阅读说明：本技术 用于骨骼肌调节的raf-1激酶抑制剂化合物、其方法和用途 (RAF-1 kinase inhibitor compounds for skeletal muscle modulation, methods and uses thereof ) 是由 D·M·巴伦 J·费热 S·卡拉兹 J·米肖 Y·拉蒂诺 P·斯图尔萨茨 于 2019-09-12 设计创作，主要内容包括：本发明涉及用于通过调节肌肉干细胞来改善骨骼肌再生以维持或增加肌肉功能和/或肌肉质量的新型Raf-1激酶抑制剂化合物。例如,本发明可用于要促进肌肉修复的个体和/或经受恶病质前期、恶病质、肌肉减少症、肌病、营养不良和/或肌肉损伤或手术后的恢复的个体。(The present invention relates to novel Raf-1 kinase inhibitor compounds for use in improving skeletal muscle regeneration by modulating muscle stem cells to maintain or increase muscle function and/or muscle mass. For example, the invention may be used in individuals who are to promote muscle repair and/or who are experiencing pre-cachexia, sarcopenia, a myopathy, a malnutrition and/or a muscle injury or post-operative recovery.)

1. A compound of formula (I), or an isomer or salt thereof, for maintaining or increasing skeletal muscle function and/or mass, and/or substantially preventing or reducing muscle atrophy in a subject, the compound represented by formula (I):

wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl.

2. The compound according to claim 1, or an isomer or salt thereof, for maintaining or increasing skeletal muscle function and/or mass and/or substantially preventing or reducing muscle atrophy in an individual, represented by formula (III):

wherein R2, R3, R4 and R7 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl.

3. A compound according to claim 1 or claim 2, wherein the compound is 3- (3, 5-dibromo-4-hydroxy-benzylidene) -5-iodo-1, 3-dihydro-indol-2-one

Or isomers or salts thereof.

4. A compound according to any one of claims 1 to 3, which maintains or increases muscle function and/or mass in an individual by modulating muscle stem cell function, and/or substantially prevents or reduces muscle atrophy in an individual.

5. A composition comprising a compound according to any one of claims 1 to 4 for use in maintaining or increasing muscle function and/or mass and/or substantially preventing or reducing muscle atrophy in an individual.

6. The pharmaceutical composition according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, for use in maintaining or increasing muscle function and/or mass and/or substantially preventing or reducing muscle atrophy in an individual.

7. The pharmaceutical composition according to any one of claims 1 to 6 for use in the prevention or treatment of cachexia or pre-cachexia; sarcopenia, myopathy, malnutrition and/or recovery after muscle injury or surgery.

8. The pharmaceutical composition according to claim 7, wherein cachexia is associated with a disease selected from cancer, chronic heart failure, renal failure, chronic obstructive pulmonary disease, AIDS, autoimmune disorders, chronic inflammatory disorders, cirrhosis of the liver, anorexia, chronic pancreatitis, metabolic acidosis, and/or neurodegenerative diseases.

9. The pharmaceutical composition according to claim 7 or 8, wherein cachexia or pre-cachexia is associated with cancer.

10. The pharmaceutical composition according to claim 9, wherein the treatment of cachexia is associated with a cancer selected from pancreatic cancer, esophageal cancer, gastric cancer, intestinal cancer, lung cancer and/or liver cancer.

11. A pharmaceutical composition according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, for administration alone or in combination with a therapeutic anti-cancer compound or composition.

12. A method of treating cachexia or pre-cachexia, said method comprising administering to a human or animal subject an effective amount of a compound according to any one of claims 1 to 4 or a composition according to any one of claims 5 to 11 or an isomer or pharmaceutically acceptable salt thereof.

13. A method of treating cachexia or pre-cachexia according to claim 12, wherein cachexia or pre-cachexia is associated with a disease selected from cancer, chronic heart failure, renal failure, chronic obstructive pulmonary disease, AIDS, autoimmune disorders, chronic inflammatory disorders, cirrhosis, anorexia, chronic pancreatitis, metabolic acidosis and/or neurodegenerative diseases.

14. The method of treatment according to claim 13, wherein the treatment of cancer cachexia is associated with a cancer selected from pancreatic cancer, esophageal cancer, gastric cancer, intestinal cancer, lung cancer and/or liver cancer.

15. The method of treatment according to claim 14, wherein the treatment of cancer cachexia is measured by reducing weight loss, preventing weight loss, maintaining weight, or increasing weight.

16. Use of a compound according to any one of claims 1 to 4 or a composition according to any one of claims 5 to 11 in a method of treatment, wherein cancer cachexia is the result of treatment of cancer with a chemotherapeutic agent.

17. Use of a compound according to any one of claims 1 to 4 or a composition according to any one of claims 5 to 11 in combination with a dietary intervention of high calorie, high protein, high carbohydrate, vitamin B3, B12 and/or vitamin D supplement, antioxidant, omega fatty acid and/or polyphenol in a method of preventing or treating cachexia.

18. Use of a compound according to any one of claims 1 to 4 or a composition according to any one of claims 5 to 11 in the manufacture of a medicament for the prevention and/or treatment of cachexia or pre-cachexia, sarcopenia, myopathy, malnutrition and/or recovery following muscle injury or surgery.

19. Use of a compound according to any one of claims 1 to 4 or a composition according to any one of claims 5 to 11 in a non-human animal for optimizing meat production by increasing skeletal muscle mass and/or function.

20. A kit for the prevention or treatment of cachexia or pre-cachexia, said kit comprising a compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof or a composition according to any one of claims 5 to 11 or a pharmaceutically acceptable salt thereof, administered alone or in combination with an anti-cancer treatment.

21. A kit for maintaining or increasing muscle function and/or muscle mass and/or substantially preventing or reducing muscle atrophy in an individual with sarcopenia, myopathy, malnutrition and/or recovery after muscle injury or surgery, the kit comprising a compound according to any one of claims 1 to 4 or a composition according to any one of claims 5 to 11 or a pharmaceutically acceptable salt thereof.

22. The kit of claim 20 or claim 21, wherein the kit further comprises a high calorie, high protein, high carbohydrate, vitamin B3, B12, and/or vitamin D supplement, an antioxidant, a dietary intervention of omega fatty acids and/or polyphenols, and instructions for daily administration.

Technical Field

The present invention relates to novel Raf-1 kinase inhibitor compounds for use in improving skeletal muscle regeneration by modulating muscle stem cells to maintain or increase muscle function and/or muscle mass. For example, the invention may be used in individuals who are to promote muscle repair and/or who are experiencing pre-cachexia, sarcopenia, a myopathy, a malnutrition and/or a muscle injury or post-operative recovery.

Background

Skeletal muscle regeneration is an important mechanism for the repair and maintenance of muscle mass and function throughout life. Skeletal muscle regeneration requires the involvement of mainly myogenic progenitor cells (called muscle stem cells or satellite cells).

Non-proliferating, quiescent satellite cells that border resting skeletal muscle can be identified by their different positions between the sarcolemma and basal lamina, high nuclear to cytoplasmic volume ratios, few organelles (e.g., ribosomes, endoplasmic reticulum, mitochondria, golgi apparatus), small nuclear sizes, and large amounts of heterochromatin relative to the myonucleus. Activated satellite cells, on the other hand, have an increased number of pits (caveolae), cytoplasmic apparatus and reduced levels of heterochromatin.

These muscle satellite cells are part of the adult stem cell nest, and they are involved in the normal growth of muscle, and regeneration after injury or disease. Thus, they are potential targets for enhancing muscle regeneration in both healthy and diseased conditions. Skeletal muscle regeneration follows a series of steps that recapitulate the developmental stage. Muscle progenitor cells must leave a quiescent state, become active, proliferate and participate in myogenic differentiation.

Satellite cells express genetic markers at different stages of myogenesis and proliferation. Pax7 and Pax3 are considered satellite cell markers. For example, activated satellite cells expressing low levels of Pax7 are more prone to differentiation, while high levels of Pax7 are associated with cells that are less prone to differentiation and have more undifferentiated stem cell characteristics. Activation and induction of myogenesis is commonly regulated by myogenic regulatory factors such as MyoD, Myf5, myogenin and MRF 4. Negative regulation of myostatin and TGFb inhibited satellite cell differentiation (Almeida et al, 2016).

Previous experimental therapies that have included myoblast transplantation have not been completely successful because the regenerative potential of more committed and differentiated myoblasts and muscle fibers is lower compared to muscle stem cells.

Thus, there remains a significant need to find compounds, compositions and methods that directly modulate muscle stem cells to maintain muscle health and improve muscle regeneration. Such compounds, compositions, and methods of treatment may help individuals with muscle stem cell dysfunction and/or individuals experiencing muscle diseases and disorders such as cachexia or sarcopenia by promoting the maintenance of increased muscle function and/or muscle mass.

Disclosure of Invention

The present inventors have found novel compounds and compositions for modulating skeletal muscle function and improving skeletal muscle regeneration in order to improve muscle repair after injury or for counteracting muscle atrophy that occurs in a variety of pathological conditions, particularly cachexia and sarcopenia.

In one embodiment, the present invention relates to a compound of formula (I):

wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl.

The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group. The phenyl ring C may be replaced by a pyridyl, pyrimidinyl, naphthyl, quinolinyl or isoquinolinyl group. These groups may be further substituted with: h; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group.

In another embodiment, the invention relates to a compound of formula (II):

wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group.

The phenyl ring C may be replaced by a pyridyl, pyrimidinyl, naphthyl, quinolinyl or isoquinolinyl group. These groups may be further substituted with: h; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group.

In another embodiment, the invention relates to a compound of formula (III):

wherein R2, R3, R4 and R7 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group.

In a preferred embodiment, the invention relates to compounds of formula (III) wherein R2, R4 and R7 are substituted halo.

In a preferred embodiment of the invention, the compound is 3- (3, 5-dibromo-4-hydroxy-benzylidene) -5-iodo-1, 3-dihydro-indol-2-one:

and isomers or salts thereof.

3- (3, 5-dibromo-4-hydroxy-benzylidene) -5-iodo-1, 3-dihydro-indol-2-one also known as GW 5074; 3- (3, 5-dibromo-4-hydroxybenzylidene) -5-iodo-1, 3-indolin-2-one; 3- [ (3, 5-dibromo-4-hydroxyphenyl) methylene group]-1, 3-dihydro-5-iodo-2H-indol-2-one; 3- [ (3, 5-dibromo-4-hydroxyphenyl) methylene group]-5-iodo-1H-indol-2-one; 8.5-iodo-3- [ (3, 5-dibromo-4-hydroxyphenyl) methylene]-2-indolinone; (Z) -3- (3, 5-dibromo-4-hydroxybenzylidene) -5-iodoindolin-2-one; CAS number 220904-83-6, having formula C₁₅H₈Br₂INO₂And a molecular weight of 520.946 g/mol.

The compounds and compositions of the invention are useful for modulating muscle stem cell function to maintain or increase skeletal muscle function and/or mass in an individual, and/or to substantially prevent or reduce muscle atrophy in an individual. In particular, to enhance: the number of muscle stem cells, the function of the muscle stem cells, myogenesis and muscle growth.

The compounds and compositions of the present invention are useful for promoting muscle regeneration, recovering from muscle atrophy or muscle damage, and/or preventing or treating sarcopenia or cachexia; or pre-cachexia. In particular, wherein sarcopenia is a loss of muscle mass and/or strength associated with aging and cachexia is associated with a disease, for example when associated with cancer, chronic heart failure, renal failure, chronic obstructive pulmonary disease, AIDS, autoimmune disorders, chronic inflammatory disorders, liver cirrhosis, anorexia, chronic pancreatitis, metabolic acidosis and/or neurodegenerative diseases (Von Haehling et al, 2014).

Drawings

FIG. 1 proliferation and myogenic orientation of muscle stem cells using GW5074

Figure 1 shows that GW5074 promotes expansion of human primary muscle stem cells as evidenced by an increase in Pax7+ cells and promotes their myogenic orientation as evidenced by an increase in the proportion of MyoD + cells.

Figure 1A shows the amount of differentiated myoblast (MyoD +) cells expressed as a percentage of total cells in a dose-dependent response to compound GW 5074. MyoD + cells represent cells that do not express Pax7 and only MyoD.

Figure 1B shows the number of Pax7+ cells in a dose-dependent response to compound GW 5074. Pax7+ cells represent cells expressing Pax7, independent of MyoD expression.

Indicates differences from control, one-way anova p <0.01, data expressed as mean +/-SEM

FIG. 2-muscle fiber growth and differentiation using GW5074

Figure 2 shows that GW5074 enhances myogenic differentiation and myofiber growth of human primary muscle cells.

Figure 2A shows myogenic differentiation measured by fusion factors as the percentage of nuclei within troponin T positive myotubes in a dose-dependent response to compound GW 5074.

Figure 2B shows muscle fiber growth, expressed as the area covered by troponin T positive myotubes in a dose-dependent response to compound GW 5074.

Indicates differences from control, one-way anova p <0.01, data expressed as mean +/-SEM

Detailed Description

Preferred features and embodiments of the present invention will now be described by way of non-limiting examples.

Compounds of the invention

The compounds of the present invention are Raf-1 inhibitors. Raf-1 is also known as c-Raf, a member of the proto-oncogene serine/threonine protein kinase family.

In one embodiment, the present invention relates to a compound of formula (I):

wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl.

The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group. The phenyl ring C may be replaced by a pyridyl, pyrimidinyl, naphthyl, quinolinyl or isoquinolinyl group. These groups may be further substituted with: h; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group.

In another embodiment, the invention relates to a compound of formula (II):

wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group.

The phenyl ring C may be replaced by a pyridyl, pyrimidinyl, naphthyl, quinolinyl or isoquinolinyl group. These groups may be further substituted with: h; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group.

In another embodiment, the invention relates to a compound of formula (III):

wherein R2, R3, R4 and R7 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids. The alkyl, alkenyl or alkynyl chain may be substituted with one or two sulfur atoms contained in groups such as mercapto and thioether. The alkyl, alkenyl or alkynyl chain may be terminated by a cyanide group.

In a preferred embodiment, compounds of formula (III) wherein R2, R4 and R7 are substituted halo.

In a preferred embodiment of the invention, the compound is 3- (3, 5-dibromo-4-hydroxy-benzylidene) -5-iodo-1, 3-dihydro-indol-2-one:

and isomers or salts thereof.

3- (3, 5-dibromo-4-hydroxy-benzylidene) -5-iodo-1, 3-dihydro-indol-2-one also known as GW 5074; 3- (3, 5-dibromo-4-hydroxybenzylidene) -5-iodo-1, 3-indolin-2-one; 3- [ (3, 5-dibromo-4-hydroxyphenyl) methylene group]-1, 3-dihydro-5-iodo-2H-indol-2-one; 3- [ (3, 5-dibromo-4-hydroxyphenyl) methylene group]-5-iodo-1H-indol-2-one; 8.5-iodo-3- [ (3, 5-dibromo-4-hydroxyphenyl) methylene]-2-indolinone; (Z) -3- (3, 5-dibromo-4-hydroxybenzylidene) -5-iodoindolin-2-one; CAS number 220904-83-6, having formula C₁₅H₈Br₂INO₂And a molecular weight of 520.946 g/mol.

In another embodiment of the invention, isomers of 3- (3, 5-dibromo-4-hydroxy-benzylidene) -5-iodo-1, 3-dihydro-indol-2-one and salts thereof are also embodied by the invention.

General chemical nomenclature

The term "alkyl" refers to a branched or straight saturated hydrocarbon chain having 1 to 20 carbon atoms, or 1 to 15 carbon atoms, or 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 4 carbon atoms. The term is exemplified by groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-hexyl, n-decyl, tetradecyl.

The term "substituted alkyl" refers to:

1) an alkyl chain as defined above having 1, 2, 3, 4 or 5 substituents (in some embodiments, 1, 2 or 3 substituents) selected from alkyl; alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, cycloalkoxy, cycloalkenyloxy, acyl, amido, acyloxy, amino, substituted amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -S (O) -alkyl, -S (O) -cycloalkyl, -S (O) -heterocyclyl, -S (O) -aryl, -S (O) -heteroaryl, -S (O) 2-alkyl, -S (O) 2-cycloalkyl, substituted amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, hydroxy, thio, carboxyl, carboxyalkyl, arylthio, heteroarylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, -S (O) 2-heterocyclyl, -S (O) 2-aryl and-S (O) 2-heteroaryl. Unless otherwise constrained by the definition, all substituents may optionally be further substituted by 1, 2 or 3 substituents selected from alkyl, alkenyl, alkynyl, carboxyl, carboxyalkyl, aminocarbonyl, hydroxyl, alkoxy, halogen, CF3, amino, substituted amino, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl and-s (o) n R < a >, wherein R < a > is alkyl, aryl or heteroaryl and n is 0, 1 or 2; or

2) Interrupted by an alkyl chain of 1 to 10 atoms (e.g. 1, 2, 3, 4 or 5 atoms) as defined above, these atoms being independently selected from oxygen, sulphur and NR < a >, wherein R < a > is selected from hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclyl. All substituents may be optionally further substituted with alkyl, alkenyl, alkynyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF3, amino, substituted amino, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl and-s (o) n R < a > wherein R < a > is alkyl, aryl or heteroaryl and n is 0, 1 or 2; or

3) An alkyl chain as defined above having 1, 2, 3, 4 or 5 substituents as defined above and further interrupted by 1 to 10 atoms (e.g. 1, 2, 3, 4 or 5 atoms) as defined above.

The term "alkenyl" refers to a class of alkyl chains in which two atoms of the alkyl group form a double bond that is not part of an aromatic group. That is, the alkenyl chain comprises the pattern R-c (R) ═ c (R) -R, where R refers to the remainder of the alkenyl group, which may be the same or different. Non-limiting examples of alkenyl chains include-CH ═ CH2, -C (CH3) ═ CH2, -CH ═ CHCH3, -C (CH3) ═ CHCH3, -CH2-CH ═ C (CH3)2, and-C (CH3)2-CH ═ CH 2. The alkenyl moiety may be branched, straight-chain or cyclic (in which case it will also be referred to as a "cycloalkenyl" group).

The term "substituted alkenyl" refers to:

1) alkenyl chains as defined above having 1, 2, 3, 4, or 5 substituents (in some embodiments, 1, 2, or 3 substituents) selected from alkyl; alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, cycloalkoxy, cycloalkenyloxy, acyl, amido, acyloxy, amino, substituted amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -S (O) -alkyl, -S (O) -cycloalkyl, -S (O) -heterocyclyl, -S (O) -aryl, -S (O) -heteroaryl, -S (O) 2-alkyl, -S (O) 2-cycloalkyl, substituted amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, hydroxy, thio, carboxyl, carboxyalkyl, arylthio, heteroarylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, -S (O) 2-heterocyclyl, -S (O) 2-aryl and-S (O) 2-heteroaryl. Unless otherwise constrained by the definition, all substituents may optionally be further substituted by 1, 2 or 3 substituents selected from alkyl, alkenyl, alkynyl, carboxyl, carboxyalkyl, aminocarbonyl, hydroxyl, alkoxy, halogen, CF3, amino, substituted amino, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl and-s (o) n R < a >, wherein R < a > is alkyl, aryl or heteroaryl and n is 0, 1 or 2; or

2) An alkenyl chain interrupted by 1 to 10 atoms (e.g. 1, 2, 3, 4 or 5 atoms) as defined above, independently selected from oxygen, sulphur and NR < a >, wherein R < a > is selected from hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclyl. All substituents may be optionally further substituted with alkyl, alkenyl, alkynyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF3, amino, substituted amino, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl and-s (o) n R < a > wherein R < a > is alkyl, aryl or heteroaryl and n is 0, 1 or 2; or

3) An alkenyl chain as defined above having 1, 2, 3, 4 or 5 substituents as defined above and further interrupted by 1 to 10 atoms (e.g. 1, 2, 3, 4 or 5 atoms) as defined above.

The term "alkynyl" refers to a class of alkyl chains in which two atoms of the alkyl group form a triple bond. That is, alkynyl groups contain the pattern R-C ≡ C-R, where R refers to the remainder of the alkynyl group, which may be the same or different. Non-limiting examples of alkynyl groups include-C ≡ CH, -C ≡ CCH3 and-C ≡ CCH2CH 3. The "R" moiety of the alkynyl moiety may be branched, straight chain or cyclic.

The term "substituted alkynyl" refers to:

1) an alkynyl chain as defined above having 1, 2, 3, 4 or 5 substituents (in some embodiments, 1, 2 or 3 substituents) selected from alkyl; alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, cycloalkoxy, cycloalkenyloxy, acyl, amido, acyloxy, amino, substituted amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -S (O) -alkyl, -S (O) -cycloalkyl, -S (O) -heterocyclyl, -S (O) -aryl, -S (O) -heteroaryl, -S (O) 2-alkyl, -S (O) 2-cycloalkyl, substituted amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, hydroxy, thio, carboxyl, carboxyalkyl, arylthio, heteroarylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, -S (O) 2-heterocyclyl, -S (O) 2-aryl and-S (O) 2-heteroaryl. Unless otherwise constrained by the definition, all substituents may optionally be further substituted by 1, 2 or 3 substituents selected from alkyl, alkenyl, alkynyl, carboxyl, carboxyalkyl, aminocarbonyl, hydroxyl, alkoxy, halogen, CF3, amino, substituted amino, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl and-s (o) n R < a >, wherein R < a > is alkyl, aryl or heteroaryl and n is 0, 1 or 2; or

2) An alkynyl chain as defined above interrupted by 1 to 10 atoms (e.g. 1, 2, 3, 4 or 5 atoms) independently selected from oxygen, sulphur and NR < a >, wherein R < a > is selected from hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclyl. All substituents may be optionally further substituted with alkyl, alkenyl, alkynyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF3, amino, substituted amino, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl and-s (o) n R < a > wherein R < a > is alkyl, aryl or heteroaryl and n is 0, 1 or 2; or

3) An alkynyl chain as defined above having 1, 2, 3, 4 or 5 substituents as defined above and further interrupted by 1 to 10 atoms (e.g. 1, 2, 3, 4 or 5 atoms) as defined above.

As used herein, the term "ring" refers to any covalently closed structure. Rings include, for example, carbocycles (e.g., aryl and cycloalkyl), heterocycles (e.g., heteroaryl and non-aromatic heterocycle), aromatics (e.g., aryl and heteroaryl), and non-aromatics (e.g., cycloalkyl and non-aromatic heterocycle). The ring may be optionally substituted. The ring may form part of a ring system. As used herein, the term "ring system" refers to two or more rings, wherein two or more rings are fused. The term "fused" refers to a structure in which two or more rings share one or more bonds.

The term "halogen atom" may refer to a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

The term "naphthalene" refers to a structure consisting of a pair of fused benzene rings.

As used herein, the term "analog" is understood to mean a compound having a structure similar to another structure, but differing therefrom in certain components. A "derivative" is a compound that can be imagined to be produced or actually synthesized from the parent compound by substituting one or more atoms with another atom or group of atoms.

As used herein, the term "isomer" is understood to mean a compound having the same molecular formula but having a different arrangement of atoms in the molecule.

Salts refer in particular to pharmaceutically acceptable salts of the compounds of formula I, II or III.

Such salts are formed, for example, from compounds of formula I having a basic nitrogen atom, preferably with organic or inorganic acids, as acid addition salts, especially pharmaceutically acceptable salts. Suitable inorganic acids are, for example, hydrohalic acids, such as hydrochloric acid, sulfuric acid or phosphoric acid. Suitable organic acids are, for example, carboxylic, phosphonic, sulfonic or sulfamic acids, for example acetic acid, propionic acid, octanoic acid, decanoic acid, dodecanoic acid, glycolic acid, lactic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, malic acid, tartaric acid, citric acid, amino acids such as glutamic acid or aspartic acid, maleic acid, hydroxymaleic acid, methylmaleic acid, cyclohexanecarboxylic acid, adamantanecarboxylic acid, benzoic acid, salicylic acid, 4-aminosalicylic acid, phthalic acid, phenylacetic acid, mandelic acid, cinnamic acid, methanesulfonic acid or ethanesulfonic acid, 2-hydroxyethanesulfonic acid, ethane-1, 2-disulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 1, 5-naphthalenedisulfonic acid, 2-methylbenzenesulfonic acid, 3-methylbenzenesulfonic acid or 4-methylbenzenesulfonic acid, methylsulfuric acid, ethylsulfuric acid, Dodecyl sulfuric acid, N-cyclohexyl sulfamic acid, N-methyl-sulfamic acid, N-ethyl-sulfamic acid or N-propyl-sulfamic acid, or other organic protic acids, such as ascorbic acid.

In the presence of negatively charged groups such as carboxy or sulfo, salts may also be formed with bases, for example metal or ammonium salts, such as alkali metal or alkaline earth metal salts, for example sodium, potassium, magnesium or calcium salts, or ammonium salts using ammonia or suitable organic amines, such as tertiary monoamines, for example triethylamine or tris (2-hydroxyethyl) amine, or heterocyclic bases, for example N-ethyl-piperidine or N, N' -dimethylpiperazine.

When a basic group and an acidic group are present in the same molecule, the compounds of formula I may also form internal salts.

For isolation or purification purposes, it is also possible to use pharmaceutically unacceptable salts, such as picrates or perchlorates. For therapeutic use, only pharmaceutically acceptable salts or free compounds are used (where applicable in the form of pharmaceutical preparations), and these are therefore preferred.

Any reference to free compounds hereinbefore and hereinafter should be understood as referring also to the corresponding salts, as appropriate and expedient, in view of the close relationship between the novel compounds in free form and those in the form of their salts, including those salts which may be used as intermediates, for example, in the purification or identification of the novel compounds.

In one embodiment of the present invention, there is provided a compound of formula (I), or an isomer or salt thereof, to maintain or increase skeletal muscle function and/or mass, and/or substantially prevent or reduce muscle atrophy in a subject, the compound being represented by formula (I):

wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl.

In another embodiment of the present invention, there is provided a compound of formula (II), or an isomer or salt thereof, to maintain or increase skeletal muscle function and/or mass, and/or substantially prevent or reduce muscle atrophy in a subject, the compound being represented by formula (II):

wherein R1, R2, R3, R4, R5, R6, R7, R8, and R9 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl. The alkyl, alkenyl or alkynyl chain may be substituted with one or two oxygen atoms contained in groups such as ethers, primary, secondary and tertiary alcohols, aldehydes and carboxylic acids.

In another embodiment of the present invention, a compound, or an isomer or salt thereof, is provided to maintain or increase skeletal muscle function and/or mass and/or substantially prevent or reduce muscle atrophy in an individual, the compound being represented by formula (III):

wherein R2, R3, R4 and R7 are each independently H; OH; OMe; o-alkyl; SH; S-Me; an S-alkyl group; halogen; primary, secondary or tertiary alcohols, ketones, aldehydes; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; an alkyl cyanide; a nitro group; a sulfonate ester; a sulfate ester; linear, optionally substituted and/or optionally branched C2 to C10 alkenyl; linear, optionally substituted and/or optionally branched C2 to C10 alkynyl.

In a preferred embodiment of the invention, the compound provided for maintaining or increasing skeletal muscle function and/or mass, and/or substantially preventing or reducing muscle atrophy in an individual is 3- (3, 5-dibromo-4-hydroxy-benzylidene) -5-iodo-1, 3-dihydro-indol-2-one

Or isomers or salts thereof.

In one embodiment of the invention, the compounds of the invention modulate muscle stem cell function to maintain or increase skeletal muscle function and/or mass in an individual and/or substantially prevent or reduce muscle atrophy in an individual and/or enhance muscle repair following injury, for example by accelerating repair of muscle fibers or reducing fibrosis and muscle stiffness or reducing muscle fat infiltration.

In another embodiment of the invention, the compounds of the invention modulate muscle stem cell function by proliferation and/or differentiation of skeletal muscle stem cells.

In another embodiment of the invention, the compounds of the invention modulate muscle stem cell function by myogenesis.

Pharmaceutical composition

The pharmaceutical composition comprises, for example, from about 10% to about 100%, preferably from about 20% to about 60% of the active ingredient. Pharmaceutical preparations for parenteral or enteral administration are, for example, those in unit dosage forms, such as sugar-coated tablets, capsules or suppositories, and furthermore ampoules. If not indicated otherwise, these are prepared in a manner known per se, for example by means of conventional mixing, granulating, sugar-coating, dissolving or lyophilizing processes. It will be appreciated that the unit content contained in an individual dose of each dosage form need not in itself constitute an effective amount, since the necessary effective amount can be reached by administration of a plurality of dosage units.

In particular, a therapeutically effective amount of a compound of the invention may be administered simultaneously or sequentially and in any order, and for combinations, the components may be administered separately or as a fixed combination. For example, when used in a method of treatment of cachexia associated with cancer chemotherapy according to the present invention, simultaneous or sequential in any order, administration of a combination partner (a) in free or pharmaceutically acceptable salt form and (ii) administration of a combination partner (b) in free or pharmaceutically acceptable salt form may be included in jointly therapeutically effective amounts, preferably in synergistically effective amounts, e.g. in daily doses corresponding to the amounts described herein.

The individual combination formulations of the present invention may be administered separately at different times during the course of therapy or concurrently in divided or single combination forms. The invention encompasses all such regimens of simultaneous or alternating treatment, and the term "administering" should be construed accordingly.

The effective dosage may vary depending upon the particular compound or pharmaceutical composition employed, the mode of administration, the condition being treated, the severity of the condition being treated. Thus, the dosage regimen will be selected in accordance with a variety of factors including the route of administration and the renal and hepatic function of the patient. A physician, clinician or veterinarian of ordinary skill in the art can readily determine and prescribe the effective amount of the single active ingredients required to prevent, counter or arrest the progress of the condition. Optimal precision in achieving concentration of the active ingredient within the range that yields efficacy without toxicity requires a regimen based on the kinetics of the active ingredient's availability to target sites.

The compound of formula I, II or III can be administered by any route, including orally, parenterally, e.g., intraperitoneally, intravenously, intramuscularly, subcutaneously, intratumorally, or rectally, or parenterally. Preferably, the compound of formula I, II or III is administered orally, preferably at a daily dose of 1mg/kg body weight to 300mg/kg body weight or for most larger primates at a daily dose of 50mg to 5000mg, preferably 500mg to 3000 mg. Preferred oral daily doses are in the range of 1mg/kg body weight to 75mg/kg body weight, or for most larger primates, in the range of 10mg to 2000mg, in a single dose or divided into multiple doses, such as twice daily dosing.

The compound of formula I, II or III is preferably administered orally to a human at a dose in the range of about 100 mg/day to 2000 mg/day, more preferably 500 mg/day to 1500 mg/day, for example 1000 mg/day, and most preferably 750 mg/day to 1500 mg/day.

In one embodiment of the invention, the composition of the invention is provided for use in maintaining or increasing muscle function and/or mass and/or substantially preventing or reducing muscle atrophy in an individual.

In another embodiment of the invention, the composition is a pharmaceutical composition of the invention, or a pharmaceutically acceptable salt thereof, provided for maintaining or increasing muscle function and/or mass and/or substantially preventing or reducing muscle atrophy in an individual.

In another embodiment of the invention, the composition is a pharmaceutical composition comprising a compound of the invention, wherein modulation of muscle stem cell function is measured by an increase in the number of muscle stem cells and/or myoblasts and/or myotubes.

In one embodiment of the invention, a pharmaceutical composition or a pharmaceutically acceptable salt thereof is provided to maintain or increase muscle function and/or mass and/or substantially prevent or reduce muscle atrophy in an individual.

In another embodiment of the present invention, there is provided a pharmaceutical composition or a pharmaceutically acceptable salt thereof for the prevention or treatment of cachexia or pre-cachexia; sarcopenia, myopathy, malnutrition and/or recovery after muscle injury or surgery.

In another embodiment of the present invention, the pharmaceutical composition of the present invention is provided for the prevention or treatment of cachexia, wherein the cachexia is associated with a disease selected from cancer, chronic heart failure, renal failure, chronic obstructive pulmonary disease, AIDS, autoimmune disorders, chronic inflammatory disorders, liver cirrhosis, anorexia, chronic pancreatitis, metabolic acidosis, and/or neurodegenerative diseases.

In a preferred embodiment of the present invention, the pharmaceutical composition of the present invention is provided for use in the prevention or treatment of cancer-related cachexia or pre-cachexia.

In another preferred embodiment of the present invention, the pharmaceutical composition of the present invention is provided for use in the treatment of cachexia associated with a cancer selected from pancreatic cancer, esophageal cancer, gastric cancer, intestinal cancer, lung cancer and/or liver cancer.

In another embodiment of the present invention, there is provided a compound or composition of the present invention for use in the manufacture of a medicament for the prevention and/or treatment of cachexia.

Combination for treating cancer with chemotherapeutic agents

The combinations of the invention comprise at least one compound of the invention and a chemotherapeutic agent for the treatment of cancer, wherein the active ingredients are present in each case in free form or in the form of a pharmaceutically acceptable salt, and optionally at least one pharmaceutically acceptable carrier.

Administration of the combination results in surprising beneficial effects of slowing, arresting or reversing the progression of muscle atrophy, such as less cachexia, improved quality of life and reduced mortality and morbidity, compared to monotherapy administering only one of the pharmaceutically active ingredients.

In one embodiment of the invention, the pharmaceutical composition of the invention, or a pharmaceutically acceptable salt thereof, may be administered in combination with a therapeutic anti-cancer compound.

Multi-part medicine box

A combined preparation can be defined as a "kit of parts" in the sense that it can be dosed independently or by using different fixed combinations with different combined amounts, i.e. simultaneously or at different time points. The parts of the kit of parts can then, for example, be administered simultaneously or chronologically staggered, i.e. at equal or different time intervals at different time points for any part of the kit of parts. Very preferably, the time intervals are chosen such that the effect on the treated disease in the combined use of the parts is larger than the effect which would be obtained by use of only any one of the combination formulations (a) and (b). The ratio of the total amounts of the combination formulation (a) to the combination formulation (b) administered in the combined preparation may be varied, for example in order to cope with the needs of a sub-population of patients to be treated or the needs of individual patients who may need to be different due to the particular disease, age, sex, body weight, etc. of the patients. Preferably, there is at least one benefit, such as a mutual enhancement of the effects of the combined formulation.

In one embodiment of the present invention, there is provided a kit of parts for the prevention or treatment of cachexia or pre-cachexia, comprising a compound or composition of the invention or a pharmaceutically acceptable salt thereof.

In another embodiment of the present invention, a kit of parts for the prevention or treatment of cachexia or pre-cachexia comprising a compound of the present invention or a pharmaceutically acceptable salt thereof administered alone or in combination with an anti-cancer therapy is provided.

In another embodiment of the present invention, there is provided a kit of parts for maintaining or increasing muscle function and/or muscle mass and/or substantially preventing or reducing muscle atrophy in an individual with sarcopenia, myopathy, malnutrition and/or muscle damage or post-operative recovery, comprising a compound or composition of the invention or a pharmaceutically acceptable salt thereof.

In another embodiment of the invention, a kit of parts is provided, wherein the kit further comprises instructions for dietary intervention of high calorie, high protein, high carbohydrate, vitamin B3, B12, and/or vitamin D supplement, antioxidant, omega fatty acid, and/or polyphenol for daily administration.

Combinations of the compounds and compositions of the invention with dietary intervention

The term "dietary intervention" refers to an external factor that is applied to an individual and causes a change in the individual's diet. In one embodiment, the dietary intervention is a high calorie diet. In another embodiment, the dietary intervention is a high protein and/or carbohydrate diet. In another embodiment, the dietary intervention is a diet supplemented with vitamins and minerals, in particular vitamin B12 and/or vitamin D. In another embodiment, the dietary intervention is supplemented with an antioxidant, such as N-acetyl-cysteine. In another embodiment, the dietary intervention is supplemented with omega fatty acids. In another embodiment, the dietary intervention is supplemented with polyphenols or vitamin B3 that increase mitochondrial activity, such as nicotinamide riboside.

The diet may be a diet adapted to the starting body weight of the individual.

The dietary intervention may comprise administering at least one dietary product. The dietary product may be a meal replacement product or a supplement product that may, for example, increase an individual's appetite. The dietary product may include a food product, a beverage, a pet food product, a food supplement, a nutraceutical, a food additive, or a nutritional formula. Exemplary oral nutritional supplements include nestle Boost, Resource, and Meritene products.

In one embodiment of the invention, the compounds or compositions of the invention may be used in a method of preventing or treating cachexia in combination with a high calorie, high protein, high carbohydrate, vitamin B3, vitamin B12 and/or vitamin D supplement, antioxidant, omega fatty acid and/or polyphenol dietary intervention.

Cachexia and related diseases

The present invention provides compounds, compositions and methods for preventing and/or treating cachexia or skeletal muscle wasting syndrome by modulating skeletal muscle stem cells. Cachexia is a complex metabolic syndrome associated with underlying disease and is characterized by muscle loss with or without loss of fat mass. The prominent clinical features of cachexia are adult weight loss (correction of fluid retention) or undersrowth in children (exclusion of endocrine disorders).

Cachexia is often present in patients with diseases such as cancer, chronic heart failure, renal failure, chronic obstructive pulmonary disease, AIDS, autoimmune disorders, chronic inflammatory disorders, cirrhosis, anorexia, chronic pancreatitis, and/or metabolic acidosis, and neurodegenerative diseases.

There are certain types of cancer, where cachexia is particularly prevalent, such as pancreatic, esophageal, gastric, intestinal, lung, and/or liver cancer.

Internationally accepted diagnostic criteria for cachexia are based on current weight and height (body mass index [ BMI ]]＜20kg/m²) Or skeletal muscle mass (measured by DXA, MRI, CT or bioimpedance), greater than 5% weight loss over a limited period of time, e.g. 6 months, or greater than 2% weight loss in individuals who have shown depletion. Cachexia can develop gradually in various stages, i.e., cachexia develops in the early stage and then into intractable cachexia. Severity can be classified according to the extent of consumption of energy storage and body protein (BMI) combined with the extent of sustained weight loss.

In particular, cancer cachexia has been defined as weight loss > 5% (no simple hunger) over the past 6 months; or BMI < 20 and any weight loss > 2%; or the skeletal muscle index of limbs according with sarcopenia (male < 7.26 kg/m)²(ii) a Female with a view to preventing the formation of wrinkles<5·45kg/m²) And any weight loss > 2% (Fearon et al, 2011).

Pre-cachexia can be defined as weight loss ≦ 5% along with anorexia and metabolic changes. Currently, there are no robust biomarkers to identify those pre-cachectic patients who are likely to progress further or the rate at which they will progress further. Refractory cachexia is defined essentially based on the clinical characteristics and condition of the patient.

It is to be understood that the compounds, compositions and methods of the present invention may be beneficial in the prevention and/or treatment of pre-cachexia as well as cachectic conditions, in particular in the maintenance or improvement of skeletal muscle mass and/or muscle function.

In one embodiment of the present invention, the present invention provides a method of treating cachexia or pre-cachexia comprising administering to a human or animal subject an effective amount of a compound of the present invention or an isomer or a pharmaceutically acceptable salt thereof.

In another embodiment of the present invention, there is provided a method of treating cachexia or pre-cachexia comprising administering to a human or animal subject an effective amount of a compound of the present invention or an isomer or a pharmaceutically acceptable salt thereof, wherein the cachexia or pre-cachexia is associated with a disease selected from the group consisting of cancer, chronic heart failure, renal failure, chronic obstructive pulmonary disease, AIDS, autoimmune disorders, chronic inflammatory disorders, liver cirrhosis, anorexia, chronic pancreatitis, metabolic acidosis, and/or neurodegenerative diseases.

In a preferred embodiment of the present invention, the present invention provides a method of treating cancer cachexia associated with a cancer selected from pancreatic, esophageal, gastric, intestinal, lung and/or liver cancer.

In yet another embodiment of the invention, the invention provides a method of treatment wherein the treatment of cancer cachexia is measured by reducing weight loss, preventing weight loss, maintaining weight, or increasing weight.

In another embodiment of the invention, the compounds or compositions of the invention may be used in a method of treatment, wherein cancer cachexia is the result of treatment of cancer with a chemotherapeutic agent.

In another embodiment of the invention, the compounds or compositions of the invention may be used in a method of preventing or treating cachexia in combination with a high calorie, high protein, high carbohydrate, vitamin B3, vitamin B12 and/or vitamin D supplement, antioxidant, omega fatty acid and/or polyphenol dietary intervention.

Sarcopenia and related disorders

Sarcopenia may be characterized by one or more of low muscle mass, low muscle strength, and low physical fitness.

An individual may be diagnosed for sarcopenia based on the AWGSOP (asian working group for geriatric sarcopenia) definition, for example as described by Chen et al 2014. Low muscle mass can generally be based on low extremity lean mass (ALM index) normalized to height squared, in particular ALM index less than 7.00kg/m2 for men and less than 5.40kg/m2 for women. Low physical performance may generally be based on walking speed, in particular walking speed less than 0.8 m/sec. Low muscle strength may generally be based on low grip strength, in particular less than 26kg for men and less than 18kg for women.

The diagnosis of sarcopenia in an individual may be based on the definition of EWGSOP (european working group for sarcopenia in elderly), e.g. as described by Crutz-Jentoft et al 2010. Low muscle mass can generally be based on low limb lean body mass (ALM index) normalized to height squared, in particular ALM index less than 7.23kg/m2 for men and less than 5.67kg/m2 for women. Low physical performance may generally be based on walking speed, in particular walking speed less than 0.8 m/sec. Low muscle strength may generally be based on low grip strength, in particular less than 30kg for men and less than 20kg for women.

The individual may be diagnosed with sarcopenia based on the definition of the national institute of health Foundation (FNIH), for example as described by Studenski et al 2014. Low muscle mass can generally be based on low extremity lean body mass (ALM) normalized to body mass index (BMI; Kg/m2), specifically ALM to BMI less than 0.789 for men and ALM to BMI less than 0.512 for women. Low physical performance may generally be based on walking speed, in particular walking speed less than 0.8 m/sec. Low muscle strength may generally be based on low grip strength, in particular less than 26kg for men and less than 16kg for women. Low muscle strength can also be generally based on low grip: body mass index, in particular the male grip: body mass index less than 1.00, and female grip: the body mass index is less than 0.56.

D3-creatine dilution is another method for measuring muscle mass. This approach is becoming more widely accepted as a robust standard and is expected to replace DXA in the future. The D3-creatine dilution method has been previously described in Clark et al, (1985) and Stimpson et al, (2013).

It is to be understood that the compounds, compositions and methods of the present invention may be beneficial in the prevention and/or treatment of sarcopenia and/or related conditions, in particular the maintenance or improvement of skeletal muscle mass and/or muscle function.

Myopathy and related disorders

Myopathy is a neuromuscular disorder, the primary symptom being muscle weakness due to dysfunction of muscle fibers. Other symptoms of myopathy may include muscle spasms, stiffness, and convulsions. Myopathies may be inherited (such as muscular dystrophy) or acquired (such as common muscle spasms).

Myopathies were grouped as follows: (i) congenital myopathy: it is characterized by a developmental delay in motor skills; skeletal and facial abnormalities are occasionally evident at birth; (ii) muscular dystrophy: it is characterized by gradual muscle weakness at will; sometimes evident at birth; (iii) mitochondrial myopathy: caused by genetic abnormalities of mitochondria (cellular structures that control energy); including Keams-Sayre syndrome, MELAS and MERRF muscle glycogen storage disease: caused by genetic mutations in enzymes that control the metabolism of glycogen and glucose (blood glucose); including Pompe, Andersen, and Cori diseases; (iv) myoglobinuria: caused by a disturbance in the metabolism of the fuel (myoglobin) necessary for the muscle to work; including McArdle's disease, Tarui's disease, and DiMauro's disease; (v) dermatomyositis: inflammatory myopathies of the skin and muscle; (vi) myositis ossificans: characterized in that bone grows in muscle tissue; (vii) familial periodic paralysis: it is characterized by the onset of weakness in the arms and legs; (viii) polymyositis, inclusion body myositis and related myopathies: skeletal myositis myopathy; (ix) neuromuscular rigidity: it is characterized by alternating episodes of twitching and stiffness; and stiff person syndrome: it is characterized by the onset of stiffness and reflex spasms (common muscle spasms and stiffness), and (x) tetany: it is characterized by prolonged twitching of the arms and legs. (reference: https:// www.ninds.nih.gov/disorders/all-disorders/myopathy-information-page).

It is to be understood that the compounds, compositions, and methods of the present invention may be beneficial in the prevention and/or treatment of the above-mentioned diseases or disorders, in particular in the maintenance or improvement of skeletal muscle mass and/or muscle function.

Muscular dystrophy

Muscular dystrophy is a group of genetic diseases characterized by progressive weakness and degeneration of skeletal or voluntary muscles that control movement. The main types of muscular dystrophy include: duchenne muscular dystrophy, Becker muscular dystrophy, limb girdle muscular dystrophy, facioscapulohumeral muscular dystrophy, congenital muscular dystrophy, oculopharyngeal muscular dystrophy, distal muscular dystrophy, Emery-Dreifuss muscular dystrophy, and myotonic dystrophy.

(reference: https:// www.medicalnewstoday.com/articles/187618.php)

It is to be understood that the compounds, compositions, and methods of the present invention may be beneficial in the prevention and/or treatment of the above-mentioned diseases or disorders, in particular in the maintenance or improvement of skeletal muscle mass and/or muscle function.

Post-operative and muscle trauma muscle injury recovery

Muscle damage can be caused by abrasion, stretching, or tearing, causing acute or chronic soft tissue damage to the muscles, tendons, or both. It can occur due to muscle fatigue, overuse, or misuse. It can occur after physical trauma such as a fall, break or overuse during physical activity. Muscle damage may also occur following surgical procedures such as arthroscopic joint replacement surgery.

It will be appreciated that the compounds, compositions and methods of the present invention may be beneficial in the prevention and/or treatment of the above-mentioned conditions of recovery following surgery and/or muscle trauma, in particular in the maintenance or improvement of skeletal muscle mass and/or muscle function.

Method of treatment

It will be appreciated that all references herein to treatment include curative, palliative and prophylactic treatment; in the context of the present invention, however, reference to prophylaxis is more often relevant to prophylactic treatment. Treatment may also include arresting the progression of the severity of the disease.

In one embodiment, the term "treatment" of any disease or disorder refers to ameliorating the disease or disorder (i.e., slowing or arresting or reducing the development of at least one of the clinical symptoms of the disease or its clinical symptoms). In another embodiment, "treating" refers to reducing or improving at least one physical parameter, including those that the patient may not recognize. In another embodiment, "treating" or "treatment" refers to modulating the disease or disorder, whether physical (e.g., stabilization of a recognizable symptom), physiological (e.g., stabilization of a physical parameter), or both. In another embodiment, "treating" or "treatment" refers to preventing or delaying the onset or development or progression of a disease or disorder. As used herein, an individual is "in need of treatment" if such individual would benefit biologically, medically or in quality of life from such treatment.

Individuals

The term "subject" refers to any animal, including humans and companion animals. Generally, the subject is a human or avian animal, bovine, canine, equine, feline, caprine, murine, ovine, and porcine animal. The subject may be a horse or a companion animal, such as a cat or dog. Preferably, the subject is a human.

Treatment of mammals, particularly humans, is preferred. However, both human and veterinary treatment are within the scope of the present invention.

For veterinary individuals, dog, cat, and equine individuals are preferred.

The invention may also be used with non-human animal subjects, such as: avian, bovine, ovine or porcine animals to optimize meat production by increasing skeletal muscle mass and/or function.

Muscle stem cells

As used herein, the term "muscle stem cell" may refer to a satellite cell, preferably a dormant and non-targeted satellite cell.

Satellite cells are precursors to skeletal muscle cells. In adult muscle, satellite cells are usually dormant, but can be activated and undergo myogenesis in response to disease or mechanical strain such as injury or exercise. Satellite cells are also involved in the normal growth of muscle. Upon activation, satellite cells may proliferate before undergoing myogenic differentiation to eventually fuse with existing muscle fibers or form new muscle fibers, depending on the size of the tissue wound. In addition to generating differentiated myogenic progeny, at least some satellite cells can self-renew, meeting the defined criteria for true stem cell retention. Pax7 is the best known and well characterized marker of muscle stem cell expression, i.e., muscle stem cells can be reliably identified based on their expression of the paired box transcription factor Pax 7. Muscle stem cells may also express NCAM, CD56, CD29, and/or CD82, i.e., muscle stem cells may be NCAM +, CD56+, CD29+, and/or CD82 +.

MyoD + is a directional marker that can be used to distinguish between dormant satellite cells and directional satellite cells.

Muscle function and muscle mass

The compounds, compositions, uses and methods disclosed herein can be used to maintain or increase muscle function and/or muscle mass.

The term "muscle function" refers to the ability of a muscle to function in a manner that does not negatively impact the life of an individual, and includes parameters of muscle strength, muscle contraction, muscle endurance, and/or muscle elasticity.

Suitable tests for assessing muscle function include: grip strength evaluation using a dynamometer; one repeat maximum for leg compressions, chest compressions, or leg extensions; walking speed; 6 minutes walk test; time up and go test (time up and go); simple physical ability test; fried debilitation criteria; and stair climb time assessment.

Muscle mass (which may be equivalent to muscle volume, muscle thickness, or muscle fiber size) may be measured by dual energy X-ray absorptiometry (DXA) or bioimpedance testing. Similarly, MRI can be used to assess muscle volume, and ultrasound can be used to assess muscle thickness and pinnate angle.

"muscle atrophy" may be a reduction in muscle mass, for example to a stage where muscle loss becomes weak. In one embodiment, the individual does not lose more than 10%, 5%, 4%, 3%, 2% or 1% of their muscle mass.

Preferably, the compounds, compositions, uses and methods disclosed herein are used to maintain or increase muscle mass.

The term "maintenance" means that a particular parameter, such as muscle function and/or muscle mass, remains substantially unchanged over a period of time (e.g., 5 years, 10 years, 15 years, 20 years, 25 years, 30 years, 40 years, 50 years, or more).

In one embodiment, muscle mass is increased by at least 1%, 2%, 3%, 4%, 5%, 10%, 15% or 20%.

In another embodiment, muscle mass is increased by 1% to 2.5%, 1% to 5%, 1% to 10%, or 1% to 20%.

Preferably, the muscle is skeletal muscle.

Examples

Example 1: selection of Compounds for modulating muscle Stem cells

Selection of human skeletal myoblasts

The present inventors developed a high-content screen to test compounds on human primary adult muscle cells in vitro. Human skeletal myoblasts (HSMM) were purchased from Lonza (R) ((R))https：//bioscience.lonza.com). These cells were isolated from the upper arm or leg muscle tissue of normal donors and used after the second passage. Several donors have been tested to ensure cell viability and purity prior to selecting the final donor for a 36 year old white female.

Muscle stem cell targeting assay

The primary screening assay is based on high level detection of two important myogenic regulators (Pax7 and MyoD) by immunofluorescence. Pax7 and MyoD are the primary markers of stem cell character and orientation of muscle stem cells and can be used to monitor muscle stem cell progeny. In particular, Pax7 marks early amplification, whereas MyoD is a myogenic late marker, and the combination of these markers defines different states of proliferation, differentiation and self-renewal.

Hit selection was mainly based on compounds that could enhance targeting to myogenic differentiation (Pax7-/MyoD + cells), which was particularly relevant in the context of cancer cachexia where defects in myogenic targeting have been shown to be a potential cause of muscle atrophy (He et al, 2013).

Human primary myoblasts were seeded at a density of 1' 000 cells per well in skeletal muscle growth medium (SKM-M, AMSbio) in 384-well plates. For treatment, compounds were added directly to myoblast cultures 16 hours after initial inoculation. All cultures were then grown for 96 hours. Cells were stained with antibodies to Pax7 and MyoD to determine Pax7 and MyoD expression, and counterstained with Hoechst 33342 to visualize the nuclei. MyoD + is defined as a cell that does not express Pax7 but expresses MyoD. Image acquisition was performed using the imagexpress (molecular devices) platform. Quantification was performed using a custom module analysis of multiwavelength cell scoring based on MetaXpress software. Figure 1 shows the number of Pax7+ cells and MyoD + cells normalized to the total number of cells in order to assess myogenic orientation.

Myogenesis assay

To confirm the results from the primary assay described above, a secondary assay of in vitro myogenesis was performed on all hits obtained in this primary screening assay. This secondary assay focuses on the later stages of myogenesis when muscle cells fuse together and form multinucleated myotubes. The assay is based on the detection of mature troponin-T protein expressed in the myotubes. Specifically, two measurements were performed:

fusion factor (which is the percentage of nuclei within the multinucleated myotube) was compared to total nuclei and gave a reading of muscle cell differentiation.

Myotube area (measured based on troponin T staining) and gives readings on myotube size and myofiber growth.

Human primary myoblasts were seeded at a density of 3' 000 cells per well in skeletal muscle growth medium (SKM-M, AMSbio) in 384-well plates. After one day, differentiation was induced by medium exchange. For treatment, compounds were added directly to myoblast cultures for 96 hours. Myotubes were stained with antibodies against troponin T to determine troponin T expression and counterstained with Hoechst 33342 to visualize the nuclei. Image acquisition was performed using the imagexpress (molecular devices) platform. Quantification was performed using a custom module analysis of multiwavelength cell scoring based on MetaXpress software. Figure 2 shows cell viability assessed by cell number, myogenic differentiation measured by fusion factors (expressed as the percentage of nuclei within troponin T-positive myotubes), and myofiber growth (expressed as the area covered by troponin T-positive myotubes).

Example 2: clinical trials with compound GW5074

For phase I/IIa clinical studies using GW5074, subjects received a dosing regimen of GW5074 and sorafenib (within the safe dosing range defined in phase I) combination therapy for > 168 days (24 weeks). Dosage of GW 5074: 750mg to 1500mg (daily dose).

https：//clinicaltrials.gov/ct2/show/NCT03406364

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