Composition for senescent cell removal

Abstract

A compound expressed by R1-S-S-S-R2 [R1 and R2 are each independently a group in which a sulfhydryl group has been excluded from an optionally-protected pantetheine, a group in which a sulfhydryl group has been excluded from an optionally-protected glutathione, or a group in which a sulfhydryl group has been excluded from an optionally-protected cysteine], or a pharmaceutically-acceptable salt thereof or a compound expressed by formula (VI), a chemically-acceptable salt thereof, or cyclodextrin inclusion complexes of the above are able to selectively kill senescent cells. In formula (VI), X is hydroxyl group or the like.

Description

Composition for removing senescent cells 【0001】 This disclosure relates to a composition for removing senescent cells. 【0002】 Normal human somatic cells have a limited number of divisions they can undergo. Cellular senescence refers to the changes in cells that progress towards this division limit. Senescent cells are defined as cells whose cell proliferation or cell cycle has irreversibly stopped. Senescent cells escape cell death and accumulate as an individual ages, and are recognized as the main cause of age-related decline in tissue function or loss of regenerative capacity. 【0003】 Studies using progeria model mice have reported that artificially removing senescent cells from aged individuals significantly delays the onset of age-related diseases such as arteriosclerosis and renal impairment, and even extends lifespan (Non-Patent Literature 1). The development of drugs that selectively kill senescent cells present in the body (senolytic drugs) is expected to lead to an extension of healthy lifespan and new prevention and / or treatment of age-related diseases. 【0004】 As a means of selectively killing senescent cells, for example, selective cell death induction of senescent cells by inhibiting the glutamine metabolic pathway (glutaminolysis), particularly glutaminase 1 (GLS1), has been discovered (Patent Document 1). This is based on the fact that in senescent cells, the conversion reaction from citrate to isocitrate is inhibited due to an increase in reactive oxygen species, and the production of α-ketoglutarate and the subsequent rotation of the citric acid cycle depend on the glutamine metabolic pathway. 【0005】 International Publication No. 2020 / 095971 【0006】 Baker et al., "Clearance of p16Ink4a-positive senescent cells delays aging-associated disorders", Nature, 2011, 479(7372): 232-236 【0007】 The object of the present invention is to provide a compound that selectively kills senescent cells. 【0008】The inventors of the present invention have found that a predetermined trisulfide compound selectively kills senescent cells and suppresses the expression of GLS1 in senescent cells, and thus have completed the present invention. 【0009】 That is, the present disclosure relates to the following [1] to

[10] . [1] A compound represented by R １ -S-S-S-R ２ where R １ and R ２ are each independently a group obtained by removing a sulfhydryl group from optionally protected pantethine, a group obtained by removing a sulfhydryl group from optionally protected glutathione, or a group obtained by removing a sulfhydryl group from optionally protected cysteine], or a pharmaceutically acceptable salt thereof or a compound represented by formula (VI). [In formula (VI), X represents -OR ６１ or -NR ６２ R ６３ ; R ６１ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms; R ６２ and R ６３ each independently represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and the alkyl group may have one or more substituents selected from the group consisting of an amino group and a carboxyl group], a pharmaceutically acceptable salt thereof or a cyclodextrin inclusion complex thereof, a composition for removing senescent cells. [2] A compound represented by R １ -S-S-S-R ２ is a compound represented by formula (I). [R １１ and R １２ are each independently a hydrogen atom, an alkyl group having 1 to 9 carbon atoms or an acyl group having 1 to 5 carbon atoms, or R １１ and R １２ together may form a methylene group or a carbonyl group optionally having an alkyl group having 1 to 4 carbon atoms; R １３ and R １４ are each independently a hydrogen atom, an alkyl group having 1 to 9 carbon atoms or an acyl group having 1 to 5 carbon atoms, or R １３ and R １４[Together, these are a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms], a compound represented by formula (II). [In formula (II), R ２１ and R ２２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R ２１ and R ２２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R ２３ and R ２４ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ２５ Compounds represented by formula (III), where is a protecting group for a hydrogen atom or an amino group. [In formula (III), R ３１ and R ３２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R ３１ and R ３２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R ３３ R is a protecting group for a hydrogen atom or a carboxyl group. ３４ Compounds represented by formula (IV), where is a protecting group for a hydrogen atom or an amino group. [In formula (IV), R ４１ , R ４２ , R ４３ and R ４４ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ４５ and R ４６ [Each is independently a protecting group for a hydrogen atom or an amino group] or a compound represented by formula (V) [In formula (V), R ５１ and R ５２ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ５３ and R ５４ The composition according to [1], wherein each of the following is independently a protecting group for a hydrogen atom or an amino group. [3]R １ -S-S-S-R ２A compound represented by or a compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa). The composition according to [1]. [4] The senescent cells are cells that highly express glutaminase 1, the composition according to any one of [1] to [3]. [5] The senescent cells are fibroblasts, the composition according to any one of [1] to [4]. [6] R １ -S-S-S-R ２ A pharmaceutical composition for the treatment or prevention of age-related diseases, comprising a compound represented by formula (VI), a pharmaceutically acceptable salt thereof, or a compound represented by formula (VI), a pharmaceutically acceptable salt thereof, or a cyclodextrin inclusion complex thereof. [7]R １ -S-S-S-R ２ The pharmaceutical composition according to [6], wherein the compound represented is the compound represented by formula (I), the compound represented by formula (II), the compound represented by formula (III), the compound represented by formula (IV), or the compound represented by formula (V). [8]R １ -S-S-S-R ２The pharmaceutical composition according to [6], wherein the compound represented by or the compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa). [9] The pharmaceutical composition according to any one of [6] to [8], wherein the age-related disease is arteriosclerosis, arteriosclerotic cerebrovascular disease, osteoporosis, cataract, glaucoma, presbyopia, age-related macular degeneration, cerebral infarction, dementia, Parkinson's disease, cerebral hemorrhage, pulmonary fibrosis, chronic obstructive pulmonary disease, emphysema, type 2 diabetes, chronic renal failure, chronic kidney disease, cardiac hypertrophy, heart failure, hypertension, cirrhosis, fatty liver, non-alcoholic steatohepatitis, sarcopenia, skeletal muscle atrophy, emaciation, dyslipidemia, osteoarthritis, age-related lower back pain, age-related joint pain, frailty, age-related alopecia, age-related hearing loss, seborrheic dermatitis, pruritus, Hutchinson-Gilford progeria syndrome, Werner syndrome, Cockayne syndrome, or Rothmond-Thomson syndrome. 【0010】 This disclosure also relates to the following

[10] to

[45] .

[10] R １ -S-S-S-R ２ A senescent cell remover containing a compound represented by formula (VI), a pharmaceutically acceptable salt thereof, or a compound represented by formula (VI), a pharmaceutically acceptable salt thereof, or a cyclodextrin inclusion complex thereof.

[11] R １ -S-S-S-R ２ The senescent cell remover according to

[10] , wherein the compound represented is the compound represented by formula (I), the compound represented by formula (II), the compound represented by formula (III), the compound represented by formula (IV), or the compound represented by formula (V).

[12] R １ -S-S-S-R ２ The senescent cell scavenger according to

[10] , wherein the compound represented by or the compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa).

[13] The senescent cell scavenger according to any one of

[10] to

[12] , wherein the senescent cells are cells that highly express glutaminase 1.

[14] The senescent cell scavenger according to any one of

[10] to

[13] , wherein the senescent cells are fibroblasts.

[15] R １-S-S-S-R ２ A therapeutic or prophylactic agent for age-related diseases comprising a compound represented by formula (VI), or a pharmaceutically acceptable salt thereof, or a compound represented by formula (VI), a pharmaceutically acceptable salt thereof, or a cyclodextrin inclusion complex thereof.

[16] R １ -S-S-S-R ２ The therapeutic or prophylactic agent according to

[15] , wherein the compound represented is the compound represented by formula (I), the compound represented by formula (II), the compound represented by formula (III), the compound represented by formula (IV), or the compound represented by formula (V).

[17] R １ -S-S-S-R ２ The therapeutic or prophylactic agent according to

[15] , wherein the compound represented by or the compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa).

[18] The age-related disease is arteriosclerosis, arteriosclerotic cerebrovascular disease, osteoporosis, cataract, glaucoma, presbyopia, age-related macular degeneration, cerebral infarction, dementia, Parkinson's disease, cerebral hemorrhage, pulmonary fibrosis, chronic obstructive pulmonary disease, emphysema, type 2 diabetes, chronic renal failure, chronic kidney disease, cardiac hypertrophy, heart failure, hypertension, cirrhosis, fatty liver, non-alcoholic steatohepatitis, sarcopenia, skeletal muscle atrophy, emaciation, dyslipidemia, osteoarthritis, age-related lower back pain, age-related joint pain, frailty, age-related alopecia, age-related hearing loss, seborrheic dermatitis, pruritus, Hutchinson-Gilford progeria syndrome, Werner syndrome, Cockayne syndrome, or Rothmond-Thomson syndrome, as described in any of

[15] to

[17] .

[19] R １ -S-S-S-R ２ A method for removing senescent cells from a patient, comprising administering to the patient a compound represented by formula (VI), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable salt thereof, or a cyclodextrin inclusion complex thereof.

[20] R １ -S-S-S-R ２ The method according to

[19] , wherein the compound represented is the compound represented by formula (I), the compound represented by formula (II), the compound represented by formula (III), the compound represented by formula (IV), or the compound represented by formula (V).

[21] R１ -S-S-S-R ２ The method according to

[19] , wherein the compound represented by or the compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa).

[22] The method according to any one of

[19] to

[21] , wherein the senescent cells are cells that highly express glutaminase 1.

[23] The method according to any one of

[19] to

[22] , wherein the senescent cells are fibroblasts.

[24] R １ -S-S-S-R ２ A method for treating or preventing age-related diseases, comprising administering to a patient a compound represented by formula (VI), a pharmaceutically acceptable salt thereof, or a cyclodextrin inclusion complex thereof.

[25] R １ -S-S-S-R ２ The method according to

[24] , wherein the compound represented is the compound represented by formula (I), the compound represented by formula (II), the compound represented by formula (III), the compound represented by formula (IV), or the compound represented by formula (V).

[26] R １ -S-S-S-R ２ The method according to

[24] , wherein the compound represented by or the compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa).

[27] The method according to any one of

[24] to

[26] , wherein the age-related disease is arteriosclerosis, arteriosclerotic cerebrovascular disease, osteoporosis, cataract, glaucoma, presbyopia, age-related macular degeneration, cerebral infarction, dementia, Parkinson's disease, cerebral hemorrhage, pulmonary fibrosis, chronic obstructive pulmonary disease, emphysema, type 2 diabetes, chronic renal failure, chronic kidney disease, cardiac hypertrophy, heart failure, hypertension, cirrhosis, fatty liver, non-alcoholic steatohepatitis, sarcopenia, skeletal muscle atrophy, emaciation, dyslipidemia, osteoarthritis, age-related lower back pain, age-related joint pain, frailty, age-related alopecia, age-related hearing loss, seborrheic dermatitis, pruritus, Hutchinson-Gilford progeria syndrome, Werner syndrome, Cockayne syndrome, or Rothmond-Thomson syndrome.

[28] R for use in a method for removing senescent cells １-S-S-S-R ２ Compounds represented by formula (VI), or pharmaceutically acceptable salts thereof, or compounds represented by formula (VI), pharmaceutically acceptable salts thereof, or cyclodextrin inclusion complexes thereof.

[29] R １ -S-S-S-R ２ The compound, salt, or inclusion body according to

[28] , wherein the compound represented is the compound represented by formula (I), the compound represented by formula (II), the compound represented by formula (III), the compound represented by formula (IV), or the compound represented by formula (V).

[30] R １ -S-S-S-R ２ The compound, salt, or inclusion body according to

[28] , wherein the compound represented by or the compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa).

[31] The compound, salt, or inclusion body according to any one of

[28] to

[30] , wherein the senescent cell is a cell that highly expresses glutaminase 1.

[32] The compound, salt, or inclusion body according to any one of

[28] to

[31] , wherein the senescent cell is a fibroblast.

[33] For use in methods of treating or preventing age-related diseases, R １ -S-S-S-R ２ Compounds represented by formula (VI), or pharmaceutically acceptable salts thereof, or compounds represented by formula (VI), pharmaceutically acceptable salts thereof, or cyclodextrin inclusion complexes thereof.

[34] R １ -S-S-S-R ２ The compound, salt, or inclusion body according to

[33] , wherein the compound represented is the compound represented by formula (I), the compound represented by formula (II), the compound represented by formula (III), the compound represented by formula (IV), or the compound represented by formula (V).

[35] R １ -S-S-S-R ２The compound represented by or the compound represented by formula (VI) is the compound, salt or inclusion complex described in

[33] , which is represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb) or formula (VIa).

[36] The age-related disease is arteriosclerosis, atherosclerotic cerebrovascular disease, osteoporosis, cataract, glaucoma, presbyopia, age-related macular degeneration, cerebral infarction, dementia, Parkinson's disease, cerebral hemorrhage, pulmonary fibrosis, chronic obstructive pulmonary disease, emphysema, type 2 diabetes, chronic renal failure, chronic kidney disease, cardiac hypertrophy, heart failure, hypertension, liver cirrhosis, fatty liver, non-alcoholic steatohepatitis, sarcopenia, skeletal muscle atrophy, emaciation, dyslipidemia, osteoarthritis, low back pain associated with aging, joint pain associated with aging, frailty, age-related hair loss, age-related hearing loss, seborrheic dermatitis, pruritus, Hutchinson-Gilford progeria syndrome, Werner syndrome, Cockayne syndrome or Rothmund-Thomson syndrome, the compound, salt or inclusion complex described in any of

[33] to

[35] .

[37] For the production of a composition for removing senescent cells, R １ -S-S-S-R ２ Use of the compound represented by , or a pharmaceutically acceptable salt thereof or the compound represented by formula (VI), a pharmaceutically acceptable salt thereof or a cyclodextrin inclusion complex thereof.

[38] R １ -S-S-S-R ２ The compound represented by is the compound represented by formula (I), the compound represented by formula (II), the compound represented by formula (III), the compound represented by formula (IV) or the compound represented by formula (V), the use described in

[37] .

[39] R １ -S-S-S-R ２ The compound represented by or the compound represented by formula (VI) is the compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb) or formula (VIa), the use described in

[37] .

[40] The senescent cells are cells that highly express glutaminase 1, the use described in any of

[37] to

[39] .

[41] The senescent cells are fibroblasts, the use described in any of

[37] to

[40] .

[42] For the production of a pharmaceutical composition for the treatment or prevention of age-related diseases, R １ -S-S-S-R ２Use of a compound represented by , or a pharmaceutically acceptable salt thereof, or a compound represented by formula (VI), a pharmaceutically acceptable salt thereof, or a cyclodextrin inclusion complex thereof.

[43] R １ -S-S-S-R ２ The use according to

[42] , wherein the compound represented by is a compound represented by formula (I), a compound represented by formula (II), a compound represented by formula (III), a compound represented by formula (IV), or a compound represented by formula (V).

[44] R １ -S-S-S-R ２ The use according to

[42] , wherein the compound represented by or the compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa).

[45] The age-related disease is arteriosclerosis, atherosclerotic cerebrovascular disease, osteoporosis, cataract, glaucoma, presbyopia, age-related macular degeneration, cerebral infarction, dementia, Parkinson's disease, cerebral hemorrhage, pulmonary fibrosis, chronic obstructive pulmonary disease, emphysema, type 2 diabetes, chronic renal failure, chronic kidney disease, cardiac hypertrophy, heart failure, hypertension, liver cirrhosis, fatty liver, non-alcoholic steatohepatitis, sarcopenia, skeletal muscle atrophy, emaciation, dyslipidemia, osteoarthritis, low back pain associated with aging, joint pain associated with aging, frailty, age-related hair loss, age-related hearing loss, seborrheic eczema, pruritus, Hutchinson-Gilford progeria syndrome, Werner syndrome, Cockayne syndrome, or Rothmund-Thomson syndrome, the use according to any one of

[42] to

[44] . 【0011】This disclosure provides a compound that selectively kills senescent cells. Such a compound is thought to selectively kill senescent cells by suppressing GLS1 expression in those cells. Therefore, it may be possible to treat diseases in which GLS1 expression is reported to be upregulated and cellular senescence is suspected to be involved. Examples of such diseases include arteriosclerosis, arteriosclerotic cerebrovascular and cardiovascular diseases, osteoporosis, cataracts, glaucoma, presbyopia, age-related macular degeneration, cerebral infarction, dementia, Parkinson's disease, cerebral hemorrhage, pulmonary fibrosis, chronic obstructive pulmonary disease, emphysema, type 2 diabetes, chronic renal failure, chronic kidney disease, cardiac hypertrophy, heart failure, hypertension, cirrhosis, fatty liver, non-alcoholic steatohepatitis, sarcopenia, skeletal muscle atrophy, emaciation, dyslipidemia, osteoarthritis, age-related lower back pain, age-related joint pain, frailty, age-related alopecia, age-related hearing loss, seborrheic dermatitis, pruritus, Hutchinson-Gilford progeria syndrome, Werner syndrome, Cockayne syndrome, and Rothmond-Thomson syndrome. 【0012】 This graph shows the effect of different cell division counts (PDLs) of normal human dermal fibroblasts (NB1RGB) on IL-1α expression levels. This graph shows the effect of trisulfide compounds on the cell viability of NB1RGB cells (PDL ≥ 50). This graph shows the effect of pantetheine trisulfide and pantethine on the cell viability of NB1RGB cells (PDL = 12). This graph shows the effect of pantetheine trisulfide and pantethine on the cell viability of NB1RGB cells (PDL ≥ 50). This graph shows the effect of trisulfide compounds on GLS1 expression in NB1RGB cells (PDL ≥ 50). GLS1 expression levels are expressed as relative values ​​with the control set to 1. 【0013】 A composition for removing senescent cells according to one embodiment of the present disclosure, R １ -S-S-S-R ２ Compound represented by [R １ and R ２Each of these is independently a group obtained by removing a sulfhydryl group from protected pantetheine, a group obtained by removing a sulfhydryl group from protected glutathione, or a group obtained by removing a sulfhydryl group from protected cysteine, or a pharmaceutically acceptable salt thereof or a compound represented by formula (VI). [In equation (VI), X is -OR ６１ or -NR ６２ R ６３ R ６１ R represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms. ６２ and R ６３ Each of these independently represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and the alkyl group may have one or more substituents selected from the group consisting of an amino group and a carboxyl group, and the mixture contains a pharmaceutically acceptable salt thereof or a cyclodextrin inclusion complex thereof. 【0014】 R １ and R ２ In this context, "protected" means that the hydroxyl, carboxyl, and amino groups contained in pantetheine, glutathione, and cysteine ​​are protected by their respective protecting groups. Such protecting groups are described in Wuts, "Greene's Protective Groups in Organic Synthesis", John Wiley & Sons Inc, 2014. 【0015】 In equation (VI), R ６１ This may be, for example, a hydrogen atom, a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, or a hexyl group, preferably R ６１ R is a hydrogen atom. ６２ and R ６３ Each of these may independently be a hydrogen atom, a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, or other alkyl groups. Furthermore, these alkyl groups may have substituents on one or both of an amino group and a carboxyl group. ６２ and R ６３R may be, for example, the group represented by formula (6) (where * indicates a bond). ６２ and R ６３ Both may be hydrogen atoms, R ６２ is a hydrogen atom and R ６３ The base may also be represented by formula (6). 【0016】 In the cyclodextrin inclusion complex of the compound represented by formula (VI) or a pharmaceutically acceptable salt thereof, the cyclodextrin may be α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, or a derivative thereof. Here, "derivative" means that at least one hydrogen atom of the hydroxyl group of each cyclodextrin is substituted with an alkyl group or sugar which may have substituents. Examples of cyclodextrin derivatives include methyl-α-cyclodextrin, methyl-β-cyclodextrin, methyl-γ-cyclodextrin, dimethyl-α-cyclodextrin, dimethyl-β-cyclodextrin, dimethyl-γ-cyclodextrin, hydroxyethyl-α-cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxyethyl-γ-cyclodextrin, 2-hydroxypropyl-α-cyclodextrin, and 2-hydroxypropyl-β-cyclodextrin. Rodextrin, 2-hydroxypropyl-γ-cyclodextrin, glucosyl-α-cyclodextrin, glucosyl-β-cyclodextrin, glucosyl-γ-cyclodextrin, maltosyl-α-cyclodextrin, maltosyl-β-cyclodextrin, maltosyl-γ-cyclodextrin, sulfobutyl ether-α-cyclodextrin, sulfobutyl ether-β-cyclodextrin, sulfobutyl ether-γ-cyclodextrin, etc. can be used. 【0017】 The compound represented by formula (VI) can be produced, for example, by the method described in WO2022 / 045212. 【0018】 R １ -S-S-S-R ２The compound represented by may be any compound represented by the following formulas (I) to (V). In this disclosure, the compound represented by formula (I) may also be referred to as "compound (I)," and compounds represented by other chemical formulas may be referred to similarly. [R １１ and R １２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R １１ and R １２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R １３ and R １４ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R １３ and R １４ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms. [In formula (II), R ２１ and R ２２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R ２１ and R ２２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R ２３ and R ２４ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ２５ [It is a protecting group for a hydrogen atom or an amino group.] [In formula (III), R ３１ and R ３２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R ３１ and R ３２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R ３３ R is a protecting group for a hydrogen atom or a carboxyl group. ３４ [It is a protecting group for a hydrogen atom or an amino group.] [In formula (IV), R ４１ , R ４２, R ４３ and R ４４ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ４５ and R ４６ Each of these is independently a protecting group for a hydrogen atom or an amino group. [In formula (V), R ５１ and R ５２ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ５３ and R ５４ Each of these is independently a protecting group for a hydrogen atom or an amino group. 【0019】 In formulas (I) to (III), examples of C1-C4 alkyl groups include methyl, ethyl, n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and cyclobutyl groups. In formula (I), examples of C1-C5 acyl groups include formyl (methanoyl), acetyl (ethanolyl), n-propanoyl, isopropanoyl, cyclopropanoyl, n-butanoyl, isobutanoyl, sec-butanoyl, tert-butanoyl, and cyclobutanoyl groups. From the viewpoint of lowering the molecular weight of the compound, the C1-C4 alkyl group is preferably a methyl or ethyl group, and the C1-C5 acyl group is preferably a formyl or acetyl group. 【0020】 In formulas (I) to (III), R １１ and R １２ , R １３ and R １４ , R ２１ and R ２２ , R ３１ and R ３２ One example of a combination is (R １１ , R １２ ), (R １３ , R １４ ), (R ２１ , R ２２ ), (R ３１ , R ３２Examples of combinations include (hydrogen atom, hydrogen atom), (hydrogen atom, methyl group), (methyl group, hydrogen atom), (methyl group, methyl group), (hydrogen atom, formyl group), (formyl group, hydrogen atom), and (formyl group, formyl group). 【0021】 R １１ and R １２ , R １３ and R １４ , R ２１ and R ２２ or R ３１ and R ３２ When these combine to form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, R １ and R ２ Examples of combinations include a methylene group, a methylene group having one methyl group, a methylene group having two methyl groups, and a carbonyl group. 【0022】 In formulas (II) to (V), the "carboxyl protecting group" can refer to carboxylic acid protecting groups that are generally known to those skilled in the art, such as the methyl group, ethyl group, isopropyl group, trialkylsilyl group, tert-butyl group, and benzyl group. 【0023】 In formulas (II) to (V), the "amino group protecting group" can refer to amino group protecting groups that are generally known to those skilled in the art, such as amide protecting groups such as formyl group, acetyl group, benzoyl group, nicotinoyl group, trichloroacetyl group, and trifluoroacetyl group; cyclic imide protecting groups such as phthaloyl group and 2,3-diphenylmaleoyl group; sulfonamide protecting groups such as p-toluenesulfonyl group; and carbamate protecting groups such as tert-butyloxycarbonyl group, methyloxycarbonyl group, ethyloxycarbonyl group, benzyloxycarbonyl group, allyloxycarbonyl group, p-methoxybenzylcarbonyl group, p-nitrobenzyloxycarbonyl group, and 9-fluorenylmethyloxycarbonyl group. 【0024】 R １ -S-S-S-R ２The compound represented by or the compound represented by formula (VI) may be a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa). 【0025】 The compound represented by formula (Ia) is, in formula (I), R １１ , R １２ , R １３ and R １４ It is a pantetheine trisulfide (Ptt-SSS-Ptt; Ptt represents the group obtained by removing the sulfhydryl group from pantetheine; sometimes written as PTN-SSS) in which all are hydrogen atoms. The compound represented by formula (Ib) is in formula (I) where R １１ and R １２ Together they form a carbonyl group, R １３ and R １４ Together they form a carbonyl group, and the compound is a pantetheine trisulfide diacetal (Ptt-SSS-Ptt-diacetal). The compound represented by formula (IIa) is, in formula (II), R ２１ , R ２２ , R ２３ , R ２４ and R ２５ Pantetheine glutathione trisulfide (Ptt-SSS-G; G represents the group obtained by removing the sulfhydryl group from glutathione) is a compound in which all are hydrogen atoms. The compound represented by formula (IIIa) is in formula (III) where R ３１ , R ３２ , R ３３ and R ３４ The compound represented by formula (IVa) is pantetheine cysteine ​​trisulfide (Ptt-SSS-C; C represents the group obtained by removing the sulfhydryl group from cysteine). ４１ , R ４２ , R ４３ , R ４４ , R ４５and R ４６ It is glutathione trisulfide (G-SSS-G), where all atoms are hydrogen atoms. The compound represented by formula (Va) is, in compound (V), R ５１ , R ５２ , R ５３ and R ５４ It is a cysteine ​​trisulfide (C-SSS-C) where all atoms are hydrogen atoms. The compound represented by formula (Vb) is a compound in which, in compound (V), R ５１ and R ５２ is a hydrogen atom, R ５３ and R ５４ The compound represented by formula (VIa) is an acetyl group, and is an N-acetylcysteine ​​trisulfide (NAC-SSS-NAC; NAC represents the group obtained by removing the sulfhydryl group from N-acetylcysteine). The compound represented by formula (VIa) is an α-lipoic acid trisulfide (LA-SSS) in compound (VI), where X is a hydroxyl group. 【0026】 The compound represented by formula (I) can be produced, for example, by the method described in WO2024 / 143400. 【0027】 The compounds represented by formulas (II) and (III) can be produced, for example, by the following method. In the above reaction scheme, R Ａ teeth or Represents R Ｂ teeth or This represents X １ represents hydrogen, 9-methylfluorene, etc., X ２ The symbols represent halogens, alkoxy groups, etc. 【0028】 Examples of reaction solvents for step A include water, aqueous sulfuric acid solution, aqueous ethanol solution, and aqueous acetonitrile solution, and the amount of solvent is R Ａ -SS-R ＡThe amount can be 1 mL to 500 mL per 1 g. Examples of oxidizing agents that can be used include potassium peroxymonosulfate (sold under trade names such as Oxone®), peracetic acid, hydrogen peroxide (may be used together with a catalytic amount of methyltrioxorenium), and sodium periodate, and the amount of oxidizing agent used is R Ａ -SS-R Ａ The amount can be 0.8 to 4.0 equivalents per 1 equivalent. Examples of sulfur sources that can be used include sodium sulfide (sodium sulfide notahydrate), potassium sulfide, sodium hydrogen sulfide, potassium hydrogen sulfide, and hydrogen sulfide, and the amount of sulfur source used is R Ａ -SS-R Ａ For every 1 equivalent, the amount can range from 0.5 to 4.0 equivalents. Ｂ -SS-R Ｂ The amount is R Ａ -SS-R Ａ The amount of 0.8 to 2.0 equivalents can be used per 1 equivalent. The reaction temperature can be -20°C to 30°C. The reaction time can be 15 minutes to 2 days. 【0029】 Examples of reaction solvents for step B include water and isopropanol, and the amount of solvent is R Ａ - The amount can be 1 mL to 500 mL per 1 g of SH. Examples of condensing agents used include N,N'-Thiodymethalamide, and the amount of condensing agent used is R Ａ - The amount can be 0.8 to 2.0 equivalents relative to SH1 equivalent. Ｂ - The amount of SH is R Ａ -The amount can be 0.8 to 2.0 equivalents per SH1 equivalent. The reaction temperature can be -20°C to 30°C. The reaction time can be 15 minutes to 2 days. 【0030】 Examples of reaction solvents for step C include water, aqueous sulfuric acid solution, aqueous ethanol solution, and aqueous acetonitrile solution, and the amount of solvent is R Ａ -SS-R ＡThe amount can be 1 mL to 500 mL per 1 g. Examples of oxidizing agents that can be used include potassium peroxymonosulfate, peracetic acid, hydrogen peroxide (which may be used together with a catalytic amount of methyltrioxorenium), and sodium periodate, and the amount of oxidizing agent used is R Ａ -SS-R Ａ The amount can be 0.8 to 2.0 equivalents per 1 equivalent. Examples of sulfur sources that can be used include glutathione and cysteine, and the amount of sulfur source used is R Ａ -SS-R Ａ The amount of 0.8 to 2.0 equivalents can be used per 1 equivalent. The reaction temperature can be -20°C to 30°C. The reaction time can be 15 minutes to 2 days. 【0031】 Examples of reaction solvents for step D include aprotic solvents (dichloromethane, tetrahydrofuran, etc.) and protic solvents (methanol, isopropanol, etc.), and the amount of solvent is R Ａ -SS-X １ or R Ａ -S-X １ The amount can be 1 mL to 500 mL per 1 g. Examples of reagents used include sodium methoxide, diazabicycloundecene (DBU), and tris(pentafluorophenyl)borane, and the amount of reagent used is R Ａ -SS-X １ or R Ａ -S-X １ For every 1 equivalent, the amount can range from 0.5 to 4.0 equivalents. Ｂ -S-X ２ or R Ｂ -SS-X ２ The amount is R Ａ -SS-X １ or R Ａ -S-X １ The amount of 0.8 to 2.0 equivalents can be used per 1 equivalent. The reaction temperature can be -20°C to 30°C. The reaction time can be 15 minutes to 2 days. 【0032】The compounds represented by formulas (IV) and (V) can be produced, for example, by the method described in WO2021 / 200487. 【0033】 R １ -S-S-S-R ２ Examples of pharmaceutically acceptable salts of compounds represented by formula (VI) include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; salts with organic acids such as acetic acid, succinic acid, fumaric acid, maleic acid, tartaric acid, citric acid, lactic acid, stearic acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, and p-toluenesulfonic acid; salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as calcium and magnesium; ammonium salts; and salts with amino acids such as arginine. 【0034】 Composition for removing senescent cells, R １ -S-S-S-R ２ In addition to compounds represented by formula (VI), or their pharmaceutically acceptable salts, or compounds represented by formula (VI), their pharmaceutically acceptable salts, or cyclodextrin inclusion complexes thereof, the product may contain pharmaceutically acceptable additives. Examples of additives include stabilizers such as sugars (sucrose, trehalose, maltose, lactose, etc.), sugar alcohols (sorbitol, etc.), amino acids (L-arginine, etc.), water-soluble polymers (HES (hydroxyethyl starch), PVP (polyvinylpyrrolidone), etc.), nonionic surfactants (polysorbate, poloxamer, etc.), pH adjusters such as sodium phosphate buffer and histidine buffer, isotonic agents such as sodium chloride, and excipients such as mannitol, glycine, sodium chloride, and sucrose. 【0035】Compositions for removing senescent cells can be formulated, for example, as parenteral or oral preparations. Parenteral preparations include, for example, injections, nasal drops, eye drops, sublingual preparations, transdermal preparations (topical applications, patches, etc.), pulmonary preparations (inhalants), transnasal preparations (inhalants), enteral preparations (suppositories), or transvaginal preparations (vaginal suppositories). Injectable preparations include, for example, subcutaneous injections, intramuscular injections, intravenous injections, intraperitoneal injections, intra-articular preparations, or intraocular preparations. Oral preparations can be formulated, for example, as tablets, granules, fine granules, powders, or capsules. 【0036】Since senescent cells highly express glutaminase 1 (GLS1), the senescent cell removal composition can efficiently remove cells that highly express glutaminase 1. High glutaminase 1 expression means that increased expression of glutaminase 1 is essential for cell survival. For example, cells may increase glutaminase 1 expression to increase ammonia production in order to cope with the shift in intracellular pH towards the acidic side due to lactic acid produced by increased glycolysis. Also, if cells depend on glutamate for energy production necessary for survival, they may increase glutaminase 1 expression to obtain glutamate, which is the raw material for survival. Furthermore, the senescent cell removal composition is effective against senescent cells present in various tissues and organs, regardless of the type of senescent cell. For example, senescent cells are a factor that impairs the function of various tissues such as the skin, liver, kidneys, lungs, heart, and brain. Senescent cells include, for example, mammary epithelial cells, keratinocytes, cardiomyocytes, chondrocytes, endothelial cells (large vessels), endothelial cells (microvessels), epithelial cells, fibroblasts, dermal papilla cells, hepatocytes, melanocytes, osteoblasts, adipocytes, immune system cells, skeletal muscle cells, smooth muscle cells, adipocytes, neurons, glial cells, contractile cells, exocrine epithelial cells, extracellular matrix cells, hormone-secreting cells, keratinized epithelial cells, pancreatic islet cells, lens cells, mesenchymal stem cells, pancreatic adenocarcinoma cells, Panethoocytes of the small intestine, hematopoietic system cells, nervous system cells, cells supporting sensory organs and peripheral nerve cells, and moist stratified barrier epithelial cells, preferably fibroblasts. The senescent cell removal composition targets various senescent cells and is expected to promote tissue rejuvenation and / or functional recovery through their removal. 【0037】Compositions for removing senescent cells can also be used as pharmaceutical compositions for the treatment or prevention of age-related diseases. Age-related diseases are diseases whose frequency increases as aging progresses, and include, for example, neurodegenerative diseases for which adequate treatments have not been established, such as Alzheimer's disease and Parkinson's disease; cardiovascular diseases, which are the main cause of death in middle-aged and elderly people; chronic renal failure, which affects the quality of life (QOL) of patients and also places a significant medical, economic, and social burden on them; retinal degenerative diseases; hearing impairment; infectious diseases and their sequelae; and cancer. More specific age-related diseases include arteriosclerosis, arteriosclerotic cerebrovascular and cardiovascular diseases, osteoporosis, cataracts, glaucoma, presbyopia, age-related macular degeneration, cerebral infarction, dementia, Parkinson's disease, cerebral hemorrhage, pulmonary fibrosis, chronic obstructive pulmonary disease, emphysema, type 2 diabetes, chronic renal failure, chronic kidney disease, cardiac hypertrophy, heart failure, hypertension, cirrhosis, fatty liver, non-alcoholic steatohepatitis, sarcopenia, skeletal muscle atrophy, emaciation, dyslipidemia, osteoarthritis, age-related lower back pain, age-related joint pain, frailty, age-related alopecia, age-related hearing loss, seborrheic dermatitis, pruritus, Hutchinson-Gilford progeria syndrome, Werner syndrome, Cockayne syndrome, and Rothmond-Thomson syndrome. 【0038】 Treatment of age-related diseases means suppressing or alleviating the various symptoms of age-related diseases. Prevention of age-related diseases means preventing the onset of various symptoms of age-related diseases in advance, or preventing the recurrence of symptoms after they have been suppressed. 【0039】Furthermore, compositions for removing senescent cells are expected to have applications in fields such as organ transplantation. For example, in transplant medicine, removing senescent cells from aged organs is expected to improve the success rate of transplantation. Similarly, by removing a patient's organ, such as a kidney, removing senescent cells, and re-transplanting it, it is expected that organ function will be restored or the disease condition will be improved. In autologous tissue transplantation in elderly patients, removing senescent cells is also expected to improve the prognosis. Moreover, its application to stem cell transplantation is also expected. Stem cells obtained from elderly patients have reduced proliferation efficiency due to the inclusion of senescent stem cells, which increases the risk of cancer after transplantation, so avoiding this is necessary. In cancer immunotherapy or certain infectious diseases, the aging of immune cells hinders treatment, so its application to aphoresis therapy, in which the patient's blood is taken out of the body, senescent cells are removed, and then returned to the patient, is also expected. 【0040】 The dosage of the senescent cell removal composition varies depending on the symptoms, age, administration method, dosage form, etc., but in typical cases, for adults, R １ -S-S-S-R ２ It is preferable to administer a compound represented by or a compound represented by formula (VI) in an amount of 0.5 to 50,000 mg per day, preferably 1 to 30,000 mg, in one or several divided doses per day for several consecutive days. 【0041】 The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples. 【0042】 In the following examples, the following compounds were used as test samples: pantetheine trisulfide (compound Ia) prepared by the method disclosed in WO2024 / 143400, glutathione trisulfide (compound IVa) prepared by the method described in WO2021 / 200487, and α-lipoic acid trisulfide (compound VIa) prepared by the method described in WO2022 / 045212. 【0043】 Manufacturing Example 1: Synthesis of pantetheine glutathione trisulfide (Ptt-SSS-G; compound (IIa)) (1) 【0044】475 mg (1.46 mmol, 1 eq.) of N,N'-Thiodhiphosphate and 4 mL (9 v / w) of isopropanol were charged into a 25 mL round-bottom flask. After confirming that the contents of the flask had dissolved, 450 mg (1.44 mmol, reference) of glutathione was charged, and after stirring for 1 hour, 400 mg (1.44 mmol, 1 eq.) of pantetheine was charged. After reacting at room temperature for 5 hours, the reaction mixture was concentrated under reduced pressure, and the concentrate was purified by column chromatography (ODS column, mobile phase: water / ethanol). Finally, the mixture was concentrated under reduced pressure at ambient temperature of 30°C and dried with an oil pump to obtain 47.5 mg (0.077 mmol, yield 5%, HPLC purity: 93%) of Ptt-SSS-G as a white solid. １ H-NMR: (D ２ O, 400MHz) δ (ppm) = 3.95-3.94 (m, 3H), 3.77 (dd, 1H, J = 6.4, 6.4Hz), 3.56-3.34 (m, 7H), 3.16 (dd, 2H, J = 6.4, 6.4Hz) ), 3.03 (dd, 2H, J = 6.4, 6.4Hz), 2.54-2.47 (m, 4H), 2.13 (ddd, 2H, J = 13.6, 7.2, 3.2Hz), 0.89 (s, 3H), 0.85 (s, 3H). ESI-TOF-MS: m / z 616.1788 ([M+H] ＋ ), calcd for[C ２１ H ３８ N ５ O １０ S ３ ] - 616.1781. 【0045】 Manufacturing Example 2: Synthesis of pantetheine glutathione trisulfide (Ptt-SSS-G; compound (IIa)) (2) 【0046】In a 300 mL square flask, 4.02 g (5.80 mmol, reference) of 80% pantethine aqueous solution, 4.18 g (5.80 mmol, 1 eq.) of oxidized glutathione hexahydrate, and 96 mL (24 v / w) of tap water were charged and cooled to 0°C. 4.16 g (12.8 mmol, 2.2 eq.) of Oxone® was added and the mixture was allowed to react for approximately 1 hour. Subsequently, an aqueous solution of sodium sulfide, prepared by dissolving 2.78 g (11.6 mmol, 2.0 eq.) of sodium sulfide notahydrate in 154 mL (19.2 v / w) of tap water, was added dropwise to the mixture. After refrigeration overnight, the reaction solution was purified by column chromatography (ODS column, mobile phase: water / ethanol). Finally, the material was concentrated under reduced pressure at an ambient temperature of 30°C and dried with an oil pump to obtain 1.12 g (1.82 mmol, yield 31%, HPLC purity: 97%) of Ptt-SSS-G as a white solid. 【0047】 Manufacturing Example 3: Synthesis of Pantetheine Cysteine ​​Trisulfide (Ptt-SSS-C; Compound (IIIa)) 【0048】 In a 20 mL test tube, 409 mg (1.70 mmol, reference) of cystine and 6 mL (14.7 v / w) of 1 mol / L hydrochloric acid were charged and cooled to 0°C. 610 mg (1.87 mmol, 1.1 eq.) of Oxone® was added and the mixture was allowed to react for approximately 1 hour. In another 20 mL test tube, 1.18 g (1.70 mmol, 1.0 eq.) of 80% pantethine aqueous solution and 6 mL (14.7 v / w) of tap water were charged and cooled to 0°C. 610 mg (1.87 mmol, 1.1 eq.) of Oxone® was added and the mixture was allowed to react for approximately 1 hour. The solutions from each reaction were mixed and cooled to 0°C. Next, an aqueous sodium sulfide solution, prepared by dissolving 817 mg (3.40 mmol, 2.0 eq.) of sodium sulfide notahydrate in 9.6 mL (23.5 v / w) of tap water, was added dropwise to the mixture. After refrigeration overnight, the reaction solution was purified by column chromatography (ODS column, mobile phase: water / ethanol). Finally, the solution was concentrated under reduced pressure at an ambient temperature of 30°C and dried with an oil pump to obtain 309 mg (0.7 mmol, yield 42%, HPLC purity: 97%) of white solid Ptt-SSS-Cys. １ H-NMR: (D ２O, 400MHz) δ (ppm) = 4.13 (dd, 1H, J = 8.0, 2.7Hz), 3.95 (s, 1H), 3.59-3.45 (m, 6H), 3.36 (d, 1H, J = 11.2Hz), 3.30 (dd, 1H, J=14.8, 8.0Hz), 3.06 (dd, 2H, J=6.4, 6.4Hz), 2.49 (dd, 2H, J=6.4, 6.4Hz), 0.89 (s, 3H), 0.85 (s, 3H). ESI-TOF-MS: m / z 430.1143 ([M+H] ＋ ), calcd for[C １４ H ２８ N ３ O ６ S ３ ] - 430.1140. 【0049】 Example 1: Comparative study of IL-1α production based on differences in the number of cell divisions of NB1RGB cells 1.1. Sample preparation for IL-1α expression analysis Human normal dermal fibroblasts (NB1RGB) with PDL12 (PDL: Population doubling level: number of cell divisions) and human senescent dermal fibroblasts (NB1RGB) with PDL50 or higher were cultured in Dulbecco's modified Eagle medium (DMEM; manufactured by Nissui) containing 10% fetal bovine serum (FBS). The cultured cells were harvested by trypsin treatment. The harvested cells were cultured at a rate of 8 × 10⁶ cells per well. ３ Seeds were sown in 96-well plates to form individual seeds, and incubated for 24 hours at 37°C and 5% CO2. ２ The samples were cultured in an incubator. Then, they were incubated in FBS-free DMEM for 48 hours at 37°C and 5% CO2. ２ The cells were cultured in an incubator. After culturing, the culture supernatant was collected. The culture supernatant was centrifuged, and this supernatant was used as the sample. 【0050】 1.2. IL-1α Expression Analysis: IL-1α in the culture supernatant is analyzed using ELISA (AuthentiKine). ＴＭ Measurements were performed using a Human IL-1 alpha ELISA Kit (Proteintech). The measurement wavelength was 450 nm, and the reference wavelength was 630 nm. The results are shown in Figure 1. As shown in Figure 1, fibroblasts that had undergone more than 50 divisions showed increased IL-1α production capacity and exhibited the phenotype of senescent cells. 【0051】 Example 2: Comparative study of cell viability of NB1RGB cells (PDL ≥ 50) 2.1. Cell viability study Human senescent dermal fibroblasts (NB1RGB) with a PDL of 50 or higher were cultured in DMEM containing 10% FBS. The cultured cells were harvested after trypsin treatment. The harvested cells were seeded in a 96-well plate and cultured at 37°C and 5% CO2 for 24 hours until 90% confluence was reached. ２ The samples were cultured in an incubator. Subsequently, the test samples (compounds (Ia), (IIa), (IIIa), (IVa), and (VIa)) were incubated in DMEM containing 200 μM, 400 μM, 600 μM, 800 μM, or 1000 μM for 24 hours at 37°C and 5% CO2. ２ The cells were cultured in an incubator. After culturing, the viability of human senescent skin fibroblasts was evaluated using Cell Counting Kit-8 (manufactured by Dojin Chemical Laboratories). The measurement wavelength was 450 nm (the measured absorbance was denoted as As), and the reference wavelength was 620 nm. 【0052】 2.2. Calculation of Cell Viability Human senescent skin fibroblasts were cultured in the same manner as in 2.1, except that no test sample was added, and the absorbance at a measurement wavelength of 450 nm was measured. The obtained absorbance was denoted as Ab, and the cell viability was calculated according to the following (Equation 1). Cell viability (%) = 100 × As / Ab ... (Equation 1) The results are shown in Figure 2. All trisulfide compounds reduced the cell viability of senescent cells in a concentration-dependent manner. 【0053】 Example 3: Comparative study of cell viability of NB1RGB cells (PDL=12) 3.1. Cell viability study Human normal dermal fibroblasts (NB1RGB) of PDL12 were cultured in DMEM containing 10% FBS. The cultured cells were harvested by trypsin treatment. The harvested cells were seeded in a 96-well plate and cultured at 37°C and 5% CO2 for 24 hours until 90% confluence was reached. ２ The samples were cultured in an incubator. Subsequently, the test samples (compound (Ia) and pantethine) were incubated in DMEM containing 200 μM, 400 μM, 600 μM, 800 μM, or 1000 μM for 24 hours at 37°C and 5% CO2. ２The cells were cultured in an incubator. After culturing, the viability of normal human skin fibroblasts was evaluated using Cell Counting Kit-8 (manufactured by Dojin Chemical Laboratories). The measurement wavelength (As) was 450 nm, and the reference wavelength was 620 nm. 【0054】 3.2. Calculation of Cell Viability Human normal skin fibroblasts were cultured in the same manner as in 3.1, except that no test sample was added, and the absorbance at a measurement wavelength of 450 nm was measured. The obtained absorbance was denoted as Ab, and the cell viability was calculated according to (Equation 1) above. The results are shown in Figure 3. 【0055】 Example 4: Comparative study of cell viability of NB1RGB cells (PDL ≥ 50) 4.1. Cell viability study Human senescent dermal fibroblasts (NB1RGB) with a PDL of 50 or higher were cultured in DMEM containing 10% FBS. The cultured cells were harvested by trypsin treatment. The harvested cells were seeded in a 96-well plate and cultured at 37°C and 5% CO2 for 24 hours until 90% confluence was reached. ２ The samples were cultured in an incubator. Subsequently, the test samples (compound (Ia) and pantethine) were incubated in DMEM containing 200 μM, 400 μM, 600 μM, 800 μM, or 1000 μM for 24 hours at 37°C and 5% CO2. ２ The cells were cultured in an incubator. After culturing, the viability of human senescent skin fibroblasts was evaluated using Cell Counting Kit-8 (manufactured by Dojin Chemical Laboratories). The measurement wavelength (As) was 450 nm, and the reference wavelength was 620 nm. 【0056】 4.2. Calculation of Cell Viability Human senescent skin fibroblasts were cultured in the same manner as in 4.1, except that no test sample was added, and the absorbance at a measurement wavelength of 450 nm was measured. The obtained absorbance was designated as Ab, and the cell viability was calculated according to (Equation 1) above. The results are shown in Figure 4. As can be seen from the results shown in Figures 3 and 4, the effect of trisulfide compounds on reducing cell viability is selective to senescent cells, and it was confirmed that pantethine, a disulfide compound, does not have this effect. 【0057】Example 5: Glutaminase 1 (GLS1) Inhibition Test in NB1RGB Cells (PDL ≥ 50) 5.1. Sample Preparation for GLS1 Expression Analysis Human senescent dermal fibroblasts (NB1RGB) with a PDL of 50 or higher were cultured in DMEM containing 10% FBS. The cultured cells were harvested by trypsin treatment. 3 × 10⁶ cells were harvested per well. ５ Seeds were sown in a 6-well plate to achieve the desired concentration, and incubated for 70 hours at 37°C and 5% CO2. ２ The samples were cultured in an incubator. Subsequently, the test samples (compound (Ia)) were incubated in DMEM containing 100 μM, 500 μM, or 1000 μM for 24 hours at 37°C and 5% CO2. ２ The cells were cultured in an incubator. After culturing, the supernatant was removed, washed twice with phosphate buffer, and the protein was extracted using RIPA buffer (Nacalai Tesque). The extracted protein was quantified using the Bradford method. 【0058】 5.2. Reference Cell Processing Human normal dermal fibroblasts (NB1RGB) of PDL12 were cultured using DMEM containing 10% FBS (manufactured by Nissui). The cultured cells were harvested by trypsin treatment. The harvested cells were divided into 1 x 10⁶ cells per dish. ５ Seeds were sown in 35 mm dishes to achieve the desired concentration, and incubated for 70 hours at 37°C and 5% CO2. ２ The cells were cultured in an incubator. Subsequent procedures were the same as in 5.1., except that the test sample was not added, to culture human normal skin fibroblasts and extract proteins. The extracted proteins were quantified using the Bradford method. 【0059】5.3. GLS1 Expression Analysis The amount of GLS1 in the samples was measured using ELISA with antibodies against GLS1. Two types of anti-GLS1 antibodies (Cell Signaling and Proteintech) were used as described in the datasheets. For the second antibody, an HRP-labeled antibody (SeraCare Life Sciences) was used, and after color development with 3,3',5,5'-tetramethylbenzidine (TMB), the reaction was stopped with 1N sulfuric acid. After stopping the reaction, the absorbance at a measurement wavelength of 450 nm was measured using a microplate reader. The reference wavelength was set to 620 nm. The relative amount of GLS1 when each test sample was added was calculated, with the amount of GLS1 in the control set to 1. The results are shown in Figure 5. It was confirmed that trisulfide compounds suppress the upregulation of GLS1 expression associated with cellular senescence in a concentration-dependent manner.

Claims

1. A compound represented by R １ -S-S-S-R ２ [where R １ and R ２ are each independently a group obtained by removing a sulfhydryl group from optionally protected pantetheine, a group obtained by removing a sulfhydryl group from optionally protected glutathione, or a group obtained by removing a sulfhydryl group from optionally protected cysteine], or a pharmaceutically acceptable salt thereof or a compound represented by formula (VI) [In formula (VI), X represents -OR ６１ or -NR ６２ R ６３ ; R ６１ represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms; R ６２ and R ６３ each independently represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and the alkyl group may have one or more substituents selected from the group consisting of an amino group and a carboxyl group], a composition for removing senescent cells containing a pharmaceutically acceptable salt thereof or a cyclodextrin inclusion complex thereof.

2. R １ -S-S-S-R ２ The compound represented by is the compound represented by formula (I). [R １１ and R １２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R １１ and R １２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R １３ and R １４ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R １３ and R １４ [Together, these are a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms], a compound represented by formula (II). [In formula (II), R ２１ and R ２２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R ２１ and R ２２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R ２３ and R ２４ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ２５ Compounds represented by formula (III), where is a protecting group for a hydrogen atom or an amino group. [In formula (III), R ３１ and R ３２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R ３１ and R ３２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R ３３ R is a protecting group for a hydrogen atom or a carboxyl group. ３４ Compounds represented by formula (IV), where is a protecting group for a hydrogen atom or an amino group. [In formula (IV), R ４１ , R ４２ , R ４３ and R ４４ are each independently a hydrogen atom or a protecting group for a carboxy group, and R ４５ and R ４６ are each independently a hydrogen atom or a protecting group for an amino group] or a compound represented by formula (V) [In formula (V), R ５１ and R ５２ are each independently a hydrogen atom or a protecting group for a carboxy group, and R ５３ and R ５４ are each independently a hydrogen atom or a protecting group for an amino group]. The composition according to claim 1 3. R １ -S-S-S-R ２ A compound represented by or a compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb), or formula (VIa). The composition according to claim 1.

4. The composition according to any one of claims 1 to 3, wherein the senescent cells are cells that highly express glutaminase 1.

5. The composition according to any one of claims 1 to 3, wherein the senescent cells are fibroblasts.

6. R １ -S-S-S-R ２ Compound represented by [R １ and R ２ Each of these is independently a group obtained by removing a sulfhydryl group from potentially protected pantetheine, a group obtained by removing a sulfhydryl group from potentially protected glutathione, or a group obtained by removing a sulfhydryl group from potentially protected cysteine, or a pharmaceutically acceptable salt thereof or a compound represented by formula (VI). [In equation (VI), X is -OR ６１ or -NR ６２ R ６３ R ６１ R represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms. ６２ and R ６３ A pharmaceutical composition for the treatment or prevention of age-related diseases, comprising a pharmaceutically acceptable salt thereof or a cyclodextrin inclusion complex thereof, wherein each of the following independently represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and the alkyl group may have one or more substituents selected from the group consisting of an amino group and a carboxyl group.

7. R １ -S-S-S-R ２ The compound represented by is the compound represented by formula (I). [R １１ and R １２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R １１ and R １２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R １３ and R １４ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R １３ and R １４ [Together, these are a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms], a compound represented by formula (II). [In formula (II), R ２１ and R ２２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R ２１ and R ２２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R ２３ and R ２４ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ２５ Compounds represented by formula (III), where is a protecting group for a hydrogen atom or an amino group. [In formula (III), R ３１ and R ３２ Each of these is independently a hydrogen atom, a C1-C9 alkyl group, or a C1-C5 acyl group, or R ３１ and R ３２ Together, they form a methylene group or carbonyl group which may have an alkyl group having 1 to 4 carbon atoms, and R ３３ R is a protecting group for a hydrogen atom or a carboxyl group. ３４ Compounds represented by formula (IV), where is a protecting group for a hydrogen atom or an amino group. [In formula (IV), R ４１ , R ４２ , R ４３ and R ４４ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ４５ and R ４６ [Each is independently a protecting group for a hydrogen atom or an amino group] or a compound represented by formula (V) [In formula (V), R ５１ and R ５２ Each of these is independently a protecting group for a hydrogen atom or a carboxyl group, and R ５３ and R ５４ The pharmaceutical composition according to claim 6, wherein each of the following is independently a protecting group for a hydrogen atom or an amino group.

8. R １ -S-S-S-R ２ The compound represented by or the compound represented by formula (VI) is a compound represented by formula (Ia), formula (Ib), formula (IIa), formula (IIIa), formula (IVa), formula (Va), formula (Vb) or formula (VIa) The pharmaceutical composition according to claim 6, wherein the composition is as described above 9. The pharmaceutical composition according to any one of claims 6 to 8, wherein the age-related disease is arteriosclerosis, arteriosclerotic cerebrovascular disease, osteoporosis, cataract, glaucoma, presbyopia, age-related macular degeneration, cerebral infarction, dementia, Parkinson's disease, cerebral hemorrhage, pulmonary fibrosis, chronic obstructive pulmonary disease, emphysema, type 2 diabetes, chronic renal failure, chronic kidney disease, cardiac hypertrophy, heart failure, hypertension, cirrhosis, fatty liver, non-alcoholic steatohepatitis, sarcopenia, skeletal muscle atrophy, emaciation, dyslipidemia, osteoarthritis, age-related lower back pain, age-related joint pain, frailty, age-related alopecia, age-related hearing loss, seborrheic dermatitis, pruritus, Hutchinson-Gilford progeria syndrome, Werner syndrome, Cockayne syndrome, or Rothmond-Thomson syndrome.

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