WO2023246925A1 - 酯类化合物及其应用 - Google Patents

酯类化合物及其应用 Download PDF

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WO2023246925A1
WO2023246925A1 PCT/CN2023/101960 CN2023101960W WO2023246925A1 WO 2023246925 A1 WO2023246925 A1 WO 2023246925A1 CN 2023101960 W CN2023101960 W CN 2023101960W WO 2023246925 A1 WO2023246925 A1 WO 2023246925A1
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substituted
group
unsubstituted
alkyl
scars
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PCT/CN2023/101960
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English (en)
French (fr)
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季银
王雪萍
翟慧
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南京瑞初医药有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/04—Ortho-condensed systems

Definitions

  • the present invention relates to the field of medicine, and in particular to a compound capable of targeting and killing senescent cells and its application in preventing or treating aging-related diseases.
  • SASP Senescence-Associated Secretory Phenotype
  • Senescent cells have many characteristics. Permanent cell cycle arrest is a common feature of all senescent cells and mainly depends on p16 and p21. Another broad-spectrum feature is the increased ⁇ -gal activity in senescent cells, that is, enzyme activity can also be detected in a non-optimal enzyme activity environment of pH 6.0, which is called senescence-associated ⁇ -galactosidase (Senescence- associated- ⁇ -galactosidase, SA- ⁇ -gal). Senescent cells cultured in vitro often exhibit enlargement and flattening of the nucleus and cytoplasm. Markers of aging also include activation of the DNA damage response, increased endoplasmic reticulum stress and ROS levels, and enhanced SASP secretion.
  • senescent cells are significantly induced to resist apoptosis, typically such as increased expression of BCL2/BCL-XL anti-apoptotic proteins. Therefore, senescent cells are also called zombie cells, which are old but do not die. Inducing cellular senescence can initially produce beneficial physiological effects by inhibiting tumor formation and promoting wound healing. Later studies found that the long-term presence of senescent cells in tissues may promote the occurrence of aging and age-related diseases, and even the inflammatory factors secreted by senescent cells will further Promote the occurrence of tumors.
  • SCAP Senescent Cell anti-Apoptotic Pathways
  • anti-apoptotic protein BCL2/BCL-XL Compared with normal cells, the expression of anti-apoptotic protein BCL2/BCL-XL in senescent cells is significantly increased, which antagonizes the activity of pro-apoptotic proteins such as Bax, making senescent cells resistant to apoptosis.
  • ABT263 navitoclax
  • A-1331852 is a small molecule inhibitor specific for Bcl-xL protein. In addition to its excellent killing effect on Bcl-xL-dependent cancer cells, A-1331852 also shows high selectivity in killing senescent cells. Although Bcl-xl can kill senescent cells, at a certain concentration It still has a strong killing effect on normal cells at low temperatures, so the safety of normal cells needs to be further improved.
  • the present invention provides a class of compounds that can selectively kill senescent cells and tumor cells within a larger safety window, while reducing the side effects of killing normal cells, and have Greater selectivity and security.
  • the first aspect of the application provides a compound represented by Formula I, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer or isotope labeled compounds,
  • R 1 is selected from substituted or unsubstituted C 1 -C 10 alkylene
  • R 2 is selected from -OC(O)- or -OC(O)-O-;
  • R 3 is selected from substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C2-C20 alkenyl, substituted or unsubstituted C2-C20 alkyne base, substituted or unsubstituted C6-C20 aryl group, substituted or unsubstituted C1-C20 heteroaryl group, substituted or unsubstituted C2-C20 heteroalicyclic group; alternatively, R 1 does not exist or is selected from substituted or unsubstituted Substituted C 1 -C 10 alkylene, R 2 is absent, and R 3 is selected from substituted or unsubstituted C2 - C20 heteroalicyclic group;
  • Each R a is independently selected from hydrogen, halogen, substituted or unsubstituted C1-C10 alkoxy, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C6-C20 aryl, substituted or unsubstituted C1 -C20 heteroaryl, nitro and cyano;
  • R b is selected from hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C6-C20 aryl and substituted or unsubstituted C1-C20 heteroaryl;
  • L is selected from substituted or unsubstituted C 1 -C 10 alkylene
  • R c is selected from substituted or unsubstituted adamantyl.
  • the substituent involved in the substitution is selected from the group consisting of halogen atoms, hydroxyl, thiol, amino, nitro, cyano, carboxyl, acyl, C1-C10 alkoxy, C6-C20 aryl, C1-C20 Heteroaryl, C2-C20 heteroalicyclic group, C1-C10 alkyl, C3-C8 cycloalkyl, C2-C8 alkenyl or C2-C8 alkynyl, halogen and/or hydroxyl substituted C1-C10 alkyl , halogen and/or hydroxyl substituted C3-C8 cycloalkyl, halogen and/or hydroxyl substituted C2-C8 alkenyl or halogen and/or hydroxyl substituted C2-C8 alkynyl, halogen and/or hydroxyl substituted C3 -C8 cycloalkyl.
  • the ring of the C2-C20 heteroalicyclic group optionally additionally contains 1 or 2 heteroatoms selected from N or O.
  • the C2-C20 heteroalicyclic group is optionally replaced by a halogen atom, a cyano group, a nitro group, a C6-C10 aryl group, a C1-C10 heteroaryl group, a C1-C6 alkoxy group, a C6 -C10 aryloxy group, C2-C10 heteroalicyclic group, amino group, hydroxyl group, mercapto group, carbonyl group, carboxyl group, acyl group, -NR 4 R 5 is substituted by one or more substituents, R 4 and R 5 are independently selected from hydrogen, C6-C10 aryl, C1-C10 heteroaryl, C1-C8 alkyl, C3-C8 Cycloalkyl, C2-C8 alkenyl and C2-C8 alkynyl, the aryl and heteroaryl groups are optionally substituted by halogen atoms, hydroxyl, mercapto, amino, nitro, cyano,
  • the compound is represented by Formula III, IV or V:
  • R 1 is selected from C 1 -C 10 alkylene independently selected from substituted or unsubstituted.
  • R 1 is selected from C 1 -C 6 alkylene independently selected from substituted or unsubstituted.
  • R 1 is selected from independently selected from substituted or unsubstituted C 1 -C 4 alkylene;
  • R a , R b , R 3 , R c , L are defined as in Formula I;
  • R 1 is absent or selected from C 1 -C 10 alkylene independently selected from substituted or unsubstituted.
  • R 1 is selected from C 1 -C 6 alkylene independently selected from substituted or unsubstituted. group, preferably, R 1 is selected from C 1 -C 4 alkylene independently selected from substituted or unsubstituted, R 3 is selected from substituted or unsubstituted C 2 -C 20 heteroalicyclic group, R a , R b , R
  • the definition of c and L is the same as formula I.
  • the C2-C20 heteroalicyclic group is optionally selected from the group consisting of halogen atoms, hydroxyl, thiol, amino, nitro, cyano, C1-C10 alkoxy, C1-C10 alkyl, C3 -C8 cycloalkyl, C2-C8 alkenyl, C2-C8 alkynyl, Substituted with a substituent, each R 5 is independently selected from hydrogen and C1-C6 alkyl.
  • the C2-C20 heteroalicyclic group is a C4-C8 heteroalicyclic group; the heteroalicyclic group is optionally selected from halogen, -NH2 , -OH, -NO2 , carbonyl , -CH 2 OH, carboxyl, methyl, ethyl, propyl, isopropyl, methoxy, ethoxy, propoxy or isopropoxy substituted.
  • the C2-C20 heteroalicyclic group is selected from the group shown below:
  • R' each independently represents no substituent, a single substituent or multiple substituents, and each substituent is independently selected from deuterium, hydroxyl, halogen, NH 2 , carboxyl (-COOH), C1-C6 alkyl, halogen-substituted C1-C6 alkyl, hydroxyl-substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, morpholine-substituted C1-C6 alkyl, -COO -C1-C6 alkyl, cyano, C1-C6 alkoxy, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, hydroxyl-substituted C3-C6 cycloalkyl, phenyl, benzyl base;
  • L 2 is a C1-C6 alkylene group that does not exist or is substituted by C1-C6 alkylene group, halogen, hydroxyl group, or C1-C6 alkoxy group, preferably methylene group, ethylene group, or propylene group;
  • R 6 is H, deuterium, halogen, hydroxyl, NH 2 , carboxyl (-COOH), -CONH 2 , sulfonic acid group (-SO 3 H), -SO 2 -C1-C6 alkyl group, C1-C6 alkyl, halogen-substituted C1-C6 alkyl, morpholine-substituted C1-C6 alkyl, -COO-C1-C6 alkyl, cyano, C1-C6 alkoxy, hydroxyl Substituted C1-C6 alkyl, amino-substituted C1-C6 alkyl, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, hydroxyl-substituted C3-C6 cycloalkyl, phenyl or benzyl base.
  • each R a is independently selected from hydrogen, halogen, substituted or unsubstituted C1-C10 alkoxy, substituted or unsubstituted C1-C10 alkyl. In some embodiments, each R a is independently selected from hydrogen, halogen, substituted or unsubstituted C1-C5 alkoxy, and substituted or unsubstituted C1-C5 alkyl.
  • each Ra is hydrogen
  • R b is selected from substituted or unsubstituted C1-C6 alkyl, preferably substituted or unsubstituted C1-C3 alkyl, more preferably methyl.
  • L is selected from substituted or unsubstituted C1-C6 alkylene, preferably substituted or unsubstituted C1-C3 alkylene. In some implementations, L is methylene.
  • the compound is represented by Formula II:
  • R 1 is selected from substituted or unsubstituted C 1 -C 10 alkylene
  • R 2 is selected from -OC(O)- or -OC(O)-O-;
  • R 3 is selected from substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C2-C20 alkenyl, substituted or unsubstituted C2-C20 alkyne base, substituted or unsubstituted C6-C20 aryl group, substituted or unsubstituted C1-C20 heteroaryl group, substituted or unsubstituted C2-C20 heteroalicyclic group.
  • R1 is substituted or unsubstituted C1-C6 alkylene, preferably substituted or unsubstituted C1-C3 alkylene, more preferably methylene.
  • R3 is selected from substituted or unsubstituted C1-C8 alkyl, substituted or unsubstituted C3-C10 cycloalkyl, substituted or unsubstituted C2-C8 alkenyl, substituted or unsubstituted C2-C8 alkynyl group, substituted or unsubstituted C6-C12 aryl group, substituted or unsubstituted C2-C12 heteroaryl group or substituted or unsubstituted C1-C12 heteroalicyclic group.
  • R3 is selected from substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C12 aryl, substituted or unsubstituted C1-C10 heteroaryl, substituted or unsubstituted C3-C8 ring Alkyl, or substituted or unsubstituted C1-C10 heteroalicyclic group.
  • substituted or unsubstituted means that one or more hydrogens in the described group can be substituted by substituents, and the substituents can be selected from halogen atoms, cyano groups, nitro groups, C6-C20 Aryl group, C1-C20 heteroaryl group, C1-C10 alkyl group, C1-C10 alkoxy group, C6-C20 aryloxy group, C1-C20 heteroalicyclic group, preferably C1-C10 heteroalicyclic group, amino group, Substituted with hydroxyl, thiol, phosphate group, -OC(O)R 6 , -ONR 6 R 7 , -NR 6 R 7 , R 6 and R 7 are independently selected from hydrogen, C6-C20 aryl, C1- C20 heteroaryl, C1-C8 alkyl, C3-C8 cycloalkyl base, C2-C8 alkenyl and C2-C8 alkyny
  • R3 is selected from the following groups:
  • the compound is selected from the following compounds:
  • the second aspect of the application provides a method for preventing or treating aging-related diseases, which method includes administering to a subject in need a therapeutically effective amount of the compound described in the first aspect of the application or a pharmaceutically acceptable amount thereof.
  • the diseases related to aging are diseases related to the accumulation of senescent cells.
  • the diseases are selected from the group consisting of idiopathic pulmonary fibrosis, pulmonary fibrosis, liver fibrosis, renal fibrosis, upper respiratory tract and pulmonary fibrosis caused by viruses.
  • Atrophy and fibrosis in various tissues Atrophy and fibrosis, brain and heart damage, treatment-related myelodysplastic syndromes, promyeloid syndrome, ataxia telangiectasia, Fanconi anemia, Friedreich's ataxia, congenital angle
  • dysplasia aplastic anemia, aneurysm, inflammatory bowel disease, lipoatrophy, renal transplant failure, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, glomerulosclerosis, and cancer or more.
  • the second aspect of the present application provides a compound or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, eutectic Use of isomers, stereoisomers or isotopically labeled compounds in the preparation of drugs for preventing or treating aging-related diseases, the compounds being the compounds described in the first aspect of the present application.
  • the diseases related to aging are diseases related to the accumulation of senescent cells.
  • the diseases are selected from the group consisting of idiopathic pulmonary fibrosis, pulmonary fibrosis, liver fibrosis, renal fibrosis, upper respiratory tract and pulmonary fibrosis caused by viruses.
  • Atrophy and fibrosis in various tissues Atrophy and fibrosis, brain and heart damage, treatment-related myelodysplastic syndromes, promyeloid syndrome, ataxia telangiectasia, Fanconi anemia, Friedreich's ataxia, congenital angle
  • dysplasia aplastic anemia, aneurysm, inflammatory bowel disease, lipoatrophy, renal transplant failure, sarcopenia, wound healing, alopecia, cardiomyocyte hypertrophy, glomerulosclerosis, and cancer or more.
  • the compound provided by the invention can reduce the side effects of killing normal cells, selectively kill senescent cells and tumor cells within a larger safety window, and has higher selectivity and safety.
  • Figure 1 shows the results of determination of carboxylesterase activity in normal and senescent cells.
  • Figure 2 shows the cell viability detection results of compounds on normal and senescent cells.
  • Figure 3 shows that intravitreal (IVT) administration of RZ001-RZ006 as well as RZ201 resulted in statistically significant improvements in avascular zone and vascular proliferation.
  • Figure 4 shows retinal and choroidal vascular leakage in a diabetic macular model after intravitreal (IVT) administration of RZ001, RZ002, and RZ201, which improved vascular permeability at this dose level.
  • Figure 5 shows statistically significant improvements in fluorescence intensity indicators in the CNV macular model following intravitreal (IVT) administration of RZ001, RZ002 and RZ201.
  • FIG. 6 shows that intraperitoneal (IP) administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in WBC, HYP and pathology scores in a pulmonary fibrosis (IPF) model.
  • IP intraperitoneal
  • Figure 7 shows that intra-articular administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in plantar mechanical pain threshold and safranin fast green stained pathology scores in an osteoarthritis (OA) model.
  • OA osteoarthritis
  • FIG 8 shows that intraperitoneal administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in A ⁇ 42 pathogenic protein indicators in Alzheimer's disease (AD) mouse models.
  • FIG. 9 shows that intraperitoneal (IP) administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels and pathology on a model of liver fibrosis.
  • IP intraperitoneal
  • Figure 10 shows that topical administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in skin epidermal thickness scores in a mouse model of natural aging.
  • FIG 11 shows that intraperitoneal (IP) administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in serum creatinine (Cr) and urea nitrogen (BUN) levels and pathology on a renal fibrosis model.
  • IP intraperitoneal
  • Figure 12 shows that administration of RZ001 resulted in a statistically significant improvement in the scar hyperplasia index of the rabbit ear hypertrophic scar model.
  • alkyl refers to an aliphatic hydrocarbon group, which may be branched or straight chain. Depending on the structure, an alkyl group can be a monovalent group or a bivalent group (i.e., an alkylene group). In the present invention, the alkyl group is preferably an alkyl group having 1 to 8 carbon atoms, more preferably a “lower alkyl group” having 1 to 6 carbon atoms, and even more preferably an alkyl group having 1 to 4 carbon atoms. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, etc.
  • alkyl includes all possible configurations and conformations of the alkyl group.
  • the "propyl” mentioned herein includes n-propyl and isopropyl
  • the "butyl” includes n-butyl. base, isobutyl and tert-butyl
  • "pentyl” includes n-pentyl, isopropyl, neopentyl, tert-pentyl, and pentyl-3-yl, etc.
  • alkoxy refers to -O-alkyl, where alkyl is as defined herein. Typical alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, etc.
  • cycloalkyl refers to a monocyclic or polycyclic group containing only carbon and hydrogen. Cycloalkyl groups include groups having 3 to 12 ring atoms. Depending on the structure, a cycloalkyl group can be a monovalent group or a bivalent group (eg, cycloalkylene). In the present invention, the cycloalkyl group is preferably a cycloalkyl group having 3 to 8 carbon atoms, and more preferably a “lower cycloalkyl group” having 3 to 6 carbon atoms.
  • cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and adamantane base.
  • aryl refers to an aryl ring in which each of the atoms making up the ring is a carbon atom.
  • Aryl rings can be composed of five, six, seven, eight, nine, or more than nine atoms.
  • Aryl groups may be optionally substituted. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, phenanthrenyl, anthracenyl, fluorenyl, and indenyl.
  • an aryl group can be a monovalent group or a bivalent group (i.e., arylene group).
  • heteroaryl refers to an aryl group that includes one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur.
  • the N-containing “heteroaryl” part refers to an aromatic group in which at least one skeleton atom in the ring is a nitrogen atom.
  • a heteroaryl group can be a monovalent group or a bivalent group (i.e., a heteroarylene group).
  • heteroaryl groups include, but are not limited to, pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazole base, isothiazolyl, pyrrolyl, quinolyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, isoindole Indolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazyl, benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl , naphthyridinyl and furopyr
  • alicyclic group or "cycloalkyl group” used herein refers to a non-aromatic ring formed by more than three carbon atoms. Two adjacent carbon atoms in the ring can be single bonds, double bonds or triple bonds. The number can be one or more.
  • Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclohexenyl, cyclopentenyl, cyclohexadienyl, and the like.
  • heterocycloalkyl or “heteroalicyclic” as used herein refers to a non-aromatic ring in which one or more ring-constituting atoms are heteroatoms selected from nitrogen, oxygen and sulfur. Heterocycloalkyl groups may be composed of three, four, five, six, seven, eight, nine or more than nine atoms. Heterocycloalkyl groups may be optionally substituted.
  • heteroalicyclic groups include, but are not limited to, lactams, lactones, cyclic imines, cyclic thioimines, cyclic carbamates, tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine, 1,3-dioxin, 1,3-dioxane, 1,4-dioxin, 1,4-dioxane, piperazine, 1,3-oxathiane, 1,4- Oxathiane, 1,4-oxathiane, tetrahydro-1,4-thiazine, 2H-1,2-oxazine, maleimide, succinimide, apeloline Bituric acid, thiobarbituric acid, dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane, hexahydro-1,3,5-triazine, tetrahydrothiophene, Tetrahydr
  • halogen refers to fluorine, chlorine, bromine and iodine.
  • salts formed by the compounds in the present invention also belong to the scope of the present invention. Unless otherwise stated, compounds in the present invention are understood to be Including its salts.
  • the term "salt” as used herein refers to an acidic or basic salt formed from an inorganic or organic acid and a base.
  • the compound of the present invention contains a basic moiety, which includes but is not limited to pyridine or imidazole, and when it contains an acidic moiety, including but is not limited to carboxylic acid, the zwitterion (“inner salt”) that may be formed is included in Within the scope of the term "salt”.
  • salts are preferred, although other salts are also useful, for example, in isolation or purification steps during preparation.
  • the compounds of the present invention may form salts, for example, compound I can be obtained by reacting with a certain amount of, for example, an equivalent amount of acid or base, salting out in a medium, or by freeze-drying in an aqueous solution.
  • the compounds of the present invention contain basic moieties, including but not limited to amines or pyridine or imidazole rings, which may form salts with organic or inorganic acids.
  • Typical acids that can form salts include acetates (eg, with acetic acid or trihaloacetic acids, such as trifluoroacetic acid), adipates, alginates, ascorbates, aspartates, and benzoates.
  • benzenesulfonate hydrogen sulfate, borate, butyrate, citrate, camphor salt, camphor sulfonate, cyclopentane propionate, diglycolate, dodecyl sulfate, Ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, enanthate, caproate, hydrochloride, hydrobromide, hydroiodide, glycolate (e.g., 2-hydroxyethanesulfonate), lactate, maleate, methanesulfonate, naphthalenesulfonate (e.g., 2-naphthalenesulfonate), nicotinate, nitrate, oxalic acid Salt, pectate, persulfate, phenylpropionate (such as 3-phenylpropionate), phosphate, picrate, pivalate, propionate, sal
  • Certain compounds of the invention may contain acidic moieties, including but not limited to carboxylic acids, which may form salts with various organic or inorganic bases.
  • Typical salts formed with bases include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, and salts formed with organic bases (such as organic amines), such as benzathine and dicyclohexylamine.
  • Hypamine salt with N,N-bis(dehydroabidyl)ethylenediamine
  • N-methyl-D-glucamine N-methyl-D-glucamide
  • tert-butyl Amines and salts formed with amino acids such as arginine, lysine, etc.
  • Basic nitrogen-containing groups can be combined with halide quaternary ammonium salts, such as small molecule alkyl halides (such as chlorides, bromides and iodides of methyl, ethyl, propyl and butyl), dialkyl sulfates (such as dimethyl sulfate, diethyl sulfate, dibutyl ester and dipentyl ester), long chain halides (such as decyl, dodecyl, tetradecyl and tetradecyl chlorides, bromides and iodide), aralkyl halides (such as benzyl and phenyl bromide), etc.
  • small halides such as chlorides, bromides and iodides of methyl, ethyl, propyl and butyl
  • dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl este
  • Prodrugs and solvates of the compounds of the present invention are also within the scope of the invention.
  • the term “prodrug” here refers to a compound that undergoes chemical transformation through metabolism or chemical processes to produce the compound, salt, or solvate of the present invention when treating related diseases.
  • “Solvate” refers to a solvent addition form containing a stoichiometric or non-stoichiometric amount of solvent. Some compounds tend to trap a fixed molar ratio of solvent molecules in the crystalline solid state, thus forming solvates. If the solvent is water, the solvate formed is a hydrate; if the solvent is alcohol, the solvate formed is an alcoholate. The combination of one or more water molecules with one molecule of a substance forms a hydrate, in which water retains its molecular state of H 2 O.
  • Non-limiting examples of solvates include ethanol solvates, acetone solvates, and the like.
  • the compounds, salts or solvates of the present invention may exist in tautomeric forms (eg amides and imine ethers). All such tautomers are part of the present invention.
  • All stereoisomers of the compounds are contemplated by the present invention.
  • the compounds of the invention may be independent stereoisomers that do not exist simultaneously with other isomers (e.g., have a specific activity as a pure or substantially pure optical isomer), or they may be mixtures, e.g. Racemates, or mixtures with all other stereoisomers or portions thereof.
  • the chiral center of the present invention has two configurations: S or R, which are defined as recommended by the International Union of Theoretical and Applied Chemistry (IUPAC) in 1974.
  • Racemic forms can be resolved by physical methods, such as fractional crystallization, or by fractional crystallization by derivatization to diastereoisomers, or by chiral column chromatography.
  • Individual optical isomers can be obtained from the racemate by suitable methods, including but not limited to traditional methods, such as salt formation with an optically active acid followed by recrystallization.
  • the weight content of the compounds in the present invention obtained by sequential preparation, separation and purification is equal to or greater than 90%, for example, equal to or greater than 95%, equal to or greater than 99% ("very pure" compounds), as described in the text List. Such "very pure” compounds of the invention are here also included as part of the invention.
  • Certain compounds of the present invention may exist in specific geometric or stereoisomeric forms.
  • the present invention encompasses all compounds, including their cis and trans isomers, R and S enantiomers, diastereomers, (D) isomers, (L) isomers, elimination Spin mixtures and other mixtures.
  • asymmetric carbon atoms can represent substituents, such as alkyl groups. All isomers, as well as mixtures thereof, are included in the present invention.
  • the mixture of isomers may contain the isomers in various ratios.
  • a mixture of only two isomers can have the following combinations: 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98: All ratios of isomers 2, 99:1, or 100:0 are within the scope of the invention. Similar ratios, as well as ratios for more complex mixtures of isomers that are readily understood by those of ordinary skill in the art, are also within the scope of the present invention.
  • the present invention also includes isotopically labeled compounds that are equivalent to the original compounds disclosed herein. In practice, however, it often occurs that one or more atoms are replaced by atoms with a different atomic weight or mass number.
  • isotopes of compounds that may be included in the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine isotopes such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O respectively. , 17 O, 31 P, 32 P, 35 S, 18 F and 36 Cl.
  • the compounds of the present invention or enantiomers, diastereomers, isomers, or pharmaceutically acceptable salts or solvates, which contain isotopes or other isotope atoms of the above compounds are within the scope of the present invention.
  • Certain isotopically labeled compounds of the present invention such as radioactive isotopes of 3 H and 14 C, are also included and are useful in tissue distribution experiments of drugs and substrates. Tritium, or 3H , and carbon-14, or 14C , are relatively easy to prepare and detect. It is the first choice among isotopes.
  • heavier isotope substitutions such as deuterium, i.e.
  • Isotopically labeled compounds can be prepared by general methods by replacing readily available isotopically labeled reagents with non-isotopic reagents, using the protocols disclosed in the Examples.
  • a synthesis of a specific enantiomer of the compound of the present invention it can be prepared by asymmetric synthesis, or derivatized with a chiral auxiliary, and the resulting diastereomeric mixture is separated and then the chiral auxiliary is removed. Pure enantiomer.
  • a suitable optically active acid or base can be used to form a diastereomeric salt with it, and then through separation, crystallization or chromatography, etc. After separation by conventional means, the pure enantiomers are obtained.
  • the compounds of the present invention may be provided with any number of substituents or functional groups to broaden their encompassing scope.
  • substituents or functional groups in general, whether the term “substituted” appears before or after the term “optional”, the general formula of the substituent included in the formulation of the present invention means that the substituent of the specified structure is used in place of the hydrogen radical. When multiple positions in a specific structure are substituted by multiple specific substituents, the substituents may be the same or different at each position.
  • substitution as used herein includes all permissible substitutions of organic compounds. Broadly speaking, permissible substituents include acyclic, cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic compounds.
  • heteroatoms such as nitrogen may have hydrogen substituents or any of the permissible organic compounds described above to supplement their valence. Furthermore, this invention is not intended to be limited in any way to the permitted substituted organic compounds.
  • the present invention considers that combinations of substituents and variable groups are excellent in the treatment of diseases in the form of stable compounds.
  • stable refers to a compound that is stable, detectable over a long enough period of time to maintain the structural integrity of the compound, and preferably effective over a long enough period of time, and is used herein for the above purposes.
  • SM8 chloromethyl butyrate
  • the external temperature of the oil bath was 50°C and continued to be heated and stirred for 3 hours. Dilute with ethyl acetate, wash with water and dry. Filter and concentrate. Column chromatography purification.
  • Step 1 Add potassium carbonate (16 mg) to a solution of A-1331852 (50 mg, 0.076 mmol, 1.00 eq) in DMF (1.0 ml), stir at room temperature for 30 min, and then add N- ⁇ [(2-methylpropane-2- (yl)oxy]carbonyl ⁇ -L-valine chloromethyl ester (24 mg, 1.2 eq). Oil bath external temperature Heat and stir at 50°C for 4 hours, then cool and add water, extract with ethyl acetate, wash with saturated brine, dry, concentrate, and obtain compound A (50 mg, yield: 74.1%) by column chromatography.
  • Step 2 Add 4M hydrochloric acid/dioxane solution (0.1 ml) to the solution of compound A (50 mg, 0.056 mmol) in dioxane (2 ml), and stir at room temperature for 1 hour. After diluting with dichloromethane, adjust the pH value to 7 with saturated sodium bicarbonate aqueous solution, extract, dry, concentrate, and purify by column chromatography.
  • Example 2 The esterase content in senescent cells is higher than that in normal cells.
  • This example selected human non-small cell lung cancer cell lines (A549 cells), human retinal pigment epithelial cells (RPE cells), and human pulmonary fibrosis cells (IMR90 cells). Detect whether the intracellular esterase content of cells after aging is greater than that of normal cells. To evaluate the advantages of prodrug release in senescent cells.
  • V sample added sample volume, 0.01mL; T: reaction time, 5 minutes; 200: total number of cells, 2 million; V sample total: volume of extraction solution added to cells, 0.2mL; V total: total volume of reaction system, 0.2mL; F: dilution factor.
  • the esterase content in senescent cells is significantly higher than that in normal cells.
  • Example 3 Prodrug compound has more specific killing effect on senescent cells than A-1331852
  • This example selected human non-small cell lung cancer cell line (A549 cells), human embryonic lung fibroblast cell line (IMR90 cells), human retinal pigment epithelial cells (RPE cells), mouse chondrocytes, and mouse skin fibroblasts. .
  • Senescent cells were obtained by etoposide induction (10 ⁇ M). The effects of the target compounds RZ001-RZ006 and RZ201 on the viability of the normal and aging cells were detected, thereby evaluating the specific killing effect of the compound prodrugs RZ001-RZ006 and RZ201 on the aging cells.
  • A-1331852 in addition to killing senescent cells, A-1331852 has a very obvious killing effect on normal cells.
  • Compounds RZ001-RZ006 and RZ201 in addition to killing senescent cells, have no obvious damage to normal cells. It shows that RZ001-RZ006 and RZ201 have better specificity and safety.
  • senescent cell clearance index (Senolytic index) on normal and senescent A549 cells was also calculated, see Table 4 below. The higher the index, the stronger the specificity of senescent cell removal and the better the safety.
  • Example 4 Efficacy in an animal model of hyperoxia-induced retinopathy
  • OIR oxygen-induced retinopathy
  • mice C57Bl/6 mouse pups and their CD1 adoptive mothers were exposed to a hyperoxic environment (75% O 2 ) from postnatal day 7 (P7) to postnatal day 12 (P12).
  • animals were injected intravitreally with 1 ⁇ l of the test composition (200uM compound) formulated in 1% DMSO, 10% Tween 80, 20% PEG-400 and returned to room air until day 17 (P17). Eyeballs were enucleated at P17 and retinas were dissected for vascular staining. To determine avascular or neovascular area, retinas were flattened and stained with isolectin B4 (IB4).
  • IB4 isolectin B4
  • Figure 3 shows that intravitreal (IVT) administration of RZ001-RZ006 as well as RZ201 resulted in statistically significant improvements in avascular zone and vascular proliferation.
  • Example 5 Efficacy in animal models of diabetes-induced retinopathy
  • the efficacy of the compounds of the present disclosure was studied in a mouse model of diabetic retinopathy by multiple administrations of streptozotocin (STZ).
  • C57BL/6J mice aged 6 to 7 weeks were weighed and their baseline blood glucose measured (Sino). Mice were injected intraperitoneally with STZ (Sigma-Alderich) at 55 mg/Kg for 5 consecutive days. Age-matched controls were injected with buffer only. One week after the last STZ injection, blood glucose was measured again.
  • STZ-treated diabetic C57BL/6J mice were injected intravitreally with 1 ⁇ l of test compound (200 uM in 0.015% polysorbate 80, 0.2% sodium phosphate, 0.75% sodium chloride, pH 7.2) 8 and 9 weeks after STZ administration. prepared as a suspension). Retinal Evans blue permeability assay was performed 10 weeks after STZ treatment.
  • Figure 4 shows retinal and choroidal vascular leakage after intravitreal (IVT) administration of RZ001, RZ002, and RZ201, all of which improved vascular permeability at this dose level.
  • Example 6 Efficacy in CNV in an animal model of wet-age-related macular degeneration
  • This example uses a CNV animal model to test the effect of the compounds of the present disclosure on wet-age-related macular degeneration.
  • mice Male mice were administered 0.5% probacaine hydrochloride (Alcon) for local anesthesia, and 1% tropicamide to dilate the mice's pupils.
  • Laser cauterization was performed using a laser photocoagulator (Novus Varia, LUMENIS) with a slit lamp delivery system. And artificially torn adhered glass coverslips are used as contact lenses. Bleeding spots caused by laser are not included in the statistics.
  • Figure 5 shows statistically significant improvements in fluorescence intensity indicators following intravitreal (IVT) administration of RZ001, RZ002 and RZ201.
  • Example 7 Efficacy in animal models of idiopathic pulmonary fibrosis
  • This example uses an idiopathic pulmonary fibrosis animal model to test the effect of the disclosed compounds on idiopathic pulmonary fibrosis.
  • anesthetized 6-week-old SD rats were fixed on the foam board with medical tape.
  • BLM 5mg/kg bleomycin
  • After administering the BLM solution immediately rotate and shake the foam board to distribute the BLM solution evenly in the lung tissue.
  • use surgical sutures to sew the skin on the surface of the trachea, and drip about 0.5-1.0ml of penicillin solution on the epidermis to prevent infection of the surgical wound.
  • IP intraperitoneal injection
  • the rats were euthanized by carbon dioxide, and the lung tissue was collected.
  • the left alveolar lavage fluid was collected for white blood cell (WBC) counting.
  • WBC white blood cell
  • HEP hydroxyproline
  • the remaining lung tissue was stained for pathological H&E. and Masson staining.
  • FIG. 6 shows that intraperitoneal (IP) administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in WBC, HYP and pathology scores on the IPF model.
  • IP intraperitoneal
  • This example uses an osteoarthritis animal model to test the effect of the disclosed compounds on osteoarthritis.
  • the osteoarthritis animal model was constructed through the following steps.
  • the mice were anesthetized with isoflurane and fixed.
  • the hair on the right hind limb of the mouse was shaved, the skin was wiped and disinfected with alcohol, and the skin at the joints was cut.
  • Under an optical microscope use a blade to cut the inside of the flexed right knee joint longitudinally. Do not cut the patellar ligament. Pull the patellar ligament to one side, and continue to use sterile cotton swabs to compress and stop bleeding, so that the joint cavity is fully exposed, and remove the patellar ligament in time. Add drops of normal saline to prevent the joint cavity from drying out. Use blunt forceps to gently separate excess muscle tissue to expose the meniscus ligament.
  • Figure 7 shows that intra-articular administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in plantar mechanical pain threshold and safranin fast green stained pathology scores in an osteoarthritis (OA) model.
  • OA osteoarthritis
  • Example 9 Efficacy in animal models of Alzheimer's disease
  • This example uses an Alzheimer's disease animal model to test the effect of the disclosed compounds on Alzheimer's disease.
  • the APP ⁇ PS1 transgenic mouse model (purchased from Saiye Biotechnology) was selected, and the compound of the present disclosure was administered by intraperitoneal injection for 11 consecutive weeks, twice a week. Observe the improvement of amyloid in the cerebral cortex of mice.
  • FIG 8 shows that intraperitoneal administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in A ⁇ 42 pathogenic protein indicators in AD mouse models.
  • Example 10 Efficacy in animal models of liver fibrosis
  • This example uses an animal model of liver fibrosis to test the effect of the compounds of the present disclosure on liver fibrosis. Briefly speaking, C57BL/6N mice, male, 7-8 weeks old, weighing 24-28g, were used. The mouse liver fibrosis model was induced by intraperitoneal injection of CCl4 solution at a dosage of 1 ml/kg, the frequency of administration was 2 times/week, and the modeling period was 6 weeks. Intraperitoneal injection (5 mg/kg) began in the third week of modeling and was administered twice a week. At the end of the experiment, serum was collected, liver tissue was taken, and Masson staining indicators were detected.
  • FIG. 9 shows that intraperitoneal (IP) administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels and pathology on a model of liver fibrosis.
  • IP intraperitoneal
  • Example 11 Effect on aging skin
  • This example used elderly female C57bl/6J mice (22 months old). The drug is administered by application through the skin. Administer twice a week for 6 weeks. Observe the improvement of skin epidermal thickness in aged mice.
  • Figure 10 shows that topical administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in skin epidermal thickness scores in a mouse model of natural aging.
  • Example 12 Efficacy in animal models of renal fibrosis
  • This example uses an animal model of renal fibrosis to test the effect of the compounds of the present disclosure on renal fibrosis. Briefly speaking, C57BL/6J mice, male, 7-8 weeks old, were used. After the mice were anesthetized, they were placed supine on a foam board and disinfected with iodophor. A longitudinal incision was made on the skin in the middle of the lower abdomen to find the white line of the abdominal muscles, where the abdominal muscles, fascia and peritoneal layers were dissected. After exposing the surgical field, use gauze soaked in saline to push the stomach and mesentery of the mouse to the right side to expose the kidneys.
  • FIG 11 shows that intraperitoneal (IP) administration of RZ001, RZ002 and RZ201 resulted in statistically significant improvements in serum creatinine (Cr) and urea nitrogen (BUN) levels and pathology on a renal fibrosis model.
  • IP intraperitoneal
  • This example uses a rabbit ear hypertrophic scar animal model to test the effect of the disclosed compounds on hypertrophic scars. To put it simply, 10 New Zealand white rabbits, male, 3 months old, were selected. After New Zealand rabbits were anesthetized with sodium pentobarbital, six circular full-thickness wounds with a diameter of 10 mm were made on the ventral surface of each ear by removing the epidermis, dermis, and perichondrium to exposed cartilage. Rabbits were randomly divided into control group and RZ001 experimental group.
  • Figure 12 shows that administration of RZ001 resulted in a statistically significant improvement in the scar hyperplasia index of the rabbit ear hypertrophic scar model.

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Abstract

本发明公开了一种化合物或其药学上可接受的盐、溶剂化物、水合物、多晶型物、共晶、互变异构体、立体异构体或同位素标记化合物。该化合物如式（I）所示。本发明还公开了该化合物在制备用于防或治疗与衰老相关疾病的药物中的应用。

Description

酯类化合物及其应用 技术领域
本发明涉及医药领域，尤其涉及一种能够靶向杀伤衰老细胞的化合物及其在预防或治疗与衰老相关疾病中应用。
背景技术
随着年龄的增长，机体生物学功能逐渐衰退，个体老化的过程往往伴随着各种老年性疾病的发生。因此，开发有效的衰老干预策略对于延长老年群体的健康寿命、降低衰老相关疾病的发病率意义重大。衰老细胞在体内的积累是导致个体衰老和衰老相关疾病发生的重要驱动力：一方面，一些增殖细胞或者干细胞的衰老会导致个体再生能力的下降，影响机体的恢复能力和正常功能；更为重要的是，衰老细胞分泌大量炎症因子，称为衰老相关的分泌表型(Senescence-Associated Secretory Phenotype，SASP)，造成慢性炎性微环境，加速机体衰老进程，并促进与年龄有关疾病的发生。研究表明，通过遗传学方法或者化学小分子特异清除衰老细胞可以降低慢性炎症，提高组织修复能力，改善如骨关节炎和特发性肺纤维化等老年疾病症状，并缓解衰老生物体生理功能的下降。因此，选择性清除衰老细胞是治疗衰老相关疾病和改善老年个体生理机能的有前景的途径。
衰老细胞具有诸多特征，永久性细胞周期停滞是所有衰老细胞共有的特征，主要依赖于p16和p21。另一个广谱的特征是衰老细胞中的β-gal活性增加，即在pH 6.0的非最佳酶活性环境也可以检测到酶活性，被称为衰老相关的β-半乳糖苷酶(Senescence-associated-β-galactosidase,SA-β-gal)。体外培养的衰老细胞通常会表现出细胞核和细胞质扩大，形状变平。衰老的标志物还包括DNA损伤反应的激活，内质网应激和ROS水平增加，SASP分泌增强。此外，衰老细胞中的抗凋亡组分被显著诱导用以抵抗细胞凋亡，典型的如BCL2/BCL-XL抗凋亡蛋白表达增高。因此，衰老细胞又被称为僵尸细胞，老而不死。诱导细胞衰老最初可以通过抑制肿瘤形成和促进伤口愈合产生有益的生理作用，后来研究发现衰老细胞在组织中的长期存在可能促进衰老与年龄有关疾病的发生，甚至衰老细胞分泌的炎症因子也会进一步促进肿瘤的发生。
衰老细胞的积累与衰老及衰老相关疾病发生的因果关系通过遗传学途径已经得到明确解析。梅奥诊所Deursen教授研究组在2011年构建p16启动子驱动的INK-ATTAC转基因小鼠，该系统仅在p16阳性衰老细胞中选择性表达FKBP-CASP8融合蛋白。经小分子AP20187处理后，FKBP-CASP8融合蛋白二聚化并诱导p16阳性的衰老细胞发生凋亡。在Bub1b突变型早衰小鼠中利用此方法间歇性清除衰老细胞，可以观察到肌肉减少症、白内障和脂肪营养不良得到显著改观。在2016年的研究中，研究人员通过每两周对自然衰老的INK-ATTAC小鼠进行AP20187给药以清除衰老细胞，结果显示p16阳性衰老细胞的清除减弱了部分组织器官(包括肾脏、心脏和脂肪)年龄相关的退化，并延长了中位寿命。Kirland教授课题组将仅仅少量的衰老细胞植入受体小鼠体内，便可以驱动年轻小鼠持续的身体机能障碍，从而导致功能下降和死亡率增加。这些研究成果从不同角度证明在体内清除衰老细胞可以延长寿命以及延缓衰老相关疾病的发生，为靶向杀伤衰老细胞实现抗衰老以及治疗衰老相关的疾病的策略提供了理论依据。
目前已报道多个可选择性清除衰老细胞的小分子化合物，该类小分子也被称为“senolytics”。由于衰老细胞的异质性，这些senolytics虽然在一定程度上能实现对衰老细胞的选择性杀伤，但是在降低毒性以及提高杀伤的特异性方面仍有提升空间。早期的senolytics大多靶向衰老细胞抗凋亡途径(Senescent Cell anti-Apoptotic Pathways，SCAP)中的重要组分，即通过阻断衰老细胞的抗凋亡信号进而诱导其死亡。与正常细胞相比，衰老细胞中抗凋亡蛋白BCL2/BCL-XL表达显著升高，拮抗了Bax等促凋亡蛋白活性，致使衰老细胞可抵抗凋亡。作为经典抗凋亡蛋白BCL2/BCL-XL的双重抑制剂，ABT263(navitoclax)随后被证明可通过特异诱导衰老细胞凋亡。
A-1331852是Bcl-xL蛋白特异性的小分子抑制剂。A-1331852除对依赖Bcl-xL的癌细胞具有非常好的杀伤效果以外，其对衰老细胞的杀伤也表现出较高的选择性。尽管Bcl-xl对衰老细胞有杀伤，但在一定浓 度下对正常细胞仍旧有很强杀伤作用，因此对于正常细胞的安全性有待进一步提高。
发掘衰老细胞与非衰老细胞之间存在的差异是实现对衰老细胞靶向识别和杀伤的基础。发明人通过研究发现衰老细胞中的羧酸酯酶活性显著高于非衰老细胞。因此，羧酸酯酶有可能成为衰老细胞的潜在标记物，为基于该类酶激活的前药策略运用提供了理论基础。目前缺乏基于A-1331852修饰改造的Bcl-xL抑制剂，可以更高效特异的诱导衰老细胞凋亡，并利用该方法在哺乳动物中抗衰老以及治疗衰老相关疾病。本发明利用衰老细胞中酯酶活性高的特征，设计了新型前药化合物，满足了这些需求并提供了相关的优势。
发明内容
为了解决现有技术中存在的至少一个技术问题，本发明提供了一类化合物，其能够在更大的安全窗口内选择性杀伤衰老细胞和肿瘤细胞，同时降低了对正常细胞的杀伤副作用，具备更高的选择性和安全性。
本申请第一方面提供了一种式I所示的化合物，或其药学上可接受的盐、溶剂化物、水合物、多晶型物、共晶、互变异构体、立体异构体或同位素标记化合物，
式I中，
R1选自取代或未取代的C1-C10亚烷基；
R2选自-O-C(O)-或者-O-C(O)-O-；
R3选自取代或未取代的C1-C10链烷基、取代或未取代的C3-C20环烷基、取代或未取代的C2-C20链烯基、取代或未取代的C2-C20链炔基、取代或未取代的C6-C20芳基、取代或未取代的C1-C20杂芳基、取代或未取代的C2-C20杂脂环基；或者，R1不存在或者选自取代或未取代的C1-C10亚烷基，R2不存在，并且R3选自取代或未取代的C2-C20杂脂环基；
各Ra独立选自氢、卤素、取代或未取代的C1-C10烷氧基、取代或未取代的C1-C10烷基、取代或未取代的C6-C20芳基、取代或未取代的C1-C20杂芳基、硝基和氰基；
Rb选自氢、取代或未取代的C1-C10烷基、取代或未取代的C6-C20芳基和取代或未取代的C1-C20杂芳基；
L选自取代或未取代的C1-C10亚烷基；
Rc选自取代或未取代的金刚烷基。
在一些实施方式中，所述取代涉及的取代基选自卤素原子、羟基、巯基、氨基、硝基、氰基、羧基、酰基、C1-C10烷氧基、C6-C20芳基、C1-C20杂芳基、C2-C20杂脂环基、C1-C10烷基、C3-C8环烷基、C2-C8链烯基或C2-C8炔基、卤素和/或羟基取代的C1-C10烷基、卤素和/或羟基取代的C3-C8环烷基、卤素和/或羟基取代的C2-C8链烯基或卤素和/或羟基取代的C2-C8炔基、卤素和/或羟基取代的C3-C8环烷基。
在一些实施方式中，C2-C20杂脂环基的环中任选地额外含有1或2个选自N或O的杂原子。
在一些实施方式中，所述C2-C20杂脂环基任选地被卤素原子、氰基、硝基、C6-C10芳基、C1-C10杂芳基、C1-C6链烷氧基、C6-C10芳氧基、C2-C10杂脂环基、氨基、羟基、巯基、羰基、羧基、酰基、 -NR4R5中的一个或两个以上的取代基取代，R4和R5独立选自氢、C6-C10芳基、C1-C10杂芳基、C1-C8链烷基、C3-C8环烷基、C2-C8链烯基和C2-C8炔基，所述芳基和杂芳基任选被卤素原子、羟基、巯基、氨基、硝基、氰基、羧基、酰基、C1-C8烷氧基、C6-C10芳基、C1-C10杂芳基、C2-C10杂脂环基、C1-C8烷基、C3-C8环烷基、C2-C6链烯基或C2-C8炔基取代，任选地，其中至少两个位置的取代基共同构成C3-C10脂肪环、C2-C10杂脂环、C6-C10芳环或C1-C10杂芳环。
在一些实施方式中，所述化合物如式III、IV或V所示：
式III和IV中，R1选自独立选自取代或未取代的C1-C10亚烷基，优选地，R1选自独立选自取代或未取代的C1-C6亚烷基，优选地，R1选自独立选自取代或未取代的C1-C4亚烷基；Ra、Rb、R3、Rc、L的定义同式I；
式V中，R1不存在或者选自独立选自取代或未取代的C1-C10亚烷基，优选地，R1选自独立选自取代或未取代的C1-C6亚烷基，优选地，R1选自独立选自取代或未取代的C1-C4亚烷基，R3选自取代或未取代的C2-C20杂脂环基，Ra、Rb、Rc、L的定义同式I。
在一些实施方式中，所述C2-C20杂脂环基任选地被选自卤素原子、羟基、巯基、氨基、硝基、氰基、C1-C10烷氧基、C1-C10烷基、C3-C8环烷基、C2-C8链烯基、C2-C8炔基、 的取代基取代，各R5独立选自氢和C1-C6烷基。
在一些实施方式中，所述C2-C20杂脂环基是C4-C8杂脂环基；所述杂脂环基任选地被选自卤素、-NH2、-OH、-NO2、羰基、-CH2OH、羧基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基或异丙氧基取代。
在一些实施方式中，所述C2-C20杂脂环基选自下组所示基团：
R’各自独立地表示无取代基、单个取代基或多个取代基，各取代基独立地选自氘、羟基、卤素、NH2、羧基(-COOH)、C1-C6链烷基、卤素取代的C1-C6链烷基、羟基取代的C1-C6链烷基、氨基取代的C1-C6链烷基、吗啉取代的C1-C6链烷基、-COO-C1-C6链烷基、氰基、C1-C6链烷氧基、C3-C6环烷基、卤素取代的C3-C6环烷基、羟基取代的C3-C6环烷基、苯基、苄基；
L2为不存在或C1-C6亚烷基、卤素、羟基、C1-C6烷氧基取代的C1-C6亚烷基，优选亚甲基、亚乙基、亚丙基；
R6为H、氘、卤素、羟基、NH2、羧基(-COOH)、-CONH2、磺酸基(-SO3H)、-SO2-C1-C6链烷基、C1-C6链烷基、卤素取代的C1-C6链烷基、吗啉取代的C1-C6链烷基、-COO-C1-C6链烷基、氰基、C1-C6链烷氧基、羟基取代的C1-C6链烷基、氨基取代的C1-C6链烷基、C3-C6环烷基、卤素取代的C3-C6环烷基、羟基取代的C3-C6环烷基、苯基或苄基。
在一些实施方式中，各Ra独立选自氢、卤素、取代或未取代的C1-C10烷氧基、取代或未取代的C1-C10烷基。在一些实施方式中，各Ra独立选自氢、卤素、取代或未取代的C1-C5烷氧基和取代或未取代的C1-C5烷基。
在一些实施方式中，各Ra均为氢。
在一些实施方式中，Rb选自取代或未取代的C1-C6烷基，优选取代或未取代的C1-C3烷基，更优选甲基。
在一些实施方式中，L选自取代或未取代的C1-C6亚烷基，优选取代或未取代的C1-C3亚烷基。在一些实施中，L是亚甲基。
在一些实施方式中，所述化合物如式II所示：
式II中，对R1、R2、R3的限定同式I，即：
R1选自取代或未取代的C1-C10亚烷基；
R2选自-O-C(O)-或者-O-C(O)-O-；
R3选自取代或未取代的C1-C10链烷基、取代或未取代的C3-C20环烷基、取代或未取代的C2-C20链烯基、取代或未取代的C2-C20链炔基、取代或未取代的C6-C20芳基、取代或未取代的C1-C20杂芳基、取代或未取代的C2-C20杂脂环基。
在一些实施方式中，R1为取代或未取代的C1-C6亚烷基，优选为取代或未取代的C1-C3亚烷基，更优选为亚甲基。
在一些实施方式中，R3选自取代或未取代的C1-C8链烷基、取代或未取代的C3-C10环烷基、取代或未取代的C2-C8链烯基、取代或未取代的C2-C8链炔基、取代或未取代的C6-C12芳基、取代或未取代的C2-C12杂芳基或者取代或未取代的C1-C12杂脂环基。
优选地，R3选自取代或未取代的C1-C6烷基，取代或未取代的C6-C12芳基，取代或未取代的C1-C10杂芳基，取代或未取代的C3-C8环烷基，或者取代或未取代的C1-C10杂脂环基。
在本申请中，“取代或未取代”是指其描述的基团中的一个或多个氢可以被取代基取代，所述取代基可以选自卤素原子、氰基、硝基、C6-C20芳基、C1-C20杂芳基、C1-C10链烷基、C1-C10链烷氧基、C6-C20芳氧基、C1-C20杂脂环基优选C1-C10杂脂环基、氨基、羟基、巯基、磷酸酯基、-OC(O)R6、-ONR6R7、-NR6R7的取代基取代，R6和R7独立选自氢、C6-C20芳基、C1-C20杂芳基、C1-C8链烷基、C3-C8环烷 基、C2-C8链烯基和C2-C8链炔基，所述芳基和杂芳基任选被卤素原子、羟基、巯基、氨基、硝基、氰基、羧基、酰基、C1-C6烷氧基、C6-C20芳基、C1-C20杂芳基、C2-C20杂脂环基、C1-C10烷基、C3-C8环烷基、C2-C8链烯基或C2-C8链炔基取代，其中任选至少两个位置的取代基共同构成脂肪环例如C3-C20脂肪环、杂脂环例如C2-C20脂肪环、芳环例如C6-C20芳环或杂芳环例如C1-C20杂芳环。
在一些实施方式中，R3选自以下基团：
在一些实施方式中，所述化合物选自以下化合物：




本申请第二方面提供了一种用于预防或治疗与衰老相关疾病的方法，该方法包括向有需要的受试者施用治疗有效量的本申请第一方面所述的化合物或其药学上可接受的盐、溶剂化物、水合物、多晶型物、共晶、互变异构体、立体异构体或同位素标记化合物。
具体地，与衰老相关疾病是衰老细胞的积累相关的疾病，优选地，所述疾病选自特发性肺纤维化、肺纤维化、肝纤维化、肾纤维化、病毒导致的上呼吸道和肺部的炎症和组织纤维化及萎缩、囊性纤维化、骨髓纤维化、心肌纤维化、皮肤纤维化、间质性肺疾病、纤维化的胰腺炎、早产儿视网膜病变、黄斑变性、糖尿病性黄斑水肿、糖尿病性视网膜病变、年龄相关性黄斑变性、湿性年龄相关黄斑变性、干性年龄相关黄斑变性、青光眼、镰状细胞视网膜病变、缺血性动脉炎性神经病变、干燥性角膜炎、Fuch氏角膜营养不良、老视、白内障、变性玻璃体病症，包括玻璃体黄斑牵引综合征、黄斑裂孔、视网膜撕裂、视网膜剥离、增殖性玻璃体视网膜病变、骨关节炎、椎间盘突出、骨质疏松、阿尔兹海默症、帕金森病、动脉粥样硬化、慢性阻塞性肺病、糖尿病、糖尿病肾病、瘢痕、表浅性瘢痕或扁平瘢痕、索条状瘢痕或挛缩性瘢痕、蹼状瘢痕、凹陷性瘢痕、萎缩性瘢痕、桥状瘢痕和赘状瘢痕、增生性瘢痕、瘢痕疙瘩、瘢痕癌、硬皮病、硬斑病、带状硬皮病、点滴状硬皮病、肢端硬皮病、弥漫性硬皮病、CREST综合征、急性冠状动脉综合征、心肌梗塞、中风、高血压、肥胖、脂肪功能障碍、冠状动脉疾病、脑血管疾病、牙周病、癌症治疗相关的残疾例如各种组织中的萎缩和纤维化、脑和心脏损及治疗相关的骨髓增生异常综合征、早幼粒综合征、共济失调毛细血管扩张症、范可尼贫血、弗里德赖希共济失调、先天性角化不良、再生障碍性贫血、动脉瘤、炎症性肠病、脂肪萎缩、肾脏移植手术失败、肌肉衰减征、伤口愈合、秃头症、心肌细胞肥大、肾小球硬化症、和癌症中的一种或多种。
本申请第二方面提供了一种化合物或其药学上可接受的盐、溶剂化物、水合物、多晶型物、共晶、互 变异构体、立体异构体或同位素标记化合物在制备用于防或治疗与衰老相关疾病的药物中的应用，所述化合物为如本申请第一方面所述的化合物。
具体地，与衰老相关疾病是衰老细胞的积累相关的疾病，优选地，所述疾病选自特发性肺纤维化、肺纤维化、肝纤维化、肾纤维化、病毒导致的上呼吸道和肺部的炎症和组织纤维化及萎缩、囊性纤维化、骨髓纤维化、心肌纤维化、皮肤纤维化、间质性肺疾病、纤维化的胰腺炎、早产儿视网膜病变、黄斑变性、糖尿病性黄斑水肿、糖尿病性视网膜病变、年龄相关性黄斑变性、湿性年龄相关黄斑变性、干性年龄相关黄斑变性、青光眼、镰状细胞视网膜病变、缺血性动脉炎性神经病变、干燥性角膜炎、Fuch氏角膜营养不良、老视、白内障、变性玻璃体病症，包括玻璃体黄斑牵引综合征、黄斑裂孔、视网膜撕裂、视网膜剥离、增殖性玻璃体视网膜病变、骨关节炎、椎间盘突出、骨质疏松、阿尔兹海默症、帕金森病、动脉粥样硬化、慢性阻塞性肺病、糖尿病、糖尿病肾病、瘢痕、表浅性瘢痕或扁平瘢痕、索条状瘢痕或挛缩性瘢痕、蹼状瘢痕、凹陷性瘢痕、萎缩性瘢痕、桥状瘢痕和赘状瘢痕、增生性瘢痕、瘢痕疙瘩、瘢痕癌、硬皮病、硬斑病、带状硬皮病、点滴状硬皮病、肢端硬皮病、弥漫性硬皮病、CREST综合征、急性冠状动脉综合征、心肌梗塞、中风、高血压、肥胖、脂肪功能障碍、冠状动脉疾病、脑血管疾病、牙周病、癌症治疗相关的残疾例如各种组织中的萎缩和纤维化、脑和心脏损及治疗相关的骨髓增生异常综合征、早幼粒综合征、共济失调毛细血管扩张症、范可尼贫血、弗里德赖希共济失调、先天性角化不良、再生障碍性贫血、动脉瘤、炎症性肠病、脂肪萎缩、肾脏移植手术失败、肌肉衰减征、伤口愈合、秃头症、心肌细胞肥大、肾小球硬化症、和癌症中的一种或多种。
本发明提供的化合物能够降低了对正常细胞的杀伤副作用，在更大的安全窗口内选择性杀伤衰老细胞和肿瘤细胞，具备更高的选择性和安全性。
附图说明
图1显示对正常及衰老的细胞中羧酸酯酶活性的测定结果。
图2显示化合物对正常和衰老细胞的细胞活力检测结果。
图3显示玻璃体内(IVT)施用RZ001-RZ006以及RZ201导致无血管区和血管增生的统计学显著改善。
图4显示玻璃体内(IVT)施用RZ001，RZ002及RZ201后的糖尿病黄斑模型视网膜和脉络膜血管渗漏在此剂量水平均可改善血管通透性。
图5显示玻璃体内(IVT)施用RZ001，RZ002及RZ201后的CNV黄斑模型荧光强度指标的统计学显著改善。
图6显示腹腔注射(IP)施用RZ001，RZ002及RZ201导致肺纤维化(IPF)模型上WBC，HYP和病理学评分的统计学显著改善。
图7显示关节腔注射施用RZ001，RZ002及RZ201导致骨关节炎(OA)模型上足底机械疼痛阈值和番红固绿染色的病理学评分的统计学显著改善。
图8显示腹腔注射施用RZ001，RZ002及RZ201导致阿尔兹海默症(AD)小鼠模型上Aβ42致病蛋白指标的统计学显著改善。
图9显示腹腔注射(IP)施用RZ001，RZ002和RZ201导致肝纤维化模型上血清中谷草转氨酶(AST)和谷丙转氨酶(ALT)水平和病理学的统计学显著改善。
图10显示涂抹施用RZ001，RZ002和RZ201导致自然衰老小鼠模型皮肤表皮厚度评分的统计学显著改善。
图11显示腹腔注射(IP)施用RZ001，RZ002和RZ201导致肾纤维化模型上血清中肌酐(Cr)和尿素氮(BUN)水平和病理学的统计学显著改善。
图12显示施用RZ001导致兔耳增生性瘢痕模型瘢痕增生指数统计学显著改善。
具体实施方式
以下结合实施例和附图对本发明进行详细说明：本实施例在以本发明为技术方案为前提下进行实施，给出了详细实施方案和过程，但本发明提供的实施方案是示例性的，旨在用于解释本发明，而不能理解为对本发明的限制。下述的实施例未注明的条件和方法等均按常规进行。
定义
术语“烷基”是指脂肪族烃基团，可以是支链或直链的烷基。根据结构，烷基可以是单价基团或双价基团(即亚烷基)。在本发明中，烷基优选是具有1-8个碳原子的烷基，更优选具有1-6个碳原子的“低级烷基”，甚至更优选具有1-4个碳原子的烷基。典型的烷基包括但不限于甲基、乙基、丙基、丁基、戊基、己基等。应理解，本文提到的“烷基”包括可能存在的所有构型和构象的该烷基，例如本文提到的“丙基”包括正丙基和异丙基，“丁基”包括正丁基、异丁基和叔丁基，“戊基”包括正戊基、异丙基、新戊基、叔戊基、和戊-3-基等。
术语“烷氧基”是指-O-烷基，其中烷基如本文中定义。典型的烷氧基包括但不限于甲氧基、乙氧基、丙氧基、丁氧基、戊氧基、己氧基等。
术语“环烷基”是指单环或多环基，其仅含有碳和氢。环烷基包括具有3-12个环原子的基团。根据结构，环烷基可以是单价基团或双价基团(例如亚环烷基)。在本发明中，环烷基优选是具有3-8个碳原子的环烷基，更优选具有3-6个碳原子的“低级环烷基”。环烷基的例子包括但不限于，环丙基、环丁基、环戊基、环己基、环庚基、环辛基、环戊烯基、环己烯基、环庚烯基和金刚烷基。
术语“芳基”是指芳香基环中每一个构成环的原子都是碳原子。芳基环可以由五、六、七、八、九或多于九个原子构成。芳基可以是任选取代的。芳基的实例包括但不限于苯基、萘基、菲基、蒽基、芴基和茚基。根据结构，芳基可以是单价基团或双价基团(即亚芳基)。
术语“杂芳基”是指芳基中包括一个或多个选自氮、氧和硫的环杂原子。含N“杂芳基”部分是指芳香基中环上至少有一个骨架原子是氮原子。根据结构，杂芳基可以是单价基团或双价基团(即亚杂芳基)。杂芳基的实例包括但不限于吡啶基、咪唑基、嘧啶基、吡唑基、三唑基、吡嗪基、四唑基、呋喃基、噻吩基、异噁唑基、噻唑基、噁唑基、异噻唑基、吡咯基、喹啉基、异喹啉基、吲哚基、苯并咪唑基、苯并呋喃基、吲唑基、吲嗪基、酞嗪基、哒嗪基、异吲哚基、蝶啶基、嘌呤基、噁二唑基、噻二唑基、呋咱基、苯并呋咱基、苯并噻吩基、苯并噻唑基、苯并噁唑基、喹唑啉基、萘啶基和呋喃并吡啶基等。
本文使用的术语“脂环基”或者“环烷基”是指三个以上碳原子形成的非芳香基环，环内两个相邻的碳原子可以是单键、双键或者三键，环的数目可以是一个或者多个。非限制性的示例包括环丙基、环丁基、环戊基、环己基、环庚基、环己烯基、环戊烯基、环己二烯基等。
本文使用的术语“杂环烷基”或“杂脂环基”是指非芳香基环中一个或多个构成环的原子是选自氮、氧和硫的杂原子。杂环烷基可以由三、四、五、六、七、八、九或多于九个原子构成。杂环烷基可以是任选取代的。杂脂环基的实例包括但不限于内酰胺、内酯、环亚胺、环硫代亚胺、环氨基甲酸酯、四氢噻喃、4H-吡喃、四氢吡喃、哌啶、1,3-二噁英、1,3-二噁烷、1,4-二噁英、1,4-二噁烷、哌嗪、1,3-氧硫杂环己烷、1,4-氧硫杂环己二烯、1,4-氧硫杂环己烷、四氢-1,4-噻嗪、2H-1,2-噁嗪、马来酰亚胺、琥珀酰亚胺、巴比妥酸、硫代巴比妥酸、二氧代哌嗪、乙内酰脲、二氢尿嘧啶、吗啉、三噁烷、六氢-1,3,5-三嗪、四氢噻吩、四氢呋喃、吡咯啉、吡咯烷、咪唑烷、吡咯烷酮、吡唑啉、吡唑烷、咪唑啉、咪唑烷、1,3-二氧杂环戊烯、1,3-二氧杂环戊烷、1,3-二硫杂环戊烯、1,3-二硫杂环戊烷、异噁唑啉、异噁唑烷、噁唑啉、噁唑烷、噁唑烷酮、噻唑啉、噻唑烷和1,3-氧硫杂环戊烷。根据结构，杂脂环基可以是单价基团或双价基团(即亚杂环烷基)。
术语“卤”或“卤素”是指氟、氯、溴和碘。
术语“羰基”是由碳和氧两种原子通过双键连接而成的有机官能团(C＝O)。
术语“任选”指后面描述的一个或多个事件可以或可以不发生，并且包括发生的事件和不发生的事件两者。
本发明中的化合物可能形成的盐也是属于本发明的范围。除非另有说明，本发明中的化合物被理解为 包括其盐类。在此使用的术语“盐”，指用无机或有机酸和碱形成酸式或碱式的盐。此外，当本发明中的化合物含一个碱性片段时，它包括但不限于吡啶或咪唑，含一个酸性片段时，包括但不限于羧酸，可能形成的两性离子(“内盐”)包含在术语“盐”的范围内。药学上可接受的(即无毒，生理可接受的)盐是首选，虽然其他盐类也有用，例如可以用在制备过程中的分离或纯化步骤。本发明的化合物可能形成盐，例如，化合物I与一定量如等当量的酸或碱反应，在介质中盐析出来，或在水溶液中冷冻干燥得来。
本发明中的化合物含有的碱性片段，包括但不限于胺或吡啶或咪唑环，可能会和有机或无机酸形成盐。可以成盐的典型的酸包括醋酸盐(如用醋酸或三卤代醋酸，如三氟乙酸)、己二酸盐、藻朊酸盐、抗坏血酸盐、天冬氨酸盐、苯甲酸盐、苯磺酸盐、硫酸氢盐、硼酸盐、丁酸盐、柠檬酸盐、樟脑盐、樟脑磺酸盐、环戊烷丙酸盐、二甘醇酸盐、十二烷基硫酸盐、乙烷磺酸盐、延胡索酸盐、葡庚糖酸盐、甘油磷酸盐、半硫酸盐、庚酸盐、己酸盐、盐酸盐、氢溴酸盐、氢碘酸盐、羟基乙磺酸盐(如，2-羟基乙磺酸盐)、乳酸盐、马来酸盐、甲磺酸盐、萘磺酸盐(如，2-萘磺酸盐)、烟酸盐、硝酸盐、草酸盐、果胶酸盐、过硫酸盐、苯丙酸盐(如3-苯丙酸盐)、磷酸盐、苦味酸盐、新戊酸盐、丙酸盐，水杨酸盐、琥珀酸盐、硫酸盐(如与硫酸形成的)、磺酸盐、酒石酸盐、硫氰酸盐、甲苯磺酸盐如对甲苯磺酸盐、十二烷酸盐等等。
本发明的某些化合物可能含有的酸性片段，包括但不限于羧酸，可能会和各种有机或无机碱形成盐。典型的碱形成的盐包括铵盐、碱金属盐如钠、锂、钾盐，碱土金属盐如钙、镁盐和有机碱形成的盐(如有机胺)，如苄星、二环已基胺、海巴胺(与N,N-二(去氢枞基)乙二胺形成的盐)、N-甲基-D-葡糖胺、N-甲基-D-葡糖酰胺、叔丁基胺，以及和氨基酸如精氨酸、赖氨酸等等形成的盐。碱性含氮基团可以与卤化物季铵盐，如小分子烷基卤化物(如甲基、乙基、丙基和丁基的氯化物、溴化物及碘化物)，二烷基硫酸盐(如，硫酸二甲酯、二乙酯，二丁酯和二戊酯)，长链卤化物(如癸基、十二烷基、十四烷基和十四烷基的氯化物、溴化物及碘化物)，芳烷基卤化物(如苄基和苯基溴化物)等等。
本发明中化合物的前药及溶剂合物也在涵盖的范围之内。此处术语“前药”是指一种化合物，在治疗相关疾病时，经过代谢或化学过程的化学转化而产生本发明中的化合物、盐、或溶剂合物。“溶剂化物”是指含有化学计量或非化学计量的量的溶剂的溶剂加成形式。一些化合物倾向于在晶体固体状态中捕获固定摩尔比的溶剂分子，从而形成的溶剂化物。如果溶剂是水，形成的溶剂化物是水合物；如果溶剂是醇，形成的溶剂化物是醇化物。一个或多个水分子与一分子的物质的结合形成水合物，其中水保持其分子状态H2O。溶剂化物的非限制性实例包括乙醇溶剂化物、丙酮溶剂化物等。
本发明中的化合物、盐或溶剂合物，可能存在的互变异构形式(例如酰胺和亚胺醚)。所有这些互变异构体都是本发明的一部分。
所有化合物的立体异构体(例如，那些由于对各种取代可能存在的不对称碳原子)，包括其对映体形式和非对映形式，都属于本发明的设想范围。本发明中的化合物独立的立体异构体可能不与其他异构体同时存在(例如，作为一个纯的或者实质上是纯的光学异构体具有特殊的活性)，或者也可能是混合物，如消旋体，或与所有其他立体异构体或其中的一部分形成的混合物。本发明的手性中心有S或R两种构型，由理论与应用化学国际联合会(IUPAC)1974年建议定义。外消旋形式可通过物理方法解决，例如分步结晶，或通过衍生为非对映异构体分离结晶，或通过手性柱色谱法分离。单个的光学异构体可通过合适的方法由外消旋体得到，包括但不限于传统的方法，例如与光学活性酸成盐后再结晶。
本发明中的化合物，依次通过制备、分离纯化获得的该化合物其重量含量等于或大于90％，例如，等于或大于95％，等于或大于99％(“非常纯”的化合物)，在正文描述列出。此处这种“非常纯”本发明的化合物也作为本发明的一部分。
本发明的化合物所有的构型异构体都在涵盖的范围之内，无论是混合物、纯的或非常纯的形式。在本发明化合物的定义包含顺式(Z)和返式(E)两种烯烃异构体，以及碳环和杂环的顺式和反式异构体。
在整个说明书中，基团和取代基可以被选择以提供稳定的片段和化合物。
特定官能团和化学术语定义都详细介绍如下。对本发明来说，化学元素与Periodic Table of the  Elements，CAS version，Handbook of Chemistry and Physics，75th Ed.中定义的一致。特定官能团的定义也在其中描述。此外，有机化学的基本原则以及特定官能团和反应性在“Organic Chemistry”，Thomas Sorrell，University Science Books，Sausalito:1999，也有说明，其全部内容纳入参考文献之列。
本发明的某些化合物可能存在于特定的几何或立体异构体形式。本发明涵盖所有的化合物，包括其顺式和反式异构体、R和S对映异构体、非对映体、(D)型异构体、(L)型异构体、外消旋混合物和其它混合物。另外不对称碳原子可表示取代基，如烷基。所有异构体以及它们的混合物，都包涵在本发明中。
按照本发明，同分异构体的混合物含有异构体的比率可以是多样的。例如，在只有两个异构体的混合物可以有以下组合：50:50，60:40，70:30，80:20，90:10，95:5，96:4，97:3，98:2，99:1，或100:0，异构体的所有比率都在本发明范围之内。本专业内一般技术人员容易理解的类似的比率，及为更复杂的异构体的混合物的比率也在本发明范围之内。
本发明还包括同位素标记的化合物，等同于原始化合物在此公开。不过实际上对一个或更多的原子被与其原子量或质量序数不同的原子取代通常会出现。可以列为本发明的化合物同位素的例子包括氢、碳、氮、氧、磷、硫、氟和氯同位素，分别如2H、3H、13C、11C、14C、15N、18O、17O、31P、32P、35S、18F和36Cl。本发明中的化合物，或对映体，非对映体，异构体，或药学上可接受的盐或溶剂化物，其中含有上述化合物的同位素或其他同位素原子都在本发明的范围之内。本发明中某些同位素标记化合物，例如3H和14C的放射性同位素也在其中，在药物和底物的组织分布实验中是有用的。氚，即3H和碳-14，即14C，它们的制备和检测比较容易。是同位素中的首选。此外，较重同位素取代如氘，即2H，由于其很好的代谢稳定性在某些疗法中有优势，例如在体内增加半衰期或减少用量，因此，在某些情况下可以优先考虑。同位素标记的化合物可以用一般的方法，通过用易得的同位素标记试剂替换为非同位素的试剂，用批露在示例中的方案可以制备。
如果要设计一个本发明的化合物特定的对映体的合成，它可以不对称合成制备，或用手性辅剂衍生化，将所产生的非对映混合物分离，再除去手性辅剂而得到纯的对映体。另外，如果分子中含有一个碱性官能团，如氨基酸，或酸性官能团，如羧基，可以用合适的光学活性的酸或碱的与之形成非对映异构体盐，再通过分离结晶或色谱等常规手段分离，然后就得到了纯的对映体。
如本文所述，本发明中的化合物可与任何数量取代基或官能团取而扩大其包涵范围。通常，术语“取代”不论在术语“可选”前面或后面出现，在本发明配方中包括取代基的通式，是指用指定结构取代基，代替氢自由基。当特定结构中的多个在位置被多个特定的取代基取代时，取代基每一个位置可以是相同或不同。本文中所使用的术语“取代”包括所有允许有机化合物取代。从广义上讲，允许的取代基包括非环状的、环状的、支链的非支链的、碳环的和杂环的，芳环的和非芳环的有机化合物。在本发明中，如杂原子氮可以有氢取代基或任何允许的上文所述的有机化合物来补充其价态。此外，本发明是无意以任何方式限制允许取代有机化合物。本发明认为取代基和可变基团的组合在以稳定化合物形式在疾病的治疗上是很好的。此处术语“稳定”是指具有稳定的化合物，在足够长的时间内检测足以维持化合物结构的完整性，最好是在足够长的时间内都在效，本文在此用于上述目的。
实施例1化合物的合成
化合物RZ001的合成
步骤：向A-1331852(398mg，0.6041mmol，1.00eq)的DMF(8.0ml)溶液里依次加入碳酸钾(160mg，1.92eq)和碘化钾(99mg，0.98eq)，搅拌10分钟后再加入异丁酸氯甲酯(100mg，1.21eq)。油浴外温50℃加热搅拌4h后冷却加水，乙酸乙酯提取，饱和食盐水洗，干燥。浓缩，柱层析。
1HNMR(400MHz,Chloroform-d)δ7.85(m,1H)，7.59(d,1H)，7.42-7.45(m,2H)，7.31-7.34(m,4H)，7.22-7.24(m,1H)，6.93(d,1H)，5.78(s,2H)，5.01(s,2H)，4.03(t,2H)，3.72(s,2H)，3.04(t,2H)，2.51-2.55(m,1H)，2.12(s,3H)，1.95-2.05(m,3H)，1.59-1.72(m,12H)，1.14(s,3H)，1.12(s,3H)
化合物RZ003的合成
步骤：向A-1331852(40mg，0.06071mmol，1.00eq)的DMF(1.0ml)溶液里加入碳酸钾(15mg，1.79eq)，室温搅拌30min后再加入特戊酸氯甲酯(SM7，11mg，1.20eq)的1.0ml DMF溶液。油浴外温50℃加热搅拌5h。乙酸乙酯稀释，水洗，干燥。过滤，浓缩。柱层析纯化。
1HNMR(400MHz,Chloroform-d)δ11.00(s,1H)，7.85(m,1H)，7.54(d,1H)，7.43(m,1H)，7.33(m,1H)，7.28-7.30(m,4H)，7.21-7.23(m,1H)，6.93(d,1H)，5.78(s,2H)，5.00(s,2H)，4.01(t,2H)，3.71(s,2H)，3.03(t,2H)，2.11(s,3H)，1.99-2.05(m,3H)，1.58-1.72(m,12H)，1.16(s,9H)
化合物RZ004的合成
步骤：向A-1331852(50mg，0.07589mmol，1.00eq)的DMF(1.0ml)溶液里加入碳酸钾(17mg，1.62eq)，室温搅拌20min后再加入丁酸氯甲酯(SM8，12mg，1.20eq)的0.8ml DMF溶液。油浴外温50℃加热搅拌4.0h。乙酸乙酯稀释，水洗，干燥。过滤，浓缩，柱层析纯化。
1HNMR(400MHz,Chloroform-d)δ11.20(s,1H)，7.85(m,1H)，7.55(d,1H)，7.43(d,1H)，7.22-7.33(m,5H)，7.17-7.21(m,1H)，6.93(d,1H)，5.77(s,2H)，5.01(s,2H)，4.01(t,2H)，3.71(s,2H)，3.03(t,2H)，2.28(t,2H)，2.10(s,3H)，1.94-1.99(m,3H)，1.57-1.72(m,12H)，1.24-1.33(m,2H)，0.92(t,3H).
化合物RZ005的合成
步骤：向A-1331852(50mg，0.07589mmol，1.00eq)的DMF(1.0ml)溶液里加入碳酸钾(16mg，1.53eq)，室温搅拌30min后再加入乙酸氯甲酯(SM9，10mg，1.21eq)的0.8ml DMF溶液。油浴外温50℃加热搅拌8h后补加乙酸氯甲酯(SM9，8mg，0.97eq)的0.5ml DMF溶液。油浴外温50℃继续加热搅拌3h。乙酸乙酯稀释，水洗，干燥。过滤，浓缩。柱层析纯化。
1HNMR(400MHz,Chloroform-d)δ7.85(m,1H)，7.55(m,2H)，7.44(m,1H)，7.28-7.38(m,5H)，6.93(d,1H)，5.74(s,2H)，5.01(s,2H)，4.03(t,2H)，3.72(s,2H)，3.05(t,2H)，2.12(s,3H)，1.95-2.10(m,6H)，1.55-1.70(m,12H).
化合物RZ012的合成
步骤：向A-1331852(50mg，0.07589mmol，1.00eq)的DMF(1.0ml)溶液里加入碳酸钾(16mg，1.53eq)，室温搅拌30min后再加入苯甲酸氯甲基酯(10mg，1.21eq)的0.8ml DMF溶液。油浴外温50℃继续加热搅拌3h。乙酸乙酯稀释，水洗，干燥。过滤，浓缩。柱层析纯化。
MS(ESI)m/z＝793.5[M+H]+；
依据与上述实施例相同的方法，使用市售化合物或参考所示的中间化合物的制备方法而制备以下表1的实施例化合物。
表1


化合物RZ002的合成
步骤：向A-1331852(30mg，0.04554mmol，1.00eq)的DMF(0.8ml)溶液里加入KHCO3(17mg，3.73eq)，室温搅拌30分钟后再加入氯甲基碳酸异丙酯(8mg，1.20eq)的0.7ml DMF溶液。油浴外温60℃加热搅拌4.5h。乙酸乙酯稀释，水洗，干燥。柱层析纯化。
1HNMR(400MHz,Chloroform-d)δ10.80(s,1H)，7.85(m,1H)，7.55(d,1H)，7.42-7.45(m,2H)，7.30-7.35(m,4H)，7.21-7.24(m,1H)，6.94(d,1H)，5.77(s,2H)，5.00(s,2H)，4.86-4.89(m,1H)，4.03-4.06(m,2H)，3.71(s,2H)，3.04(t,2H)，2.09(s,3H)，1.95-2.05(m,3H)，1.59-1.71(m,12H)，1.24-1.33(m,6H)。
化合物RZ006的合成
步骤：向A-1331852(50mg，0.07589mmol，1.00eq)的DMF(1.0ml)溶液里加入KHCO3(31mg，4.08eq)，室温搅拌20分钟后再加入氯甲基乙基碳酸酯(SM10，13mg，1.24eq)的0.8ml DMF溶液。油浴外温60℃加热搅拌5.0h。乙酸乙酯稀释，水洗，干燥。过滤，浓缩，柱层析纯化。
1HNMR(400MHz,Chloroform-d)δ10.80(s,1H)，7.85(m,1H)，7.55(d,1H)，7.41-7.45(m,2H)，7.29-7.35(m,4H)，7.22-7.24(m,1H)，6.94(d,1H)，5.77(s,2H)，5.00(s,2H)，4.17-4.23(q,2H)，4.02-4.05(m,2H)，3.71(s,2H)，3.04(t,2H)，2.09(s,3H)，1.95-2.05(m,3H)，1.58-1.71(m,12H)，1.24-1.35(m,3H).
依据与上述实施例相同的方法，使用市售化合物或参考所示的中间化合物的制备方法而制备以下表2的实施例化合物。
表2

化合物RZ201的合成
步骤：向A-1331852(50mg，0.07589mmol，1.00eq)的DCM(5.0ml)溶液里加入氯化亚砜(0.1ml)，室温搅拌30min后再加入4-(羟基甲基)-5-甲基-1,3-二氧杂环戊熳-2-酮(SM1,10mg，1.0eq)。室温搅拌3h。二氯甲烷稀释，水洗，干燥，浓缩。柱层析纯化。
MS(ESI)m/z＝771.3[M+H]+；
1H NMR(400MHz,DMSO-d6)δ8.12(s,1H),7.81–7.70(m,2H),7.61(td,J＝8.0,1.2Hz,2H),7.44(dq,J＝8.1,1.1Hz,1H),7.32(dd,J＝7.7,1.7Hz,1H),7.19(td,J＝7.4,1.6Hz,1H),7.06(d,J＝7.9Hz,1H),6.99(td,J＝7.5,1.2Hz,1H),5.05(q,J＝1.1Hz,2H),4.31(d,J＝17.0Hz,1H),4.07(d,J＝17.2Hz,1H),3.96–3.88(m,2H),3.59(s,2H),2.90(ddd,J＝6.8,4.3,1.0Hz,2H),2.71(s,2H),2.08(t,J＝1.0Hz,3H),1.97(pt,J＝5.9,4.2Hz,3H),1.61–1.47(m,13H).
依据与上述实施例相同的方法，使用市售化合物或参考所示的中间化合物的制备方法而制备以下表3的实施例化合物。
表3

化合物RZ301的合成
步骤1：向A-1331852(50mg，0.076mmol，1.00eq)的DMF(1.0ml)溶液里加入碳酸钾(16mg)，室温搅拌30min后再加入N-{[(2-甲基丙-2-基)氧基]羰基}-L-缬氨酸氯甲基酯(24mg，1.2eq)。油浴外温 50℃加热搅拌4h后冷却加水，乙酸乙酯提取，饱和食盐水洗，干燥,浓缩，柱层析得化合物A(50mg,收率：74.1％)。
步骤2：向化合物A(50mg，0.056mmol)的二氧六烷(2ml)溶液里加入4M盐酸/二氧六烷溶液(0.1ml)，室温搅拌1h。二氯甲烷稀释后用饱和碳酸氢钠水溶液调节pH值到7，萃取，干燥，浓缩，柱层析纯化。
MS(ESI)m/z＝788.5[M+H]+；
1H NMR(400MHz,DMSO-d6)δ8.90(d,J＝7.4Hz,2H),7.90(s,1H),7.77(t,J＝8.2Hz,1H),7.73(d,J＝7.9Hz,1H),7.61(td,J＝8.0,1.3Hz,2H),7.44(dq,J＝8.1,1.1Hz,1H),7.32(dd,J＝7.7,1.7Hz,1H),7.19(td,J＝7.4,1.6Hz,1H),7.09–7.03(m,2H),7.03(d,J＝3.3Hz,1H),6.99(td,J＝7.5,1.2Hz,1H),4.31(d,J＝17.0Hz,1H),4.07(d,J＝17.2Hz,1H),3.96–3.88(m,2H),3.55(td,J＝7.2,6.4Hz,1H),2.90(ddd,J＝6.8,4.3,1.0Hz,2H),2.71(s,2H),2.27(dt,J＝13.0,6.5Hz,1H),1.97(pt,J＝5.9,4.2Hz,3H),1.61–1.47(m,15H),1.08(d,J＝6.5Hz,3H),1.02(d,J＝6.5Hz,3H).
实施例2.衰老细胞内的酯酶含量比正常细胞内酯酶含量高
本实施例选取了人非小细胞肺癌细胞系(A549细胞)，人视网膜色素上皮细胞(RPE细胞)，人肺纤维化细胞(IMR90细胞)。检测细胞衰老后其细胞内酯酶含量是否比正常细胞含量更多。从而评估前药在衰老细胞内释放的优势。
将细胞铺于100mm培养皿中，用博来霉素(10μM)和依托泊苷(10μM)诱导衰老。将正常细胞及衰老细胞消化后计数。超声波破碎细胞后离心10min，取上清，置于冰上，按照试剂盒要求进行检测。计算公式：
CarE酶活(U/104cell)＝(ΔA测定管-ΔA空白管)×V反总÷0.5÷(200÷V细胞样总×V样)÷T×F
V样：加入样本体积，0.01mL；T：反应时间，5min；200：细胞总数，200万；V细胞样总：细胞中加入的提取液体积，0.2mL；V反总：反应体系总体积，0.2mL；F：稀释倍数。
从图1A和1B、1C可以看出，衰老细胞内酯酶含量比正常细胞显著升高。
实施例3.前药化合物比A-1331852对衰老细胞具有更特异性的杀伤作用
本实施例选取了人非小细胞肺癌细胞系(A549细胞)，人胚肺成纤维细胞系(IMR90细胞)，人视网膜色素上皮细胞(RPE细胞)，小鼠软骨细胞，小鼠皮肤成纤维细胞。通过依托泊苷诱导(10μM)获得衰老的上述细胞。检测了目标化合物RZ001-RZ006、RZ201对正常及衰老的上述细胞活力的影响，从而评估了化合物前药RZ001-RZ006、RZ201对衰老的上述细胞的特异性杀伤作用。
简单来讲，选取衰老的及正常的上述细胞分别进行铺板(96孔板)。加入化合物RZ001-RZ006、RZ201(浓度为20μM)作用72h后，加入Luminescent Cell Viability Assay试剂盒(Promega)进行细胞活力检测，结果示于图2A～2E中。
从图2A～2E可以看出，A-1331852除了杀伤衰老细胞外，对正常细胞的杀伤作用已十分明显。而化合物RZ001-RZ006、RZ201除了杀伤衰老细胞外，对正常细胞没有明显杀伤。说明RZ001-RZ006、RZ201具有更好的特异性和安全性。
此外，还计算了在正常与衰老A549细胞上的衰老细胞清除指数(Senolytic index)，参见下表4，指数越高说明清除衰老细胞的特异性越强，安全性越好。
表4衰老细胞清除指数




备注：A代表大于200；B代表大于100-200；C代表50-100；D代表30-50；E代表10-30；F代表小
于10。
实施例4：在高氧诱导的视网膜病变的动物模型中的功效
在氧诱导的视网膜病变(OIR)小鼠模型中研究前药RZ001-RZ006以及RZ201的效果，所述模型一定程度指示了早产儿视网膜病变(ROP)、糖尿病性视网膜病变、湿性年龄相关性黄斑变性的体内模型。
自出生后第7天(P7)至第12天(P12)，将C57Bl/6小鼠幼仔和它们的CD1养母暴露于高氧环境(75％O2)。在P12时，对动物玻璃体内注射1μl在1％DMSO，10％吐温80，20％PEG-400中配制的测试组合物(化合物为200uM)，并返回室内空气直至第17天(P17)。在P17时摘除眼球并解剖视网膜用于血管染色。为了测定无血管或新生血管面积，平整视网膜，并用异凝集素B4(IB4)染色。
图3显示玻璃体内(IVT)施用RZ001-RZ006以及RZ201导致无血管区和血管增生的统计学显著改善。
实施例5：在糖尿病诱导的视网膜病变的动物模型中的功效
通过多次施用链唑霉素(STZ)，在糖尿病性视网膜病变的小鼠模型中研究本公开的化合物药效。对6周至7周的C57BL/6J小鼠称重并测量它们的基线血糖(三诺)。以55mg/Kg对小鼠腹膜内注射STZ(Sigma-Alderich)，连续5天。对年龄匹配的对照组仅注射缓冲液。最后一次STZ注射后1周，再次测量血糖。STZ施用后8周和9周对STZ处理的糖尿病C57BL/6J小鼠玻璃体内注射1μl待测化合物(200uM，在0.015％聚山梨酯80，0.2％磷酸钠，0.75％氯化钠，pH7.2中配制成悬浮液)。STZ处理后10周实施视网膜Evans蓝渗透测定法。
图4显示玻璃体内(IVT)施用RZ001，RZ002及RZ201后的视网膜和脉络膜血管渗漏在此剂量水平均可改善血管通透性。
实施例6：在CNV的湿性-年龄相关性黄斑变性的动物模型中的功效
本实施例采用了CNV动物模型，测试了本公开的化合物对湿性-年龄相关性黄斑变性的效果。
雄性小鼠施用0.5％普罗巴卡因盐酸盐(Alcon)用于局部麻醉，并施用1％的托吡卡胺扩张小鼠瞳孔。采用带有裂隙灯输送系统的激光光凝仪(Novus Varia，LUMENIS)进行激光灼烧。而人工撕裂粘附的玻璃盖玻片被用作隐形眼镜。由激光引起的出血斑点不包括在统计数据中。
图5显示玻璃体内(IVT)施用RZ001，RZ002及RZ201后的荧光强度指标的统计学显著改善。
实施例7：在特发性肺纤维化的动物模型中的功效
本实施例采用了特发性肺纤维化动物模型，测试了本公开的的化合物对特发性肺纤维化的效果。
简单来讲，将麻醉好的6周龄的SD大鼠用医用胶带固定在泡沫板上。首先用手术剪刀剪开气管表面的皮肤，充分暴露出气管，将根据大鼠体重吸取的一定体积的博来霉素(BLM 5mg/kg)溶液的100微升的微量进样针，通过口腔伸入到气管中，进行缓慢推注给药。给完BLM溶液后，立即旋转摇晃泡沫板，使BLM药液尽量在肺组织分布均匀。然后用手术缝合针线缝合气管表面的皮肤，并在表皮滴淋大约0.5-1.0ml的青霉素溶液，以防止手术创面的感染。在造模的第7天以后，根据大鼠体重给以本公开的化合物溶液，进行腹腔注射(IP)给药(5mg/kg)，连续给药3周。
在第28天时大鼠进行二氧化碳安乐死，取材肺组织，收集左侧肺泡灌洗液进行白细胞(WBC)计数，右肺组织部分进行羟脯氨酸(HYP)检测，剩余肺组织进行病理的H&E染色和Masson染色。
图6显示腹腔注射(IP)施用RZ001，RZ002和RZ201导致IPF模型上WBC，HYP和病理学评分的统计学显著改善。
实施例8：在骨关节炎的动物模型中的功效
本实施例采用了骨关节炎动物模型，测试了本公开的化合物对于骨关节炎的效果。
通过以下步骤，构建了骨关节炎动物模型。将小鼠采用异氟烷进行麻醉，固定好，剃净小鼠右后肢上的毛发，并用酒精擦拭消毒皮肤，剪开关节部位皮肤。在光学显微镜下，用刀片纵向切开屈膝的右膝关节内测，不要切断髌骨韧带，拉着髌骨韧带到一边，并不断的用无菌棉签进行压迫止血，使得充分暴露出关节腔，并及时滴加生理盐水以免关节腔干，用钝性镊子轻轻分离多余肌肉组织，暴露出半月板韧带，并在关节腔深处寻找前交叉韧带，用微型手术剪切开前交叉韧带，然后用青霉素溶液创面消毒，用灭菌的针线缝合肌肉，最后缝合皮肤组织。通过关节腔内注射给药判断药效。实验终点进行足底继续疼痛阈值检测，取材骨关节进行番红固绿染色。
图7显示关节腔注射施用RZ001，RZ002及RZ201导致骨关节炎(OA)模型上足底机械疼痛阈值和番红固绿染色的病理学评分的统计学显著改善。
实施例9：在对阿尔兹海默症动物模型中的功效
本实施例采用了阿尔兹海默症动物模型，测试了本公开的化合物对阿尔兹海默症的效果。
简单来讲，选择APP×PS1的转基因小鼠模型(采购于赛业生物)，通过腹腔注射给药本公开的化合物，连续给药11周，每周注射2次。观察小鼠大脑皮层淀粉样蛋白的改善。
图8显示腹腔注射施用RZ001，RZ002及RZ201导致AD小鼠模型上Aβ42致病蛋白指标的统计学显著改善。
实施例10：对肝纤维化动物模型中的功效
本实施例采用了肝纤维化动物模型，测试了本公开的化合物对肝纤维化的效果。简单来讲，选用C57BL/6N小鼠，雄性，7-8周龄，体重24-28g。通过腹腔注射CCl4溶液，给药剂量1ml/kg，给药频次为2次/周，造模周期为6周，来诱导小鼠肝纤维化模型。在造模的第三周开始进行腹腔注射给药(5mg/kg)，每周给药两次。实验终点采集血清，取材肝组织，进行Masson染色指标检测。
图9显示腹腔注射(IP)施用RZ001，RZ002和RZ201导致肝纤维化模型上血清中谷草转氨酶(AST)和谷丙转氨酶(ALT)水平和病理学的统计学显著改善。
实施例11：对衰老皮肤的功效
本实施例采用了老年雌性C57bl/6J小鼠(22月龄)。通过皮肤涂抹给药。每周给药两次，连续给药6周。观察老年小鼠皮肤表皮厚度的改善情况。
图10显示涂抹施用RZ001，RZ002和RZ201导致自然衰老小鼠模型皮肤表皮厚度评分的统计学显著改善。
实施例12：对肾纤维化动物模型中的功效
本实施例采用了肾纤维化动物模型，测试了本公开的化合物对肾纤维化的效果。简单来讲，选用C57BL/6J小鼠，雄性，7-8周龄。将小鼠麻醉后，使小鼠仰卧于泡沫板上，碘伏消毒，在下腹部正中皮肤纵向切口，找到腹肌白线，在此处剖开腹肌，筋膜和腹膜层。暴露手术视野后，用浸有生理盐水的纱布将小鼠的胃及肠系膜推向右侧暴露肾脏。从肾盂处方沿着肾动脉向下找到透明状的输尿管，分离左侧输尿管，在左侧输尿管上1/3处结扎，逐层缝合切口，最后用碘伏消毒，以此来构建肾纤维化模型。在造模的第2周开始进行腹腔注射给药。实验终点采集小鼠血清，取材肾组织进行Masson染色指标检测。
图11显示腹腔注射(IP)施用RZ001，RZ002和RZ201导致肾纤维化模型上血清中肌酐(Cr)和尿素氮(BUN)水平和病理学的统计学显著改善。
实施例13：对兔耳增生性瘢痕模型中的功效
本实施例采用了兔耳增生性瘢痕动物模型，测试了本公开的化合物对增生性瘢痕的效果。简单来讲，选用10只新西兰大白兔，雄性，3月龄。将新西兰兔用戊巴比妥钠麻醉后，通过去除表皮、真皮和软骨膜至裸露的软骨，在每只耳朵的腹侧表面制作6个直径10mm的圆形全层伤口。兔随机分为对照组和RZ001实验组。在术后第14天，创面完全重新上皮化后，取化合物(150uM,100ul，溶于DMSO溶液)或阴性对照DMSO 100ul，从伤口边缘将溶液注射到每个病变的中心，每周注射一次，共注射4次。通过SEI(瘢痕增生指数)为评估化合物瘢痕形成的改善效果。
图12显示施用RZ001导致兔耳增生性瘢痕模型瘢痕增生指数统计学显著改善。

Claims (15)

  1. 一种化合物，其具有式I所示的结构，或其药学上可接受的盐、溶剂化物、水合物、多晶型物、共晶、互变异构体、立体异构体或同位素标记化合物，
    其中，
    R1选自取代或未取代的C1-C10亚烷基；R2选自-O-C(O)-或者-O-C(O)-O-；
    R3选自取代或未取代的C1-C10链烷基、取代或未取代的C3-C20环烷基、取代或未取代的C2-C20链烯基、取代或未取代的C2-C20链炔基、取代或未取代的C6-C20芳基、取代或未取代的C1-C20杂芳基、取代或未取代的C2-C20杂脂环基；
    或者，R1不存在或者选自取代或未取代的C1-C10亚烷基，R2不存在，并且R3选自取代或未取代的C2-C20杂脂环基；
    Rc选自取代或未取代的金刚烷基；
    可选地，所述取代中的取代基选自卤素原子、氰基、硝基、C6-C20芳基、C1-C20杂芳基、C1-C10链烷基、C1-C10链烷氧基、C6-C20芳氧基、C1-C20杂脂环基优选C1-C10杂脂环基、氨基、羟基、巯基、磷酸酯基、-OC(O)R6、-ONR6R7、-NR6R7的取代基取代，R6和R7独立选自氢、C6-C20芳基、C1-C20杂芳基、C1-C8链烷基、C3-C8环烷基、C2-C8链烯基和C2-C8链炔基，所述芳基和杂芳基任选被卤素原子、羟基、巯基、氨基、硝基、氰基、羧基、酰基、C1-C6烷氧基、C6-C20芳基、C1-C20杂芳基、C2-C20杂脂环基、C1-C10烷基、C3-C8环烷基、C2-C8链烯基或C2-C8链炔基取代，其中任选至少两个位置的取代基共同构成脂肪环例如C3-C20脂肪环、杂脂环例如C2-C20脂肪环、芳环例如C6-C20芳环或杂芳环例如C1-C20杂芳环。
  2. 根据权利要求1所述的化合物，其特征在于，所述化合物如式III、IV或V所示：

    式III和IV中，R1选自独立选自取代或未取代的C1-C10亚烷基，优选地，R1选自独立选自取代或未取代的C1-C6亚烷基，优选地，R1选自独立选自取代或未取代的C1-C4亚烷基；Ra、Rb、R3、Rc、L的定义同式I；
    式V中，R1不存在或者选自独立选自取代或未取代的C1-C10亚烷基，优选地，R1选自独立选自取代或未取代的C1-C6亚烷基，优选地，R1选自独立选自取代或未取代的C1-C4亚烷基，R3选自取代或未取代的C2-C20杂脂环基，Ra、Rb、Rc、L的定义同式I。
  3. 根据权利要求1或2所述的化合物，其特征在于，
    所述C2-C20杂脂环基任选地被选自卤素原子、羟基、巯基、氨基、硝基、氰基、C1-C10烷氧基、C1-C10烷基、C3-C8环烷基、C2-C8链烯基、C2-C8炔基、 的取代基取代，各R5独立选自氢和C1-C6烷基。
  4. 根据权利要求1或2所述的化合物，其特征在于，所述C2-C20杂脂环基是C4-C8杂脂环基；
    所述C4-C8杂脂环基任选地被选自卤素、-NH2、-OH、-NO2、羰基、-CH2OH、羧基、甲基、乙基、丙基、异丙基、甲氧基、乙氧基、丙氧基或异丙氧基取代。
  5. 根据权利要求1-4中任一项所述的化合物，其特征在于，所述C2-C20杂脂环基选自下组所示基团：

    R’各自独立地表示无取代基、单个取代基或多个取代基，各取代基独立地选自氘、羟基、卤素、NH2、羧基(-COOH)、C1-C6链烷基、卤素取代的C1-C6链烷基、羟基取代的C1-C6链烷基、氨基取代的C1-C6链烷基、吗啉取代的C1-C6链烷基、-COO-C1-C6链烷基、氰基、C1-C6链烷氧基、C3-C6环烷基、卤素取代的C3-C6环烷基、羟基取代的C3-C6环烷基、苯基、苄基；
    L2为不存在C1-C6亚烷基、卤素、羟基、C1-C6烷氧基取代的C1-C6亚烷基，优选亚甲基、亚乙基、亚丙基；
    R6为H、氘、卤素、羟基、NH2、羧基(-COOH)、-CONH2、磺酸基(-SO3H)、磺酰基-(SO2)-C1-C6链烷基、C1-C6链烷基、卤素取代的C1-C6链烷基、羟基取代的C1-C6链烷基、氨基取代的C1-C6链烷基、C3-C6环烷基、卤素取代的C3-C6环烷基、羟基取代的C3-C6环烷基、苯基或苄基。
  6. 根据权利要求1-5任一项所述的化合物，其特征在于，各Ra均为氢；和/或
    Rb选自取代或未取代的C1-C6烷基，优选取代或未取代的C1-C3烷基，更优选甲基；和/或
    L选自取代或未取代的C1-C6亚烷基，优选取代或未取代的C1-C3亚烷基，更优选亚甲基。
  7. 根据权利要求1-6任一项所述的化合物，其特征在于，所述化合物如式II所示：
    式II中，R1、R2、R3的限定同式I。
  8. 根据权利要求1-7任一项所述的化合物，其特征在于，R1为取代或未取代的C1-C6亚烷基，优选为取代或未取代的C1-C3亚烷基，更优选为亚甲基，和/或
    R3选自取代或未取代的C1-C8链烷基、取代或未取代的C3-C10环烷基、取代或未取代的C2-C8链烯基、取代或未取代的C2-C8链炔基、取代或未取代的C6-C12芳基、取代或未取代的C2-C12杂芳基或者取代或未取代的C1-C12杂脂环基；
    优选地，R3选自取代或未取代的C1-C6烷基，取代或未取代的C6-C12芳基，取代或未取代的C1-C10杂芳基，取代或未取代的C3-C8环烷基，或者取代或未取代的C1-C10杂脂环基。
  9. 根据权利要求1-8任一项所述的化合物，其特征在于，所述取代为被选自卤素原子、氰基、硝基、C6-C20芳基、C1-C20杂芳基、C1-C10链烷基、C1-C10链烷氧基、C6-C20芳氧基、C1-C20杂脂环基优选C1-C10杂脂环基、氨基、羟基、巯基、磷酸酯基、-OC(O)R6、-ONR6R7、-NR6R7的取代基取代，R6和R7独立选自氢、C6-C20芳基、C1-C20杂芳基、C1-C8链烷基、C3-C8环烷基、C2-C8链烯基和C2-C8链炔基，所述芳基和杂芳基任选被卤素原子、羟基、巯基、氨基、硝基、氰基、羧基、酰基、C1-C6烷氧基、C6-C20芳基、C1-C20杂芳基、C2-C20杂脂环基、C1-C10烷基、C3-C8环烷基、C2-C8链烯基或C2-C8链炔基取代，其中任选至少两个位置的取代基共同构成脂肪环例如C3-C20脂肪环、杂脂环例如C2-C20脂肪环、芳环例如C6-C20芳环或杂芳环例如C1-C20杂芳环。
  10. 根据权利要求1-9任一项所述的化合物，其特征在于，R3选自以下基团：
  11. 根据权利要求1所述的化合物，其特征在于，所述化合物选自以下化合物：




  12. 一种用于预防或治疗与衰老相关疾病的方法，包括向有需要的受试者施用治疗有效量的权利要求1至10中任一项所述的化合物或其药学上可接受的盐、溶剂化物、水合物、多晶型物、共晶、互变异构体、立体异构体或同位素标记化合物。
  13. 根据权利要求12所述的方法，其特征在于，与衰老相关疾病是衰老细胞的积累相关的疾病，优选地，所述疾病选自特发性肺纤维化、肺纤维化、肝纤维化、肾纤维化、病毒导致的上呼吸道和肺部的炎症和组织纤维化及萎缩、囊性纤维化、骨髓纤维化、心肌纤维化、皮肤纤维化、间质性肺疾病、纤维化的胰腺炎、早产儿视网膜病变、黄斑变性、糖尿病性黄斑水肿、糖尿病性视网膜病变、年龄相关性黄斑变性、湿性年龄相关黄斑变性、干性年龄相关黄斑变性、青光眼、镰状细胞视网膜病变、缺血性动脉炎性神经病变、干燥性角膜炎、Fuch氏角膜营养不良、老视、白内障、变性玻璃体病症，包括玻璃体黄斑牵引综合征、黄斑裂孔、视网膜撕裂、视网膜剥离、增殖性玻璃体视网膜病变、骨关节炎、椎间盘突出、骨质疏松、阿尔兹海默症、帕金森病、动脉粥样硬化、慢性阻塞性肺病、糖尿病、糖尿病肾病、瘢痕、表浅性瘢痕或扁平瘢痕、索条状瘢痕或挛缩性瘢痕、蹼状瘢痕、凹陷性瘢痕、萎缩性瘢痕、桥状瘢痕和赘状瘢痕、增生性瘢痕、瘢痕疙瘩、瘢痕癌、硬皮病、硬斑病、带状硬皮病、点滴状硬皮病、肢端硬皮病、弥漫性硬皮病、CREST综合征、急性冠状动脉综合征、心肌梗塞、中风、高血压、肥胖、脂肪功能障碍、冠状动脉疾病、脑血管疾病、牙周病、癌症治疗相关的残疾例如各种组织中的萎缩和纤维化、脑和心脏损及治疗相关的骨髓增生异常综合征、早幼粒综合征、共济失调毛细血管扩张症、范可尼贫血、弗里德赖希共济失调、先天性角化不良、再生障碍性贫血、动脉瘤、炎症性肠病、脂肪萎缩、肾脏移植手术失败、肌肉衰减征、伤口愈合、秃头症、心肌细胞肥大、肾小球硬化症、和癌症中的一种或多种。
  14. 权利要求1至11中任一项所述的化合物或其药学上可接受的盐、溶剂化物、水合物、多晶型物、共晶、互变异构体、立体异构体或同位素标记化合物在制备用于防或治疗与衰老相关疾病的药物中的应用。
  15. 根据权利要求14所述的应用，其特征在于，与衰老相关疾病是衰老细胞的积累相关的疾病，优选地，所述疾病选自衰老细胞的积累相关的疾病，所述疾病优选选自特发性肺纤维化、肺纤维化、肝纤维化、 肾纤维化、病毒导致的上呼吸道和肺部的炎症和组织纤维化及萎缩、囊性纤维化、骨髓纤维化、心肌纤维化、皮肤纤维化、间质性肺疾病、纤维化的胰腺炎、早产儿视网膜病变、黄斑变性、糖尿病性黄斑水肿、糖尿病性视网膜病变、年龄相关性黄斑变性、湿性年龄相关黄斑变性、干性年龄相关黄斑变性、青光眼、镰状细胞视网膜病变、缺血性动脉炎性神经病变、干燥性角膜炎、Fuch氏角膜营养不良、老视、白内障、变性玻璃体病症，包括玻璃体黄斑牵引综合征、黄斑裂孔、视网膜撕裂、视网膜剥离、增殖性玻璃体视网膜病变、骨关节炎、椎间盘突出、骨质疏松、阿尔兹海默症、帕金森病、动脉粥样硬化、慢性阻塞性肺病、糖尿病、糖尿病肾病、瘢痕、表浅性瘢痕或扁平瘢痕、索条状瘢痕或挛缩性瘢痕、蹼状瘢痕、凹陷性瘢痕、萎缩性瘢痕、桥状瘢痕和赘状瘢痕、增生性瘢痕、瘢痕疙瘩、瘢痕癌、硬皮病、硬斑病、带状硬皮病、点滴状硬皮病、肢端硬皮病、弥漫性硬皮病、CREST综合征、急性冠状动脉综合征、心肌梗塞、中风、高血压、肥胖、脂肪功能障碍、冠状动脉疾病、脑血管疾病、牙周病、癌症治疗相关的残疾例如各种组织中的萎缩和纤维化、脑和心脏损及治疗相关的骨髓增生异常综合征、早幼粒综合征、共济失调毛细血管扩张症、范可尼贫血、弗里德赖希共济失调、先天性角化不良、再生障碍性贫血、动脉瘤、炎症性肠病、脂肪萎缩、肾脏移植手术失败、肌肉衰减征、伤口愈合、秃头症、心肌细胞肥大、肾小球硬化症、和癌症中的一种或多种。
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