cycloalkyl, substituted C 5-12 Cycloalkyl, C 5-12 Aryl, and C 5-12 Substitutive aryl, -C(O)-OR 22 , or -C(O)-R 22 and; R 22 C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20Arylalkyl, C 6-20 heteroarylalkyl, or substituted C 6-20 heteroarylalkyl; and n is an integer from 1 to 2.

[0124] In certain embodiments of the compound of formula (VI), each R 5 , R 10 , and R 22 is independently C 1-6 alkyl, substituted C 1-6 alkyl, C 3-7 cycloalkyl, substituted C 3-7 cycloalkyl, C 5-7 aryl, or substituted C 5-7 aryl.

[0125] In certain embodiments of the compound of formula (VI), each R 5 , R 10 , and R 22 is independently hydrogen, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-dimethoxyethyl, 1,1-diethoxyethyl, phenyl, p-methoxyphenyl, benzyl, phenethyl, styryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl.

[0126] In certain embodiments of the compound of formula (VI), each R 5 , R 10 , and R 22 is independently hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-diethoxyethyl, phenyl, cyclohexyl, or 3-pyridyl.

[0127] In certain embodiments of the compound of formula (VI), each R 5 , R10 , and R 22 teeth, These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, dodecyl, tert-butyl, phenyl, or cyclohexyl.

[0128] In certain embodiments of the compound of formula (VI), each R 5 , R 10 , and R 22 These are independently ethyl, isopropyl, or dodecyl.

[0129] In certain embodiments of the compound of formula (VI), each R 3 and R 4 It is, independently, hydrogen.

[0130] In certain embodiments of the compound of formula (VI), R 11 and R 12 These are hydroxyls, respectively.

[0131] In certain embodiments of the compound of formula (VI), R 11 or R 12 One of them is hydrogen, and the other is hydroxyl.

[0132] In certain embodiments of the compound of formula (VI), each R 23 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl.

[0133] In certain embodiments of the compound of formula (VI), each R 23 It is methyl.

[0134] In certain embodiments of the compound of formula (VI), each substituent is independently halogen, -NO2, -OH, -NH2, -CN, -CF3, -OCF3, =O,C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Alkoxy or substituted C 1-12Alkoxy, -COOR 10’ And in the formula, R 10’ is hydrogen, C 1-3 Alkyl, or -(NR) 11’ )2, and in the formula, each R 11’ These are, independently, hydrogen or C 1-3 It is alkyl.

[0135] In certain embodiments of the compound of formula (VI), n is an integer 1.

[0136] In yet another embodiment, the compound of formula (VI) is a compound of formula (XXII), formula (XXIII), formula (XXIV), formula (XXV), formula (XXVI), formula (XXVII), or formula (XXVIII), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof; The compound of formula (XXII) is as follows: JPEG2026048846000086.jpg36128 The compound of formula (XXIII) is as follows: JPEG2026048846000087.jpg36128 The compound of formula (XXIV) is as follows: JPEG2026048846000088.jpg36128 The compound of formula (XXV) is as follows: JPEG2026048846000089.jpg32128 The compound of formula (XXVI) is as follows: JPEG2026048846000090.jpg37128 The compound of formula (XXVII) is as follows: JPEG2026048846000091.jpg36128 The compound of formula (XXVIII) is as follows: JPEG2026048846000092.jpg41128 In the formula, R is either -CH3 or -CD3; R a These are hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl; R 32 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, and cyclohexyl; -CH2-C(O)OR 43 -CH2-(O)C(O)R 43 -CH2-(O)C(O)OR 43 ,or JPEG2026048846000093.jpg2522 and; R 39 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; Each R 33 R is independently hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; 43 is hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl; and R 3 and R 4 These are, independently, hydrogen and C 1-12 Alkyl or substituted C 1-12 It is alkyl.

[0137] In a particular embodiment, each R 32 and R 33 These are independently ethyl, isopropyl, or dodecyl.

[0138] In a particular embodiment, R 39It is methyl.

[0139] In a particular embodiment, R 3 and R 4 These are hydrogen atoms, respectively.

[0140] In a particular embodiment, the creatine prodrug is a compound of formula (VII), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (VII) is as follows: JPEG2026048846000094.jpg38128 During the ceremony: R is either -CH3 or -CD3; Each R 14 These are, independently, hydrogen and C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl, substituted C 6-20 Heteroarylalkyl-CH(OR 5 ), -C(O)R 5 , -C(O)OR 5 , or -C(O)(NR 3 R 4 ) and; Each R 3 and R 4 These are, independently, hydrogen and C1-12 Alkyl or substituted C 1-12 alkyl and R 5 is hydrogen, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Heteroalkyl, substituted C 1-12 Heteroalkyl, C 3-12 Cycloalkyl, substituted C 3-12 Cycloalkyl, C 4-20 Cycloalkylalkyl, substituted C 4-20 Cycloalkylalkyl, C 4-20 Heterocycloalkylalkyl, substituted C 4-20 Heterocycloalkylalkyl, C 5-12 Aryl substitution C 5-12 Ariel, C 5-12 Heteroaryl, substituted C 5-12 Heteroaryl, C 6-20 Arylalkyl, substituted C 6-20 Arylalkyl, C 6-20 Heteroarylalkyl or substituted C 6-20 It is a heteroarylalkyl group.

[0141] In certain embodiments of the compound of formula (VII), R 5 C 1-6 Alkyl, substituted C 1-6 Alkyl, C 3-7 Cycloalkyl, substituted C 3-7 Cycloalkyl, C 5-7 Aryl, or substitution C 5-7 It is Ariel.

[0142] In certain embodiments of the compound of formula (VII), R 5These are hydrogen, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-dimethoxyethyl, 1,1-diethoxyethyl, phenyl, 4-methoxyphenyl, benzyl, phenethyl, styryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl.

[0143] In certain embodiments of the compound of formula (VII), R 5 These are hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-diethoxyethyl, phenyl, cyclohexyl, or 3-pyridyl.

[0144] In certain embodiments of the compound of formula (VII), R 5 These are hydrogen, methyl, and ethyl These are n-propyl, isopropyl, dodecyl, tert-butyl, phenyl, or cyclohexyl.

[0145] In certain embodiments of the compound of formula (VII), R 5 It is ethyl, isopropyl, or dodecyl.

[0146] In certain embodiments of the compound of formula (VII), each R 14 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl.

[0147] In certain embodiments of the compound of formula (VII), one R 14 is methyl, and the other R 14 It is hydrogen.

[0148] In certain embodiments of the compound of formula (VII), each R3 and R 4 It is, independently, hydrogen.

[0149] In certain embodiments of the compound of formula (VII), each substituent is independently halogen, -NO2, -OH, -NH2, -CN, -CF3, -OCF3, =O,C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Alkoxy or substituted C 1-12 Alkoxy, -COOR 10’ And in the formula, R 10’ is hydrogen, C 1-3 Alkyl, or -(NR) 11’ )2, and in the formula, each R 11’ These are, independently, hydrogen or C 1-3 It is alkyl.

[0150] In yet another embodiment, the compound of formula (VII) is the compound of formula (XXIX), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof; The compound of formula (XXIX) is as follows: JPEG2026048846000095.jpg36128 In the formula, R is -CH3 or -CD3; and Each R 34 These are independently hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl.

[0151] In certain embodiments of the compound of formula (XXIX), one R 34 is methyl, and the other R 34 It is hydrogen.

[0152] Synthesis of creatine prodrugs Those skilled in the art will see that the creatine prodrug compounds of formulas (I), (III), (VI), and (VII), and any variants or chemical species thereof, or their pharmaceutically acceptable salts, solvates, tautomers, or stereoisomers, can be prepared by common synthetic methods available in the art (e.g., Wuts and Greene, "P "rotective Groups in Organic Synthesis," John Wiley & Sons, 4th ed. 2006; n et al., “Compendium of Organic Synthet ic Methods," Vols. 1-11, John Wiley & So ns 1971-2003; Larock "Comprehensive Orga" nic Transformations," John Wiley & Sons, 2nd ed. 2000; and Paquette, "Encyclopedia of Reagents for Organic Synthesis," John Wiley & Sons, 11th ed. 2003). Starting materials useful for preparing compounds and their intermediates are commercially available, or can be prepared by well-known synthetic methods.

[0153] Pharmaceutical composition The pharmaceutical composition of the present invention may contain the compound of the present invention and a pharmaceutically acceptable vehicle. The pharmaceutical composition may contain the compound of the present invention in a therapeutically effective amount and a pharmaceutically acceptable vehicle. In certain embodiments, the pharmaceutical composition may contain more than one of the compounds of the present invention. Examples of pharmaceutically acceptable vehicles include diluents, adjuvants, excipients, and carriers.

[0154] Pharmaceutical compositions can be manufactured using standard procedures (for example, "Remingto n's The Science and Practice of Pharmacy," 21st edition, Lippincott, Williams & (See Wilcox, 2005). Pharmaceutical compositions can be prepared by conventional mixing, dissolution, granulation, dragée formation, levigating, emulsification, encapsulation, encapsulation, or lyophilization processes. Pharmaceutical compositions can be formulated conventionally with one or more physiologically acceptable carriers, diluents, excipients, or auxiliaries, which facilitate the formulation of the compounds disclosed herein into pharmaceutically usable formulations. Appropriate formulation may depend in part on the route of administration.

[0155] The pharmaceutical compositions of the present invention can provide therapeutic plasma concentrations of creatine when administered to a patient. The precursor portion of the creatine prodrug can be chemically and / or enzymatically cleaved in vivo to release creatine. One or more enzymes present in the mammalian intestinal lumen, intestinal tissue, blood, liver, brain, or any other suitable tissue can enzymatically cleave the precursor portion of the administered prodrug. For example, the precursor portion may be cleaved after absorption by the gastrointestinal tract (e.g., in the mammalian intestinal tissue, blood, liver, or other suitable tissue). In certain embodiments, creatine is protected from pre-systemic circulation metabolism by remaining bound to the precursor portion while crossing the intestinal mucosal barrier. In certain embodiments, the creatine prodrug is essentially not metabolized in the intestinal cell to release the corresponding creatine, but is metabolized in the systemic circulation to become the parent compound. Cleavage of the precursor portion of the creatine prodrug after absorption by the gastrointestinal tract may allow the prodrug to be absorbed into the systemic circulation by either active transport, passive diffusion, or a combination of both active and passive processes.

[0156] Creatine prodrugs can remain intact until they have crossed biological barriers, such as the blood-brain barrier. In certain embodiments, the prodrugs of the present invention can be partially cleaved, for example, one or more of the precursor portions can be cleaved before crossing biological barriers or after being taken up by cells, tissues, or organs.

[0157] Creatine prodrugs can remain intact in systemic circulation and can be absorbed by organ cells either by passive or active transport mechanisms. In certain embodiments, creatine prodrugs become lipophilic and are passively transported across cell membranes. It can be moved. After cellular uptake, the prodrug can be chemically and / or enzymatically cleaved to release the corresponding creatine into the cell's cytoplasm, resulting in an increase in the intracellular concentration of creatine. In certain embodiments, the prodrug can permeate intracellular membranes such as the mitochondrial membrane, thereby facilitating the delivery of the prodrug to intracellular organelles such as mitochondria, and subsequent cleavage of the precursor portion or multiple precursor portions, thereby facilitating the delivery of creatine.

[0158] In certain embodiments, the pharmaceutical composition may include an adjuvant that enhances the absorption of the compounds of the present invention through the gastrointestinal epithelium. Such enhancers can, for example, open tight junctions in the gastrointestinal tract or modify the effects of cellular components such as p-glycoproteins. Suitable enhancers include alkali metal salts of salicylic acid, e.g., sodium salicylate, and alkali metal salts of caprylic or capric acid, e.g., sodium caprylate or sodium caprate. As enhancers, bile salts such as sodium deoxycholate can be cited. Various p-glycoprotein modifiers are described in U.S. Patent No. 5,112,817 and U.S. Patent No. 5,643,909. Various absorption-enhancing compounds and materials are described in U.S. Patent No. 5,824,638 and U.S. Patent Application No. 2006 / 0046962. Other adjuvants that improve cell membrane permeability include resorcinol, surfactants, polyethylene glycol, and bile acids.

[0159] In certain embodiments, the pharmaceutical composition may include an adjuvant that reduces the enzymatic degradation of the compound of the present invention. Microencapsulation using proteinoid microparticles, liposomes, or polysaccharides may also be effective in reducing the enzymatic degradation of the administered compound.

[0160] A pharmaceutical composition may also include one or more pharmaceutically acceptable vehicles, such as excipients, adjuvants, carriers, diluents, binders, lubricants, disintegrants, colorants, stabilizers, surfactants, fillers, buffers, thickeners, emulsifiers, and wetting agents. The vehicle may be selected to modify the porosity and permeability of the pharmaceutical composition, to modify its hydration and disintegration properties, to control hydration, to improve manufacturability, and so on.

[0161] In certain embodiments, the pharmaceutical composition is formulated for oral administration. The pharmaceutical composition formulated for oral administration can result in uptake of the compound of the present invention through the gastrointestinal tract, or in one or more specific regions of the gastrointestinal tract. In certain embodiments, the pharmaceutical composition can be formulated to facilitate uptake of the compound of the present invention from the upper gastrointestinal tract, and, in certain embodiments, from the small intestine. Such compositions can be manufactured in ways known in the pharmaceutical field and may further contain, in addition to the compound of the present invention, one or more pharmaceutically acceptable vehicles, permeability enhancers, and / or secondary therapeutic agents.

[0162] In certain embodiments, the pharmaceutical composition may further include substances that improve, modify, and / or control release, bioavailability, therapeutic efficacy, therapeutic potency, stability, etc. For example, to improve therapeutic efficacy, the compounds of the present invention may be administered co-administered with one or more activators that increase the absorption or diffusion of the drug from the gastrointestinal tract or inhibit the breakdown of the drug in systemic circulation. In certain embodiments, the compounds of the present invention may be administered co-administered with activators having pharmacological effects that improve the therapeutic efficacy of the compounds of the present invention.

[0163] In certain embodiments, the pharmaceutical composition further includes release, bioavailability, and therapeutic efficacy. The compounds may include substances that improve, modify, and / or control therapeutic efficacy, stability, etc. For example, to improve therapeutic efficacy, the compounds of the present invention may be administered co-administered with one or more activators that increase the absorption or diffusion of the compounds of the present invention from the gastrointestinal tract or inhibit the breakdown of the drug in systemic circulation. In certain embodiments, the compounds of the present invention may be administered co-administered with activators that have pharmacological effects that improve the therapeutic efficacy of the compounds of the present invention.

[0164] Pharmaceutical compositions can take the form of liquids, suspensions, emulsions, tablets, pills, pellets, capsules, liquid-containing capsules, powders, sustained-release formulations, suppositories, emulsions, aerosols, sprays, suspensions, or any other form suitable for use. Pharmaceutical compositions for oral delivery can take the form of, for example, tablets, lozenges, aqueous or oily suspensions, granules, powders, emulsions, capsules, syrups, or elixirs. Compositions administered orally may contain one or more optional agonists, such as sweeteners, such as fructose, aspartame, or saccharin; flavoring agents, such as peppermint, wintergreen oil, or cherry coloring agents; and preservatives, in order to provide a pharmaceutically palatable formulation. Furthermore, if in the form of tablets or pills, compositions may be coated to slow down breakdown and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period. Oral compositions may include standard vehicles such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, and magnesium carbonate. Such vehicles may be of pharmaceutical grade. For oral liquid formulations, such as suspensions, elixirs, and liquids, suitable carriers, excipients, or diluents include water, physiological saline, alkylene glycol (e.g., propylene glycol), polyalkylene glycol (e.g., polyethylene glycol), oil, alcohol, and weakly acidic buffers with a pH of 4 to 6 (e.g., acetates, citrates, ascorbic acid salts, etc., at approximately 5 mM to 50 mM). Flavorings, preservatives, colorants, bile salts, and acylcarnitines may also be added.

[0165] If the compound of the present invention is acidic, it may be included in any of the above formulations as a free acid, a pharmaceutically acceptable salt, a solvate, a hydrate, or a hydrate. A pharmaceutically acceptable salt substantially retains the activity of the free acid, can be prepared by reaction with a base, and tends to be more soluble in aqueous solvents and other protic solvents than the corresponding free acid form. In some embodiments, a sodium salt of the compound of the present invention is used in the above formulations.

[0166] The pharmaceutical compositions of the present invention can be formulated for parenteral administration, such as by injection, e.g., intravenous, intraarterial, intramuscular, subcutaneous (as a depot formulation), intrapericardial, coronary artery injection, or use as a solution for delivery to tissues or organs, e.g., use in cardiopulmonary bypass devices or for immersing transplanted tissues or organs. The injectable compositions can be pharmaceutical compositions for any route of injectable administration, such as intravenous, intraarterial, intracoronal, pericardial, perivascular, intramuscular, subcutaneous, intradermal, intraperitoneal, and intraarticular. In certain embodiments, the injectable pharmaceutical composition may be a composition pharmaceutically suitable for direct administration to the heart, pericardium, or coronary artery.

[0167] A pharmaceutical composition of the present invention suitable for parenteral administration may contain one or more compounds of the present invention in combination with one or more pharmaceutically acceptable sterile isotonic aqueous, water-miscible, or non-aqueous vehicles. The pharmaceutical composition for parenteral use may contain substances that increase and maintain drug solubility, such as complexing agents and surfactants; compounds that bring the solution closer to isotonic or physiological pH, such as sodium chloride, glucose, and glycerin; substances that improve the chemical stability of the solution, such as antioxidants, inert gases, chelating agents, and buffers; and chemical and physical stability The parenteral formulation may include substances that improve performance, substances that minimize self-aggregation or interface-induced aggregation, substances that minimize interaction with protein interfaces, preservatives including antimicrobial agents, suspending agents, emulsifiers, and any combination of the above. Parenteral formulations can be formulated as liquids, suspensions, emulsions, liposomes, microparticles, nanosystems, and powders for reconstitution as liquids. Parenteral formulations may include "Remington, The Science and Practice of Pharmacy," 21st edition, Lippincott, Williams & Wilkins, This is described in Chapters 41-42, pages 802-849, 2005.

[0168] In certain embodiments, the pharmaceutical composition can be a bathing formulation for transplanted tissue or organs before, during, or after transfer to the intended recipient. Such a composition can be used before or during the preparation of tissue or organs for transplantation. In certain embodiments, the pharmaceutical composition may be a cardiac arrest solution administered during cardiac surgery. In certain embodiments, the pharmaceutical composition can be used, for example, in combination with a cardiopulmonary bypass device to deliver the pharmaceutical composition to the heart. Such a pharmaceutical composition can be used during the induction, maintenance, or reperfusion phases of cardiac surgery (e.g., Chang et al., Masui 2003, 52(4), 356-62; Ibrahim et al., Eur. J. Cardiothorac Surg 1999, 15(1), 75-83; von Oppell et al., J Thorac Cardiovasc Surg. 1991, 102(3), 405-12; and Ji et al., J. Extra Corpor Technol See 2002, 34(2), 107-10). In certain embodiments, the pharmaceutical composition may be delivered via mechanical devices such as pumps or perfusion devices (see, for example, Hou and March, J Invasive Cardiol 2003, 15(1), 13-7; Maisch et al., Am. J Cardiol 2001, 88(11), 1323-6; and Macris and Igo, Clin Cardiol 1999, 22(1, Suppl 1), 136-9).

[0169] For long-term delivery, the pharmaceutical composition can be provided as a depot formulation for administration by implantation, for example, subcutaneous, intradermal, or intramuscular injection. That is, in certain embodiments, the pharmaceutical composition can be formulated, for example, as an emulsion contained in a pharmaceutically acceptable oil or ion exchange resin, together with a suitable polymer or hydrophobic material, or in the form of a poorly soluble derivative, for example, a poorly soluble salt of the compound of the present invention.

[0170] The pharmaceutical compositions of the present invention can be formulated using procedures known in the art to provide immediate, sustained, or delayed release of compounds of formula (I) and / or formula (II) after administration to a patient (e.g., Allen et al., "Ansel's Phar"). Maceutical Dosage Forms and Drug Delivery Systems," 8th ed., Lippincott, William (See s & Wilkins, August 2004).

[0171] Dosage form The pharmaceutical compositions of the present invention can be formulated in unit dosage forms. A unit dosage form represents a physically isolated unit appropriate as a unit dose for a patient receiving treatment, and each unit contains a predetermined amount of the compound of the present invention calculated to produce the desired therapeutic effect. A unit dosage form may be for a once-daily dose or for multiple doses per day, for example, 2 to 4 times per day. When multiple doses per day are used, the unit dosage forms may consist of identical or different individual doses. One or more dosage forms may form a single dose. This may be possible, and this single dose may be administered to the patient at a specific point in time or over a period of time.

[0172] The pharmaceutical compositions of the present invention can be used in dosage forms that provide immediate and / or controlled release of the compounds of the present invention. The appropriate dosage form may depend on the disease, disorder, or symptom being treated and the method of administration. For example, for the treatment of acute ischemic symptoms such as heart failure or stroke, the use of an immediate-release pharmaceutical composition or dosage form administered parenterally may be appropriate. For the treatment of chronic neurodegenerative diseases, an orally administered controlled-release pharmaceutical composition or dosage form may be appropriate.

[0173] In certain embodiments, the dosage form can be adapted for administration to a patient twice or less per day, and in certain embodiments, only once per day. The medication can be provided alone or in combination with other drugs and can be continued for as long as necessary for the effective treatment of the disease, disorder, or symptoms.

[0174] Pharmaceutical compositions containing the compounds of the present invention can be formulated for parenteral administration, oral administration, or immediate release via any other suitable route of administration.

[0175] Controlled drug delivery systems can be designed to deliver drugs in a way that maintains drug levels within the therapeutic range and effective and safe blood levels throughout the delivery period, as long as the delivery system continues to deliver the drug at a specific rate. Controlled drug delivery can result in substantially constant drug blood levels compared to the fluctuations observed with immediate-release formulations. For some drugs, maintaining constant blood flow and tissue concentrations throughout the entire course of treatment is the most desirable mode of therapy. Immediate release of such drugs can result in peak blood levels exceeding the levels necessary to induce the desired response, which can waste the drug and cause or exacerbate toxic side effects. Controlled drug delivery can result in optimal treatment, reducing the frequency of dosing and potentially lowering the severity of side effects. Examples of controlled-release formulations include solubility-controlled systems, diffusion-controlled systems, ion-exchange resins, osmotic pressure-controlled systems, erosion-type matrix systems, pH-independent formulations, and gastric retention systems.

[0176] In certain embodiments, the oral dosage form of the present invention can be a controlled-release dosage form. Controlled delivery techniques can improve the absorption of a drug in one or more specific areas of the gastrointestinal tract. The appropriate oral dosage form for a particular pharmaceutical composition of the present invention may depend, at least in part, on the gastrointestinal absorption properties of the compound of the present invention, the stability of the compound of the present invention in the gastrointestinal tract, the pharmacokinetics of the compound of the present invention, and the desired therapeutic properties. An appropriate controlled-release oral dosage form can be selected for a particular compound of the present invention. For example, a gastric retention oral dosage form may be appropriate for a compound absorbed primarily from the upper gastrointestinal tract, and a sustained-release oral dosage form may be appropriate for a compound absorbed primarily from the lower gastrointestinal tract.

[0177] Certain compounds are primarily absorbed in the small intestine. Generally, compounds travel the length of the small intestine over approximately 3 to 5 hours. For compounds that are not readily absorbed or readily dissolved in the small intestine, the time window for active ingredient absorption in the small intestine may be too short to provide the desired therapeutic effect. Gastric retention formulations, i.e., formulations designed to remain in the stomach for an extended period, can increase the bioavailability of drugs that are most readily absorbed in the upper gastrointestinal tract. Conventional formulations have a gastric residence time of 1 to 3 hours. After passing through the stomach, the bioavailability window until the formulation reaches the colon is approximately 3 to 5 hours. However, when a formulation remains in the stomach, the drug can be released before it reaches the small intestine, entering the intestines as a solution in a state where it can be more readily absorbed. Another use of gastric retention formulations is to improve the bioavailability of drugs that are unstable to the basic conditions of the intestines (e.g., H See Wang et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1998, 15, 243-284. Several gastric retention formulations have been developed to improve drug absorption from the upper gastrointestinal tract. Examples include hydrogels (see, e.g., U.S. Patent Application No. 2003 / 0008007), floating matrices (see, e.g., U.S. Patent Application No. 2006 / 0013876), polymer sheets (see, e.g., U.S. Patent Application No. 2005 / 0249798), microcell foams (see, e.g., U.S. Patent Application No. 2005 / 0202090), and swelling formulations (see, e.g., U.S. Patent Application No. 200 Examples include U.S. Patent No. 5 / 0019409; U.S. Patent No. 6,797,283; U.S. Patent Application No. 2006 / 0045865; U.S. Patent Application No. 2004 / 0219186; U.S. Patent No. 6,723,340; U.S. Patent No. 6,476,006; U.S. Patent No. 6,120,803; U.S. Patent No. 6,548,083; U.S. Patent No. 6,635,280; and U.S. Patent No. 5,780,057. Bioadhesive polymers can also provide vehicles for controlled drug delivery to multiple mucosal surfaces in addition to the gastric mucosa (see, for example, U.S. Patent No. 6,235,313; U.S. Patent No. 6,207,197; U.S. Patent Application No. 2006 / 0045865, and U.S. Patent Application No. 2005 / 0064027). Ion exchange resins have been shown to retain in the stomach for extended periods, which may be due to adhesion.

[0178] In a swelling and expansion system, dosage forms that swell and change density in relation to the surrounding stomach contents can be retained in the stomach for a longer period than conventional dosage forms. The dosage form can absorb water and swell to form a gelatinous outer surface, allowing it to float on the surface of the stomach contents while maintaining its integrity until drug release. If hydration and swelling alone are insufficient, fatty materials can be added to prevent wetting and facilitate buoyancy. Gas-releasing materials may also be incorporated to reduce the density of the gastric retention dosage form. Swelling can also significantly increase the size of the dosage form, thereby preventing the release of undisintegrated swollen solid dosage forms into the small intestine through the pylorus. Swellable dosage forms can be formed by encapsulating a drug-containing nucleus and a swelling agent, or by combining the drug, swelling agent, and one or more erosionable polymers.

[0179] The gastric retention dosage form may also be a folded thin sheet containing the drug and a water-insoluble diffusible polymer, which expands in the stomach to its original dimensions and shape, and the original dimensions and shape are large enough to prevent or inhibit the expanded dosage form from passing through the pyloric sphincter.

[0180] Floating and buoyant gastric retention dosage forms can be designed to retain the dosage form in the stomach for longer periods, for example, 9-12 hours, by encapsulating gas within a sealed nucleus that allows the nucleus to float on the stomach contents. Due to the buoyancy effect, such systems can provide a protective layer to prevent reflux of stomach contents into the esophageal region and can also be used in controlled release devices. A suspension system may contain, for example, a hollow nucleus containing a drug coated with a protective film. The air trapped in the nucleus keeps the dosage form afloat on the stomach contents until the dissolved components are released and the system disintegrates. In other suspension systems, the nucleus contains a drug and a chemical that can produce gas when activated. For example, a coated nucleus containing carbonates and / or bicarbonates can react with hydrochloric acid in the stomach or with organic acids incorporated into the system to produce carbon dioxide. The gas produced by the reaction is retained and keeps the dosage form afloat. The expanded dosage form gradually disintegrates and disappears from the stomach as the generated gas slowly permeates the protective coating.

[0181] Bioadhesive polymers can also provide vehicles for controlled drug delivery to multiple mucosal surfaces in addition to the gastric mucosa (see, for example, U.S. Patent No. 6,235,313 and U.S. Patent No. 6,207,197). The bioadhesive system contains the drug and other excipients within the bioadhesive polymer. It can be designed by incorporating the following. Upon ingestion, the polymer hydrates and adheres to the mucosa of the gastrointestinal tract. The bioadhesive polymer can be selected to adhere to one or more desired locations in the gastrointestinal tract. The bioadhesive polymer can be selected to be optimally delivered to target areas in the gastrointestinal tract, including the stomach and small intestine. The mechanism of adhesion is thought to be through the formation of electrostatic and hydrogen bonds at the polymer mucosal boundary. U.S. Patent Applications 2006 / 0045865 and 2005 / 0064027 disclose bioadhesive delivery systems useful for drug delivery to both the upper and lower gastrointestinal tract.

[0182] Ion exchange resins have been shown to remain in the stomach for extended periods, which may be due to adhesion.

[0183] Gastric retention oral dosage forms can be appropriately used for the delivery of drugs that are primarily absorbed through the upper gastrointestinal tract. For example, certain compounds of the present invention may exhibit limited colonic absorption and may be primarily absorbed through the upper gastrointestinal tract. That is, dosage forms that release the compounds of the present invention in the upper gastrointestinal tract and / or delayed transfer of dosage forms through the upper gastrointestinal tract tend to improve the oral bioavailability of the compounds of the present invention. Other forms of creatine prodrugs disclosed herein can be appropriately used in gastric retention dosage forms.

[0184] Polymer matrices have also been used to achieve controlled release of drugs over extended periods. Such sustained or controlled release can be achieved by limiting the rate at which ambient gastric juice can diffuse into the matrix to reach the drug, dissolve the drug, and re-diffuse with the dissolved drug, or by using a matrix that is slowly eroded, continuously exposing fresh drug to the ambient fluid. Disclosures of polymer matrices that function in these ways can be found, for example, in Skinner, U.S. Patents 6,210,710 and 6,217,903; U.S. Patents 5,451,409; U.S. Patents 5,945,125; PCT International Publication WO96 / 26718; U.S. Patents 4,915,952; U.S. Patents 5,328,942; U.S. Patents 5,783,212; U.S. Patents 6,120,803; and U.S. Patents 6,090,411.

[0185] Other drug delivery devices that remain in the stomach for extended periods include, for example, particle-containing hydrogel reservoirs (U.S. Patent No. 4,871,548); swellable hydroxypropyl methylcellulose polymers (U.S. Patent No. 4,871,548); plate-shaped bio-erosive polymers (U.S. Patent No. 4,767,627); multiple compressible retaining arms (U.S. Patent No. 5,443,843); hydrophilic water-swellable crosslinked polymer particles (U.S. Patent No. 5,007,790); and albumin-crosslinked polyvinylpyrrolidone hydrogels (Park et al., J. Controlled Release 1992, 19, 131-134).

[0186] In certain embodiments, the pharmaceutical compositions of the present invention can be implemented in several different dosage forms that can be adapted to provide sustained-release of the compound of the present invention when administered orally. Sustained-release oral dosage forms can be used to release a drug over a long period of time and are useful when it is desirable to deliver the drug or formulation to the lower gastrointestinal tract. Examples of sustained-release oral dosage forms include diffusion-controlled systems, such as reservoir devices or matrix devices, dissolution-controlled systems, osmotic systems, and erosion-controlled systems. Sustained-release oral dosage forms and methods for producing them are well known in the art (e.g., "Remington's Pharmaceuticals"). utical Sciences," Lippincott, Williams & Wilkins, 21st edition, 2005, Chapters 46 and 47; Langer, Science 1990, 249, 1527-1533; and Rosoff, "Controlled Release of See "Drugs," 1989, Chapter 2.

[0187] Sustained-release oral dosage forms include any oral dosage form that maintains the therapeutic concentration of a drug for a long period of time in biological fluids, such as plasma, blood, or cerebrospinal fluid, or in tissues or organs. Examples of sustained-release oral dosage forms include diffusion-controlled systems, such as reservoir devices or matrix devices, dissolution-controlled systems, osmotic pressure systems, and erosion-controlled systems. Sustained-release oral dosage forms and methods for producing them are well known in the art (e.g., "Remington's: T"). he Science and Practice of Pharmacy," Li ppincott, Williams & Wilkins, 21st edition, 2005, Chapters 46 and 47; Langer, Science 1990, 249, 1527-1533; and Rosoff, "Co ntrolled Release of Drugs," 1989, Chapter (See r 2).

[0188] In diffusion-controlled systems, a water-insoluble polymer controls the fluid flow and the subsequent release of the dissolved drug from the dosage form. Both the diffusion and dissolution processes are involved in the release of the drug from the dosage form. In reservoir devices, drug-containing nuclei are coated with a polymer, while in matrix systems, the drug is dispersed throughout the matrix. Cellulose polymers, such as ethylcellulose or cellulose acetate, can be used in reservoir devices. Examples of materials useful for matrix systems include methacrylic acid compounds, acrylic acid compounds, polyethylene, acrylic acid copolymers, polyvinyl chlorides, high molecular weight polyvinyl alcohols, cellulose derivatives, and fatty compounds, such as fatty acids, glycerides, and carnauba wax.

[0189] In dissolution-controlled systems, the dissolution rate of a drug is controlled by a slow-dissolving polymer or by microencapsulation. Once the coating dissolves, the drug becomes soluble. The drug release rate can be controlled by changing the thickness and / or composition of one or more coatings. Some dissolution-controlled systems may include an immediate-release component in part of the total dose. Examples of dissolution-controlled systems include encapsulation / reservoir dissolution systems and matrix dissolution systems. Encapsulation dissolution systems can be prepared by coating drug particles or granules with a slow-dissolving polymer of varying thicknesses or by microencapsulation. Examples of coating materials useful in dissolution-controlled systems include gelatin, carnauba wax, shellac, cellulose phthalate acetate, and cellulose butyrate acetate. Matrix dissolution devices can be prepared, for example, by compressing a drug with a slow-dissolving polymer carrier into a tablet form.

[0190] The drug release rate from an osmotic pump system is determined by the inflow of fluid across a semipermeable membrane into a reservoir, which contains an osmotic agent. The drug is either mixed with the agent or located in the reservoir. The dosage form contains one or more small orifices from which the dissolved drug is pumped at a rate determined by the osmotic water inflow rate. As the osmotic pressure within the dosage form increases, the drug is released through the orifice(s). The release rate is constant and can be controlled within strict limits, thus resulting in relatively constant plasma and / or blood concentrations of the drug. The osmotic pump system can provide constant drug release independently of the gastrointestinal environment. The drug release rate can be modified by altering the osmotic agent and the dimensions of one or more orifices.

[0191] Drug release from erosion-controlled systems is determined by the erosion rate of the carrier matrix. The drug is dispersed throughout the polymer, and the drug release rate depends on the erosion rate of the polymer. The drug-containing polymer can be degraded from the active ingredient and / or from the surface of the dosage form.

[0192] Sustained-release oral dosage forms are any form suitable for oral administration, such as tablets, pills, or granules. These are some examples. Granules can be filled into capsules, compressed into tablets, or included in liquid suspensions. Sustained-release oral dosage forms may further include an outer coating to provide, for example, acid protection, ease of swallowing, flavor, and distinctiveness.

[0193] In certain embodiments, a sustained-release oral dosage form may contain a therapeutically effective amount of the compound of the present invention and a pharmaceutically acceptable vehicle. In certain embodiments, a sustained-release oral dosage form may contain a less than therapeutically effective amount of the compound of the present invention and a pharmaceutically acceptable vehicle. Multiple sustained-release oral dosage forms, each containing a less than therapeutically effective amount of the compound of the present invention, can be administered at once or over a period of time to provide a therapeutically effective dose or regimen for treating diseases in patients associated with impaired energy metabolism, such as ischemia, oxidative stress, neurodegenerative diseases including amyotrophic lateral sclerosis (ALS), Huntington's disease, Parkinson's disease, or Alzheimer's disease, ischemia-reperfusion injury, cardiovascular disease, multiple sclerosis (MS), psychiatric disorders, genetic diseases affecting the creatine kinase system, or muscle fatigue.

[0194] The sustained-release oral dosage form of the present invention can release the compound of the present invention from the dosage form to facilitate the ability of the compound of the present invention to be absorbed from a suitable region of the gastrointestinal tract, for example, in the small intestine or the colon. In certain embodiments, the sustained-release oral dosage form can release the compound of the present invention from the dosage form over a period of at least about 4 hours, at least about 8 hours, at least about 12 hours, at least about 16 hours, at least about 20 hours, and in certain embodiments, at least about 24 hours. In certain embodiments, the sustained-release oral dosage form can release the compound of the present invention from the dosage form in a delivery pattern of about 0 wt% to about 20 wt% over about 0 to about 4 hours, about 20 wt% to about 50 wt% over about 0 to about 8 hours, about 55 wt% to about 85 wt% over about 0 to about 14 hours, and about 80 wt% to about 100 wt% over about 0 to about 24 hours. In a particular embodiment, a sustained-release oral dosage form can release compounds of formula (I) and / or formula (II) from the dosage form in delivery patterns of approximately 0 wt% to approximately 20 wt% over approximately 0 to approximately 4 hours, approximately 20 wt% to approximately 50 wt% over approximately 0 to approximately 8 hours, approximately 55 wt% to approximately 85 wt% over approximately 0 to approximately 14 hours, and approximately 80 wt% to approximately 100 wt% over approximately 0 to approximately 20 hours. In a particular embodiment, a sustained-release oral dosage form can release compounds of the present invention from the dosage form in delivery patterns of approximately 0 wt% to approximately 20 wt% over approximately 0 to approximately 2 hours, approximately 20 wt% to approximately 50 wt% over approximately 0 to approximately 4 hours, approximately 55 wt% to approximately 85 wt% over approximately 0 to approximately 7 hours, and approximately 80 wt% to approximately 100 wt% over approximately 0 to approximately 8 hours.

[0195] The sustained-release oral dosage form containing the creatine prodrug compound of the present invention can provide creatine at concentrations present in the patient's plasma, blood, or tissues over time after oral administration. The concentration characteristics of the creatine can exhibit an AUC proportional to the dose of the corresponding compound of the present invention.

[0196] Regardless of the specific form of the controlled-release oral dosage form used, the compounds of the present invention can be released from the orally administered dosage form for a sufficient period of time to provide the patient's plasma and / or blood with long-lasting therapeutic concentrations of the compounds of the present invention. After oral administration, the dosage form containing the compounds of the present invention can provide the patient's plasma and / or blood with therapeutically effective concentrations of creatine for a continuous period of time at least about 4 hours, at least about 8 hours, at least about 12 hours, at least about 16 hours, and in certain embodiments, at least about 20 hours after oral administration of the dosage form to the patient. The continuous period of time for which therapeutically effective concentrations of creatine are maintained can be the same or different. The continuous period for which therapeutically effective plasma concentrations of creatine are maintained can begin immediately after oral administration or after a certain time interval.

[0197] In a particular embodiment, oral administration for treating a patient's disease, disorder, or symptoms The drug may contain the compound of the present invention, and the oral dosage form is adapted to provide a therapeutically effective concentration of creatine in the patient's plasma for a first consecutive period of time selected from at least about 4 hours, at least about 8 hours, at least about 12 hours, at least about 16 hours, and at least about 20 hours, following a single administration of the oral dosage form to the patient.

[0198] How to use The creatine kinase (creatine-creatine phosphate) system plays multiple roles in maintaining intracellular energy homeostasis (see, for example, Walsh et al., J Physiol, 2001, 537, 971-978). Phosphocreatine acts as a transient energy buffer in intracellular high-energy transport sites, where the rate of ATP utilization is greater than the rate of ATP production by mitochondrial respiration. Mitochondrial creatine kinase transfers the high-energy phosphate bond of newly synthesized ATP to creatine, thereby producing phosphocreatine, which is far more stable than ATP. Phosphocreatine can diffuse throughout the cell, and its high-energy phosphate bond can be used to regenerate ATP from ADP in high-energy utilization sites where other creatine kinase enzymes are strategically positioned. Such sites include membranes involved in ion transport, axonal regions involved in the transport of substances along microtubules to or from presynaptic terminals, and presynaptic terminals where energy is required for neurotransmission. Neurons synthesize creatine, however, creatine levels can be severely depleted during injury. Similar to the skeletal and cardiac muscle, neuronal creatine stores can be increased to some extent by oral creatine supplementation. The creatine kinase system also functions as an intracellular spatial energy transport mechanism. In this role as an energy carrier, energy produced by the mitochondrial ATP-ADP system is coupled to the cytosolic creatine-creatine phosphate system, which then couples to the extramitochondrial ATP-ADP system at sites of high intracellular energy transfer. The creatine-creatine phosphate system is also thought to function as a low-threshold ADP sensor that maintains the intracellular ATP-ADP concentration ratio, in which case creatine kinase is functionally coupled to ATP consumption and ATP production pathways.For example, it has been shown that creatine can react with ATP derived from mitochondrial respiration in a reaction catalyzed by mitochondrial creatine kinase, and functionally couple with adenine nucleotide translocase, thereby leading to an increase in local ADP concentration and stimulation of mitochondrial respiration. Therefore, the creatine kinase system is particularly important for maintaining energy homeostasis, including ATP homeostasis, in cells, tissues, and organs that require high energy consumption, such as neurons and muscles.

[0199] The compounds and pharmaceutical compositions of the present invention may be useful in treating diseases, disorders, or symptoms in patients associated with dysfunction of energy metabolism. In certain embodiments, dysfunction of energy metabolism includes depletion of intracellular ATP concentration, decreased intracellular creatine phosphate concentration, decreased intracellular creatine phosphate to ATP concentration ratio, and / or dysfunction of the creatine kinase system in diseased tissue or organ. In certain embodiments, dysfunction of energy metabolism includes decreased intracellular ATP concentration in diseased tissue or organ. In certain embodiments, dysfunction of energy metabolism includes decreased intracellular creatine phosphate concentration in diseased tissue or organ. In certain embodiments, dysfunction of energy metabolism includes dysfunction of the creatine kinase system and / or other intracellular energy pathways in diseased tissue or organ. In certain embodiments, diseases associated with dysfunction of energy metabolism are selected from ischemia, oxidative stress, neurodegenerative diseases, ischemia-reperfusion injury, cardiovascular diseases, multiple sclerosis, psychotic disorders, and muscle fatigue. In certain embodiments, treatment of the disease includes restoring energy homeostasis to the diseased tissue or organ.

[0200] The compounds of the present invention and their pharmaceutical compositions can be used to treat patient diseases associated with oxidative stress by administering a therapeutically effective amount of the compounds or their pharmaceutical compositions to a patient in need of such treatment. In certain embodiments, oxidative stress is associated with ischemia or neurodegenerative diseases. The method of the present invention involves treating oxidatively stressed tissue or organ by contacting the tissue or organ with the compounds or their pharmaceutical compositions.

[0201] The compounds and pharmaceutical compositions of the present invention may be useful in treating diseases, disorders, or symptoms in which a rapid increase in intracellular creatine levels has a therapeutic effect.

[0202] ischemia The compounds and pharmaceutical compositions of the present invention can be used to treat acute or chronic ischemic diseases, disorders, or symptoms. Ischemia is an imbalance between oxygen supply and demand in a cell, tissue, or organ. Ischemia is characterized by hypoxia, including anoxia, a deficiency of metabolites for normal cellular bioenergy dynamics, and the accumulation of metabolic waste products. Ischemia of a tissue or organ may be caused by circulatory insufficiency, e.g., arteriosclerosis, thrombosis, embolus formation, torsion or compression, hypotension such as shock or hemorrhage, enlargement of tissue masses (hypertrophy), increased workload (tachycardia, exercise load), and / or decreased tissue stress such as cardiac dilation. Ischemia can also occur as a result of trauma or surgical procedures. Depending on the severity and duration of the injury, ischemia may lead to a reversible decrease in cellular function or irreversible cell death. The threshold for ischemic injury varies by cell type, and its value depends, at least in part, on the cellular energy requirements of the affected tissue or organ. Parenchymal cells such as neurons (3-4 min), cardiomyocytes, hepatocytes, renal tubular cells, gastrointestinal epithelium (20-80 min) and fibroblasts, epidermis, and skeletal muscle (several hours) are more susceptible to ischemic injury than interstitial cells. Multiple studies have suggested a correlation between the functional capacity of the creatine kinase system and the ischemic tolerance of given tissues, and have indicated that strategies to improve the functional capacity of the creatine kinase system may be effective in improving the ischemic tolerance of tissues (see, for example, Wyss and Kaddurah-Daouk, Physiological Reviews, 2000, 80(3), 1107-1213, which is incorporated herein by reference in its entirety). For example, oral creatine supplementation inhibits mitochondrial cytochrome C release and downstream caspase 3 activation, resulting in ischemic neuroprotection. In connection with the inhibition of cytochrome C release and caspase 3 activation and neuroprotection, creatine administration inhibits ischemia-mediated ATP depletion.

[0203] The compounds and pharmaceutical compositions of the present invention can be used to treat acute or chronic ischemia. In certain embodiments, the compounds or compositions may be particularly useful for the acute or emergency treatment of ischemia of tissues or organs characterized by high energy demands, such as the brain, nerves, heart, lungs, kidneys, or intestines.

[0204] Due to its high energy requirements relative to its low energy storage, the brain is particularly vulnerable to hypoxia. Although the brain accounts for only a small percentage of total body weight (about 2%), it accounts for a disproportionately high percentage of oxygen consumption (about 20%). Under physiological conditions, when oxygen demand increases, this is quickly and adequately compensated for by increased cerebral blood flow. The longer the period of hypoxia / ischemia, the larger and more diffuse the affected brain area becomes. The areas most vulnerable to ischemic injury are the brainstem, hippocampus, and cerebral cortex. Unless oxygen supply is restored, the injury progresses and eventually becomes irreversible. Acute cell death occurs mainly through necrosis, but hypoxia also triggers delayed apoptosis. In addition, glutamate release from presynaptic neurons leads to Ca 2+ This could further increase the inflow and potentially lead to catastrophic collapse of postsynaptic cells. If the ischemia is not so severe, the cells will be called borderline. The reperfusion process can suppress several functions, namely protein synthesis and spontaneous electrical activity, and these suppressed functions can be restored when the O2 supply is resumed. However, the process of restoring oxygen levels in ischemic-stressed tissue, such as reperfusion, can also induce irreversible cell death, mainly through the generation of reactive oxygen species and inflammatory cell infiltration.

[0205] Neurons have limited energy-producing substances available to them, primarily glucose, ketones, or lactate. Since neurons neither produce nor store glucose nor ketones, they cannot survive for any length of time without substances absorbed and used directly or indirectly from the bloodstream. That is, a constant supply of energy-producing substances must be constantly present in the bloodstream in sufficient quantities to supply energy to the entire brain and other parts of the body. Brain cells require glucose (or its equivalent) at a concentration of approximately 5 mM to maintain the brain's optimal rate of oxidative phosphorylation, which produces ATP. Nutrients enter cells by crossing the cell membrane. Nutrient delivery often relies on extracellular mechanisms, such as oral intake, absorption, circulatory transport, and interstitial flow. Once localized near the cell, membrane-specific processes play a role in nutrient transport, which then crosses the blood-brain barrier and enters the cell and various intracellular organelles. Nutrient transport is made possible by the breakdown of ATP by ATPases. + / K + Na produced by ATPase + Gradient can be used by cells to transport nutrient molecules across the cell membrane.

[0206] A lack of oxygen or glucose inhibits or limits the neuronal capacity for ATP synthesis. The intracellular creatine / phosphocreatine system can compensate to some extent for oxygen or glucose deficiency. Creatine kinase catalyzes the synthesis of phosphocreatine from creatine in normal brain tissue. Under ATP-depleted conditions, phosphocreatine can resynthesize ATP by donating its phosphate group to ADP. However, neuronal phosphocreatine content is limited, and after complete anoxic or ischemia, phosphocreatine is also quickly depleted. ATP depletion leads to Na + / K + It is thought that blocking ATPase causes neuronal depolarization and loss of membrane potential.

[0207] Depleted oxygen levels have several consequences for cellular bioenergy and function, which can ultimately lead to cell death. For example, dysfunctional bioenergy includes impaired calcium homeostasis. Calcium regulation plays a central role in the proper functioning and survival of neurons. Calcium pumps, located in the cell membrane, use ATP to transport calcium ions out of the neuron. Proper activity of calcium pumps is essential for maintaining homeostasis in neurons, mitochondria, and the endoplasmic reticulum. Alterations in calcium pump function regulate intracellular enzyme activity and also play a crucial role in initiating mitochondrial permeability transition, which can lead to cell death. For example, intracellular Ca 2+ Metabolism is thought to contribute to cell death in Alzheimer's disease. For example, under oxidative stress conditions, the generation of oxygen free radicals overwhelms endogenous free radical protection mechanisms. This impairs neuronal metabolism and function through direct free radical damage to key cellular biomolecules, including membrane lipids, nucleic acids, and functional proteins; as well as modulation of important signaling pathways. Neuronal function depends on the transmission of electrical impulses between cells. This activity depends on the rigorous action of several membrane proteins suspended in the phospholipid bilayer. The optimal activity of this dynamic membrane microenvironment depends on the precise state and chemical composition of the lipid components. In the absence of a suitable phospholipid environment, cellular channel proteins, enzymes, and receptors cannot sustainably achieve optimal functional levels. Furthermore, oxidative stress and / or abnormal methyl metabolism can reduce the fluidity of the membrane lipid bilayer, subsequently adversely affecting embedded functional proteins. Dysfunctional bioenergy dynamics can also adversely affect the pathways of high-energy electrons along the respiratory chain.

[0208] Apoptosis is an energy-demanding process of programmed cell death, in which individual nerve cells initiate a process that leads to cell death under appropriate conditions. Some of the mechanisms described above can initiate the apoptotic pathway, including oxidative stress, calcium overload, cellular energy deficiency, trophic factor depletion, and abnormal amyloid precursor protein processing. In ischemia, neurons in the most severely affected brain tissue areas die rapidly by necrosis, while neurons exposed to less severe hypoxia die by apoptosis. The transition from cell necrosis to apoptotic cell death is associated with an increase in intracellular ATP levels. Creatine supplementation has been shown to enhance the ability to buffer ATP levels, reduce cell death, and thereby provide protection from anaerobic and ischemic injury (Balestrino et al., Amino Acids, 2002, 23, 221-229; and Zhu et al., J Neurosci 2004, 24(26), 5909-5912, which are incorporated herein by reference in their entirety).

[0209] In certain embodiments, the compounds and pharmaceutical compositions of the present invention can be used to treat cardiovascular diseases, including cerebral ischemia (stroke) and myocardial ischemia (heart infarction). Ischemic heart disease is a leading cause of morbidity and mortality in all developed countries, as it underlies many cases of acute myocardial infarction, congestive heart failure, arrhythmias, and sudden cardiac death. In the United States, ischemic heart disease accounts for nearly 20% of all deaths (approximately 600,000 deaths annually), many of which occur before patients reach hospitals. An estimated 1.1 million Americans experience a new or recurrent acute myocardial infarction each year, with many survivors continuing to experience the disease, leading to heart failure and death. As the population ages and co-occurring conditions such as obesity and diabetes become more prevalent, the public health burden caused by ischemic heart disease appears to be increasing.

[0210] Optimal cellular bioenergy dynamics depend on: (1) adequate delivery of oxygen and substrates to mitochondria; (2) mitochondrial oxidative capacity; (3) appropriate levels of high-energy phosphates and creatine phosphate / ATP ratio; (4) efficient energy transfer from mitochondria to energy-using sites; (5) appropriate local regulation of the ATP / ADP ratio near ATPases; and (6) effective feedback signaling from the usage site to maintain cellular energy homeostasis. Deficiencies in these cardiac energy pathways have been found in cardiovascular diseases, such as dilated and hypertrophic cardiomyopathy of various causes, cardiac conduction disorders, and ischemic heart disease (Saks et al., J Physiol 2006, 571.2, 253-273; Ventura-Clapier et al., J Physiol 2003, 555.1,1-13; and Ingwall and Weiss, Circ Res 2004, 95, 135-145, which are respectively incorporated herein by reference in their entirety). Decreased creatine phosphate / ATP ratios have been consistently reported in human and experimental heart failure, even under moderate workload. Creatine, creatine transporters, creatine phosphate, and ATP are significantly reduced, and a decreased creatine phosphate / ATP ratio is a predictor of mortality in congenital heart failure. Furthermore, downregulation of creatine transporter protein expression has been demonstrated in experimental animal models of heart disease and in human myocardial failure, and the generally reduced creatine phosphate and creatine levels measured in heart failure are associated with downregulated creatine transporter volume.

[0211] Cardiovascular diseases include hypertension, heart failure (e.g., congestive heart failure or heart failure after myocardial infarction), arrhythmias, diastolic disorders (e.g., left ventricular diastolic disorder), diastolic heart failure or diastolic filling disorder, systolic dysfunction, ischemia (e.g., myocardial ischemia), cardiomyopathy (e.g., hypertrophic cardiomyopathy and dilated cardiomyopathy), sudden cardiac death, myocardial fibrosis, vascular fibrosis, impaired arterial compliance, myocardial necrotizing lesions, cardiac vascular injury, cardiac vascular inflammation, post-acute myocardial infarction symptoms, and chronic post-myocardial infarction symptoms. These include myocardial infarction (including both phenotypes), coronary angioplasty, left ventricular hypertrophy, decreased ejection fraction, coronary thrombosis, cardiac lesions, cardiac vascular wall hypertrophy, vascular endothelial thickening, myocarditis, and coronary artery disease, such as fibrinoid necrosis or coronary artery disease. Ventricular hypertrophy due to systemic hypertension associated with coronary ischemic heart disease is considered a major risk factor for sudden death, post-infarction heart failure, and cardiac rupture. Patients with severe left ventricular hypertrophy are particularly susceptible to hypoxia or ischemia.

[0212] The neuroprotective effect of the compounds of the present invention was demonstrated in animal models of cerebral ischemia, e.g., Cimino et al. al., Neurotoxicol 2005, 26(5), 9929-33; Konstas et al., Neurocrit Care 2006, 4(2), 168-78; Wasterlain et al., Neurology This can be confirmed using references such as those found in 1993, 43(11), 2303-10 and Zhu et al., J Neuroscience 2004, 24(26), 5909-5912.

[0213] ischemia reperfusion injury Reperfusion injury is damage to tissue that occurs when blood supply is restored to the tissue after a period of ischemia. The lack of oxygen and nutrients from the blood to the tissue or organ creates a condition where the restoration of circulation results in oxygen-induced inflammation and oxidative damage rather than a restoration of normal function. The damage in ischemia-reperfusion injury is partly due to the inflammatory response of the damaged tissue. Reperfusion is a contributing factor to the ischemic cascade of the brain associated with stroke and traumatic brain injury. Repeated episodes of ischemia and reperfusion also appear to be a contributing factor to the formation and unsuccessful healing of chronic wounds such as bedsores and diabetic foot ulcers (Mustoe, Am J Surgery 2004, 187(5), S65-S70, which is incorporated herein by reference in its entirety). In certain embodiments, the methods and compositions of the present disclosure can protect muscles and organs, such as the heart, liver, kidneys, brain, lungs, spleen, and steroid-producing organs, such as the thyroid gland, adrenal glands, and gonads, from damage resulting from ischemia-reperfusion injury.

[0214] Ischemia and subsequent reperfusion are the leading causes of skeletal and myocardial injury in mammals. Ischemia is caused by reduced oxygen supply to tissues or organs as a result of decreased blood flow, which can lead to organ dysfunction. Reduced blood supply can result from vascular thrombosis, such as occlusion or diversion due to myocardial infarction, stenosis, accidental vascular injury, or surgical procedures. Subsequent reconstruction of adequate oxygen-containing blood supply to tissues or organs can result in increased damage, a process known as ischemia-reperfusion injury or occlusion-reperfusion injury. Complications resulting from ischemia-reperfusion injury include stroke, fatal or non-fatal myocardial infarction, myocardial remodeling, aneurysms, peripheral vascular disease, tissue necrosis, renal failure, and postoperative muscle tone reduction.

[0215] The restoration of coronary blood flow after ischemia in a transient period (reperfusion), while necessary for muscle cell survival and restoration of aerobic metabolism, introduces a set of independent stresses that can exacerbate cellular damage. Reactive oxygen species generated during reperfusion can damage proteins and membrane structures within cardiomyocytes and activate signaling pathways that lead to apoptosis. Leukocyte adhesion to endothelial cells after ischemia can clog capillaries and release inflammatory mediators. During reperfusion, the inward influx of activated components, catecholamines, and other signaling molecules contained in the plasma or locally produced within the myocardial wall may also affect the process of intracellular events in the cardiomyocytes. As a direct consequence of ischemia, reperfusion injury is a significant feature of acute coronary syndrome. Such injury can occur both spontaneously as a consequence of fibrinolysis in coronary thrombosis and as a consequence of fibrinolytics in acute angioplasty, a treatment commonly used today to open occluded vessels.

[0216] In certain embodiments, the compounds and compositions thereof of the present invention can be used to treat or reduce symptoms associated with ischemia-reperfusion injury. Ischemia-reperfusion injury may be associated with oxygen deficiency, neutrophil activation, and / or myeloperoxidase production. Ischemia-reperfusion injury may be the result of multiple disease conditions or may be iatrogenically induced, such as by thrombosis, stenosis, or surgery.

[0217] In certain embodiments, the compounds and compositions thereof of the present invention can be used to treat stroke, fatal or non-fatal myocardial infarction, peripheral vascular disease, tissue necrosis, and renal failure resulting from ischemia-reperfusion injury, as well as postoperative muscle tone reduction. In certain embodiments, the methods and compositions of the present invention reduce or mitigate the degree of ischemia-reperfusion injury.

[0218] In certain embodiments, the compounds and compositions thereof of the present invention can be used to treat, reduce, or prevent ischemia-reperfusion injury associated with vascular stenosis, thrombosis, accidental vascular injury, or occlusion or diversion of blood vessels by surgical procedures.

[0219] In certain embodiments, the compounds and compositions thereof of the present invention can also be used to treat any other conditions associated with ischemia-reperfusion, such as myocardial infarction, stroke, intermittent claudication, peripheral artery disease, acute coronary syndrome, cardiovascular disease, and muscle injury resulting from vascular occlusion.

[0220] In certain embodiments, the compounds and compositions thereof of the present invention can be used to treat myocardial infarction, stenosis, at least one blood clot, stroke, intermittent claudication, peripheral artery disease, acute coronary syndrome, cardiovascular disease, or reperfusion injury associated with muscle injury as a result of vascular occlusion.

[0221] In certain embodiments, the compounds and compositions thereof of the present invention can be used in conjunction with cardiac surgery, for example, in addition to or with cardiac arrest fluid, to prevent or minimize myocardial ischemia or reperfusion injury. In certain embodiments, the method and compositions can be used in conjunction with a cardiopulmonary bypass device during cardiac surgery to prevent or reduce myocardial ischemia-reperfusion injury.

[0222] In certain embodiments, the methods and compositions of the present invention can protect muscles and organs, such as the heart, liver, kidneys, brain, lungs, spleen, and steroid-producing organs, such as the thyroid gland, adrenal glands, and gonads, from damage resulting from ischemia-reperfusion injury.

[0223] The compounds and pharmaceutical compositions of the present invention can be used to treat ischemia-reperfusion injury of a tissue or organ by contacting that tissue or organ with an effective amount of the compound or pharmaceutical composition. The tissue or organ may be located within the patient or outside the patient, i.e., outside the body. The tissue or organ may also be a transplanted tissue or organ, and the compound or pharmaceutical composition can be contacted with the transplanted tissue or organ before removal, during transport, during transplantation, and / or after the tissue or organ has been transplanted into the recipient.

[0224] In certain embodiments, the compounds or pharmaceutical compositions of the present invention can be used to treat ischemia-reperfusion injury caused by surgery, such as cardiac surgery. The compounds or pharmaceutical compositions can be administered before, during, and / or after surgery. In certain embodiments, the compounds or pharmaceutical compositions of the present invention can be used to treat ischemia-reperfusion injury of muscle, including myocardium, skeletal muscle, or smooth muscle, and in certain embodiments, to treat ischemia-reperfusion injury of organs, such as the heart, lungs, kidneys, spleen, liver, neurons, or brain. It can be used for therapeutic purposes. The compounds of the present invention or their pharmaceutical compositions can be administered before, during, and / or after surgery.

[0225] In certain embodiments, the compounds or pharmaceutical compositions of the present invention can be used to treat ischemia-reperfusion injury of muscles, including cardiac muscle, skeletal muscle, and smooth muscle.

[0226] The efficacy of the compounds of the present invention in treating ischemia-reperfusion injury can be assessed using animal models and in clinical trials. Examples of useful methods for assessing efficacy in treating ischemia-reperfusion injury include, for example, Prass et al., J Cereb Blood Flow Metab 2007, 27(3), 452-459; Arya et al., Life Sci 2006, 79(1), 38-44; Lee et al. This is described in al., Eur. J. Pharmacol 2005, 523(1-3), 101-108; and U.S. Patent Application No. 2004 / 0038891. Useful methods for evaluating transplant perfusion / reperfusion are described, for example, in Ross et al., Am J. Physiol-Lung Cellular Mol. Physiol. 2000, 279(3), L528-536.

[0227] transplant perfusion In certain embodiments, the compounds of the present invention or their pharmaceutical compositions can be used to enhance organ transplant viability by perfusing organs with the compounds of the present invention or their pharmaceutical compositions. An increase in creatine phosphate levels is expected to prevent or minimize ischemic damage to organs. Perfusion with creatine prodrugs during organ retrieval, after donor organ retrieval, during transplantation, and / or after organ transplantation can improve the viability of organs, particularly metabolically active organs such as the heart or pancreas, thereby reducing rejection rates and / or extending the time window for organ transplantation.

[0228] In certain embodiments, the compounds and compositions thereof of the present invention can be used to treat, prevent, or reduce ischemia-reperfusion injury in extracorporeal tissues or organs. Extracorporeal tissues or organs are tissues or organs that are not in an organism (also referred to as ex vivo), such as those in transplantation. In tissue and organ transplantation, the extracted donor tissues and organs are also susceptible to reperfusion injury during extraction, transport, transplantation, and after transplantation to the recipient. The methods and compositions can be used, for example, to enhance the viability of transplanted tissues or organs by supplementing solutions used to maintain or preserve them. For example, the methods and compositions can be used to immerse transplanted tissues or organs during transport, or to keep them in contact with transplanted tissues or organs before, during, or after transplantation.

[0229] Neurodegenerative diseases Neurodegenerative diseases characterized by cell death can be classified into acute diseases such as stroke, traumatic brain injury, and spinal cord injury, as well as chronic diseases such as amyotrophic lateral sclerosis, Huntington's disease, Parkinson's disease, and Alzheimer's disease. Although these diseases have diverse causes and affect various neuronal populations, they share similar impairments in intracellular energy metabolism. For example, intracellular ATP levels decrease, and Ca2+ levels decrease. 2+ This leads to cytoplasmic accumulation and stimulation of the formation of oxygen species. Ca 2+ Reactive oxygen species can then induce apoptotic cell death. Regarding these impairments, impaired cerebral creatine metabolism is also evident, as it is reflected in decreased total creatine concentration, creatine phosphate concentration, creatine kinase activity, and / or creatine transporter content (e.g., Wyss and Kaddurah-Daouk, Physiol Rev 2000, 80, 1107-1213; Tarnopolsky and Beal, Ann Neuro). See l 2001, 49, 561–574; and Butterfield and Kanski, Mech Ageing Dev 2001, 122, 945–962 (these are respectively incorporated herein by reference in their entirety).

[0230] Acute and chronic neurodegenerative diseases are associated with high morbidity and mortality rates, and there are few treatment options available. Many neurodegenerative diseases, including stroke, traumatic brain injury, spinal cord injury, amyotrophic lateral sclerosis (ALS), Huntington's disease, Alzheimer's disease, and Parkinson's disease, are characterized by neuronal cell death. Cell death occurs through necrosis or apoptosis. Necrotizing cell death in the central nervous system follows acute ischemia or traumatic injury to the brain or spine. This cell death occurs in the areas most severely affected by rapid biochemical breakdown, which leads to the generation of free radicals and excitotoxins. This is followed by mitochondrial and nuclear swelling, organelle degradation, and chromatin condensation around the nucleus, as well as rupture of the nuclear and cytoplasmic membranes and DNA degradation through disorderly enzymatic cleavage. Apoptotic cell death can be characteristic of both acute and chronic neurological diseases. Apoptosis occurs in areas not severely affected by the injury. For example, after ischemia, necrotizing cell death occurs in the core of the lesion where hypoxia is most severe, while apoptosis occurs in the border zone where collateral circulation reduces the degree of hypoxia. Apoptotic cell death is also a component of lesions that appear after brain or spinal cord injury. In chronic neurodegenerative diseases, apoptosis is the primary form of cell death. In apoptosis, a biochemical cascade activates proteases that disrupt molecules necessary for cell survival and other enzymes that mediate the cell death program. Caspases contribute directly or indirectly to morphological changes in cells during apoptosis (Friedlander, N Engl J Med 2003, 348(14)). 1365-75). Oral creatine supplementation has been shown to inhibit mitochondrial cytochrome C release and downstream caspase-3 activation in the caspase-mediated cell death cascade during cerebral ischemia, as well as inhibiting ATP depletion (Zhu et al., J Neurosci 2004, 24(26), 5909-5912), suggesting that manipulating the creatine kinase system may be effective in controlling apoptotic cell death in chronic neurodegenerative diseases.

[0231] Creatine administration has demonstrated neuroprotective effects, particularly in animal models of Parkinson's disease, Huntington's disease, and ALS (Wyss and Schulze, Neuroscience 2002, 112(2), 243-260, which is incorporated herein by reference in its entirety), suggesting that oxidative stress levels may be a determinant of metabolism in various neurodegenerative diseases. Current hypotheses regarding the mechanisms of creatine-mediated neuroprotection include improved energy storage and stabilization of mitochondrial membrane permeability transition pores by the octameric structure of creatine kinase. Therefore, higher intracellular creatine levels are thought to improve the overall bioenergy state of cells and increase their resistance to injury.

[0232] Parkinson's disease Parkinson's disease is a slowly progressive degenerative disorder of the nervous system characterized by resting tremor, bradykinesia (slowness of voluntary movement), and increased muscle tone (rigidity). In Parkinson's disease, nerve cells in the basal ganglia, such as those in the substantia nigra, degenerate, leading to a decrease in dopamine production and the number of connections between nerve cells in the basal ganglia. As a result, the basal ganglia lose the ability to coordinate smooth muscle movement and postural changes, leading to tremor, incoordination, and slow, reduced movement (bradykinesia) (Blandini, et al., Mol. Neurobiol). (1996, 12, 73-94).

[0233] Oxidative stress is thought to be a possible factor in metabolic degradation observed in Parkinson's disease tissue (Ebadi et al., Prog Neurobiol 1996, 48, 1-19; Jenner and Olanow, Ann Neurol). Creatine supplementation has been shown to exert neuroprotective effects (Matthews et al., Exp Neurol, 1999, 157, 142-149, both incorporated herein by reference in their entirety).

[0234] The efficacy of administering the compounds of the present invention for the treatment of Parkinson's disease can be assessed in animal and human models of Parkinson's disease, as well as in clinical trials. Known animal and human models of Parkinson's disease are available (e.g., O'Neil et al., CNS Drug Rev. 2005, 11(1), 77-96; Faulkner et al., Ann. Pharmacother. 2003, 37(2), 282-6; Olson et al., Am. J. Med. 1997, 102(1), 60-6; ​​Van Blercom et al., Clin Neuropharmacol. 2004, 27(3), 124-8; Cho et al., Biochem. Biophys. Res. Commun. 2006, 341, 6-12; Emborg, J.Neuro. Meth. 2004, 139, 121-143; Tolwani et al., Lab Anim Sci 1999, 49(4), 363-71; Hirsch et al., J Neural Transm Suppl 2003, 65, 89-100; Orth and Tabrizi, Mov Disord 2003, 18(7), 729-37; Betarbet et al., Bioessays 2002, 24(4), 308-18; and McGeer and McGeer, Neurobiol Aging 2007, 28(5), 639-647).

[0235] Alzheimer's disease Alzheimer's disease is a progressive loss of mental function characterized by the degeneration of brain tissue, including a decrease in nerve cells and the development of senile plaques and neurofibrillary groves. In Alzheimer's disease, the brain partially degenerates, nerve cells are destroyed, and the responsiveness of maintenance neurons to neurotransmitters decreases. The abnormalities in brain tissue consist of senile plaques or neurite plaques, for example, aggregates of dead nerve cells containing an insoluble abnormal protein called amyloid, and neurofibrillary groves, twisted chains of insoluble proteins within nerve cells.

[0236] Oxidative stress may be a factor in the metabolic degradation observed in Alzheimer's disease tissue, and creatine kinase is thought to be one of the targets of oxidative damage (Pratico et al., FASEB J 1998, 12, 1777-1783; Smith). Studies have shown a correlation between intracellular levels of creatine phosphate and the progression of dementia (Pettegrew et al., Neurobiol Aging 1994, 15, 117-132, which is incorporated herein by reference in its entirety).

[0237] The efficacy of administering the compounds of the present invention for the treatment of Alzheimer's disease can be assessed in animal and human models of Alzheimer's disease, as well as in clinical trials. An animal model useful for assessing the efficacy of the compounds for the treatment of Alzheimer's disease is, for example, Van Dam and De Dyn, Nature Revs Drug Disc 2006, 5, 956-970; Simpkins et al., Ann NY Acad Sci, 2005, 1052, 233-242; Higgins and Jacobsen, Behav Pharmacol 2003, 14(5-6), 419-38; Janus and Westaway, Physiol Disclosed in Behav 2001, 73(5), 873-86; and Conn, ed., "Handbook of Models in Human Aging," 2006, Elsevier Science & Technology.

[0238] Huntington's disease Huntington's disease is an autosomal dominant neurodegenerative disorder in which specific cell death occurs in the neostriatum and cortex (Martin, N Engl J Med 1999, 340, 1970-80, which is incorporated herein by reference in its entirety). Onset typically occurs in the 40s or 50s, and the average survival time is 14–20 years from onset. Huntington's disease is fatal, and there is no effective treatment. Symptoms include characteristic motor impairment (Huntington's chorea), cognitive impairment, and psychiatric symptoms. The disease is caused by a mutation that codes for abnormal elongation of the CAG polyglutamine repeat in the huntingtin protein. Multiple studies suggest the presence of a progressive impairment of energy metabolism, which may originate from mitochondrial damage caused by oxidative stress as a consequence of free radical generation. Preclinical studies in animal models of Huntington's disease have reported the neuroprotective effects of creatine administration. For example, creatine neuroprotection is associated with elevated levels of creatine phosphate and creatine, as well as decreased lactate levels in the brain, which is consistent with improved energy production (see Ryu et al., Pharmacology & Therapeutics 2005, 108(2), 193-207, which is incorporated herein by reference in its entirety).

[0239] The efficacy of administering the compounds of the present invention for the treatment of Huntington's disease can be assessed in animal and human models of Huntington's disease, as well as in clinical trials. Examples of animal models of Huntington's disease include Riess and Hoersten, U.S. Patent Application No. 2007 / 0044162; Rubinsztein, Trends in Genetics, 2002, 18(4), 202-209; Matthews et al., J. Neuroscience 1998, 18(1), 156-63; and Tadros et al., Pharmacol Biochem Behav. Disclosed in 2005, 82(3), 574-82, and U.S. Patent No. 6,706,764, and U.S. Patent Applications No. 2002 / 0161049, No. 2004 / 0106680, and No. 2007 / 0044162. A placebo-controlled clinical trial evaluating the efficacy of creatine supplementation for the treatment of Huntington's disease is disclosed in Verbessem et al., Neurology 2003, 61, 925-230.

[0240] Amyotrophic lateral sclerosis (ALS) Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by the progressive and specific loss of motor neurons in the brain, brainstem, and spine (Rowland and Schneider, N Engl J Med 2001, 344, 1688-1700, which is incorporated herein by reference in its entirety). ALS begins with weakness, which most often occurs in the hands and less frequently in the feet, generally progressing to the arms or legs. Over time, weakness increases, and spasms characterized by muscle cramps and contractures occur, followed by muscle spasms, and sometimes tremors. The mean age of onset is 55 years, and the clinically projected life expectancy after onset is 4 years. The only approved treatment for ALS is riluzole, which can only extend survival by about 3 months. Oral creatine has been shown to provide neuroprotective effects in genetically modified animals with ALS (Klivenyi et al., Nat Med 1999). 5, 347-50 (these are incorporated herein by reference in their entirety).

[0241] The efficacy of administering the compounds of the present invention for the treatment of ALS can be assessed in animal and human models of ALS, as well as in clinical trials. Natural disease models of ALS include mouse models (motor neuronal degeneration, progressive motor neuropathy, and unsteadiness) and canine models of hereditary canine spinal muscular atrophy (Pioro and Mitsumoto, Examples include Clin Neurosci, 19954996, 3(6), 375-85). Experimentally generated and genetically engineered animal models of ALS can also be useful in assessing therapeutic efficacy (e.g., Doble and Kennelu, See Amyotroph Lateral Scler Other Motor Neuron Disord. 2000, 1(5), 301-12; Grieb, Folia Neuropathol. 2004, 42(4), 239-48; Price et al., Rev Neurol (Paris), 1997, 153(8-9), 484-95; and Klivenyi et al., Nat Med 1999, 5, 347-50). Specifically, the SOD1-G93A mouse model is recognized as an ALS model. Examples of clinical trial protocols useful for assessing the treatment of ALS are described, for example, in Mitsumoto, Amyotroph Lateral Scler Other Motor Neuron Disord. 2001, 2 Suppl 1, S10-S14; Meininger, Neurodegener Dis 2005, 2, 208-14; and Ludolph and Sperfeld, Neurodegener Dis. 2005, 2(3-4), 215-9.

[0242] Multiple sclerosis Multiple sclerosis (MS) is a multifaceted inflammatory autoimmune disease of the central nervous system caused by an autoimmune attack on the insulating axonal myelin sheets of the central nervous system. Demyelination leads to disruption of transmission, resulting in serious disease with local axonal destruction and irreversible neuronal cell death. The symptoms of MS vary greatly from patient to patient, each presenting a specific pattern of motor, sensory, and paresthesia. Pathologically, MS is characterized by multiple inflammatory lesions, demyelinating plaques, gliosis, and axonal pathology within the brain and spinal cord, all contributing to the clinical pathogenesis of neurological physical disorders (see, e.g., Wingerchuk, Lab Invest 2001, 81, 263-281; ​​and Virley, NeruoRx 2005, 2(4), 638-649). Although the causative events that trigger the disease are not fully understood, most evidence points to an autoimmune etiology, in conjunction with environmental factors and specific genetic predispositions. Functional impairment, physical disability, and handicaps manifest as motor paralysis, sensory and cognitive impairments, seizures, tremors, coordination disorders, and visual impairments, which affect the individual's quality of life. While the clinical course of MS can vary considerably from person to person, the disease can always be classified into three types: relapsing-remitting, secondary progressive, and primary progressive. Several studies link dysfunction in creatine phosphate metabolism to the etiology and symptoms of this disease (Minderhoud et al., Arch Neurol). 1992, 49(2), 161-5; He et al., Radiology 2005, 234(1), 211-7; Tartaglia et al., Although creatine supplementation alone does not appear to be effective in improving exercise capacity in individuals with MS (Lambert et al., Arch Phys Med Rehab 2003, 84(8), 1206-1210), (Arch Neurology 2004, 61(2), 201-207; Duong et al., J Neurol 2007, Apr.20; and Ju et al., Magnetic Res Imaging 2004, 22, 427-429), creatine supplementation alone does not appear to be effective in improving exercise capacity in individuals with MS.

[0243] The assessment of MS treatment efficacy in clinical trials is based on the Comprehensive Disability Assessment Scale (Kurtzke, Neurology 1983, 33, 1444-1452) and the MS Functional Composite Assessment (Fischer et al., Mult Scler, 1999, 5, 244- This can be achieved using tools such as magnetic resonance imaging lesion load, biomarkers, and self-reported quality of life (see, e.g., Kapoor, Cur Opinion Neurol 2006, 19, 255-259). Animal models of MS that have been shown to be useful for identifying and validating promising therapeutic agents include experimental autoimmune / allergic encephalomyelitis (EAE) rodent models that stimulate the clinical and pathological manifestations of MS (Werkerle and Kurschus, Drug Discovery Today: Disease Models, Nervous System Disorders, 2006, 3(4), 359-367; Gijbels et al., Neurosci Res Commun 2000, 26, 193-206; and Hofstetter et al., J Immunol 2002, 169, 117-125), as well as non-human primate EAE models ('t Hart et al., Immunol Today 2000, 21, 290-297).

[0244] Mental disorder In certain embodiments, the compounds of the present invention or their pharmaceutical compositions can be used to treat mental disorders, such as schizophrenia, bipolar disorder, and anxiety.

[0245] Schizophrenia Schizophrenia is a chronic, severe, and physically debilitating brain disorder affecting approximately 1% of the world's population, including 3.2 million Americans. Schizophrenia is a group of neuropsychiatric disorders characterized by dysfunction of thought processes, such as delusions, hallucinations, and widespread disengagement of interest in others. Schizophrenia includes subtypes such as paranoid schizophrenia, characterized by mental occupation with delusions or auditory hallucinations; hebephrenic or disorganized schizophrenia, characterized by disorganized speech, disorganized behavior, and flat or inappropriate affect; catatonic schizophrenia, where physical symptoms such as immobility, hyperkinesia, or stereotyped postures are dominant; undifferentiated schizophrenia, characterized by a combination of symptom features from other subtypes; and residual schizophrenia, where an individual does not currently have positive symptoms but exhibits negative and / or cognitive symptoms of schizophrenia (DSM-IV-TR classification 295.30 (paranoid), 295.10 (disorganized), 295.20 (catatonic), 295.90 (unadifferentiated), and 295.60 (residual); Diagnostic and Statistical Manual). of Mental Disorders, 4 th See Edition, American Psychiatric Association, 297-319, 2005. Schizophrenia includes these and other closely related mental disorders, such as schizophrenia-like disorders, schizoaffective disorders, delusional disorders, short-term mental disorders, shared psychotic disorders, mental disorders due to systemic symptoms, substance-induced mental disorders, and unspecified mental disorders (DSM-IV-TR, 4). th Edition, pp. 297-344, American Psychiatric Association, 2005).

[0246] Schizophrenia symptoms can be classified as positive, negative, or cognitive. Positive symptoms of schizophrenia include delusions and hallucinations, which can be measured, for example, using the Positive-Negative Symptom Rating Scale (PANSS) (Kay et al., Schizophrenia Bulletin 1987, 13, 261-276). Negative symptoms of schizophrenia include affective blunting, anergy, aphasia, and social withdrawal, which can be measured, for example, using the Negative Symptom Rating Scale (SANS) (Andreasen, 1983, Scales for the Assessment of Negative Symptoms (SANS), Iowa City, Iowa). Cognitive symptoms of schizophrenia include impairments in organizing and cognitive use, which can be measured using the Positive-Negative Symptom Rating Scale-Cognitive Subscale (PANSS-Cognitive Subscale) (Lindenmayer et al., J Nerv Ment Dis 1 Using 994, 182, 631-638) or by assessing the ability to perform cognitive tasks, for example, the Wisconsin Card Sorting Test (e.g., Green This can be measured using the following methods (see et al., Am J Psychiatry 1992, 149, 162-67; and Koren et al., Schizophr Bull 2006, 32(2), 310-26).

[0247] Multiple studies support a correlation between schizophrenia and dysfunction in high-energy phosphate metabolism in the brain (Fukuzako, World J Biol Psychiatry). (2001, 2(2), 70-82; and Gangadhar et al., Prog Neuro-Psychopharmacology & Biological Psychiatry 2006, 30, 910-913). Patients with schizophrenia show reduced phosphocreatine levels in the left and right frontal regions of the brain, which are highly correlated with hostility-distrust and anxiety-depression rating subscales (Deicken et al., Biol Psychiatry 1994, 36(8), 503-510; Volz et al., Biol Psychiatry 1998, 44, 399-404; and Volz et al., Biol Psychiatry 2000, 47, 954-961). Therefore, creatine supplementation has been proposed for the treatment of schizophrenia (see, for example, Lyoo et al., Psychiatry Res: Neuroimaging 2003, 123, 87-100).

[0248] The efficacy of creatine prodrugs and their pharmaceutical compositions for the treatment of schizophrenia can be determined by methods known to those skilled in the art. For example, negative, positive, and / or cognitive symptoms of schizophrenia can be measured before and after treatment of the patient. A reduction in such symptoms indicates an improvement in the patient's symptoms. Improvement in schizophrenia symptoms can be measured, for example, using the Negative Symptom Rating Scale (SANS), the Positive and Negative Symptom Rating Scale (PANSS) (e.g., Andreasen, 1983, Scales for the Assessment of Negative Symptoms (SANS), Iowa City, Iowa; and Kay et al., Schizophrenia). It can be assessed using cognitive deficit tests, such as the Wisconsin Card Sorting Test (WCST) and other measures of cognitive function (see, for example, Keshavan et al., Schizophr Res 2004, 70(2-3), 187-194; Rush, Handbook of Psychiatric Measures, American Psychiatric Publishing 2000; Sajatovic and Ramirez, Rating Scales in Mental Health, 2nd ed, Lexi-Comp, 2003; Keefe, et al., Schizophr Res. 2004, 68(2-3), 283-97; and Keefe et al., Neuropsychopharmacology, 19 Apr. 2006).

[0249] The efficacy of creatine prodrugs and their pharmaceutical compositions can be evaluated using animal models of schizophrenic disorder (see, for example, Geyer and Moghaddam, in "Neuropsychopharmacology," Davis et al., Ed., Chapter 50, 689-701, American College of Neuropsychopharmacology, 2002). For example, conditioned avoidance response (CAR) and catalepsy tests in rats have been shown to be useful in predicting the trend of schizophrenia treatment activity and EPS effects, respectively (Wadenberg et al., Neuropsychopharmacology, 2001, 25, 633-641).

[0250] Bipolar disorder Bipolar disorder is a psychiatric disorder characterized by periods of extreme mood swings. These mood swings can range from depressive episodes (e.g., persistent feelings of sadness, anxiety, guilt, anger, loneliness, and / or despair; sleep and eating disorders; fatigue and loss of interest in activities normally enjoyed; inability to concentrate; feelings of loneliness, self-loathing, apathy or indifference; depersonalization; loss of interest in sexual activity; shyness or social anxiety; irritability; chronic pain; apathy; and morbid / suicidal ideation) to manic episodes (e.g., euphoria, excitement, and / or paranoia). Bipolar disorder is described in the Diagnostic and Statistical Manual of Mental Disorders, 4. th Bipolar disorder is identified and classified in Ed., Text Revision (DSM-IV-TR), American Psychiatric Assoc., 200, pages 382-401. Bipolar disorder includes type I bipolar disorder, type II bipolar disorder, cyclothymic disorder, and other unspecified bipolar disorders.

[0251] Patients with bipolar depression have been shown to have impaired high-energy phosphate metabolism in the brain, characterized by decreased levels of phosphocreatine and creatine kinase (Kato et al., J Affect Disord 1994, 31(2), 125-33; and Segal et al., Eur Neuropsychopharmacology 2007, 17, 194-198), likely involving mitochondrial energy metabolism (Stork and Renshaw, Molecular Psychiatry 2005, 10, 900-919).

[0252] The treatment of bipolar disorder can be assessed in clinical trials using assessment scales, such as the Montgomery-Asberg Depression Rating Scale, the Hamilton Depression Scale, the Ruskin Depression Scale, the Feiner Criteria, and / or the Global Clinical Impression Scale (Gijsman et al., Am J Psychiatry 2004, 161, 1537-1547).

[0253] anxiety Anxiety is discussed in the Diagnostic and Statistical Manual of Mental Disorders, 4 th Anxiety disorders are identified and classified in Ed., Text Revision (DSM-IV-TR), American Psychiatric Assoc., 200, pages 429-484. These include panic attacks, agoraphobia, panic disorder without agoraphobia, agoraphobia without a history of panic disorder, specific phobias, social phobias, obsessive-compulsive disorder, post-traumatic stress disorder, acute stress disorder, generalized anxiety disorder, anxiety disorder due to systemic symptoms, substance-induced anxiety disorder, and other unspecified anxiety disorders. Recent studies have described a correlation between decreased creatine / phosphocreatine levels in the center semioval (a representative area of ​​cerebral white matter) and the severity of anxiety (Coplan et al., Neuroimaging, 2006, 147). 27-39).

[0254] Useful animal models for assessing anxiety treatment include fear-conditioned startle (Brown et al., J Experimental Psychol, 1951, 41, 317-327), elevated cruciform maze (Pellow et al., J Neurosci. Methods 1985, 14, 149-167; and Hogg, Pharmacol Biochem Behavior 1996, 54(1), 21-20), and fear conditioning behavior in elevated cross mazes (Korte and De Boer, Eur J Pharmacol 2003, 463, 163-175) is an example. Genetic animal models of anxiety include other animal models that are sensitive to anxiolytics (Ma It is known as rtin, Acta Psychiatr Scand Suppl 1998, 393, 74-80 (Toh, Eur J Pharmacol) (2003, 463, 177-184).

[0255] In clinical trials, efficacy can be assessed by selecting patients using psychological procedures to induce experimental anxiety in healthy volunteers and patients with anxiety disorders (see, e.g., Graeff, et al., Brazilian J Medical Biological Res 2003, 36, 421-32), or by selecting patients based on a structured interview for DSM-IV Axis I disorders, as described in First et al., Structured Clinical Interview for DSM-IV Axis I Disorders, Patient Edition (SCIDIP), Version 2. Biometrics Research, New York State Psychiatric Institute, New York, 1995. Any of the multiple scales can be used to assess anxiety and the effectiveness of the treatment, such as the Penn State Concern Questionnaire (Behar et al., J Behar Ther Exp Psychiatr Examples include the Hamilton Anxiety and Depression Scale (2003, 34, 25-43), the Spielberger State-Trait Anxiety Inventory, and the Liebowitz Social Anxiety Scale (Hamilton, J Clin Psychiatry 1980, 41, 21-24; Spielberger and Vagg, J Personality Assess 1984, 48, 95-97; and Liebowitz, J Clin Psychiatry 1993, 51, 31-35 (previously mentioned)).

[0256] Genetic diseases affecting the creatine kinase system The intracellular creatine pool is maintained by creatine uptake from the diet and by endogenous creatine synthesis. Many tissues, especially the brain, liver, and pancreas, have Na + -Cl -This includes creatine-dependent transport (SLC6A8), which is responsible for active creatine transport across the plasma membrane. Creatine biosynthesis involves the action of two enzymes: L-arginine:glycine amidinotransferase (AGAT) and guanidinoacetate transferase (GAMT). AGAT catalyzes the transfer of the amidino group from arginine to glycine, producing ornithine and guanidinoacetate. Guanidinoacetate is then methylated at the amidino group by GAMT to yield creatine (see, for example, Wyss and Kaddurah-Daouk, Phys Rev 2000, 80, 1107-213).

[0257] In humans, two gene errors in creatine biosynthesis and one gene error in the creatine transporter are known, and these errors result in deficiencies in AGAT, GAMT, and the creatine transporter (Schulze, Cell Biochem, 2003, 244(1-2), 143-50; Sykut-Cegielska et al.). Acta Biochimica Polonica 2004, 51(4), 875-882). Patients with impaired creatine synthesis experience systemic depletion of creatine and creatine phosphate. Patients with AGAT deficiency may exhibit intellectual and motor developmental delay, severe language developmental delay, and febrile seizures (Item et al., Am J Hum Genet. 2001, 69, 1127-1133). Patients with GAMT deficiency may exhibit developmental delay without active speech, autism with self-injurious behavior, extrapyramidal symptoms, and epilepsy (Stromberger et al., J Inherit Metab Dis 2003, 26, 299-308). Patients with creatine transporter deficiency show intracellular depletion of creatine and creatine phosphate. The gene encoding the creatine transporter is located on the X chromosome, and male patients exhibit mild to severe intellectual disability, while female patients have milder symptoms (Salomons et al., J. Inherit). Metab Dis 2003, 26, 309-18; Rosenberg et al., Am J Hum Genet. 2004, 75, 97-105; deGrauw et al., Neuropediatrics 2002, 33(5), 232-238; Clark et al., Hum Genet, 2006, April).

[0258] Creatine supplementation at a dosage of approximately 350 mg to 2 g / kg body weight per day has been shown to be effective in resolving the clinical symptoms of AGAT or GAMT deficiency (see, e.g., Schulze, Cell Biochem, 2003, 244(1-2), 143-50). However, unlike patients with GAMT and AGAT deficiency, oral creatine supplementation does not result in elevated cerebral creatine levels in patients with creatine transporter deficiency (see Stockler-Ipsiroglu et al., in Physician's Guide to the Treatment and Follow up of Metabolic Diseases, eds Blau et al., Springer Verlag, 2004).

[0259] Muscle fatigue During high-intensity exercise, ATP hydrolysis is first buffered by creatine phosphate via the creatine kinase reaction (Kongas and van Beek, 2 nd Int. Conf. Systems Biol 2001, Los Angeles Calif., Omnipress, Madison, Wis., 198-207; and Walsh et al., J Physiol 2001, 537.3, 971-78 (these are each incorporated herein by reference in their entirety). During exercise, creatine phosphate is immediately available for ATP regeneration, whereas glycolysis is induced several seconds later, and the stimulation of mitochondrial oxidative phosphorylation is even more delayed. Due to the limited storage of creatine phosphate in muscles, creatine phosphate is depleted in approximately 10 seconds during high-intensity exercise. It has been suggested that muscle capacity can be improved by increasing muscle storage of creatine phosphate, thereby delaying creatine phosphate depletion. While creatine and / or creatine phosphate supplementation may improve muscle capacity at intermittent suprasmaximal exercise loads, there is no evidence that supplementation improves endurance capacity. On the other hand, intravenous injection of creatine phosphate appears to improve exercise tolerance during prolonged submaximal exercise (Clark, J Athletic Train, 1997, 32, 45-51, which is incorporated herein by reference in its entirety).

[0260] Muscle strength In normal, healthy individuals, dietary creatine supplementation has beneficial side effects on muscle function, leading to increased use of creatine by amateur and professional athletes. There is evidence suggesting that creatine supplementation may improve overall muscle capacity by increasing muscle stores of creatine phosphate, the most important energy source for immediate ATP regeneration in the first few seconds of high-intensity exercise, by accelerating the recovery of the creatine phosphate pool during the recovery phase, and by suppressing the breakdown of adenosine nucleotides and possibly lactate accumulation during exercise (see, for example, Wyss and Kaddurah-Daouk, Physiol Rev 2000, 80(3), 1107-1213).

[0261] However, in normal, healthy individuals, continuous and long-term use of creatine fails to maintain high levels of creatine and creatine phosphate in the muscles (for example, Juhn et al., Clin J Sport Med 1998, 8, 286-297; Terjung et al., Med Sci Sports Exerc See 2000, 32, 706-717; and Vandenberghe et al., J Appl Physiol 1997, 83, 2055-2063, which are respectively incorporated herein by reference in their entirety. This is likely a result of downregulation of creatine transporter activity and transporter protein content (Snow and Murphy, Mol Cell Biochem 2001, 224(1-2), 169-181 (which is incorporated herein by reference in its entirety). That is, the creatine prodrugs of the present invention can be used for maintaining, restoring, and / or improving muscle strength in mammals, particularly humans.

[0262] The efficacy of administering the compounds of the present invention for maintaining, restoring, and / or improving muscle strength can be assessed in animal models, human models, and clinical trials. Animal models available for assessing muscle strength are disclosed, for example, in Wirth et al., J Applied Physiol 2003, 95, 402-412 and Timson, J. Applied Physiol 1990, 69(6), 1935-1945. Muscle strength can be assessed in humans, for example, using the methods disclosed in Oster, U.S. Patent Application No. 2007 / 0032750 and U.S. Patent Application No. 2007 / 0012105, and / or other methods known to those skilled in the art.

[0263] organ and cell viability In certain embodiments, the isolation of living brain, muscle, pancreatic cells, or other cell types for research or cell transplantation can be improved by perfusing the cells with an isolation or growth medium containing a creatine phosphate analog prodrug and / or contacting the cells with the medium. In certain embodiments, the viability of tissue organs or cells can be improved by contacting the tissue organs or cells with an effective amount of the compound of the present invention or its pharmaceutical composition.

[0264] Diseases related to glucose level regulation Since creatine phosphate administration lowers plasma glucose levels, it may be useful in treating diseases related to glucose level regulation, such as hyperglycemia, insulin-dependent or non-insulin-dependent diabetes mellitus, and related diseases secondary to diabetes (U.S. Patent Application No. 2005 / 0256134).

[0265] The efficacy of administering the compounds of the present invention to treat diseases related to glucose level regulation can be assessed in animal models, human models, and clinical trials. The compounds can be administered to animals such as rats, rabbits, or monkeys, and plasma glucose concentrations can be measured at various time points (see, for example, U.S. Patent Application No. 2003 / 0232793). The efficacy of the compounds in treating insulin-dependent or non-insulin-dependent diabetes and related diseases secondary to diabetes can be assessed in animal models of diabetes, e.g., Shafrir, "Animal Models of Diabetes," Ed., 2007, CRC Press; Mordes et al., "Animal Models" of Diabetes," 2001, Harwood Academic Pre ss; Mathe, Diabete Metab 1995, 21(2), 106-111; and Rees and Alcolado, Diabetic Med. Evaluation can be performed using materials disclosed in 2005, 22, 359-370, etc.

[0266] dose A pharmaceutically acceptable salt or pharmaceutically acceptable solvate of the compound of the present invention or any of the above may be administered to treat diseases or disorders associated with dysfunction of energy metabolism.

[0267] The amount of the compound of the present invention effective in treating a specific disease, disorder, or symptom disclosed herein will depend on the nature of the disease, disorder, or symptom and can be determined by standard clinical techniques known in the art. In vitro or in vivo assays may optionally be employed to help identify the optimal dosage range. The amount of compound administered may depend, among other factors, on the patient being treated, the patient's weight, the patient's health, the disease being treated, the severity of the illness, the route of administration, the potency of the compound, and the judgment of the prescribing physician.

[0268] For systemic administration, the therapeutically effective dose can first be estimated from in vitro assays. For example, a certain dose can be prescribed to an animal model to obtain a beneficial circulating composition concentration range. The initial dose can also be estimated from in vivo data, e.g., animal models, using techniques known in the art. Using such information, a useful dose in humans can be determined more accurately. Those skilled in the art can optimize the administration to humans based on the animal data.

[0269] Creatine is naturally produced in the human body, with some synthesized in the kidneys, pancreas, and liver (approximately 1-2 grams per day), and some ingested through food (approximately 1-5 grams per day). Cells actively take up creatine via creatine transporters. Within cells, creatine kinase phosphorylates creatine to form a pool of creatine phosphate, which can act as a temporal and spatial energy buffer.

[0270] Creatine, creatine phosphate, and their analogues can be administered in high doses without adverse side effects. For example, creatine monohydrate has been administered to athletes and bodybuilders in doses ranging from 2 to 3 g / day, and creatine phosphate has been administered to patients with heart disease by intravenous injection up to 8 g / day without adverse side effects. Animals fed a diet containing up to 1% cyclocreatine showed no adverse effects (see, for example, Griffiths and Walker, J. Biol. Chem. 1976, 251(7), 2049-2054; Annesley et al., J Biol Chem 1978, 253(22), 8120-25; Lillie et al., Cancer Res 1993, 53, 3172-78; and Griffiths, J Biol Chem 1976, 251(7), 2049-54).

[0271] In certain embodiments, a therapeutically effective dose of the compound of the present invention may include approximately 1 mg equivalent to approximately 20,000 mg equivalent of a creatine phosphate analog per day, approximately 100 mg equivalent to approximately 12,000 mg equivalent of a creatine phosphate analog per day, approximately 1,000 mg equivalent to approximately 10,000 mg equivalent of a creatine phosphate analog per day, and, in certain embodiments, approximately 4,000 mg equivalent to approximately 8,000 mg equivalent of a creatine phosphate analog per day.

[0272] A single dose may be administered in a single dosage form or in multiple dosage forms. When multiple dosage forms are used, the amount of the compound contained in each dosage form may be the same or different. The amount of the compound of the present invention contained in a single dose may depend on the route of administration and whether the patient's disease, disorder, or symptoms are being effectively treated acutely, chronically, or in combination with acute and chronic administration.

[0273] In certain embodiments, the dose administered is less than the toxic dose. The toxicity of the compositions described herein is determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, by LD 50 (A dose that causes death in 50% of the population) or LD 100 (100% of the group will die) The dose can be determined by calculating the dose. The dose ratio between toxicity and therapeutic effect is the therapeutic index. In certain embodiments, a pharmaceutical composition may exhibit a high therapeutic index. Using data obtained from these cell culture assays and animal experiments, a range of non-toxic dosages for use in humans can be constructed. A single dose of the pharmaceutical composition of the present invention may contain an effective dose and be within a range of circulating concentrations that exhibit little to no toxicity, for example, within a range of circulating concentrations in blood, plasma, or the central nervous system. The dose may vary within this range depending on the dosage form adopted and the route of administration used.

[0274] During treatment, the dosage and administration schedule can provide an effective amount of creatine phosphate analog sufficient to treat the disease or to maintain a steady state. In certain embodiments, escalating doses may be administered.

[0275] Administration The compounds of the present invention, their pharmaceutically acceptable salts, solvates, tautomers, or stereoisomers, or pharmaceutically acceptable solvates of any of the above, or pharmaceutical compositions of any of the above, can be administered by any suitable route. In certain embodiments, the compounds of the present invention can be administered intermittently or continuously. Examples of suitable routes of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, oral, sublingual, intranasal, intracerebral, vaginal, transdermal, rectal, inhalation, or topical. Administration can be systemic or topical. Administration can be by bolus injection, continuous intravenous infusion, or by absorption through epithelial or mucocutaneous layers, such as the oral mucosa, rectum, and intestinal mucosa.

[0276] In certain embodiments, it may be desirable to introduce the compounds of the present invention, pharmaceutically acceptable salts or pharmaceutically acceptable solvates of any of the above, or pharmaceutical compositions of any of the above, directly into the central nervous system by any suitable route, including intraventricular, subarachnoid, and epidural injection. Intraventricular injection can be facilitated by using an intraventricular catheter connected to a reservoir, for example, an Onmyer reservoir.

[0277] In certain embodiments, the compounds of the present invention, their pharmaceutically acceptable salts, solvates, tautomers, or stereoisomers, or pharmaceutically acceptable solvates of any of the above, or pharmaceutical compositions of any of the above, can be administered parenterally by injection or infusion, including, for example, intravenous, intramuscular, intra-arterial, subarachnoid, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections.

[0278] The compounds of the present invention, their pharmaceutically acceptable salts, solvates, tautomers, or stereoisomers, or pharmaceutically acceptable solvates of any of the above, or pharmaceutical compositions of any of the above, can be administered systemically and / or topically to specific organs.

[0279] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may be administered as a single dose or as a chronic dose. Chronic administration means that the methods and compositions of the present invention are administered multiple times to a given individual. For example, as will be apparent to those skilled in the art, chronic administration may involve administering multiple doses of the pharmaceutical composition to an animal, including an individual, once daily, twice daily, or more or less frequently. In other embodiments, the methods and compositions are administered acutely. Acute administration means that the methods and compositions of the present invention are administered at a time close to or concurrent with an ischemic or obstructive event. This means that, for example, acute administration may involve administering one or more doses of a pharmaceutical composition at the time of an ischemic or obstructive event such as acute myocardial infarction, for example, in the early stages of an ischemic or obstructive event such as stroke, or before, during, or after a surgical procedure. The timing close to or contemporaneous with the ischemic or obstructive event will vary depending on the ischemic event, but could be, for example, within about 30 minutes after the onset of symptoms of myocardial infarction, stroke, or intermittent claudication. In certain embodiments, acute administration is administration within about 1 hour of the ischemic event. In certain embodiments, acute administration is administration within about 2 hours, 6 hours, 10 hours, 12 hours, 15 hours, or 24 hours after the ischemic event.

[0280] In certain embodiments, the compounds of the present invention or their pharmaceutical compositions can be administered chronically. In certain embodiments, chronic administration may include the periodic administration of multiple intravenous injections within a day. In certain embodiments, chronic administration may include the administration of a single intravenous injection as a bolus or as a continuous infusion daily, approximately every other day, approximately every 3 to 15 days, approximately every 5 to 10 days, and in certain embodiments, approximately every 10 days.

[0281] Combination therapy In certain embodiments, the compounds of the present invention, their pharmaceutically acceptable salts, solvates, tautomers, or stereoisomers, or pharmaceutically acceptable solvates of any of the above, may be used in combination therapy with at least one other therapeutic agent. The compounds of the present invention and other therapeutic agents may act additively or, in certain embodiments, synergistically. Depending on the embodiment, the compounds of the present invention may be administered concurrently with other therapeutic agents, such as compounds for treating diseases associated with dysfunction of energy metabolism; compounds for treating muscle fatigue; compounds for improving muscle strength and endurance; compounds for enhancing the viability of transplanted organs; and compounds for improving the viability of isolated cells. Depending on the embodiment, the compounds of the present invention, pharmaceutically acceptable salts, or pharmaceutically acceptable solvates of any of the above may be administered before or after the administration of other therapeutic agents, such as compounds for treating diseases associated with energy metabolic dysfunction such as ischemia and ventricular hypertrophy, neurodegenerative diseases such as ALS, Huntington's disease, Parkinson's disease, or Alzheimer's disease, surgery-related ischemic tissue injury, and reperfusion tissue injury; compounds for treating multiple sclerosis (MS); compounds for treating mental disorders such as schizophrenia, bipolar disorder, or anxiety; compounds for treating muscle fatigue; compounds for improving muscle strength and endurance; compounds for enhancing the viability of transplanted organs; and compounds for improving the viability of isolated cells.

[0282] The pharmaceutical composition of the present invention may include, in addition to one or more compounds of the present invention, one or more therapeutic agents effective in treating the same or different diseases, disorders, or symptoms.

[0283] The method of the present invention involves the administration of one or more compounds or pharmaceutical compositions of the present invention and one or more other therapeutic agents, provided that the combination of administrations does not inhibit the therapeutic effect of one or more compounds of the present invention, and / or cause adverse effects from the combination.

[0284] In certain embodiments, the composition of the present invention can be administered concurrently with the administration of another therapeutic agent, which may be part of a pharmaceutical composition or dosage form containing the compound of the present invention, or may be contained in a composition or dosage form separate from the one containing the compound of the present invention. In certain embodiments, the compound of the present invention can be administered before or after the administration of another therapeutic agent. In certain embodiments of combination therapy, Therapy may involve, for example, altering the dosage between a composition of the present invention and a composition containing another therapeutic agent to minimize adverse side effects associated with a specific drug. When a compound of the present invention is administered concurrently with another therapeutic agent that has the potential to cause adverse side effects, not limited to toxicity, the therapeutic agent can be conveniently administered at a dose below the threshold at which adverse side effects are induced.

[0285] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may be administered to a patient with or in combination with other compounds for treating Parkinson's disease, such as amantadine, benztropine, bromocriptine, levodopa, pergolide, pramipexole, ropinirole, selegiline, trihexyphenidyl, or any combination thereof.

[0286] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may be administered to a patient with or in combination with other compounds for treating Alzheimer's disease, such as donepezil, galantamine, memantine, rivastigmine, tacrine, or any combination thereof.

[0287] In certain embodiments, the compound or pharmaceutical composition of the present invention may be administered to a patient together with, or with, another compound for treating ALS, such as riluzole.

[0288] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may be administered to a patient with or in combination with other compounds for the treatment of ischemic stroke, such as aspirin, nimodipine, clopidogrel, pravastatin, unfractionated heparin, eptifbatide, beta-blockers, angiotensin-converting enzyme (ACE) inhibitors, enoxaparin, or any combination thereof.

[0289] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may include, or be administered to a patient together with, other compounds for treating ischemic cardiomyopathy or ischemic heart disease, such as ACE inhibitors, e.g., ramipril, captopril, and lisinopril; n-blockers, e.g., acebutolol, atenolol, betaxolol, bisoprolol, carteolol, nadolol, pembutolol, propranolol, timolol, metoprolol, carvedilol, and aldosterone; diuretics; digitoxin, or any combination thereof.

[0290] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may be administered to a patient with or in combination with other compounds for treating cardiovascular diseases, such as antithrombotic agents, cholesterol-lowering agents, antiplatelet agents, vasodilators, beta-blockers, angiotensin blockers, digitalis and its derivatives, or any combination thereof.

[0291] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may be administered to a patient together with, or contain, another compound for the treatment of MS. Examples of drugs useful for treating MS include corticosteroids, e.g., methylprednisolone; IFN-β, e.g., IFN-β1a and IFN-β1b; glatiramer acetate (Copaxone®); monoclonal antibodies that bind to late-stage antigen 4 (VLA-4) integrin (Tysabri®), e.g., natalizumab; immunomodulators, e.g., FTY720 sphingosine-1-phosphate modifiers; and COX-2 inhibitors, e.g., BW755c, piroxicam, and phenidone; as well as glutamate excitotoxicity and iNOS inhibitors, free radical scavengers, and cation channel blockers. Neuroprotective therapies including memantine; AMPA antagonists, such as topiramate; and glycine-binding site NMDA antagonists are also mentioned (Virley, NeruoRx 2005, 2(4), 638-649, and citations herein; and U.S. Patent Application No. 2004 / 0102525).

[0292] In certain embodiments, the compound or pharmaceutical composition of the present invention may be administered to a patient together with, or containing, another compound for the treatment of schizophrenia. Examples of antipsychotics useful for treating schizophrenia include, but are not limited to, acetophenazine, artheroxylone, amitriptyline, aripiprazole, astemizole, benzquinamide, carphenazine, chlormezanone, chlorpromazine, chlorprothixen, clozapine, desipramine, droperidol, haloperidol, fluphenazine, flupentixol, glycine, roxapine, mesolidazine, morindone, olanzapine, ondansetron, perphenazine, pimozide, prochlorperazine, procyclidine, promazine, propiomazine, quetiapine, remoxiprid, reserpine, risperidone, certindol, sulpiride, terfenadine, thiethylperazine, thioridazine, thiothixen, trifloperazine, triflupromazine, trimeprazine, and ziprasidone. Other antipsychotics useful for treating the symptoms of schizophrenia include amisulpride, paraperidone, blonanserin, butaperazine, carfenadine, eprivanserin, iloperidone, lamictal, osanetant, paliperidone, perospirone, piperacetadine, lacloprid, remoxiprid, salizotan, sonepiprazole, sulpiride, ziprasidone, and zotepine; serotonin and dopamine (5HT / D2) agonists, such as asenapine and bifepurnox; neurokinin 3 antagonists, such as tarnetant and osanetant; amphakines, such as CX-516, galantamine, memantine, modafinil, ocaperidone, and tolcapone; and alpha-amino acids, such as D-serine, D-alanine, D-cycloserine, and N-methylglycine.

[0293] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may be administered to a patient with or in combination with other compounds for treating bipolar disorder, such as aripiprazole, carbamazepine, clonazepam, clonidine, lamotrigine, quetiapine, verapamil, and ziprasidone.

[0294] In certain embodiments, the compounds or pharmaceutical compositions of the present invention may be administered to a patient with or in combination with other compounds for treating anxiety, such as alprazolam, atenolol, buspirone, chlordiazepoxide, clonidine, chlorazepic acid, diazepam, doxepin, escitalopram, harazepam, hydroxyzine, lorazepam, prochlorperazine, nadolol, oxazepam, paroxetine, prochlorperazine, trifloperazine, and venlafaxine. [Examples]

[0295] The following examples illustrate in detail assays for characterizing the compounds of the present invention and the use of the compounds of the present invention. As will be obvious to those skilled in the art, numerous modifications can be made to both the materials and methods without departing from the scope of this disclosure.

[0296] General Experiments The NMR spectrum of the compound is obtained at 400 or 500 MHz ( 1 Measured at 25°C (H). 1 Unless otherwise specified, 1H NMR spectra were processed with a 0.3 Hz linewidth broadening. LC / MS analysis was performed using Shimadzu LCMS 2010 (column: sepax ODS 50×2.0 mm, 5 μm), Agilent 1200 HPLC, and 1956 MSD (column: Waters XBridge C18 4.6×50 mm). A 3.5mm) Shim-pack XR-ODS (30×3.0, 2.2um) was used in ES(+) ionization mode with an Agilent 3110TM (or Agilent Zorbax Bonus RP®, 2.1×50mm, 3.5μm). Exemplary settings were a temperature of 50°C and a flow rate of 0.8 mL / min, an injection volume of 2 μL, mobile phase A = water with 0.1% formic acid and 1% acetonitrile added, mobile phase B = methanol with 0.1% formic acid added; retention time was in minutes. Method details: (i) Binary Pump G1312B® with UV / Vis diode array detector G1315C and Agilent The HPLC was operated with a 6130™ mass spectrometer, in positive and negative ion electrospray mode, with UV detection at 220 and 254 nm. Mobile phase B was increased from 5% to 95% with a linear gradient of 2.5 mins (II), maintained at 95% B for 0.5 mins (III), and decreased from 95% to 5% with a linear gradient of 0.1 mins (IV), maintained at 5% B for 0.29 mins. For analytical HPLC sample analysis, an Agilent 1200 Series™ HPLC was used with a Waters HSS T3™ column, 2.1 × 50 mm, 1.8 μm, at a temperature of 60°C and a flow rate of 0.5 mL / min. Mobile phase A = water with 0.1% formic acid and 0.1% acetonitrile added, mobile phase B = acetonitrile with 0.1% formic acid added; retention times were in minutes. Melting points were recorded using a Thomas Hoover Unimelt™ capillary melting point analyzer. The reaction progress was recorded using a Merck Thin-layer chromatography on EMD 60 F254 silica gel glass plates was monitored by visualization using UV light and / or iodine treatment. Chromatographic purification was performed using a Teledyne ISCO CombiFlash Companion® at various flow rates from 5 to 100 mL / min. The columns used were Teledyne ISCO RediSep disposable flash columns (pre-packed with 4, 12, 24, 40, 80, or 120 g of silica gel). Peaks were detected by variable wavelength UV absorption (200–360 nm).Preparative reverse-phase chromatography was performed using a Gilson 215 Liquid Handler equipped with a Varian Model 218 pump and operated with Chromeleon® software. Detection was performed using either a Varian Pro Star® UV-Vis or a Sedex55® ELSD unit. Chromatographic separation was performed using a Phenomenex Kinetex® 5u C18 100A, Axia, 100×30 mm column at a flow rate of 28 mL / min.

[0297] Compounds tested by bioassay, such as compounds A, B, C, D, E, F, G, H, J, K, L, and M, correspond to compounds exemplified in the synthesis procedures described herein. For example, compound E is the compound of Example 26, step 5A, as described in this application.

[0298] Example 1: Method for confirming enzymatic cleavage of prodrugs in vitro For creatine prodrugs, it is generally desirable that the prodrug remains unchanged (i.e., uncleaved) while in systemic circulation and is cleaved in target tissue (i.e., releasing the parent drug). A useful level of stability can be determined, at least in part, by the mechanism and pharmacokinetics of the prodrug. A useful level of instability can also be determined, at least in part, by the pharmacokinetics of the prodrug and the parent drug (e.g., creatine) in systemic circulation and / or the gastrointestinal tract when administered orally. Generally, prodrugs that are more stable in the gastrointestinal tract (assessed by stability in artificial gastric fluid, artificial intestinal fluid, intestinal S9, pancreatin, or colon lavage assays) and more unstable in mouse plasma, rat plasma, human plasma, mouse, rat, and / or human liver S9, liver microsomes, and / or hepatocyte specimens may be useful as orally administered prodrugs. Generally, mouse plasma, rat plasma, human plasma, mouse, rat, and / or human liver S9, liver microsomes, and / or hepatocyte specimens are more stable, as are target tissue cell lysates or isolated target tissue cell specimens, such as brain, muscle, and Caco-2 S9 specimens. Therefore, more unstable prodrugs may be useful as systemically administered prodrugs and / or may be more effective in delivering prodrugs to target tissues. Generally, prodrugs that are more stable in physiological buffers at various pH levels may be more useful as prodrugs. Generally, prodrugs that are more unstable in target tissue cell lysates and / or isolated target tissue cell specimens, such as brain, muscle, and Caco-2 S9 specimens, may be cleaved intracellularly and release the parent drug to the target tissue. Results of tests that reveal enzymatic or chemical cleavage of prodrugs in vitro, such as those described in this example, can be used to select prodrugs for in vivo testing.

[0299] The stability of prodrugs can be evaluated in one or more in vitro systems using various specimens and methods known in the field. Tissues and specimens should be obtained from the distributor (e.g., Pel-Freez Biologicals, Rogers, Ark., or GenTest Corporation, Woburn, Mass.). Useful experimental conditions for in vitro testing are listed in Table 1. The prodrug should be added to each specimen in triplicate.

[0300] [Table 1]

[0301] Samples containing alkaline phosphatase are tested in and out of the presence of a phosphatase inhibitor cocktail (Sigma). Samples are incubated at 37°C for a period ranging from 30 minutes to 24 hours. At each time point, the sample is quenched with 50% ethanol. The baseline concentration of the prodrug is determined by directly adding the compound to the 50% ethanol / sample mixture (t=0). The sample is centrifuged at 14,000 rpm for 15 minutes, and the concentrations of the unchanged prodrug and released parent drug are measured using LC / MS / MS. This stability of the prodrug against specific enzymes (e.g., peptidase) is also assessed in vitro by incubation with the purified enzyme.

[0302] The pancreatin stability test is performed by incubating a prodrug (5 μM) with 1% (w / v) pancreatin (Sigma, P-1625, derived from porcine pancreas) in 0.025 M Tris buffer (pH 7.5) containing 0.5 M NaCl at 37°C. The reaction is stopped by adding 50% ethanol by 3 times the volume. After centrifugation at 14,000 rpm for 15 minutes, the supernatant is removed and analyzed for prodrug, creatine, and creatinine by LC / MS / MS.

[0303] To determine the stability of the prodrug in artificial gastric juice (SGF), a prodrug (10 μM) was incubated in SGF (0.2% NaCl w / v, 0.7% HCl v / v, pH 1.2) at 37°C, with and without pepsin (purified pepsin with 800-2500 units of activity per 1 mg of protein, 3.2 g per liter). At selected time points (e.g., 0, 15, 30, 60, and 120 minutes), 50 μL was taken, neutralized with 50 μL of 0.1 M sodium bicarbonate solution, and 150 μL of ice-cold acetonitrile was added. The samples were centrifuged at 4°C at 4,000 × g for 15 minutes, and the supernatant was collected and analyzed for prodrug, creatine, and creatinine concentrations by LC-MSMS (Table 2).

[0304] [Table 2]

[0305] To ensure stability in artificial intestinal fluid (SIF), a prodrug (10 μM) was used in SI. Incubate the samples at 37°C in F (0.68% KH2PO4 w / v, 0.86% NaOH v / v, pH 6.8) with and without pancreatin (1% w / v). At selected time points (e.g., 0, 15, 30, 60, and 120 minutes), take 50 μL samples and add 150 μL of ice-cold acetonitrile to terminate the incubation. Centrifuge the samples at 4°C at 4,000 × g for 15 minutes, and collect the supernatant. Analyze the prodrug, creatine, and creatinine concentrations by LC-MSMS (Table 3).

[0306] [Table 3]

[0307] To determine the stability of Caco-2 cells in Caco-2 lysate S9, Caco-2 cells are grown for 21 days before being harvested. The culture medium is removed, the cell monolayer is rinsed and scraped off, and placed in 10 mM sodium phosphate / 0.15 M potassium chloride, pH 7.4, chilled on ice. The cells are sonicated at 4°C using a probe sonicator to lyse them. The lysed cells are then transferred to a 1.5 mL centrifuge vial and centrifuged at 9,000 g for 20 minutes at 4°C. The resulting supernatant (Caco-2 cell lysate S9 fraction) is divided equally into 0.5 mL vials and stored at -80°C until use.

[0308] For stability testing, the prodrug (5 μM) is incubated in the S9 fraction of Caco-2 cell lysates (0.5 mg / mL 0.1 M Tris buffer, pH 7.4) at 37°C. Three sets of samples are quenched at each time point using 50% ethanol. The initial (t=0) concentration of the prodrug is determined by directly adding 5 μM of the prodrug to the 50% ethanol / Caco-2 cell lysate mixture. The samples are subjected to LC / MS / MS analysis to determine the concentrations of the prodrug, creatine, and creatinine.

[0309] To determine the prodrug stability in the plasma of mice, rats, humans, or other species, the prodrug (10 μM) or positive control (10 μM, propantheline or procaine) is incubated in undiluted plasma. Two sets of samples, one prodrug and one control, are prepared and analyzed. The stock prodrug solution is prepared in DMSO (10 mM) and diluted to 0.1 mM in pH 7.4 phosphate buffer to prepare a spike solution. A fixed volume (10 μL) of the prodrug spike solution is placed in a 96-well plate. Warm (37°C) plasma is used. Add (90 μL) to the wells planned for time points 5, 15, 30, 45, and 60 minutes; at t=0 min, add the quench solution (400 μL acetonitrile) directly to the prodrug-containing well, followed by 90 μL of warmed plasma. At 5, 15, 30, 45, and 60 minutes, add 400 μL of acetonitrile to the well to stop the reaction. After quenching, shake the plate for 10 minutes (600 rpm), then centrifuge at 5500 g for 15 minutes. Transfer a fixed volume (50 μL) to an analysis plate and dilute with 100 μL of ultrapure water (Millipore) for LC-MSMS quantification of the prodrug concentration, and optionally creatine and / or creatinine. Select a chromatography column (e.g., Atlantis HILIC Silica, Gemini C-18, Ultimate XB-C18) based on the lipophilicity and polarity of each prodrug. The LC-MSMS platform (e.g., Sciex API4000, Sciex API6500) is also selected based on the requirements of each prodrug. The stability of the compounds (prodrugs) disclosed herein in mouse plasma incubation is shown in Table 4, and the stability in human plasma incubation is shown in Table 5.

[0310] [Table 4]

[0311] [Table 5]

[0312] For the liver microsome stability test, the prodrug or positive control (testosterone, propafenone, diclofenac, 7-ethoxycoumarin, or propranolol) is incubated (in pairs) at 5 μM in liver or intestinal fractions derived from mouse, human, dog, monkey, and / or rat. Incubation is performed at 37°C, with or without an NADPH regeneration system, to indicate whether metabolism proceeds via NADPH-dependent enzymes (i.e., P450, FMO, NADPH-P450 reductase, or other oxidase enzymes). Pairs of the prodrug and positive control are prepared and analyzed. The stock prodrug is prepared in DMSO (10 mM) and diluted to 0.05 mM in a 25% MeOH / pH 7.4 phosphate buffer mixture to prepare a spike solution. A fixed volume (10 μL) of the prodrug spike solution is placed in a 96-well plate. Add 80 μL of warmed (37°C) microsome solution to the wells planned for time points 5, 15, 30, 45, and 60 minutes, incubate for 10 minutes, and then initiate the reaction with 10 μL of NADPH regeneration solution. For t=0 minutes, add 300 μL of quench solution (acetonitrile) directly to the prodrug-containing well, followed by the microsome solution and NADPH solution. Incubate the prodrug in a thermally inactivated fraction or buffer to distinguish between enzymatic and non-enzymatic degradation. At the specified time points (e.g., 0, 5, 10, 20, 30, and 60 minutes), collect samples and stop them with an equal volume of cold acetonitrile containing an appropriate internal standard (e.g., labetalol, tolbutamide). After quenching, centrifuge the plate at 4000 g for 20 minutes. A fixed amount (100 μL) is transferred to an analysis plate and diluted with 400 μL of ultrapure water (Millipore) for quantification of the prodrug concentration and, if applicable, creatine and / or creatinine using LC-MSMS. Based on the lipophilicity and polarity of the drug, a chromatography column (e.g., ACE 5) Select Phenyl (Phenomenex C18 Synergi Hydro-RP, Atlantis HILIC Silica). The LC-MSMS platform (e.g., Sciex API4000, Sciex API6500) is also selected based on the requirements of each prodrug. The metabolic stability of the compounds disclosed herein in mouse liver microsome incubation is shown in Table 6, and the metabolic stability in human liver microsome incubation is shown in Table 7.

[0313] [Table 6]

[0314] [Table 7]

[0315] For S9 stability testing, the prodrug (5 μM) is incubated at 37°C in S9 lysates from the liver or intestines of mouse, human, dog, monkey, and / or rat (0.5 mg / mL 0.1 M potassium phosphate buffer, pH 7.4, 1 mM NADPH). Incubation is performed in or out of the presence of an NADPH regeneration system to indicate whether metabolism proceeds via NADPH-dependent enzymes (i.e., P450, FMO, NADPH-P450 reductase, or other oxidase enzymes). The triple prodrug is quenched with 50% ethanol at each time point. The initial (t=0) concentration of the prodrug is determined by directly adding 5 μM of the prodrug to the 50% ethanol / S9 lysate mixture. The samples are subjected to LC / MS / MS analysis to determine the concentrations of the prodrug, creatine, and creatinine.

[0316] For hepatocyte stability testing, the prodrug (5 μM) is incubated with seeded hepatocytes (e.g., mouse, rat, or human). Fresh hepatocytes seeded in 12-well plates with overlays (except for rats, which do not have overlays) are received (LifeTechnologies). Upon receipt, the transport medium is immediately removed and replaced with 1 mL of warmed culture medium. The cells are climate-acclimatized overnight at 37°C in a 5% CO2 atmosphere. The medium is aspirated from the plate and the prodrug (5 μM) or solvent control (0. Replace with 1 mL of fresh medium containing 0.125% DMSO. Incubate the samples (three sets) at 37°C in a 5% CO2 atmosphere for 0, 0.25, 0.5, 0.75, 1, 2, and 4 hours. Incubate an additional well containing a solvent control for calibration curve preparation and background measurement. Remove the medium and freeze at the selected time. Wash the cells twice with cold PBS. Add 0.5 mL of cold 70% acetonitrile containing an internal standard to each well and gently scrape the cells from the plate. Aspirate the recovered cells suspended in the organic solution into a vial and freeze at -80°C. For analysis, remove the cell saturation in 70% ACN from the freezer, disrupt it, and vortex. Add 500 μL of water to each test tube and vortex the sample again. Centrifuge the test tubes at 4°C at 13000 rpm for 10 minutes. The cell supernatant and the original recovered culture medium are extracted and analyzed by LC-MS / MS to determine the prodrug, creatine, and creatinine.

[0317] The chemical stability of the prodrug was determined using three buffer solutions: (1) 0.1 M potassium phosphate, 0.5 M NaCl, pH 2.0; (2) 0.1 M Tris-HCl, 0.5 M NaCl, pH 7.4; and (3) 0.1 M Tris-HCl, 0.5 M NaCl, pH 8.0. The prodrug (5 μM) was added to each buffer solution in a triple chain. The sample was quenched with 50% ethanol at each time point. The initial (t=0) concentration of the prodrug was determined by directly adding 5 μM of the prodrug to a 50% ethanol / pH buffer mixture. The sample was subjected to LC / MS / MS analysis to determine the concentrations of the prodrug, creatine, and creatinine.

[0318] Example 2: In vitro measurement of creatine release from prodrugs To assess the creatine-releasing ability of prodrugs and their ability to preferentially redirect creatine to undesirable cyclization to creatinine, d3-labeled (deuterium-labeled methyl group) prodrugs are incubated with liver lysates (e.g., mouse, human) specially prepared to preserve N-reductase activity. The use of d3-labeled prodrugs is essential for distinguishing prodrug-derived creatine (d3-creatine) from high concentrations of endogenous (unlabeled) creatine. Incubation (37°C) is performed in 100 mM potassium phosphate buffer, pH 6.0 to optimize N-reductase activity. Prodrugs are tested at final concentrations of 20 μM and 200 μM. Approximately 4-5 mg of lysate is used in each reaction. A cofactor (NADH) is included at a final concentration of 1 mM. Benzamido oxime (final concentration 500 μM) is used as a positive control for N-reductase activity. N-reductase activity is confirmed by the conversion of benzamido oxime to benzamidine. Negative controls include incubation without NADH (to assess NADH-independent prodrug cleavage) and incubation without liver lysates (with NADH) to assess non-enzymatic prodrug cleavage under assay conditions. Prodrug incubation is prepared by adding 10 μL of prodrug stock solution (400 or 4000 μM, 40% DMSO-containing water) to 100 μL of potassium phosphate buffer, followed by 70 μL of liver lysates. The reaction is initiated by adding 20 μL of NADH solution (10 mM) or 20 μL of water for the (-)NADH negative control. At selected points (e.g., 0, 30, 60, and 180 minutes), a 50 μL sample is taken and the reaction is stopped by adding 150 μL of ice-cold acetonitrile (80% ACN / 20% water) stop solution. The sample was centrifuged at 15890 × g for 10 minutes at 4°C, and the supernatant was then transferred to storage at 40°C until LC-MSMS analysis was determined. The sample supernatant was analyzed by LC-MSMS (HILIC column) to determine the levels of d3-prodrug, d3-creatine, and d3-creatinine (Table 8).

[0319] [Table 8]

[0320] Example 3: In vitro measurement of Caco-2 cell permeability of prodrugs The passive permeability of creatine prodrugs is assessed in vitro using standard methods well known in the field (see, e.g., Stewart, et al., Pharm. Res., 1995, 12, 693). For example, passive permeability can be evaluated by testing the flux of the prodrug across a cultured polarized cell monolayer (e.g., Caco-2 cells).

[0321] Caco-2 cells obtained from continuous culture (passage number less than 28) were seeded at high density on a Transwell polycarbonate filter. The cells were maintained until the day of the experiment in DMEM / 10% fetal bovine serum + 0.1 mM non-essential amino acids + 2 mM L-Gln, 5% CO2 / 95% O2, 37°C. Permeability tests were performed at apical pH 6.5 (in 50 mM MES buffer containing 1 mM CaCl2, 1 mM MgCl2, 150 mM NaCl, 3 mM KCl, 1 mM NaH2PO4, and 5 mM glucose) and basolateral pH 7.4 (in Hanks equilibrium salt solution containing 10 mM HEPES) in the presence of efflux pump inhibitors (250 μM MK-571, 250 μM verapamil, 1 mM ofloxacin). Perform the following steps: Place the insert in a 12-well or 24-well plate containing buffer and incubate at 37°C for 30 minutes. Add the prodrug (100 μM, 250 μM, 300 μM, or 500 μM) to the apical or basolateral compartment (donor) and determine the concentrations of the prodrug and / or released parent drug (creatine) in the opposite compartment (receiver) at 1-hour intervals using LC / MS / MS. Determine the apparent permeability (P app The value of ) is calculated using the following formula: P app =V r (dC / dt) / (AC o ) In the formula, Vr is the volume (mL) of the receiver compartment; dC / dt is the total flux (μM / s) of the prodrug and parent drug, determined from the slope of the concentration-time graph in the receiver compartment; C o A is the initial concentration of the prodrug (μM); and A is the surface area of ​​the membrane (cm²). 2 In a particular embodiment, a prodrug with remarkable transcellular permeability is ≥1 × 10 -6 P in cm / s app This indicates a value, and in a particular embodiment, ≥ 1 × 10 -5 P in cm / s app This indicates a value, and in a particular embodiment, ≥ 5 × 10 -5 P in cm / s app This shows the value.

[0322] Example 4: Uptake by Caco-2 cells and HEK-2 cells Seed Caco-2 or HEK peaks at 250,000 and 500,000 cells / well in polylysine-coated 24-well plastic cell culture plates, respectively. Incubate cells overnight at 37°C. Add the prodrug to each well in 1 mL of fresh medium. Test each concentration of the prodrug in triplicates. Add only medium to the control well. Wash the cells four times with Hanks equilibrium salt solution at each time point. Extract the compounds by lysing the cells and adding 200 μL of 50% ethanol to each well and letting stand at room temperature for 20 minutes. Take a fixed volume of the ethanol solution and transfer it to a 96-well V-bottom plate, and centrifuge at 5,700 rpm for 20 minutes at 4°C. Analyze the supernatant by LC / MS / MS to determine the concentrations of the prodrug, creatine, and / or creatinine.

[0323] Example 5: SMVT expression in mammalian cells The sodium-dependent multivitamin transporter (SMVT; product of the SLC5A6 gene) was subcloned into a plasmid (TREX plasmid, Invitrogen Inc., Carlsbad Calif.) that enables tetracycline-induced expression. The SMVT expression plasmid was transfused into human embryonic kidney (HEK) cell lines, and stable clones were isolated by G418 sorting and fluid-labeled cell sorting (FACS). Biotin uptake in SMVT-HEK cell clones was validated. SMVT-HEK / TREX cells were seeded at 100,000 cells / well in 96-well plates, incubated at 37°C for 24 hours, and then tetracycline (1 μg / mL) was added to each well for another 24 hours to induce SMVT transporter expression. Radiolabeling was performed. 3 H-biotin (approximately 100,000 cpm / well) was added to each well. The plate was incubated at room temperature for 10 minutes. 3 H-biotin was removed, and the cells were washed three times in a 96-well plate washer with cold assay solution. Scintillation solution was added to each well, the plate was sealed, and the cells were counted using a 96-well plate compatible scintillation counter.

[0324] Using a similar method, HEK cells expressing other transporters or SMVT or other cell lines expressing other transporters can be prepared.

[0325] The GenBank acceptance number for human SMVT is NM.021095, which is incorporated herein by reference. References relating to the SMVT transporter include the amino acid sequence described or encoded by GenBank reference number NM.021095, as well as alleles, homogenes, and derivative variants and fragments of that sequence that retain essentially the same transporter activity. Typically, such variants exhibit at least 90% sequence identity with exemplary GenBank nucleic acids or amino acid sequences. The substrates of SMVT are free carboxylic acids and Short alkyl chains whose terminals are cyclic or branched groups, for example, C 1-6These are alkyl-containing compounds. Examples of SMVT substrates include biotin, pantothenic acid, and 4-phenylbutyric acid.

[0326] Example 6: Competitive assay using SMVT A competitive binding assay was developed to determine whether creatine prodrugs bind to the SMVT transporter. This assay measures the extent to which the uptake of radiolabeled substrates, such as biotin or pantothenic acid, is blocked when the concentration of the test compound is varied. The maximum half-inhibitory concentration (IC) of the inhibition of substrate transport by the test compound is measured. 50 ) is an indicator of the affinity of the test compound to the SMVT transporter. If the test compound competes with the radiolabeled substrate to bind to SMVT, less radiolabeled substrate will be transported to HEK cells. For test compounds that do not interact with SMVT in a manner that competes with the substrate, the curve remains essentially flat, i.e., no dose-response is observed. The amount of radiolabeled substrate taken up by cells is measured by lysing the cells and measuring the radioactivity number per minute. The competitive binding test is performed as follows: SMVT-HEK / TREX cells are seeded at 100,000 cells / well in a 96-well plate, left at 37°C for 24 hours, then tetracycline (1 μg / mL) is added to each well and left for another 24 hours to induce SMVT transporter expression. Radiolabeled cells are then subjected to various concentrations of unlabeled biotin or pantothenic acid in 2- or 3-row systems, both in the presence and absence of these concentrations. 3 Add H-biotin (approximately 100,000 cpm / well) to each well. Incubate the plate at room temperature for 10 minutes. 3 Remove H-biotin and wash the cells three times with cold assay solution in a 96-well plate washer. Add scintillation solution to each well, seal the plate, and count using a 96-well plate compatible scintillation counter. Graph the data using Prism software (GraphPad, Inc., San Diego, Calif.) and analyze it using nonlinear regression analysis.

[0327] Example 7: Treatment of HEK SMVT cells with creatine prodrug The uptake of unlabeled creatine prodrugs is measured in HEK cells that stably express SMVT. Cells are seeded at a density of 250,000 cells / well in a polylysine-coated 24-well tissue culture plate. After 24 hours, cells are treated with tetracycline (1 μg / mL) to induce SMVT expression, or left untreated. The assay is performed the following day (approximately 48 hours after seeding). Creatine prodrugs (final concentration 0.1 mM) are added to buffered saline (HBSS), and 0.5 mL of each test solution is added to each well. Cells are allowed to take up the test compound for 1 or 3 hours. The test solution is aspirated, and the cells are washed four times with ice-cold HBSS. The cells are then lysed in 50% ethanol solution (0.2 mL / well) at room temperature for 15 minutes. The lysate is centrifuged at 5477 × g for 15 minutes at 4°C to remove cell fragments. The concentrations of creatine prodrugs and creatine in cells are determined by analytical LC / MS / MS. Transporter-specific uptake is determined by comparison with control cells lacking transporter expression.

[0328] Example 8: Effect of treatment on the creatine kinase system HEK cells expressing SMVT are treated with buffer, creatine prodrug (100 μM), creatine (100 μM), or creatine analog (100 μM) for the specified time according to the protocol of Example 6. After treatment, intracellular concentrations of creatine prodrug, creatine phosphate, ATP, and creatine and / or creatine analog are measured by analytical LC / MS / MS.

[0329] Example 9: Restoration of cellular energy homeostasis after sodium azide treatment Applying the method described by Weinstock and Shoham, Neural Transm. 2004, 111(3), 347-66, intracellular energy The protective effect of the present invention's compounds on ghee homeostasis is evaluated.

[0330] HEK TREX SMVT cell lines are seeded in 24-well polylysine-coated tissue culture plates at 250k per well. The following day, cells are treated with doxycycline (1 μg / mL) to express the SMVT transporter. This transporter is necessary for the effective uptake of the creatine prodrug under test, e.g., the compound of the present invention. Cells are incubated and assayed the following day. Cells are washed twice with glucose-free HBSS buffer. Cells are then incubated at 37°C in a 5% CO2 incubator in the same buffer, with or without sodium azide, at 20 mM. The typical range of sodium azide used in these experiments is 1 mM to 9 mM. After this, the creatine analog prodrug is added to the cells at 300 μM, or the cells are left untreated. In some experiments, creatine is used as a comparison. Cells are incubated for a further 20 minutes and then washed with buffer. The sample was extracted with 50% ethanol for 15 minutes and processed for LC / MS / MS to detect creatine prodrug, creatine, and ATP levels. The increase in creatine phosphate and ATP levels after contact with the creatine prodrug in sodium azide-treated cells indicates that the prodrug can restore cellular energy homeostasis.

[0331] Example 10: Protection against 3-nitropropionic acid-induced toxicity The protective effect of the compounds of the present invention on intracellular energy homeostasis will be evaluated by applying the method described by Brouillet et al., J. Neurochem 2005, 95(6), 1521-40.

[0332] The rat cardiomyocyte cell line H9c2 was obtained from ATCC (#CRL-1446). A 20 mM stock solution of 3-nitropropionic acid (3-NP) was prepared immediately before use in standard medium (DMEM / high glucose (4.5 g / L) / 10% FBS / 6 mM L-glutamine / PSF), and 1N sodium hydroxide was added dropwise to adjust the pH to 7.4. A 40 mM stock solution of a creatine prodrug, such as the compound of the present invention, was prepared in DMSO, and creatine was directly dissolved in serum-free medium at a concentration of 10 mM.

[0333] To measure the degree of cytoprotection against 3-NP toxicity provided by creatine prodrugs and / or creatine analogs, H9c2 cells are seeded at 10K cells per well in standard medium in a 96-well clear-bottom black tissue culture plate and incubated overnight at 37°C. The following day, the medium is removed and replaced with serum-free medium containing creatine prodrugs or a series of creatine dilutions. The plate is incubated at 37°C for 2 hours. The medium is then aspirated and removed, replaced with standard medium containing various concentrations of 3-NP, and the plate is incubated at 37°C for a further 20 hours. To determine the number of viable cells in each well, an equal volume of CellTiter-Glo reagent (Promega) is added at room temperature and mixed in a plate shaker for 10 minutes. The luminescence is measured by reading the plate with a luminometer. The luminescence produced in this assay is proportional to the amount of ATP present and is directly related to the number of metabolically active cells.

[0334] If the viability of cells in contact with 3-NP and creatine prodrugs increases compared to the viability of cells in contact with 3-NP and creatine, it indicates that creatine prodrugs have the ability to maintain cellular energy homeostasis.

[0335] Example 11: Pharmacokinetics of creatine prodrug after colonic administration in rats Sustained-release oral dosage forms, which slowly release the drug over a period of approximately 6 to 24 hours, generally release a significant proportion of the dose in the colon. That is, drugs suitable for use in such dosage forms are those that should be absorbed in the colon. This experiment involved the compounds of the present invention, including creatine. This procedure assesses the uptake of the creatine prodrug and creatine in biological fluids such as plasma / blood or cerebrospinal fluid (CSF) after intracolonic administration of the prodrug, and the resulting levels, in order to determine the suitability of the creatine prodrug for use in a sustained-release oral dosage form. The bioavailability of the creatine prodrug and creatine after co-administration of the creatine prodrug can be calculated for oral and / or intracolonic administration of the creatine prodrug.

[0336] Process A: Dosage Protocol Purchase rats and pre-treat them with cannulas in both the ascending colon and jugular vein. The animals should be conscious during the experiment. All animals should be fasted overnight and for 4 hours after administration of the creatine prodrug. Administer the creatine prodrug directly into the colon via the cannula as a solution (dissolved in water or other suitable solvent or vehicle) at doses equivalent to approximately 1 mg to 200 mg per kg of body weight. Obtain blood samples (0.3 mL) from the jugular vein cannula at 8-hour intervals and immediately quench them with sodium metabisulfite or other suitable antioxidant to prevent oxidation of the creatine prodrug. The blood samples may be further quenched with methanol / perchloric acid to prevent hydrolysis of the creatine prodrug after sampling. Analyze the blood samples as described below. Samples may also be collected from CSF or other suitable biological fluids.

[0337] Step B: Sample preparation for prodrugs absorbed from the colon Add methanol / perchloric acid (300 μL) to an empty 1.5 mL Eppendorf test tube. Collect rat blood (300 μL) at various time points and place them in EDTA test tubes containing 75 μL of sodium metabisulfite, then vortex and mix. Immediately place a fixed amount of blood (100 μL) into an Eppendorf test tube and vortex and mix. Add 10 microliters of standard stock solutions of creatine prodrugs (0.04, 0.2, 1, 5, 25, and 100 μg / mL) and 10 μL of 10% sodium metabisulfite solution to 80 μL of empty rat blood to prepare the final calibration standards (0.004, 0.02, 0.1, 0.5, 2.5, and 10 μg / mL). Then add methanol / perchloric acid (300 μL in a 50 / 50 ratio) to each test tube, followed by 20 μL of p-chlorophenylalanine. Vortex the sample and centrifuge at 14,000 rpm for 10 minutes. Analyze the supernatant by LC / MS / MS.

[0338] Step C: LC / MS / MS analysis An API 4000 LC / MS / MS spectrometer, equipped with an Agilent 1100 binary pump, a CTC HTS-PAL autosampler, and a Zorbax XDB C8 4.6×150mm column, is used during analysis. Appropriate mobile phases, such as (A) 0.1% formic acid and (B) acetonitrile containing 0.1% formic acid, can be used. Appropriate gradient conditions, such as 5%B for 0.5 minutes, then 98%B for 3 minutes, maintaining 98%B for 2.5 minutes, then returning to 2%B for 2 minutes, can be used. A TurboIonSpray ion source is used with the API 4000. Analysis is performed in the appropriate ion mode, and the MRM transition for each analyte is optimized using standard solutions. 5 μL of each sample is injected. Non-compartmental analysis is performed on individual animal characteristics using WinNonlin software (v.3.1 Professional version, Pharsight Corporation, Mountain View, Calif.). A statistical summary of the major parameter estimates is C max (Peak concentration observed after drug administration), T max (The point at which the maximum concentration is reached is the point when the peak concentration is observed), AUC (0-t)(Area under the serum concentration-time curve from time 0 to the final blood collection, estimated using the logarithmic trapezoidal method), AUC (0-。infin。) (The area under the blood concentration-time curve from time 0 to infinity, estimated by extrapolating to infinity using the log-linear trapezoidal method until the final blood collection), and t 1 / 2 This is done for z (final half-life).

[0339] The pharmacokinetic parameters of the creatine prodrug and creatine after colonic administration of the creatine prodrug are determined and compared with those obtained after equivalent colonic administration of the creatine prodrug. The maximum blood concentration (C) of the creatine prodrug and creatine is determined. max If the blood concentration-time area (AUC) after colonic administration of the creatine prodrug is higher than that obtained with colonic administration of the creatine prodrug, it indicates that the prodrug improves colon bioavailability.

[0340] Example 12: Pharmacokinetics of creatine prodrugs after intravenous or oral administration to rats Each group consists of 4-6 mature male Sprague-Dawley rats (approximately 250g) and is administered creatine prodrug either as an intravenous bolus injection or via oral gastric tube feeding. The animals are conscious during the experiment. When administered orally, the creatine prodrug is administered as an aqueous solution (or as a solution in another suitable solvent, optionally containing a suitable vehicle) at an appropriate dose equivalent per kg of body weight. Blood samples (0.3 mL) are obtained via jugular vein cannula at intervals of 8 hours after oral administration. The blood is immediately quenched, for example, with acetonitrile containing 1% formic acid, and then frozen at ±80°C until analysis. Samples can also be taken from CSF or other suitable biological fluids.

[0341] Add 300 (300) μL of acetonitrile containing 0.1% formic acid to an empty 1.5 mL test tube. Collect 300 μL of rat blood at various time points, place them in test tubes containing EDTA, and vortex to mix. Immediately add a fixed amount of blood (100 μL) to a test tube and vortex to mix. Add 10 microliters of creatine prodrug standard stock solutions (0.04, 0.2, 1, 5, 25, and 100 μg / mL) to 90 μL of empty rat blood quenched with 300 μL of acetonitrile containing 0.1% formic acid. Then, add 20 μL of p-chlorophenylalanine to each test tube to prepare the final calibration standards (0.004, 0.02, 0.1, 0.5, 2.5, and 10 μg / mL). Vortex the samples and centrifuge at 14,000 rpm for 10 minutes. The supernatant is analyzed by LC / MS / MS.

[0342] An API 4000 LC / MS / MS spectrometer, equipped with an Agilent 1100 binary pump, a CTC HTS-PAL autosampler, and a Phenomenex Synergihydro-RP 4.6×30mm column, is used for analysis. Appropriate mobile phase and gradient conditions are used for the analysis. The analysis is performed in the appropriate ion mode, and the MRM transition for each analyte is optimized using standard solutions. 5(5) μL of each sample is injected. Non-compartmental analysis is performed on individual animal characteristics using WinNonlin (v.3.1 professional version, Pharsight Corporation, Mountain view, Calif.). Statistical summaries of key parameter estimates are obtained using C max (Peak concentration observed after drug administration), T max (The point at which the maximum concentration is reached is the point when the peak concentration is observed), AUC (0-t) (Area under the serum concentration-time curve from time 0 to the final blood collection, estimated using the logarithmic trapezoidal method), AUC (0-。infin。) (The area under the blood concentration-time curve from time 0 to infinity, estimated by extrapolating to infinity using the log-linear trapezoidal method until the final blood collection), and t 1 / 2 This will be done regarding the (final half-life).

[0343] The oral bioavailability (F(%)) of creatine prodrugs is determined by comparing the area under the creatine prodrug concentration-time curve (AUC) after oral administration with the AUC of the creatine prodrug concentration-time curve after intravenous administration, based on a normalized dose.

[0344] The pharmacokinetics of creatine prodrugs and creatine can also be determined by obtaining samples from CSF. If the levels of creatine prodrugs and / or creatine are high, it can be said that the prodrug has the ability to cross the blood-brain barrier.

[0345] Similar studies on the pharmacokinetics of creatine prodrugs can be conducted in other animals, including, but not limited to, dogs, monkeys, and humans.

[0346] Example 13: Use of an animal model to assess the efficacy of creatine prodrugs for the treatment of amyotrophic lateral sclerosis (ALS). A mouse model of SOD1 mutation-associated ALS has been developed. In this model, mice express a human superoxide dismutase (SOD) mutant glycine-alanine at residue 93 (SOD1). These SOD1 mice exhibit dominant acquisition of undesirable traits of SOD, and develop motor neuron degeneration and dysfunction similar to human ALS (Gurney et al., Science 1994, 264(5166), 1772-1775; Gurney et al., Ann. Neurol. 1996, 39, 147-157; Gurney, J. Neurol. Sci. 1997, 152, S67-73; Ripps et al., Proc Natl Acad Sci USA 1995, 92(3), 689-693; and Bruijn et al., Proc Natl Acad Sci USA 1997, 94(14), 7606-7611). SOD1 gene-transformed mice show signs of hindlimb weakness at approximately 3 months of age and die at 4 months of age. Common features with human ALS include increased astrocytes, microgliosis, oxidative stress, elevated cyclooxygenase / prostaglandin levels, and, as a result of disease progression, severe motor neuron loss.

[0347] The study will be conducted in transgenic mice (B6SJL-TgN(SOD1-G93A) 1 Gur) overexpressing the human Cu / Zn-SOD G93A mutation, non-transgenic B6 / SJL mice, and their wild-type littermates. Mice will be housed in a 12-hour diurnal / light cycle (starting at 45 days of age) and allowed to freely approach either a solid sample supplemented with the test compound or a standard cold-pressed solid sample molded into the same pellet as a control. Genotyping can be performed at 21 days of age as described in Gurney et al., Science 1994, 264(5166), 1772-1775. SOD1 mice will be divided into several groups and treated with the test compound or used as controls.

[0348] Mice are observed daily and weighed weekly. To assess their health, mice are weighed weekly and changes in lacrimation / salivation, eyelid closure, ear spasms and pupillary responses, whisker direction, postural and righting reflexes, and overall health scores are examined. Systemic pathological examination is performed at the time of sacrifice.

[0349] The motor coordination ability of an animal can be assessed by one or more methods known to those skilled in the art. For example, motor coordination can be assessed using a neurological scoring method. In neurological scoring, the neurological score of each limb is observed and recorded according to a defined 4-point scale: 0 = normal hind limb reflex (the animal extends the hind limb when lifted by the tail); 1 = abnormal hind limb reflex (the animal does not extend the hind limb when lifted by the tail); 2 = abnormal limb reflex and evidence of paralysis; 3 = absence of reflex and complete paralysis; and 4 = inability to return to the correct posture within 30 seconds when placed on its side, or confirmed death. The primary endpoint is survival, and the secondary endpoints are neurological score and body weight. Neurological score observation and body weight are performed and recorded 5 days a week. Data analysis is performed using appropriate statistical methods.

[0350] The rotorod test assesses an animal's ability to remain on a rotating rod, enabling evaluation of motor coordination and proprioceptive sensitivity. The apparatus is an automatically rotating rod with a diameter of 3 cm. For example, a rod that rotates 12 times per minute. The rotor rod test measures how long a mouse can maintain its balance on the rod without falling. This test can be stopped after any time limit, for example, 120 seconds. If an animal falls before 120 seconds, its ability is recorded and the test is repeated twice. The average time over the three tests is calculated. Motor defects are indicated by a decrease in walking time.

[0351] In the grid test, mice are placed on a grid (length: 37 cm, width: 10.5 cm, mesh opening: 1 × 1 cm) located on a flat support. 2 The mouse is placed on a grid. The number of times the mouse touches its feet through the grid is counted and used as a measure of motor coordination.

[0352] The hanging test evaluates an animal's ability to cling to a wire. The apparatus consists of a wire stretched horizontally 40 cm above a platform. The animal is instructed to grasp the wire with its front legs. The test is performed three times in a row, and the time it takes the animal to grasp the wire with its hind legs is recorded (maximum 60 seconds).

[0353] Electrophysiological measurements (EMG) can also be used to assess motor activity. Electromuscular recordings are taken using an EMG recording device. Mice are anesthetized during EMG observation. The parameters measured are the amplitude and latency of the compound muscle action potential (CMAP). CMAP is measured in the gastrocnemius muscle after sciatic nerve stimulation. A reference electrode is inserted near the Achilles tendon, and a probe needle is placed on the tail ridge. A grounding needle is inserted into the lower back of the mouse. The sciatic nerve is stimulated with a single 0.2 msec pulse at hypermaximal intensity (12.9 mA). Amplitude (mV) and response latency (ms) are measured. Amplitude indicates the number of active motor units, and distal latency reflects the motor nerve conduction velocity.

[0354] The efficacy of the test compound can also be evaluated using biomarker analysis. To assess the regulation of protein biomarkers in SOD1 mice with motor dysfunction, lumbar spinal cord samples (protein extracts) are added to protein chip arrays with various surface chemical / biochemical properties and analyzed, for example, by surface-enhanced laser desorption / ionization time-of-flight mass spectrometry. Subsequently, the protein expression characteristics of various treatment groups are compared from the data using an integrated protein mass characterization method. The analysis can be performed using appropriate statistical methods.

[0355] Example 14: Clinical trial to assess the efficacy of creatine prodrugs for the treatment of Parkinson's disease The efficacy of creatine prodrugs in the treatment of Parkinson's disease can be assessed using the following clinical trials. Patients with idiopathic Parkinson's disease who meet the Queen Square Brain Bank criteria (Gibb et al., J Neurol Neurosurg Psychiatry 1988, 51, 745-752) and have motor variability and a defined short-term GABA analog response (1.5–4 hours) are eligible to participate. A further requirement is that the patient has clinically relevant peak-dose dyskinesia after taking their medication each morning in their current regimen. Patients are also required to be stable on a fixed-dose regimen for at least one month prior to the start of the clinical trial. Patients whose current medication regimen includes sustained-release L-Dopa, COMT inhibitors, selegiline, anticholinergics, or other medications that may interfere with gastric absorption (e.g., antacids) will be excluded. Other exclusion criteria include patients with psychotic symptoms or those undergoing treatment for psychotropic illness, patients with clinically relevant cognitive impairment, and patients with a score of less than 24 on the MMS (Short Scale for Calculation of Intelligence) (Folstein et al., J Psychiatr Res 1975, 12, 189-198) Other conditions include potential pregnancy, off-state Hoehn & Yahr stage 5, severe unstable diabetes mellitus, and unstable cardiovascular disease or moderate to severe renal or hepatic dysfunction. Complete blood count, liver function, and renal function blood tests will be performed at baseline and after completion of the study.

[0356] A randomized, double-blind, crossover study design will be used. The pharmacokinetics of creatine prodrugs and released creatine can be assessed by measuring blood concentrations at appropriate time intervals. Creatine levels in the brain can also be measured non-invasively by magnetic resonance spectroscopy (MRS).

[0357] For clinical evaluation, motor function is assessed using the UPDRS (Unified Parkinson's Disease Rating Scale) motor score and the Brain test (Giovanni et al., J Neurol Neurosurg Psychiatry 1999, 67, 624-629). The Brain test involves the patient typing on a laptop keyboard with the weaker hand. These tests are performed at baseline, then immediately after each blood draw until the patient reaches their full "on" state, and then at intervals until the patient reaches their baseline "off" state. Once the patient reaches their full "on" state, video recordings are taken three times at 20-minute intervals. The following memory and motor tasks have been shown to increase dyskinesia (Duriff et al., Mov Disord 1999, 14, 242-245), so observe these tasks during each video session: (1) sitting still for one minute; (2) doing mental arithmetic; (3) putting on a coat and buttoning it; (4) pouring water into a glass and drinking it; and (5) walking. Record the potential increase in dyskinesia induced by the test compound by scoring the videotapes using, for example, multiple versions of the Goetz Rating Scale and the Abnormal Involuntary Movements Scale.

[0358] The actual incidence and severity of dyskinesia are monitored by a dyskinesia monitor (Manson et al.). The measurement is performed using the device (al., J Neurol Neurosurg Psychiatry 2000, 68, 196-201). This device is taped to the shoulder on the side of the patient's condition that is worsening. The monitor records throughout the entire task session, providing a measure of the frequency and severity of dyskinesia occurrences.

[0359] The results can be analyzed using appropriate statistical methods.

[0360] Example 15: Efficacy of creatine prodrug in an animal model of MPTP-induced neurotoxicity in Parkinson's disease MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) is a neurotoxin that causes Parkinson's disease-like syndrome in both humans and experimental animals. Research into the mechanism of MPTP neurotoxicity is focused on its major metabolite, MPTP. + This indicates that the generation of MPP is involved in this toxicity. + MPTP is formed by the action of monoamine oxidase on MPTP. Monoamine oxidase inhibitors block the neurotoxicity of MPTP in both mice and primates. + The neurotoxic effect is specific to dopaminergic neurons because of MPP mediated by synaptic dopamine transporters. + It appears to be due to the uptake of this transporter. The blocking agent for this transporter is MPP + Prevent neurotoxicity. MPP + MPTP has been shown to be a relatively specific inhibitor of mitochondrial complex I activity, binding to complex I at the rotenone binding site and impairing oxidative phosphorylation. In vivo studies have shown that MPTP may deplete striatal ATP concentration in mice. MPTP administered intrastriatically in rats. + It has been demonstrated that this leads to a significant depletion of ATP, as well as an increase in lactate concentration, specifically in the striatum at the injection site. Compounds that enhance ATP production may offer protection against MPTP toxicity in mice.

[0361] Administer creatine prodrugs to animals such as mice or rats for 3 weeks, then perform MPTP treatment. Administer MPTP for 1 week at an appropriate dose, interval, and mode of administration. The animals are then slaughtered. The control group is given either normal saline or MPTP hydrochloride. After slaughter, the two striae are rapidly dissected and placed in cold 0.1 M perchloric acid. Subsequently, the tissue is sonicated, and a certain amount is analyzed for protein content using a fluorescence spectrometer assay. Dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) are also quantified. The concentrations of dopamine and its metabolites are expressed in nmol / mg protein.

[0362] Creatine prodrugs that offer protection against MPTP-induced DOPAC depletion, HVA, and / or dopamine depletion are neuroprotective and therefore may be useful in the treatment of Parkinson's disease.

[0363] Example 16: Evaluation of potential anti-Parkinson's disease activity using a haloperidol-induced spontaneous motor impairment animal model. Adenosine antagonists such as theophylline have been shown to reverse the behavioral inhibitory effects of dopamine antagonists such as haloperidol in rodents, and are being investigated as a valid method for screening drugs with potential anti-Parkinson's disease activity (Mandhane, et al., Eur. J. Pharmacol. 1997, 328, 135-141). The ability of creatine prodrugs to block haloperidol-induced deficiency of spontaneous motor activity in mice can be used to assess the in vivo activity and potential of anti-Parkinson's disease activity.

[0364] Mice used in the experiment are acclimatized to the climate by being housed in a controlled environment before being used in the experiment. 0.2 mg / kg of haloperidol is administered to the mice 1.5 hours before the test. This dose reduces spontaneous motility by at least 50% from baseline. The test compound is administered 5–60 minutes before the test. The animals are then individually placed in clean, transparent polycarbonate cages with flat, perforated lids. The cages are placed in a frame equipped with a 3x6 array of photocells, which are linked to a computer used to tally light blocking, to measure horizontal spontaneous motility. The mice are allowed to move freely for one hour, and the number of times light is blocked during this period is used as an indicator of spontaneous motility. This is compared to data from control animals to determine if there is a statistically significant difference.

[0365] Example 17: Animal model of 6-hydroxydopamine in Parkinson's disease The neurochemical deficits observed in Parkinson's disease can be reproduced by topically injecting the dopaminergic neurotoxin 6-hydroxydopamine (6-OHDA) into brain regions containing either the cell bodies or axonal fibers of nigrostriatal neurons. By unilaterally damaging the nigrostriatal pathway on only one side of the brain, behavioral asymmetry in motor impairment can be observed. Although animals with unilateral damage remain motorable and self-sufficient, the remaining dopamine-sensitive neurons on the damaged side become hypersensitive to stimuli. This is demonstrated by the observation that animals exhibit a clear rotation in the direction opposite to the damaged side after systemic administration of dopamine agonists such as apomorphine. The ability of 6-OHDA-damaged rats to induce counter-rotation has been shown to be a sensitivity model that predicts drug efficacy in the treatment of Parkinson's disease.

[0366] Male Sprague-Dawley rats are housed in a controlled environment and acclimatized before being used in the experiment. Fifteen minutes before the procedure, the animals are intraperitoneally injected with the noradrenergic reuptake inhibitor desipramine (25 mg / kg) to prevent damage to non-dopaminergic neurons. The animals are then placed in an anesthesia chamber and anesthetized with a mixture of oxygen and isoflurane. Once unconscious, the animals are moved to a stereotactic frame and anesthesia is maintained through a mask. The top of the animals' heads are shaved and sterilized with iodine solution. Once dry, a 2 cm long incision is made along the midline of the scalp, the skin is pushed back and secured with clips to expose the skull. Next, a small hole is drilled into the skull above the injection site. With the aim of damaging the substantia nigra striatal pathway, the injection cannula is slowly lowered over the right medial forebrain bundle, 3.2 mm anterior-posteriorly from the cruciate suture, 1.5 mm medial-laterally, and 7.2 mm from the dura mater. Two minutes after lowering the cannula, 6-OHDA is injected at a rate of 0.5 μL / min for more than four minutes to a final dose of 8 μg. The cannula is left in place for another five minutes to allow diffusion to occur, and then slowly withdrawn. The skin is then sutured closed, and the animal is removed from the stereotactic frame and returned to its enclosure. The rats are allowed to recover from the surgery for two weeks, after which behavioral tests are performed.

[0367] Rotational behavior is measured using a rotometer system. This system consists of a stainless steel bowl (45 cm in diameter x 15 cm in height) enclosed by a transparent plexiglass cover that extends 29 cm in height around the rim of the bowl. To assess rotation, the rat is placed over a cloth jacket fitted with a tethering spring connected to an optical rotometer positioned above the bowl. The optical rotometer assesses movement to the left or right, whether partial (45°) or complete (360°) rotation.

[0368] To reduce stress during administration of the test compound, rats are initially accustomed to the device for 15 minutes for four consecutive days. On the test day, the rats are given the test compound, e.g., creatine prodrug. Immediately before the test, the animals are given a subthreshold dose of apomorphine by subcutaneous injection, then fitted with a harness and the number of rotations is recorded for one hour. The total number of complete counter-rotations during the one-hour test period is used as an indicator of anti-Parkinson's disease efficacy.

[0369] Example 18: Animal experiment to assess the efficacy of creatine prodrugs in ischemic injury. Mature male rats were administered creatine prodrug, and approximately 24 hours later, they were anesthetized and prepared for coronary artery occlusion. An additional dose of creatine prodrug was administered at the start of the procedure to occlude the left main coronary artery for 30 minutes, after which it was released. Subsequently, the same dose of creatine prodrug was administered postoperatively at appropriate intervals and durations. The animals were then examined for cardiac function. Animals given a placebo injection (saline) showed a significant increase in left end-diastolic pressure and diastolic rigidity of the heart secondary to myocardial infarction. Creatine prodrug, which eliminates or reduces the deficit in cardiac function compared to placebo-treated controls, is useful in preventing ischemic injury.

[0370] Example 19: Animal experiment to assess the ability of creatine prodrugs to maintain organ viability. Wistar male rats weighing 300-330g were administered a creatine prodrug or vehicle, and 24 hours later, their hearts were removed for ex vivo testing. The animals were sacrificed using pentobarbital (0.3 mL) and intravenous heparinization (0.2 mL). The hearts were first allowed to equilibrate for 15 minutes. Then, the left ventricular balloon was inflated to a volume that produced an end-diastolic pressure of approximately 8 mmHg. A left ventricular pressure-volume curve was constructed by inflating the balloon volume in increments of 0.02 mL. Zero volume was defined as the point when the left ventricular end-diastolic pressure was zero. At the end of the pressure-volume curve, the left ventricular balloon was deflated to return the end-diastolic pressure to 8 mmHg, and after confirming coronary blood flow, a resting period of 15 minutes was maintained. The hearts were then stopped with 50 mL of Celsior+ molecules and allowed to rest under 60 cmH2O pressure at 4°C. Next, the heart is removed, filled with the same solution, and placed in a plastic container surrounded by crushed ice, and stored at 4°C for 5 hours.

[0371] After storage, the heart is transferred to a Langendorff apparatus. The balloon catheter is reinserted into the left ventricle and reinflated to the same volume as during the pre-ischemic period. The heart is reperfused at 37°C for at least 2 hours. The reperfusion pressure is set to 50 cmH2O for the first 15 minutes of reperfusion, and then reduced to 100 cmH2O for the next 2 hours. Pacing (320 beats per minute) is resumed. Isovolumetric measurements of systolic index and diastolic pressure are performed in a triple series at 25, 45, 60, and 120 minutes of reperfusion. At this point, a pressure-volume curve is obtained, and coronary effluent during the 45-minute reperfusion is collected to assess creatine kinase leakage. The leakage was measured. Improvements in left ventricular pressure, as well as improvements in the volumetric pressure curve, a decrease in diastolic left ventricular pressure, and a reduction in creatine kinase leakage after treatment with a creatine analog prodrug demonstrate the ability of creatine prodrugs to maintain organ viability.

[0372] Example 20: Neuroprotective effect of creatine analog prodrug in a gene-transformed mouse model of Huntington's disease N171-82Q genetically modified HD mice and non-genetically modified littermates were treated with a creatine analog prodrug or vehicle from 10 weeks of age. The mice were placed on a rotating rod ("rotor rod"). The length of time until the mouse fell off the rotor rod was recorded as a measure of motor coordination. The total distance the mouse moved was also recorded as a measure of total spontaneous movement. Mice administered with the neuroprotective creatine prodrug in the N171-82Q genetically modified HD mouse model stayed on the rotor rod longer and traveled longer distances than mice administered with the vehicle.

[0373] Example 21: Efficacy of creatine prodrug in a malonic acid model of Huntington's disease A range of reversible and irreversible inhibitors of enzymes involved in energy production pathways have been used to create animal models of neurodegenerative diseases such as Parkinson's disease and Huntington's disease. Inhibitors of succinate dehydrogenase, an enzyme that affects cellular energy homeostasis, have been used to create a model of Huntington's disease (Brouillet). et al., J. Neurochem. 1993, 60, 356-359; Beal et al., J. Neurosci. 1993, 13, 4181-4192; Henshaw et al., Brain Research 1994, 647, 161-166 (1994); and Beal et al., (J. Neurochem. 1993, 61, 1147-1150). The succinate dehydrogenase enzyme plays a central role in both the tricarboxylic acid cycle and the mitochondrial electron transport chain. Malonic acid is a reversible inhibitor of succinate dehydrogenase. Intrastriatal injection of malonic acid in rats has been shown to induce dose-dependent striatal excitotoxic lesions that are attenuated by both competitive and non-competitive NMDA antagonists (Henshaw et al., Brain Research 1994, 647, 161-166). Lamotrigine, a glutamate-releasing inhibitor, also attenuates the lesions. Co-injection with succinate blocks the lesions, consistent with its effect on succinate dehydrogenase. The lesions are accompanied by a significant decrease in ATP levels and a significant increase in lactate levels in vivo, as shown by chemical shift resonance imaging (Beal et al., J. Neurochem. 1993, 61, 1147-1150). The lesions result in the same pattern of cell saving seen in Huntington's disease, suggesting that malonate loading is a useful model for the neuropathological and neurochemical features of Huntington's disease.

[0374] To evaluate the effect of creatine prodrugs in this malonic acid model for Huntington's disease, male Sprague-Dawley rats are administered creatine prodrugs at appropriate doses, intervals, and routes. The prodrug is administered for two weeks, followed by malonic acid administration, and the rats are sacrificed one week later. Malonic acid is dissolved in distilled deionized water and the pH is adjusted to 7.4 with 0.1 M HCl. A 1.5 μL intrastriatal injection containing 3 μmol of malonic acid is administered to the left striatum at the level of the cruciate suture, 2.4 mm lateral to the midline and 4.5 mm ventral to the dura mater. The animals are decapitated and sacrificed on day 7, and the brains are quickly removed and placed in ice-cold 0.9% physiological saline. The brains are sectioned at 2 mm intervals along the brain shape. The sections are then placed posteriorly downwards in 2% 2,3,5-triphenyltetrazolium chloride. The sections are stained in the dark at room temperature for 30 minutes, then removed and placed in 4% paraformaldehyde pH 7.3. The lesions, which have been lightly stained and become visible, are placed on the posterior surface of each section. The evaluation is performed across the surface. The measurements are validated by comparing them with measurements obtained from adjacent Nissl-stained sections.

[0375] Compounds that exert neuroprotective effects and are therefore useful in the treatment of Huntington's disease show a reduction in malonate-induced lesions.

[0376] Example 22: Efficacy of creatine prodrug in a model of creatine transporter dysfunction A mouse model of human CrT deficiency has been created, making it possible to develop treatments for this condition (Skelton et al., PloS One, 201, 6(1), e16187). Mice possessing exons 2-4 of Slc6a8 adjacent to the loxP site were crossed with Cre:CMV mice to create a line of ubiquitous CrT knockout-expressing mice. Male CrT- / y (affected) mice lack Cr in the brain and muscles, and Cr is significantly reduced in other tissues, including the heart and testes. CrT- / y mice show increased pathway length during acquisition and reverse learning in the Morris water maze. During probe testing, CrT- / y mice show increased mean distance from platform location. Compared to CrT+ / y mice, CrT- / y mice show decreased novel object recognition and conditioned fear memory. CrT- / y mice have increased serotonin and 5-hydroxyindoleacetic acid in the hippocampus and prefrontal cortex. Ubiquitous CrT knockout mice exhibit learning and memory deficits similar to those in human CrT deficiency; therefore, this model is useful for understanding this disorder and testing creatine prodrugs as a treatment for it.

[0377] To evaluate the effects of creatine prodrugs in the Morris Water Maze (MWM), male CrT- / y mice will be administered creatine prodrugs at appropriate doses, intervals, and routes. The MWM is a test of spatial learning and reference memory (Vorhees et al., (Nature Protocols 2006, 1:848-858) The animals are tested as described in Skelton et al., Brain Res 2003, 984:1-10 and Schaefer et al., Neuroscience 2009, 164:1431-1443. Before testing on the hidden platform, the animals are trained (cued learning) on ​​a visible platform for 6 days. During this stage, the maze is surrounded by curtains to block out any visible distant cues, and a brass rod with an orange ball attached is placed on a 10 cm diameter platform, which is then placed in a predetermined quadrant. On day 1, the animals are given 6 trials (90 seconds each) using this platform from the same starting position; from the following day onward, the test is given twice a day, with the starting position and platform position randomly determined.

[0378] The hidden platform portion of the MWM trial was conducted in three phases (6 days / phase: acquisition, reversal, and transition). Each phase consisted of 6 days of testing four times per day for the animals to learn the hidden platform, followed by a single-probe test (without platform) on day 7 (Vorhees et al., Nature Protocols 2006, 1:848-858). The platform diameters were 10 cm for acquisition, 7 cm for reversal (placed in opposing quadrants), and 5 cm for transition (placed in one of adjacent quadrants). Ability was measured using AnyMaze software (Stoelting Company, Wood Dale, IL). The effect of prodrug treatment was analyzed by comparing performance between control (untreated male CrT- / y mice and / or wild-type mice) and prodrug-treated mice.

[0379] To evaluate the effects of creatine prodrugs in a conditioned fear model, male CrT- / y mice were administered creatine prodrugs at appropriate doses, intervals, and routes. Cues and contextual fear were assessed as described in Peters et al., Science 2010, 328: 1288-1290). Day 1, untreated. (Control) and treated (prodrug-administered) mice are exposed to a set of 30 buzzers (82 dB, 2 kHz, 30-second on / off cycles), followed by 3 buzzers and electric shocks to the feet (0.5 mA for 1 second). The following day, as a test of contextual fear, the animals are returned to a chamber without buzzers or electric shocks. The next day, the animals are placed in a new grate-bed chamber. After 3 minutes of climate acclimatization, the buzzer is sounded and freezing behavior is scored. The animals are then exposed to 30 cycles of 30 seconds of buzzer sound followed by 30 seconds of silence, and the duration of fear is measured. Freezeframe software and Coulbourn test chambers are used (Coulbourn Instruments, Allentown, PA). Percentage of freezing time is analyzed. The effect of prodrug treatment is analyzed by comparing performance between control (untreated male CrT- / y mice and / or wild-type mice) and prodrug-treated mice.

[0380] To evaluate the effect of creatine prodrugs in a novel object recognition (NOR) model, male CrT- / y mice were administered creatine prodrugs at appropriate doses, intervals, and routes. NOR is a test of short-term memory (Clark et al., J Neurosci 2000, 20: 8853-8860). Mice were acclimatized to the test area (91 cm in diameter) for two days (10 minutes / day), followed by two days (10 minutes / day) of acclimatization to two identical objects. On the test day, the animals were presented with two novel identical objects until the cumulative observation time reached 30 seconds. After one hour, the animals were tested for memory by being presented with a copy of one of the familiar objects and the novel object. The discrimination index was calculated by subtracting the observation time of the familiar object from the time spent observing the novel object. The effects of prodrug treatment will be analyzed by comparing the results of prodrug-treated mice with those of control mice (untreated male CrT- / y mice and / or wild-type mice).

[0381] Compounds useful for treating creatine transporter disorders can be evaluated using one or more of the assessment methods outlined above, or alternative models testing behavior, neurological function, and / or neuromuscular function, comparing treated male CrT levels to untreated controls. - / y The improvement will be shown in mice.

[0382] Example 23: Synthesis of ethyl (N'-hydroxy-N-methylcarbamimidoamide) acetate JPEG2026048846000104.jpg29123 Ethyl (N'-hydroxy-N-methylcarbamimidoamide) acetate can be synthesized using the procedure described in Zbinden et al., Bioorganic & Medicinal Chemistry Letters, 2005, 15: 5344-5352. Briefly, ethyl [cyano(methyl)amino] acetate (available from MP Biomedicals, Inc.) is refluxed in EtOH with hydroxylamine hydrochloride to obtain the title compound.

[0383] Example 24: Synthesis of tert-butyl=2-(3-(tert-butoxycarbonyl)-2-hydroxy-1-methylguanidino)acetate-3-yl)amino]acetate JPEG2026048846000105.jpg36132 Step 1: Synthesis of tert-butyl=2-(N-methylcyanamide)acetate JPEG2026048846000106.jpg12128 In a round-bottom flask equipped with a stirring bar, tert-butyl=2-(methylamino)acetate (500 mg, 2.75 mmol), potassium carbonate (761 mg, 5.50 mmol), and acetonitrile (10 mL) were added. The mixture was stirred at room temperature for 30 minutes, and then a solution of cyanogen bromide (320 mg, 3.025 mmol) in acetonitrile (2 mL) was added. The reaction mixture was stirred at room temperature overnight, and the solvent was removed by decantation, leaving an insoluble residue. The solvent was then evaporated under reduced pressure, and the product was purified by flash chromatography using petroleum ether-ethyl acetate (gradient from 0% to 40% ethyl acetate) to obtain tert-butyl=2-(N-methylcyanamide)acetate (410 mg, 2.41 mmol, yield 88%) as a white solid. ES LC-MS m / z = 171 (M+H + ) & 193 (M+Na + ).

[0384] Step 2: Synthesis of tert-butyl=2-(2-hydroxy-1-methylguanidino)acetate JPEG2026048846000107.jpg17128 In a round-bottom flask equipped with a stirring bar, tert-butyl=2-(N-methylcyanamide) acetate (300 mg, 1.76 mmol) and tetrahydrofuran (5 mL) were added. To this mixture, hydroxylamine (50% aqueous solution, 583 mg, 8.80 mmol) was added. After 1 hour, 10 mL of water was added, and the mixture was extracted three times with 5 mL of dichloromethane. The organic layer was then dried over MgSO4, filtered, and concentrated under reduced pressure to obtain crude tert-butyl=2-(2-hydroxy-1-methylguanidino) acetate (349 mg, 1.72 mmol, 98% yield) as a white solid. This was immediately used in the next step. ES LC-MS m / z = 204 (M+H + ).

[0385] Step 3: Synthesis of tert-butyl=2-(3-(tert-butoxycarbonyl)-2-hydroxy-1-methylguanidino)acetate-3-yl)amino]acetate JPEG2026048846000108.jpg18128 In a round-bottom flask equipped with a stirring bar, tert-butyl=2-(2-hydroxy-1-methylguanidino)acetate (349 mg, 1.72 mmol) and tetrahydrofuran (5 mL) were added. Di-tert-butyl bicarbonate (375 mg, 1.72 mmol) was added to this mixture. The mixture was stirred overnight at room temperature. The solvent was then evaporated under reduced pressure, and the product was purified by flash chromatography using dichloromethane-ethyl acetate (gradient from 0% to 30% ethyl acetate) to obtain tert-butyl=2-(3-(tert-butoxycarbonyl)-2-hydroxy-1-methylguanidino)acetate (151 mg, 0.50 mmol, yield 29%) as a white solid. ES LC-MS m / z = 304 (M+H + ). 1 1H NMR (dimethyl NMR) Rufoxide-d) δ: 5.73 (s, 2H), 3.81 (s, 2H), 2.76 (s, 3H), 1.42 (s, 9H), 1.41 (s, 9H).

[0386] Example 25: Synthesis of tert-butyl=2-(3-(tert-butoxycarbonyl)-2-hydroxy-1-triduteriomethylguanidino)acetate JPEG2026048846000109.jpg66133 Step 1: Synthesis of tert-butyl=2-(4-nitrophenylsulfonamide) acetate JPEG2026048846000110.jpg16128 Under nitrogen, 20 g, 119.76 mmol, tert-butyl ester hydrochloride glycine and 260 mL, pyridine were added to a round-bottom flask equipped with a stirring bar. The mixture was cooled to 0°C, and then 28.98 g, 131.74 mmol, 4-nitrobenzenesulfonyl chloride was added little by little while maintaining the temperature of the mixture below 10°C. The reaction mixture was then raised to room temperature. After 18 hours at room temperature, the reaction mixture was poured into 1000 mL of water. The precipitate was filtered and vacuum-dried to obtain tert-butyl=2-(4-nitrophenylsulfonamide) acetate (30.8 g, 97.46 mmol, yield 81%) as a yellow solid. 1 H NMR(CDCl3)δ: 8.36-8.34 (m, 2 H), 8.07-8.05 (m, 2H), 5.23 (brs, 1H), 3.75 -3.74 (d, J = 5.6 Hz, 2H),1.35 (s, 9H).

[0387] Step 2: Synthesis of tert-butyl=2-[(4-nitrophenyl)sulfonyl-(triduteriomethyl)amino]acetate JPEG2026048846000111.jpg16128 Under nitrogen, tert-butyl=2-(4-nitrophenylsulfonamide)acetate (30.8 g, 97.46 mmol), DMF (320 mL), and CD3I (14.13 g, 97.46 mmol) were added to a round-bottom flask equipped with a stirring bar. To this mixture, Cs2CO3 (34.85 g, 107.22 mmol) was added at room temperature, and the reaction mixture was stirred for 45 minutes. The reaction mixture was then poured into water (1000 mL) and extracted with SiO2 (3 × 500 mL). The organic phases were combined, washed with NaCl (500 mL), dried, filtered, and the solvent was evaporated under reduced pressure to obtain tert-butyl=2-[(4-nitrophenyl)sulfonyl-(triduteriomethyl)amino]acetate (27.8 g, 83.48 mmol, yield 81%) as a pale yellow solid. 1 1H NMR (CDCl3) δ: 8.36-8.33 (d, J = 8.8 Hz,2H), 8. 01-7.99 (d, J = 9.2 Hz, 2H), 3.98 (s, 2H), 1.38 (s, 9H).

[0388] Step 3: Synthesis of tert-butyl=2-(tert-butoxycarbonyl(triduteriomethyl)amino)acetate JPEG2026048846000112.jpg15128 Under nitrogen, tert-butyl=2-[(4-nitrophenyl)sulfonyl-(triduteriomethyl)amino]acetate (27.8 g, 83.48 mmol), Cs2CO3 (67.83 g, 208.7 mmol), acetonitrile (400 mL), and THF (40 mL) were added to a round-bottom flask equipped with a stirring bar. Thiophenol (34 mL, 333.93 mmol) was added to this solution, and the reaction mixture was heated at 45°C for 90 minutes. The reaction mixture was then diluted with MTBE (500 mL) and extracted with water (5 × 100 mL). The water extracts were combined and washed with MTBE (500 mL), and DCM (500 mL), followed by (BOC)2O (36.4 g, 166.97 mmol), was added to the water mixture. The two-phase reaction mixture was vigorously stirred overnight. Next, the phases were separated, and the aqueous layer was extracted with DCM (500 mL x 5). The organic phases were combined and dried (Na2SO4), filtered, and the solvent was evaporated under reduced pressure. The product was purified by chromatography using a 120 g silica cartridge with heptane-siRNA (gradient of siRNA from 0% to 30%) to obtain tert-butyl=2-(tert-butoxycarbonyl(triduteriomethyl)amino)acetate (6 g, 24.19 mmol, yield 28%) as a colorless oil. 1 H NMR(CDCl3)δ: 3.84-3.74 (m, 2H), 1 0.45-1.41 (m, 18H).

[0389] Step 4: Synthesis of tert-butyl=2-(N-triduteriomethylamino)acetate TFA salt JPEG2026048846000113.jpg15128 In a round-bottom flask equipped with a stirring bar, tert-butyl=2-(tert-butoxycarbonyl(triduteriomethyl)amino)acetate (500 mg, 2.02 mmol) and dichloromethane (2 mL) were added. The mixture was cooled to 0°C, and then 1 mL of trifluoroacetic acid (TFA) was added. The mixture was then stirred at 0°C for 3 hours. The solvent was then evaporated under reduced pressure to obtain tert-butyl=2-(N-triduteriomethylamino)acetate TFA salt (320 mg, 2.02 mmol, 100% yield) as a light brown oily substance. This was used directly in the next step. ES LC-MS m / z = 149 (M+H + ).

[0390] Step 5: Synthesis of tert-butyl=2-(N-triduteriomethylcyanamide)acetate JPEG2026048846000114.jpg12128 In a round-bottom flask equipped with a stirring bar, tert-butyl=2-(N-triduteriomethylamino)acetate TFA salt (320 mg, 2.02 mmol), potassium carbonate (837 mg, 6.06 mmol), and acetonitrile (10 mL) were added. The mixture was stirred at room temperature for 0.5 hours, and then a solution of cyanogen bromide (235 mg, 2.22 mmol) in acetonitrile (2 mL) was added. The reaction mixture was stirred at room temperature overnight. The solvent was then removed by decantation, leaving an insoluble residue. The solvent was evaporated under reduced pressure and then purified by flash chromatography using petroleum ether-ethyl acetate (gradient from 0% to 40% ethyl acetate) to obtain tert-butyl=2-(N-triduteriomethylcyanamide)acetate (260 mg, 1.50 mmol, yield 74%) as a white solid. ES LC-MS m / z = 174 (M+H + ) & 196 (M+Na + ).

[0391] Step 6: Synthesis of tert-butyl=2-(2-hydroxy-1-triduteriomethylguanidino)acetate JPEG2026048846000115.jpg17128 In a round-bottom flask equipped with a stirring bar, tert-butyl=2-(N-triduteriomethylcyanamide) acetate (260 mg, 1.50 mmol) and tetrahydrofuran ( 5 mL was added. To this mixture, hydroxylamine (50% aqueous solution, 495 mg, 7.50 mmol) was added. After 1 hour, 10 mL of water was added, and the mixture was extracted three times with 5 mL of dichloromethane. The organic layer was dried over MgSO4, filtered, and concentrated under reduced pressure to obtain tert-butyl=2-(2-hydroxy-1-triduteriomethylguanidino)acetate (298 mg, 1.45 mmol, yield 97%) as a white solid. This was immediately used in the next step. ES LC-MS m / z = 207 (M+H + ).

[0392] Step 7: Synthesis of tert-butyl=2-(3-(tert-butoxycarbonyl)-2-hydroxy-1-triduteriomethylguanidino)acetate JPEG2026048846000116.jpg17128 In a round-bottom flask equipped with a stirring bar, tert-butyl=2-(2-hydroxy-1-triduteriomethylguanidino)acetate (298 mg, 1.45 mmol) and tetrahydrofuran (5 mL) were added. Di-tert-butyl bicarbonate (316 mg, 1.72 mmol) was added to this mixture. The reaction mixture was stirred overnight at room temperature. The solvent was then evaporated under reduced pressure, and the product was purified by flash chromatography using dichloromethane-ethyl acetate (gradient from 0% to 30% ethyl acetate) to obtain tert-butyl=2-(3-(tert-butoxycarbonyl)-2-hydroxy-1-triduteriomethylguanidino)acetate (90 mg, 0.29 mmol, yield 20%) as a white solid. ES LC-MS m / z = 307 (M+H + ). 1¹H NMR (dimethyl sulfoxide-d) δ: 5.71 (s, 2H), 3 .81 (s, 2H), 1.43 (s, 9H),1.41 (s, 9H).

[0393] Other creatine prodrugs and their derivatives can be synthesized using the procedure described above and by selecting appropriate starting materials.

[0394] Example 26: Synthesis of ethyl=2-[methyl-(5-oxo-4H-1,2,4-oxadiazole-3-yl)amino]acetate JPEG2026048846000117.jpg19128 Ethyl 2-[cyano(methyl)amino]acetate (852 mg, 6.0 mmol) and tetrahydrofuran (30 mL) were added to a round-bottom flask equipped with a stirring bar and a nitrogen inlet tube. Hydroxylamine hydrochloride (2.1 g, 30.0 mmol) and triethylamine (1.3 mL, 9.0 mmol) were added to this mixture. After 18 hours, carbonyldiimide (5.8 g, 36.0 mmol) was added, and the reaction mixture was allowed to stand for 1 hour. The solvent was removed by decantation, leaving an insoluble residue. The solvent was evaporated under reduced pressure, and the product was purified by elution using reverse-phase chromatography with water-acetonitrile modified with 0.1% trifluoroacetic acid, respectively. This yielded ethyl 2-[methyl-(5-oxo-4H-1,2,4-oxadiazole-3-yl)amino]acetate as a white solid: 80 mg, 0.40 mmol, yield 7%. ES LC-MS m / z = 202 (M+H + ). 1 1H NMR (chloroform-d) δ: 4.25 (q, J = 7.2 Hz, 2H), 3.96 (s, 2H), 3.04 (s, 3 H), 1.31 (t, J = 7.1 Hz, 3H).Melting point 120-123℃.

[0395] Ethyl 2-[methyl-(5-oxo-4H-1,2,4-oxadiazole-3-yl)amino]acetate can also be synthesized using the procedure described in Kitamure et al, Chem. Pharm. Bull., 2001, 49(3) 268-277.

[0396] Example 27: Synthesis of 2-[methyl-(5-oxo-4H-1,2,4-oxadiazole-3-yl)aminoacetic acid] JPEG2026048846000118.jpg16128 Ethyl 2-[methyl-(5-oxo-4H-1,2,4-oxadiazole-3-yl)amino]acetate (20 mg, 0.1 mmol), tetrahydrofuran (5 mL), and water (5 mL) were added to a round-bottom flask equipped with a stirring bar and a nitrogen inlet tube. Lithium hydroxide monohydrate (4 mg, 0.1 mmol) was added to this mixture. After 1 hour, the solvent was evaporated under reduced pressure, and the product was purified by elution using reverse-phase chromatography with water-acetonitrile modified with 0.1% trifluoroacetic acid. This yielded 2-[methyl-(5-oxo-4H-1,2,4-oxadiazole-3-yl)amino]acetic acid as a white solid: 15 mg, 0.09 mmol, 90% yield. ES LC-MS m / z = 174 (M+H + ). 1 1H NMR (methanol-d4) ) δ: 3.97 (s, 2H), 2, 96 (s,3H).Melting point 150-155℃.

[0397] Other creatine prodrugs and their derivatives can be synthesized using the procedure described above and by selecting appropriate starting materials.

[0398] Example 28: Synthesis of alkyl=2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetate JPEG2026048846000119.jpg52132 Step 1: Synthesis of methyl-2-[ethoxycarbonylcarbamate oil (triduteriomethyl)amino]acetate JPEG2026048846000120.jpg17128 Ethyl N-(thioxomethylene)carbamate (4.1 mL, 35.0 mmol) and dichloromethane (300 mL) were added to a round-bottom flask equipped with a stirring bar and a nitrogen inlet tube. The mixture was cooled to 0°C, and methyl 2-(triduteriomethylamino)acetate trifluoroacetate (7.70 g, 35.0 mmol) was added, followed by triethylamine (4.9 mL, 35.0 mmol). The mixture was stirred overnight while being heated to room temperature. The mixture was washed with 1N HCl (100 mL), dried, and evaporated under reduced pressure. The product was purified by chromatography using a 120 g silica cartridge with heptane-ethyl acetate (gradient from 0 to 30% ethyl acetate). This yielded methyl-2-[ethoxycarbonylcarbamate (triduteriomethyl)amino]acetate: 8.0 g, 33.8 mmol, 96% yield. ES LC-MS m / z = 238 (M+H + ). 1 ¹H NMR (chloroform-d) δ: 7.38 (br s, 1H), 4.41-4.59 (m , 2H), 4.20 (q, J = 7.1 Hz, 2H), 3.80(s, 3H) , 1.31 (t, J = 7.1 Hz, 3H).

[0399] Step 2: Synthesis of methyl=2-[[(Z)-N-ethoxycarbonyl-C-methylsulfanyl-carbonimidoyl]-(triduteriomethyl)amino]acetate JPEG2026048846000121.jpg19128 In a round-bottom flask equipped with a stirrer and a nitrogen inlet tube, [ethoxycarbonylcarbamote oil (triduteriomethyl)amino]acetate (7.82 g, 33.0 mmol), methyl iodide (4.1 mL, 66.0 mmol), and tetrahydrofuran (200 mL) were added. Sodium hydride (60% in oil; 1.32 g, 33.0 mmol) was added to this mixture at room temperature. After 1 hour, the product was added to saturated ammonium chloride solution (100 mL), the aqueous phase was extracted with ethyl acetate (3 × 100 mL), dried to (Na₂SO₄), and the solvent was evaporated under reduced pressure. The product was purified by chromatography using a 120 g silica cartridge with heptane-ethyl acetate (gradient from 0 to 40% ethyl acetate). This yielded methyl=2-[[(Z)-N-ethoxycarbonyl-C-methylsulfanyl-carbonimidoyl]-(triduteriomethyl)amino]acetate: 8.0 g, 32.0 mmol, 97% yield. ES LC-MS m / z = 252 (M+H + ). 1 1H NMR (chloroform-d)δ: 4.30 (s, 2H), 4.16 (q,J = 7.1 Hz, 2H), 3.77 (s, 3H), 2.42 (s, 3H), 1.30 (t,J = 7.1 Hz, 3 H).

[0400] Step 3: Synthesis of methyl=2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetate JPEG2026048846000122.jpg20128 Methyl 2-[[(Z)-N-ethoxycarbonyl-C-methylsulfanyl-carbonimidoyl]-(triduteriomethyl)amino]acetate (7.53 g, 30.0 mmol) and pyridine (50 mL) were added to a round-bottom flask equipped with a stirring bar, a Vigreux column, and a nitrogen inlet tube. Hydroxylamine hydrochloride (2.09 g, 30.0 mmol) was added to this mixture, and the mixture was heated at 60°C for 1 hour. The solvent was evaporated under reduced pressure. Ethyl acetate (100 mL) and water (100 mL) were added. The phases were separated, and the aqueous phase was extracted with ethyl acetate (3 × 100 mL). The organic phases were combined and dried (Na₂SO₄), and the solvent was evaporated under reduced pressure. The product was purified by chromatography using a 120 g silica cartridge with heptane-ethyl acetate (gradient from 0 to 100% ethyl acetate). This yielded methyl=2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetate as a white solid: 3.0 g, 15.8 mmol, yield 53%. ES LC-MS m / z = 191 (M+H + ). 1 1H NMR (chloroform-d)δ: 3.98 (s, 2H), 3.79 (s,3H).Melting point 120-125℃.

[0401] Step 4: Synthesis of 2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetic acid JPEG2026048846000123.jpg20128 In a round-bottom flask equipped with a stirring bar and a nitrogen inlet tube, methyl 2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetate (95 mg, 0.5 mmol), tetrahydrofuran (5 mL), and water (5 mL) were added. Lithium hydroxide monohydrate (21 mg, 0.5 mmol) was added to this mixture. After 1 hour, the solvent was evaporated under reduced pressure, and the product was purified by elution using reverse-phase chromatography with water-acetonitrile modified with 0.1% trifluoroacetic acid. This yielded 2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetic acid as a white solid: 50 mg, 0.28 mmol, yield 57%. ES LC-MS m / z = 177 (M+H + ). 1 1H NMR (methanol-d4) δ: 3.98 (s, 2H). Melting point 1 50-155℃.

[0402] Step 5A: Synthesis of heptyl=2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetate JPEG2026048846000124.jpg20128 2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetic acid (88 mg, 0.50 mmol), 1-heptanol (58 mg, 0.50 mmol), dimethylaminopyridine (92 mg, 0.75 mmol), and dichloromethane (10 mL) were added to a scintillation vial equipped with a stirring bar. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (144 mg, 0.75 mmol) was added to this mixture. After 1 hour, triethylamine (0.07 mL, 0.50 mmol) was added. The reaction mixture was left to stand for another hour. The solvent was evaporated under reduced pressure, and the product was purified by elution using reverse-phase chromatography with water-acetonitrile modified with 0.1% trifluoroacetic acid, respectively. This yielded heptyl=2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetate as a white solid: 70 mg, 0.25 mmol, yield 50%. ES LC-MS m / z = 275 (M+H + ). 1 ¹H NMR (chloroform-d) δ: 11.06 (br s, 1H), 4.17 ( t, J = 6.8 Hz, 2H), 3.95 (s, 2H), 1.56-1.82( m, 4H), 1.19-1.44 (m, 7H), 0.85-0.94(m, 2H) Melting point: 110-115°C.

[0403] Step 5B: Synthesis of ethyl=2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetate JPEG2026048846000125.jpg18128 2-[(5-oxo-2H-1,2,4-oxadiazole-3-yl)-(triduteriomethyl)amino]acetic acid (176 mg, 1.00 mmol), ethanol (46 mg, 1.00 mmol), dimethylaminopyridine (183 mg, 1.50 mmol), and dichloromethane (20 mL) were added to a scintillation vial equipped with a stirring bar. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (288 mg, 1.50 mmol) was added to this mixture. After 1 hour, triethylamine (0.14 mL, 1.00 mmol) was added. The reaction mixture was left to stand for another hour. The solvent was evaporated under reduced pressure, and the pr...

Claims

1. A compound of formula (VII), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof: The compound of formula (VII) is as follows: During the ceremony: R is -CH3 or -CD3; Each R14 independently consists of hydrogen, C1-12 alkyl, substituted C1-12 alkyl, C1-12 heteroalkyl, substituted C1-12 heteroalkyl, C3-12 cycloalkyl, substituted C3-12 cycloalkyl, C4-20 cycloalkylalkyl, substituted C4-20 cycloalkylalkyl, C4-20 heterocycloalkylalkyl, substituted C4-20 heterocycloalkylalkyl, C5-12 aryl, substituted C5-12 aryl, C5-12 heteroaryl, substituted C5-12 heteroaryl, C6-20 arylalkyl, substituted C6-20 heteroarylalkyl, substituted C6-20 heteroarylalkyl, -CH(OR5), -C(O)R5, -C(O)OR5 , or -C(O)(NR3R4); Each R3 and R4 is independently hydrogen, a C1-12 alkyl, or a substituted C1-12 alkyl; and R5 is hydrogen, C1-12 alkyl, substituted C1-12 alkyl, C1-12 heteroalkyl, substituted C1-12 heteroalkyl, C3-12 cycloalkyl, substituted C3-12 cycloalkyl, C4-20 cycloalkylalkyl, substituted C4-20 cycloalkylalkyl, C4-20 heterocycloalkylalkyl, substituted C4-20 heterocycloalkylalkyl, C5-12 aryl, substituted C5-12 aryl, C5-12 heteroaryl, substituted C5-12 heteroaryl, C6-20 arylalkyl, substituted C6-20 arylalkyl, C6-20 heteroarylalkyl, or substituted C6-20 heteroarylalkyl. The aforementioned compound.

2. The compound according to claim 1, wherein R5 is a C1-6 alkyl, a substituted C1-6 alkyl, a C3-7 cycloalkyl, a substituted C3-7 cycloalkyl, a C5-7 aryl, or a substituted C5-7 aryl.

3. The compound according to claim 1, wherein R5 is hydrogen, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-dimethoxyethyl, 1,1-diethoxyethyl, phenyl, 4-methoxyphenyl, benzyl, phenethyl, styryl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 2-pyridyl, 3-pyridyl, or 4-pyridyl.

4. The compound according to claim 1, wherein R5 is hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, sec-pentyl, neopentyl, dodecyl, 1,1-dimethoxyethyl, 1,1-diethoxyethyl, phenyl, cyclohexyl, or 3-pyridyl.

5. The compound according to claim 1, wherein R5 is hydrogen, methyl, ethyl, n-propyl, isopropyl, dodecyl, tert-butyl, phenyl, or cyclohexyl.

6. The compound according to claim 1, wherein R5 is ethyl, isopropyl, or dodecyl.

7. The compound according to claim 1, wherein each R3 and R4 is independently hydrogen.

8. The compound according to claim 1, wherein each substituent is independently a halogen, -NO2, -OH, -NH2, -CN, -CF3, -OCF3, =O, C1-12 alkyl, substituted C1-12 alkyl, C1-12 alkoxy, or substituted C1-12 alkoxy, -COOR10', where R10' is hydrogen, C1-3 alkyl, or -(NR11')2, where each R11' is independently hydrogen or C1-3 alkyl.

9. The compound of formula (VII) is the compound of formula (XXIX), or a pharmaceutically acceptable salt, solvate, tautomer, or stereoisomer thereof; The compound of formula (XXIX) is as follows: In the formula, R is -CH3 or -CD3; and Each R 34 is independently hydrogen, methyl, ethyl, n-propyl, isopropyl, tert-butyl, dodecyl, phenyl, or cyclohexyl. The compound according to claim 1.

10. Hereinafter, or A compound according to claim 1, or a pharmaceutically acceptable salt thereof, selected from the above.

11. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 10 in a therapeutically effective amount, and a pharmaceutically acceptable vehicle.

12. The pharmaceutical composition according to claim 11, comprising one or more sustained-release oral dosage forms.

13. The pharmaceutical composition according to claim 11, wherein the at least one compound is present in an amount effective for treating a patient's disease, the disease being ischemia, oxidative stress, neurodegenerative disease, ischemia-reperfusion injury, cardiovascular disease, genetic disease affecting the creatine kinase system, multiple sclerosis, mental disorders, and muscle fatigue; present in an amount sufficient to bring energy homeostasis to a diseased tissue or organ; present in an amount effective for improving the patient's muscle strength; present in an amount effective for improving the viability of a tissue or organ; or present in an amount effective for improving cell viability.

14. The pharmaceutical composition according to claim 11, wherein at least one compound is present in an amount effective for treating a genetic disease affecting the creatine kinase system.

15. The pharmaceutical composition according to claim 11, wherein at least one compound is present in an amount effective for treating creatine transporter dysfunction.

16. The pharmaceutical composition according to claim 11, wherein at least one compound is present in an amount effective for treating creatine synthesis disorder.

17. A pharmaceutical composition according to claim 11 for treating a disease, wherein the disease is ischemia, oxidative stress, neurodegenerative disease, ischemia-reperfusion injury, cardiovascular disease, genetic disease affecting the creatine kinase system, multiple sclerosis, mental disorder, or muscle fatigue.

18. The pharmaceutical composition according to claim 17, wherein the genetic disease affecting the creatine kinase system is a creatine transporter disorder or a creatine synthesis disorder.

19. The pharmaceutical composition according to claim 11 for treating a genetic disease affecting the creatine kinase system.

20. The pharmaceutical composition according to claim 11 for treating creatine transporter disorders.

21. The pharmaceutical composition according to claim 11 for treating creatine synthesis disorder.

22. The pharmaceutical composition according to claim 11 for improving muscle strength.

23. The pharmaceutical composition according to claim 11 for improving the viability of cells.

24. The pharmaceutical composition according to claim 11 for improving the viability of a tissue or organ.

25. The pharmaceutical composition according to claim 11 for providing energy homeostasis to a tissue or organ.

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