CN113135909B

CN113135909B - DPD inhibitor, preparation method thereof, pharmaceutical composition and application

Info

Publication number: CN113135909B
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: 2024-03-19
Anticipated expiration: 2040-01-17
Also published as: CN113135909A

Abstract

The application relates to a novel DPD expression inhibitor, a preparation method, a pharmaceutical composition and application thereof, and a novel application of a compound such as an S1PR2 inhibitor, in particular to an application of the S1PR2 inhibitor in inhibiting the expression of dihydropyrimidine dehydrogenase DPD, in particular to an application in reversing drug resistance when fluorouracil drugs treat tumors.

Description

DPD inhibitor, preparation method thereof, pharmaceutical composition and application

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 a novel application of a compound of a sphingosine-1-phosphate receptor (S1 PR 2) inhibitor. In particular to the use of an S1PR2 inhibitor for inhibiting the expression of dihydropyrimidine dehydrogenase DPD.

Background

Chemotherapy plays an extremely important role in the treatment of cancer. Since 1957Duschinsky et al synthesized 5-fluorouracil, 5-fluorouracil and its derivatives have been the basis for chemotherapy of digestive tract tumors, head and neck tumors, and other tumors, such as lung and breast cancers. However, as with other anticancer drugs, the development of resistance to tumors is a major cause of failure of the above-described cancer chemotherapy. In fact, the intracellular dihydropyrimidine dehydrogenase (DPD) is the main 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 not known at present, so that the discovery of a new target point is not in a talk with the targeted design of a 5-fluorouracil drug resistance reversal agent, which is also exactly the urgent need of a clinical doctor of the seesaw. Therefore, it is important to identify key molecules involved in the regulation of DPD expression during the development of 5-fluorouracil resistance and to identify them as targets for designing drug resistance reversal agents.

Sphingosine-1-phosphate subtype receptors (S1 PR2, also known as EDG-5, H218, AGR16, lpB 2) are typical members of the rhodopsin (rhodopsin-like receptor, class A) family as one of the G protein coupled receptor family members. The main functions reported in the current research are inhibiting endothelial cell migration, promoting insulin content, inhibiting tumor cell migration and the like. The FTY720 applied in clinic can be combined with other targets S1PR1, S1PR3, S1PR4 and S1PR5 on the surface of inflammatory cell membranes in a phosphorylation mode through simulating the S1P structure, so that corresponding functions of inhibiting inflammatory factors are generated, but the FTY720 is only not combined with the S1PR2 or combined very poorly, and downstream effective signal transmission cannot be caused.

Currently available S1PR2 inhibitors are mainly pyridopyrazoles, diaryloxyphenyl, pyridazines, such as those described in WO08154470, WO 2011/04287, WO2011/159864, WO2013/148460, WO2014/158302, WO2016/191872, US 2019/0127172, 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) 4387-4392, bioorg.med.chem.lett.26 (2016) 1209-1213, bioorg.med.chem.lett.28 (2018) 488-496, etc., which have no effect of inhibiting DPD expression of cancer cells. The above-mentioned patent and non-patent documents are incorporated herein by reference 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:

R ₁ selected from optionally substituted aryl, optionally substituted heteroaryl or optionally substituted heterocyclenyl;

R ₂ selected from hydrogen or optionally substituted alkyl;

each R is ₃ Independently selected from hydrogen, deuterium, halogen, -CN, -NO ₂ A nitroso group, a carboxyl group, a sulfonic group, an optionally substituted sulfonyl group, an optionally substituted alkyl group, an optionally substituted cycloalkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group;

R ₄ 、R ₅ and R is ₆ Each independently selected from hydrogen, or optionally substituted alkyl;

ring B is an aromatic or heteroaromatic ring;

each R is ₇ Independently 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 pharmaceutically acceptable salts, or tautomers thereof:

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

step 2:

the following steps 3-1 or 3-2:

step 3-1:

step 3-2:

wherein R is ₀ And R is ₀ ' each independently is halogen; ring B, R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ X, and y are as described in the first aspect above.

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

In a fifth aspect, the present application provides the use of a compound according to the first aspect, a pharmaceutically acceptable salt, or a tautomer thereof, or a pharmaceutical composition according to the fourth aspect, or a combination of same and one or more other therapeutic agents, for 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 benefiting from inhibition of DPD expression; or in the preparation of a medicament for preventing and/or treating tumors; or in the preparation of a medicament for resisting and/or reversing the resistance of fluorouracil medicaments.

In a sixth aspect, the present application provides the use of an S1PR2 inhibitor, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition thereof, or a combination of same 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 benefiting from inhibition of DPD expression; or in the manufacture of a medicament for the prevention and/or treatment of a tumor, wherein the tumor is resistant to fluorouracil; or in the preparation of a medicament for resisting and/or reversing the resistance of fluorouracil medicaments.

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

In an eighth aspect, the present application provides a method for combating/reversing drug resistance of a cell, comprising administering to said cell an effective amount of 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 of same with one or more other therapeutic agents, or an S1PR2 inhibitor of the sixth aspect above, wherein said method is performed in vitro.

In a ninth aspect, the present application provides a method of screening for a drug resistant tumor therapeutic agent, wherein the drug resistant tumor therapeutic agent is a tumor therapeutic agent capable of resisting and/or reversing fluorouracil drug resistance, 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 level of expression of S1PR2 in the tumor cell line or cell culture after contact with the candidate and comparing the level of expression of S1PR2 with a control tumor cell line or cell culture not contacted with the candidate;

when the S1PR2 expression level is lower than the control tumor cell line or cell culture, the candidate is indicated to be capable of acting as an anti-drug resistant tumor therapeutic.

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

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

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

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

when DPD expression levels are lower than control tumor cell lines or cell cultures, the candidate is indicated to be capable of acting as an anti-drug resistant tumor therapeutic;

Wherein the candidate is selected from the group consisting of S1PR2 inhibitors.

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

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

optionally an antitumor drug.

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

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

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

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) in need thereof a therapeutically effective amount of a compound of the above first aspect, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of the above fourth aspect, or a combination of same with one or more additional therapeutic agents, or an S1PR2 inhibitor of the above sixth aspect.

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

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

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

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

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

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

Drawings

Fig. 1 shows experimental results of the inhibition of DPD expression in vitro of existing S1PR2 inhibitors and newly synthesized compounds of the present disclosure.

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

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

FIG. 4 shows the inhibition of DPD expression in vitro after S1PR2 knockdown.

Detailed Description

Definition of the definition

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 construed according to conventional 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 english variants such as comprises or comprising should be understood in an open, non-exclusive sense, i.e. "including but not limited to".

The term "optional" or "optionally" means 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.

The numerical ranges herein indicating the number of carbon atoms refer to each integer in the given range, e.g., "C ₁ -C ₁₈ "OR" C _1-18 By "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" refers to the number of backbone atoms that make up the ring. For example, "3 to 12 membered" means that the number of skeleton atoms constituting a ring is 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

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

The term "alkyl" refers to a straight or branched chain saturated hydrocarbon group that may be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). Examples of alkyl groups include C _1-4 Alkyl, C _1-6 Alkyl, C _1-8 Alkyl, C _1-10 Alkyl, C _1-12 Alkyl groups and the like, such as 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 _1-6 Alkyl "refers to an alkyl group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.) containing from 1 to 6 (e.g., 1, 2, 3, 4, 5, 6) carbon atoms.

The term "hydroxyalkyl" refers to an-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 an-O-alkyl group, wherein "alkyl" is 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, e.g. C _2-6 Alkenyl, C _2-4 Alkenyl groups, and the like. Non-limiting examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, 1, 3-butadienyl, and the like. The term includes both cis and trans isomers or mixtures of these isomers.

The term "alkynyl" refers to a straight or branched chain unsaturated aliphatic hydrocarbon group consisting of carbon and hydrogen atoms, having at least one triple bond, e.g. C _2-6 Alkynyl, C _2-4 Alkynyl, C _2-3 Alkynyl groups, and the like. Non-limiting examples of alkynyl groups include, but are not limited to, ethynyl (-C.ident.CH), 1-propynyl (-C.ident.C-CH) ₃ ) 2-propynyl (-CH) ₂ -C.ident.CH), 1, 3-butadienyl (-C.ident.C-C.ident.CH), and the like.

The term "cycloalkyl" refers to a fully saturated non-aromatic ring consisting 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 groups include, but are not limited to, C _3-18 Cycloalkyl, C _3-16 Cycloalkyl, C _3-12 Cycloalkyl, C _3-10 Cycloalkyl, C _3-8 Cycloalkyl, C _3-7 Cycloalkyl, C _3-6 Cycloalkyl 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, bridged ring, or spiro ring. Unless otherwise indicated, the heterocycle is typically a ring containing from 1 to 5 (e.g., 1, 2, 3, 4, 5) heteroatoms independently selected from sulfur, oxygen, and/or nitrogen. Examples of 3-membered heterocycloalkyl groups include, but are not limited to, ethylene oxide, ethylene nitride, non-limiting examples of 4-membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, examples of 5-membered heterocycloalkyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, isoxazolidinyl, oxazolidinyl, isothiazolidinyl, thiazolidinyl, imidazolidinyl, tetrahydropyrazolyl, examples of 6-membered heterocycloalkyl groups include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, 1, 4-thiaalkyl, 1, 4-dioxanyl, thiomorpholinyl, 1, 3-dithianyl, 1, 4-dithianyl, examples of 7-membered heterocycloalkyl groups include, but are not limited to, azepanyl, oxepinyl, 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, cycloalkenyl rings contain, for example, 3-10 ring atoms, 5-10 ring atoms, or 5-7 ring atoms. Non-limiting examples of suitable monocyclic cycloalkenyl groups include cyclopentenyl, cyclohexenyl, cycloheptenyl, and the like. A non-limiting example of a suitable polycyclic cycloalkenyl group is norbornenyl.

The term "heterocycloalkenyl" refers to a non-aromatic monocyclic or polycyclic ring system which contains at least one carbon-carbon double bond or carbon-nitrogen double bond and which consists solely of carbon and hydrogen atoms, wherein one or more ring atoms in the ring system are elements other than carbon, e.g., 1,2,3,4, 5 ring atoms are each independently heteroatoms selected from nitrogen, oxygen, and sulfur. Adjacent oxygen and/or sulfur atoms are not present in the ring system, and in some embodiments, the heterocycloalkenyl ring contains, for example, 5-10 ring atoms, 5-8 ring atoms, 5-7 ring atoms, or 5-6 ring atoms. The pre-conjugated aza, oxa or thia before the heterocycloalkenyl name indicates that at least one nitrogen, oxygen or sulfur atom, respectively, is present as a ring atom. Non-limiting examples of suitable monocyclic azetidinyl groups include 1,2,3, 4-tetrahydropyridine, 1, 2-dihydropyridinyl, 1, 4-dihydropyridinyl, 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, dihydrofuryl, fluorodihydrofuryl, and the like. A non-limiting example of a suitable polyepoxide group is 7-oxabicyclo [2.2.1] heptene group.

The term "cycloalkynyl" refers to a non-aromatic mono-or polycyclic ring having at least one carbon-carbon triple bond and consisting of only carbon and hydrogen atoms, e.g., a 4-15 membered, 5-15 membered, 6-10 membered, 7-10 membered, or 8-10 membered ring, such as an 8-to 10-membered mono-ring or a 12-to 15-membered bi-ring. Which may comprise one or more fused or bridged rings. Unless otherwise indicated, 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, cyclodeynyl, 2-methylcyclooctynyl, and the like.

The term "heterocycloalkynyl" refers to a heterocycloalkyl group in which at least one single carbon-carbon bond is replaced with a single 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 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 covalently linked. Non-limiting examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthryl, 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 "C ₆ -C ₁₈ Aryl "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 "heteroaryl ring" refers to an aromatic ring group consisting of carbon atoms and at least one (e.g., 1 to 5, such as 1,2, 3, 4, 5) heteroatoms selected from nitrogen, oxygen, and sulfur. Heteroaryl groups may be monocyclic, bicyclic, tricyclic or tetracyclic ring systems, which may include fused or bridged ring systems; and the nitrogen, carbon or sulfur atoms in the heteroaryl group may optionally be oxidized; the nitrogen atom may optionally be quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzothiadiazolyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyronyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo [4,6] imidazo [1,2-a ] pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, furanyl, thienyl, furanonyl, isothiazolyl, imidazolyl indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, phenazinyl, phenothiazinyl, phenoxazinyl, 2, 3-naphthyridinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl.

The term "C ₁ -C ₁₈ Heteroaryl "refers to an aromatic ring group having 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 carbon atoms) selected from N, O, and S as a ring-forming atom 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 which 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 "C ₁ -C ₁₈ Heterocyclyl "means having a ring-forming atom in addition to 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) selected from N, O, andheterocyclyl as defined above of at least one heteroatom (e.g. 1, 2, 3, 4 or 5 heteroatoms) in S.

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

The term "4-7 membered nitrogen containing saturated heterocyclic ring" means that it contains 1-5 (e.g. 1, 2, 3, 4 or 5) heteroatoms selected from oxygen, nitrogen and sulfur, and that the partially or fully saturated 4-7 membered monocyclic heterocyclic ring must contain one or more nitrogen atoms. For example: azetidines, pyrrolines, pyrrolidines, imidazolines, imidazolidines, triazolines, tetrazolines, tetrazoles, pyrazolines, pyrazolidines, dihydropyridines, tetrahydropyridines, piperidines, dihydropyrazines, tetrahydropyrazines, piperazines, dihydropyrimidies, tetrahydropyrimidines, perhydropyrimidines, dihydropyridazines, tetrahydropyridazines, perhydro pyridazines, dihydro aza, tetrahydroaza, perhydro aza, dihydro diaza, tetrahydrodiaza, perhydro diaza, dihydro oxazoles, tetrahydrooxazoles (oxazolidines), dihydro isoxazoles, tetrahydroisoxazoles (isoxazolidines), dihydro thiazolidines, dihydro isothiazoles, tetrahydroisothiazoles (isothiazolidines), dihydrofurazanes, tetrahydrofurazanes, dihydrodiazoles, tetrahydrodiazoles (diazolidines), dihydrooxazines, tetrahydrooxazines, dihydrodiazines, tetrahydrodiazines, dihydrooxazanes, peroxyhydroxazanes, dihydrooxadiazines, peroxyhydroxadiazoles, dihydrothiadiazoles, tetrahydrothiadiazoles (thiadiazolidines), dihydrothiazines, tetrahydrothiazines, dihydrothiadiazines, tetrahydrothiadiazines, perhydro thiadiazines, morpholine or thiomorpholine rings.

A5-12 membered ring group or 5-12 membered ring refers to a C5-12 carbocyclic ring and a 5-12 membered heterocyclic ring. A5-7 membered ring group refers to a C5-7 carbocyclic ring and a 5-7 membered heterocyclic ring. Here, carbocycles and heterocycles have the same meaning as above, and examples of 5-to 7-membered heterocycles include: pyrrolines, pyrrolidines, imidazolines, imidazolidines, triazolines, tetrazolines, tetrazoles, pyrazolines, dihydropyridines, tetrahydropyridines, piperidines, dihydropyrazines, tetrahydropyrazines, piperazines, dihydropyrimidies, tetrahydropyrimidines, perhydro pyrimidines, dihydropyridazines, tetrahydropyridazines, perhydro pyridazines, dihydro aza, tetrahydro aza, perhydro aza, dihydro diaza, perhydro diaza, dihydro furan, tetrahydrofuran, dihydropyran, tetrahydropyran, dihydrooxa, tetrahydrooxa, perhydroxy oxa, dihydro thiophene, tetrahydrothiophene, thiocyran, tetrahydrothiopyran, dihydrothia, tetrahydrothia, perhydro thia, dihydro azole, tetrahydroazole (oxazolidine), dihydro isoxazole, tetrahydroisoxazole (isoxazolidine), dihydro thiazole, tetrahydrothiazole (thiazolidine), dihydro isothiazole, tetrahydrothiopyran, dihydropyran tetrahydroisothiazoles (isothiazolidines), dihydrofurazanes, tetrahydrofurazanes, dihydrodiazoles, tetrahydrodiazoles (diazolidines), dihydrooxazines, tetrahydrooxazines, dihydrodiazines, tetrahydrodiazines, dihydroazenes, tetrahydroazetidines, tetrahydroazepines, dihydroazepines, dihydroazetidines, dihydroazepines, and dihydroazepines, which can be used as a catalyst to treat or to treat a disorder, such as a disorder tetrahydrooxazas, peroxazas, dioxadiaza-diaza, tetraoxadiaza-diaza, peroxa-diaza, dihydrothiadiazoles, tetrahydrothiadiazoles (thiadiazolidines), dihydrothiazines, tetrahydrothiazines dihydrothiadiazines, tetrahydrothiadiazines, dihydrothiaazenes, tetrahydrothiaazenes, perhydrothiaazenes, dihydrothiadiazines, tetrahydrothiadiazines, perhydrothiadiazines, morpholines, thiomorpholines, thiazines, dioxolanes, dioxane, dithiolane, dithianes, pyrrole, imidazole, triazole, tetrazoles, pyrazoles, pyridine, piperazine, pyrimidine, pyridazines, aza, diaza, furan, pyran, oxa, thiophene, thiopyran, thia, azole, isoxazole, thiazole, isothiazole, furazan, diazole, oxazine, diazine, thiadiazole, thiazine, thiadiazine, thiaaza or thiadiazepine.

In the present description of the invention,meaning the connection location.

Double-line consisting of broken line and solid line in this specificationRepresents 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) ₂ CO ₂ H) 2-carboxyethyl, and the like. "optionally substituted carboxyalkyl" means that the alkyl moiety in the group is substituted or unsubstituted.

The term "optionally substituted amino" refers to-NH ₂ Mono-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 ' 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, alkylaryl, alkylheteroaryl. In some embodiments, R' is alkyl or haloalkyl. In some embodiments, R' is C _1-6 An alkyl group. "optionally substituted ester" means 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.

As used herein, the term "sulfonyl" refers to having the meaning-SO ₂ A 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), when substituted, the substituents are independently and independently selected from one or more of the following: 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-amido, N-amido, S-sulfonamido, N-sulfonamido, carboxyl, C-carboxyl, O-carboxyl, isocyanato, thiocyanato, isothiocyanato, nitro, ester, silyl, trihalomethanesulfonyl, amino 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 effectiveness of the free acid and free base of the specified compound and that are biologically or otherwise undesirable. "pharmaceutically acceptable salts" include "pharmaceutically acceptable acid addition salts" and "pharmaceutically acceptable base addition salts".

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

The present application also includes isotopically-labeled compounds identical to those recited herein, but for the replacement of one or more atoms by an atom having an atomic weight or mass number different from the atomic weight 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, respectively ² H、 ³ H、 ¹¹ C、 ¹³ C、 ¹⁴ C、 ¹³ N、 ¹⁵ N、 ¹⁵ O、 ¹⁷ O、 ¹⁸ O、 ³¹ P、 ³² P、 ³⁵ S、 ¹⁸ F、 ¹²³ I、 ¹²⁵ I and ³⁶ cl, and the like.

Certain isotopically-labeled compounds of the present application(e.g. using ³ H is H ¹⁴ C-labeled) can be used in compound and/or substrate tissue distribution analysis. Tritiation (i.e ³ H) And carbon-14 (i.e ¹⁴ C) Isotopes are particularly preferred for their ease of preparation and detectability. Positron emitting isotopes, such as ¹⁵ O、 ¹³ N、 ¹¹ C and C ¹⁸ F 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 examples below by substituting an isotopically-labeled reagent for an non-isotopically-labeled reagent.

In addition, the use of heavier isotopes (such as deuterium (i.e. ² H) Substitution may provide certain therapeutic advantages resulting from higher 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 are included within the scope of the present application.

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

The term "treatment" means administration of 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 a disease or disease state, i.e., inhibiting its progression;

(ii) The disease or condition is alleviated, even if the disease or condition subsides.

The term "preventing" means that a compound or formulation described herein is administered to prevent a disease or one or more symptoms associated with the disease, and includes: preventing a disease or a disease state from occurring in a mammal, particularly when such mammal is susceptible to the disease state, but has not been diagnosed as having the disease state.

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 of a disease, or any other desired alteration of a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is required to provide clinically significant relief from a disorder. Effective amounts suitable in any individual case can be determined using techniques such as a dose escalation test.

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

"pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent or emulsifier, and the like that have been approved by the U.S. food and drug administration for use in humans or animals.

The pharmaceutical compositions of the present application may be prepared by combining the compounds of the present application with suitable pharmaceutically acceptable excipients, for example, in 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 routes of administration of the compounds of the present application, pharmaceutically acceptable salts, tautomers thereof, 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, suspensions for injectable administration; patches for transdermal administration, subcutaneous deposits, and the like. Injections may be prepared in the following conventional forms: solutions or suspensions, solid dosage forms suitable for making solutions or suspensions 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 composition may contain minor amounts of non-toxic auxiliary substances such as wetting agents, pH buffers, and the like. Absorption enhancers (e.g., liposomes) may also be used if desired.

Formulations for parenteral administration include aqueous solutions of the active compound 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, pomelo oil, or almond oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, for example sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may contain suitable stabilizers or agents that enhance the solubility of the compounds, so that high concentration solutions may be prepared.

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

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 excipients or excipients well known in the art. These adjuvants or excipients enable the compounds of the present application to be formulated as tablets, pills, troches, 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 tabletting methods. For example, the oral composition in solid form can be obtained by the following method: the active compound is admixed with solid excipients or excipients, the resulting mixture is optionally milled, if desired with other suitable excipients or excipients, and the mixture is then processed to granules, giving a tablet or dragee core. Suitable excipients or excipients include, but are not limited to: fillers, binders, diluents, disintegrants, lubricants, glidants, sweeteners or flavoring agents, and the like. Pharmaceutical formulations for oral administration, for example, can be obtained by the following method: combining the active compound with a solid excipient, optionally milling the resulting mixture, and processing the mixture of granules, if desired, to give tablets or dragee cores after adding suitable adjuvants. 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, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired, disintegrating agents can be added, for example crosslinked polyvinylpyrrolidone, agar or alginic acid or an alginate such as sodium alginate. Dragee cores are suitably coated. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. To identify or characterize different combinations of active compound doses, dyes or pigments may be added to the tablets or dragee coatings. 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 dosage 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, such as 5-fluorouracil (5-FU), 5'-2' -deoxyuridine, tegafur (tegafur), fluorouridine, carmofur, deoxyfluorouridine, tegafur/uracil, capecitabine (capecitabine), tegafur, eufudine, cyprodinil, fluroxypyr, and the like.

The expression "disease, disorder or condition benefiting 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 nasopharyngeal carcinoma, etc.

wherein:

R ₂ Selected 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, R ₁ Selected from aryl, heteroaryl, or heterocycloalkenyl groups each independently optionally substituted with halogen, -OH, -CN, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl, alkoxy, aryloxy, or heteroaryloxy;

in some embodiments, R ₁ Selected from each independently optionally substituted with halogen, -OH, -CN, C _1-6 Alkyl, C _1-6 Haloalkyl, C _1-6 Hydroxyalkyl, 3-10 membered cycloalkyl, C _1-6 Alkoxy, C ₆ -C ₁₈ Aryloxy or C ₁ -C ₁₈ Heteroaryloxy-substituted C ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl, or 5-10 membered heterocyclenyl;

in some embodiments, R ₁ Selected from each independently optionally substituted with halogen, -OH, -CN, C _1-6 Alkyl, C _1-6 Alkoxy or C ₆ -C ₁₈ Aryloxy substituted C ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl, or 5-10 membered heterocyclenyl;

in some embodiments, R ₁ Selected from C independently optionally substituted with fluorine, chlorine, -OH, -CN, methyl, ethyl, n-propyl, isopropyl, n-propoxy, isopropoxy or phenoxy ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl, or 5-10 membered heterocyclenyl;

in some embodiments, R ₁ Selected from C independently optionally substituted with fluoro, -OH, -CN, methyl, n-propoxy, isopropoxy or phenoxy ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl, or 5-10 membered heterocyclenyl;

in some embodiments, R ₁ Selected from: optionally selected from halogen, -OH, -CN, C _1-6 Alkyl, C _1-6 Alkoxy, or C ₆ -C ₁₈ Group-substituted C of aryloxy group ₆ -C ₁₈ An aryl group; optionally by C _1-6 Alkoxy substituted C ₁ -C ₁₈ Heteroaryl; or a 5-to 10-membered heterocycloalkenyl group;

in some embodiments, R ₁ The C occurring in the definition of (2) ₆ -C ₁₈ Aryl is phenyl;

in some embodiments, R ₁ The C occurring in the definition of (2) ₁ -C ₁₈ Heteroaryl is selected from pyridinyl or furanyl;

in some embodiments, the 5-10 membered heterocycloalkenyl is

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

in some embodiments, R ₁ Selected from: phenyl optionally substituted with a group selected from fluoro, -OH, -CN, methyl, n-propoxy, or phenoxy; isopropoxy substituted pyridinyl; a furyl group; or alternatively/>

In some embodiments, R ₂ Selected from hydrogen, optionally substituted C _1-6 An alkyl group;

in some embodiments, R ₂ Selected from optionally substituted C _1-6 An alkyl group;

in some embodiments, R ₂ Selected from C optionally substituted by halogen, alkenyl or hydroxy _1-6 An alkyl group;

in some embodiments, R ₂ Selected from C optionally substituted by alkenyl or hydroxy _1-6 An alkyl group;

in some embodiments, R ₂ Is methyl, ethyl, n-propyl, isopropyl, allyl or-CH ₂ CH ₂ CH ₂ OH；

In some embodiments, R ₂ Is methyl, allyl or-CH ₂ CH ₂ CH ₂ OH；

In some embodiments, R ₂ Is methyl.

In some embodiments, each R ₃ Independently selected from hydrogen, deuterium, halogen, -CN, -NO ₂ Nitroso, carboxyl, sulfonic acid group, optionally substituted sulfonyl, optionally substituted C _1-6 Alkyl, optionally substituted C _3-8 Cycloalkyl, optionally substituted C ₆ -C ₁₈ Aryl, optionally substituted C ₁ -C ₁₈ Heteroaryl;

in some embodiments, each R ₃ Independently selected from: hydrogen; each independently optionally substituted with halogen, C ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl or C _1-6 Alkoxy substituted C _1-6 Alkyl, C _3-8 Cycloalkyl, C ₆ -C ₁₈ Aryl or C ₁ -C ₁₈ Heteroaryl;

in some embodiments, each R ₃ Independently selected from: hydrogen; c (C) _3-8 Cycloalkyl; optionally by halogen, C ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl or C _1-6 Alkoxy substituted C _1-6 An alkyl group; each independently optionally substituted with halogen, or C _1-6 Alkoxy substituted C ₆ -C ₁₈ Aryl or C ₁ -C ₁₈ Heteroaryl;

in some embodiments, each R ₃ Independently selected from: hydrogen; c (C) _3-8 Cycloalkyl; optionally by halogen or C ₆ -C ₁₈ Aryl substituted C _1-6 An alkyl group; optionally by halogen, or C _1-6 Alkoxy substituted C ₆ -C ₁₈ An aryl group; or C optionally substituted by halogen ₁ -C ₁₈ Heteroaryl;

in some embodiments, each R ₃ Independently selected from: hydrogen; c (C) _3-8 Cycloalkyl; c optionally substituted by halogen or phenyl _1-6 An alkyl group; optionally by halogen, or C _1-6 Alkoxy substituted phenyl; or furyl or thienyl each independently optionally substituted with halogen;

in some embodiments, each R ₃ Independently selected from hydrogen, methyl, n-propyl, isopropyl, cyclopropyl, cyclohexyl, -CF ₃ Benzyl, benzeneA phenyl, fluoro-substituted phenyl, methoxy-substituted phenyl, furyl or chloro-substituted thienyl group;

in some embodiments, each R ₃ Independently selected from hydrogen, or methyl;

in some embodiments, at least one R ₃ Is not hydrogen and is located para to N;

in some embodiments, x is 1.

In some embodiments, R ₄ 、R ₅ And R is ₆ Each independently selected from: hydrogen; or an alkyl group optionally substituted with a group selected from hydroxy, optionally substituted ester, optionally substituted aminocarbonyl, or carboxy;

in some embodiments, R ₄ 、R ₅ And R is ₆ Each independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, optionally substituted ester, optionally substituted aminocarbonyl, or carboxyl _1-6 An alkyl group;

in some embodiments, R ₄ 、R ₅ And R is ₆ Each independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl, or carboxyl _1-6 An alkyl group;

in some embodiments, R ₄ 、R ₅ And R is ₆ Each independently selected from: hydrogen, methyl, ethyl, -CH ₂ COOH、-CH ₂ CH ₂ OH、-CH ₂ CH ₂ CH ₂ OH、-CH ₂ CONH ₂ Or CH (CH) ₂ COOEt；

In some embodiments, R ₄ 、R ₅ And R is ₆ Each independently selected from hydrogen or methyl;

in some embodiments, R ₄ 、R ₅ And R is ₆ Are all hydrogen.

In some embodiments, the heteroaromatic ring appearing in the definition of ring B is C ₁ -C ₁₈ A heteroaromatic ring;

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

in some embodiments, the aromatic ring appearing in the definition of ring B is C ₆ -C ₁₈ An aromatic ring;

in some embodiments, the aromatic ring appearing in the definition of ring B is C ₆ -C ₁₀ An aromatic ring;

in some embodiments, the aromatic ring appearing 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 appearing in the definition of ring B is pyridine or thiophene;

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

in some embodiments, the heteroaromatic ring appearing in the definition of ring B is optionally substituted with 1-2R in the ortho-position to the heteroatom ₇ Group substitution;

in some embodiments of the present invention, in some embodiments,selected from pyridinyl or thienyl optionally substituted with: hydrogen, fluorine, chlorine, bromine, trifluoromethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH ₂ CH ₂ OCH ₃ ，-OCH ₂ CH(OH)CH ₂ OH, N-ethyl-amino or N-ethyl-N-methyl-amino;

in some embodiments of the present invention, in some embodiments,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-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, 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, in some embodiments,is 4-substituted 2, 6-dichloropyridine.

In some embodiments, each R ₇ 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, each R ₇ Independently selected from hydrogen; halogen; optionally by halogen, hydroxy or C _1-6 Alkoxy substituted C _1-6 An alkyl group; optionally by halogen, hydroxy or C _1-6 Alkoxy substituted C _1-6 An alkoxy group; or optionally by C _1-6 An alkyl-substituted amino group;

in some embodiments, each R ₇ Independently selected from hydrogen; halogen; c optionally substituted by halogen or hydroxy _1-6 An alkyl group; optionally by hydroxy or C _1-6 Alkoxy substituted C _1-6 An alkoxy group; or optionally by C _1-6 An alkyl-substituted amino group;

in some embodiments, each R ₇ Independently selected from hydrogen; halogen; c optionally substituted by halogen _1-6 An alkyl group; optionally by hydroxy or C _1-6 Alkoxy substituted C _1-6 An alkoxy group;

in some embodiments, each R ₇ Independently selected from the group consisting of hydrogen, fluorine, chlorine, bromine, trifluoromethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH ₂ CH ₂ OCH ₃ ，-OCH ₂ CH(OH)CH ₂ OH, N-ethyl-amino or N-ethyl-N-methyl-amino;

in some embodiments, each R ₇ Independently selected from hydrogen or chlorine.

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

step 1:

step 2:

the following steps 3-1 or 3-2:

step 3-1:

step 3-2:

In some embodiments, when R ₆ When selected from hydrogen, the method comprises any one of step 3-1 or step 3-2.

In some embodiments, when R ₆ When selected from hydrogen, the method comprises a step 3-1; when R is ₆ Selected from optionally substituted alkyl groups, the process comprises the steps ofStep 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 、PdCl ₂ (PPh ₃ ) ₂ 、PdCl ₂ (PhCN) ₂ 、Pd(OAc) ₂ 、Pd/C、PdCl ₂ (dppf), or a combination thereof, and the like; in some embodiments, the catalyst in step 1 is PdCl ₂ (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 (C) _S F、K ₂ CO ₃ 、K ₃ PO ₄ Or a combination thereof, etc.; 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 solvents of the Suzuki reaction;

in some embodiments, step 1 is performed at room temperature or under heating conditions, e.g., 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, said step 3-2 is performed in a process selected from the group consisting of EtN (i-Pr) ₂ In the presence of a base.

In some embodiments, step 3-2 is performed in the presence of a solvent; in some embodiments, step 3-2 is performed 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 1a:

step 1a:

in some embodiments, wherein R ₀ ' is I, step 1a is performed in NIS and HBF ₄ Is carried out in the presence of (3). In some embodiments, R ₀ Is 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 CH ₃ CN, DMF, DMSO or THF, etc. can be miscible with water; in some embodiments, step 1a is performed at room temperature or under heating conditions, e.g., at a temperature of 80 ℃.

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

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

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

In some embodiments, the antineoplastic agent is selected from fluorouracil-based agents.

In some embodiments, the antineoplastic agent is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouridine, carmofur, doxifluridine, uracil tegafur, capecitabine, tegafur, cyprodinil, fluroxypyr, and Hilded.

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

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

in some embodiments, the tumor is a fluorouracil-resistant tumor.

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

In some embodiments, the fluorouracil is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouridine, carmofur, deoxyfluorouridine, uracil tegafur, capecitabine, tegafur, you-zidine, cyprodinil, fluroxypyr, and Hilded.

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

In some embodiments, the fluorouracil is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouridine, carmofur, deoxyfluorouridine, tegafur/uracil, capecitabine, tegafur, you-zidine, cyprodinil, fluroxypyr, and Hilded.

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

in the formula II:

Ar ¹ is optionally substituted non-aromatic heterocyclyl, optionally substituted aryl or optionally substituted heteroaryl;

Ar ² is optionally substituted non-aromatic heterocyclyl, or optionally substituted heteroaryl;

w is-NR ^a —、O、—CO—、—CONH—、—NHCH ₂ -or-CR ^b R ^c —；

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

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

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

Wherein R is ^a 、R ^b And R is ^c Each independently selected from H or optionally substituted alkyl.

In the formula III:

R ¹ represents (1) optionally substituted alkyl, (2) optionally substituted alkenyl, (3) optionally substituted alkynyl, (4) optionally substituted C _3-7 Carbocycles, or (5) -CONR ³¹ R ³² ；

R ³¹ And R is ³² Each independently represents (1) a hydrogen atom, or (2) an optionally substituted alkyl group;

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

R ³ and R is ⁴ Each independently represents (1) a halogen atom, (2) an optionally substituted alkyl group, (3) an optionally substituted alkoxy group, (4) a hydroxy group, (5) -L-CONR ⁶ R ⁷ ，(6)-L-SO ₂ R ⁸ Or (7) -L-COOR ⁹ ；

R ⁵ Represents (1) a hydrogen atom, (2) a halogen atom, (3) an optionally substituted alkyl group;

l represents (1) a 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,

[ 2 ]]

In the formula 2, A represents (1) a chemical bond, or (2) an oxygen atom; r is R ¹² And R is ¹³ Each independently represents (1) a hydrogen atom, (2) an optionally substituted alkyl group, (3) a hydroxyl group, or (4) NH ₂ Or (5) R ¹² And R is ¹³ Together with the carbon atom to which it is bound form C _3-7 A carbocycle; arrow on the right bonding-CONR ⁶ R ⁷ 、-SO ₂ R ⁸ or-COOR ⁹ ，

R ⁶ And R is ⁷ Each independently represents (1) a hydrogen atom, (2) an optionally substituted alkyl group, (3) a hydroxyl group, (4) -CONR ¹⁵ R ¹⁶ ，(5)-SO ₂ NR ¹⁵ R ¹⁶ ，(6)-COR ¹⁷ Or (7) -SO ₂ R ¹⁷ Or R is ⁶ And R is ⁷ Together with the nitrogen atom to which it is bonded, form a 4-7 membered nitrogen containing saturated heterocyclic ring optionally substituted with hydroxy;

R ⁸ represents (1) optionally substituted alkyl, or (2) NR ¹⁰ R ¹¹ ；

R ⁹ Represents (1) a hydrogen atom, or (2) an optionally substituted alkyl group;

R ¹⁰ and R is ¹¹ Each independently represents (1) a hydrogen atom, (2) an optionally substituted alkyl group, (3) -CONR ¹⁵ R ¹⁶ ，(4)-SO ₂ NR ¹⁵ R ¹⁶ ，(5)-COR ¹⁷ Or (6) -SO ₂ R ¹⁷ ；

E represents optionally substituted alkyl or

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

R ¹⁴ represents (1) a hydrogen atom, or (2) a hydroxyl group;

R ¹⁵ and R is ¹⁶ Each independently represents (1) a hydrogen atom, (2) an optionally substituted alkyl group, or (3) a 5-12 membered ring group;

R ¹⁷ represents (1) optionally substituted alkyl, or (2) a 5-12 membered ring group;

M ¹ and M ² Each independently represents (1) a bond, (2) -C (O) -, (3) -O-, -4) -S-, -5) -C (O) O-, -6) -CH ₂ O-, 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, a plurality of R ³ May be the same or different;

when q is 2 or more, a plurality of R ⁴ May be the same or different;

when R is 2 or more, a plurality of R ⁵ May be the same or different;

when t is 2 or more, a plurality of R ¹² And R is ¹³ May be the same or different, respectively.

In some embodiments, in the formula II:

Ar ¹ is an optionally substituted non-aromatic 5-18 membered heterocyclyl, optionally substituted C ₆ -C ₁₈ Aryl or optionally substituted C ₁ -C ₁₈ Heteroaryl;

Ar ² is optionally substituted non-aromatic 5-18 membered heterocyclyl, or optionally substituted C ₁ -C ₁₈ Heteroaryl;

w is-NR ^a —、O、—CO—、—CONH—、—NHCH ₂ -or-CR ^b R ^c —；

Z is-C (O) -C (S) -C,

/>

Y is-NR ^a -, O, or-CR ^b R ^c -a step; and

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

Wherein R is ^a 、R ^b And R is ^c Each independently selected from H or optionally substituted C _1-6 An alkyl group.

In some embodiments, in the formula III:

R ¹ represents (1) optionally substituted by 1 to 5R ²¹ Substituted C _1-8 Alkyl, (2) optionally substituted with 1 to 5R ²¹ Substituted C _2-8 Alkenyl, (3) optionally substituted with 1-5R ²¹ Substituted C _2-8 Alkynyl, (4) optionally being substituted with 1 to 5 substituents selected from C _1-4 Alkyl, C _1-4 Haloalkyl, C _1-4 C substituted by substituent groups of alkoxy groups and halogen atoms _3-7 Carbocycles, or (5) -CONR ³¹ R ³² ；

R ²¹ Represents (1) a halogen atom, (2) -OR ²² ，(3)-NR ²³ R ²⁴ Or (4) an oxo group, wherein,

R ²² represents (1) a hydrogen atom, (2) C _1-4 Alkyl or (3) C _1-4 A haloalkyl group, a halogen atom,

R ²³ and R is ²⁴ Each independently represents (1) a hydrogen atom, or (2) C _1-4 An alkyl group;

R ³¹ and R is ³² Each independently represents (1) a hydrogen atom, or (2) C _1-4 An alkyl group;

R ² represents (1) a hydrogen atom, (2) C _1-4 Alkyl, or (3) C _1-4 A haloalkyl group;

R ³ and R is ⁴ Each independently represents (1) a halogen atom, (2) C _1-4 Alkyl, (3) C _1-4 Haloalkyl, (4) C optionally substituted with halogen _1-4 Alkoxy, (5) hydroxy, (6) -L-CONR ⁶ R ⁷ ，(7)-L-SO ₂ R ⁸ Or (8) -L-COOR ⁹ ；

R ⁵ Represents (1) a hydrogen atom, (2) a halogen atom, (3) C _1-4 Alkyl, or (4) C _1-4 A haloalkyl group;

l represents (1) a bond, (2) a group represented by the following formula 2, (3) C _2-4 Alkenylene, (4) -O-C _2-4 Alkenylene, (5) an oxygen atom, or (6) optionally C _1-4 An alkyl-substituted nitrogen atom,

[ 2 ]]

In the formula 2, A represents (1) a chemical bond, or (2) an oxygen atom; r is R ¹² And R is ¹³ Each independently represents (1) a hydrogen atom, (2) C _1-4 Alkyl, (3) hydroxy, or (4) NH ₂ Or (5) R ¹² And R is ¹³ Together with the carbon atom to which it is bound form C _3-7 A carbocycle; arrow on the right bonding-CONR ⁶ R ⁷ 、-SO ₂ R ⁸ or-COOR ⁹ ，

R ⁶ And R is ⁷ Each independently represents (1) a hydrogen atom, (2) C _1-4 Alkyl, (3) C _1-4 Haloalkyl, (4) hydroxy, (5) -CONR ¹⁵ R ¹⁶ ，(6)-SO ₂ NR ¹⁵ R ¹⁶ ，(7)-COR ¹⁷ Or (8) -SO ₂ R ¹⁷ Or R is ⁶ And R is ⁷ Together with the nitrogen atom to which it is bonded, form a 4-7 membered nitrogen containing saturated heterocyclic ring optionally substituted with hydroxy;

R ⁸ representation (1) C _1-4 Alkyl, (2) C _1-4 Haloalkyl, or (3) NR ¹⁰ R ¹¹ ；

R ⁹ Represents (1) a hydrogen atom, or (2) C _1-8 An alkyl group;

R ¹⁰ and R is ¹¹ Each independently represents (1) a hydrogen atom, (2) C _1-4 Alkyl, (3) -CONR ¹⁵ R ¹⁶ ，(4)-SO ₂ NR ¹⁵ R ¹⁶ ，(5)-COR ¹⁷ Or (6) -SO ₂ R ¹⁷ ；

E represents alkyl optionally substituted by halogen or

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

R ¹⁴ represents (1) a hydrogen atom, or (2) a hydroxyl group;

R ¹⁵ and R is ¹⁶ Each independently represents (1) a hydrogen atom, (2) C _1-4 Alkyl, or (3) a 5-7 membered ring group;

R ¹⁷ representation (1) C _1-4 Alkyl, 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, a plurality of R ³ May be the same or different;

when q is 2 or more, a plurality of R ⁴ May be the same or different;

when R is 2 or more, a plurality of R ⁵ May be the same or different;

In some embodiments, in the formula II:

Ar ¹ is optionally substituted aryl or optionally substituted heteroaryl;

in some embodiments, ar ¹ Is 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; -SO ₂ -an alkyl group; -SO ₂ NR ^15a R ^15b ；-C(＝O)NR ^15c R ^15d The method comprises the steps of carrying out a first treatment on the surface of the Optionally substituted amino; a hydroxyl group; cyano group; and a carboxyl group; s represent 0, 1, 2, 3 or 4;

in some embodiments, ar ¹ Is aryl or heteroaryl, each independently optionally substituted with s groups independently selected from: halogen; -C _1-6 Alkyl optionally substituted with one or more independently selected R ¹³ Group substitution; -C _1-6 Alkoxy optionally substituted with one or more independently selected R ¹³ Group substitution; -C _3-7 Monocyclic cycloalkyl optionally substituted with one or more independently selected R ¹⁴ Group substitution; 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 R ¹⁴ Group substitution; -O-C _3-7 Monocyclic cycloalkyl optionally substituted with one or more independently selected R ¹⁴ Group substitution; -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 by one or more independently selected R ¹⁴ Group substitution; -SO ₂ -C _1-6 An alkyl group; -SO ₂ NR ^15a R ^15b ；-C(＝O)NR ^15c R ^15d ；-NR ^17a R ^17b The method comprises the steps of carrying out a first treatment on the surface of the A hydroxyl group; cyano group; and a carboxyl group;

wherein each R ¹³ Independently selected from: halogen; -CN; -OH; -C _1-4 Alkoxy optionally substituted with one or more independently selected OH, C _1-4 Alkoxy or halogen substitution; -C (=o) NR ^16a R ^16b ；

-NR ^16c C(＝O)-C _1-4 An alkyl group; -NR ^16d C(＝O)-C _1-4 An alkoxy group; -SO ₂ -C _1-4 An alkyl group; -SO ₂ NR ^16e R ^16f ；-NR ^16g SO ₂ -C _1-4 An alkyl group; -C _3-7 Monocyclic cycloalkyl optionally substituted C with one or more independently selected halogen or optionally substituted C with one or more halogen _1-4 Alkyl substitution; and 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 halogens or optionally substituted with one or more halogens _1-4 Alkyl substitution;

each R is ¹⁴ Independently selected from: halogen; -CN; -OH; -C _1-4 Alkoxy optionally substituted with one or more halogens; -C _1-4 Alkyl optionally substituted with one or more halogens;

each R is ^15a 、R ^15b 、R ^15c 、R ^15d 、R ^16a 、R ^16b 、R ^16c 、R ^16d 、R ^16e 、R ^16f And R is ^16g Independently selected from H and C _1-4 An alkyl group;

each R is ^17a And R is ^17b Independently selected from H and optionally one or more independently selected halogens, OH or C _1-4 Alkoxy substituted C _1-4 An alkyl group.

In some embodiments, ar ² Is Ar with ¹ The same monocyclic heteroaryl group is represented by either one of the following structures (1) to (3):

wherein in the structure (1):

R ₁ is hydrogen or optionally substituted alkyl;

R ₂ selected from hydrogen, deuterium, halogen, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted amino, optionally substituted cycloalkyl or optionally substituted cycloalkoxy;

R ₃ And R is ₄ Each independently selected from hydrogen, deuterium, halogen, -CN, -NO ₂ Nitroso, carboxyl, sulfonic, 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;

R ₃ and R is ₄ May be located at h, i or j, but not at the same location 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);

R ³ is optionally substituted alkyl;

x is selected from O, NR ⁴ And CR (CR) ⁴ R ⁵ Wherein R is ⁴ 、R ⁵ Each 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 ^1a selected from:

-C _1-4 alkyl optionally substituted with one or more groups independently selected from: OH; c (C) _1-4 Alkoxy, optionally substituted with one or more independently selected OH or C _1-4 Alkoxy substitution; -SO ₂ -C _1-4 An alkyl group; -O-C _3-7 A 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 ^6a R ^6b ；

-C _1-4 an alkoxy group;

-C _3-7 a monocyclic cycloalkyl group;

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 halo;

-O-C _3-7 monocyclic cycloalkyl

-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;

R ^1b is H or C _1-4 An alkyl group;

Cy ₁ is a 5-membered monocyclic heteroaryl ring containing one, two or three heteroatoms independently selected from N, O or S, or

Cy ₁ Is a 4 to 7 membered monocyclic heterocycloalkyl ring containing one, two or three heteroatoms independently selected from N, O or S, or a 4 to 7 membered monocyclic heterocycloalkyl ring containing one, two or three heteroatoms independently selected from N, O or S, fused to a 5 to 6 membered heteroaryl ring containing one, two or three heteroatoms independently selected from N, O or S, which heteroaryl can optionally be substituted with one C _1-4 Alkyl substitution;

R ³ selected from:

-C _1-4 alkyl optionally substituted with one or more independently selected of the following groups: halogen; -CN; -OH; -C _1-4 An alkoxy group; or-NR ^7a R ^7b ；

-C _1-4 Alkoxy substituted with one or more halogens;

-C _3-7 a monocyclic cycloalkyl group;

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

-CN；

-S(O) ₂ -C _1-4 an alkyl group;

-NR ^8a R ^8b the method comprises the steps of carrying out a first treatment on the surface of the And

-C(＝O)NR ^8c R ^8d ；

each R is ⁴ Independently selected from: -C _1-4 Alkyl optionally substituted with one or more independently selected R ¹² Group substitution; -C _3-7 A monocyclic cycloalkyl group; and 4 to 7 membered monocyclic heterocycloalkyl containing one, two or three heteroatoms independently selected from N, O and S;

each R is ¹² Independently selected from: -halogen; -OH; -C _1-4 An alkoxy group; -SO ₂ -C _1-4 An alkyl group; -C _3-7 Monocyclic cycloalkyl optionally substituted with one or more independently selected-OH, halogen, -CN, C _1-4 Alkyl, C _1-4 Alkoxy or = O substitution; 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-OH, halogen, -CN, C _1-4 Alkyl, C _1-4 Alkoxy or = O substitution; -NR ^9a R ^9b The method comprises the steps of carrying out a first treatment on the surface of the and-CN;

each R is ^6a 、R ^6b 、R ^7a 、R ^7b 、R ^8a 、R ^8b 、R ^8c 、R ^8d 、R ^9a 、R ^9b Independently selected from H and C _1-4 An alkyl group;

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

in some embodiments, when Ar ² In the case of the above-mentioned structure (3), X is-CR ^b R ^c -a step; y and W are-NR ^a -a step; z is-C (O) -or-C (=N-CN) -.

In some embodiments, in the formula III:

R ¹ Represents (1) optionally substituted by 1 to 5R ²¹ Substituted C _1-8 Alkyl, or (2) optionally substituted with 1 to 5 substituents selected from C _1-4 Alkyl, C _1-4 C substituted by substituent groups of alkoxy group, halogen atom and trifluoromethyl group _3-7 A carbocycle;

in some embodiments, R ¹ Is C _1-8 An alkyl group; in some embodiments, R ¹ Is C _1-6 An alkyl group; in some embodiments, R ¹ Is (CH) ₃ ) ₂ CHCH ₂ -or (CH) ₃ CH ₂ ) ₂ CH-；

In some embodiments, R ² Is a hydrogen atom;

in some embodiments, R ¹⁴ Is hydroxyl;

in some embodiments, M ¹ And M ² Each independently represents-O-or-S-;

in some embodiments, R ³ And R is ⁴ Each independently represents (1) a halogen atom, (2) C optionally substituted with halogen _1-4 Alkoxy, or (3) -L-SO ₂ R ⁸ ；

In some embodiments, L represents a bond;

in some embodiments, R ⁸ Represent C _1-4 An alkyl group;

in some embodiments, R ⁸ Is methyl;

in some embodiments, R ³ And R is ⁴ Each independently represents F, -SO ₂ CH ₃ or-OCH ₂ CH ₂ F；

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 aryl; in some embodiments, ring 1 and ring 2 each independently represent phenyl, cyclohexyl, pyridinyl; 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 the formula II:

R ^a 、R ^b and R is ^c Each independently selected from: hydrogen; or an alkyl group optionally substituted with a substituent independently selected from hydroxy, optionally substituted alkoxy, optionally substituted ester, optionally substituted aminocarbonyl, or carboxy;

in some embodiments, R ^a 、R ^b And R is ^c Each independently selected from: hydrogen; or optionally substituted C selected from hydroxy _1-4 C substituted by alkoxy, optionally substituted ester, optionally substituted aminocarbonyl, or carboxyl groups _1-6 An alkyl group;

in some embodiments, R ^a 、R ^b And R is ^c Each independently selected from: hydrogen; or C optionally substituted with a group selected from hydroxy, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl, or carboxyl _1-6 An alkyl group;

in some embodiments, R ^a 、R ^b And R is ^c Each independently selected from: hydrogen, methyl, ethyl, -CH ₂ COOH、-CH ₂ CH ₂ OH、-CH ₂ CH ₂ CH ₂ OH、-CH ₂ CONH ₂ Or CH (CH) ₂ COOEt；

In some embodiments, R ^a 、R ^b And R is ^c Each 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 ^a -a step; and

in some embodiments, X is a direct bond, -NR ^a -, or-CR ^b R ^c —；

In some embodiments of the present invention, in some embodiments,is-NH-NH-C (O) -NH-, -N (CH) ₃ ) -NH-C (O) -NH-, -NH-C (O) -NH-or-CH ₂ -NH-C(O)-NH-；

In some embodiments, at Ar ¹ Optionally substituted aryl groups appearing in the definition of (a) are optionally substituted C ₆ -C ₁₀ An aryl group;

in some embodiments, at Ar ¹ Optionally substituted heteroaryl groups appearing in the definition of (a) are optionally substituted monocyclic or fused bicyclic heteroaryl groups;

in some embodiments, at Ar ¹ Optionally substituted heteroaryl groups appearing in the definition of (a) are optionally substituted monocyclic 5-10 membered or 5-7 membered heteroaryl groups or optionally substituted fused bicyclic 8-12 membered heteroaryl groups;

in some embodiments, ar ¹ Is an optionally substituted phenyl or an optionally substituted 5-to 6-membered monocyclic heteroaryl comprising one or two heteroatoms independently selected from N, O and S;

in some embodiments, ar ¹ Is 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, ar ¹ Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; optionally by halogen, hydroxy or C _1-6 Alkoxy substituted C _1-6 An alkyl group; optionally by halogen, hydroxy or C _1-6 Alkoxy substituted C _1-6 An alkoxy group; or optionally by C _1-6 An alkyl-substituted amino group;

in some embodiments, ar ¹ Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; c optionally substituted by halogen, hydroxy _1-6 An alkyl group; optionally by hydroxy, C _1-6 Alkoxy substituted C _1-6 An alkoxy group; or optionally by C _1-6 An alkyl-substituted amino group;

in some embodiments, ar ¹ Is heteroaryl optionally substituted with s groups independently selected from: hydrogen; halogen; optionally is covered byHalogen substituted C _1-6 An alkyl group; optionally by hydroxy, C _1-6 Alkoxy substituted C _1-6 An alkoxy group;

in some embodiments, ar ¹ Is heteroaryl optionally substituted with s groups independently selected from: hydrogen, fluorine, chlorine, bromine, trifluoromethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH ₂ CH ₂ OCH ₃ ，-OCH ₂ CH(OH)CH ₂ OH, N-ethyl-amino or N-ethyl-N-methyl-amino;

in some embodiments, ar ¹ Is heteroaryl optionally substituted with s groups independently selected from: hydrogen, chlorine, bromine, trifluoromethyl, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH ₂ CH(OH)CH ₂ OH or-OCH ₂ CH ₂ OCH ₃ ；

In some embodiments, at Ar ¹ The heteroaryl groups present in the definition of (a) being optionally substituted with 1 to 2 of said substituents in the ortho position to the heteroatom;

in some embodiments, at Ar ¹ The heteroaryl groups appearing in the definition of (a) are pyridine or thiophene;

in some embodiments, ar ¹ Selected from 4-substituted 2, 6-dichloropyridine, 4-substituted 2, 6-dibromopyridine, 4-substituted 2-chloro-6-hydroxyethyl pyridine, 4-substituted-2-chloro-6-hydroxypropyl pyridine, 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, 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-dichloro-thiophene;

in some embodiments, ar ¹ Is 4-substituted 2, 6-dichloropyridine, 4-substituted-2-chloro-6- (2-methoxyethoxy)) Pyridine, 4-substituted-2-trifluoromethyl-6- (2-methoxyethoxy) pyridine, 4-substituted-2-trifluoromethyl-6-ethoxypyridine or 4-substituted-2-trifluoromethyl-6- (2, 3-dihydroxypropoxy) pyridine;

In some embodiments, ar ¹ And Ar is a group ² Are all 4-substituted 2, 6-dichloropyridine;

in some embodiments, ar ¹ Is 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, ar ¹ Is aryl optionally substituted with s groups independently selected from: hydrogen, halogen, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 Haloalkyl, C _1-6 Haloalkoxy, amino, mono-or di-C _1-6 Alkylamino, C _3-7 Cycloalkyl or C _3-7 A cycloalkoxy group.

In some embodiments, ar ¹ Is aryl optionally substituted with s groups independently selected from: hydrogen, halogen, C _1-6 Alkyl, C _1-4 Alkoxy, C _1-6 Perhaloalkyl, C _1-4 Perhaloalkoxy, amino, mono-or di-C _1-6 Alkylamino, C _3-7 Cycloalkyl or C _3-7 A cycloalkoxy group.

In the structure (1) of the formula II:

in some embodiments, R ₁ Is optionally substituted C _1-12 An alkyl group; in some embodiments, R ₁ Selected from C optionally substituted by halogen, alkenyl or hydroxy _1-6 An alkyl group; in some embodiments, R ₁ Selected from C optionally substituted by alkenyl or hydroxy _1-6 An alkyl group; in some embodiments, R ₁ Is methyl, ethyl, n-propyl, isopropyl, allyl or-CH ₂ CH ₂ CH ₂ OH; in some embodiments, R ₁ Is methyl, allyl or-CH ₂ CH ₂ CH ₂ OH; in some embodiments, R ₁ Is methyl;

in some embodiments, R ₂ Selected from hydrogen, deuterium, halogen, C _1-6 Alkyl, C _1-6 Alkoxy, C _1-6 Perhaloalkyl, C _1-6 Perhaloalkoxy, amino, mono-or di-C _1-6 Alkylamino, C _3-7 Cycloalkyl or C _3-7 A cycloalkoxy group; in some embodiments, R ₂ Selected from hydrogen and C _1-6 An alkyl group; in some embodiments, R ₂ Selected from hydrogen and methyl;

in some embodiments, R ₃ And R is ₄ Each independently selected from hydrogen, deuterium, halogen, -CN, -NO ₂ Nitroso, carboxyl, sulfonic acid, optionally substituted sulfonyl, optionally substituted C _1-6 Alkyl, optionally substituted C _1-6 Alkoxy, optionally substituted amino, optionally substituted C _3-7 Cycloalkyl, optionally substituted C _3-7 Cycloalkoxy, optionally substituted C ₆ -C ₁₈ Aryl or optionally substituted C ₁ -C ₁₈ Heteroaryl;

in some embodiments, R ₃ And R is ₄ Each independently selected from: hydrogen; and C independently of each other optionally substituted with _1-6 Alkyl, C _3-7 Cycloalkyl, C ₆ -C ₁₈ Aryl or C ₁ -C ₁₈ Heteroaryl group: halogen, C ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl or C _1-6 An alkoxy group;

in some embodiments, R ₃ And R is ₄ Each independently selected from: hydrogen; c (C) _3-7 Cycloalkyl; optionally by halogen, C ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl or C _1-6 Alkoxy substituted C _1-6 An alkyl group; each independently optionally substituted with halogen, or C _1-6 Alkoxy substituted C ₆ -C ₁₈ Aryl or C ₁ -C ₁₈ Heteroaryl;

in some embodiments, R ₃ And R is ₄ Each independently selected from: hydrogen; c (C) _3-7 Cycloalkyl; optionally by halogen or C ₆ -C ₁₈ Aryl substituted C _1-6 An alkyl group; optionally by halogen, or C _1-6 Alkoxy substituted C ₆ -C ₁₈ An aryl group; or C optionally substituted by halogen ₁ -C ₁₈ Heteroaryl;

in some embodiments, R ₃ And R is ₄ Each independently is independently selected from: hydrogen; c (C) _3-6 Cycloalkyl; c optionally substituted by halogen or phenyl _1-6 An alkyl group; optionally by halogen, or C _1-6 Alkoxy substituted phenyl; or furyl or thienyl each independently optionally substituted with halogen;

in some embodiments, R ₃ And R is ₄ Each independently selected from: hydrogen, methyl, n-propyl, isopropyl, cyclopropyl, cyclohexyl, -CF ₃ Benzyl, phenyl, fluoro-substituted phenyl, methoxy-substituted phenyl, furyl or chloro-substituted thienyl;

in some embodiments, R ₃ And R is ₄ Is 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), R ¹ And R is ² Independently selected from H and alkyl, methoxy, hydroxy, halogen, nitrile, trifluoromethyl and carboxy;

R ³ independently selected from alkyl, methoxy, hydroxy, halogen, nitrile, trifluoromethyl and carboxy;

d is CR or N;

x is O, NR ⁴ ，CR ⁴ R ⁵ Wherein R occurs in the definition of X ⁴ And R is ⁵ Independently selected from: h and alkyl, which is, for example, lower alkyl and may have 1 to 10 carbons, and may be cyclic or branched alkyl having 3 to 10 carbons; methoxy; a hydroxyl group;f, performing the process; br; i, a step of I; a nitrile; trifluoromethyl and carboxyl;

y is O or S;

z is a substituted aromatic ring having the following structure:

wherein R is ₆ And R is ₇ Independently selected from: alkyl and may comprise 1 to 10 carbons, and may be a cyclic or branched alkyl having 3 to 10 carbons; methoxy; 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), R ¹ And R is ² Independently selected from H and alkyl, methoxy, hydroxy, halogen, nitrile, trifluoromethyl and carboxy;

x is O, NR ⁴ ，CR ⁴ R ⁵ Wherein R is ⁴ And R is ⁵ Independently selected from: h and alkyl, which is, for example, lower alkyl and may have 1 to 10 carbons, and may be cyclic or branched alkyl having 3 to 10 carbons; methoxy; a hydroxyl group; f, performing the process; br; i, a step of I; a nitrile; trifluoromethyl and carboxyl;

y is O or S;

z is a substituted aromatic ring having the following structure:

wherein R is ₆ And R is ₇ Independently selected from: alkyl group toAnd may comprise 1 to 10 carbons, and may be a cyclic or branched alkyl group having 3 to 10 carbons; methoxy; a hydroxyl group; halogen; a nitrile; trifluoromethyl and carboxyl; and

e is N or CR.

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

in some embodiments, R ^1a Selected from methyl;

in some embodiments, R ^1b Selected from hydrogen;

in some embodiments of the present invention, in some embodiments,is->

In some embodiments, R ³ And R is ⁴ Each independently selected from hydrogen or C optionally substituted with halogen _1-6 An alkyl group;

in some embodiments, R ³ Is methyl;

in some embodiments, R ⁴ Is hydrogen or trifluoromethyl; in some embodiments, R ⁴ At least one of which is trifluoromethyl.

In some embodiments, the S1PR2 inhibitor is a compound or a pharmaceutically acceptable salt, or tautomer, or pharmaceutical composition thereof, or a combination of same 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, gastric 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 an anti-drug resistant tumor therapeutic agent, wherein the anti-drug resistant tumor therapeutic agent is a tumor therapeutic agent capable of resisting and/or reversing the drug resistance of fluorouracil-based agents, the method comprising:

providing a cell line or cell culture expressing S1PR 2;

contacting a candidate object 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 the expression level of S1PR2 with a control cancer cell line or cancer cell culture not contacted with the candidate;

when the expression level of S1PR2 is lower than the control cancer cell line or cancer cell culture, the candidate is indicated to be able to act as an anti-drug resistant tumor therapeutic.

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

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

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

detecting the level of expression of DPD in said cell line or cell culture after contact with the candidate and comparing the level of expression of DPD with 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, the candidate is indicated to be capable of acting as an anti-drug resistant tumor therapeutic;

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

optionally an antitumor drug.

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

Detailed Description

The following describes the embodiments of the present invention in further detail with reference to examples. The following examples are for illustrative purposes only and are not intended to limit the scope of the present application.

Preparation example 1

The synthetic methods or routes reported in references or patents (including but not limited to, for example, those mentioned in the background section above) produce the following known S1PR2 inhibitors:

preparation example 2:

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

the synthetic route I comprises the following steps:

step 1a:

step 1:

step 2:

the following steps 3-1 or 3-2:

step 3-1:

step 3-2:

/>

synthesis of Compound 9-1:

1 methyl-5-aminopyrazole (1) and 2, 6-dichloropyridine-4 carboxylic acid are used as initial raw materials, and the preparation method comprises the following steps: namely, 1 methyl-5-aminopyrazole (1, 30.93 mmol) and ethyl isobutyrylacetate (37.11 mmol) were refluxed at 140℃for 20 hours with acetic acid as a solvent, cooled to room temperature, dried with spin-drying the solvent, added with 10mL of ethyl acetate, stirred for ten minutes, and then suction-filtered to obtain white solid 1.

Compound 2 (1.95 mmol) and POBr ₃ (2.54 mmol) anisole as solvent, refluxing at 120deg.C for 3h, cooling to room temperature, washing with saturated aqueous sodium chloride solution, extracting with dichloromethane, combining organic layers, and concentrating with anhydrous MgSO ₄ Drying, spin-drying the solvent, and column chromatography to obtain white solid 3.

Compound 3 (200 mg,0.89 mmol), NIS (200 mg,0.89 mmol), 48% HBF ₄ (399mg, 4.44 mmol) in CH ₃ CN (5 mL), reflux at 80℃for 2h. TLC detection of completion of the reaction, pouring the reaction solution into 1M NaHCO ₃ The aqueous solution was filtered and washed with water to give compound 4 (237 mg, yield 76%). ¹ H NMR(600MHz,DMSO-d ₆ )δ7.13(s,1H),3.90(s,3H),2.62(s,3H). ¹³ C NMR(125MHz,DMSO-d ₆ )δ149.51,145.89,141.64,122.09,116.71,89.24,34.61,17.33.

Compound 4 (1.0 g,2.85 mmol), cs ₂ CO ₃ (1.9g,5.7mmol),PdCl ₂ (PPh ₃ ) ₂ (100 mg,0.14 mmol), and 4-pyridineboronic acid (350 mg,2.85 mmol) in 1, 4-dioxane/H ₂ O (15 mL, v/v 3/1). The reaction system was purged of air, purged with nitrogen and stirred at 80℃for 12 hours. TLC detection was complete, the reaction mixture was washed with water and saturated aqueous NaCl, extracted with dichloromethane, and anhydrous MgSO ₄ Drying, filtering, spin-drying the solvent, and column chromatography gave compound 5-1 (410 mg, 52%). ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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.

Compound 5-1 (1.80 mmol) and 80% hydrazine hydrate (89.89 mmol) were refluxed for 10h at 100 ℃ with ethanol as solvent, cooled to room temperature, the solvent was dried by spinning, washed with saturated aqueous sodium chloride solution, extracted with dichloromethane, the organic layers were combined, and dried over anhydrous MgSO ₄ Drying and spin-drying the solvent to obtain white solid 6-1.

2, 6-dichloropyridine-4-carboxylic acid (26 mmol) was placed in a double-necked flask, 30mL of 1, 4-dioxane was added under nitrogen protection, triethylamine (39 mmol) was added dropwise, diphenyl azide phosphate DPPA (39 mmol) was added dropwise under ice bath, the reaction was allowed to proceed overnight, TLC detection was complete, washing with saturated aqueous sodium chloride solution, dichloromethane extraction, combining the organic layers, and using anhydrous MgSO ₄ Drying, spin-drying the solvent, and column chromatography to obtain white solid 8.

Dissolving compound 8 (1.68 mmol) in 3mL toluene, heating at 80deg.C for 3h, TLC detecting complete conversion to isocyanate, cooling to 50deg.C, adding tetrahydrofuran solution of compound 6-1 (1.68 mmol), reacting for 5h, TLC detecting complete reaction, cooling to room temperature, spin-drying solvent, washing with saturated aqueous sodium chloride solution, extracting with dichloromethane, combining organic layers, and concentrating with anhydrous MgSO ₄ Drying, spin drying solvent, and column chromatography to obtain white solid 9-1.

9-1： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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 can be prepared by reference to the above synthetic routes.

9-2： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )δ151.98,149.84,142.38,139.44,132.54,130.67,119.25,110.97,33.90,20.65.

9-3： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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： ¹ H NMR(400MHz,DMSO-d ₆ )δ9.97(s,2H),7.55(s,4H). ¹³ C NMR(100MHz,DMSO-d ₆ )δ151.63,150.84,150.12,111.93.

16-6： ¹ H NMR(500MHz,DMSO-d ₆ )δ9.80(s,1H),8.99(s,2H),8.07–7.34(m,8H),6.74(s,1H),3.91(s,3H). ¹³ C NMR(125MHz,DMSO-d ₆ )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: ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )δ159.85,153.42,152.01,150.34,149.83,131.22,111.70,100.38,33.70,32.03,22.86.

16-8: ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )δ159.80,151.99,150.13,149.82,130.90,111.69,109.59,98.47,33.70,14.18,10.51.

16-9: ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )δ159.82,152.00,150.00,149.83,131.29,111.75,103.65,55.35,33.74,18.62.

16-10: ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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: ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )δ159.74,151.99,150.98,149.81,143.99,140.16,111.69,103.26,33.21,19.23,14.91.

16-12: ¹ H NMR(500MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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: ¹ H NMR(500MHz,DMSO-d ₆ )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: ¹ H NMR(600MHz,DMSO-d ₆ )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: ¹ H NMR(600MHz,DMSO-d ₆ )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). ¹³ C NMR(125MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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: ¹ H NMR(400MHz,DMSO-d ₆ )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). ¹³ C NMR(100MHz,DMSO-d ₆ )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 test of S1PR2 inhibitor

In vitro cell activity experiments were performed on the known compounds listed in preparation example 1 above and the compounds synthesized in preparation example 2 and preparation example 3 of the present invention.

In vitro cell activity assay: DPD expression level verification by pre-pharmacodynamic activity screening and Western Blotting (Western Blotting) was performed in a variety of colon cancer and normal colon cell lines, and HCT116 and NCM460 were determined to be significantly 5-FU resistant cell lines. The log phase grown HCT116 and NCM460 cells were collected, the cell suspension concentration was adjusted, 100. Mu.L was added to each well, and the cells to be tested were plated to a density of 1000-10000 wells (the wells at the edges were filled with sterile PBS). 5% CO ₂ Incubation at 37 ℃ was carried out until the cell monolayer was confluent with the bottom of the wells (96 well flat bottom plate), and a concentration gradient of S1PR2 inhibitor compound and a constant concentration of 5-FU was added. 6 concentration gradients were set, 100 μl per well, for a total of 3 multiplex wells. 5% CO ₂ Incubate at 37℃for 24h and observe under an inverted microscope. mu.L of MTT solution (5 mg/mL, i.e., 0.5% MTT) was added to each well and the incubation was continued for 4 hours. The culture was terminated and the in-well culture solution was carefully aspirated. 150 μl of dimethyl sulfoxide was added to each well, and the mixture was shaken on a shaker at low speed for 0.5min to dissolve the crystals thoroughly. Absorbance was measured for each well at OD 560nm in an enzyme-linked immunosorbent assay. Cell viability is expressed as T/C%, T is the OD of the dosed cells, and C is the OD of the control cells. Calculation of EC from cell viability at each concentration gradient ₅₀ And IC ₅₀ The results are shown in Table 1.

TABLE 1

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Note that: EC (EC) ₅₀ EC for the combination of S1PR2 inhibitor with 5-FU ₅₀ Values, wherein the 5-FU concentration was 20. Mu.M. IC (integrated circuit) ₅₀ Is a compoundIC for single use ₅₀ Values, 5-FU in the absence of S1PR2 inhibitor, DMSO in the solvent, HCT116 and NCM460 in the presence of 5-FU resistant cells, indicate no cell death or undetectable.

TABLE 2

Note that: EC (EC) ₅₀ EC for the combination of S1PR2 inhibitor with 5-FU ₅₀ Values, wherein the 5-FU concentration was 20. Mu.M. IC (integrated circuit) ₅₀ IC for compound alone ₅₀ Values, 5-FU, in the absence of S1PR2 inhibitor, DMSO in the solvent, SW620 in 5-FU resistant cells, indicate no cell death or undetectable.

The results show that the 5-FU drug resistance of two cells under the action of 20 mu M5-fluorouracil, the S1PR2 inhibitor and the compound can reverse the drug resistance of the 5-fluorouracil, and the proliferation of the drug-resistant cells can be effectively inhibited by combining with the 5-fluorouracil.

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

The HCT116 cells (solvent control group, 10. Mu.M administration group to which the compounds of Table 1 were administered alone) and SW620 cells (solvent control group, 10. Mu.M administration group to which the compounds of Table 2 were administered alone (wherein 16-5 compounds were selected from 0.6. Mu.M administration group)) were removed, respectively, followed by cell scraping, protein extraction and quantification.

Cell protein extraction: firstly, PBS (phosphate buffered saline) is used for cleaning cells, each group of cells is placed at 4 ℃ for precooling, and cell lysate is prepared: NP-40 (960. Mu.L) +PPI (40. Mu.L) +PMSF (40. Mu.L). PBS was discarded, 100-200. Mu.L of lysate was added depending on the cell density, and the mixture was placed at 4℃for 40min. The cells were scraped off with a cell scraper and collected in a 1.5mL centrifuge tube and centrifuged at 12000rpm for 15 min. The supernatant was collected in another 1.5mL centrifuge tube, and in a 96-well plate, 90. Mu.L of the G250 staining solution was added, and 10. Mu.L of the protein standard solution and 10. Mu.L of the sample were added, respectively, to prepare a multiplex well. The concentration of cells is determined by an enzyme-labeled instrument, the concentration of cells in each group is regulated to be consistent, and 5x loading buffer is added to ensure that the concentration is 1x, and the metal bath is boiled for 5min.

Western blot detection: preparing separating gel and concentrated gel, and soaking in electrophoresis liquid. An equal amount of 20 mug protein sample is added into the loading well, and a marker or 1x loading buffer is added for edge sealing. And (3) performing constant-pressure electrophoresis at 80V until the protein is reduced to be separated from the gel, changing the gel into 120V, and continuing the constant-pressure electrophoresis. A PVDF membrane was used for 100V,90min transfer. The PVDF membrane was washed on a shaker for 5min once for 2 times. The blocking solution blocked the PVDF membrane for 1h and incubated with DPD primary antibody overnight at 4 ℃. The PVDF membrane was washed with 1xTBST on a shaker for 10min once for 3 times. The secondary antibody was incubated for 40min. The PVDF membrane was washed with 1xTBST on a shaker for 10min once for 3 times. ECL luminophores were formulated, uniformly sprayed onto PVDF film, chemiDoc XRS and molecular imager (molecular imager) exposure. Calculating a Bio-Rad quality One analysis mark gray scale ratio, and calculating a formula: protein of interest relative expression level= (protein of interest gray value-background gray value)/(internal reference protein gray value-background gray value).

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

Example 3: in vivo reverse 5-fluorouracil resistance experiment

The 5-fluorouracil and S1PR2 inhibitors were administered in combination using a nude mice subcutaneous inoculation tumor model.

BALB/c nude mice were purchased 6-8 weeks old, 6 each, and kept in a clean (SPF) environment without specific pathogens. Subcutaneously inoculating the 5-FU drug-resistant HCT116 cells, increasing tumor blocks to a certain size after two weeks, carrying out aseptic separation of tumor blocks, uniformly cutting, inoculating to armpits of each nude mouse, feeding in an aseptic environment for two weeks, and starting administration after the volume of each group of tumor blocks is consistent. The tail intravenous injection of 20mg/kg 5-fluorouracil combined with 25mg/kg JTE-013 and 25mg/kg 16-6 is continued for three weeks and half daily, compared with the single 5-fluorouracil group and the solvent group, wherein the solvent volume ratio is 2%DMSO+30%PEG 300+5%Tween 80+ddH ₂ O, the body weight and tumor volume changes (measured every 4 days) of nude mice were recorded throughout. Mice were then sacrificed and tumor samples and other tissues were removed for relevant detection and analysis of protein expression.

The results show that the nude mice tumor inoculated with the HCT116 cell strain has obvious drug resistance phenomenon to 5-fluorouracil treatment, the inhibition rate of 5-fluorouracil in vivo is only 13.01%, and the inhibition rate of the compound 16-6 can be improved to 46.52%. Whereas JTE-013 significantly increased the therapeutic sensitivity of tumor cells to 5-fluorouracil, increasing its inhibition to 75.87%, it was seen that S1PR2 inhibitors such as JTE-013 and compound 16-6 were effective in reversing the tumor cell resistance to 5-fluorouracil (see FIG. 2, where NC is a solvent group without 5-FU and compound, but with DMSO and water).

Example 4: in vivo inhibition of DPD expression

Western blot was performed on the difference in DPD expression in tumor tissue, normal colon and liver tissue of each group of nude mice.

Taking out each group of nude mice tumor tissue, normal colon and liver tissue respectively, 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.

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

Taking part of S1PR2 inhibitors as examples in the above in vitro experiments and JTE-013 (compound 14) and compound 16-6 as examples in the in vivo experiments, it was verified that S1PR2 inhibitors can effectively inhibit DPD expression, reverse 5-fluorouracil resistance, and can effectively inhibit tumor growth in combination with 5-fluorouracil. The present invention also verifies that other inhibitors of S1PR2 have similar effects in the same manner and the experimental procedure will not be described in detail here.

Example 5: knocking down the condition of in vitro inhibition of DPD expression of S1PR2

The colorectal cancer cell lines HCT116, SW620, liver cancer cell Huh7 and normal liver cell cccHEL-1 were subjected to shS PR2 lentivirus knockdown, respectively, and the DPD expression differences of the cell lines were subjected to western blot detection.

Short hairpin RNA lentivirus construction and packaging of S1PR2 was provided by beijing carrier, with a S1PR2 knockdown sequence of AGGAACAGCAAG TTCTACTCA. Cells were 4X 10 per well 24h before infection ⁴ Inoculating into 6-well plate, culturing in RPMI-1640 medium containing 10% FBS, culturing until cells reach 50% density after adhering to wall, adding filtered virus solution into 6-well plate, adding into 5% CO ₂ After virus infection for 48 hours, the infected cells were observed by a fluorescence microscope and were screened for green fluorescence using 10% FBS RPMI-1640 medium supplemented with puromycin to obtain a stable knockdown cell line. And performing western blot detection to obtain the expression difference of S1PR2 and DPD of each cell line.

The results show that the four cell lines HCT116, SW620, huh7 and cccHEL-1 all have significantly reduced S1PR2 compared with the empty vector group, and have higher knockdown efficiency. Meanwhile, the DPD expression levels of the four cell lines are also obviously down-regulated (see fig. 4), which proves that the knocking-down of the S1PR2 can effectively reduce the DPD expression, and further verifies that the S1PR2 inhibitor can effectively reduce the DPD expression.

In conclusion, the invention can regulate or inhibit the DPD expression level of cancer cells by targeted inhibition or regulation of the S1PR2 expression level, and can prevent the DPD expression level from degrading into the 5-fluorouracil of the cancer cells, thereby realizing the effect of reversing the drug resistance of the 5-FU. Since other 5-fluorouracil derivatives can be degraded into 5-FU to enter blood and finally enter cancer cells, inhibitors that inhibit DPD production by cancer cells by targeted inhibition or modulation of S1PR2 expression are suitable for drug-resistant treatment regimens for fluorouracil-like drug-treated cancers.

While the invention has been described in detail with respect to the general description and specific embodiments thereof, it will be apparent to those skilled in the art that various modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and 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:

R ₁ selected from each independently optionally substituted with halogen, -OH, -CN, C _1-6 Alkyl, C _1-6 Alkoxy or C ₆ -C ₁₈ Aryloxy substituted C ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl, or 5-10 membered heterocyclenyl;

R ₂ Selected from hydrogen or C _1-6 An alkyl group;

each R is ₃ Independently selected from: hydrogen and C _1-6 An alkyl group;

R ₄ 、R ₅ and R is ₆ Each independently selected from: hydrogen or C _1-6 An alkyl group;

ring B is a pyridine ring;

each R is ₇ Independently selected from hydrogen and halogen;

x is an integer from 1 to 2; and

y is an integer of 1, 2 or 3;

wherein said C ₁ -C ₁₈ Heteroaryl and the 5-10 membered heterocycloalkenyl contain 1 to 5 heteroatoms selected from N, O and S as ring-forming atoms.

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

R ₁ selected from each independently optionally substituted with halogen, -OH, -CN, C _1-6 Alkyl, C _1-6 Alkoxy or C ₆ -C ₁₈ Aryloxy substituted C ₆ -C ₁₂ Aryl, C ₁ -C ₁₂ Heteroaryl, or 5-10 membered heterocyclenyl.

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

R ₁ selected from C independently optionally substituted with fluorine, chlorine, -OH, -CN, methyl, ethyl, n-propyl, isopropyl, n-propoxy, isopropoxy or phenoxy ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl, or 5-10 membered heterocyclenyl.

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

R ₁ selected from C independently optionally substituted with fluoro, -OH, -CN, methyl, n-propoxy, isopropoxy or phenoxy ₆ -C ₁₈ Aryl, C ₁ -C ₁₈ Heteroaryl, or 5-10 membered heterocyclenyl.

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

R ₁ selected from: optionally selected from halogen, -OH, -CN, C _1-6 Alkyl, C _1-6 Alkoxy, or C ₆ -C ₁₈ Group-substituted C of aryloxy group ₆ -C ₁₈ An aryl group; optionally by C _1-6 Alkoxy substituted C ₁ -C ₁₈ Heteroaryl; or a 5-to 10-membered heterocycloalkenyl group.

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

R ₁ the C occurring in the definition of (2) ₆ -C ₁₈ Aryl is phenyl.

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

R ₁ the C occurring in the definition of (2) ₁ -C ₁₈ Heteroaryl is selected from pyridinyl or furanyl.

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

the 5-10 membered heterocycloalkenyl is

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

the halogen is selected from fluorine or chlorine.

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

R ₁ Selected from: phenyl optionally substituted with a group selected from fluoro, -OH, -CN, methyl, n-propoxy, or phenoxy; isopropoxy substituted pyridinyl; a furyl group; or alternatively

11. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R ₂ selected from C _1-6 An alkyl group.

12. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R ₂ selected from C _1-4 An alkyl group.

13. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R ₂ is methyl, ethyl, n-propyl or isopropyl.

14. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R ₂ is methyl.

15. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

each R is ₃ Independently selected from C _1-6 An alkyl group.

16. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

Each R is ₃ Independently selected from C _1-4 An alkyl group.

17. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

each R is ₃ Independently selected from hydrogen, methyl, n-propyl, isopropyl.

18. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

each R is ₃ Independently selected from hydrogen or methyl.

19. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

at least one R ₃ Is not hydrogen and is located para to N.

20. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

x is 1.

21. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R ₄ 、R ₅ and R is ₆ Each independently selected from: hydrogen; or C _1-4 An alkyl group.

22. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R ₄ 、R ₅ and R is ₆ Each independently selected from: hydrogen, methyl or ethyl.

23. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R ₄ 、R ₅ and R is ₆ Each independently selected from hydrogen or methyl.

24. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein:

R ₄ 、R ₅ and R is ₆ Are all hydrogen.

25. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein

The ortho position to the heteroatom in ring B is optionally substituted with 1-2R ₇ And (3) group substitution.

26. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein

Selected from pyridyl optionally substituted with: fluorine, chlorine or bromine.

27. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein

Selected from 4-substituted 2, 6-dichloropyridine or 4-substituted 2, 6-dibromopyridine.

28. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein

Is 4-substituted 2, 6-dichloropyridine.

29. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein

Each R is ₇ Independently selected from hydrogen, fluorine, chlorine or bromine.

30. The compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt, or tautomer thereof, wherein

Each R is ₇ Independently selected from hydrogen or chlorine.

31. The following compounds, or pharmaceutically acceptable salts, or tautomers thereof:

32. a process for the preparation of a compound as claimed in claim 1, comprising:

step 1:

step 2:

the following steps 3-1 or 3-2:

step 3-1:

step 3-2:

wherein R is ₀ And R is ₀ ' each independently is halogen; ring B, R ₁ 、R ₂ 、R ₃ 、R ₄ 、R ₅ 、R ₆ 、R ₇ X, y are as defined in claim 1.

33. The method of claim 32, wherein

When R is ₆ When selected from hydrogen, the method comprises any one of step 3-1 or step 3-2.

34. The method of claim 32, wherein

When R is ₆ When selected from hydrogen, the method comprises a step 3-1; when R is ₆ Selected from C _1-6 In the case of alkyl, the method includes step 3-2.

35. The method of claim 32, wherein

Step 1 is carried out in the presence of a catalyst.

36. The method of claim 32, wherein

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

37. The method of claim 36, wherein

The catalyst in the step 1 is PdCl ₂ (dppf)。

38. The method of claim 32, wherein

Step 1 is carried out in the presence of a base.

39. The method of claim 32, wherein

Step 1 is carried out in the presence of a base selected from the group consisting of: c (C) _S F、K ₂ CO ₃ 、K ₃ PO ₄ Or a combination thereof.

40. The method of claim 32, wherein

Step 3-2 is performed in the presence of a base.

41. The method of claim 32, wherein

Said step 3-2 is performed in a process selected from EtN (i-Pr) ₂ In the presence of a base.

42. The method of claim 32, wherein

Step 3-2 is performed in the presence of a solvent; optionally, said step 3-2 is performed in the presence of a solvent selected from DCM.

43. The method of claim 32, wherein

The method further comprises the following step 1a:

step 1a:

wherein R is ₀ ' is I, step 1a is performed in NIS and HBF ₄ Is carried out in the presence of (3).

44. A pharmaceutical composition comprising a compound of any one of claims 1-31, a pharmaceutically acceptable salt, or tautomer thereof.

45. The pharmaceutical composition of claim 44, wherein

The pharmaceutical composition also comprises pharmaceutically acceptable auxiliary materials.

46. The pharmaceutical composition of claim 44, wherein

The pharmaceutical composition further comprises one or more additional therapeutic agents.

47. The pharmaceutical composition of claim 46, wherein

The other therapeutic agent is an anti-tumor drug.

48. The pharmaceutical composition of claim 47, wherein

The antitumor drug is selected from fluorouracil drugs.

49. The pharmaceutical composition of claim 47, wherein

The antitumor drug is selected from the group consisting of 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouridine, carmofur, deoxyfluorouridine, uracil tegafur, capecitabine, tegafur, eufudine, cyprohexadine, fluroxypyr and Hilded.

50. Use of a compound of any one of claims 1-31, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of any one of claims 44-49, or a combination of same and 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 benefiting from inhibition of DPD expression; or in the preparation of a medicament for resisting and/or reversing the resistance of fluorouracil medicaments.

Use of an s1pr2 inhibitor, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition thereof, or a combination of same 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 benefiting from inhibition of DPD expression; or in the preparation of a medicament for the resistance and/or reversal of fluorouracil-based drug resistance;

Wherein:

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

in the formula II:

Ar ¹ is pyridinyl optionally substituted with s groups independently selected from: halogen; c optionally substituted by halogen _1-6 An alkyl group; optionally by hydroxy, or C _1-6 Alkoxy substituted C _1-6 An alkoxy group;

s represent 0, 1, 2, 3 or 4;

Ar ² is with Ar ¹ The same pyridyl group is represented by either the following structure (1) or structure (3):

w is-NR ^a —；

Z is-C (O) -,

y is-NR ^a -a step; and

x is a direct bond, -NR ^a -, or-CR ^b R ^c -a step; wherein R is ^a 、R ^b And R is ^c Each independently selected from H or C _1-6 An alkyl group;

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

R ₁ is C _1-6 An alkyl group;

R ₂ selected from hydrogen or C _1-6 An alkyl group;

R ₃ and R is ₄ Each being located at h, i or j, but not at the same location at the same time;

R ₃ and R is ₄ Is not hydrogen and is located at h; and said group located on h is selected from: c optionally substituted by halogen _1-6 An alkyl group; optionally by halogen or C _1-6 Alkoxy substituted C ₆ -C ₁₈ An aryl group;

the group located on i or j is selected from: hydrogen; optionally by C ₆ -C ₁₈ Aryl substituted C _1-6 An alkyl group;

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

R ^1a selected from C _1-4 An alkyl group;

R ^1b selected from hydrogen;

is->

R ³ And R is ⁴ Each independently selected from hydrogen or C optionally substituted with halogen _1-6 An alkyl group;

in the formula III:

R ¹ is C _1-8 An alkyl group;

R ² is a hydrogen atom or C _1-4 An alkyl group;

R ¹⁴ is hydroxyl;

R ⁵ represents a hydrogen atom;

e represents C optionally substituted by halogen _1-6 Alkyl or

Ring 1 and ring 2 each independently represent a 6-12 membered aryl group;

M ¹ and M ² Each independently represents-O-or-S-;

R ³ and R is ⁴ Each independently represents (1) a halogen atom, (2) C optionally substituted with halogen _1-4 Alkoxy, or (3) -L-SO ₂ R ⁸ ；

L represents a chemical bond;

R ⁸ represent C _1-4 An alkyl 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.

52. The use according to claim 50 or 51, wherein

The disease, disorder or condition that benefits from inhibition of DPD expression is selected from tumors.

53. The use according to claim 50 or 51, wherein

The disease, disorder or condition or the tumor is selected from the group consisting of a tumor of the digestive tract, a tumor of the head and neck, lung cancer, breast cancer.

54. The use according to claim 53, wherein

The head and neck tumor is selected from oral cancer.

55. The use according to claim 50 or 51, wherein

The tumor is a tumor with drug resistance to fluorouracil drugs.

56. The use according to claim 50 or 51, wherein

The one or more other therapeutic agents are anti-tumor agents.

57. The use according to claim 50 or 51, wherein

The antitumor drug is selected from fluorouracil drugs.

58. The use according to claim 50 or 51, wherein

The fluorouracil medicine is selected from 5-fluorouracil, 5'-2' -deoxyuridine, tegafur, fluorouridine, carmofur, deoxyfluorouridine, uracil tegafur, capecitabine, tegafur, eufutidine, cyprodinil, flurbiprofen and Hilded.

59. The method according to claim 51,

wherein, in the formula III:

R ¹ is C _1-6 An alkyl group.

60. The method according to claim 51,

wherein, in the formula III:

R ¹ is (CH) ₃ ) ₂ CHCH ₂ -or (CH) ₃ CH ₂ ) ₂ CH-。

61. The method according to claim 51,

wherein, in the formula III:

R ² is a hydrogen atom.

62. The method according to claim 51,

wherein, in the formula III:

R ⁸ is methyl.

63. The method according to claim 51,

wherein, in the formula III:

R ³ and R is ⁴ Each independently represents F, -SO ₂ CH ₃ or-OCH ₂ CH ₂ F。

64. The method according to claim 51,

wherein, in the formula III:

ring 1 and ring 2 each independently represent a 6-7 membered aryl group.

65. The method according to claim 51,

wherein, in the formula III:

ring 1 and ring 2 are each independently phenyl.

66. The method according to claim 51,

wherein, in the formula III:

e is trifluoromethyl.

67. The method according to claim 51,

wherein, in the formula III:

m and n are both 2.

68. The use according to claim 51, wherein

In the formula II:

R ^a 、R ^b and R is ^c Each independently selected from hydrogen or methyl.

69. The use according to claim 51, wherein

In the formula II:

is-NH-NH-C (O) -NH-, -N (CH) ₃ ) -NH-C (O) -NH-, -NH-C (O) -NH-or-CH ₂ -NH-C(O)-NH-。

70. The use according to claim 51, wherein

In the formula II:

Ar ¹ is pyridinyl optionally substituted with s groups independently selected from: fluorine, chlorine, bromine, trifluoromethyl, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH ₂ CH ₂ OCH ₃ ，-OCH ₂ CH(OH)CH ₂ OH。

71. The use according to claim 51, wherein

In the formula II:

Ar ¹ is pyridinyl optionally substituted with s groups independently selected from: chloro, bromo, trifluoromethyl, ethoxy, n-propoxy, isopropoxy, n-butoxy, -OCH ₂ CH(OH)CH ₂ OH or-OCH ₂ CH ₂ OCH ₃ 。

72. The use according to claim 51, wherein

In the formula II:

in Ar ¹ The ortho position of the heteroatom in (a) is optionally substituted with 1-2 of said substituents.

73. The use according to claim 51, wherein

In the formula II:

Ar ¹ selected from 4-substituted 2, 6-dichloropyridine, 4-substituted 2, 6-dibromopyridine, 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, 4-substituted-2-trifluoromethyl-6-ethoxypyridine or 4-substituted-2-trifluoromethyl-6- (2, 3-dihydroxypropoxy) pyridine.

74. The use according to claim 51, wherein

In the formula II:

Ar ¹ is 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.

75. The use according to claim 51, wherein

In the formula II:

Ar ¹ And Ar is a group ² Are all 4-substituted 2, 6-dichloropyridine.

76. The method according to claim 51,

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

R ₁ is methyl;

R ₂ selected from hydrogen and methyl.

77. The method according to claim 51,

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

R ₃ and R is ₄ Is not hydrogen and is located at h; and said group located on h is selected from: c optionally substituted by halogen _1-6 An alkyl group; optionally by halogen or C _1-6 Alkoxy substituted phenyl;

the group located on i or j is selected from: hydrogen; c optionally substituted by phenyl _1-6 An alkyl group.

78. The method according to claim 51,

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

R ₃ and R is ₄ Is not hydrogen and is located at h; and said group located on h is selected from: methyl, n-propyl, isopropyl-CF ₃ Phenyl, fluoro-substituted phenyl, methoxy-substituted phenyl;

the group located on i or j is selected from: hydrogen, methyl, n-propyl, isopropyl, benzyl.

79. The method according to claim 51,

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

R ^1a selected from methyl groups.

80. The method according to claim 51,

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

R ^1b Selected from hydrogen.

81. The method according to claim 51,

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

R ³ is methyl.

82. The method according to claim 51,

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

R ⁴ is hydrogen or trifluoromethyl.

83. The method according to claim 51,

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

R ⁴ at least one of which is trifluoromethyl.

84. The method according to claim 51,

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

/>

85. 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 1-31, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of claims 44-49, or a combination of same with one or more other therapeutic agents, or an S1PR2 inhibitor of any one of claims 51-84, wherein the method is performed in vitro.

86. A method for combating/reversing cellular resistance comprising administering to said cells an effective amount of a compound of any one of claims 1-31, a pharmaceutically acceptable salt, or tautomer thereof, or a pharmaceutical composition of any one of claims 44-49, or a combination of same with one or more other therapeutic agents, or an S1PR2 inhibitor of any one of claims 51-84, wherein said method is performed in vitro.

87. The method of claim 85 or 86, wherein the cell is a cell line, or a cell from a subject.

88. The method of claim 85 or 86, wherein the cell is selected from a tumor cell.

89. The method of claim 88, wherein the tumor cell is selected from a breast cancer cell, a liver cancer cell, a lung cancer cell, an esophageal cancer cell, a gastric cancer cell, a pancreatic cancer cell, a cholangiocarcinoma cell, a head and neck tumor cell, a colon cancer cell, or a nasopharyngeal cancer cell.

90. A kit, comprising:

the compound of any one of claims 1-31, a pharmaceutically acceptable salt, or tautomer thereof, or the pharmaceutical composition of any one of claims 44-49; and

optionally an antitumor drug.

91. The kit of claim 90, wherein the anti-tumor drug is selected from fluorouracil-like drugs.