WO2022017412A1 - 治疗造血干细胞移植后的移植物抗宿主病的方法 - Google Patents

治疗造血干细胞移植后的移植物抗宿主病的方法 Download PDF

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WO2022017412A1
WO2022017412A1 PCT/CN2021/107575 CN2021107575W WO2022017412A1 WO 2022017412 A1 WO2022017412 A1 WO 2022017412A1 CN 2021107575 W CN2021107575 W CN 2021107575W WO 2022017412 A1 WO2022017412 A1 WO 2022017412A1
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compound
formula
pharmaceutically acceptable
alkyl
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PCT/CN2021/107575
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English (en)
French (fr)
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郑翔玲
赵焰平
王红军
仲伟婷
李晶
赵静
刘伟娜
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北京泰德制药股份有限公司
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Priority to KR1020237005450A priority Critical patent/KR20230043891A/ko
Priority to CA3186564A priority patent/CA3186564A1/en
Priority to BR112023001049A priority patent/BR112023001049A2/pt
Priority to JP2023504356A priority patent/JP2023535035A/ja
Priority to CN202180059344.8A priority patent/CN116322670A/zh
Priority to IL299985A priority patent/IL299985A/en
Priority to EP21845732.3A priority patent/EP4186508A1/en
Priority to US18/017,198 priority patent/US20230285393A1/en
Priority to AU2021313124A priority patent/AU2021313124A1/en
Publication of WO2022017412A1 publication Critical patent/WO2022017412A1/zh

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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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A61K31/416—1,2-Diazoles condensed with carbocyclic ring systems, e.g. indazole
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings

Definitions

  • the present invention belongs to the field of biomedicine, and specifically relates to a method for preventing, alleviating and/or treating graft-versus-host disease after hematopoietic stem cell transplantation, which comprises administering an effective amount of a compound of the present application or its pharmacy to an individual in need thereof.
  • hematopoietic tumors such as leukemia
  • hematopoietic stem cell eg peripheral blood stem cells, bone marrow cells, etc.
  • transplantation is required for patients who are refractory to or less likely to be cured by conventional chemotherapy.
  • graft-versus-host disease GVHD
  • aGVHD acute graft-versus-host disease
  • cGVHD chronic transplantation Object-versus-host disease
  • aGVHD mainly involves the skin, gastrointestinal tract, and liver, and in rare cases other organs may also be involved. Skin damage alone is usually not life-threatening; if visceral damage occurs, severe jaundice, intractable diarrhea and bloody stools, colic, and severe systemic symptoms occur. Liver or gastrointestinal impairment occurs in up to 60% of all patients with aGVHD, and mortality is 85%.
  • cGVHD develops after transplantation and can involve multiple organs, such as the skin, mouth, eyes, gastrointestinal tract, liver, lungs, joints, fascia, and reproductive tract.
  • the present invention provides a method of preventing, ameliorating and/or treating graft-versus-host disease following hematopoietic stem cell transplantation, comprising administering to an individual in need thereof an effective amount of a compound of formula (I) or a pharmaceutically acceptable agent thereof Acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs:
  • Ring A is The above group is attached to the pyrimidine ring through one of the two positions marked with * or **, and the other position is attached to the carbonyl group;
  • R is selected from H and C 1-6 alkyl
  • R 2 is selected from H and C 1-6 alkyl
  • R 3 , R 4 , R 7 and R 8 at each occurrence are each independently selected from H, halogen, -NR 5 R 6 , -OH, C 1-6 alkyl and -OR 5 ;
  • alkylene, alkyl, alkenyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and arylalkyl are each optionally substituted at each occurrence with one or more substituents independently selected from halogen, C 1 Substituent substitution of -6 alkyl and -OR 5;
  • R 5 and R 6 at each occurrence are each independently selected from H, C 1-6 alkyl, C 3-10 cyclohydrocarbyl, 3-10 membered heterocyclyl, C 6-10 aryl, 5-14 membered Heteroaryl and C 6-12 aralkyl;
  • n is independently at each occurrence an integer of 0, 1, 2 or 3;
  • n is independently an integer of 0, 1 or 2.
  • the present invention provides compounds of formula (I) above, or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically-labeled compounds thereof , use of a metabolite or a prodrug in the preparation of a medicament for preventing, relieving and/or treating graft-versus-host disease after hematopoietic stem cell transplantation.
  • the present invention provides compounds of formula (I) above, or pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically-labeled compounds thereof , metabolites or prodrugs for the prevention, mitigation and/or treatment of graft-versus-host disease after hematopoietic stem cell transplantation.
  • FIG. 1 shows the survival rate of mice in Example 2.
  • Figure 2 shows the inhibitory effect of compound 007 tested in Example 3 on inflammatory factors (*P ⁇ 0.05, ***P ⁇ 0.001, ****P ⁇ 0.0001 compared to the control group).
  • alkylene refers to a saturated divalent hydrocarbon radical, preferably a saturated divalent hydrocarbon radical having 1, 2, 3, 4, 5 or 6 carbon atoms, such as methylene, ethylene, propylene or butylene.
  • alkyl is defined as a linear or branched saturated aliphatic hydrocarbon.
  • the alkyl group has 1 to 12, eg, 1 to 6, carbon atoms.
  • C 1-6 alkyl refers to a linear or branched group of 1 to 6 carbon atoms (eg, methyl, ethyl, n-propyl, isopropyl, n-butyl) , isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl or n-hexyl) optionally substituted by 1 or more (such as 1 to 3) suitable substituents the halo-substituted (in this case the group is referred to as "haloalkyl”) (e.g.
  • C 1-4 alkyl refers to a linear or branched aliphatic hydrocarbon chain of 1 to 4 carbon atoms (ie, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl).
  • alkenyl means a linear or branched monovalent hydrocarbon group containing one double bond and having 2-6 carbon atoms (“C 2-6 alkenyl”).
  • the alkenyl groups are, for example, vinyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2- - Hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl and 4-methyl-3-pentenyl.
  • the compound of the present invention contains an alkenylene group, the compound may exist in pure E (ent ought) form, pure Z (zusammen) form, or any mixture thereof.
  • alkynyl refers to a monovalent hydrocarbon group containing one or more triple bonds, preferably having 2, 3, 4, 5 or 6 carbon atoms, such as ethynyl or propynyl.
  • cycloalkyl refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (eg, monocyclic such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl) , cyclooctyl, cyclononyl, or bicyclic, including spiro, fused, or bridged systems (such as bicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl or bicyclo[5.2.0]nonyl, decalinyl, etc.)), which are optionally substituted with 1 or more (such as 1 to 3) suitable substituents.
  • monocyclic such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl
  • cyclooctyl cyclonony
  • the cycloalkyl group has 3 to 15 carbon atoms.
  • C 3-6 cycloalkyl refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (eg, cyclopropyl, cyclobutyl, cyclopentyl or cyclo) of 3 to 6 ring carbon atoms hexyl), which is optionally substituted with 1 or more (such as 1 to 3) suitable substituents, eg methyl substituted cyclopropyl.
  • cyclohydrocarbylene refers to rings having, for example, 3-10 (suitably 3-8, more suitably 3-6) ring carbons Atomically saturated (ie, “cycloalkylene” and “cycloalkyl”) or unsaturated (ie, having one or more double and/or triple bonds in the ring) monocyclic or polycyclic hydrocarbon rings, which Including but not limited to ()cyclopropylidene (ring), ()cyclobutylidene (ring), ()cyclopentylidene (ring), ()cyclohexylene (ring), ()cycloheptidene ( cyclo), () cyclooctyl (ring), () cyclononyl (ring), () cyclohexenyl (ring), and the like.
  • heterocyclyl As used herein, the terms “heterocyclyl”, “heterocyclylene” and “heterocycle” mean having, for example, 3-10 (suitably 3-8, more suitably 3-6) Ring atoms in which at least one ring atom is a heteroatom selected from N, O, and S and the remaining ring atoms are saturated (ie, heterocycloalkyl) or partially unsaturated (ie, have one or more within the ring double and/or triple bonds) cyclic groups.
  • a "3-10 membered (sub)heterocycle (radical)" is one having 2-9 (eg, 2, 3, 4, 5, 6, 7, 8, or 9) ring carbon atoms and is independently selected from N A saturated or partially unsaturated (sub)heterocycle (radical) of one or more (eg 1, 2, 3 or 4) heteroatoms of , O and S.
  • heterocyclylenes and heterocycle(radicals) include, but are not limited to: ()oxiranyl, ()aziridinyl, (azetidinyl), ()oxygenide Heterocyclobutyl (oxetanyl), ()tetrahydrofuranyl, ()dioxolinyl (dioxolinyl), ()pyrrolidine, ()pyrrolidone, ()imidazolidinylene, () ) Pyrazolidine, () Pyrrolidene, () Tetrahydropyranyl, () Piperidinyl, () Morpholinyl, () Dithianyl (dithianyl), () Thiomorpholinyl, ()piperazinylidene or (trithianylidene)trithianyl.
  • the groups also encompass bicyclic ring systems, including spiro, fused or bridged systems (such as 8-azaspiro[4.5]decane, 3,9-diazaspiro[5.5]undecane, 2-nitrogen Heterobicyclo[2.2.2]octane, etc.).
  • Heterocyclylene and heterocycle(radicals) may be optionally substituted with one or more (eg, 1, 2, 3, or 4) suitable substituents.
  • the terms "()arylene” and “aromatic ring” refer to an all-carbon monocyclic or fused ring polycyclic aromatic group having a conjugated pi electron system.
  • C 6-10 ()arylene” and “C 6-10 aromatic ring” mean an aromatic group containing 6 to 10 carbon atoms, such as ()phenylene (benzene ring) or ()naphthylene (naphthalene ring).
  • (Alkylene) aryl and aryl ring is optionally substituted with one or more (such as 1 to 3) suitable substituent (e.g., halo, -OH, -CN, -NO 2, C 1-6 alkyl, etc.) substituted with .
  • suitable substituent e.g., halo, -OH, -CN, -NO 2, C 1-6 alkyl, etc.
  • heteroarylidene and heteroaryl ring refer to monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 1 or 2 or 3 or 4 or 5 or 6 or 9 or 10 carbon atoms, and which contain at least one heteroatom (such as oxygen, nitrogen, etc.) which may be the same or different or sulfur) and, in addition, can be benzo-fused in each case.
  • heteroatom such as oxygen, nitrogen, etc.
  • "()heteroarylene” or “heteroaromatic ring” is selected from ()thienylene, ()furanyl, ()pyrrolylene, ()oxazolylylene, ()thiazolylylene, ()imidazolylidene, ()pyrazolylidene, ()isoxazolylidene, ()isothiazolylidene, ()oxadiazolylidene, ()triazolylidene, ()thiadiazolylidene etc., and their benzo derivatives; or ()pyridylene, ()pyridazinylene, ()pyrimidinylene, ()pyrazinylene, ()triazinylene, etc., and their benzos derivative.
  • aralkyl preferably refers to an aryl or heteroaryl substituted alkyl group, wherein said aryl, heteroaryl and alkyl groups are as defined herein.
  • the aryl group can have 6-14 carbon atoms
  • the heteroaryl group can have 5-14 ring atoms
  • the alkyl group can have 1-6 carbon atoms.
  • Exemplary aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, phenylbutyl.
  • halo or halogen group is defined to include F, Cl, Br or I.
  • substituted means that one or more (eg, one, two, three, or four) hydrogens on the designated atom are replaced by a selection from the designated group, provided that no more than the designated atom is present in the normal valences in the case and the substitutions form stable compounds. Combinations of substituents and/or variables are permissible only if such combinations form stable compounds.
  • substituent can be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more of the list of substituents, one or more hydrogens on the carbon (to the extent of any hydrogens present) may be independently and/or together independently Selected optional substituents are substituted. If a nitrogen of a substituent is described as being optionally substituted with one or more of the list of substituents, then one or more hydrogens on the nitrogen (to the extent of any hydrogens present) may each be independently selected optional substitution of substituents.
  • each substituent is selected independently of the other.
  • each substituent may be the same as or different from another (other) substituent.
  • one or more means 1 or more than 1, such as 2, 3, 4, 5 or 10, under reasonable conditions.
  • the point of attachment of a substituent can be from any suitable position on the substituent.
  • the present invention also includes all pharmaceutically acceptable isotopically-labeled compounds that are identical to the compounds of the present invention, except that one or more atoms have the same atomic number but an atomic mass or mass number different from the atomic mass that predominates in nature or atomic substitution of mass number.
  • suitable contain a compound of the present invention isotopes include (but are not limited to) isotopes of hydrogen (e.g., deuterium (2 H), tritium (3 H)); isotopes of carbon (e.g.
  • isotopes of chlorine eg 36 Cl
  • isotopes of fluorine eg 18 F
  • isotopes of iodine eg 123 I and 125 I
  • isotopes of nitrogen eg 13 N and 15 N
  • isotopes of oxygen eg 15 O
  • isotopes of phosphorus eg 32 P
  • isotopes of sulfur eg 35 S.
  • Certain isotopically-labeled compounds of the invention eg, those incorporating radioisotopes
  • are useful in drug and/or substrate tissue distribution studies eg, assays).
  • radioisotopes tritium (ie 3 H) and carbon-14 (ie 14 C) are particularly useful for this purpose due to their ease of incorporation and ease of detection.
  • Substitution with positron emitting isotopes e.g., 11 C, 18 F, 15 O and 13 N
  • PET emission tomography positron
  • Isotopically labeled compounds of the invention can be prepared by methods analogous to those described in the accompanying Schemes and/or Examples and Preparations by using an appropriate isotopically labeled reagent in place of the previously employed non-labeled reagent.
  • Pharmaceutically acceptable solvates of the present invention include those wherein the solvent of crystallization may be isotopically substituted, e.g., D 2 O, acetone -d 6 or DMSO-d 6.
  • stereoisomer refers to isomers formed due to at least one asymmetric center. In compounds having one or more (eg, one, two, three or four) asymmetric centers, it may give rise to racemic mixtures, single enantiomers, diastereomeric mixtures and individual of diastereomers. Certain individual molecules can also exist as geometric isomers (cis/trans). Similarly, the compounds of the present invention may exist as mixtures of two or more structurally distinct forms in rapid equilibrium (often referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers Wait. It is to be understood that the scope of this application covers all such in any ratio (eg 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% %) of isomers or mixtures thereof.
  • Solid lines may be used in this article solid wedge or virtual wedge
  • the chemical bonds of the compounds of the present invention are depicted.
  • the use of a solid line to depict a bond to an asymmetric carbon atom is intended to indicate that all possible stereoisomers at that carbon atom are included (eg, a specific enantiomer, racemic mixture, etc.).
  • the use of real or dashed wedges to delineate bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomer exists.
  • real and imaginary wedges are used to define relative, rather than absolute, stereochemistry.
  • the compounds of the present invention are intended to be available as stereoisomers (which include cis and trans isomers, optical isomers (eg, R and S enantiomers), diastereomers, Geometric isomers, rotational isomers, conformational isomers, atropisomers and mixtures thereof).
  • stereoisomers which include cis and trans isomers, optical isomers (eg, R and S enantiomers), diastereomers, Geometric isomers, rotational isomers, conformational isomers, atropisomers and mixtures thereof).
  • the compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (eg, racemic mixtures and pairs of diastereomers).
  • the present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be a single polymorph or a mixture of more than one polymorph in any ratio.
  • compositions of the present invention may exist in free form for use in therapy, or, where appropriate, in the form of their pharmaceutically acceptable derivatives.
  • pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, N-oxides, metabolites or prodrugs which are administered to patients in need thereof After administration, the compounds of the invention or their metabolites or residues can be provided directly or indirectly. Accordingly, references herein to "compounds of the present invention" are also intended to encompass the various derivative forms of the compounds described above.
  • Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof.
  • Suitable acid addition salts are formed from acids which form pharmaceutically acceptable salts. Examples include acetate, adipate, aspartate, benzoate, besylate, bicarbonate/carbonate, bisulphate/sulphate, borate, camphorsulphonate , citrate, cyclamate, ethanedisulfonate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate Salt, Hyphenate, Hydrochloride/Chloride, Hydrobromide/Bromide, Hydroiodide/Iodide, Isethionate, Lactate, Malate, Maleic Acid salt, malonate, mesylate, methyl sulfate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitic acid Salt, Pamoate, Phosphat
  • Suitable base addition salts are formed from bases which form pharmaceutically acceptable salts. Examples include aluminum salts, arginine salts, benzathine penicillin salts, calcium salts, choline salts, diethylamine salts, diethanolamine salts, glycinate salts, lysine salts, magnesium salts, meglumine salts, ethanolamine salts, Potassium, sodium, tromethamine and zinc salts.
  • esters means an ester derived from each of the compounds of the general formula in this application, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release free acid or alcohol forms of the present invention) compound).
  • the compounds of the present invention may themselves also be esters.
  • the compounds of the present invention may exist in the form of solvates, preferably hydrates, wherein the compounds of the present invention comprise a polar solvent as a structural element of the crystal lattice of the compound, in particular for example water, methanol or ethanol.
  • a polar solvent as a structural element of the crystal lattice of the compound, in particular for example water, methanol or ethanol.
  • the amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.
  • Nitrogen-containing heterocycles are capable of forming N-oxides since nitrogen requires available lone pairs of electrons to oxidize to oxides; Nitrogen-containing heterocycles. Those skilled in the art will also recognize that tertiary amines are capable of forming N-oxides.
  • N-oxides of heterocycles and tertiary amines are well known to those skilled in the art and include the use of peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (MCPBA), hydrogen peroxide, alkyl Hydrogen peroxides such as t-butyl hydroperoxide, sodium perborate and dioxiranes such as dimethyldioxirane are used to oxidize heterocycles and tertiary amines.
  • peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (MCPBA)
  • hydrogen peroxide alkyl Hydrogen peroxides such as t-butyl hydroperoxide
  • sodium perborate and dioxiranes such as dimethyldioxirane
  • metabolites of the compounds of the present invention ie substances formed in the body upon administration of the compounds of the present invention. Such products may result from, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, and the like, of the administered compound.
  • the present invention includes metabolites of the compounds of the present invention, including compounds prepared by methods of contacting a compound of the present invention with a mammal for a time sufficient to produce the metabolites thereof.
  • the present invention further includes within its scope prodrugs of the compounds of the present invention, which are certain derivatives of the compounds of the present invention that may themselves have little or no pharmacological activity when administered into or onto the body can be converted into compounds of the invention having the desired activity, for example, by hydrolytic cleavage.
  • prodrugs will be functional derivatives of the compound that are readily converted in vivo to the desired therapeutically active compound. Additional information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems", Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella).
  • prodrugs of the present invention can be obtained, for example, by using certain moieties known to those skilled in the art as “pro-moiety (eg as described in “Design of Prodrugs", H. Bundgaard (Elsevier, 1985))" Prepared by substituting appropriate functional groups present in the compounds of the present invention.
  • the present invention also encompasses compounds of the present invention that contain protecting groups.
  • protecting groups In any process for preparing the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any relevant molecule, thereby forming chemically protected forms of the compounds of the present invention. This can be accomplished by conventional protecting groups, such as those described in T.W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.
  • an effective amount refers to an amount sufficient to achieve the desired therapeutic effect under the conditions of administration, which results in pathological symptoms, disease progression, amelioration of physiological conditions associated therewith, or induction of resistance to the aforementioned diseases.
  • treating means reversing, alleviating, inhibiting the progression of the disorder or condition to which such term applies or one or more symptoms of such disorder or condition, or Such a disorder or condition or one or more symptoms of such a disorder or condition is prevented.
  • an “individual” as used herein includes a human or non-human animal.
  • exemplary human subjects include human subjects (referred to as patients) or normal subjects with a disease (eg, a disease described herein).
  • Non-human animals in the present invention include all vertebrates such as non-mammals (eg birds, amphibians, reptiles) and mammals such as non-human primates, livestock and/or domesticated animals (eg sheep, dogs) , cats, cows, pigs, etc.).
  • the present invention provides methods of preventing, alleviating and/or treating graft-versus-host disease following hematopoietic stem cell transplantation, comprising administering to an individual in need thereof an effective amount of a compound of formula (I) or a pharmacy thereof Acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs of:
  • Ring A is The above group is attached to the pyrimidine ring through one of the two positions marked with * or **, and the other position is attached to the carbonyl group;
  • R is selected from H and C 1-6 alkyl
  • R 2 is selected from H and C 1-6 alkyl
  • R 3 , R 4 , R 7 and R 8 at each occurrence are each independently selected from H, halogen, -NR 5 R 6 , -OH, C 1-6 alkyl and -OR 5 ;
  • alkylene, alkyl, alkenyl, cycloalkyl, heterocyclyl, aryl, heteroaryl and arylalkyl are each optionally substituted at each occurrence with one or more substituents independently selected from halogen, C 1 Substituent substitution of -6 alkyl and -OR 5;
  • R 5 and R 6 at each occurrence are each independently selected from H, C 1-6 alkyl, C 3-10 cyclohydrocarbyl, 3-10 membered heterocyclyl, C 6-10 aryl, 5-14 membered Heteroaryl and C 6-12 aralkyl;
  • n is independently at each occurrence an integer of 0, 1, 2 or 3;
  • n is independently an integer of 0, 1 or 2.
  • Ring A is The above groups are attached to the pyrimidine ring through the positions marked with *, and to the carbonyl group through the positions marked with **, wherein R 10 is selected from H and C 1-6 alkyl, preferably H or methyl.
  • Ring A is preferably The above groups are attached to the pyrimidine ring through the positions marked with *, and to the carbonyl group through the positions marked with **.
  • R is H
  • R 2 is H.
  • R 5 and R 6 are each independently selected from H, methyl and ethyl at each occurrence.
  • R 3, R 4, R 7 and R 8 at each occurrence is independently selected from H, F, Cl, Br, I, -NH 2, -OH, methyl, trifluoromethyl methyl, -CH 2 -Ph, methoxy, ethoxy and -CH 2 OCH 3.
  • R 3 is H.
  • R 4 is selected from H and halogen (e.g. F, Cl, Br or I), preferably H or F.
  • halogen e.g. F, Cl, Br or I
  • R 7 is selected from H and halogen (e.g. F, Cl, Br or I), preferably H or F.
  • halogen e.g. F, Cl, Br or I
  • R 8 is H.
  • each occurrence of R 9 is independently selected from H, C 1-6 alkyl, C 3-10 cyclohydrocarbyl, 3-10 membered heterocyclyl, C 6-10 aryl, 5-14 membered heteroaryl and C 6-12 aralkyl, preferably H.
  • each occurrence of R 10 is independently selected from H and C 1-6 alkyl, preferably H, methyl, ethyl, n-propyl or isopropyl, most preferably H or methyl.
  • the present invention provides a method of preventing, ameliorating and/or treating graft-versus-host disease following hematopoietic stem cell transplantation, comprising administering to an individual in need thereof an effective amount of a compound of formula (II) or the same Pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs:
  • the present invention provides a method of preventing, ameliorating and/or treating graft-versus-host disease following hematopoietic stem cell transplantation, comprising administering to an individual in need thereof an effective amount of a compound of formula (III) or its Pharmaceutically acceptable salts, esters, stereoisomers, polymorphs, solvates, N-oxides, isotopically labeled compounds, metabolites or prodrugs:
  • R 10 is H or methyl, preferably methyl.
  • the compound has the following structure:
  • the compounds are prepared according to the methods disclosed in WO 2019/001572 A1, which is incorporated herein by reference.
  • the compound of formula (I), formula (II), or formula (III), or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate is administered once or in two, three or four divided doses.
  • the compound of formula (I), formula (II), or formula (III), or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxides, isotopically labeled compounds, metabolites or prodrugs are administered consecutively for at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, At least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, at least 22 days, at least 23 days days, at least 24 days, at least 25 days, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 1 year, or at least 2 years.
  • the compound of formula (I), formula (II), or formula (III), or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate One or more (eg, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) courses of administration of N-oxides, isotopically-labeled compounds, metabolites, or prodrugs, wherein each course For at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, at least 14 days, at least 15 days, at least 16 days, at least 17 days, at least 18 days, at least 19 days, at least 20 days, at least 21 days, at least 22 days, at least 23 days, at least 24 days, at least 25 days, at least 30 days, at least 35 days , at least 40 days, at least 45 days, or at least 50 days; and 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 days, two
  • the compound of formula (I), formula (II), or formula (III), or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxides, isotopically-labeled compounds, metabolites, or prodrugs are administered by injection (eg, intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection, including infusion) or transdermally; or by oral, buccal , nasal, transmucosal, topical, in the form of ophthalmic formulations or by inhalation.
  • the compound of formula (I), formula (II), or formula (III), or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxides, isotopically labeled compounds, metabolites or prodrugs to be selected from tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes , lotions, ointments, aqueous suspensions, injectable solutions, elixirs and syrups.
  • the present invention provides a compound of Formula (I), Formula (II) or Formula (III) above, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate thereof , use of an N-oxide, isotopically labeled compound, metabolite or prodrug in the preparation of a medicament for preventing, alleviating and/or treating graft-versus-host disease after hematopoietic stem cell transplantation.
  • the present invention provides a compound of Formula (I), Formula (II) or Formula (III) above, or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate thereof , N-oxide, isotope-labeled compounds, metabolites or prodrugs for use in the prevention, mitigation and/or treatment of graft-versus-host disease following hematopoietic stem cell transplantation.
  • the hematopoietic stem cell transplantation is an allogeneic hematopoietic stem cell transplantation.
  • the graft-versus-host disease is acute graft-versus-host disease or chronic graft-versus-host disease.
  • a compound of formula (I), or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide, isotopically labeled compound, metabolite, thereof is Or the prodrug is administered before, during and/or after hematopoietic stem cell transplantation.
  • the present invention encompasses any combination of the above embodiments.
  • Compound 128 used in the examples has the following structure, which was prepared according to the method disclosed in WO 2019/001572 A1.
  • HTRF KinEASE-STK S2 kit HTRF KinEASE-STK S2 kit (62ST2PEC) for 50 kinase assay IC.
  • the kinase ROCK2 (01-119) used in the reaction was purchased from Carna Biosciences.
  • a 10000 nM compound solution was prepared with 1 ⁇ kinase buffer with a DMSO concentration of 2.5%, and the compounds were further diluted step by step with the above-mentioned DMSO-containing kinase buffer to obtain 9 different concentrations of the test compound solutions.
  • 2 ⁇ M of substrate ie, 2 ⁇ L of 5 ⁇ STK-S2 substrate working solution
  • 2 ⁇ L of 5 ⁇ ROCK2 kinase working solution containing 1.4 ng ROCK2 kinase
  • 2 ⁇ L of 5 ⁇ ATP working solution was added to all reaction wells and incubate at room temperature for 2 hours.
  • ER emission Ratio, emission ratio
  • inhibition rate (ER positive-ER test compound) /(ER positive-ER negative)*100%
  • mice Male BALB/c recipient mice were subjected to 8.5 Gy total body irradiation (TBI) (Gammacell 40, NORPION International Ltd., irradiation source: 137 Cs).
  • TBI total body irradiation
  • a suspension of bone marrow cells (BMC) and spleen cells (SC) of C57BL/6 (H-2b) donor mice was prepared, and the suspension was prepared at 0.5 mL/mouse (containing allogeneic C57BL/6 (H -2b) donor mice were 10 ⁇ 10 6 bone marrow cells and 6.25 ⁇ 10 6 spleen cell was allocated) after infusion into irradiated BALB / c recipient mice by tail vein.
  • mice were randomly divided into two groups: model group (G2) and compound 007 group (G3).
  • An irradiation group (G4) that is, only receiving 137 Cs radiation, without infusion of the donor mouse cell suspension
  • a syngeneic bone marrow transplantation group (G1) to this group of mice through tail vein infusion containing syngeneic BALB / c mice of 10 ⁇ 10 6 bone marrow cells in suspension.
  • Mice in the model group, the irradiated group and the syngeneic bone marrow transplantation group were orally administered with vehicle (0.2% Tween-80 aqueous solution) once a day for 23 days.
  • the mice in the compound 007 group were orally administered 30 mpk of compound 007 once a day for 23 days from the first day of model establishment. Animal survival was recorded daily.
  • C57BL/6 B6 mouse bone marrow cells
  • GM-CSF granulocyte macrophage colony stimulating factor
  • IL-4 interleukin 4
  • DC dendritic cells

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Abstract

预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的方法，其包括向需要其的个体给药有效量的式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药。

Description

治疗造血干细胞移植后的移植物抗宿主病的方法 发明领域
本发明属于生物医药领域，并具体涉及预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的方法，其包括向需要其的个体给药有效量的本申请中的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药。
发明背景
白血病等造血器官肿瘤的治疗首先是利用抗癌剂进行化疗，对通过通常的化疗难以治愈、或治愈可能性小的患者需要进行造血干细胞(例如外周血干细胞、骨髓细胞等)移植。但是，造血干细胞移植后会引发数种并发症，其中最主要的并发症为与造血干细胞移植有关的移植物抗宿主病(GVHD)。
为有助于GVHD诊断，美国国立卫生研究院(NIH)在2005年和2014年发布了指南和分类***，详细描述了GVHD的两个主要分类：急性移植物抗宿主病(aGVHD)和慢性移植物抗宿主病(cGVHD)。一般移植后100天以内发生的为aGVHD，100天以后发生的为cGVHD。
aGVHD发病率很高，多达50％的经历了造血干细胞移植的患者会发生aGVHD。aGVHD主要累及皮肤、胃肠道及肝，少数情况下也可累及其他脏器。通常仅发生皮肤损伤不会威胁生命；如果发生内脏损伤，则会出现重度黄疸、顽固性腹泻和血便、肠绞痛，以及严重的全身症状。在所有aGVHD患者中，高达60％的患者出现肝或胃肠道受损，死亡率达85％。
据估计，50％的经历了异基因造血干细胞移植的患者会发生cGVHD。cGVHD在移植后期发展，其可累及多种器官，如皮肤、嘴巴、眼睛、胃肠道、肝、肺、关节、筋膜以及生殖道等。
发明概述
在一个方面中，本发明提供预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的方法，其包括向需要其的个体给药有效量的式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药：
Figure PCTCN2021107575-appb-000001
其中：
环A为
Figure PCTCN2021107575-appb-000002
以上基团通过*或**标记的两个位置之一与嘧啶环连接，并且另一位置与羰基连接；
R选自H和C 1-6烷基；
R 1为
Figure PCTCN2021107575-appb-000003
R 2选自H和C 1-6烷基；
R 3、R 4、R 7和R 8在每次出现时各自独立地选自H、卤素、-NR 5R 6、-OH、C 1-6烷基和-OR 5；
R 9和R 10在每次出现时各自独立地选自H、卤素、C 1-6烷基、C 2-6烯基、C 3-10环烃基、3-10元杂 环基、C 6-10芳基、5-14元杂芳基、C 6-12芳烷基、-C(＝O)R 5和-C 1-6亚烷基-O(P＝O)(OH) 2；
上述亚烷基、烷基、烯基、环烃基、杂环基、芳基、杂芳基和芳烷基在每次出现时各自任选地被一个或多个独立地选自卤素、C 1-6烷基和-OR 5的取代基取代；
R 5和R 6在每次出现时各自独立地选自H、C 1-6烷基、C 3-10环烃基、3-10元杂环基、C 6-10芳基、5-14元杂芳基和C 6-12芳烷基；
m在每次出现时各自独立地为0、1、2或3的整数；并且
n在每次出现时各自独立地为0、1或2的整数。
在另一方面中，本发明提供上述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药在制备用于预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的药物中的用途。
在另一方面中，本发明提供上述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药，其用于预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病。
附图简要说明
图1显示实施例2中小鼠的生存率。
图2显示实施例3中测试的化合物007对炎症因子的抑制作用(与对照组相比，*P＜0.05，***P＜0.001，****P＜0.0001)。
发明详细描述
定义
除非在下文中另有定义，本文中所用的所有技术术语和科学术语的含义意图与本领域技术人员通常所理解的相同。提及本文中使用的技术意图指在本领域中通常所理解的技术，包括那些对本领域技术人员显而易见的技术的变化或等效技术的替换。虽然相信以下术语对于本领域技术人员很好理解，但仍然阐述以下定义以更好地解释本发明。
术语“包括”、“包含”、“具有”、“含有”或“涉及”及其在本文中的其它变体形式为包含性的(inclusive)或开放式的，且不排除其它未列举的元素或方法步骤。
如本文中所使用，术语“亚烷基”表示饱和二价烃基，优选表示具有1、2、3、4、5或6个碳原子的饱和二价烃基，例如亚甲基、亚乙基、亚丙基或亚丁基。
如本文中所使用，术语“烷基”定义为线性或支化饱和脂肪族烃。在一些实施方案中，烷基具有1至12个，例如1至6个碳原子。例如，如本文中所使用，术语“C 1-6烷基”指1至6个碳原子的线性或支化的基团(例如甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、异戊基、新戊基或正己基)，其任选地被1或多个(诸如1至3个)适合的取代基如卤素取代(此时该基团被称作“卤代烷基”)(例如CH 2F、CHF 2、CF 3、CCl 3、C 2F 5、C 2Cl 5、CH 2CF 3、CH 2Cl或-CH 2CH 2CF 3等)。术语“C 1-4烷基”指1至4个碳原子的线性或支化的脂肪族烃链(即甲基、乙基、正丙基、异丙基、正丁基、异丁基、仲丁基或叔丁基)。
如本文中所使用，术语“烯基”意指线性的或支化的单价烃基，其包含一个双键，且具有2-6个碳原子(“C 2-6烯基”)。所述烯基为例如乙烯基、1-丙烯基、2-丙烯基、2-丁烯基、3-丁烯基、2-戊烯基、3-戊烯基、4-戊烯基、2-己烯基、3-己烯基、4-己烯基、5-己烯基、2-甲基-2-丙烯基和4-甲基-3-戊烯基。当本发明的化合物含有亚烯基时，所述化合物可以纯E(异侧(entgegen))形式、纯Z(同侧(zusammen))形式或其任意混合物形式存在。
如本文中所使用，术语“炔基”表示包含一个或多个三键的单价烃基，其优选具有2、3、4、5或6个碳原子，例如乙炔基或丙炔基。
如本文中所使用，术语“环烷基”指饱和的单环或多环(诸如双环)烃环(例如单环，诸如环丙基、环丁基、环戊基、环己基、环庚基、环辛基、环壬基，或双环，包括螺环、稠合或桥连***(诸如双环[1.1.1]戊基、双环[2.2.1]庚基、双环[3.2.1]辛基或双环[5.2.0]壬基、十氢化萘基等))，其任选地被1或多个(诸如1至3个)适合的取代基取代。所述环烷基具有3至15个碳原子。例如，术语“C 3-6环烷基”指3至6个成环碳原子的饱和的单环或多环(诸如双环)烃环(例如环丙基、环丁基、环戊基或环己基)，其任选地被1或多个(诸如1至3个)适合的取代基取代，例如甲基取代的环丙基。
如本文中所使用，术语“亚环烃基”、“环烃基”和“烃环”是指具有例如3-10个(适合地具有3-8个， 更适合地具有3-6个)环碳原子的饱和(即，“亚环烷基”和“环烷基”)或不饱和的(即在环内具有一个或多个双键和/或三键)单环或多环烃环，其包括但不限于(亚)环丙基(环)、(亚)环丁基(环)、(亚)环戊基(环)、(亚)环己基(环)、(亚)环庚基(环)、(亚)环辛基(环)、(亚)环壬基(环)、(亚)环己烯基(环)等。
如本文中所使用，术语“杂环基”、“亚杂环基”和“杂环”是指具有例如3-10个(适合地具有3-8个，更适合地具有3-6个)环原子、其中至少一个环原子是选自N、O和S的杂原子且其余环原子是C的饱和(即，杂环烷基)或部分不饱和的(即在环内具有一个或多个双键和/或三键)环状基团。例如，“3-10元(亚)杂环(基)”是具有2-9个(如2、3、4、5、6、7、8或9个)环碳原子和独立地选自N、O和S的一个或多个(例如1个、2个、3个或4个)杂原子的饱和或部分不饱和(亚)杂环(基)。亚杂环基和杂环(基)的实例包括但不限于：(亚)环氧乙烷基、(亚)氮丙啶基、(亚)氮杂环丁基(azetidinyl)、(亚)氧杂环丁基(oxetanyl)、(亚)四氢呋喃基、(亚)二氧杂环戊烯基(dioxolinyl)、(亚)吡咯烷基、(亚)吡咯烷酮基、(亚)咪唑烷基、(亚)吡唑烷基、(亚)吡咯啉基、(亚)四氢吡喃基、(亚)哌啶基、(亚)吗啉基、(亚)二噻烷基(dithianyl)、(亚)硫吗啉基、(亚)哌嗪基或(亚)三噻烷基(trithianyl)。所述基团也涵盖双环***，包括螺环、稠合或桥连***(诸如8-氮杂螺[4.5]癸烷、3，9-二氮杂螺[5.5]十一烷、2-氮杂双环[2.2.2]辛烷等)。亚杂环基和杂环(基)可任选地被一个或多个(例如1个、2个、3个或4个)适合的取代基取代。
如本文中所使用，术语“(亚)芳基”和“芳环”指具有共轭π电子***的全碳单环或稠合环多环芳族基团。例如，如本文中所使用，术语“C 6-10(亚)芳基”和“C 6-10芳环”意指含有6至10个碳原子的芳族基团，诸如(亚)苯基(苯环)或(亚)萘基(萘环)。(亚)芳基和芳环任选地被1或多个(诸如1至3个)适合的取代基(例如卤素、-OH、-CN、-NO 2、C 1-6烷基等)取代。
如本文中所使用，术语“(亚)杂芳基”和“杂芳环”指单环、双环或三环芳族环系，其具有5、6、8、9、10、11、12、13或14个环原子，特别是1或2或3或4或5或6或9或10个碳原子，且其包含至少一个可以相同或不同的杂原子(所述杂原子是例如氧、氮或硫)，并且，另外在每一种情况下可为苯并稠合的。特别地，“(亚)杂芳基”或“杂芳环”选自(亚)噻吩基、(亚)呋喃基、(亚)吡咯基、(亚)噁唑基、(亚)噻唑基、(亚)咪唑基、(亚)吡唑基、(亚)异噁唑基、(亚)异噻唑基、(亚)噁二唑基、(亚)***基、(亚)噻二唑基等，以及它们的苯并衍生物；或(亚)吡啶基、(亚)哒嗪基、(亚)嘧啶基、(亚)吡嗪基、(亚)三嗪基等，以及它们的苯并衍生物。
如本文中所使用，术语“芳烷基”优选表示芳基或杂芳基取代的烷基，其中所述芳基、杂芳基和烷基如本文中所定义。通常，所述芳基可具有6-14个碳原子，所述杂芳基可具有5-14个环原子，并且所述烷基可具有1-6个碳原子。示例性芳烷基包括但不限于苄基、苯基乙基、苯基丙基、苯基丁基。
如本文中所使用，术语“卤代”或“卤素”基团定义为包括F、Cl、Br或I。
如本文中所使用，术语“含氮杂环”指饱和或不饱和的单环或双环基团，其在环中具有2、3、4、5、6、7、8、9、10、11、12或13个碳原子和至少一个氮原子，其还可任选地包含一个或多个(例如一个、两个、三个或四个)选自N、O、C＝O、S、S＝O和S(＝O) 2的环成员，其通过所述含氮杂环中的氮原子以及任一其余环原子与分子的其余部分连接，所述含氮杂环任选地为苯并稠合的，并且优选通过所述含氮杂环中的氮原子以及所稠合的苯环中的任一碳原子与分子的其余部分连接。
术语“取代”指所指定的原子上的一个或多个(例如一个、两个、三个或四个)氢被从所指出的基团的选择代替，条件是未超过所指定的原子在当前情况下的正常原子价并且所述取代形成稳定的化合物。取代基和/或变量的组合仅仅当这种组合形成稳定的化合物时才是允许的。
如果取代基被描述为“任选地被取代”，则取代基可(1)未被取代或(2)被取代。如果取代基的碳被描述为任选地被取代基列表中的一个或多个取代，则碳上的一个或多个氢(至存在的任何氢的程度)可单独和/或一起被独立地选择的任选的取代基替代。如果取代基的氮被描述为任选地被取代基列表中的一个或多个取代，则氮上的一个或多个氢(至存在的任何氢的程度)可各自被独立地选择的任选的取代基替代。
如果取代基被描述为“独立地选自”一组，则各取代基独立于另一者被选择。因此，各取代基可与另一(其他)取代基相同或不同。
如本文中所使用，术语“一个或多个”意指在合理条件下的1个或超过1个，例如2个、3个、4个、5个或10个。
除非指明，否则如本文中所使用，取代基的连接点可来自取代基的任意适宜位置。
当取代基的键显示为穿过环中连接两个原子的键时，则这样的取代基可键连至该可取代的环中的任一成环原子。
本发明还包括所有药学上可接受的同位素标记的化合物，其与本发明的化合物相同，除了一个或多个原子被具有相同原子序数但原子质量或质量数不同于在自然界中占优势的原子质量或质量数 的原子替代。适合包含入本发明的化合物中的同位素的实例包括(但不限于)氢的同位素(例如氘( 2H)、氚( 3H))；碳的同位素(例如 11C、 13C及 14C)；氯的同位素(例如 36Cl)；氟的同位素(例如 18F)；碘的同位素(例如 123I及 125I)；氮的同位素(例如 13N及 15N)；氧的同位素(例如 15O、 17O及 18O)；磷的同位素(例如 32P)；及硫的同位素(例如 35S)。某些同位素标记的本发明的化合物(例如掺入放射性同位素的那些)可用于药物和/或底物组织分布研究(例如分析)中。放射性同位素氚(即 3H)及碳-14(即 14C)因易于掺入且容易检测而特别可用于该目的。用正电子发射同位素(例如 11C、 18F、 15O及 13N)进行取代可在正电子发射断层显像术(PET)研究中用于检验底物受体占据情况。被同位素标记的本发明的化合物可通过与描述于随附路线和/或实施例及制备中的那些类似的方法通过使用适当的被同位素标记的试剂代替之前采用的非标记的试剂来制备。本发明的药学上可接受的溶剂合物包括其中结晶溶剂可被同位素取代的那些，例如，D 2O、丙酮-d 6或DMSO-d 6。
术语“立体异构体”表示由于至少一个不对称中心形成的异构体。在具有一个或多个(例如一个、两个、三个或四个)不对称中心的化合物中，其可产生外消旋混合物、单一对映异构体、非对映异构体混合物和单独的非对映异构体。特定个别分子也可以几何异构体(顺式/反式)存在。类似地，本发明的化合物可以两种或更多种处于快速平衡的结构不同的形式的混合物(通常称作互变异构体)存在。互变异构体的代表性实例包括酮-烯醇互变异构体、苯酚-酮互变异构体、亚硝基-肟互变异构体、亚胺-烯胺互变异构体等。要理解，本申请的范围涵盖所有这样的以任意比例(例如60％、65％、70％、75％、80％、85％、90％、95％、96％、97％、98％、99％)的异构体或其混合物。
本文中可使用实线
Figure PCTCN2021107575-appb-000004
实楔形
Figure PCTCN2021107575-appb-000005
或虚楔形
Figure PCTCN2021107575-appb-000006
描绘本发明的化合物的化学键。使用实线以描绘键连至不对称碳原子的键欲表明，包括该碳原子处的所有可能的立体异构体(例如，特定的对映异构体、外消旋混合物等)。使用实或虚楔形以描绘键连至不对称碳原子的键欲表明，存在所示的立体异构体。当存在于外消旋混合物中时，使用实及虚楔形以定义相对立体化学，而非绝对立体化学。除非另外指明，否则本发明的化合物意欲可以立体异构体(其包括顺式及反式异构体、光学异构体(例如R及S对映异构体)、非对映异构体、几何异构体、旋转异构体、构象异构体、阻转异构体及其混合物)的形式存在。本发明的化合物可表现一种以上类型的异构现象，且由其混合物(例如外消旋混合物及非对映异构体对)组成。
本发明涵盖本发明的化合物的所有可能的结晶形式或多晶型物，其可为单一多晶型物或多于一种多晶型物的任意比例的混合物。
还应当理解，本发明的某些化合物可以游离形式存在用于治疗，或适当时，以其药学上可接受的衍生物形式存在。在本发明中，药学上可接受的衍生物包括但不限于，药学上可接受的盐、酯、溶剂合物、N-氧化物、代谢物或前药，在将它们向需要其的患者给药后，能够直接或间接提供本发明的化合物或其代谢物或残余物。因此，当在本文中提及“本发明的化合物”时，也意在涵盖化合物的上述各种衍生物形式。
本发明的化合物的药学上可接受的盐包括其酸加成盐及碱加成盐。
适合的酸加成盐由形成药学可接受盐的酸来形成。实例包括乙酸盐、己二酸盐、天冬氨酸盐、苯甲酸盐、苯磺酸盐、碳酸氢盐/碳酸盐、硫酸氢盐/硫酸盐、硼酸盐、樟脑磺酸盐、柠檬酸盐、环己氨磺酸盐、乙二磺酸盐、乙磺酸盐、甲酸盐、延胡索酸盐、葡庚糖酸盐、葡糖酸盐、葡糖醛酸盐、六氟磷酸盐、海苯酸盐、盐酸盐/氯化物、氢溴酸盐/溴化物、氢碘酸盐/碘化物、羟乙基磺酸盐、乳酸盐、苹果酸盐、顺丁烯二酸盐、丙二酸盐、甲磺酸盐、甲基硫酸盐、萘甲酸盐(naphthylate)、2-萘磺酸盐、烟酸盐、硝酸盐、乳清酸盐、草酸盐、棕榈酸盐、双羟萘酸盐、磷酸盐/磷酸氢盐/磷酸二氢盐、焦谷氨酸盐、糖二酸盐、硬脂酸盐、丁二酸盐、单宁酸盐、酒石酸盐、甲苯磺酸盐、三氟乙酸盐及昔萘酸盐(xinofoate)。
适合的碱加成盐由形成药学可接受盐的碱来形成。实例包括铝盐、精氨酸盐、苄星青霉素盐、钙盐、胆碱盐、二乙胺盐、二乙醇胺盐、甘氨酸盐、赖氨酸盐、镁盐、葡甲胺盐、乙醇胺盐、钾盐、钠盐、氨丁三醇盐及锌盐。
适合的盐的综述参见Stahl及Wermuth的“Handbook of Pharmaceutical Salts：Properties，Selection，and Use”(Wiley-VCH，2002)。用于制备本发明的化合物的药学上可接受的盐的方法为本领域技术人员已知的。
如本文中所使用，术语“酯”意指衍生自本申请中各个通式化合物的酯，其包括生理上可水解的酯(可在生理条件下水解以释放游离酸或醇形式的本发明的化合物)。本发明的化合物本身也可以是酯。
本发明的化合物可以溶剂合物(优选水合物)的形式存在，其中本发明的化合物包含作为所述化合物晶格的结构要素的极性溶剂，特别是例如水、甲醇或乙醇。极性溶剂特别是水的量可以化学计量 比或非化学计量比存在。
本领域技术人员会理解，由于氮需要可用的孤对电子来氧化成氧化物，因此并非所有的含氮杂环都能够形成N-氧化物；本领域技术人员会识别能够形成N-氧化物的含氮杂环。本领域技术人员还会认识到叔胺能够形成N-氧化物。用于制备杂环和叔胺的N-氧化物的合成方法是本领域技术人员熟知的，包括用过氧酸如过氧乙酸和间氯过氧苯甲酸(MCPBA)、过氧化氢、烷基过氧化氢如叔丁基过氧化氢、过硼酸钠和双环氧乙烷(dioxirane)如二甲基双环氧乙烷来氧化杂环和叔胺。这些用于制备N-氧化物的方法已在文献中得到广泛描述和综述，参见例如：T.L.Gilchrist，Comprehensive Organic Synthesis，vol.7，pp 748-750；A.R.Katritzky和A.J.Boulton，Eds.，Academic Press；以及G.W.H.Cheeseman和E.S.G.Werstiuk，Advances in Heterocyclic Chemistry，vol.22，pp 390-392，A.R.Katritzky和A.J.Boulton，Eds.，Academic Press。
在本发明的范围内还包括本发明的化合物的代谢物，即在给药本发明的化合物时体内形成的物质。这样的产物可由例如被给药的化合物的氧化、还原、水解、酰胺化、脱酰胺化、酯化、酶解等产生。因此，本发明包括本发明的化合物的代谢物，包括通过使本发明的化合物与哺乳动物接触足以产生其代谢产物的时间的方法制得的化合物。
本发明在其范围内进一步包括本发明的化合物的前药，其为自身可具有较小药理学活性或无药理学活性的本发明的化合物的某些衍生物当被给药至身体中或其上时可通过例如水解裂解转化成具有期望活性的本发明的化合物。通常这样的前药会是所述化合物的官能团衍生物，其易于在体内转化成期望的治疗活性化合物。关于前药的使用的其他信息可参见“Pro-drugs as Novel Delivery Systems”，第14卷，ACS Symposium Series(T.Higuchi及V.Stella)。本发明的前药可例如通过用本领域技术人员已知作为“前-部分(pro-moiety)(例如“Design of Prodrugs”，H.Bundgaard(Elsevier，1985)中所述)”的某些部分替代本发明的化合物中存在的适当官能团来制备。
本发明还涵盖含有保护基的本发明的化合物。在制备本发明的化合物的任何过程中，保护在任何有关分子上的敏感基团或反应基团可能是必需的和/或期望的，由此形成本发明的化合物的化学保护的形式。这可以通过常规的保护基实现，例如，在T.W.Greene & P.G.M.Wuts，Protective Groups in Organic Synthesis，John Wiley & Sons，1991中所述的那些保护基，这些参考文献通过援引加入本文。使用本领域已知的方法，在适当的后续阶段可以移除保护基。
术语“约”是指在所述数值的±10％范围内，优选±5％范围内，更优选±2％范围内。
术语“有效量”是指在施用条件下足以达到所需治疗效果的量，其导致病理学症状、疾病进展、与之相关的生理状况改善或诱导对前述疾病进行的抵抗力。
除非另外说明，否则如本文中所使用，术语“治疗(treating)”意指逆转、减轻、抑制这样的术语所应用的病症或病况或者这样的病症或病况的一或多种症状的进展，或预防这样的病症或病况或者这样的病症或病况的一或多种症状。
如本文所使用的“个体”包括人或非人动物。示例性人个体包括患有疾病(例如本文所述的疾病)的人个体(称为患者)或正常个体。本发明中“非人动物”包括所有脊椎动物，例如非哺乳动物(例如鸟类、两栖动物、爬行动物)和哺乳动物，例如非人灵长类、家畜和/或驯化动物(例如绵羊、犬、猫、奶牛、猪等)。
具体实施方式
在一些实施方案中，本发明提供预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的方法，其包括向需要其的个体给药有效量的式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药：
Figure PCTCN2021107575-appb-000007
其中：
环A为
Figure PCTCN2021107575-appb-000008
以上基团通过*或**标记的两个位置之一与嘧啶环连接，并且另一位置与羰基连接；
R选自H和C 1-6烷基；
R 1为
Figure PCTCN2021107575-appb-000009
R 2选自H和C 1-6烷基；
R 3、R 4、R 7和R 8在每次出现时各自独立地选自H、卤素、-NR 5R 6、-OH、C 1-6烷基和-OR 5；
R 9和R 10在每次出现时各自独立地选自H、卤素、C 1-6烷基、C 2-6烯基、C 3-10环烃基、3-10元杂环基、C 6-10芳基、5-14元杂芳基、C 6-12芳烷基、-C(＝O)R 5和-C 1-6亚烷基-O(P＝O)(OH) 2；
上述亚烷基、烷基、烯基、环烃基、杂环基、芳基、杂芳基和芳烷基在每次出现时各自任选地被一个或多个独立地选自卤素、C 1-6烷基和-OR 5的取代基取代；
R 5和R 6在每次出现时各自独立地选自H、C 1-6烷基、C 3-10环烃基、3-10元杂环基、C 6-10芳基、5-14元杂芳基和C 6-12芳烷基；
m在每次出现时各自独立地为0、1、2或3的整数；并且
n在每次出现时各自独立地为0、1或2的整数。
在优选的实施方案中，环A为
Figure PCTCN2021107575-appb-000010
以上基团通过*标记的位置与嘧啶环连接，并且通过**标记的位置与羰基连接，其中R 10选自H和C 1-6烷基，优选为H或甲基。
在优选的实施方案中，环A优选为
Figure PCTCN2021107575-appb-000011
以上基团通过*标记的位置与嘧啶环连接，并且通过**标记的位置与羰基连接。
在优选的实施方案中，R为H。
在优选的实施方案中，R 2为H。
在优选的实施方案中，R 5和R 6在每次出现时各自独立地选自H、甲基和乙基。
在优选的实施方案中，R 3、R 4、R 7和R 8在每次出现时各自独立地选自H、F、Cl、Br、I、-NH 2、-OH、甲基、三氟甲基、-CH 2-Ph、甲氧基、乙氧基和-CH 2OCH 3。
在优选的实施方案中，R 3为H。
在优选的实施方案中，R 4选自H和卤素(例如F、Cl、Br或I)，优选为H或F。
在优选的实施方案中，R 7选自H和卤素(例如F、Cl、Br或I)，优选为H或F。
在优选的实施方案中，R 8为H。
在优选的实施方案中，R 9和R 10在每次出现时各自独立地选自H、F、Cl、Br、甲基、乙基、正丙基、异丙基、乙烯基、环丙基、环丁基、环戊基、氧杂环丁烷基、单氟甲基、二氟甲基、三氟甲基、乙酰基、-CH 2CHF 2、-CH 2OH、-CH 2OCH 3、-CH 2CH 2OCH 3、-CH 2-O(P＝O)(OH) 2、
Figure PCTCN2021107575-appb-000012
Figure PCTCN2021107575-appb-000013
在优选的实施方案中，R 9在每次出现时各自独立地选自H、C 1-6烷基、C 3-10环烃基、3-10元杂环基、C 6-10芳基、5-14元杂芳基和C 6-12芳烷基，优选为H。
在优选的实施方案中，R 10在每次出现时各自独立地选自H和C 1-6烷基，优选为H、甲基、乙基、 正丙基或异丙基，最优选为H或甲基。
在优选的实施方案中，本发明提供预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的方法，其包括向需要其的个体给药有效量的式(II)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药：
Figure PCTCN2021107575-appb-000014
其中各基团如上文所定义。
在优选的实施方案中，本发明提供预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的方法，其包括向需要其的个体给药有效量的式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药：
Figure PCTCN2021107575-appb-000015
其中R 10为H或甲基，优选为甲基。
在优选的实施方案中，所述化合物具有以下结构：
Figure PCTCN2021107575-appb-000016
Figure PCTCN2021107575-appb-000017
在一些实施方案中，所述化合物根据WO 2019/001572 A1(将其通过援引加入本文)中公开的方法制备。
在一些实施方案中，将所述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药以约0.005mg/日至约5000mg/日的量，例如约0.005、0.05、0.5、5、10、20、30、40、50、100、150、200、250、300、350、400、450、500、550、600、650、700、750、800、850、900、950、1000、1500、2000、2500、3000、3500、4000、4500或5000mg/日的量给药。
在一些实施方案中，将所述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药以每日约1ng/kg至约200mg/kg、约1μg/kg至约100mg/kg或者约1mg/kg至约50mg/kg体重的量给药，例如以每日约1μg/kg、约10μg/kg、约25μg/kg、约50μg/kg、约75μg/kg、约100μg/kg、约125μg/kg、约150μg/kg、约175μg/kg、约200μg/kg、约225μg/kg、约250μg/kg、约275μg/kg、约300μg/kg、约325μg/kg、约350μg/kg、约375μg/kg、约400μg/kg、约425μg/kg、约450μg/kg、约475μg/kg、约500μg/kg、约525μg/kg、约550μg/kg、约575μg/kg、约600μg/kg、约625μg/kg、约650μg/kg、约675μg/kg、约700μg/kg、约725μg/kg、约750μg/kg、约775μg/kg、约800μg/kg、约825μg/kg、约850μg/kg、约875μg/kg、约900μg/kg、约925μg/kg、约950μg/kg、约975μg/kg、约1mg/kg、约5mg/kg、约10mg/kg、约15mg/kg、约20mg/kg、约25mg/kg、约30mg/kg、约35mg/kg、约40mg/kg、约45mg/kg、约50mg/kg、约60mg/kg、约70mg/kg、约80mg/kg、约90mg/kg、约100mg/kg、约125mg/kg、约150mg/kg、约175mg/kg、约200mg/kg或约300mg/kg体重的量给药。
在一些实施方案中，将所述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药的每日剂量一次性给予或分两次、三次或四次给予。
在一些实施方案中，将所述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药连续给药至少3天、至少4天、至少5天、至少6天、至少7天、至少8天、至少9天、至少10天、至少11天、至少12天、至少13天、至少14天、至少15天、至少16天、至少17天、至少18天、至少19天、至少20天、至少21天、至少22天、至少23天、至少24天、至少25天、至少1个月、至少2个月、至少3个月、至少4个月、至少5个月、至少6个月、至少1年或至少2年。
在一些实施方案中，将所述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药给药一个或多个(例如1、2、3、4、5、6、7、8、9或10个)疗程，其中每个疗程持续至少3天、至少4天、至少5天、至少6天、至少7天、至少8天、至少9天、至少10天、至少11天、至少12天、至少13天、至少14天、至少15天、至少16天、至少17天、至少18天、至少19天、至少20天、至少21天、至少22天、至少23天、至少24天、至少25天、至少30天、至少35天、至少40天、至少45天或至少50天；并且每两个疗程之间间隔0、1、2、3、4、5、6、7、8、9、10天、两周、三周或四周。
在一些实施方案中，将所述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药通过注射(如静脉内、动脉内、皮下、腹膜内、肌内注射，包括滴注)或经皮给药；或通过口服、含服、经鼻、透粘膜、局部、以眼用制剂的形式或通过吸入给药。
在一些实施方案中，将所述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药以选自片剂、胶囊剂、锭剂、硬糖剂、散剂、喷雾剂、乳膏剂、软膏剂、栓剂、凝胶剂、糊剂、洗剂、软膏剂、水性混悬剂、可注射溶液剂、酏剂和糖浆剂的剂型给药。
在一些实施方案中，本发明提供上述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药在制备用于预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的药物中的用途。
在一些实施方案中，本发明提供上述式(I)、式(II)或式(III)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药，其用于预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病。
在一些实施方案中，所述造血干细胞移植是同种异体造血干细胞移植。
在一些实施方案中，所述移植物抗宿主病为急性移植物抗宿主病或慢性移植物抗宿主病。
在一些实施方案中，将式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药在造血干细胞移植之前、期间和/或之后给药。
本发明涵盖以上实施方案的任意组合。
实施例
为了使本发明的目的和技术方案更加清楚，以下结合具体实施例进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。并且，下列实施例中未提及的具体实验方法，均按照常规实验方法进行。
实施例中所使用化合物128具有以下结构，其根据WO 2019/001572 A1中公开的方法制备。
Figure PCTCN2021107575-appb-000018
实施例1.ROCK2激酶活性检测
采用商品化的CISBIO激酶检测试剂盒HTRF KinEASE-STK S2试剂盒(62ST2PEC)进行激酶IC 50测定。反应所用的激酶ROCK2(01-119)购自Carna Biosciences。
实验开始前，根据需要，按照激酶检测试剂盒说明书用相应试剂配制如下工作液：1×激酶缓冲液，5×STK-S2底物工作液(1.5μM)和5×ATP工作液(1.5μM)、5×ROCK2激酶工作液、4×抗生蛋白链菌素-XL665工作液、4×STK-Ab-穴状化合物2检测液，然后进行如下操作。
采用DMSO浓度为2.5％的1×激酶缓冲液配制得到10000nM的化合物溶液，采用上述含DMSO的激酶缓冲液再对化合物进行逐级稀释，得到9个不同浓度的待测化合物溶液。除测试化合物孔外，设置阳性孔(含有除化合物之外的所有试剂)和阴性孔(含有除待测化合物和激酶外的所有试剂)。除对照孔(阳性孔和阴性孔)外，向所有反应孔中加入4μL的待测化合物溶液，向对照孔中加入4μL 2.5％的DMSO溶液。然后，再向所有反应孔中加入2μM底物(即2μL 5×STK-S2底物工作液)。向除阴性孔外的所有反应孔中加入2μL的5×ROCK2激酶工作液(含1.4ng ROCK2激酶)，阴性孔用2μL的1×激酶缓冲液补足体积。向所有反应孔中加入2μL 5×ATP工作液，室温孵育2小时。待激酶反应结束后，向所有反应孔中加入5μL 4×抗生蛋白链菌素-XL665工作液，混匀后立即加入5μL 4×STK-Ab-穴状化合物2检测液，室温孵育1小时后，用ENVISION(Perkinelmer)仪器检测荧光信号(激发波长为320nm，发射波长为665nm和615nm)。通过各孔荧光强度值计算出每个孔中的抑制率：ER(Emission Ratio，发射光比率)＝(665nm处荧光强度/615nm处荧光强度)；抑制率＝(ER阳性-ER待测化合物)/(ER阳性-ER阴性)*100％，用软件PRISM 5.0绘制曲线图，同时用PRISM 5.0拟合得到各待 测化合物的半数抑制浓度(IC 50)。化合物IC 50值如下表所示。
表1
化合物 ROCK2 IC 50nM 化合物 ROCK2 IC 50nM
化合物006 34 化合物011 9
化合物007 33 化合物020 44
化合物008 24 化合物021 45
化合物009 12 化合物022 75
化合物010 61 化合物128 27
实施例2.在小鼠中的治疗作用
使雄性BALB/c受体小鼠接受8.5Gy全身性辐照(TBI)(Gammacell 40，NORPION国际有限公司，辐照源： 137Cs)。制备C57BL/6(H-2b)供体小鼠骨髓细胞(BMC)和脾脏细胞(SC)的悬浮液，将所述悬浮液以0.5mL/只(其中含同种异系C57BL/6(H-2b)供体小鼠的10×10 6个骨髓细胞和6.25×10 6个脾脏细胞)通过尾静脉输注至经过辐照的BALB/c受体小鼠。随后将BALB/c小鼠随机分为2组：模型组(G2)、化合物007组(G3)。另设辐照组(G4)(即仅接受 137Cs射线辐照，未输注供体小鼠细胞悬浮液)和同系骨髓移植组(G1)(向该组小鼠通过尾静脉输注含有同系BALB/c小鼠的10×10 6个骨髓细胞的悬浮液)。G1-G4组，每组9只小鼠。将模型组、辐照组和同系骨髓移植组的小鼠经口灌胃给予溶媒(0.2％吐温-80水溶液)，每天一次，持续23天。将化合物007组的小鼠从模型建立第一天开始经口灌胃给与30mpk化合物007，每天1次，持续23天。每天记录动物的生存率。
生存率结果如图1所示。结果表明，与模型组相比，化合物007组的小鼠具有显著提高小鼠生存率。
实施例3.同种异体混合淋巴细胞反应
分离C57BL/6(B6)小鼠骨髓细胞，加入粒细胞巨噬细胞集落刺激因子(GM-CSF，20ng/mL，Biolegend，713704)和白介素4(IL-4，10ng/mL，Biolegend，10715004)培养7天，随后加入20ng/mL的脂多糖(LPS)培养24小时，收集树突状细胞(DC)。用商品化试剂盒(达科为，480039)提取分离BALB/c小鼠CD4
Figure PCTCN2021107575-appb-000019
T细胞。将10 5个CD4
Figure PCTCN2021107575-appb-000020
T细胞和25×10 3DC细胞进行混合培养。向混合细胞内分别加入4个浓度(125nM、250nM、500nM和1000nM)的化合物007，另设立对照组(加入同等剂量的培养基)。培养5天后收集上清液，使用试剂盒进行炎症因子检测(IL-6(达优，1210602)、TNF-α(达优，1217202)、IFN-γ(达优，1210002)、IL-2(达优，1210202))。
实验结果如图2所示。结果表明，化合物007显著降低了炎症因子的水平。
除本文中描述的那些外，根据前述描述，本发明的各种修改对本领域技术人员而言会是显而易见的。这样的修改也意图落入所附权利要求书的范围内。本申请中所引用的各参考文献(包括所有专利、专利申请、期刊文章、书籍及任何其它公开)均以其整体援引加入本文。

Claims (13)

  1. 预防、缓解和/或治疗造血干细胞移植后的移植物抗宿主病的方法，其包括向需要其的个体给药有效量的式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药：
    Figure PCTCN2021107575-appb-100001
    其中：
    环A为
    Figure PCTCN2021107575-appb-100002
    以上基团通过*或**标记的两个位置之一与嘧啶环连接，并且另一位置与羰基连接；优选地，环A为
    Figure PCTCN2021107575-appb-100003
    以上基团通过*标记的位置与嘧啶环连接，并且通过**标记的位置与羰基连接，其中R 10选自H和C 1-6烷基，优选为H或甲基；
    R选自H和C 1-6烷基；
    R 1为
    Figure PCTCN2021107575-appb-100004
    R 2选自H和C 1-6烷基；
    R 3、R 4、R 7和R 8在每次出现时各自独立地选自H、卤素(例如F、Cl、Br或I)、-NR 5R 6、-OH、C 1-6烷基和-OR 5；
    R 9和R 10在每次出现时各自独立地选自H、卤素、C 1-6烷基(例如甲基)、C 2-6烯基、C 3-10环烃基、3-10元杂环基、C 6-10芳基、5-14元杂芳基、C 6-12芳烷基、-C(＝O)R 5和-C 1-6亚烷基-O(P＝O)(OH) 2；
    上述亚烷基、烷基、烯基、环烃基、杂环基、芳基、杂芳基和芳烷基在每次出现时各自任选地被一个或多个独立地选自卤素、C 1-6烷基和-OR 5的取代基取代；
    R 5和R 6在每次出现时各自独立地选自H、C 1-6烷基、C 3-10环烃基、3-10元杂环基、C 6-10芳基、5-14元杂芳基和C 6-12芳烷基；
    m在每次出现时各自独立地为0、1、2或3的整数；并且
    n在每次出现时各自独立地为0、1或2的整数。
  2. 权利要求1的方法，其中所述化合物具有式(II)的结构：
    Figure PCTCN2021107575-appb-100005
    其中各基团如权利要求1中所定义；
    优选地，所述化合物具有式(III)的结构：
    Figure PCTCN2021107575-appb-100006
    其中R 10为H或甲基，优选为甲基。
  3. 权利要求1或2的方法，其中所述化合物具有以下结构：
    Figure PCTCN2021107575-appb-100007
  4. 权利要求1-3中任一项的方法，其中将所述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药以约0.005mg/日至 约5000mg/日的量，例如约0.005、0.05、0.5、5、10、20、30、40、50、100、150、200、250、300、350、400、450、500、550、600、650、700、750、800、850、900、950、1000、1500、2000、2500、3000、3500、4000、4500或5000mg/日的量给药。
  5. 权利要求1-3中任一项的方法，其中将所述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药以每日约1ng/kg至约200mg/kg、约1μg/kg至约100mg/kg或者约1mg/kg至约50mg/kg体重的量给药，例如以每日约1μg/kg、约10μg/kg、约25μg/kg、约50μg/kg、约75μg/kg、约100μg/kg、约125μg/kg、约150μg/kg、约175μg/kg、约200μg/kg、约225μg/kg、约250μg/kg、约275μg/kg、约300μg/kg、约325μg/kg、约350μg/kg、约375μg/kg、约400μg/kg、约425μg/kg、约450μg/kg、约475μg/kg、约500μg/kg、约525μg/kg、约550μg/kg、约575μg/kg、约600μg/kg、约625μg/kg、约650μg/kg、约675μg/kg、约700μg/kg、约725μg/kg、约750μg/kg、约775μg/kg、约800μg/kg、约825μg/kg、约850μg/kg、约875μg/kg、约900μg/kg、约925μg/kg、约950μg/kg、约975μg/kg、约1mg/kg、约5mg/kg、约10mg/kg、约15mg/kg、约20mg/kg、约25mg/kg、约30mg/kg、约35mg/kg、约40mg/kg、约45mg/kg、约50mg/kg、约60mg/kg、约70mg/kg、约80mg/kg、约90mg/kg、约100mg/kg、约125mg/kg、约150mg/kg、约175mg/kg、约200mg/kg或约300mg/kg体重的量给药。
  6. 权利要求1-5中任一项的方法，其中将所述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药的每日剂量一次性给予或分两次、三次或四次给予。
  7. 权利要求1-6中任一项的方法，其中将所述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药连续给药至少3天、至少4天、至少5天、至少6天、至少7天、至少8天、至少9天、至少10天、至少11天、至少12天、至少13天、至少14天、至少15天、至少16天、至少17天、至少18天、至少19天、至少20天、至少21天、至少22天、至少23天、至少24天、至少25天、至少1个月、至少2个月、至少3个月、至少4个月、至少5个月、至少6个月、至少1年或至少2年。
  8. 权利要求1-7中任一项的方法，其中将所述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药给药一个或多个(例如1、2、3、4、5、6、7、8、9或10个)疗程，其中每个疗程持续至少3天、至少4天、至少5天、至少6天、至少7天、至少8天、至少9天、至少10天、至少11天、至少12天、至少13天、至少14天、至少15天、至少16天、至少17天、至少18天、至少19天、至少20天、至少21天、至少22天、至少23天、至少24天、至少25天、至少30天、至少35天、至少40天、至少45天或至少50天；并且每两个疗程之间间隔0、1、2、3、4、5、6、7、8、9、10天、两周、三周或四周。
  9. 权利要求1-8中任一项的方法，其中将所述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药通过注射(如静脉内、动脉内、皮下、腹膜内、肌内注射，包括滴注)或经皮给药；或通过口服、含服、经鼻、透粘膜、局部、以眼用制剂的形式或通过吸入给药。
  10. 权利要求1-9中任一项的方法，其中将所述式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药以选自片剂、胶囊剂、锭剂、硬糖剂、散剂、喷雾剂、乳膏剂、软膏剂、栓剂、凝胶剂、糊剂、洗剂、软膏剂、水 性混悬剂、可注射溶液剂、酏剂和糖浆剂的剂型给药。
  11. 权利要求1-10中任一项的方法，其中所述造血干细胞移植是同种异体造血干细胞移植。
  12. 权利要求1-11中任一项的方法，其中所述移植物抗宿主病为急性移植物抗宿主病或慢性移植物抗宿主病。
  13. 权利要求1-12中任一项的方法，其中将式(I)的化合物或其药学上可接受的盐、酯、立体异构体、多晶型物、溶剂合物、N-氧化物、同位素标记的化合物、代谢物或前药在造血干细胞移植之前、期间和/或之后给药。
PCT/CN2021/107575 2020-07-22 2021-07-21 治疗造血干细胞移植后的移植物抗宿主病的方法 WO2022017412A1 (zh)

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CA3186564A CA3186564A1 (en) 2020-07-22 2021-07-21 Method for treating graft versus host disease caused by hematopoietic stem cell transplantation
BR112023001049A BR112023001049A2 (pt) 2020-07-22 2021-07-21 Método para tratamento de doença de enxerto versus hospedeiro causada por transplante de célula-tronco hematopoiética
JP2023504356A JP2023535035A (ja) 2020-07-22 2021-07-21 造血幹細胞移植によって引き起こされる移植片対宿主病を治療する方法
CN202180059344.8A CN116322670A (zh) 2020-07-22 2021-07-21 治疗造血干细胞移植后的移植物抗宿主病的方法
IL299985A IL299985A (en) 2020-07-22 2021-07-21 A method for treating graft-versus-Maschen disease caused by hematopoietic stem cell transplantation
EP21845732.3A EP4186508A1 (en) 2020-07-22 2021-07-21 Method for treating graft versus host disease caused by hematopoietic stem cell transplantation
US18/017,198 US20230285393A1 (en) 2020-07-22 2021-07-21 Method for treating graft versus host disease caused by hematopoietic stem cell transplantation
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