CN113135909A

CN113135909A - DPD inhibitor and preparation method, pharmaceutical composition and application thereof

Info

Publication number: CN113135909A
Application number: CN202010051408.7A
Authority: CN
Inventors: 曲显俊; 万升标; 崔淑香; 张国军
Original assignee: Beijing Shenlantai Pharmaceutical Technology Co ltd
Current assignee: Beijing Shenlantai Pharmaceutical Technology Co ltd
Priority date: 2020-01-17
Filing date: 2020-01-17
Publication date: 2021-07-20
Anticipated expiration: 2040-01-17
Also published as: CN113135909B

Abstract

The application relates to a novel DPD expression inhibitor, a preparation method thereof, a pharmaceutical composition and application thereof, and new application of a S1PR2 inhibitor compound, in particular to application of an S1PR2 inhibitor in inhibiting expression of dihydropyrimidine dehydrogenase DPD, especially application in reversing drug resistance appearing in tumor treatment by fluorouracil drugs.

Description

DPD inhibitor and preparation method, pharmaceutical composition and application thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to a novel DPD inhibitor, a preparation method, a pharmaceutical composition and application thereof, and new application of a sphingosine-1-type phosphate receptor (S1PR2) inhibitor. In particular to the application of an S1PR2 inhibitor in inhibiting the expression of dihydropyrimidine dehydrogenase DPD.

Background

Chemotherapy plays an extremely important role in cancer treatment. Since the synthesis of 5-fluorouracil by 1957Duschinsky et al, 5-fluorouracil and its derivatives have been the basis for chemotherapy of tumors of the digestive tract, head and neck and other tumors, such as lung and breast cancer. However, like other anticancer drugs, the development of tumor resistance to them is a major cause of the failure of cancer chemotherapy as described above. Indeed, dihydropyrimidine dehydrogenase (DPD) in tumor cells is the major rate-limiting enzyme for the degradation of 5-fluorouracil into cancer cells, which leads to drug resistance in tumor cells due to the degradation of intracellular 5-fluorouracil into inactive products. However, the mechanism for regulating DPD expression is still unknown, so that no mention is made about new target discovery and targeted design of 5-fluorouracil resistance reversal agents, which is an urgent need drug for clinical doctors to look ahead. Therefore, the key molecules for regulating DPD expression in the process of generating 5-fluorouracil resistance are confirmed and are important for designing a target of a drug resistance reversal agent.

Sphingosine-1-type phosphate receptors (S1PR2, also known as EDG-5, H218, AGR16, lpB2) are, as one of the members of the G protein-coupled receptor family, typical members of the rhodopsin (rhodopsin-like receptor, class a) family. The main functions reported in the current research are inhibiting endothelial cell migration, promoting insulin secretion, inhibiting tumor cell migration and the like. The clinically applied FTY720 can be combined with other targets S1PR1, S1PR3, S1PR4 and S1PR5 on the surface of inflammatory cell membranes in a phosphorylation mode through simulating an S1P structure so as to generate corresponding functions of inhibiting inflammatory factors, but the FTY720 is not combined with S1PR2 alone or is extremely poor in combination and cannot cause downstream effective signal transmission.

The S1PR2 inhibitors which are available at present mainly include pyridopyrazole compounds, diaryloxyphenyl compounds, pyridazine compounds, such as WO08154470, WO2011/041287, WO2011/159864, WO2013/148460, WO2014/158302, WO2016/191872, US2019/0127372, WO2013/047701, WO2017/148787, CN108699037, org.biomol.chem.13(2015)7928, bioorg.med.chem.lett.25(2015) 1479-1482, bioorg.med.chem.lett.25(2015) 4387-1209-4392, bioorg.med.chem.lett.26 (med) 1209-496, and the like, which are not described in terms of DPD cell expression. The above patent and non-patent documents are incorporated by reference herein in their entirety.

Disclosure of Invention

In a first aspect, the present application provides a compound of formula I, or a pharmaceutically acceptable salt, or tautomer thereof:

wherein:

R₁selected from optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocycloalkenyl;

R₂selected from hydrogen or optionally substituted alkyl;

each R₃Independently selected from hydrogen, deuterium, halogen, -CN, -NO₂Nitroso, carboxyl, sulfonic acid, optionally substituted sulfonyl, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl;

R₄、R₅and R₆Each independently selected from hydrogen, or optionally substituted alkyl;

ring B is an aromatic or heteroaromatic ring;

each R₇Independently selected from hydrogen, halogen, optionally substituted alkyl, optionally substituted alkoxy, or optionally substituted amino;

x is an integer from 1 to 2; and

y is an integer of 1,2 or 3.

In a second aspect, the present application provides the following compounds, or a pharmaceutically acceptable salt, or tautomer thereof:

in a third aspect, the present application provides a process for the preparation of a compound of the first aspect, comprising: step 1:

step 2:

and the following step 3-1 or step 3-2:

step 3-1:

step 3-2:

wherein R is₀And R₀' are each independently halogen; ring B, R₁、R₂、R₃、R₄、R₅、R₆、R₇X, and y are as described in the above first aspect.

In a fourth aspect, the present application provides a pharmaceutical composition comprising a compound of the first aspect described above, a pharmaceutically acceptable salt thereof, or a tautomer thereof.

In a fifth aspect, the present application provides the use of a compound of the first aspect above, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of the fourth aspect above, or a combination thereof with one or more other therapeutic agents, in the manufacture of a medicament for inhibiting the expression of DPD; or in the manufacture of a medicament for the prevention and/or treatment of a disease, disorder or condition that benefits from inhibition of DPD expression; or in the preparation of medicaments for preventing and/or treating tumors; or in the preparation of drugs for resisting and/or reversing drug resistance of fluorouracil drugs.

In a sixth aspect, the present application provides the use of an inhibitor of S1PR2, a pharmaceutically acceptable salt, or tautomer thereof, or pharmaceutical composition thereof, or combination thereof with one or more other therapeutic agents, in the manufacture of a medicament for inhibiting the expression of DPD; or in the manufacture of a medicament for the prevention and/or treatment of a disease, disorder or condition that benefits from inhibition of DPD expression; or in the preparation of a medicament for preventing and/or treating tumors, wherein the tumors have drug resistance to fluorouracil drugs; or in the preparation of drugs for resisting and/or reversing drug resistance of fluorouracil drugs.

In a seventh aspect, the present application provides a method for inhibiting DPD expression in a cell, comprising administering to said cell an effective amount of a compound of the first aspect, a pharmaceutically acceptable salt, or tautomer thereof, as described above, or a pharmaceutical composition of the fourth aspect, or combinations thereof with one or more other therapeutic agents, or an inhibitor of S1PR2, as described above, wherein said method is performed in vitro.

In an eighth aspect, the present application provides a method for combating/reversing drug resistance in a cell, comprising administering to said cell an effective amount of a compound according to the first aspect above, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition according to the fourth aspect above, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 according to the sixth aspect above, wherein said method is performed in vitro.

In a ninth aspect, the present application provides a method of screening for an anti-drug resistant oncology therapeutic drug, wherein the anti-drug resistant oncology therapeutic drug is an oncology therapeutic drug capable of resisting and/or reversing drug resistance to fluorouracil drugs, the method comprising:

providing a tumor cell line or cell culture expressing S1PR 2;

contacting a candidate with the tumor cell line or cell culture;

detecting the expression level of S1PR2 in the tumor cell line or cell culture after contact with the candidate and comparing to the expression level of S1PR2 in a control tumor cell line or cell culture not contacted with the candidate;

when the expression level of S1PR2 is lower than that of a control tumor cell line or cell culture, the candidate object can be used as an anti-drug-resistant tumor treatment drug.

In a tenth aspect, the present application provides a method of screening for an anti-drug resistant oncology therapeutic, wherein the anti-drug resistant oncology therapeutic is an oncology therapeutic capable of resisting and/or reversing drug resistance to fluorouracil drugs, comprising:

providing a DPD-expressing tumor cell line or cancer cell culture;

contacting a candidate with the tumor cell line or cancer cell culture;

detecting the expression level of DPD in said tumor cell line or cell culture following contact with the candidate and comparing the expression level of DPD with the expression level of DPD in a control tumor cell line or cell culture not contacted with the candidate;

when the DPD expression level is lower than a control tumor cell line or cell culture, indicating that the candidate is capable of acting as an anti-drug resistant tumor therapy;

wherein the candidate is selected from S1PR2 inhibitors.

In an eleventh aspect, the present application provides a kit comprising:

a compound of the first aspect above, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition of the fourth aspect above, or an inhibitor of S1PR2 of the sixth aspect above; and

optionally an antineoplastic agent.

In a twelfth aspect, the present application provides a compound of the first aspect, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition of the fourth aspect, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 of the sixth aspect, as described above, for use in inhibiting the expression of DPD.

In a thirteenth aspect, the present application provides a compound of the first aspect described above, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition of the fourth aspect described above, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 of the sixth aspect described above, for use in the prevention and/or treatment of a disease, disorder or condition that benefits from the inhibition of DPD expression.

In a fourteenth aspect, the present application provides a compound of the first aspect, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition of the fourth aspect, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 of the sixth aspect, for use in combatting and/or reversing drug resistance to fluorouracil drugs.

In a fifteenth aspect, the present application provides a method of inhibiting the expression of DPD, comprising administering to a subject (e.g., a mammal such as a human being) in need thereof a therapeutically effective amount of a compound of the first aspect, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition of the fourth aspect, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2, as described in the sixth aspect, above.

In a sixteenth aspect, the present application provides a method for the prevention and/or treatment of a disease, disorder or condition that benefits from inhibition of DPD expression, comprising administering to a subject (e.g. a mammal such as a human being) in need thereof a therapeutically effective amount of a compound of the first aspect described above, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of the fourth aspect described above, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 of the sixth aspect described above.

In a seventeenth aspect, the present application provides a method of combating and/or reversing drug resistance to fluorouracil-like drugs, comprising:

a fluorouracil drug is administered in combination with a compound according to the above first aspect of the present invention, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition according to the above fourth aspect, or an inhibitor of S1PR2 according to the above sixth aspect.

In an eighteenth aspect, the present application provides the use of a compound of the first aspect above, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of the fourth aspect above, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 of the sixth aspect above, for inhibiting the expression of DPD.

In a nineteenth aspect, the present application provides a compound of the first aspect, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition of the fourth aspect, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 of the sixth aspect, for use in the prevention and/or treatment of a disease, disorder, or condition that benefits from inhibition of DPD expression.

In a twentieth aspect, the present application provides a compound of the first aspect above, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition of the fourth aspect above, or a combination thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 of the sixth aspect above, for use in combating and/or reversing the resistance of fluorouracil drugs.

Drawings

Figure 1 shows the experimental results of in vitro inhibition of DPD expression for both the existing S1PR2 inhibitor and the newly synthesized compounds of the present disclosure.

FIG. 2 shows the tumor volume changes and the tumor size differences reversing 5-FU resistance in groups of nude mice during treatment with the S1PR2 inhibitor JTE-013 and the compounds 16-6.

FIG. 3 shows DPD expression differences in tumor, normal colon and liver tissues of each group of nude mice after material selection.

FIG. 4 shows the inhibition of DPD expression in vitro following knockdown of S1PR 2.

Detailed Description

Definition of

The following definitions and methods are provided to better define the present application and to guide those of ordinary skill in the art in the practice of the present application. Unless otherwise indicated, terms are to be understood in accordance with their ordinary usage by those of ordinary skill in the relevant art. All patent documents, academic papers, and other publications cited herein are incorporated by reference in their entirety.

The word "comprise" and its derivatives, such as "comprises" or "comprising," are to be construed in an open, non-exclusive sense, i.e., to mean "including, but not limited to.

The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

Numerical ranges indicating the number of carbon atoms are used herein to refer to each integer in the given range, e.g., "C₁-C₁₈"or" C_1-18By "is meant that the group may have 1,2,3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 carbon atoms.

The term "member" is intended to mean the number of skeleton atoms constituting a ring. For example, "3-12 membered" means that the number of backbone atoms constituting the ring is 3,4, 5, 6, 7, 8, 9, 10, 11 or 12.

The terms "halo", "halo" or "halogen" by themselves or as part of another substituent represent a fluorine (F), chlorine (Cl), bromine (Br) or iodine (I) atom.

The term "alkyl" refers to a straight or branched chain saturated hydrocarbon group, which may be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). Examples of alkyl groups include C_1-4Alkyl radical, C_1-6Alkyl radical, C_1-8Alkyl radical, C_1-10Alkyl radical, C_1-12Alkyl groups and the like, for example, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, s-butyl, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl) and the like; for example, the term "C_1-6Alkyl "refers to an alkyl group containing 1 to 6 (e.g., 1,2,3,4, 5, 6) carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl)Alkyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.).

The term "hydroxyalkyl" refers to-alkyl-OH, wherein "alkyl" is as defined above and "optionally substituted hydroxyalkyl" refers to an alkyl group in the group that is substituted or unsubstituted.

The term "alkoxy" refers to-O-alkyl, wherein "alkyl" is as defined above, and examples of "alkoxy" include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, isopentoxy. "optionally substituted alkoxy" means that the alkyl group in the group is substituted or unsubstituted.

The term "alkenyl" refers to a straight or branched chain unsaturated aliphatic hydrocarbon group having at least one double bond consisting of carbon and hydrogen atoms, such as C_2-6Alkenyl radical, C_2-4Alkenyl groups, and the like. Non-limiting examples of alkenyl groups include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1, 3-butadienyl, and the like. The term includes cis and trans isomers or mixtures of such isomers.

The term "alkynyl" refers to a straight or branched chain unsaturated aliphatic hydrocarbon group having at least one triple bond consisting of carbon and hydrogen atoms, such as C_2-6Alkynyl, C_2-4Alkynyl, C_2-3Alkynyl and the like. Non-limiting examples of alkynyl groups include, but are not limited to, ethynyl (-C ≡ CH), 1-propynyl (-C ≡ C-CH)₃) 2-propynyl (-CH)₂-C.ident.CH), 1, 3-butadiynyl (-C.ident.C-C.ident.CH), and the like.

The term "cycloalkyl" refers to a fully saturated nonaromatic ring of carbon and hydrogen atoms, preferably containing 1 or 2 rings. The cycloalkyl group may be a monocyclic, fused polycyclic, bridged or spiro ring structure. Non-limiting examples of cycloalkyl include, but are not limited to, C_3-18Cycloalkyl radical, C_3-16Cycloalkyl radical, C_3-12Cycloalkyl radical, C_3-10Cycloalkyl radical, C_3-8Cycloalkyl radical, C_3-7Cycloalkyl radical, C_3-6Cycloalkyl radicals and the like, e.g. cyclopropyl, cyclobutyl, cycloPentyl, cyclohexyl, cycloheptyl, spiro [3.3 ]]Heptyl, norbornyl (bicyclo [ 2.2.1)]Heptyl), bicyclo [2.2.2]Octyl, adamantyl, bicyclo [1.1.1]Pent-1-yl, and the like.

The term "heterocycloalkyl" refers to a cyclic group that is fully saturated and may exist as a single ring, a bridged ring, or a spiro ring. Unless otherwise indicated, the heterocyclic ring is typically a ring containing 1 to 5 (e.g., 1,2,3,4, 5) heteroatoms independently selected from sulfur, oxygen, and/or nitrogen. Examples of 3-membered heterocycloalkyl include, but are not limited to, oxiranyl, thietanyl, cycloazenyl, non-limiting examples of 4-membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thiabutinyl, examples of 5-membered heterocycloalkyl include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidinyl, imidazolidinyl, examples of tetrahydropyrazolyl, 6-membered heterocycloalkyl include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1, 4-thialkyl, 1, 4-dioxanyl, thiomorpholinyl, 1, 3-dithianyl, 1, 4-dithianyl, and examples of 7-membered heterocycloalkyl include, but are not limited to, azepanyl, oxepanyl, thiepanyl.

The term "cycloalkenyl" refers to a non-aromatic mono-or polycyclic ring system containing at least one carbon-carbon double bond. In some embodiments, the cycloalkenyl ring contains, for example, 3-10 ring atoms, 5-10 ring atoms, or 5-7 ring atoms. Non-limiting examples of suitable monocyclic cycloalkenyls include cyclopentenyl, cyclohexenyl, cycloheptenyl, and the like. A non-limiting example of a suitable polycyclocycloalkenyl group is norbornenyl.

The term "heterocycloalkenyl" refers to a non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond or carbon-nitrogen double bond and consisting solely of carbon and hydrogen atoms, wherein one or more ring atoms in the ring system is an element other than carbon, e.g., 1,2,3,4, 5 ring atoms are each independently a heteroatom selected from nitrogen, oxygen, and sulfur. There are no adjacent oxygen and/or sulfur atoms in the ring system, and in some embodiments, the heterocycloalkenyl ring contains, for example, 5 to 10 ring atoms, 5 to 8 ring atoms, 5 to 7 ring atoms, or, 5 to 6 ring atoms. The pre-conjugated aza, oxa or thia before the name heterocyclenyl indicates that at least one nitrogen, oxygen or sulfur atom is present as a ring atom, respectively. Non-limiting examples of suitable monocyclic azacycloalkenyls include 1,2,3,4 tetrahydropyridine, 1, 2-dihydropyridine, 1, 4-dihydropyridine, 1,2,3, 6-tetrahydropyridine, 1,4, 5, 6-tetrahydropyrimidine, 2-pyrrolidinyl, 3-pyrrolidinyl, 2-imidazolinyl, 2-pyrazolinyl, and the like. Non-limiting examples of suitable oxacycloalkenyl groups include 3, 4-dihydro-2H-pyran, dihydrofuranyl, fluorodihydrofuranyl, and the like. A non-limiting example of a suitable polycyclic oxacycloalkenyl group is 7-oxabicyclo [2.2.1] heptenyl.

The term "cycloalkynyl" refers to a non-aromatic monocyclic or polycyclic ring having at least one carbon-carbon triple bond and consisting of only carbon and hydrogen atoms, for example, a 4-15, 5-15, 6-10, 7-10 or 8-10 membered ring, such as an 8-to 10-membered monocyclic or 12-to 15-membered bicyclic ring. It may comprise one or more fused or bridged rings. Unless otherwise specified, the cycloalkynyl ring may be attached at any carbon atom which results in a stable structure, and, if substituted, may be substituted at any suitable carbon atom which results in a stable structure. Exemplary cycloalkynyl groups include cyclooctynyl, cyclononynyl, cyclodecynyl, 2-methylcyclooctynyl, and the like.

The term "heterocycloalkynyl" refers to a heterocycloalkyl group in which at least one carbon-carbon single bond is replaced by a carbon-carbon triple bond.

The term "aryl" or "aromatic ring" refers to an aromatic ring or an aromatic or partially aromatic ring system composed of carbon atoms and hydrogen atoms. It may be a single ring or may be multiple rings (e.g., more than 2 rings such as bicyclic rings) that are fused together or linked covalently. Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, and 1,2,3, 4-tetrahydronaphthalene, and the like. Depending on the structure, the aryl group may be a monovalent group or a divalent group, i.e., an arylene group.

The term "C₆-C₁₈Aryl "or" aromatic ring "refers to an aryl or aromatic ring as defined above having 6 to 18 carbon atoms (e.g., 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 carbon atoms).

"heteroaryl" or "heteroaromatic ring" refers to an aromatic ring group consisting of carbon atoms and at least one heteroatom (e.g., 1 to 5, such as 1,2,3,4, 5) selected from nitrogen, oxygen, and sulfur. The heteroaryl group may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may comprise fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl group may optionally be oxidized; the nitrogen atoms may optionally be quaternized. Examples include, but are not limited to, azanyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzothiadiazolyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothiophenyl (benzothiophenyl), benzotriazolyl, benzo [4,6] imidazo [1,2-a ] pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, furanyl, thienyl, furanonyl, isothiazolyl, imidazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, indolinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazatrolyl, oxazolyl, oxiranyl, phenazinyl, phenothiazinyl, phenoxazinyl, and benzoxazinyl, 2, 3-naphthyridinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl.

The term "C₁-C₁₈Heteroaryl "refers to an aromatic cyclic group having, as ring-forming atoms, at least one heteroatom (e.g., 1,2,3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 heteroatoms) selected from N, O, and S in addition to 1 to 18 carbon atoms (e.g., 1,2,3,4, 5 heteroatoms).

Unless otherwise indicated, the term "heterocyclyl" or "heterocycle" refers to a cyclic structure that may be saturated or unsaturated, aromatic or non-aromatic, wherein the cyclic structure contains at least one carbon and at least one heteroatom selected from O, N, S, examples of which include heterocycloalkyl, heteroaryl, heterocycloalkenyl, and heterocycloalkynyl.

The term "C₁-C₁₈The heterocyclic group "means a heterocyclic group as defined above having 1 to 18 carbon atoms (e.g., 1,2,3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 carbon atoms) and further containing at least one hetero atom (e.g., 1,2,3,4 or 5 hetero atoms) selected from N, O, and S as a ring-forming atom.

The term "carbocycle" refers to an aromatic or nonaromatic (partially or fully saturated) carbocycle, such as C3-7 carbocycle, C5-7 carbocycle, C5-12 carbocycle, C5-10 monocyclic carbocycle, and the like, examples of which include cycloalkyl, cycloalkenyl, cycloalkynyl, and aryl, such as: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclobutene, cyclopentene, cyclohexene, cycloheptene, cyclobutanediene, cyclopentadiene, cyclohexadiene, cycloheptadiene, or a benzene ring.

The term "4-7 membered nitrogen containing saturated heterocyclic ring" means a ring containing 1-5 (e.g., 1,2,3,4 or 5) heteroatoms selected from oxygen, nitrogen and sulfur, and necessarily containing one or more nitrogen atoms in a partially or fully saturated 4-7 membered monocyclic heterocyclic ring. For example: azetidine, pyrroline, pyrrolidine, imidazoline, imidazolidine, triazoline, triazolidine, tetrazoline, tetrazolidine, pyrazoline, pyrazolidine, dihydropyridine, tetrahydropyridine, piperidine, dihydropyrazine, tetrahydropyrazine, piperazine, dihydropyrimidine, tetrahydropyrimidine, perhydropyrimidine, dihydropyridazine, tetrahydropyridazine, perhydropyridazine, dihydroazepine, tetrahydroazepine, perhydroazepine, dihydrodiazepine, tetrahydrodiazepine, perhydrodiazepine, dihydrooxazole, tetrahydrooxazole (oxazolidine), dihydroisoxazole, tetrahydroisoxazole (isoxazole), dihydrothiazole, thiazolidine (thiazolidine), dihydroisothiazole, tetrahydroisothiazole (isothiazoline), dihydrofurazan, tetrahydrofurfurazan, dihydrooxadiazole, tetrahydrodiazole (diazolidine), dihydroazazine, tetraoxadiazine, dihydrooxadiazine, tetrahydroxadiazine, tetrahydroxadiazepine, tetrahydroxazine, dihydrooxadiazine, tetrahydrooxaazapine, tetrahydrooxazaazepine, perhydrooxazapine, dihydrooxadiazepine, pyrazoline, dihydrooxadiazine, dihydroazazole, dihydrooxadiazine, and pyrazoline, Tetrahydrooxadiaza, perhydrooxadiaza, dihydrothiadiazoles, tetrahydrothiadiazoles (thiadiazolidines), dihydrothiazines, tetrahydrothiazines, dihydrothiadiazines, tetrahydrothiadiazines, dihydrothiazazepines, tetrahydrothiazapines, perhydrothiazapines, dihydrothiadiaza, tetrahydrothiadiaza, perhydrothiadiaza, morpholine or thiomorpholine rings.

5-12 membered ring groups or 5-12 membered rings refer to C5-12 carbocyclic and 5-12 membered heterocyclic rings. 5-7 membered ring groups refer to C5-7 carbocyclic and 5-7 membered heterocyclic rings. Here, the carbocycle and heterocycle have the same meanings as above, and as the 5-to 7-membered heterocycle, for example: pyrroline, pyrrolidine, imidazoline, imidazolidine, triazoline, triazolidine, tetrazoline, pyrazoline, pyrazolidine, dihydropyridine, tetrahydropyridine, piperidine, dihydropyrazine, tetrahydropyrazine, piperazine, dihydropyrimidine, tetrahydropyrimidine, perhydropyrimidine, dihydropyridazine, tetrahydropyridazine, perhydropyridazine, dihydroazepine, tetrahydroazepine, perhydroazepine, dihydrodiaza, tetrahydrodiaza, perhydrodiaza, dihydrofuran, tetrahydrofuran, dihydropyran, tetrahydropyran, dihydrooxa, tetrahydrooxa, perhydrooxa, dihydrothiophene, tetrahydrothiophene, dihydrothiopyran, tetrahydrothiopyran, dihydrothia, tetrahydrothia, perhydrothia, dihydrooxazole, tetrahydrooxazole (oxazolidine), dihydroisoxazole, tetrahydroisoxazole (isoxazole), dihydrothiazole, thiazolidine (thiazolidine), dihydroisothiazole, tetrahydroisothiazole (isothiazole) thiazolidine, Dihydrofurazan, tetrahydrofurazan, dihydrodiazole, tetrahydrodiazole, dihydrooxazine, tetrahydrodiazine, dihydrodiazine, dihydroazepine, tetrahydroazepine, perhydroazepine, dihydrooxadiazide, tetrahydrooxadiazide, dihydrothiadiazole, tetrahydrothiadiazole, dihydrothiazine, tetrahydrothiazine, dihydrothiadiazine, tetrahydrothiadiazine, dihydrothiazazepine, tetrahydrothiazapine, perhydrothiazapine, dihydrothiadiazepine, tetrahydrothiadiazepine, perhydrothiadiazepine, morpholine, thiomorpholine, thiazine, dioxolane, dioxane, dithiolane, dithiane, pyrrole, imidazole, triazole, tetrazole, pyrazole, pyridine, piperazine, pyrimidine, pyridazine, aza, diazepine, furan, pyran, oxa, thiophene, thiopyran, thiaza, oxazole, isoxazole, thiazole, isothiazole, furazan, dihydrodiazine, dihydrothiadiazine, dihydrooxathiadiazine, dihydrooxathiadiazole, dihydrooxadiazepine, dihydrothiadiazole, dihydrothiadiazine, dihydrothiadiazole, dihydrothiadiazine, thiadiazole, dihydrothiadiazole, thiadiazole, and the use of a, Diazoles, oxazines, diazines, thiadiazoles, thiazines, thiadiazines, thiazepines or thiadiazacyclos.

In the context of the present specification,

meaning the attachment location.

Double lines formed by dotted lines and solid lines in this specification

Represents a single bond or a double bond.

The term "carboxyalkyl" refers to a group having the structure-alkyl-COOH, wherein alkyl is as defined above. Representative examples of carboxyalkyl groups include, but are not limited to, carboxymethyl (-CH)₂CO₂H) 2-carboxyethyl and the like. "optionally substituted carboxyalkyl" means that the alkyl portion of the group is substituted or unsubstituted.

The term "optionally substituted amino" refers to-NH₂Mono-or di-substituted amino, and 5-to 7-membered cyclic amino.

The term "ester group" as used herein refers to a group having the structure-c (o) OR ' oc (o) R ', wherein R ' is optionally substituted alkyl, optionally substituted aryl OR optionally substituted heteroaryl as defined above. In some embodiments, R' is alkyl, haloalkyl, aryl, heteroaryl, alkaryl, alkylheteroaryl. In some embodiments, R' is alkyl or haloalkyl. In some embodiments, R' is C_1-6An alkyl group. By "optionally substituted ester group" is meant that the R' moiety in the group is substituted or unsubstituted.

The term "acyl" as used herein refers to a group having the structure-C (O) R ', wherein R' is as defined above. "optionally substituted acyl" means that the R' moiety in the group is substituted or unsubstituted.

The term "sulfonyl" as used herein means having an-SO₂A group of the structure R ', wherein R' is as defined above. "optionally substituted sulfonyl" means that the R' moiety in the group is substituted or unsubstituted.

The term "optionally substituted" means that the group is unsubstituted or substituted with one or more substituents (e.g., 1 to 4, 1 to 3, or 1 to 2) which, when substituted, are one or more groups individually and independently selected from the group consisting of: halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, alkoxy, aryloxy, heteroaryloxy, mercapto, alkylthio, arylthio, cyano, carbonyl, thiocarbonyl, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, S-sulfonamido, N-sulfonamido, carboxy, C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, nitro, ester, silyl, trihalomethanesulfonyl, or amino groups including mono-and di-substituted amino groups, protected derivatives thereof, and the like. Whenever a substituent is described as being "optionally substituted", the substituent may be substituted with one of the substituents described above.

When a group is substituted with more than one substituent, the substituents may be the same or different, any substituted functional group herein may be substituted at 1 to 4 different positions, and those 1 to 4 substituent groups can each be independently substituted at 1 to 4 positions.

The term "pharmaceutically acceptable salt" refers to salts that retain the biological potency of the free acid and free base of the specified compound, and that are biologically or otherwise not adversely affected. "pharmaceutically acceptable salts" include "pharmaceutically acceptable acid addition salts" and "pharmaceutically acceptable base addition salts".

The compounds of the present application may exist as specific tautomers, and all such isomers and mixtures thereof are included within the scope of the present application.

The present application also includes isotopically-labeled compounds of the present application, which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine and chlorine, such as respectively²H、³H、¹¹C、¹³C、¹⁴C、¹³N、¹⁵N、¹⁵O、¹⁷O、¹⁸O、³¹P、³²P、³⁵S、¹⁸F、¹²³I、¹²⁵I and³⁶cl, and the like.

Certain isotopically-labelled compounds of the present application (e.g. with³H and¹⁴c-labeled ones) can be used in compound and/or substrate tissue distribution assays. Tritiated (i.e. by tritiation)³H) And carbon-14 (i.e.¹⁴C) Isotopes are particularly preferred for their ease of preparation and detectability. Positron emitting isotopes, such as¹⁵O、¹³N、¹¹C and¹⁸f can be used in Positron Emission Tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present application can generally be prepared by following procedures analogous to those disclosed in the schemes and/or in the examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

In addition, heavier isotopes are used (such as deuterium (i.e., deuterium)²H) Substitution may provide certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements), and thus may be preferred in certain circumstances, wherein deuterium substitution may be partial or complete, partial deuterium substitution meaning that at least one hydrogen is substituted with at least one deuterium, all such forms of the compounds being encompassed within the scope of the present application.

In the present invention, the term "subject" includes humans and animals, for example, mammals (e.g., primates, cows, horses, pigs, dogs, cats, mice, rats, rabbits, goats, sheep, and birds, etc.).

The term "treating" means administering a compound or formulation described herein to ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:

(i) inhibiting the disease or disease state, i.e., arresting its development;

(ii) alleviating the disease or condition, i.e., causing regression of the disease or condition.

The term "preventing" means administering a compound or formulation described herein to prevent a disease or one or more symptoms associated with the disease, and includes: prevention of a disease or condition occurs in a mammal, particularly when such mammal is susceptible to the disease condition, but has not yet been diagnosed as having the disease condition.

The term "effective amount," or "therapeutically effective amount," refers to an amount of at least one agent or compound that is sufficient to alleviate one or more symptoms of the disease or disorder being treated to some extent after administration. The result may be a reduction and/or alleviation of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is clinically necessary to provide a significant remission effect of the condition. An effective amount suitable in any individual case can be determined using techniques such as a dose escalation assay.

The term "composition" or "pharmaceutical composition" refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical ingredient including, but not limited to, carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and/or excipients.

"pharmaceutically acceptable adjuvants" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that has been recognized by the U.S. food and drug administration as being useful in humans or animals in a variety of forms that do not have any adverse effects on the resulting pharmaceutical composition.

The pharmaceutical compositions of the present application can be prepared by combining the compounds of the present application with suitable pharmaceutically acceptable excipients, for example, can be formulated into solid, semi-solid, liquid or gaseous formulations such as tablets, pills, capsules, powders, granules, lozenges, ointments, syrups, emulsions, suspensions, solutions, suppositories, injections, inhalants, gels, microspheres, aerosols, and the like.

Typical routes of administration or administration of the compounds of the present application, pharmaceutically acceptable salts, tautomers or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, transmucosal, topical, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

The compounds or compositions of the present invention may be formulated and used as the following dosage forms: tablets, capsules or elixirs for oral administration; suppositories for rectal administration; sterile solutions for administration by injection, suspensions; patches for transdermal administration, subcutaneous deposits, and the like. Injections can be prepared in the following conventional forms: solutions or suspensions, solid dosage forms suitable for constitution with a solution or suspension prior to injection, or emulsions. Suitable excipients are, for example, water, saline, dextrose, mannitol, lactose, lecithin, albumin, sodium glutamate, cysteine hydrochloride and the like. In addition, if desired, the injectable pharmaceutical compositions may contain minor amounts of non-toxic adjuvants such as wetting agents, pH buffering agents and the like. Absorption enhancers (e.g., liposomes) can also be used if desired.

Formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Alternatively, suspensions of the active compounds may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or other organic oils such as soybean oil, grapefruit oil or almond oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, for example sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, the suspension may contain suitable stabilizers or agents that increase the solubility of the compound, allowing for the preparation of highly concentrated solutions.

The pharmaceutical compositions of the present application can be prepared by methods known in the art, such as conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, and lyophilizing processes.

In some embodiments, the pharmaceutical compositions of the present application are in oral form. For oral administration, the pharmaceutical compositions may be formulated by mixing the active compound with pharmaceutically acceptable adjuvants or excipients well known in the art. These adjuvants or excipients enable the compounds of the present application to be formulated as tablets, pills, dragees, capsules, powders, granules, liquids, syrups, emulsions, gels, slurries, suspensions and the like, for oral administration to a patient.

Solid pharmaceutical compositions suitable for oral administration may be prepared by conventional mixing, filling or tableting methods. For example, oral compositions in solid form can be obtained by: the active compound is mixed with solid adjuvants or excipients, the mixture obtained is optionally milled, if desired with further suitable adjuvants or excipients, and the mixture is then processed to granules, to give tablets or dragee cores. Suitable adjuvants or excipients include, but are not limited to: fillers, binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents and the like. Pharmaceutical preparations for oral use, for example, can be obtained by the following process: combining the active compound with a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, if desired after addition of suitable auxiliaries, to give tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, for example cross-linked polyvinylpyrrolidone, agar or alginic acid or alginates such as sodium alginate. Sugar-coated cores are suitably coated. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablets or dragee coatings in order to identify or characterize different combinations of active compound doses. These formulations can be made using methods well known in the art.

In all methods of administration of the compounds or compositions described herein, the daily dose administered may be, for example, from 0.001 to 300mg/kg body weight, such as from 0.01 to 300mg/kg body weight, or from 10 to 200mg/kg body weight, administered as a single dose or as divided doses.

The term "fluorouracil-based drug" includes, but is not limited to, fluorouracil or derivatives thereof, or drugs containing fluorouracil structures, for example, 5-fluorouracil (5-FU), 5'-2' -deoxyuridine, tegafur (tegafur), fluorouracil, carmofur, deoxyfluorouridine, tegafur/uracil, capecitabine (capecitabine), tegafur, medusmidine, pyrifudin, flutriafolan, and cisdara, and the like.

The expression "a disease, disorder or condition that benefits from inhibition of DPD expression" includes, but is not limited to: digestive tract tumor, lung cancer, breast cancer, metastatic non-small cell lung cancer, metastatic breast cancer, metastatic pancreatic cancer, metastatic bile duct cancer, head and neck tumor (such as oral cancer), middle and late stage nasopharyngeal carcinoma, etc.

wherein:

R₂selected from hydrogen or optionally substituted alkyl;

ring B is an aromatic or heteroaromatic ring;

x is an integer from 1 to 2; and

y is an integer of 1,2 or 3.

In some embodiments, R₁Selected from aryl, heteroaryl, or heterocycloalkenyl each independently optionally substituted with halogen, -OH, -CN, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl, alkoxy, aryloxy, or heteroaryloxy;

in some embodiments, R₁Selected from the group consisting of independently optionally substituted halogen, -OH, -CN, C_1-6Alkyl radical, C_1-6Haloalkyl, C_1-6Hydroxyalkyl, 3-to 10-membered cycloalkyl, C_1-6Alkoxy radical, C₆-C₁₈Aryloxy radical or C₁-C₁₈Heteroaryloxy-substituted C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl, or 5-10 membered heterocycloalkenyl;

in some embodiments, R₁Selected from the group consisting of independently optionally substituted halogen, -OH, -CN, C_1-6Alkyl radical, C_1-6Alkoxy or C₆-C₁₈Aryloxy substituted C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl, or 5-10 membered heterocycloalkenyl;

in some embodiments, R₁Selected from C, each independently optionally substituted with fluoro, chloro, -OH, -CN, methyl, ethyl, n-propyl, isopropyl, n-propoxy, isopropoxy or phenoxy₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl, or 5-10 membered heterocycloalkenyl;

in some embodiments, R₁Selected from C, each independently optionally substituted with fluoro, -OH, -CN, methyl, n-propoxy, isopropoxy or phenoxy₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl, or 5-10 membered heterocycleAn alkenyl group;

in some embodiments, R₁Selected from: optionally selected from halogen, -OH, -CN, C_1-6Alkyl radical, C_1-6Alkoxy, or C₆-C₁₈C substituted by aryloxy radicals₆-C₁₈An aryl group; optionally is covered with C_1-6Alkoxy-substituted C₁-C₁₈A heteroaryl group; or 5-10 membered heterocycloalkenyl;

in some embodiments, R₁Said C occurring in the definition of₆-C₁₈Aryl is phenyl;

in some embodiments, R₁Said C occurring in the definition of₁-C₁₈Heteroaryl is selected from pyridyl or furyl;

in some embodiments, the 5-10 membered heterocycloalkenyl is

In some embodiments, the halogen is selected from fluorine or chlorine;

in some embodiments, R₁Selected from: phenyl optionally substituted with a group selected from fluoro, -OH, -CN, methyl, n-propoxy, or phenoxy; an isopropoxy substituted pyridyl group; a furyl group; or

In some embodiments, R₂Selected from hydrogen, optionally substituted C_1-6An alkyl group;

in some embodiments, R₂Selected from optionally substituted C_1-6An alkyl group;

in some embodiments, R₂Selected from C optionally substituted by halogen, alkenyl or hydroxy_1-6An alkyl group;

in some embodiments, R₂Selected from C optionally substituted by alkenyl or hydroxy_1-6An alkyl group;

in some embodiments, R₂Is methyl, ethyl, n-propyl, isopropyl, allylRadical or-CH₂CH₂CH₂OH；

In some embodiments, R₂Is methyl, allyl or-CH₂CH₂CH₂OH；

In some embodiments, R₂Is methyl.

In some embodiments, each R is₃Independently selected from hydrogen, deuterium, halogen, -CN, -NO₂Nitroso group, carboxyl group, sulfonic acid group, optionally substituted sulfonyl group, optionally substituted C_1-6Alkyl, optionally substituted C_3-8Cycloalkyl, optionally substituted C₆-C₁₈Aryl, optionally substituted C₁-C₁₈A heteroaryl group;

in some embodiments, each R is₃Independently selected from: hydrogen; and each independently optionally halogen, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl or C_1-6Alkoxy-substituted C_1-6Alkyl radical, C_3-8Cycloalkyl radical, C₆-C₁₈Aryl or C₁-C₁₈A heteroaryl group;

in some embodiments, each R is₃Independently selected from: hydrogen; c_3-8A cycloalkyl group; optionally substituted by halogen, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl or C_1-6Alkoxy-substituted C_1-6An alkyl group; each independently optionally substituted by halogen, or C_1-6Alkoxy-substituted C₆-C₁₈Aryl or C₁-C₁₈A heteroaryl group;

in some embodiments, each R is₃Independently selected from: hydrogen; c_3-8A cycloalkyl group; optionally substituted by halogen or C₆-C₁₈Aryl substituted C_1-6An alkyl group; optionally substituted by halogen, or C_1-6Alkoxy-substituted C₆-C₁₈An aryl group; or C optionally substituted by halogen₁-C₁₈A heteroaryl group;

in some embodiments, each R is₃Independently selected from: hydrogen; c_3-8A cycloalkyl group; c optionally substituted by halogen or phenyl_1-6An alkyl group; renIs selected from halogen, or C_1-6Alkoxy-substituted phenyl; or furyl or thienyl each independently optionally substituted with halogen;

in some embodiments, each R is₃Independently selected from hydrogen, methyl, n-propyl, isopropyl, cyclopropyl, cyclohexyl, -CF₃Benzyl, phenyl, fluoro-substituted phenyl, methoxy-substituted phenyl, furyl or chloro-substituted thienyl;

in some embodiments, each R is₃Independently selected from hydrogen, or methyl;

in some embodiments, at least one R is₃Is not hydrogen and is para to N;

in some embodiments, x is 1.

In some embodiments, R₄、R₅And R₆Each independently selected from: hydrogen; or an alkyl group optionally substituted with a group selected from a hydroxyl group, an optionally substituted ester group, an optionally substituted aminocarbonyl group, or a carboxyl group;

in some embodiments, R₄、R₅And R₆Each independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, optionally substituted ester group, optionally substituted aminocarbonyl, or carboxy_1-6An alkyl group;

in some embodiments, R₄、R₅And R₆Each independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl, or carboxy_1-6An alkyl group;

in some embodiments, R₄、R₅And R₆Each independently selected from: hydrogen, methyl, ethyl, -CH₂COOH、-CH₂CH₂OH、-CH₂CH₂CH₂OH、-CH₂CONH₂Or CH₂COOEt；

In some embodiments, R₄、R₅And R₆Each independently selected from hydrogen or methyl;

in some embodiments, R₄、R₅And R₆Are all hydrogen.

In some embodiments, the heteroaromatic ring present in the definition of ring B is C₁-C₁₈A heteroaromatic ring;

in some embodiments, the heteroaryl ring present in the definition of ring B is a 5-10 membered heteroaryl ring;

in some embodiments, the aromatic ring present in the definition of ring B is C₆-C₁₈An aromatic ring;

in some embodiments, the aromatic ring present in the definition of ring B is C₆-C₁₀An aromatic ring;

in some embodiments, the aromatic ring present in the definition of ring B is a benzene ring;

in some embodiments, ring B is a heteroaryl ring;

in some embodiments, the heteroaromatic ring present in the definition of ring B is pyridine or thiophene;

in some embodiments, the heteroaromatic ring present in the definition of ring B is pyridine;

in some embodiments, the heteroaryl ring present in the definition of ring B is optionally substituted with 1-2R in the ortho position to the heteroatom₇Substituted by groups;

in some embodiments of the present invention, the substrate is,

selected from pyridyl or thienyl optionally substituted with: hydrogen, fluorine, chlorine, bromine, trifluoromethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH₂CH₂OCH₃，-OCH₂CH(OH)CH₂OH, N-ethyl-amino or N-ethyl-N-methyl-amino;

in some embodiments of the present invention, the substrate is,

selected from 4-substituted 2, 6-dichloropyridine, 4-substituted 2, 6-dibromopyridine, 4-substituted 2-chloro-6-hydroxyethylpyridine and 4-substituted-2-chloro-6-hydroxypropylPicoline, 4-substituted 2-chloro-6-ethoxypyridine, 4-substituted-2-chloro-6-n-propoxypyridine, 4-substituted-2-chloro-6-isopropoxypyridine, 4-substituted-2-chloro-6-n-butoxypyridine, 4-substituted-2-chloro-6- (2-methoxyethoxy) pyridine, 4-substituted-2-trifluoromethyl-6-ethoxypyridine, 4-substituted-2-trifluoromethyl-6- (2, 3-dihydroxypropoxy) pyridine, and a salt thereof, 4-substituted-2- (N-ethylamino) -6-chloropyridine, 4-substituted-2- (N-ethyl-N-methylamino) -6-chloropyridine, or 5-substituted-2, 3-dichlorothiophene;

in some embodiments of the present invention, the substrate is,

is 4-substituted 2, 6-dichloropyridine.

In some embodiments, each R is₇Independently selected from hydrogen; halogen; alkyl optionally substituted with halogen, hydroxy or alkoxy; alkoxy optionally substituted with halogen, hydroxy or alkoxy; or amino optionally substituted with alkyl;

in some embodiments, each R is₇Independently selected from hydrogen; halogen; optionally substituted by halogen, hydroxy or C_1-6Alkoxy-substituted C_1-6An alkyl group; optionally substituted by halogen, hydroxy or C_1-6Alkoxy-substituted C_1-6An alkoxy group; or optionally is covered with C_1-6An alkyl-substituted amino group;

in some embodiments, each R is₇Independently selected from hydrogen; halogen; c optionally substituted by halogen or hydroxy_1-6An alkyl group; optionally substituted by hydroxy or C_1-6Alkoxy-substituted C_1-6An alkoxy group; or optionally is covered with C_1-6An alkyl-substituted amino group;

in some embodiments, each R is₇Independently selected from hydrogen; halogen; c optionally substituted by halogen_1-6An alkyl group; optionally substituted by hydroxy or C_1-6Alkoxy-substituted C_1-6An alkoxy group;

in some embodiments, each R is₇Independently selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, methoxy, ethoxy, n-propoxy, iso-propoxyPropoxy, n-butoxy, -OCH₂CH₂OCH₃，-OCH₂CH(OH)CH₂OH, N-ethyl-amino or N-ethyl-N-methyl-amino;

in some embodiments, each R is₇Independently selected from hydrogen or chlorine.

in a third aspect, the present application provides a process for the preparation of a compound of the first aspect, comprising:

step 1:

step 2:

and the following step 3-1 or step 3-2:

step 3-1:

step 3-2:

In some embodiments, when R₆Selected from hydrogen, the method comprises any of step 3-1 or step 3-2.

In some embodiments, when R₆When selected from hydrogen, the process comprises step 3-1; and when R is₆When selected from optionally substituted alkyl groups, the process comprises step 3-2.

In some embodiments, step 1 is performed in the presence of a catalyst. In some embodiments, step 1 is performed in the presence of a catalyst selected from the group consisting of: pd (PPh)₃)₄、PdCl₂(PPh₃)₂、PdCl₂(PhCN)₂、Pd(OAc)₂、Pd/C、PdCl₂(dppf), combinations thereof or the like; in some embodiments, the catalyst in step 1 is PdCl₂(dppf)。

In some embodiments, step 1 is performed in the presence of a base. In some embodiments, step 1 is performed in the presence of a base selected from the group consisting of: c_SF、K₂CO₃、K₃PO₄Or combinations thereof, and the like; in some embodiments, the base in step 1 is cesium fluoride.

In some embodiments, step 1 is performed in the presence of a solvent; in some embodiments, the solvent in step 1 is selected from 1, 4-dioxane, a mixture of toluene and water, a mixture of ethanol and water, or other Suzuki reacted solvents;

in some embodiments, step 1 is performed at room temperature or under heated conditions, such as at a temperature of from room temperature to 120 ℃ (e.g., from room temperature to 100 ℃, from room temperature to 80 ℃).

In some embodiments, step 3-2 is performed in the presence of a base; in some embodiments, step 3-2 is performed at a time selected from EtN (i-Pr)₂In the presence of a base.

In some embodiments, step 3-2 is performed in the presence of a solvent; in some embodiments, said step 3-2 is carried out in the presence of a solvent selected from DCM (dichloromethane).

In some embodiments, step 3-2 is performed at 0 ℃.

In some embodiments, the method further comprises the following step 1 a:

step 1 a:

in some embodiments, wherein R is₀' is I, step 1a in NIS and HBF₄In the presence of (a). In some embodiments, R₀Is Br.

In some embodiments, step 1a is performed in the presence of a solvent; in some embodiments, the solvent in step 1a is selected from CH₃Aprotic solvents miscible with water, such as CN, DMF, DMSO or THF; in some embodiments, step 1a is performed at room temperature or under heated conditions, for example at a temperature of 80 ℃.

In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

In some embodiments, the pharmaceutical composition further comprises one or more additional therapeutic agents.

In some embodiments, the additional therapeutic agent is an anti-tumor drug.

In some embodiments, the anti-neoplastic drug is selected from fluorouracil drugs.

In some embodiments, the antineoplastic agent is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouracil, carmofur, doxifluridine, uracil tegafur, capecitabine, tegafur, Youfodin, pyrifudin, flutolterone, and voroda.

In some embodiments, the disease, disorder or condition that benefits from inhibition of DPD expression is selected from a tumor.

In some embodiments, the disease, disorder or condition or the tumor is selected from a tumor of the digestive tract, a tumor of the head and neck, lung cancer, breast cancer, or the like; optionally, the head and neck tumor comprises oral cancer and the like;

in some embodiments, the tumor is a tumor that is resistant to fluorouracil.

In some embodiments, the one or more additional therapeutic agents are anti-neoplastic agents.

In some embodiments, the fluorouracil drug is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouracil, carmofur, doxifluridine, uracil tegafur, capecitabine, tegafur, Youfodin, pyrifudin, flutolterone, and voroda.

in some embodiments, the disease, disorder or condition that benefits from inhibition of DPD expression is selected from tumors that are resistant to fluorouracil drugs.

In some embodiments, the fluorouracil drug is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouracil, carmofur, doxifluridine, tegafur/uracil, capecitabine, tegafur, Youfuding, pyrifudin, flutolterone, and voroda.

In some embodiments, the S1PR2 inhibitor is a compound having a structure represented by formula II or formula III, or a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition thereof, or a combination thereof with one or more other therapeutic agents:

in said formula II:

Ar¹is an optionally substituted non-aromatic heterocyclic group, an optionally substituted aryl group or an optionally substituted heteroaryl group;

Ar²is an optionally substituted non-aromatic heterocyclic group, or an optionally substituted heteroaryl group;

w is-NR^a—、O、—CO—、—CONH—、—NHCH₂-or-CR^bR^c—；

Z is-C (═ O) -, -C (═ S) -, O, — CR^bR^c—、═N—、═CH—、

Y is-NR^a—、—C(═O)—、O、—CR^bR^c-N ═, -CH ═, ═ N-or ═ CH-; and

x is a direct bond, -NR^a-, -N ═, -C (═ O) -, -CH ═, O or-CR^bR^c—；

Wherein R is^a、R^bAnd R^cEach independently selected from H or optionally substituted alkyl.

In said formula III:

R¹represents (1) optionally substituted alkyl, (2) optionally substituted alkenyl, (3) optionally substituted alkynyl, (4) optionally substituted C_3-7Carbocyclic ring, or (5) -CONR³¹R³²；

R³¹And R³²Each independently represents (1) a hydrogen atom, or (2) an optionally substituted alkyl group;

R²represents (1) a hydrogen atom, (2) an optionally substituted alkyl group;

R³and R⁴Each independently represents (1) a halogen atom, (2) an optionally substituted alkyl group, (3) an optionally substituted alkoxy group, (4) a hydroxyl group, (5) -L-CONR⁶R⁷，(6)-L-SO₂R⁸Or (7) -L-COOR⁹；

R⁵Represents (1) a hydrogen atom, (2) a halogen atom, (3) an optionally substituted alkyl group;

l represents (1) a chemical bond, (2) a group represented by the following formula 2, (3) an alkenylene group, (4) -O-alkenylene group, (5) an oxygen atom, or (6) a nitrogen atom optionally substituted with an alkyl group,

[ formula 2 ]]

In formula 2, A represents (1) a chemical bond, or (2) an oxygen atom; r¹²And R¹³Each independently represents (1) a hydrogen atom(2) optionally substituted alkyl, (3) hydroxy, or (4) NH₂Or (5) R¹²And R¹³Together with the carbon atom to which they are bonded form C_3-7A carbocyclic ring; the right arrow is bonded to-CONR⁶R⁷、-SO₂R⁸or-COOR⁹，

R⁶And R⁷Each independently represents (1) a hydrogen atom, (2) an optionally substituted alkyl group, (3) a hydroxyl group, (4) -CONR¹⁵R¹⁶，(5)-SO₂NR¹⁵R¹⁶，(6)-COR¹⁷Or (7) -SO₂R¹⁷Or R is⁶And R⁷Together with the nitrogen atom to which they are bonded form a 4-7 membered nitrogen containing saturated heterocyclic ring optionally substituted with hydroxy;

R⁸represents (1) optionally substituted alkyl, or (2) NR¹⁰R¹¹；

R⁹Represents (1) a hydrogen atom, or (2) an optionally substituted alkyl group;

R¹⁰and R¹¹Each independently represents (1) a hydrogen atom, (2) an optionally substituted alkyl group, (3) -CONR¹⁵R¹⁶，(4)-SO₂NR¹⁵R¹⁶，(5)-COR¹⁷Or (6) -SO₂R¹⁷；

E represents optionally substituted alkyl or

Ring 1 and ring 2 each independently represent a 5-12 membered ring group;

R¹⁴represents (1) a hydrogen atom, or (2) a hydroxyl group;

R¹⁵and R¹⁶Each independently represents (1) a hydrogen atom, (2) an optionally substituted alkyl group, or (3) a 5-to 12-membered ring group;

R¹⁷represents (1) optionally substituted alkyl, or (2) a 5-12 membered ring group;

M¹and M²Each independently represents (1) a chemical bond, (2) -C (O) -, (3) -O-, (4) -S-, (5) -C (O) O-, (6) -CH₂O-, or (7) -C (O) NH-;

n represents an integer of 1 to 2;

m represents an integer of 1 to 2;

p represents an integer of 0 to 5;

q represents an integer of 0 to 5;

r represents an integer of 0 to 4;

t represents an integer of 1 to 4;

when p is 2 or more, plural R³May be the same or different;

when q is 2 or more, plural R⁴May be the same or different;

when R is 2 or more, plural R⁵May be the same or different;

when t is 2 or more, plural R¹²And R¹³May be the same or different, respectively.

In some embodiments, in said formula II:

Ar¹is an optionally substituted non-aromatic 5-18 membered heterocyclic group, optionally substituted C₆-C₁₈Aryl or optionally substituted C₁-C₁₈A heteroaryl group;

Ar²is an optionally substituted non-aromatic 5-18 membered heterocyclic group, or an optionally substituted C₁-C₁₈A heteroaryl group;

w is-NR^a—、O、—CO—、—CONH—、—NHCH₂-or-CR^bR^c—；

Z is-C (O) -, -C (S) -, C (S) -and C (S) -are independently substituted,

Y is-NR^a-, O, or-CR^bR^cA; and

x is a direct bond, -NR^a-, - (C) (O) -, O or-CR^bR^c—；

Wherein R is^a、R^bAnd R^cEach independently selected from H or optionally substituted C_1-6An alkyl group.

In some embodiments, in said formula III:

R¹represents (1) optionally substituted by 1-5R²¹Substituted C_1-8Alkyl, (2) optionally substituted with 1-5R²¹Substituted C_2-8Alkenyl, (3) optionally substituted with 1-5R²¹Substituted C_2-8Alkynyl, (4) optionally substituted with 1-5 substituents selected from C_1-4Alkyl radical, C_1-4Haloalkyl, C_1-4Alkoxy and halogen substituted C_3-7Carbocyclic ring, or (5) -CONR³¹R³²；

R²¹Represents (1) a halogen atom, (2) -OR²²，(3)-NR²³R²⁴Or (4) an oxo group, wherein,

R²²represents (1) a hydrogen atom, (2) C_1-4Alkyl or (3) C_1-4A halogenated alkyl group,

R²³and R²⁴Each independently represents (1) a hydrogen atom, or (2) C_1-4An alkyl group;

R³¹and R³²Each independently represents (1) a hydrogen atom, or (2) C_1-4An alkyl group;

R²represents (1) a hydrogen atom, (2) C_1-4Alkyl, or (3) C_1-4A haloalkyl group;

R³and R⁴Each independently represents (1) a halogen atom, (2) C_1-4Alkyl group, (3) C_1-4Haloalkyl, (4) optionally halogen-substituted C_1-4Alkoxy, (5) hydroxy, (6) -L-CONR⁶R⁷，(7)-L-SO₂R⁸Or (8) -L-COOR⁹；

R⁵Represents (1) a hydrogen atom, (2) a halogen atom, or (3) C_1-4Alkyl, or (4) C_1-4A haloalkyl group;

l represents (1) a chemical bond, (2) a group represented by the following formula 2, (3) C_2-4Alkenylene, (4) -O-C_2-4Alkenylene, (5) oxygen, or (6) optionally substituted by C_1-4A nitrogen atom substituted with an alkyl group,

[ formula 2 ]]

In formula 2, A tableRepresents (1) a chemical bond, or (2) an oxygen atom; r¹²And R¹³Each independently represents (1) a hydrogen atom, (2) C_1-4Alkyl, (3) hydroxy, or (4) NH₂Or (5) R¹²And R¹³Together with the carbon atom to which they are bonded form C_3-7A carbocyclic ring; the right arrow is bonded to-CONR⁶R⁷、-SO₂R⁸or-COOR⁹，

R⁶And R⁷Each independently represents (1) a hydrogen atom, (2) C_1-4Alkyl group, (3) C_1-4Haloalkyl, (4) hydroxy, (5) -CONR¹⁵R¹⁶，(6)-SO₂NR¹⁵R¹⁶，(7)-COR¹⁷Or (8) -SO₂R¹⁷Or R is⁶And R⁷Together with the nitrogen atom to which they are bonded form a 4-7 membered nitrogen containing saturated heterocyclic ring optionally substituted with hydroxy;

R⁸represents (1) C_1-4Alkyl group, (2) C_1-4Haloalkyl, or (3) NR¹⁰R¹¹；

R⁹Represents (1) a hydrogen atom, or (2) C_1-8An alkyl group;

R¹⁰and R¹¹Each independently represents (1) a hydrogen atom, (2) C_1-4Alkyl group, (3) -CONR¹⁵R¹⁶，(4)-SO₂NR¹⁵R¹⁶，(5)-COR¹⁷Or (6) -SO₂R¹⁷；

E represents alkyl optionally substituted by halogen or

Ring 1 and ring 2 each independently represent a 5-7 membered cyclic group;

R¹⁴represents (1) a hydrogen atom, or (2) a hydroxyl group;

R¹⁵and R¹⁶Each independently represents (1) a hydrogen atom, (2) C_1-4Alkyl, or (3) a 5-7 membered ring group;

R¹⁷represents (1) C_1-4Alkyl, or (2) a 5-7 membered ring group;

n represents an integer of 1 to 2;

m represents an integer of 1 to 2;

p represents an integer of 0 to 5;

q represents an integer of 0 to 5;

r represents an integer of 0 to 4;

t represents an integer of 1 to 4;

when p is 2 or more, plural R³May be the same or different;

when q is 2 or more, plural R⁴May be the same or different;

when R is 2 or more, plural R⁵May be the same or different;

In some embodiments, in said formula II:

Ar¹is optionally substituted aryl or optionally substituted heteroaryl;

in some embodiments, Ar¹Is aryl or heteroaryl, each independently optionally substituted with s groups independently selected from: halogen; optionally substituted alkyl; optionally substituted alkoxy; optionally substituted cycloalkyl; optionally substituted heterocycloalkyl; optionally substituted-O-cycloalkyl; optionally substituted-O-heterocycloalkyl; -SO₂-an alkyl group; -SO₂NR^15aR^15b；-C(＝O)NR^15cR^15d(ii) a An optionally substituted amino group; a hydroxyl group; a cyano group; and a carboxyl group; s represents 0, 1,2,3 or 4;

in some embodiments, Ar¹Is aryl or heteroaryl, each independently optionally substituted with s groups independently selected from: halogen; -C_1-6Alkyl optionally substituted by one or more independently selected R¹³Substituted by groups; -C_1-6Alkoxy optionally substituted by one or more independently selected R¹³Substituted by groups; -C_3-7Monocyclic cycloalkyl optionally independently selected by one or moreR of (A) to (B)¹⁴Substituted by groups; a 4 to 11 membered monocyclic or fused or spirobicyclic heterocycloalkyl containing one, two or three heteroatoms independently selected from N, O and S, optionally substituted with one or more independently selected R¹⁴Substituted by groups; -O-C_3-7Monocyclic cycloalkyl optionally substituted with one or more independently selected R¹⁴Substituted by groups; -O-heterocycloalkyl, wherein the heterocycloalkyl is a 4 to 7 membered monocyclic heterocycloalkyl containing one, two or three heteroatoms independently selected from N, O and S, optionally substituted with one or more independently selected R¹⁴Substituted by groups; -SO₂-C_1-6An alkyl group; -SO₂NR^15aR^15b；-C(＝O)NR^15cR^15d；-NR^17aR^17b(ii) a A hydroxyl group; a cyano group; and a carboxyl group;

wherein each R is¹³Independently selected from: halogen; -CN; -OH; -C_1-4Alkoxy optionally substituted by one or more independently selected OH, C_1-4Alkoxy or halogen substitution; -C (═ O) NR^16aR^16b；

-NR^16cC(＝O)-C_1-4An alkyl group; -NR^16dC(＝O)-C_1-4An alkoxy group; -SO₂-C_1-4An alkyl group; -SO₂NR^16eR^16f；-NR^16gSO₂-C_1-4An alkyl group; -C_3-7Monocyclic cycloalkyl optionally substituted with one or more independently selected halogen or C optionally substituted with one or more halogen_1-4Alkyl substitution; and 4 to 7 membered monocyclic heterocycloalkyl comprising one, two or three heteroatoms independently selected from N, O and S, optionally substituted with one or more independently selected halogen or C optionally substituted with one or more halogen_1-4Alkyl substitution;

each R¹⁴Independently selected from: halogen; -CN; -OH; -C_1-4Alkoxy, optionally substituted with one or more halogen; and-C_1-4Alkyl, optionally substituted with one or more halogens;

each R^15a、R^15b、R^15c、R^15d、R^16a、R^16b、R^16c、R^16d、R^16e、R^16fAnd R^16gIndependently selected from H and C_1-4An alkyl group;

each R^17aAnd R^17bIndependently selected from H and optionally one or more independently selected halogen, OH or C_1-4Alkoxy-substituted C_1-4An alkyl group.

In some embodiments, Ar²Is with Ar¹The same monocyclic heteroaryl group is represented by one of the following structures (1) to (3):

wherein in the structure (1):

R₁is hydrogen or optionally substituted alkyl;

R₂selected from hydrogen, deuterium, halogen, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, optionally substituted cycloalkyl or optionally substituted cycloalkoxy;

R₃and R₄Each independently selected from hydrogen, deuterium, halogen, -CN, -NO₂Nitroso, carboxyl, sulfonic acid, optionally substituted sulfonyl, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, optionally substituted cycloalkyl, optionally substituted cycloalkoxy, optionally substituted aryl or optionally substituted heteroaryl;

R₃and R₄Can be located at h, i or j, but cannot be located at the same position at the same time;

wherein in the structure (2):

a is a direct bond or (CR), B, C and D are each independently selected from (CR) and N, wherein R is H or optionally substituted alkyl; provided that B, C and D are not both N, and when A is a direct bond, D is (CR);

R³is optionally substituted alkyl;

x is selected from O, NR⁴And CR⁴R⁵Wherein R is⁴、R⁵Each independently selected from H, optionally substituted alkylSubstituted cycloalkyl, optionally substituted alkoxy, halogen, hydroxy, nitrile and carboxy;

y is selected from O or S; and

z is an optionally substituted aromatic ring;

wherein in the structure (3):

R^1aselected from:

-C_1-4alkyl optionally substituted with one or more groups independently selected from: OH; c_1-4Alkoxy optionally substituted by one or more independently selected OH or C_1-4Alkoxy substitution; -SO₂-C_1-4An alkyl group; -O-C_3-7A monocyclic cycloalkyl group; and-O-heterocycloalkyl, wherein the heterocycloalkyl is a 4 to 7 membered monocyclic heterocycloalkyl containing one, two or three heteroatoms independently selected from N, O and S;

-NR^6aR^6b；

-C_1-4an alkoxy group;

-C_3-7a monocyclic cycloalkyl group;

a 4 to 7 membered monocyclic heterocycloalkyl comprising one, two or three heteroatoms independently selected from N, O and S, optionally substituted with one or more halo;

-O-C_3-7monocyclic cycloalkyl, and

-O-heterocycloalkyl, wherein the heterocycloalkyl is a 4 to 7 membered monocyclic heterocycloalkyl comprising one, two or three heteroatoms independently selected from N, O and S;

R^1bis H or C_1-4An alkyl group;

Cy₁is a 5 membered monocyclic heteroaryl ring comprising one, two or three heteroatoms independently selected from N, O or S, or

Cy₁Is a 4 to 7 membered monocyclic heterocycloalkyl ring comprising one, two or three heteroatoms independently selected from N, O or S, or a 4 to 7 membered monocyclic heterocycloalkyl ring comprising one, two or three heteroatoms independently selected from N, O or S, fused to a 5 to 6 membered heteroaryl ring comprising one, two or three heteroatoms independently selected from N, O or S, which heteroaryl may optionally be substituted with one C_1-4Alkyl substitution;

R³selected from:

-C_1-4alkyl optionally substituted with one or more independently selected groups: halogen; -CN; -OH; -C_1-4An alkoxy group; or-NR^7aR^7b；

-C_1-4Alkoxy substituted with one or more halogens;

-C_3-7a monocyclic cycloalkyl group;

-a 4 to 7 membered monocyclic heterocycloalkyl comprising one, two or three heteroatoms independently selected from N, O and S;

-CN；

-S(O)₂-C_1-4an alkyl group;

-NR^8aR^8b(ii) a And

-C(＝O)NR^8cR^8d；

each R⁴Independently selected from: -C_1-4Alkyl optionally substituted by one or more independently selected R¹²Substituted by groups; -C_3-7A monocyclic cycloalkyl group; and 4 to 7 membered monocyclic heterocycloalkyl comprising one, two or three heteroatoms independently selected from N, O and S;

each R¹²Independently selected from: -a halogen; -OH; -C_1-4An alkoxy group; -SO₂-C_1-4An alkyl group; -C_3-7Monocyclic cycloalkyl optionally substituted by one or more independently selected-OH, halogen, -CN, C_1-4Alkyl radical, C_1-4Alkoxy or ═ O substitution; a 4 to 7 membered monocyclic heterocycloalkyl comprising one, two or three heteroatoms independently selected from N, O and S, optionally substituted with one or more independently selected-OH, halo, -CN, C_1-4Alkyl radical, C_1-4Alkoxy or ═ O substitution; -NR^9aR^9b(ii) a and-CN;

each R^6a、R^6b、R^7a、R^7b、R^8a、R^8b、R^8c、R^8d、R^9a、R^9bIndependently selected from H and C_1-4An alkyl group;

subscript n is 0, 1,2, or 3;

in some embodiments, when Ar²In the case of the above structure (3), X is-CR^bR^cA; y and W are-NR^aA; z is-C (O) -or-C (═ N-CN) -.

In some embodiments, in said formula III:

R¹represents (1) optionally substituted by 1-5R²¹Substituted C_1-8Alkyl, or (2) optionally substituted by 1-5 substituents selected from C_1-4Alkyl radical, C_1-4C substituted by substituent groups of alkoxy, halogen atom and trifluoromethyl_3-7A carbocyclic ring;

in some embodiments, R¹Is C_1-8An alkyl group; in some embodiments, R¹Is C_1-6An alkyl group; in some embodiments, R¹Is (CH)₃)₂CHCH₂-or (CH)₃CH₂)₂CH-；

In some embodiments, R²Is a hydrogen atom;

in some embodiments, R¹⁴Is a hydroxyl group;

in some embodiments, M¹And M²Each independently represents-O-or-S-;

in some embodiments, R³And R⁴Each independently represents (1) a halogen atom, (2) C optionally substituted by halogen_1-4Alkoxy, or (3) -L-SO₂R⁸；

In some embodiments, L represents a bond;

in some embodiments, R⁸Is represented by C_1-4An alkyl group;

in some embodiments, R⁸Is methyl;

in some embodiments, R³And R⁴Each independently represents F or-SO₂CH₃or-OCH₂CH₂F；

In some embodiments, ring 1 and ring 2 each independently represent a 5-7 membered ring group selected from: aryl, cycloalkyl or heteroaryl; optionally, ring 1 and ring 2 each independently represent an aryl group; in some embodiments, ring 1 and ring 2 each independently represent phenyl, cyclohexyl, pyridyl; in some embodiments, ring 1 and ring 2 are each independently phenyl;

in some embodiments, E is trifluoromethyl;

in some embodiments, m and n are both 2.

In some embodiments, in said formula II:

R^a、R^band R^cEach independently selected from: hydrogen; or alkyl optionally substituted with independently selected hydroxyl, optionally substituted alkoxy, optionally substituted ester, optionally substituted aminocarbonyl, or carboxyl;

in some embodiments, R^a、R^bAnd R^cEach independently selected from: hydrogen; or C optionally substituted by a group selected from hydroxy, optionally substituted_1-4C substituted by alkoxy, optionally substituted ester, optionally substituted aminocarbonyl, or carboxyl groups_1-6An alkyl group;

in some embodiments, R^a、R^bAnd R^cEach independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl, or carboxy_1-6An alkyl group;

in some embodiments, R^a、R^bAnd R^cEach independently selected from: hydrogen, methyl, ethyl, -CH₂COOH、-CH₂CH₂OH、-CH₂CH₂CH₂OH、-CH₂CONH₂Or CH₂COOEt；

In some embodiments, R^a、R^bAnd R^cEach independently selected from hydrogen or methyl;

in some embodiments, W is-NR^a—；

In some embodiments, Z is-c (o) -;

in some embodiments, Y is-NR^aA; and

in some embodiments, X isDirect bond, — NR^a-, or-CR^bR^c—；

In some embodiments of the present invention, the substrate is,

is-NH-NH-C (O) -NH-, -N (CH)₃) -NH-C (O) -NH-, -NH-C (O) -NH-or-CH₂-NH-C(O)-NH-；

In some embodiments, at Ar¹Optionally substituted aryl appearing in the definition of (1) is optionally substituted C₆-C₁₀An aryl group;

in some embodiments, at Ar¹Optionally substituted heteroaryl groups appearing in the definition of (a) are optionally substituted monocyclic or fused bicyclic heteroaryl groups;

in some embodiments, at Ar¹Optionally substituted heteroaryl as appearing in the definition of (a) is an optionally substituted monocyclic 5-10 or 5-to 7-membered heteroaryl or an optionally substituted fused bicyclic 8-to 12-membered heteroaryl;

in some embodiments, Ar¹Is optionally substituted phenyl or optionally substituted 5 to 6 membered monocyclic heteroaryl comprising one or two heteroatoms independently selected from N, O and S;

in some embodiments, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; alkyl optionally substituted with halogen, hydroxy or alkoxy; alkoxy optionally substituted with halogen, hydroxy or alkoxy; or amino optionally substituted with alkyl;

in some embodiments, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; optionally substituted by halogen, hydroxy or C_1-6Alkoxy-substituted C_1-6An alkyl group; optionally substituted by halogen, hydroxy or C_1-6Alkoxy-substituted C_1-6An alkoxy group; or optionally is covered with C_1-6An alkyl-substituted amino group;

in some embodiments, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; c optionally substituted by halogen, hydroxy_1-6An alkyl group; optionally substituted by hydroxy, C_1-6Alkoxy-substituted C_1-6An alkoxy group; or optionally is covered with C_1-6An alkyl-substituted amino group;

in some embodiments, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; c optionally substituted by halogen_1-6An alkyl group; optionally substituted by hydroxy, C_1-6Alkoxy-substituted C_1-6An alkoxy group;

in some embodiments, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen, fluorine, chlorine, bromine, trifluoromethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH₂CH₂OCH₃，-OCH₂CH(OH)CH₂OH, N-ethyl-amino or N-ethyl-N-methyl-amino;

in some embodiments, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen, chlorine, bromine, trifluoromethyl, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH₂CH(OH)CH₂OH or-OCH₂CH₂OCH₃；

In some embodiments, at Ar¹Said heteroaryl group as appearing in the definition of (1) is optionally substituted with 1 to 2 of said substituents in ortho position to the heteroatom;

in some embodiments, at Ar¹The heteroaryl group appearing in the definition of (1) is pyridine or thiophene;

in some embodiments, Ar¹Selected from the group consisting of 4-substituted 2, 6-dichloropyridine, 4-substituted 2, 6-dibromopyridine, 4-substituted 2-chloro-6-hydroxyethylpyridine, 4-substituted-2-chloro-6-hydroxypropylpyridine, 4-substituted 2-chloro-6-methoxypyridine, 4-substituted 2-chloro-6-ethoxypyridine, 4-substituted-2-chloro-6-n-propoxypyridine, 4-substituted-2-chloro-6-isopropoxypyridine, 4-substituted-2-chloro-6-n-butoxypyridine, 4-substituted-2-chloro-6- (2-methoxyethoxy) pyridine, and mixtures thereof, 4-substituted-2-trifluoromethyl-6- (2-methoxyethoxy) pyridine, 4-substituted-2-trifluoromethyl-6-ethoxypyridine, 4-substituted-2-trifluoromethyl-6-(2, 3-dihydroxypropoxy) pyridine, 4-substituted-2- (N-ethylamino) -6-chloropyridine, 4-substituted-2- (N-ethyl-N-methylamino) -6-chloropyridine, or 5-substituted-2, 3-dichlorothiophene;

in some embodiments, Ar¹Is 4-substituted 2, 6-dichloropyridine, 4-substituted-2-chloro-6- (2-methoxyethoxy) pyridine, 4-substituted-2-trifluoromethyl-6-ethoxypyridine or 4-substituted-2-trifluoromethyl-6- (2, 3-dihydroxypropoxy) pyridine;

in some embodiments, Ar¹And Ar²Are each 4-substituted 2, 6-dichloropyridines;

in some embodiments, Ar¹Is aryl optionally substituted with s groups independently selected from: hydrogen, halogen, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, optionally substituted cycloalkoxy or optionally substituted cycloalkyl;

in some embodiments, Ar¹Is aryl optionally substituted with s groups independently selected from: hydrogen, halogen, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Haloalkyl, C_1-6Haloalkoxy, amino, mono-or di-C_1-6Alkylamino radical, C_3-7Cycloalkyl or C_3-7A cycloalkoxy group.

In some embodiments, Ar¹Is aryl optionally substituted with s groups independently selected from: hydrogen, halogen, C_1-6Alkyl radical, C_1-4Alkoxy radical, C_1-6Perhaloalkyl, C_1-4Perhaloalkoxy, amino, mono-or di-C_1-6Alkylamino radical, C_3-7Cycloalkyl or C_3-7A cycloalkoxy group.

In the structure (1) of the formula II:

in some embodiments, R₁Is optionally substituted C_1-12An alkyl group; in some embodiments, R₁Selected from C optionally substituted by halogen, alkenyl or hydroxy_1-6An alkyl group; in some embodiments, R₁Selected from C optionally substituted by alkenyl or hydroxy_1-6An alkyl group; in some embodiments, R₁Is methyl, ethyl, n-propyl, isopropyl, allyl or-CH₂CH₂CH₂OH; in some embodiments, R₁Is methyl, allyl or-CH₂CH₂CH₂OH; in some embodiments, R₁Is methyl;

in some embodiments, R₂Selected from hydrogen, deuterium, halogen, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Perhaloalkyl radical, C_1-6Perhaloalkoxy, amino, mono-or di-C_1-6Alkylamino radical, C_3-7Cycloalkyl or C_3-7A cycloalkoxy group; in some embodiments, R₂Selected from hydrogen and C_1-6An alkyl group; in some embodiments, R₂Selected from hydrogen and methyl;

in some embodiments, R₃And R₄Each independently selected from hydrogen, deuterium, halogen, -CN, -NO₂Nitroso group, carboxyl group, sulfonic acid group, optionally substituted sulfonyl group, optionally substituted C_1-6Alkyl, optionally substituted C_1-6Alkoxy, optionally substituted amino, optionally substituted C_3-7Cycloalkyl, optionally substituted C_3-7Cycloalkoxy, optionally substituted C₆-C₁₈Aryl or optionally substituted C₁-C₁₈A heteroaryl group;

in some embodiments, R₃And R₄Each independently selected from: hydrogen; and C independently of one another optionally substituted by_1-6Alkyl radical, C_3-7Cycloalkyl radical, C₆-C₁₈Aryl or C₁-C₁₈Heteroaryl group: halogen, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl or C_1-6An alkoxy group;

in some embodiments, R₃And R₄Each independently selected from: hydrogen; c_3-7A cycloalkyl group; optionally substituted by halogen, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl or C_1-6Alkoxy-substituted C_1-6An alkyl group; each independently of the others optionally substituted by halogenOr C_1-6Alkoxy-substituted C₆-C₁₈Aryl or C₁-C₁₈A heteroaryl group;

in some embodiments, R₃And R₄Each independently selected from: hydrogen; c_3-7A cycloalkyl group; optionally substituted by halogen or C₆-C₁₈Aryl substituted C_1-6An alkyl group; optionally substituted by halogen, or C_1-6Alkoxy-substituted C₆-C₁₈An aryl group; or C optionally substituted by halogen₁-C₁₈A heteroaryl group;

in some embodiments, R₃And R₄Each independently selected from: hydrogen; c_3-6A cycloalkyl group; c optionally substituted by halogen or phenyl_1-6An alkyl group; optionally substituted by halogen, or C_1-6Alkoxy-substituted phenyl; or furyl or thienyl each independently optionally substituted with halogen;

in some embodiments, R₃And R₄Each independently selected from: hydrogen, methyl, n-propyl, isopropyl, cyclopropyl, cyclohexyl, -CF₃Benzyl, phenyl, fluoro-substituted phenyl, methoxy-substituted phenyl, furyl or chloro-substituted thienyl;

in some embodiments, R₃And R₄Is not hydrogen and is located at h.

Wherein, in the structure (2) of the formula II:

in some embodiments, the structure (2) is represented by the following structure (2-1):

in the structure (2-1), R¹And R²Independently selected from H and alkyl, methoxy, hydroxy, halogen, nitrile, trifluoromethyl and carboxy;

R³independently selected from alkyl, methoxy, hydroxy, halogen, nitrile, trifluoromethyl and carboxy;

d is CR or N;

x is O, NR⁴，CR⁴R⁵Wherein R is present in the definition of X⁴And R⁵Independently selected from: h and alkyl groups such as lower alkyl groups and branched alkyl groups which may have 1 to 10 carbons and may be cyclic or branched alkyl groups having 3 to 10 carbons; a methoxy group; a hydroxyl group; f; br; i; a nitrile; trifluoromethyl and carboxyl;

y is O or S;

z is a substituted aromatic ring having the structure:

wherein R is₆And R₇Independently selected from: alkyl groups and may include 1 to 10 carbons and may be cyclic or branched alkyl groups having 3 to 10 carbons; a methoxy group; a hydroxyl group; halogen; a nitrile; trifluoromethyl and carboxyl; and

e is N or CR;

in some embodiments, the structure (2) is represented by the following structure (2-2):

in the structure (2-2), R¹And R²Independently selected from H and alkyl, methoxy, hydroxy, halogen, nitrile, trifluoromethyl and carboxy;

x is O, NR⁴，CR⁴R⁵Wherein R is⁴And R⁵Independently selected from: h and alkyl groups such as lower alkyl groups and branched alkyl groups which may have 1 to 10 carbons and may be cyclic or branched alkyl groups having 3 to 10 carbons; a methoxy group; a hydroxyl group; f; br; i; a nitrile; trifluoromethyl and carboxyl;

y is O or S;

z is a substituted aromatic ring having the structure:

e is N or CR.

Wherein, in the structure (3) of the formula II:

in some embodiments, R^1aIs selected from methyl;

in some embodiments, R^1bSelected from hydrogen;

in some embodiments of the present invention, the substrate is,

is composed of

In some embodiments, R³And R⁴Each independently selected from hydrogen or C optionally substituted by halogen_1-6An alkyl group;

in some embodiments, R³Is methyl;

in some embodiments, R⁴Is hydrogen or trifluoromethyl; in some embodiments, R⁴At least one of which is trifluoromethyl.

In some embodiments, the S1PR2 inhibitor is the following compound or a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition thereof, or a combination thereof with one or more other therapeutic agents:

In some embodiments, the cell is a cell line, or a cell from a subject;

in some embodiments, the cell is selected from a tumor cell; optionally, the tumor cell is selected from breast cancer cell, liver cancer cell, lung cancer cell, esophageal cancer cell, stomach cancer cell, pancreatic cancer cell, bile duct cancer cell, head and neck tumor cell, nasopharyngeal cancer cell, etc.

In some embodiments, the cell is a cell line, or a cell from a subject;

In a ninth aspect, the present application provides a method of screening for a drug-resistant oncology therapeutic drug, wherein the drug-resistant oncology therapeutic drug is one that is capable of resisting and/or reversing the drug resistance of a fluorouracil-based drug, the method comprising:

providing a cell line or cell culture expressing S1PR 2;

contacting a candidate with the cell line or cell culture;

detecting the expression level of S1PR2 in the cell line or cell culture after contact with the candidate and comparing to the expression level of S1PR2 in a control cancer cell line or cancer cell culture that has not been contacted with the candidate;

when the expression level of S1PR2 is lower than that of the control cancer cell line or cancer cell culture, it indicates that the candidate can be used as a drug-resistant tumor therapeutic drug.

In a tenth aspect, the present application provides a method of screening for a drug-resistant oncology therapeutic drug, wherein the drug-resistant oncology therapeutic drug is one capable of resisting and/or reversing the drug resistance of a fluorouracil-based drug, comprising:

providing a cancer cell line or a cancer cell culture expressing DPD;

contacting a candidate with the cancer cell line or cancer cell culture;

detecting the expression level of DPD in said cell line or cell culture after contact with the candidate and comparing the expression level of DPD with the expression level of DPD of a control cancer cell line or cancer cell culture not contacted with the candidate;

when the expression level of DPD is lower than that of a control cancer cell line or cancer cell culture, indicating that the candidate is capable of acting as a drug-resistant oncology therapy drug;

wherein the candidate is selected from S1PR2 inhibitors.

In an eleventh aspect, the present application provides a kit comprising:

optionally an antineoplastic agent.

In some embodiments, the kit further comprises instructions for use.

Detailed Description

The following examples are given to further illustrate the embodiments of the present invention. The following examples are for illustrative purposes only and are not intended to limit the scope of the present application.

Preparation example 1

The following known inhibitors of S1PR2 were prepared by synthetic methods or synthetic routes reported in literature or patents (including but not limited to those mentioned in the background section above):

preparation example 2:

compounds 9-1 to 9-9 of general formula I of the present invention were synthesized by general synthetic route I of the present invention:

scheme I comprises the following steps:

step 1 a:

step 1:

step 2:

and the following step 3-1 or step 3-2:

step 3-1:

step 3-2:

synthesis of Compound 9-1:

the preparation method is characterized in that 1 methyl-5-aminopyrazole (1) and 2, 6-dichloropyridine-4 formic acid are used as initial raw materials, and the preparation method comprises the following steps: namely 1 methyl-5-aminopyrazole (1,30.93mmol) and ethyl isobutyrylacetate (37.11mmol) take acetic acid as a solvent, reflux is carried out for 20h at 140 ℃, the temperature is cooled to room temperature, the solvent is dried in a spinning mode, 10mL of ethyl acetate is added, stirring is carried out for ten minutes, and then suction filtration is carried out to obtain a white solid 1.

Compound 2(1.95mmol) with POBr₃(2.54mmol) anisole as solvent, refluxing at 120 deg.C for 3h, cooling to room temperature, washing with saturated aqueous sodium chloride solution, extracting with dichloromethane, combining organic layers, and extracting with anhydrous MgSO₄Drying, spin-drying the solvent, and performing column chromatography to obtain a white solid 3.

Compound 3(200mg,0.89mmol), NIS (200mg,0.89mmol), 48% HBF₄(391mg,4.44mmol) in CH₃CN (5mL), reflux at 80 ℃ for 2 h. TLC detection shows that the reaction is complete, and the reaction solution is poured into 1M NaHCO₃In aqueous solution, filtered and washed with water to give compound 4(237mg, 76% yield).¹H NMR(600MHz,DMSO-d₆)δ7.13(s,1H),3.90(s,3H),2.62(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ149.51,145.89,141.64,122.09,116.71,89.24,34.61,17.33.

Compound 4(1.0g,2.85mmol), Cs₂CO₃(1.9g,5.7mmol),PdCl₂(PPh₃)₂(100mg,0.14mmol), and 4-pyridineboronic acid (350mg,2.85mmol) in 1, 4-dioxane/H₂O (15mL, v/v 3/1). The reaction system is deaerated, nitrogen is introduced for protection, and stirring is carried out for 12h at the temperature of 80 ℃. TLC detection shows that the reaction is complete, the reaction solution is washed by water and saturated NaCl aqueous solution, extracted by dichloromethane and anhydrous MgSO₄Drying, filtering, spin-drying the solvent, and performing column chromatography to obtain compound 5-1(410mg, 52%).¹H NMR(500MHz,DMSO-d₆)δ8.81(d,J＝2.3Hz,1H),8.66(d,J＝4.8Hz,1H),8.05(dt,J＝8.0,1.9Hz,1H),7.53(dd,J＝7.9,4.8Hz,1H),7.29(s,1H),4.05(s,3H),2.37(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ150.59,150.20,149.94,146.27,141.52,141.37,137.49,129.60,123.81,122.10,113.11,34.56,19.68.

Refluxing compound 5-1(1.80mmol) and 80% hydrazine hydrate (89.89mmol) in ethanol at 100 deg.C for 10 hr, cooling to room temperature, spin-drying the solvent, washing with saturated aqueous sodium chloride solution, extracting with dichloromethane, mixing organic layers, and extracting with anhydrous MgSO₄Drying and spin-drying the solvent to obtain a white solid 6-1.

Placing 2, 6-dichloropyridine-4-formic acid (26mmol) in a double-neck bottle, adding 30mL1, 4-dioxane under the protection of nitrogen, dropwise adding triethylamine (39mmol), dropwise adding diphenylphosphorylazide DPPA (39mmol) under the condition of ice bath, reacting overnight, detecting the reaction completion by TLC, washing with saturated aqueous sodium chloride solution, extracting with dichloromethane, combining organic layers, and using anhydrous MgSO₄Drying, spin-drying the solvent, and performing column chromatography to obtain a white solid 8.

Dissolving compound 8(1.68mmol) in 3mL of toluene, heating at 80 deg.C for 3h, detecting by TLC to convert into isocyanate, cooling to 50 deg.C, adding tetrahydrofuran solution of compound 6-1(1.68mmol), reacting for 5h, detecting by TLC to complete reaction, cooling to room temperature, spin-drying solvent, washing with saturated sodium chloride solution, extracting with dichloromethane, combining organic layers, and purifying with anhydrous MgSO₄Drying, spin-drying solvent, and performing column chromatography to obtain white solid 9-1.

9-1：¹H NMR(500MHz,DMSO-d₆)δ9.80(d,J＝86.4Hz,1H),9.16–8.74(m,3H),8.63(d,J＝4.8Hz,1H),8.02(dd,J＝7.8,2.1Hz,1H),7.98–7.37(m,3H),6.42(s,1H),3.91(s,3H),2.29(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ159.87,151.97,151.15,149.90,149.85,149.25,143.46,140.88,137.42,130.73,123.76,111.65,108.72,105.13,33.84,20.32.

Other compounds of formula I may be prepared according to the above synthetic routes.

9-2：¹H NMR(500MHz,DMSO-d₆)δ9.70(s,1H),8.93(d,J＝43.5Hz,2H),7.85(dd,J＝60.5,8.1Hz,6H),6.42(s,1H),3.89(s,3H),2.30(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ151.98,149.84,142.38,139.44,132.54,130.67,119.25,110.97,33.90,20.65.

9-3：¹H NMR(500MHz,DMSO-d₆)δ9.77(d,J＝101.5Hz,1H),8.88(d,J＝64.7Hz,2H),8.02–7.02(m,5H),6.33(s,1H),3.86(s,3H),2.04(d,J＝53.6Hz,6H).¹³C NMR(125MHz,DMSO-d₆)δ163.35,161.41,151.97,150.56,149.84,142.23,140.48,132.78,130.89,116.78,112.67,33.67,20.10,18.48.

9-4：¹H NMR(500MHz,DMSO-d₆)δ9.70(s,1H),8.78(s,2H),8.00–7.33(m,4H),7.05–6.88(m,2H),6.35(s,1H),4.06–3.79(m,5H),2.27(s,3H),1.73(m,J＝7.1Hz,2H),0.98(t,J＝7.4Hz,3H).¹³C NMR(125MHz,DMSO-d₆)δ158.98,151.99,150.98,149.83,143.92,131.12,126.89,114.40,69.39,33.63,22.51,20.32,10.8.

9-5：¹H NMR(500MHz,DMSO-d₆)δ9.58(s,2H),8.76(s,2H),7.35(d,J＝8.2Hz,4H),6.83(d,J＝8.2Hz,2H),6.34(s,1H),3.85(s,3H),2.27(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ159.58,157.76,151.97,150.94,149.83,144.28,143.82,131.13,125.38,115.30,111.73,108.58,104.42,33.59,20.29.

9-6：¹H NMR(500MHz,DMSO-d₆)δ9.67(s,1H),8.84(d,J＝94.9Hz,2H),7.96–7.41(m,2H),6.34(s,1H),5.99(d,J＝3.3Hz,1H),4.21(s,2H),3.80(q,J＝4.2,3.1Hz,5H),2.45(s,5H).¹³C NMR(125MHz,DMSO-d₆)δ159.51,151.97,150.83,149.82,144.29,143.84,129.25,128.29,111.71,104.75,65.16,64.13,33.60,28.91,20.56.

9-7：¹H NMR(500MHz,DMSO-d₆)δ9.70(s,1H),8.83(s,2H),8.31(d,J＝2.5Hz,1H),7.86(dd,J＝8.5,2.5Hz,3H),6.81(d,J＝8.5Hz,1H),6.37(s,1H),5.28(m,J＝6.2Hz,1H),3.87(s,3H),2.28(s,3H),1.30(d,J＝6.2Hz,6H).¹³C NMR(125MHz,DMSO-d₆)δ162.99,151.98,151.05,149.83,147.37,141.00,140.53,123.68,110.92,104.82,68.13,33.73,22.34,20.23.

9-8：¹H NMR(500MHz,DMSO-d₆)δ9.69(s,1H),8.80(s,2H),8.02–7.32(m,4H),6.80(d,J＝2.0Hz,1H),6.36(s,1H),3.84(s,3H),2.44(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ159.65,151.99,150.98,149.83,143.70,141.82,135.94,119.66,112.11,111.85,104.64,33.64,20.44.

9-9：¹H NMR(500MHz,DMSO-d₆)δ9.71(s,1H),8.82(s,2H),7.93–7.50(m,4H),7.45–7.36(m,2H),7.21–7.12(m,1H),7.12–7.02(m,4H),6.38(s,1H),3.87(s,3H),2.30(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ157.20,156.71,151.97,151.03,149.84,143.47,131.57,130.56,129.76,124.21,119.49,118.27,33.69,20.42.

Preparation example 3:

compounds of the structure shown below were synthesized:

16-1：¹H NMR(500MHz,DMSO-d₆)δ9.46(d,J＝471.6Hz,1H),8.58(s,2H),7.90(s,1H),7.58(s,2H),7.30–7.09(m,5H),4.09(s,2H),3.80(s,3H),2.36(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ156.96,149.91,148.30,140.93,139.95,131.28,128.68,128.50,126.30,111.27,33.46,30.90,15.90.

16-2：¹H NMR(500MHz,DMSO-d₆)δ9.73(s,1H),8.97(s,2H),7.99(s,1H),7.91–7.44(m,4H),7.38(t,J＝8.8Hz,2H),6.66(s,1H),3.88(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ164.17,162.21,159.94,151.97,150.77,149.83,143.28,133.82,131.57,130.50,130.43,116.78,116.61,111.73,33.89.

16-3：¹H NMR(500MHz,DMSO-d₆)δ9.81(d,J＝88.3Hz,1H),8.95(d,J＝42.2Hz,2H),7.71(d,J＝8.2Hz,5H),7.11(d,J＝8.3Hz,2H),6.63(s,1H),3.84(d,J＝28.7Hz,6H).¹³C NMR(125MHz,DMSO-d₆)δ160.70,159.95,151.99,150.85,149.83,144.07,131.72,105.64,98.04,129.63,115.17,111.82,101.12,55.76,33.86.

16-4：¹H NMR(500MHz,DMSO-d₆)δ9.77(d,J＝88.2Hz,1H),8.77(s,2H),7.87(d,J＝34.2Hz,3H),6.38(s,1H),3.83(s,3H),2.74(t,J＝7.5Hz,2H),1.66(h,J＝7.4Hz,2H),0.89(t,J＝7.3Hz,3H).¹³C NMR(125MHz,DMSO-d₆)δ159.80,151.98,150.15,149.84,147.38,131.18,111.75,110.67,102.85,34.82,33.71,22.99,14.17.

16-5：¹H NMR(400MHz,DMSO-d₆)δ9.97(s,2H),7.55(s,4H).¹³C NMR(100MHz,DMSO-d₆)δ151.63,150.84,150.12,111.93.

16-6：¹H NMR(500MHz,DMSO-d₆)δ9.80(s,1H),8.99(s,2H),8.07–7.34(m,8H),6.74(s,1H),3.91(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ159.98,151.92,150.81,149.87,144.49,137.46,131.62,129.74,129.65,128.26,111.72,108.39,102.03,33.88.

16-7:¹H NMR(500MHz,DMSO-d₆)δ9.76(d,J＝103.0Hz,1H),8.78(s,2H),7.89(d,J＝58.4Hz,3H),6.40(s,1H),3.82(s,3H),3.28–3.08(m,1H),1.28(d,J＝6.9Hz,6H).¹³C NMR(125MHz,DMSO-d₆)δ159.85,153.42,152.01,150.34,149.83,131.22,111.70,100.38,33.70,32.03,22.86.

16-8:¹H NMR(500MHz,DMSO-d₆)δ9.65(s,1H),8.67(s,2H),7.86(d,J＝39.3Hz,3H),6.17(s,1H),3.81(s,3H),2.20(tt,J＝8.7,4.9Hz,1H),1.17–1.05(m,2H),0.92(dt,J＝6.9,3.4Hz,2H).¹³C NMR(125MHz,DMSO-d₆)δ159.80,151.99,150.13,149.82,130.90,111.69,109.59,98.47,33.70,14.18,10.51.

16-9:¹H NMR(500MHz,DMSO-d₆)δ9.69(s,1H),8.75(s,2H),7.87(d,J＝35.1Hz,2H),6.37(s,1H),3.83(s,3H),2.45(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ159.82,152.00,150.00,149.83,131.29,111.75,103.65,55.35,33.74,18.62.

16-10:¹H NMR(500MHz,DMSO-d₆)δ9.73(s,1H),8.93(d,J＝37.5Hz,2H),7.99–7.35(m,7H),6.37(s,1H),3.80(s,3H),2.01(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ159.32,151.97,151.27,149.83,146.74,139.60,138.03,129.09,128.98,128.84,111.70,33.37,15.28.

16-11:¹H NMR(500MHz,DMSO-d₆)δ9.76(d,J＝91.1Hz,1H),8.67(s,2H),7.69(d,J＝136.7Hz,2H),6.26(s,1H),3.72(s,3H),2.49(s,3H),2.45(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ159.74,151.99,150.98,149.81,143.99,140.16,111.69,103.26,33.21,19.23,14.91.

16-12:¹H NMR(500MHz,DMSO-d₆)δ9.72(s,1H),8.89(d,J＝53.2Hz,2H),7.82(s,2H),7.40(d,J＝8.6Hz,2H),7.06(d,J＝8.7Hz,2H),6.33(s,1H),3.79(d,J＝8.8Hz,6H),2.07(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ160.07,159.34,151.98,151.36,149.82,146.56,139.65,130.38,130.20,114.28,111.70,103.19,55.65,33.36,15.50.

16-13:¹H NMR(500MHz,DMSO-d₆)δ9.63(s,1H),8.73(d,J＝106.9Hz,2H),7.69(d,J＝121.4Hz,2H),6.00(s,1H),3.72(s,3H),2.53(s,3H),2.33(td,J＝8.5,8.0,4.0Hz,1H),1.07(h,J＝4.2Hz,2H),0.76–0.68(m,2H).

16-14:¹H NMR(600MHz,DMSO-d₆)δ9.74(d,J＝115.8Hz,1H),8.66(s,2H),7.66(d,J＝162.4Hz,2H),6.26(s,1H),3.70(s,3H),2.76(t,J＝7.8Hz,2H),2.41(s,3H),1.58(h,J＝7.5Hz,2H),0.92(t,J＝7.3Hz,3H).Carbon13-NMR(150MHz,DMSO-D6)δ159.71,152.08,151.30,149.93,139.64,34.47,33.29,23.85,15.08,14.20.

16-15:¹H NMR(600MHz,DMSO-d₆)δ9.77(d,J＝94.0Hz,1H),8.92(d,J＝47.9Hz,2H),7.49(s,4H),7.32(s,2H),6.33(s,1H),3.77(s,3H),2.00(s,3H).¹³C NMR(125MHz,DMSO-d₆)δ163.77,161.82,159.31,151.96,151.27,149.83,145.62,139.54,134.36,131.19,131.13,115.91,115.74,33.38,15.30.

16-16:¹H NMR(400MHz,DMSO-d₆)δ9.69(s,1H),8.71(s,2H),7.72(d,J＝98.5Hz,2H),6.36(s,1H),3.74(s,3H),3.04(s,1H),2.48(s,3H),1.97–1.67(m,5H),1.53–1.33(m,4H),1.33–1.23(m,1H).¹³C NMR(100MHz,DMSO-d₆)δ159.82,153.48,152.06,151.20,149.88,139.22,111.76,99.28,33.55,33.22,26.58,26.15,15.19.

16-17:¹H NMR(400MHz,DMSO-d₆)δ9.76(s,1H),8.99(s,2H),7.81(s,2H),7.44–7.19(m,2H),6.46(s,1H),3.81(s,3H),2.27(s,3H).¹³C NMR(100MHz,DMSO-d₆)δ151.95,151.54,149.88,139.52,137.63,137.50,129.89,129.09,128.26,111.69,33.44,15.67.

16-18:¹H NMR(400MHz,DMSO-d₆)δ9.76(s,1H),8.94(d,J＝38.2Hz,2H),7.96(d,J＝1.7Hz,3H),7.08(d,J＝3.4Hz,1H),6.77–6.63(m,2H),3.81(s,3H),2.48(s,3H).¹³C NMR(100MHz,DMSO-d₆)δ159.39,152.00,151.97,150.77,149.87,145.13,139.69,134.21,112.80,111.85,104.87,99.85,33.39,16.44.

16-19:¹H NMR(400MHz,DMSO-d₆)δ9.51(s,1H),8.82(d,J＝42.5Hz,2H),8.01(s,1H),7.72(d,J＝8.5Hz,2H),7.13(d,J＝8.5Hz,4H),6.65(s,1H),3.93–3.74(m,9H).¹³C NMR(100MHz,DMSO-d₆)δ164.52,160.72,151.73,150.92,144.03,131.74,129.73,129.64,115.20,55.79,54.21,33.88.

16-20:¹H NMR(400MHz,DMSO-d₆)δ9.47(s,1H),8.82(d,J＝48.6Hz,2H),8.01(s,1H),7.73(d,J＝8.5Hz,2H),7.14(d,J＝8.4Hz,4H),6.65(s,1H),5.09(p,J＝6.2Hz,1H),3.87(d,J＝23.7Hz,6H),1.25(d,J＝6.2Hz,6H).¹³C NMR(100MHz,DMSO-d₆)δ163.69,160.71,151.72,150.92,144.02,131.73,129.73,129.63,115.20,68.85,55.78,33.88,22.22.

16-21:¹H NMR(400MHz,DMSO-d₆)δ9.48(s,1H),8.81(d,J＝43.8Hz,2H),8.01(s,1H),7.72(d,J＝8.4Hz,2H),7.46–6.60(m,5H),4.10(t,J＝6.6Hz,2H),3.87(d,J＝21.4Hz,6H),1.68(q,J＝7.1Hz,2H),0.93(t,J＝7.4Hz,3H).¹³C NMR(100MHz,DMSO-d₆)δ164.32,160.72,151.72,150.93,131.74,129.64,115.21,68.08,55.79,33.88,22.21,10.80.

16-22:¹H NMR(400MHz,DMSO-d₆)δ9.49(s,1H),8.83(d,J＝43.4Hz,2H),8.02(s,1H),7.79–7.68(m,2H),7.14(d,J＝8.8Hz,4H),6.65(s,1H),4.15(t,J＝6.6Hz,2H),3.87(d,J＝23.9Hz,6H),1.65(dd,J＝8.4,6.2Hz,2H),1.47–1.31(m,2H),0.91(t,J＝7.4Hz,3H).¹³C NMR(100MHz,DMSO-d₆)δ164.31,160.72,160.17,151.72,150.92,147.99,144.02,131.74,129.73,129.64,115.21,68.07,55.80,33.88,22.21,10.80.

16-23:¹H NMR(400MHz,DMSO-d₆)δ9.52(s,1H),8.86(d,J＝62.1Hz,2H),8.01(s,1H),7.83–7.72(m,2H),7.63–7.48(m,3H),7.44–6.92(m,2H),6.70(s,1H),3.91(s,3H),3.79(s,3H).¹³C NMR(100MHz,DMSO-d₆)δ164.53,160.15,151.72,150.88,148.05,144.40,137.51,131.66,129.86,129.79,128.29,54.21,33.91.

16-24:¹H NMR(400MHz,DMSO-d₆)δ9.70(s,1H),8.97(d,J＝34.2Hz,2H),8.18–7.62(m,5H),7.13(d,J＝8.4Hz,2H),6.66(s,1H),3.87(d,J＝24.9Hz,6H).¹³C NMR(100MHz,DMSO-d₆)δ160.73,160.00,151.30,150.89,144.10,140.39,131.76,129.66,115.19,55.78,33.89.

example 1: in vitro cell activity assay for S1PR2 inhibitor

The known compounds listed in preparation example 1 above and the compounds synthesized in preparation examples 2 and 3 of the present invention were subjected to in vitro cell activity experiments.

In vitro cell activity assay: DPD expression level verification by prophase pharmacodynamic activity screening and Western Blotting (Western Blotting) in various colon cancer and normal colon cell lines confirms that HCT116 and NCM460 are cell lines with remarkable 5-FU resistance. The log phase growth of HCT116 and NCM460 cells were collected, cell suspension concentration was adjusted, 100. mu.L was added to each well, and the test cells were plated to 1000-10000 wells, (the marginal wells were filled with sterile PBS). 5% CO₂Incubate at 37 ℃ until the cell monolayer is confluent at the bottom of the well (96-well flat bottom plate), add a concentration gradient of S1PR2 inhibitor compound and a constant concentration of 5-FU. Set up 6 concentration gradients, 100 μ L per well, for 3 replicates. 5% CO₂Incubated at 37 ℃ for 24h and observed under an inverted microscope. mu.L of MTT solution (5mg/mL, i.e., 0.5% MTT) was added to each well and incubation was continued for 4 h. The culture was terminated and the culture medium in the wells was carefully aspirated. Add 150 μ L of dimethyl sulfoxide into each well, and shake on a shaker at low speed for 0.5min to dissolve the crystals completely. The absorbance of each well was measured at OD 560nm of an ELISA. Cell viability was expressed as T/C%, T being the OD of the dosed cells and C being the OD of the control cells. Calculation of EC from cell viability at each concentration gradient₅₀And IC₅₀The results are shown in Table 1.

TABLE 1

Note: EC (EC)₅₀Is EC of S1PR2 inhibitor in combination with 5-FU₅₀Values wherein the 5-FU concentration is 20. mu.M. IC (integrated circuit)₅₀For IC of compounds alone₅₀Values for 5-FU without S1PR2 inhibitor, DMSO with solvent, HCT116 and NCM460 with 5-FU resistant cells- -indicate no cell death or no detection.

TABLE 2

Note: EC (EC)₅₀Is EC of S1PR2 inhibitor in combination with 5-FU₅₀Values wherein the 5-FU concentration is 20. mu.M. IC (integrated circuit)₅₀For IC of compounds alone₅₀Values for 5-FU without S1PR2 inhibitor, DMSO with solvent, SW620 with 5-FU resistant cells- -indicate no cell death or no detection.

The result shows that the phenomenon of 5-FU resistance appears in two cells under the action of 20 mu M5-fluorouracil, the S1PR2 inhibitor and the compound can reverse the drug resistance of 5-fluorouracil, and the compound can effectively inhibit the proliferation of drug-resistant cells when being combined with 5-fluorouracil.

Example 2: experiment for inhibiting DPD expression in vitro by S1PR2 inhibitor

HCT116 cells (solvent control group, 10. mu.M administration group to which the compound of Table 1 was administered alone) and SW620 cells (solvent control group, 10. mu.M administration group to which the compound of Table 2 was administered alone (0.6. mu.M administration group was selected as 16-5 compound)) were each taken out and subjected to cell scraping, protein extraction and quantification.

Extracting cell protein: firstly, washing cells by PBS, precooling each group of cells at 4 ℃, and preparing cell lysate: NP-40 (960. mu.L) + PPI (40. mu.L) + PMSF (40. mu.L). PBS was discarded, and 100-. The cells were scraped off with a cell scraper and collected in a 1.5mL centrifuge tube, 12000rpm, 15min for centrifugation. The supernatant was collected in another 1.5mL centrifuge tube, and 90. mu.L of G250 staining solution was added to a 96-well plate, and 10. mu.L of the protein standard solution and 10. mu.L of the sample were added to each tube to prepare a duplicate. The microplate reader determines the cell concentration, adjusts the cell concentration of each group to be consistent, adds 5 Xloading buffer solution to make the concentration to be 1 Xand boils for 5min in a metal bath.

Western blot detection: and (3) preparing separation gel and concentrated gel, and soaking in an electrophoresis solution. An equal amount of 20. mu.g protein sample was added to the loading wells and labeled with either tag or 1 Xloading buffer. After the protein fell down in the separating gel, the constant voltage electrophoresis at 80V was changed to 120V and the constant voltage electrophoresis was continued. The PVDF membrane was used for 100V, 90min membrane rotation. The PVDF membrane was washed once in 5min for 2 times in 1 XTSST on a shaker. The PVDF membrane is sealed by a sealing solution for 1h, and DPD primary antibody is incubated at 4 ℃ overnight. The PVDF membrane was washed with 1 XTSSt on a shaker for 3 times, once for 10 min. The secondary antibody was incubated for 40 min. The PVDF membrane was washed with 1 XTSSt on a shaker for 3 times, once for 10 min. ECL luminescence solution was prepared, and uniformly sprayed on PVDF membrane, and exposed by ChemiDoc XRS and molecular imager (molecular imager). Calculating the gray scale ratio of the analysis imprinting of the Bio-Rad Quantity One, and calculating a formula: relative expression level of protein of interest ═ (grey value of protein of interest-background grey value)/(grey value of internal control protein-background grey value).

The results show that S1PR2 inhibitors are effective in reducing DPD expression in this cell compared to the internal control (see figure 1).

Example 3: in vivo reversion 5-fluorouracil drug resistance experiment

5-fluorouracil was administered in combination with an inhibitor of S1PR2 using a nude mouse subcutaneous inoculation tumor model.

6-8 week old BALB/c nude mice were purchased, 6 each per group, and were housed in a specific pathogen free clean (SPF) environment. Inoculating the 5-FU drug-resistant HCT116 cells subcutaneously, after two weeks, increasing the tumor mass to a certain size, performing aseptic separation and uniform segmentation on the tumor mass, inoculating the tumor mass to the armpit of each nude mouse, feeding the nude mice in an aseptic environment for two weeks, and starting drug administration after the tumor masses of all groups are consistent in size. 20mg/kg 5-fluorouracil combined with 25mg/kg JTE-013 and 25mg/kg 16-6 by tail vein injection for three consecutive and half days, compared to the 5-fluorouracil single group and the solvent group, wherein the solvent volume ratio is 2% DMSO + 30% PEG 300+ 5% Tween 80+ ddH₂Nude mice were recorded for weight and tumor volume changes throughout (measured every 4 days). The mice were then sacrificed and tumor samples and other tissues were removed for relevant detection and analysis of protein expression.

The result shows that the nude mouse tumor inoculated with the HCT116 cell strain has obvious drug resistance to 5-fluorouracil treatment, the in vivo inhibition rate of 5-fluorouracil is only 13.01%, and the inhibition rate of the compound 16-6 can be improved to 46.52%. JTE-013 can significantly improve the sensitivity of tumor cells to 5-fluorouracil treatment, and the inhibition rate is increased to 75.87%, so that S1PR2 inhibitors such as JTE-013 and compounds 16-6 can effectively reverse the drug resistance of tumor cells to 5-fluorouracil (see FIG. 2, wherein NC is a solvent group without 5-FU and compound, and only DMSO and water are given).

Example 4: inhibition of DPD expression in vivo

The tumor tissue, normal colon and liver tissue DPD expression difference of each group of nude mice are subjected to Western blot detection.

Respectively taking out tumor tissues, normal colon and liver tissues of each group of nude mice, then carrying out tissue grinding, protein extraction and quantification, carrying out DPD and beta actin (beta-actin) antibody incubation overnight after SDS-PAGE electrophoresis, membrane transfer and sealing, then carrying out membrane washing and secondary antibody incubation, and developing the PVDF membrane after membrane washing again.

The results showed that 5-fluorouracil had no significant effect on DPD expression in each tissue compared to the internal control, while JTE-013 and compound 16-6 were effective in reducing DPD expression levels in tumor tissues, normal colon and liver tissues (see FIG. 3, where NC is the solvent group without 5-FU and compound, only DMSO and water).

In the in vitro experiments, part of S1PR2 inhibitor is taken as an example, in vivo experiments, JTE-013 (compound 14) and compound 16-6 are taken as examples, and the S1PR2 inhibitor is verified to be capable of effectively inhibiting the expression of DPD, reversing the drug resistance of 5-fluorouracil and effectively inhibiting the tumor growth by being combined with the 5-fluorouracil. The invention also verifies that other S1PR2 inhibitors have similar effects by the same method, and the experimental process is not detailed.

Example 5: knocking down S1PR2 inhibiting DPD expression condition in vitro

shS1PR2 lentivirus knockdown is respectively carried out on the colon cancer cell lines HCT116, SW620, the liver cancer cell Huh7 and the normal liver cell cccHEL-1, and the protein imprinting detection is carried out on the DPD expression difference of each cell line.

Construction and packaging of the short hairpin RNA lentivirus of S1PR2 was provided by Beijing Saisakusho, wherein the S1PR2 knockdown sequence was AGGAACAGCAAG TTCTACTCA. Cells were plated at 4X 10 per well 24h prior to infection⁴Inoculating to 6-well plate, culturing in RPMI-1640 culture medium containing 10% FBS, infecting with lentivirus when cell grows to about 50% density after cell adherence, adding filtered virus solution into 6-well plate, and culturing in 5% CO₂Culturing in a 37 ℃ incubator, observing by using a fluorescence microscope after the virus is infected for 48 hours, and screening by using a 10% FBS RPMI-1640 culture medium added with puromycin to obtain a stable knockdown cell line, wherein the infected cells show green fluorescence. Then, western blot detection is carried out, and the expression difference of S1PR2 and DPD of each cell line is obtained.

The results show that compared with the internal control, S1PR2 of the four cell lines HCT116, SW620, Huh7 and cccHEL-1 are significantly reduced compared with the empty vector group, and have higher knockdown efficiency. Meanwhile, the expression level of DPD of the four cell lines is also obviously reduced (see FIG. 4), which shows that the knockdown of S1PR2 can effectively reduce the expression of DPD, and further proves that an inhibitor of S1PR2 can effectively reduce the expression of DPD.

In conclusion, the invention discovers that the DPD expression level of cancer cells can be regulated or inhibited through targeted inhibition or regulation of the expression level of S1PR2, and the DPD expression level can be prevented from degrading into 5-fluorouracil of the cancer cells, so that the effect of reversing the drug resistance of 5-FU is realized. Since other 5-fluorouracil derivatives can enter blood and finally enter cancer cells through degradation into 5-FU, inhibitors that inhibit DPD production by cancer cells by targeted inhibition or modulation of S1PR2 expression are suitable for drug resistant treatment regimens for cancer treatment with fluorouracil drugs.

The invention has been described in detail with respect to a general description and specific embodiments thereof, but it will be apparent to those skilled in the art that modifications and improvements can be made based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt, or tautomer thereof:

wherein:

R₂selected from hydrogen or optionally substituted alkyl;

ring B is an aromatic or heteroaromatic ring;

x is an integer from 1 to 2; and

y is an integer of 1,2 or 3.

2. The compound of claim 1, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R₁selected from aryl, heteroaryl, or heterocycloalkenyl each independently optionally substituted with halogen, -OH, -CN, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl, alkoxy, aryloxy, or heteroaryloxy;

optionally, optionally，R₁Selected from the group consisting of independently optionally substituted halogen, -OH, -CN, C_1-6Alkyl radical, C_1-6Haloalkyl, C_1-6Hydroxyalkyl, 3-to 10-membered cycloalkyl, C_1-6Alkoxy radical, C₆-C₁₈Aryloxy radical or C₁-C₁₈Heteroaryloxy-substituted C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl, or 5-10 membered heterocycloalkenyl;

optionally, R₁Selected from the group consisting of independently optionally substituted halogen, -OH, -CN, C_1-6Alkyl radical, C_1-6Alkoxy or C₆-C₁₈Aryloxy substituted C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl, or 5-10 membered heterocycloalkenyl;

optionally, R₁Selected from C, each independently optionally substituted with fluoro, chloro, -OH, -CN, methyl, ethyl, n-propyl, isopropyl, n-propoxy, isopropoxy or phenoxy₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl, or 5-10 membered heterocycloalkenyl;

optionally, R₁Selected from C, each independently optionally substituted with fluoro, -OH, -CN, methyl, n-propoxy, isopropoxy or phenoxy₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl, or 5-10 membered heterocycloalkenyl;

optionally, R₁Selected from: optionally selected from halogen, -OH, -CN, C_1-6Alkyl radical, C_1-6Alkoxy, or C₆-C₁₈C substituted by aryloxy radicals₆-C₁₈An aryl group; optionally is covered with C_1-6Alkoxy-substituted C₁-C₁₈A heteroaryl group; or 5-10 membered heterocycloalkenyl;

optionally, R₁Said C occurring in the definition of₆-C₁₈Aryl is phenyl;

optionally, R₁Said C occurring in the definition of₁-C₁₈Heteroaryl is selected from pyridyl or furyl;

optionally, the 5-10 membered heterocycloalkenyl is

Optionally, the halogen is selected from fluorine or chlorine;

optionally, R₁Selected from: phenyl optionally substituted with a group selected from fluoro, -OH, -CN, methyl, n-propoxy, or phenoxy; an isopropoxy substituted pyridyl group; a furyl group; or

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R₂selected from hydrogen, optionally substituted C_1-6An alkyl group;

optionally, R₂Selected from optionally substituted C_1-6An alkyl group;

optionally, R₂Selected from C optionally substituted by halogen, alkenyl or hydroxy_1-6An alkyl group;

optionally, R₂Selected from C optionally substituted by alkenyl or hydroxy_1-6An alkyl group;

optionally, R₂Is methyl, ethyl, n-propyl, isopropyl, allyl or-CH₂CH₂CH₂OH；

Optionally, R₂Is methyl, allyl or-CH₂CH₂CH₂OH；

Optionally, R₂Is methyl.

4. The compound of any one of claims 1 to 3, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

each R₃Independently selected from hydrogen, deuterium, halogen, -CN, -NO₂Nitroso group, carboxyl group, sulfonic acid group, optionally substituted sulfonyl group, optionally substituted C_1-6Alkyl, optionally substituted C_3-8Cycloalkyl, optionally substituted C₆-C₁₈Aryl, heteroaryl, and heteroaryl,Optionally substituted C₁-C₁₈A heteroaryl group;

optionally, each R₃Independently selected from: hydrogen; and each independently optionally halogen, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl or C_1-6Alkoxy-substituted C_1-6Alkyl radical, C_3-8Cycloalkyl radical, C₆-C₁₈Aryl or C₁-C₁₈A heteroaryl group;

optionally, each R₃Independently selected from: hydrogen; c_3-8A cycloalkyl group; optionally substituted by halogen, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl or C_1-6Alkoxy-substituted C_1-6An alkyl group; each independently optionally substituted by halogen, or C_1-6Alkoxy-substituted C₆-C₁₈Aryl or C₁-C₁₈A heteroaryl group;

optionally, each R₃Independently selected from: hydrogen; c_3-8A cycloalkyl group; optionally substituted by halogen or C₆-C₁₈Aryl substituted C_1-6An alkyl group; optionally substituted by halogen, or C_1-6Alkoxy-substituted C₆-C₁₈An aryl group; or C optionally substituted by halogen₁-C₁₈A heteroaryl group;

optionally, each R₃Independently selected from: hydrogen; c_3-8A cycloalkyl group; c optionally substituted by halogen or phenyl_1-6An alkyl group; optionally substituted by halogen, or C_1-6Alkoxy-substituted phenyl; or furyl or thienyl each independently optionally substituted with halogen;

optionally, each R₃Independently selected from hydrogen, methyl, n-propyl, isopropyl, cyclopropyl, cyclohexyl, -CF₃Benzyl, phenyl, fluoro-substituted phenyl, methoxy-substituted phenyl, furyl or chloro-substituted thienyl;

optionally, each R₃Independently selected from hydrogen, or methyl;

optionally, at least one R₃Is not hydrogen and is para to N;

optionally, x is 1.

5. The compound of any one of claims 1 to 4, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R₄、R₅and R₆Each independently selected from: hydrogen; or an alkyl group optionally substituted with a group selected from a hydroxyl group, an optionally substituted ester group, an optionally substituted aminocarbonyl group, or a carboxyl group;

optionally, R₄、R₅And R₆Each independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, optionally substituted ester group, optionally substituted aminocarbonyl, or carboxy_1-6An alkyl group;

optionally, R₄、R₅And R₆Each independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl, or carboxy_1-6An alkyl group;

optionally, R₄、R₅And R₆Each independently selected from: hydrogen, methyl, ethyl, -CH₂COOH、-CH₂CH₂OH、-CH₂CH₂CH₂OH、-CH₂CONH₂Or CH₂COOEt；

Optionally, R₄、R₅And R₆Each independently selected from hydrogen or methyl;

optionally, R₄、R₅And R₆Are all hydrogen.

6. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt, or tautomer thereof, wherein

Optionally, the heteroaromatic ring present in the definition of ring B is C₁-C₁₈A heteroaromatic ring;

optionally, the heteroaryl ring present in the definition of ring B is a 5-10 membered heteroaryl ring;

optionally, the aromatic ring present in the definition of ring B is C₆-C₁₈An aromatic ring;

optionally, determination of Ring BThe aromatic ring in the meaning is C₆-C₁₀An aromatic ring;

optionally, the aromatic rings present in the definition of ring B are benzene rings;

optionally, ring B is a heteroaryl ring;

optionally, the heteroaromatic ring present in the definition of ring B is pyridine or thiophene;

optionally, the heteroaromatic ring present in the definition of ring B is pyridine;

optionally, the heteroaromatic ring present in the definition of ring B is optionally substituted with 1-2R in the ortho position to the heteroatom₇Substituted by groups;

optionally, the step of (a) is carried out,

optionally, the step of (a) is carried out,

selected from the group consisting of 4-substituted 2, 6-dichloropyridine, 4-substituted 2, 6-dibromopyridine, 4-substituted 2-chloro-6-hydroxyethylpyridine, 4-substituted-2-chloro-6-hydroxypropylpyridine, 4-substituted 2-chloro-6-ethoxypyridine, 4-substituted-2-chloro-6-n-propylpyridine, 4-substituted-2-chloro-6-isopropoxypyridine, 4-substituted-2-chloro-6-n-butyloxypyridine, 4-substituted-2-chloro-6- (2-methoxyethoxy) pyridine, 4-substituted-2-trifluoromethyl-6- (2-methoxyethoxy) pyridine, and mixtures thereof, 4-substituted-2-trifluoromethyl-6-ethoxypyridine, 4-substituted-2-trifluoromethyl-6- (2, 3-dihydroxypropoxy) pyridine, 4-substituted-2- (N-ethylamino) -6-chloropyridine, 4-substituted-2- (N-ethyl-N-methylamino) -6-chloropyridine, or 5-substituted-2, 3-dichlorothiophene;

optionally, the step of (a) is carried out,

is 4-substituted 2, 6-dichloropyridine.

7. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt, or tautomer thereof, wherein

Optionally, each R₇Independently selected from hydrogen; halogen; alkyl optionally substituted with halogen, hydroxy or alkoxy; alkoxy optionally substituted with halogen, hydroxy or alkoxy; or amino optionally substituted with alkyl;

optionally, each R₇Independently selected from hydrogen; halogen; optionally substituted by halogen, hydroxy or C_1-6Alkoxy-substituted C_1-6An alkyl group; optionally substituted by halogen, hydroxy or C_1-6Alkoxy-substituted C_1-6An alkoxy group; or optionally is covered with C_1-6An alkyl-substituted amino group;

optionally, each R₇Independently selected from hydrogen; halogen; c optionally substituted by halogen or hydroxy_1-6An alkyl group; optionally substituted by hydroxy or C_1-6Alkoxy-substituted C_1-6An alkoxy group; or optionally is covered with C_1-6An alkyl-substituted amino group;

optionally, each R₇Independently selected from hydrogen; halogen; c optionally substituted by halogen_1-6An alkyl group; optionally substituted by hydroxy or C_1-6Alkoxy-substituted C_1-6An alkoxy group;

optionally, each R₇Independently selected from hydrogen, fluorine, chlorine, bromine, trifluoromethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH₂CH₂OCH₃，-OCH₂CH(OH)CH₂OH, N-ethyl-amino or N-ethyl-N-methyl-amino;

optionally, each R₇Independently selected from hydrogen or chlorine.

8. The following compounds, or a pharmaceutically acceptable salt, or tautomer thereof:

9. a process for the preparation of a compound as claimed in any one of claims 1 to 8, which comprises:

step 1:

step 2:

and the following step 3-1 or step 3-2:

step 3-1:

step 3-2:

wherein R is₀And R₀' are each independently halogen; ring B, R₁、R₂、R₃、R₄、R₅、R₆、R₇X, y are as described in claim 1;

optionally, when R₆Selected from hydrogen, the method comprises any one of step 3-1 or step 3-2;

optionally, when R₆When selected from hydrogen, the process comprises step 3-1; and when R is₆When selected from optionally substituted alkyl, the process comprises step 3-2;

optionally, step 1 is carried out in the presence of a catalyst; optionally, step 1 is carried out in the presence of a catalyst selected from the group consisting of: pd (PPh)₃)₄、PdCl₂(PPh₃)₂、PdCl₂(PhCN)₂、Pd(OAc)₂、Pd/C、PdCl₂(dppf), or a combination thereof; optionally, the catalyst in step 1 is PdCl₂(dppf)；

Optionally, step 1 is carried out in the presence of a base; optionally, step 1 is carried out in the presence of a base selected from: c_SF、K₂CO₃、K₃PO₄Or a combination thereof; optionally, the base in step 1 is cesium fluoride;

optionally, step 3-2 is performed in the presence of a base; optionally, said step 3-2 is performed at a step selected from EtN (i-Pr)₂In the presence of a base of (a);

optionally, step 3-2 is carried out in the presence of a solvent; optionally, said step 3-2 is carried out in the presence of a solvent selected from DCM;

optionally, the method further comprises the following step 1 a:

step 1 a:

optionally, wherein R is₀' is I, step 1a in NIS and HBF₄In the presence of (a).

10. A pharmaceutical composition comprising a compound of any one of claims 1-8, a pharmaceutically acceptable salt, or a tautomer thereof;

optionally, the pharmaceutical composition further comprises pharmaceutically acceptable excipients;

optionally, the pharmaceutical composition further comprises one or more additional therapeutic agents;

optionally, the additional therapeutic agent is an anti-tumor drug;

optionally, the antineoplastic drug is selected from fluorouracil drugs;

optionally, the antineoplastic agent is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouracil, carmofur, doxifluridine, uracil tegafur, capecitabine, tegafur, Youfodin, pyrifudin, fluocinolone, and voroda.

11. Use of a compound of any one of claims 1-8, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of claim 10, or a combination thereof with one or more other therapeutic agents, in the manufacture of a medicament for inhibiting the expression of DPD; or in the manufacture of a medicament for the prevention and/or treatment of a disease, disorder or condition that benefits from inhibition of DPD expression; or in the preparation of medicaments for preventing and/or treating tumors; or in the preparation of medicaments for resisting and/or reversing the drug resistance of fluorouracil medicaments;

optionally, the disease, disorder or condition benefiting from inhibition of DPD expression is selected from the group consisting of a tumor;

optionally, the disease, disorder or condition or the tumor is selected from a tumor of the digestive tract, a tumor of the head and neck, lung cancer, breast cancer; optionally, the head and neck tumor is selected from oral cancer;

optionally, the tumor is a tumor that is resistant to fluorouracil;

optionally, the one or more additional therapeutic agents is an anti-neoplastic drug;

optionally, the antineoplastic drug is selected from fluorouracil drugs;

optionally, the fluorouracil-based drug is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouracil, carmofur, doxifluridine, uracil tegafur, capecitabine, tegafur, Youfodin, pyrifudin, flutolterone, and voroda.

Use of an inhibitor of S1PR2, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition thereof, or a combination thereof with one or more other therapeutic agents, in the manufacture of a medicament for inhibiting the expression of DPD; or in the manufacture of a medicament for the prevention and/or treatment of a disease, disorder or condition that benefits from inhibition of DPD expression; or in the preparation of a medicament for preventing and/or treating tumors, wherein the tumors have drug resistance to fluorouracil drugs; or in the preparation of medicaments for resisting and/or reversing the drug resistance of fluorouracil medicaments;

optionally, the disease, disorder or condition or the tumor is selected from a tumor of the digestive tract, a tumor of the head and neck, lung cancer, breast cancer; optionally, the head and neck tumor is selected from oral cancer; optionally, the disease, disorder or condition benefiting from inhibition of DPD expression is selected from tumours resistant to fluorouracil class drugs;

optionally, the antineoplastic drug is selected from fluorouracil drugs;

optionally, the fluorouracil-based drug is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouracil, carmofur, doxifluridine, tegafur/uracil, capecitabine, tegafur, Youfodin, pyrifudin, flutolterone and voroda.

13. The use according to claim 12, wherein,

the S1PR2 inhibitor is a compound having a structure represented by formula II or formula III, or a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition thereof, or a combination thereof with one or more other therapeutic agents:

in said formula II:

w is-NR^a—、O、—CO—、—CONH—、—NHCH₂-or-CR^bR^c—；

Z is-C (═ O) -, -C (═ S) -, O, — CR^bR^c—、═N—、═CH—

x is a direct bond, -NR^a-, -N ═, -C (═ O) -, -CH ═, O or-CR^bR^c—；

Wherein R is^a、R^bAnd R^cEach independently selected from H or optionally substituted alkyl;

in said formula III:

R²represents (1) a hydrogen atom, (2) an optionally substituted alkyl group;

[ formula 2 ]]

In formula 2, A represents (1) a chemical bond, or (2) an oxygen atom; r¹²And R¹³Each independently represents (1) a hydrogen atom, (2) an optionally substituted alkyl group, (3) a hydroxyl group, or (4) NH₂Or (5) R¹²And R¹³Together with the carbon atom to which they are bonded form C_3-7A carbocyclic ring; the right arrow is bonded to-CONR⁶R⁷、-SO₂R⁸or-COOR⁹，

R⁸represents (1) optionally substituted alkyl, or (2) NR¹⁰R¹¹；

E represents optionally substituted alkyl or

Ring 1 and ring 2 each independently represent a 5-12 membered ring group;

R¹⁴represents (1) a hydrogen atom, or (2) a hydroxyl group;

n represents an integer of 1 to 2;

m represents an integer of 1 to 2;

p represents an integer of 0 to 5;

q represents an integer of 0 to 5;

r represents an integer of 0 to 4;

t represents an integer of 1 to 4;

when p is 2 or more, plural R³May be the same or different;

when q is 2 or more, plural R⁴May be the same or different;

when R is 2 or more, plural R⁵May be the same or different;

14. The use according to claim 13, wherein,

in said formula II:

w is-NR^a—、O、—CO—、—CONH—、—NHCH₂-or-CR^bR^c—；

Z is-C (O) -, -C (S) -, C (S) -and C (S) -are independently substituted,

Y is-NR^a-, O, or-CR^bR^cA; and

x is a direct bond, -NR^a-, - (C) (O) -, O or-CR^bR^c—；

Wherein R is^a、R^bAnd R^cEach independently selected from H or optionally substituted C_1-6An alkyl group;

and/or

In said formula III:

[ formula 2 ]]

In formula 2, A represents (1) a chemical bond, or (2) an oxygen atom; r¹²And R¹³Each independently represents (1) a hydrogen atom, (2) C_1-4Alkyl, (3) hydroxy, or (4) NH₂Or (5) R¹²And R¹³Together with the carbon atom to which they are bonded form C_3-7A carbocyclic ring; the right arrow is bonded to-CONR⁶R⁷、-SO₂R⁸or-COOR⁹，

R⁸represents (1) C_1-4Alkyl group, (2) C_1-4Haloalkyl, or (3) NR¹⁰R¹¹；

R⁹Represents (1) a hydrogen atom, or (2) C_1-8An alkyl group;

E represents alkyl optionally substituted by halogen or

Ring 1 and ring 2 each independently represent a 5-7 membered cyclic group;

R¹⁴represents (1) a hydrogen atom, or (2) a hydroxyl group;

R¹⁷represents (1) C_1-4Alkyl, or (2) a 5-7 membered ring group;

n represents an integer of 1 to 2;

m represents an integer of 1 to 2;

p represents an integer of 0 to 5;

q represents an integer of 0 to 5;

r represents an integer of 0 to 4;

t represents an integer of 1 to 4;

when p is 2 or more, plural R³May be the same or different;

when q is 2 or more, plural R⁴May be the same or different;

when R is 2 or more, plural R⁵May be the same or different;

15. The use according to claim 13 or 14,

in said formula II:

optionally, Ar¹Is optionally substituted aryl or optionally substituted heteroaryl;

optionally, Ar¹Is aryl or heteroaryl, each independently optionally substituted with s groups independently selected from: halogen; optionally substituted alkyl; optionally substitutedAlkoxy group of (a); optionally substituted cycloalkyl; optionally substituted heterocycloalkyl; optionally substituted-O-cycloalkyl; optionally substituted-O-heterocycloalkyl; -SO₂-an alkyl group; -SO₂NR^15aR^15b；-C(＝O)NR^15cR^15d(ii) a An optionally substituted amino group; a hydroxyl group; a cyano group; and a carboxyl group; s represents 0, 1,2,3 or 4;

optionally, Ar¹Is aryl or heteroaryl, each independently optionally substituted with s groups independently selected from: halogen; -C_1-6Alkyl optionally substituted by one or more independently selected R¹³Substituted by groups; -C_1-6Alkoxy optionally substituted by one or more independently selected R¹³Substituted by groups; -C_3-7Monocyclic cycloalkyl optionally substituted with one or more independently selected R¹⁴Substituted by groups; a 4 to 11 membered monocyclic or fused or spirobicyclic heterocycloalkyl containing one, two or three heteroatoms independently selected from N, O and S, optionally substituted with one or more independently selected R¹⁴Substituted by groups; -O-C_3-7Monocyclic cycloalkyl optionally substituted with one or more independently selected R¹⁴Substituted by groups; -O-heterocycloalkyl, wherein the heterocycloalkyl is a 4 to 7 membered monocyclic heterocycloalkyl containing one, two or three heteroatoms independently selected from N, O and S, optionally substituted with one or more independently selected R¹⁴Substituted by groups; -SO₂-C_1-6An alkyl group; -SO₂NR^15aR^15b；-C(＝O)NR^15cR^15d；-NR^17aR^17b(ii) a A hydroxyl group; a cyano group; and a carboxyl group;

wherein each R is¹³Independently selected from: halogen; -CN; -OH; -C_1-4Alkoxy optionally substituted by one or more independently selected OH, C_1-4Alkoxy or halogen substitution; -C (═ O) NR^16aR^16b；-NR^16cC(＝O)-C_1-4An alkyl group; -NR^16dC(＝O)-C_1-4An alkoxy group; -SO₂-C_1-4An alkyl group; -SO₂NR^16eR^16f；-NR^16gSO₂-C_1-4An alkyl group; -C_3-7Monocyclic cycloalkyl optionally substituted by one or moreStereoselective halogen or C optionally substituted by one or more halogens_1-4Alkyl substitution; and 4 to 7 membered monocyclic heterocycloalkyl comprising one, two or three heteroatoms independently selected from N, O and S, optionally substituted with one or more independently selected halogen or C optionally substituted with one or more halogen_1-4Alkyl substitution;

each R^17aAnd R^17bIndependently selected from H and optionally one or more independently selected halogen, OH or C_1-4Alkoxy-substituted C_1-4An alkyl group;

optionally, Ar²Is with Ar¹The same monocyclic heteroaryl group is represented by one of the following structures (1) to (3):

wherein in the structure (1):

R₁is hydrogen or optionally substituted alkyl;

R₃and R₄Each independently selected from hydrogen, deuterium, halogen, -CN, -NO₂Nitroso group, carboxyl group, sulfonic acid group, optionally substituted sulfonyl group, optionally substituted alkyl group, optionally substituted alkoxy group, optionally substituted amino group, optionally substituted cycloalkyl group, optionally substituted cycloalkoxy groupSubstituted aryl or optionally substituted heteroaryl;

wherein in the structure (2):

R³is optionally substituted alkyl;

x is selected from O, NR⁴And CR⁴R⁵Wherein R is⁴、R⁵Each independently selected from the group consisting of H, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted alkoxy, halogen, hydroxy, nitrile, and carboxy;

y is selected from O or S; and

z is an optionally substituted aromatic ring;

wherein in the structure (3):

R^1aselected from:

-NR^6aR^6b；

-C_1-4an alkoxy group;

-C_3-7a monocyclic cycloalkyl group;

-O-C_3-7monocyclic cycloalkyl, and

R^1bis H or C_1-4An alkyl group;

R³selected from:

-C_1-4Alkoxy substituted with one or more halogens;

-C_3-7a monocyclic cycloalkyl group;

-CN；

-S(O)₂-C_1-4an alkyl group;

-NR^8aR^8b(ii) a And

-C(＝O)NR^8cR^8d；

subscript n is 0, 1,2, or 3;

optionally, when Ar is²In the case of the above structure (3), X is-CR^bR^cA; y and W are-NR^a-Z is-C (o) -or-C (═ N-CN) -;

wherein, in said formula III:

optionally, R¹Represents (1) optionally substituted by 1-5R²¹Substituted C_1-8Alkyl, or (2) optionally substituted by 1-5 substituents selected from C_1-4Alkyl radical, C_1-4C substituted by substituent groups of alkoxy, halogen atom and trifluoromethyl_3-7A carbocyclic ring;

optionally, R¹Is C_1-8An alkyl group; optionally R¹Is C_1-6An alkyl group; optionally R¹Is (CH)₃)₂CHCH₂-or (CH)₃CH₂)₂CH-；

Optionally, R²Is a hydrogen atom;

optionally, R¹⁴Is a hydroxyl group;

optionally, M¹And M²Each independently represents-O-or-S-;

optionally, R³And R⁴Each independently represents (1) a halogen atom, (2) C optionally substituted by halogen_1-4Alkoxy, or (3) -L-SO₂R⁸；

Optionally, L represents a bond;

optionally, R⁸Is represented by C_1-4An alkyl group; optionally R⁸Is methyl;

optionally, R³And R⁴Each independently represents F or-SO₂CH₃or-OCH₂CH₂F；

Optionally, ring 1 and ring 2 each independently represent a 5-7 membered ring group selected from: aryl, cycloalkyl or heteroaryl; optionally, ring 1 and ring 2 each independently represent an aryl group; optionally, ring 1 and ring 2 each independently represent phenyl, cyclohexyl, pyridyl; optionally, ring 1 and ring 2 are each independently phenyl;

optionally, E is trifluoromethyl;

optionally, m and n are both 2.

16. Use according to claim 15, wherein

In said formula II:

optionally, R^a、R^bAnd R^cEach independently selected from: hydrogen; or C optionally substituted by a group selected from hydroxy, optionally substituted_1-4C substituted by alkoxy, optionally substituted ester, optionally substituted aminocarbonyl, or carboxyl groups_1-6An alkyl group;

optionally, R^a、R^bAnd R^cEach independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl, or carboxy_1-6An alkyl group;

optionally, R^a、R^bAnd R^cEach independently selected from: hydrogen, methyl, ethyl, -CH₂COOH、-CH₂CH₂OH、-CH₂CH₂CH₂OH、-CH₂CONH₂Or CH₂COOEt；

Optionally, R^a、R^bAnd R^cEach independently selected from hydrogen or methyl；

Optionally, W is-NR^a—；

Optionally, Z is-C (O) -,

optionally, Y is-NR^aA; and

optionally, X is a direct bond, -NR^a-, or-CR^bR^c—；

Optionally, the step of (a) is carried out,

Optionally, in Ar¹Optionally substituted aryl appearing in the definition of (1) is optionally substituted C₆-C₁₀An aryl group;

optionally, in Ar¹Optionally substituted heteroaryl groups appearing in the definition of (a) are optionally substituted monocyclic or fused bicyclic heteroaryl groups;

optionally, in Ar¹Optionally substituted heteroaryl as appearing in the definition of (a) is an optionally substituted monocyclic 5-10 or 5-to 7-membered heteroaryl or an optionally substituted fused bicyclic 8-to 12-membered heteroaryl;

optionally, Ar¹Is optionally substituted phenyl or optionally substituted 5 to 6 membered monocyclic heteroaryl comprising one or two heteroatoms independently selected from N, O and S;

optionally, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; alkyl optionally substituted with halogen, hydroxy or alkoxy; alkoxy optionally substituted with halogen, hydroxy or alkoxy; or amino optionally substituted with alkyl;

optionally, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; optionally substituted by halogen, hydroxy or C_1-6Alkoxy-substituted C_1-6An alkyl group; optionally substituted by halogen, hydroxy or C_1-6Alkoxy-substituted C_1-6An alkoxy group; or optionally is covered with C_1-6An alkyl-substituted amino group;

optionally, optionally，Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; c optionally substituted by halogen, hydroxy_1-6An alkyl group; optionally substituted by hydroxy, C_1-6Alkoxy-substituted C_1-6An alkoxy group; or optionally is covered with C_1-6An alkyl-substituted amino group;

optionally, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; c optionally substituted by halogen_1-6An alkyl group; optionally substituted by hydroxy, or C_1-6Alkoxy-substituted C_1-6An alkoxy group;

optionally, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen, fluorine, chlorine, bromine, trifluoromethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH₂CH₂OCH₃，-OCH₂CH(OH)CH₂OH, N-ethyl-amino or N-ethyl-N-methyl-amino;

optionally, Ar¹Is heteroaryl optionally substituted with s groups independently selected from: hydrogen, chlorine, bromine, trifluoromethyl, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH₂CH(OH)CH₂OH or-OCH₂CH₂OCH₃；

Optionally, in Ar¹Said heteroaryl group as appearing in the definition of (1) is optionally substituted with 1 to 2 of said substituents in ortho position to the heteroatom;

optionally, in Ar¹The heteroaryl group appearing in the definition of (1) is pyridine or thiophene;

optionally, Ar¹Selected from the group consisting of 4-substituted 2, 6-dichloropyridine, 4-substituted 2, 6-dibromopyridine, 4-substituted 2-chloro-6-hydroxyethylpyridine, 4-substituted-2-chloro-6-hydroxypropylpyridine, 4-substituted 2-chloro-6-methoxypyridine, 4-substituted 2-chloro-6-ethoxypyridine, 4-substituted-2-chloro-6-n-propoxypyridine, 4-substituted-2-chloro-6-isopropoxypyridine, 4-substituted-2-chloro-6-n-butoxypyridine, 4-substituted-2-chloro-6- (2-methoxyethoxy) pyridine, and mixtures thereof, 4-substituted-2-trifluoromethyl-6- (2-methoxyethoxy) pyridine, 4-substituted-2-trifluoromethyl6-ethoxypyridine, 4-substituted-2-trifluoromethyl-6- (2, 3-dihydroxypropoxy) pyridine, 4-substituted-2- (N-ethylamino) -6-chloropyridine, 4-substituted-2- (N-ethyl-N-methylamino) -6-chloropyridine, or 5-substituted-2, 3-dichlorothiophene;

optionally, Ar¹Is 4-substituted 2, 6-dichloropyridine, 4-substituted-2-chloro-6- (2-methoxyethoxy) pyridine, 4-substituted-2-trifluoromethyl-6-ethoxypyridine or 4-substituted-2-trifluoromethyl-6- (2, 3-dihydroxypropoxy) pyridine;

optionally, Ar¹And Ar²Are each 4-substituted 2, 6-dichloropyridines;

optionally, Ar¹Is aryl optionally substituted with s groups independently selected from: hydrogen, halogen, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, optionally substituted cycloalkoxy or optionally substituted cycloalkyl;

optionally, Ar¹Is aryl optionally substituted with s groups independently selected from: hydrogen, halogen, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Haloalkyl, C_1-6Haloalkoxy, amino, mono-or di-C_1-6Alkylamino radical, C_3-7Cycloalkyl or C_3-7A cycloalkoxy group;

optionally, Ar¹Is aryl optionally substituted with s groups independently selected from: hydrogen, halogen, C_1-6Alkyl radical, C_1-4Alkoxy radical, C_1-6Perhaloalkyl, C_1-4Perhaloalkoxy, amino, mono-or di-C_1-6Alkylamino radical, C_3-7Cycloalkyl or C_3-7A cycloalkoxy group;

wherein, in the structure (1) of the formula II:

optionally, R₁Is optionally substituted C_1-12An alkyl group; optionally, R₁Selected from C optionally substituted by halogen, alkenyl or hydroxy_1-6An alkyl group; optionally, R₁Selected from C optionally substituted by alkenyl or hydroxy_1-6An alkyl group; optionally, R₁Is methyl, ethyl, n-propyl or isopropylRadical, allyl or-CH₂CH₂CH₂OH; optionally, R₁Is methyl, allyl or-CH₂CH₂CH₂OH; optionally, R₁Is methyl;

optionally, R₂Selected from hydrogen, deuterium, halogen, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Perhaloalkyl radical, C_1-6Perhaloalkoxy, amino, mono-or di-C_1-6Alkylamino radical, C_3-7Cycloalkyl or C_3-7A cycloalkoxy group; optionally, R₂Selected from hydrogen and C_1-6An alkyl group; optionally, R₂Selected from hydrogen and methyl;

optionally, R₃And R₄Each independently selected from hydrogen, deuterium, halogen, -CN, -NO₂Nitroso group, carboxyl group, sulfonic acid group, optionally substituted sulfonyl group, optionally substituted C_1-6Alkyl, optionally substituted C_1-6Alkoxy, optionally substituted amino, optionally substituted C_3-7Cycloalkyl, optionally substituted C_3-7Cycloalkoxy, optionally substituted C₆-C₁₈Aryl or optionally substituted C₁-C₁₈A heteroaryl group;

optionally, R₃And R₄Each independently selected from hydrogen; and C independently of one another optionally substituted by_1-6Alkyl radical, C_3-7Cycloalkyl radical, C₆-C₁₈Aryl or C₁-C₁₈Heteroaryl group: halogen, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl or C_1-6An alkoxy group;

optionally, R₃And R₄Each independently selected from: hydrogen; c_3-7A cycloalkyl group; optionally substituted by halogen, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl or C_1-6Alkoxy-substituted C_1-6An alkyl group; each independently optionally substituted by halogen, or C_1-6Alkoxy-substituted C₆-C₁₈Aryl or C₁-C₁₈A heteroaryl group;

optionally, R₃And R₄Each independently selected from: hydrogen;C_3-7a cycloalkyl group; optionally substituted by halogen or C₆-C₁₈Aryl substituted C_1-6An alkyl group; optionally substituted by halogen, or C_1-6Alkoxy-substituted C₆-C₁₈An aryl group; or C optionally substituted by halogen₁-C₁₈A heteroaryl group;

optionally, R₃And R₄Each independently selected from: hydrogen; c_3-6A cycloalkyl group; c optionally substituted by halogen or phenyl_1-6An alkyl group; optionally substituted by halogen, or C_1-6Alkoxy-substituted phenyl; or furyl or thienyl each independently optionally substituted with halogen;

optionally, R₃And R₄Each independently selected from: hydrogen, methyl, n-propyl, isopropyl, cyclopropyl, cyclohexyl, -CF₃Benzyl, phenyl, fluoro-substituted phenyl, methoxy-substituted phenyl, furyl or chloro-substituted thienyl;

optionally, R₃And R₄Is not hydrogen and is located at h;

wherein, in the structure (2) of the formula II:

optionally, the structure (2) is represented by the following structure (2-1):

d is CR or N;

x is O, NR⁴，CR⁴R⁵Wherein X is as definedR appearing in⁴And R⁵Independently selected from: h and alkyl groups such as lower alkyl groups and branched alkyl groups which may have 1 to 10 carbons and may be cyclic or branched alkyl groups having 3 to 10 carbons; a methoxy group; a hydroxyl group; f; br; i; a nitrile; trifluoromethyl and carboxyl;

y is O or S;

z is a substituted aromatic ring having the structure:

e is N or CR;

optionally, the structure (2) is represented by the following structure (2-2):

y is O or S;

z is a substituted aromatic ring having the structure:

e is N or CR;

wherein, in the structure (3) of the formula II:

optionally, R^1aIs selected from methyl;

optionally, R^1bSelected from hydrogen;

optionally, the step of (a) is carried out,

is composed of

Optionally, R³And R⁴Each independently selected from hydrogen or C optionally substituted by halogen_1-6An alkyl group;

optionally, R³Is methyl;

optionally, R⁴Is hydrogen or trifluoromethyl; optionally, R⁴At least one of which is trifluoromethyl.

17. The use according to claim 12, wherein the composition is administered to a mammal,

optionally, the S1PR2 inhibitor is a compound, or a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition thereof, or a combination thereof with one or more other therapeutic agents:

18. a method for inhibiting DPD expression in a cell, comprising administering to the cell an effective amount of a compound of any one of claims 1-8, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of claim 10, or combinations thereof with one or more other therapeutic agents, or an inhibitor of S1PR2 of any one of claims 12 to 17, wherein the method is performed in vitro;

wherein optionally, the cell is a cell line, or a cell from a subject;

optionally, the cell is selected from a tumor cell; optionally, the tumor cell is selected from a breast cancer cell, a liver cancer cell, a lung cancer cell, an esophageal cancer cell, a stomach cancer cell, a pancreatic cancer cell, a bile duct cancer cell, a head and neck tumor cell, or a nasopharyngeal cancer cell.

19. A method for combating/reversing drug resistance in a cell, comprising administering to said cell an effective amount of a compound of any one of claims 1-8, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of claim 10, or a combination thereof with one or more other therapeutic agents, or an S1PR2 inhibitor of any one of claims 12-17, wherein said method is performed in vitro;

wherein optionally, the cell is a cell line, or a cell from a subject;

20. A method of screening for an anti-drug resistant oncology therapeutic, wherein the anti-drug resistant oncology therapeutic is an oncology therapeutic capable of resisting and/or reversing drug resistance to fluorouracil, the method comprising:

providing a tumor cell line or cell culture expressing S1PR 2;

contacting a candidate with the tumor cell line or cell culture;

21. A method of screening for an anti-drug resistant oncology therapeutic, wherein the anti-drug resistant oncology therapeutic is an oncology therapeutic capable of resisting and/or reversing drug resistance to fluorouracil, comprising:

providing a DPD-expressing tumor cell line or cancer cell culture;

contacting a candidate with the tumor cell line or cancer cell culture;

wherein the candidate is selected from S1PR2 inhibitors.

22. A kit, comprising:

a compound of any one of claims 1-8, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of claim 10, or an S1PR2 inhibitor of any one of claims 12-17;

optionally an anti-tumour drug, optionally selected from fluorouracil drugs;

and optionally further including instructions for use.