WO2025223036A1 - Mrgprx2 antagonist, pharmaceutical composition comprising same, and use thereof - Google Patents

Abstract

The present invention relates to compounds of formula (I), a pharmaceutical composition comprising same, and the use thereof for preventing or treating diseases that can be improved by means of antagonizing MRGPRX2.

Description

MRGPRX2 antagonists, pharmaceutical compositions comprising them, and their uses Invention Field

This invention relates to MRGPRX2 antagonists, pharmaceutical compositions comprising the same, and their use for the prevention or treatment of diseases that can be improved by antagonizing MRGPRX2.

Background of the Invention

The Mas-related G-protein receptor (MRGPR) family consists of more than 50 members, divided into nine subfamilies: MRGPR A, B, C, D, E, F, G, H, and the primate-specific MRGPRX. Each subfamily includes different subtypes, such as MRGPRX1, MRGPRX2, MRGPRX3, and MRGPRX4. The functions of most of them are still unclear. Human tissues express eight of these receptors, including MRGPR D, E, F, G, and MRGPR X1-4. The MRGPRX receptor is found only in some higher species, including humans. Rodents, however, only express MRGPR A, B, C, and H, and do not express the MRGPRX receptor. The MRGPRB receptor in rodents has a function corresponding to that of the MRGPRX receptor in humans.

MRGPRX2 is primarily expressed in mast cells and some nerve cells. Mast cells are innate immune cells, mainly distributed in areas exposed to the external environment, such as the skin, oral/gastrointestinal mucosa, and respiratory tract. Mast cells are activated primarily through IgE-dependent and non-IgE-dependent pathways, undergoing degranulation and releasing various inflammatory mediators, thereby causing inflammatory and allergic reactions. MRGPRX2 receptors on the mast cell membrane surface can be activated by various mediators, including endogenous neuropeptides (Substance P, Cortistatin-14, etc.) and some exogenous substances (peptides, drugs, microbial derivatives, etc.). MRGPRX2 receptor activation, which induces mast cell degranulation and releases various inflammatory mediators, plays an important role in non-IgE-dependent mast cell activation and participates in the occurrence and development of various inflammatory and allergic diseases. These conditions include chronic urticaria, acute drug hypersensitivity reaction (IDHR)/allergic reactions, atopic dermatitis, chronic pruritus, food allergies, irritable bowel syndrome, allergic rhinitis, nasal polyps, and other conditions related to type II inflammatory response and mast cells.

MRGPRX2 antagonists can stabilize mast cells, block mast cell degranulation caused by MRGPRX2 activation, and prevent downstream inflammation and allergic reactions, thereby playing a therapeutic role in the aforementioned mast cell-related diseases. Furthermore, combination with anti-IgE therapy can also produce additive or complementary effects on mast cell stabilization.

Invention Overview

This invention provides MRGPRX2 antagonists that can be used for the prevention or treatment of diseases that can be improved by antagonizing MRGPRX2. Preferably, the compounds of this invention have antagonistic activity against MRGPRX2. Furthermore, the compounds of this invention also possess superior properties such as better physicochemical properties (e.g., solubility, physical and/or chemical stability), improved pharmacokinetic properties (e.g., improved exposure, bioavailability, suitable half-life and duration of action), and improved safety (lower toxicity and/or fewer side effects).

In one aspect, the present invention provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein said compound has the structure of formula (I):

in:

Ring A, ring B, and ring D are each independently a _C3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring;

The ring C is a _C3-6 hydrocarbon ring or a 3-10 membered heterocycle;

^R1 , ^R2 , ^R5 , and ^R6 are each independently selected from halogens, -OH, _-NH2 , -CN, _-NO2 , _-SF5 , = _CH2 , -CH ^{= CRaRb} , _C1-6 alkyl, deuterated _C1-6 alkyl, halo- _C1-6 alkyl, _C2-6 alkenyl, C2-6 ynyl, _C3-6 cyclic hydrocarbon, _3-10 membered heterocyclic, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Ra , -OC(=O ^{) Ra} , -C(=O) ^ORa _, ^-ORa , -SRa, -S(=O) ^Ra , -S(= ^O ) _2Ra , ^-S (=O) ^2NRaRb , -S(=O)(= ^NRa ) ^Rb -NR ^a R ^b , -C(＝O)NR ^a R ^b , -C(＝O)N(R ^a )-OR ^b , -NR ^a -C(＝O)R ^b , -NR ^a -C(＝O)OR ^b , -NR ^a -S(＝O) ₂ -R ^b , -NR ^a -C(＝O)-NR ^a R ^b , -N＝S(＝O)R ^a R ^b , -P(＝O)R ^a R ^b , -C _1-6 alkylene-R ^a , -C _1-6 alkylene-OR ^a , -C _1-6 alkylene-NR ^a R ^b , -OC _1-6 alkylene-NR ^a R ^b , (-C _3-6 cycloene)-CN and (-C _3-6 cycloene)-C _1-6 alkyl;

Alternatively, when p≥2, two ^R5s on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring.

Alternatively, when q≥2, two ^R6s on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring.

^R3 and ^R4 are each independently selected ^from H, D, -OH, _-NH2 , -CN, -CH= ^CRaRb , _C1-6 alkyl, deuterated _C1-6 alkyl, halogenated _C1-6 alkyl, C2-6 alkenyl, C2-6 ynyl, _C3-6 cycloalkyl, _3-10 heterocyclic, _C6-10 aryl, _5-14 heteroaryl, _C6-12 aralkyl, -C(=O ^{) Ra} , -C(=O) ^ORa , -S ⁽ =O) ^Ra , -S(=O) _2Ra , -S(=O) ^2NRaRb , -S(=O)(= ^{NRa )Rb ,} -C(=O) ^NRaRb , -C(= _O )N( ^Ra ) ^-ORb , _-C1-6 alkylene- ^Ra , -C _1-6 alkylene-OR ^a , -C _1-6 alkylene-NR ^a R ^b , -OC _1-6 alkylene-NR ^a R ^b , (-C _3-6 cycloene)-CN and (-C _3-6 cycloene)-C _1-6 alkyl;

^Ra and ^Rb are each independently selected from H, _C1-6 alkyl, C3-10 cyclic hydrocarbon, _3-10 membered heterocyclic group, _C6-10 aryl, 5-14 membered heteroaryl, and _C6-12 aryl group each time they appear; or ^Ra and ^Rb together with the groups they are attached to form a _C3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a _C6-10 aromatic ring, or a 5-14 membered heteroaryl ring.

The aforementioned alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkylene, cycloalkylene, heterocyclic, heterocyclic, aryl, aromatic, heteroaryl, heteroaromatic, and aralkyl groups are each optionally substituted by one or more substituents independently selected from the following: deuterium, halogen, -OH, =O, _-NH₂ , -CN, _-NO₂ , = _CH₂ , = _CF₂ , -CH= ^CRcRd , _C1-6 alkyl, deuterated _C1-6 alkyl, _C2-6 alkenyl, _C2-6 ^alkynyl , C3-6 cycloalkyl, _3-10 membered heterocyclic, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Rc , -OC(=O) ^Rc , -C(=O) ^ORc , ^-ORc , ^-SRc , -S(=O) ^Rc -S(=O) _₂R₁c , -S(=O ^{) ₂NR₁cR₂d} , -NR₁cR₂d ^{, -C} (=O ^{) NR₁cR₂d} , -NR₁c ^{- C} (=O) ^R₂d , -NR₁c ^- _C (=O) ^OR₂d , -NR₁c ^- S(=O) _₂- ^R₂d , -NR₁c _- ^C (=O ^{) -NR₁cR₂d} , -N=S(=O) ^R₁cR₂d , -C₁ _- 6alkylene- ^OR₁c , -C₁- ^6alkylene - ^NR₁cR₂d and -OC₁- _6alkylene ^{- NR₁cR₂d} , wherein the alkylene, alkyl, alkenyl, = _CH₂ , alkynyl, ^cycloalkyl , heterocyclic, aryl, heteroaryl and aralkyl groups are each optionally further substituted by one or more substituents independently selected from: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH ₂ , -CN, _-NO2 , = _CH2 , = _CF2 , _C1-6 alkyl, _C1-6 haloalkyl, _C3-6 cyclic hydrocarbon, 3-10 heterocyclic, _C6-10 aryl, 5-14 heteroaryl, _C6-12 aralkyl, _-C1-6 alkylene- _C3-6 cyclic hydrocarbon, _-OC1-6 alkyl and _-C1-6 alkylene- _OC1-6 alkyl;

^Rc and ^Rd, each in each occurrence, are independently selected from H, _C1-6 alkyl, C3-10 cyclic hydrocarbon, _3-10 membered heterocyclic group, _C6-10 aryl, 5-14 membered heteroaryl, and _C6-12 aralkyl, or ^Rc and ^Rd together with the groups they are attached to constitute a C3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a _C6-10 aromatic ring, or a 5-14 membered heteroaryl ring. The alkyl, _cyclic hydrocarbon, hydrocarbon ring, heterocyclic group, heterocyclic ring, aryl, aromatic ring, heteroaryl, heteroaryl, and aralkyl groups are further optionally substituted by one or more substituents independently selected from: halogen, -OH, =O, -C(=O)O-tert-butyl, _-NH₂ , -CN, _-NO₂ , _C1-6 alkyl, _C1-6 haloalkyl, C3-6 _cyclic hydrocarbon, 3-10 membered heterocyclic group, C _6-10 aryl, 5-14 heteroaryl, _C6-12 aralkyl and _-C1-6 alkylene- _OC1-6 alkyl;

m, n, p, and q are each independent integers selected from 0, 1, 2, 3, and 4;

The condition is 1) when for and for When p is not 0 or m ≥ 2;

2) When for and for If p is not 0 or at least one ^R2 is _-SF5 .

Another aspect of the invention provides a pharmaceutical composition comprising a preventive or therapeutically effective amount of the compound of the invention or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug, and one or more pharmaceutically acceptable carriers, said pharmaceutical composition preferably being a solid dosage form, a liquid dosage form, or a transdermal dosage form.

Another aspect of the invention provides the use of the compounds of the invention or pharmaceutically acceptable salts, esters, N-oxides, stereoisomers, tautomers, polymorphs, solvates, metabolites, isotopically labeled compounds or prodrugs, or pharmaceutical compositions of the invention, in the preparation of a medicament used as an MRGPRX2 antagonist.

Another aspect of the invention provides compounds of the invention or pharmaceutically acceptable salts, esters, N-oxides, stereoisomers, tautomers, polymorphs, solvates, metabolites, isotopically labeled compounds or prodrugs of the invention, or pharmaceutical compositions of the invention, which are used as MRGPRX2 antagonists.

Another aspect of the invention provides a method for preventing or treating diseases that can be improved by antagonizing MRGPRX2, the method comprising administering to an individual in need an effective amount of the compound of the invention or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug or pharmaceutical composition of the invention.

Invention Details

definition

Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technical terms herein refer to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the invention.

The terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

As used herein, the term "alkylene" means a saturated divalent hydrocarbon group, preferably a saturated divalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, such as methylene, ethylene, propylene or butylene.

As used herein, the term "alkyl" is defined as a straight-chain or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl group has 1 to 12, for example, 1 to 6 carbon atoms. For example, as used herein, the term " _C1-6 alkyl" refers to a linear or branched group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl) _of 1 to 6 carbon atoms, optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogens (in which case the group is referred to as "haloalkyl") (e.g., _CF3 , _C2F5 , _{CHF2 , CH2F , CH2CF3} , _CH2Cl , or _-CH2CH2CF3 , _etc. ). The term “ _C1-4 alkyl” refers to a linear or branched aliphatic hydrocarbon chain with 1 to 4 carbon atoms (i.e., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl).

As used herein, the term "alkenyl" refers to a linear or branched monovalent hydrocarbon group containing one or more double bonds and having 2-6 carbon atoms (" _C2-6 alkenyl"). The alkenyl group is, for example, -CH= _CH2 , _-CH2CH = _CH2 , -C( _CH3 )=CH2, _-CH2 -CH=CH- _CH3 , 2-pentenyl, ₃ -pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, and 4-methyl-3-pentenyl. When the compounds of the present invention contain an alkenyl group, the compounds may exist in pure E (iso-side) form, pure Z (iso-side) form, or any mixture thereof. The term "alkenyl" refers to the corresponding divalent group, including, for example, " _C2-6 alkenyl", " _C2-4 alkenyl", etc. Specific examples include, but are not limited to: -CH=CH-, _-CH2CH =CH-, -C( _CH3 )=CH-, buteneyl, pentenyl, hexeneyl, etc.

As used herein, the term "alkynyl" refers to a monovalent hydrocarbon group comprising one or more triple bonds, preferably having 2, 3, 4, 5, or 6 carbon atoms, such as ethynyl, 2-propynyl, 2-butynyl, 1,3-butadiynyl, etc. The alkynyl group is optionally substituted by one or more (such as 1 to 3) identical or different substituents. The term "ynynyl" refers to the corresponding divalent group, including, for example, "C _2-8 ynynyl,""C _2-6 ynynyl,""C _2-4 ynynyl," etc. Examples include, but are not limited to, [examples not included in the text]. The alkyne group is optionally substituted by one or more (such as 1 to 3) identical or different substituents.

As used herein, the term “fused ring” or “dense ring” refers to a ring system formed by two or more ring structures sharing two adjacent atoms.

As used herein, the term "spiroring" refers to a ring system consisting of two or more ring structures that share a single ring atom.

As used in this article, the term "bridged ring" refers to a ring system formed by two or more ring structures sharing two atoms that are not directly connected to each other.

As used herein, the terms “cycloalkylene group,” “cycloalkylene group,” and “hydrocarbon ring” refer to a saturated (i.e., “cycloalkylene group” and “cycloalkylene group”) or partially unsaturated (i.e., having one or more double and/or triple bonds within the ring) monocyclic or polycyclic hydrocarbon ring (including spirocyclic, fused (fused) ring, or bridged ring systems) having, for example, 3 to 10 (suitably 3 to 8, more preferably 3 to 6) cyclic carbon atoms, including but not limited to (cycloalkylene group) propyl(ring), (cycloalkylene group) butyl(ring), (cycloalkylene group) pentyl(ring), (cycloalkylene group) hexyl(ring), (cycloalkylene group) heptyl(ring), (cycloalkylene group) octyl(ring), (cycloalkylene group) nonyl(ring), (cycloalkylene group) hexenyl(ring), etc.

As used herein, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or bicyclic, including spirocyclic, fused, or bridged systems (e.g., bicyclic [1.1.1]pentyl, bicyclic [2.2.1]heptyl, bicyclic [3.2.1]octyl, or bicyclic [5.2.0]nonyl, decahydronaphthyl, etc.), optionally substituted with one or more (e.g., one to three) suitable substituents. The cycloalkyl group has 3 to 15 carbon atoms. For example, the term "C _3-6 cycloalkyl" refers to a saturated monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) with 3 to 6 cyclic carbon atoms, optionally substituted with one or more (e.g., one to three) suitable substituents, such as methyl-substituted cyclopropyl.

As used herein, the term "heterocyclic group" (or "heterocycle") refers to a saturated or partially unsaturated monocyclic or bicyclic group having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms and one or more (e.g., one, two, three, or four) heteroatoms selected from O, S, N, and P, and the "heterocyclic group" (or "heterocycle") may contain -C (=O)- as a ring member. The heterocyclic group may be attached to the remainder of the molecule via the carbon atoms and/or heteroatoms (if present). Specifically, 3-10 membered heterocyclic groups are groups having 3-10 carbon atoms and heteroatoms in the ring, such as, but not limited to, ethylene oxide, aziridinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, dioxolinyl, pyrrolyl, pyrrolidone, imidazoalkyl, pyrazolyl, pyrrolinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl.

As used herein, the term "heterocyclic group" (or "heterocycle") encompasses fused ring structures, where the connection point between the fused ring structure and other groups can be on any ring within the fused ring structure. Therefore, the heterocyclic groups of the present invention also include, but are not limited to, heterocyclic fused heterocyclic groups, heterocyclic fused cycloalkyl groups, monoheterocyclic fused monoheterocyclic groups, monoheterocyclic fused monocycloalkyl groups, aryl fused heterocyclic groups, and heteroaryl fused heterocyclic groups, such as 3-7 membered (mono)heterocyclic fused 3-7 membered (mono)heterocyclic groups, 3-7 membered (mono)heterocyclic fused (mono)cycloalkyl groups, 3-7 membered (mono)heterocyclic fused _C4-6 (mono)cycloalkyl groups, _C6-10 aryl fused 3-7 membered heterocyclic groups, and 5-6 membered heteroaryl fused 3-7 membered heterocyclic groups. Examples include, but are not limited to, pyrrolidinyl fused cyclopropyl, cyclopentyl fused azacyclopropyl, pyrrolidinyl fused cyclobutyl, pyrrolidinyl fused pyrrolidinyl, pyrrolidinyl fused piperidinyl, pyrrolidinyl fused piperazine, and piperidinyl fused morpholinyl.

As used herein, the term "heterocyclic group" (or "heterocycle") encompasses both bridged heterocyclic groups (bridged heterocycles) and spirocyclic groups (spirocyclic heterocycles).

As used herein, the term "bridged heterocycle" refers to a ring structure containing one or more (e.g., 1, 2, 3, or 4) heteroatoms (e.g., oxygen, nitrogen, and/or sulfur atoms) formed by two rings sharing two non-directly connected ring atoms. This includes, but is not limited to, 7-10 membered bridged heterocycles, 8-10 membered bridged heterocycles, 7-10 membered nitrogen-containing bridged heterocycles, 7-10 membered oxygen-containing bridged heterocycles, 7-10 membered sulfur-containing bridged heterocycles, etc., for example... The "nitrogen-bridged heterocycle", "oxygen-bridged heterocycle", and "sulfur-bridged heterocycle" may optionally also contain one or more other heteroatoms selected from oxygen, nitrogen, and sulfur.

As used herein, the term "spiroheterocycle" refers to a ring structure consisting of two or more rings sharing a single ring atom and containing one or more heteroatoms (e.g., oxygen, nitrogen, sulfur), including but not limited to 5-10 membered spiroheterocycles, 6-10 membered spiroheterocycles, 6-10 membered nitrogen-containing spiroheterocycles, 6-10 membered oxygen-containing spiroheterocycles, 6-10 membered sulfur-containing spiroheterocycles, etc. The "nitrogen-containing spiroheterocycle", "oxygen-containing spiroheterocycle", and "sulfur-containing spiroheterocycle" may optionally also contain one or more other heteroatoms selected from oxygen, nitrogen, and sulfur. The term "6-10-membered nitrogen-containing spiroheterocycle group" refers to a spiroheterocycle group containing a total of 6-10 ring atoms, of which at least one ring atom is a nitrogen atom.

As used herein, the terms “(aryl)aryl” and “aromatic ring” refer to an all-carbon monocyclic or fused-ring polycyclic aromatic group having a conjugated π-electron system. For example, as used herein, the terms “C _6-10 (aryl)aryl” and “C _6-10 aromatic ring” mean an aromatic group containing 6 to 10 carbon atoms, such as (aryl)phenyl (benzene ring) or (aryl)naphthyl (naphthyl ring). The (aryl)aryl and aromatic ring are optionally substituted with one or more (such as 1 to 3) suitable substituents (e.g., halogen, -OH, -CN, _-NO2 , C _1-6 alkyl, etc.).

The term "aralkyl" means an aryl-substituted alkyl group, wherein the aryl group and the alkyl group are as defined herein. Typically, the aryl group may have 6-14 carbon atoms, and the alkyl group may have 1-6 carbon atoms. Exemplary aralkyl groups include, but are not limited to, benzyl, phenylethyl, phenylpropyl, and phenylbutyl.

As used herein, the terms “(sub)heteroaryl” and “heteroary ring” refer to monocyclic, bicyclic, or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 1, 2, 3, 4, 5, 6, 9, or 10 carbon atoms, and containing at least one heteroatom that may be the same or different (the heteroatom being, for example, oxygen, nitrogen, or sulfur), and additionally, in each case, may be benzofused. Specifically, "(hybrid)aryl" or "heteroary ring" is selected from (thienyl)thiophene (ring), (furanyl) (ring), (pyrrolyl) (ring), (oxazolyl) (ring), (thiazolyl) (ring), (imidazolyl) (ring), (pyrazolyl) (ring), (isooxazolyl) (ring), (isothiazolyl) (ring), (oxadiazolyl) (ring), (triazolyl) (ring), (thiadiazolyl) (ring), and their benzo[a] derivatives; or (pyridyl)pyridinyl (ring), (pyridazinyl)pyrimidinyl (ring), (pyrazinyl)pyrazinyl (ring), (triazinyl)pyrazinyl (ring), and their benzo[a] derivatives.

As used herein, the term “halogenated” or “halogenated” is defined as including F, Cl, Br, or I.

As used herein, the term "alkyl thio" means an alkyl group as defined above that is attached to a parent molecule via a sulfur atom. Representative examples of _C1-6 alkyl thio groups include, but are not limited to, methyl thio, ethyl thio, tert-butyl thio, and hexyl thio.

As used herein, the term "nitrogen-containing heterocycle" refers to a saturated or partially unsaturated monocyclic or bicyclic group having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 carbon atoms and at least one nitrogen atom in the ring, and optionally may also contain one or more (e.g., one, two, three, or four) ring members selected from N, O, S, S=O, and S(=O) _₂ ; the nitrogen-containing heterocycle is connected to the remainder of the molecule through any ring member. The nitrogen-containing heterocycle is preferably a saturated nitrogen-containing monocyclic ring. Specifically, 3- to 14-membered nitrogen-containing heterocycles are groups having 3 to 14 carbon atoms and heteroatoms (at least one of which is a nitrogen atom) in the ring, including but not limited to ternary nitrogen-containing heterocycles (such as aziridinyl), quaternary nitrogen-containing heterocycles (such as aziridine), pentazolidinyl, pyrrolinyl, pyrrolidone, imidazolyl, imidazolyl, imidazolinyl, pyrazolyl, pyrazolin ...

The term "substitution" refers to the selective replacement of one or more (e.g., one, two, three, or four) hydrogen atoms on a specified atom by a designated group, provided that the substitution does not exceed the normal valence of the specified atom in the present case and that the substitution forms a stable compound. Combinations of substituents and/or variables are permitted only if such combinations form a stable compound.

If a substituent is described as “optionally substituted,” then the substituent may be (1) unsubstituted or (2) substituted. If the carbon of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the carbon (to the extent that any hydrogens are present) may be substituted individually and/or together with independently selected optional substituents. If the nitrogen of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the nitrogen (to the extent that any hydrogens are present) may each be substituted with independently selected optional substituents.

If a substituent is described as being “independently selected” from a group, then each substituent is selected independently of the others. Therefore, each substituent may be the same as or different from another (other) substituent.

As used herein, the term “one or more” means one or more under reasonable conditions, such as two, three, four, five or ten.

Unless otherwise specified, as used herein, the connection point of a substituent may be located at any suitable position of the substituent.

When the bond of a substituent is such that it passes through the ring and connects two atoms, then such a substituent can be bonded to any cyclic atom in the substituted ring.

This invention also includes all pharmaceutically acceptable isotopically labeled compounds that are identical to the compounds of this invention, except that one or more atoms are replaced by atoms having the same atomic number but a different atomic mass or mass number than the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the compounds of this invention include (but are not limited to) isotopes of hydrogen (e.g., deuterium (D, ^2H ), tritium (T, ^3H )); isotopes of nitrogen (e.g., ^13N and ^15N ); isotopes of oxygen (e.g., ^15O , ^17O , and ^18O ); isotopes of phosphorus (e.g., ^32P ); and isotopes of sulfur (e.g., ^35S ). The isotopically labeled compounds of this invention can be prepared by methods similar to those described in the accompanying routes and/or examples and preparations, using a suitable isotopically labeled reagent instead of the previously used unlabeled reagent. Pharmaceutically acceptable solvates of the present invention include those in which the crystallization solvent may be substituted with an isotope, such as _D₂O , acetone- _d₆ , or DMSO- _d₆ .

The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereomers can be produced. Specific individual molecules can also exist as geometric isomers (cis/trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It is to be understood that the scope of this application covers all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

Solid lines may be used in this article. solid wedge Or virtual wedge The carbon-carbon bonds of the compounds of the present invention are depicted. Solid lines are used to depict bonds to asymmetric carbon atoms to indicate all possible stereoisomers (e.g., specific enantiomers, racemic mixtures, etc.) at that carbon atom. Solid or imaginary wedges are used to depict bonds to asymmetric carbon atoms to indicate the presence of the indicated stereoisomers. When present in racemic mixtures, solid and imaginary wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise specified, the compounds of the present invention are intended to exist as stereoisomers (including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, trans-blocking isomers, and mixtures thereof). The compounds of the present invention may exhibit more than one type of isomerism and may consist of mixtures thereof (e.g., racemic mixtures and diastereomer pairs).

Rotation-restricted isomers are compounds that can be isolated into rotation-restricted isomers.

It should also be understood that certain compounds of the present invention may exist in their free form for therapeutic purposes, or, where appropriate, in their pharmaceutically acceptable derivative forms. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, N-oxides, solvates, metabolites, or prodrugs, which, upon administration to a patient in need, can directly or indirectly provide the compounds of the present invention or their metabolites or residues. Therefore, when referring to "compounds of the present invention" herein, it is also intended to encompass the various derivative forms of the compounds described above.

Pharmaceutically acceptable salts of the compounds of the present invention include their acid addition salts and base addition salts.

For a review of suitable salts, see Stahl and Wermuth's "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

As used herein, the term "ester" means an ester derived from the various general formula compounds of this application, including physiologically hydrolyzable esters (the compounds of the present invention that can be hydrolyzed under physiological conditions to release free acids or alcohols). The compounds of the present invention may themselves also be esters.

Those skilled in the art will understand that not all nitrogen-containing heterocycles can form N-oxides because nitrogen requires available lone pairs of electrons to be oxidized to oxides; those skilled in the art will identify nitrogen-containing heterocycles that can form N-oxides. Those skilled in the art will also recognize that tertiary amines can form N-oxides. Synthetic methods for preparing N-oxides of heterocycles and tertiary amines are well known to those skilled in the art, including the oxidation of heterocycles and tertiary amines with peroxyacids such as peracetic acid and m-chloroperoxybenzoic acid (m-CPBA), hydrogen peroxide, alkyl peroxides such as tert-butyl peroxide, sodium perborate, and dioxiranes such as dimethyldioxirane. These methods for preparing N-oxides have been extensively described and reviewed in the literature, see, for example: T.L. Gilchrist, Comprehensive Organic Synthesis, vol.7, pp 748-750; S.V. Ley, Ed., Pergamon Press; M. Tisler and B. Stanovnik, Comprehensive Heterocyclic Chemistry, vol.3, pp 18-20.

The compounds of the present invention can exist as solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, can be stoichiometric or non-stoichiometric.

The scope of this invention also includes metabolites of the compounds of this invention, i.e., substances formed in the body when the compounds of this invention are administered. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of this invention, including compounds obtained by methods that expose the compounds of this invention to mammals for a time sufficient to produce their metabolites.

This invention further includes, within its scope, prodrugs of the compounds of the invention, which are certain derivatives of the compounds of the invention that may themselves have little or no pharmacological activity, which, when administered to or onto the body, can be converted, for example, by hydrolysis and cleavage into the compounds of the invention having the desired activity. Typically, such prodrugs are functional group derivatives of the compounds that are readily converted in vivo into the desired therapeutically active compounds. Further information regarding the use of prodrugs can be found in “Pro-drugs as Novel Delivery Systems,” Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella) and “Bioreversible Carriers in Drug Design,” Pergamon Press, 1987 (edited by E.B. Roche, American Pharmaceutical Association). The prodrug of the present invention can be prepared, for example, by replacing suitable functional groups present in the compounds of the present invention with certain portions known to those skilled in the art as “pro-moiety” (e.g., “Design of Prodrugs”, as described in H. Bundgaard (Elsevier, 1985)).

This invention also covers compounds of the invention containing protecting groups. In any process of preparing the compounds of the invention, protection of sensitive or reactive groups on any relevant molecule may be necessary and/or desired, thereby forming a form of chemical protection for the compounds of the invention. This can be achieved by conventional protecting groups, for example, those described in *Protective Groups in Organic Chemistry*, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P.G.M. Wuts, *Protective Groups in Organic Synthesis*, John Wiley & Sons, 1991, which are incorporated herein by reference. Protecting groups can be removed at appropriate subsequent stages using methods known in the art.

As used herein, the term “about” means within ±10% of the stated value, preferably within ±5%, and more preferably within ±2%.

compound

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein said compound has the structure of formula (I):

in:

Ring A, ring B, and ring D are each independently a _C3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring;

The ring C is a _C3-6 hydrocarbon ring or a 3-10 membered heterocycle;

^R1 , ^R2 , ^R5 , and ^R6 are each independently selected from halogens, -OH, _-NH2 , -CN, _-NO2 , _-SF5 , = _CH2 , -CH ^{= CRaRb} , _C1-6 alkyl, deuterated _C1-6 alkyl, halo- _C1-6 alkyl, _C2-6 alkenyl, C2-6 ynyl, _C3-6 cyclic hydrocarbon, _3-10 membered heterocyclic, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Ra , -OC(=O ^{) Ra} , -C(=O) ^ORa _, ^-ORa , -SRa, -S(=O) ^Ra , -S(= ^O ) _2Ra , ^-S (=O) ^2NRaRb , -S(=O)(= ^NRa ) ^Rb -NR ^a R ^b , -C(＝O)NR ^a R ^b , -C(＝O)N(R ^a )-OR ^b , -NR ^a -C(＝O)R ^b , -NR ^a -C(＝O)OR ^b , -NR ^a -S(＝O) ₂ -R ^b , -NR ^a -C(＝O)-NR ^a R ^b , -N＝S(＝O)R ^a R ^b , -P(＝O)R ^a R ^b , -C _1-6 alkylene-R ^a , -C _1-6 alkylene-OR ^a , -C _1-6 alkylene-NR ^a R ^b , -OC _1-6 alkylene-NR ^a R ^b , (-C _3-6 cycloene)-CN and (-C _3-6 cycloene)-C _1-6 alkyl;

Alternatively, when p≥2, two ^R5s on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring.

Alternatively, when q≥2, two ^R6s on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring.

^R3 and ^R4 are each independently selected ^from H, D, -OH, _-NH2 , -CN, -CH= ^CRaRb , _C1-6 alkyl, deuterated _C1-6 alkyl, halogenated _C1-6 alkyl, C2-6 alkenyl, C2-6 ynyl, _C3-6 cycloalkyl, _3-10 heterocyclic, _C6-10 aryl, _5-14 heteroaryl, _C6-12 aralkyl, -C(=O ^{) Ra} , -C(=O) ^ORa , -S ⁽ =O) ^Ra , -S(=O) _2Ra , -S(=O) ^2NRaRb , -S(=O)(= ^{NRa )Rb ,} -C(=O) ^NRaRb , -C(= _O )N( ^Ra ) ^-ORb , _-C1-6 alkylene- ^Ra , -C _1-6 alkylene-OR ^a , -C _1-6 alkylene-NR ^a R ^b , -OC _1-6 alkylene-NR ^a R ^b , (-C _3-6 cycloene)-CN and (-C _3-6 cycloene)-C _1-6 alkyl;

^Ra and ^Rb are each independently selected from H, _C1-6 alkyl, C3-10 cyclic hydrocarbon, _3-10 membered heterocyclic group, _C6-10 aryl, 5-14 membered heteroaryl, and _C6-12 aryl group each time they appear; or ^Ra and ^Rb together with the groups they are attached to form a _C3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a _C6-10 aromatic ring, or a 5-14 membered heteroaryl ring.

The aforementioned alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkylene, cycloalkylene, heterocyclic, heterocyclic, aryl, aromatic, heteroaryl, heteroaromatic, and aralkyl groups are each optionally substituted by one or more substituents independently selected from the following: deuterium, halogen, -OH, =O, _-NH₂ , -CN, _-NO₂ , = _CH₂ , = _CF₂ , -CH= ^CRcRd , _C1-6 alkyl, deuterated _C1-6 alkyl, _C2-6 alkenyl, _C2-6 ^alkynyl , C3-6 cycloalkyl, _3-10 membered heterocyclic, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Rc , -OC(=O) ^Rc , -C(=O) ^ORc , ^-ORc , ^-SRc , -S(=O) ^Rc -S(=O) _₂R₁c , -S(=O ^{) ₂NR₁cR₂d} , -NR₁cR₂d ^{, -C} (=O ^{) NR₁cR₂d} , -NR₁c ^{- C} (=O) ^R₂d , -NR₁c ^- _C (=O) ^OR₂d , -NR₁c ^- S(=O) _₂- ^R₂d , -NR₁c _- ^C (=O ^{) -NR₁cR₂d} , -N=S(=O) ^R₁cR₂d , -C₁ _- 6alkylene- ^OR₁c , -C₁- ^6alkylene - ^NR₁cR₂d and -OC₁- _6alkylene ^{- NR₁cR₂d} , wherein the alkylene, alkyl, alkenyl, = _CH₂ , alkynyl, ^cycloalkyl , heterocyclic, aryl, heteroaryl and aralkyl groups are each optionally further substituted by one or more substituents independently selected from: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH ₂ , -CN, _-NO2 , = _CH2 , = _CF2 , _C1-6 alkyl, _C1-6 haloalkyl, _C3-6 cyclic hydrocarbon, 3-10 heterocyclic, _C6-10 aryl, 5-14 heteroaryl, _C6-12 aralkyl, _-C1-6 alkylene- _C3-6 cyclic hydrocarbon, _-OC1-6 alkyl and _-C1-6 alkylene- _OC1-6 alkyl;

^Rc and ^Rd, each in each occurrence, are independently selected from H, _C1-6 alkyl, C3-10 cyclic hydrocarbon, _3-10 membered heterocyclic group, _C6-10 aryl, 5-14 membered heteroaryl, and _C6-12 aralkyl, or ^Rc and ^Rd together with the groups they are attached to constitute a C3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a _C6-10 aromatic ring, or a 5-14 membered heteroaryl ring. The alkyl, _cyclic hydrocarbon, hydrocarbon ring, heterocyclic group, heterocyclic ring, aryl, aromatic ring, heteroaryl, heteroaryl, and aralkyl groups are further optionally substituted by one or more substituents independently selected from: halogen, -OH, =O, -C(=O)O-tert-butyl, _-NH₂ , -CN, _-NO₂ , _C1-6 alkyl, _C1-6 haloalkyl, C3-6 _cyclic hydrocarbon, 3-10 membered heterocyclic group, C _6-10 aryl, 5-14 heteroaryl, _C6-12 aralkyl and _-C1-6 alkylene- _OC1-6 alkyl;

m, n, p, and q are each independent integers selected from 0, 1, 2, 3, and 4;

The condition is 1) when for and for When p is not 0 or m ≥ 2;

2) When for and for If p is not 0 or at least one ^R2 is _-SF5 .

In a preferred embodiment, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound has a structure of formula (II), (III) or (IV):

in

p’ is 0, 1 or 2;

The remaining groups are as defined in this document.

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ring A and ring B are each independently a _C6-10 aromatic ring or a 5-14 membered heteroaromatic ring.

In a preferred embodiment, ring A and ring B are each independently a benzene ring or a 5-6 membered heteroaromatic ring.

In a preferred embodiment, Selected from

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R1 and ^R2, each in each occurrence, are independently selected from halogens, -CN, _-SF5 , _C1-6 alkyl, halogenated _C1-6 alkyl, and -O-( _C1-6 alkyl).

In a preferred embodiment, ^R1 and ^R2 are each independently selected from F, Cl, CN, _-SF5 , _CHF2 , _CF3 and _-OCH3 each time they appear.

In a preferred embodiment, ^R1 and ^R2 are each independently selected from F, Cl, CN, _-SF5 , _CF3 and _-OCH3 each time they appear.

In a preferred embodiment, ^R1 is a halogenated _C1-6 alkyl group, preferably _CF3 .

In a preferred embodiment, ^R2 is a halogen, preferably Cl.

In the preferred embodiment, m is 1.

In the preferred embodiment, n is 1.

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R3 and ^R4 are both H.

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein when for and for When p≥2, the two ^R5s located on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a benzene ring, or a 5-14 membered heteroaromatic ring; preferably, at this time, for

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R5 is independently _C1-6 alkyl each time it appears.

In some implementations, when p≥2, two ^R5s on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-6 membered heterocycle, a benzene ring, or a 5-6 membered heteroaromatic ring.

In some embodiments, when p≥2, two R5 ^atoms located on the same or different ring atoms together constitute a _C1-4 alkylene, preferably a _C1-3 alkylene, and more preferably an ethylene.

In some embodiments, p=2, the two ^R5s are located on different ring atoms, and the two ^R5s together constitute a _C1-4 alkylene, preferably a _C1-3 alkylene, and more preferably an ethylene.

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein Selected from

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ring D is selected from 5-6 membered heterocycles, benzene rings or 5-6 membered heteroaromatic rings.

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ring D is selected from a benzene ring or a 5-6 membered heteroaromatic ring.

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R6 , each time it appears, is independently a halogen, _-NH2 , -CN, _C1-6 alkyl, halo- _C1-6 alkyl, _C3-6 cycloalkyl, phenyl, -C(=O)-( _C1-6 alkyl), -O- _(C1-6 alkyl), -NH( _C1-6 alkyl), -NH(halo- _C1-6 alkyl), -NH-S(=O) _2- ( _C1-6 alkyl), _-C1-6 alkylene-CN, or _-C1-6 alkylene-OH.

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R6 , each time it appears, is independently a halogen, _-NH2 , -CN, _C1-6 alkyl, halo- _C1-6 alkyl, _C3-6 cycloalkyl, phenyl, -O-( _C1-6 alkyl), -NH( _{C1-6 alkyl), -NH(halo-C1-6 alkyl} ), -NH-S(=O) _2- ( _C1-6 alkyl), _-C1-6 alkylene-CN, or _-C1-6 alkylene-OH.

In some implementations, when q≥2, two ^R6s on the same or different ring atoms, together with the groups they are attached to, constitute a benzene ring or a 5-6 membered heteroaromatic ring.

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein Selected from

In some embodiments, this disclosure provides a compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein Selected from

This disclosure covers the technical solutions/compounds obtained by arbitrarily combining any two or more of the above embodiments.

In a preferred embodiment, this disclosure provides the above-described compounds or their pharmaceutically acceptable salts, esters, N-oxides, stereoisomers, tautomers, polymorphs, solvates, metabolites, isotopically labeled compounds or prodrugs, wherein the compounds are selected from:

Pharmaceutical compositions and treatment methods

In some embodiments, the present invention provides a pharmaceutical composition comprising a preventive or therapeutically effective amount of the compound of the present invention or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug, and one or more pharmaceutically acceptable carriers, said pharmaceutical composition preferably being a solid dosage form, a liquid dosage form, or a transdermal dosage form.

In some embodiments, the present invention provides the use of the compounds of the present invention or pharmaceutically acceptable salts, esters, N-oxides, stereoisomers, tautomers, polymorphs, solvates, metabolites, isotopically labeled compounds or prodrugs of the present invention, or pharmaceutical compositions of the present invention, in the preparation of a medicament used as an MRGPRX2 antagonist.

In some embodiments, the present invention provides compounds of the present invention or pharmaceutically acceptable salts, esters, N-oxides, stereoisomers, tautomers, polymorphs, solvates, metabolites, isotopically labeled compounds or prodrugs of the present invention, or pharmaceutical compositions of the present invention, which are used as MRGPRX2 antagonists.

In some embodiments, the present invention provides a method for preventing or treating diseases that can be improved by antagonizing MRGPRX2, the method comprising administering to an individual in need an effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug or pharmaceutical composition of the present invention.

The conditions that can be improved by antagonizing MRGPRX2 include, but are not limited to, urticaria, drug-induced acute allergic/anaphylactic reactions, asthma, dermatitis (e.g., atopic dermatitis), pruritus, allergies (e.g., food allergies), enteritis, irritable bowel syndrome, arthritis, allergic rhinitis, nasal polyps, and other conditions related to type II inflammatory response and mast cells.

In this invention, "pharmaceutically acceptable carrier" refers to a diluent, excipient, vehicle, or medium that is administered co-administered with a therapeutic agent and is suitable, to the extent of reasonable medical judgment, for contact with human and/or other animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit/risk ratio.

Pharmaceutically acceptable carriers that can be used in the pharmaceutical compositions of the present invention include, but are not limited to, sterile liquids such as water and oils, including those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is an exemplary carrier when the pharmaceutical composition is administered intravenously. Physiological saline and aqueous solutions of glucose and glycerol can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene glycol, water, ethanol, etc. The compositions may also contain small amounts of wetting agents, emulsifiers, or pH buffers as needed. Oral formulations may contain standard carriers such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutically acceptable carriers are described in Remington’s Pharmaceutical Sciences (1990).

The pharmaceutical compositions of the present invention can act systemically and/or locally. For this purpose, they can be administered via suitable routes, such as by injection (e.g., intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular injection, including infusion) or transdermal administration; or by oral, sublingual, nasal, transmucosal, topical, ophthalmic formulations or by inhalation.

For these routes of administration, the pharmaceutical compositions of the present invention can be administered in suitable dosage forms.

The dosage forms include, but are not limited to, tablets, capsules, lozenges, hard candies, powders, sprays, creams, ointments, suppositories, gels, pastes, lotions, ointments, aqueous suspensions, injectable solutions, elixirs, and syrups.

As used in this article, the term "effective amount" refers to the amount of a compound that, when administered, will alleviate one or more symptoms of the treated condition to some extent.

The dosing regimen can be adjusted to provide the optimal required response. For example, a single bolus injection can be administered, several fractions can be administered over time, or the dose can be proportionally reduced or increased as indicated by the urgency of the treatment situation. It should be noted that dosage values can vary depending on the type and severity of the condition to be alleviated, and may include single or multiple doses. To further understand, for any given individual, the specific dosing regimen should be adjusted over time based on individual needs and the professional judgment of the person administering the composition or supervising its administration.

The amount of the compounds of the present invention administered will depend on the individual being treated, the severity of the condition or illness, the rate of administration, the disposal of the compounds, and the prescribing physician's judgment. Generally, the effective dose is from about 0.0001 to about 50 mg per kg of body weight per day, for example, from about 0.01 to about 10 mg/kg/day (single or divided doses). For a 70 kg person, this would total from about 0.007 mg/day to about 3500 mg/day, for example, from about 0.7 mg/day to about 700 mg/day. In some cases, dose levels not exceeding the lower limit of the foregoing range may be sufficient, while in other cases, larger doses may still be used without causing any harmful side effects, provided that the larger dose is first divided into several smaller doses administered throughout the day.

The content or amount of the compound of the present invention in the pharmaceutical composition may be from about 0.01 mg to about 1000 mg, preferably 0.1-500 mg, preferably 0.5-300 mg, more preferably 1-150 mg, particularly preferably 1-50 mg, such as 1.5 mg, 2 mg, 4 mg, 10 mg, 25 mg, etc.

Unless otherwise stated, as used herein, the term “treatment” means to reverse, alleviate, or inhibit the progression of a disease or condition or one or more symptoms of such a disease or condition to which such term is applied, or to prevent such a disease or condition or one or more symptoms of such a disease or condition.

As used herein, “individual” includes both human and non-human animals. Exemplary human individuals include human individuals suffering from a disease (such as the disease described herein) (referred to as patients) or normal individuals. In this invention, “non-human animals” includes all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock, and/or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

In another embodiment, the pharmaceutical composition of the present invention may also contain one or more additional therapeutic or preventative agents.

Example

The present invention is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the invention.

Experimental methods in this invention, where specific conditions are not specified, are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. Reagents not specifically sourced are commercially available conventional reagents. All evaporations were performed using a rotary evaporator under vacuum. Analytical samples were dried under vacuum (1-5 mmHg) at room temperature. Separation was performed using pre-p-TLC or high-performance liquid chromatography (HPLC) on preparative silica gel plates. Rapid column chromatography purification was performed using SEPAFLASH pre-packed silica gel columns, and mixed solvent systems were reported as volume ratios.

The structure of the compound was confirmed by nuclear magnetic resonance spectroscopy (NMR) or mass spectrometry (MS).

NMR spectra were recorded using a Varian NMR System 400 MHz high-resolution NMR spectra. Chemical shifts (δ) are given in parts per million (ppm). The solvents used for determination were deuterated chloroform ( _CDCl₃ ), hexadeuterated dimethyl sulfoxide (DMSO-d₆), or deuterated methanol ( _CD₃OD ). The internal standard was tetramethylsilane (TMS). The abbreviations for the splitting multiples of the ^¹H NMR peaks are as follows: s for singlet, bs for broad singlet, d for doublet, t for triplet, q for quartet, m for multiplet, dd for doublet, etc.

The liquid chromatography-mass spectrometry (LC-MS) instrument used was an Agilent 1260 series 6135 mass spectrometer with electrospray ionization capability. The analytical method is as follows:

Agilent LC-MS 1260-6135, column: Agilent ZORBAX SB-C18 (50 mm × 2.1 mm × 5 μm); column temperature: 25 °C; flow rate: 1.5 mL/min; mobile phase: changed from a mixture of 95% [water + 0.1% trifluoroacetic acid] and 5% [acetonitrile + 0.1% trifluoroacetic acid] to a mixture of 5% [water + 0.05% trifluoroacetic acid] and 95% [acetonitrile + 0.05% trifluoroacetic acid] within 2.5 min.

The high-performance liquid chromatograph used was a Hanbang 50ml binary semi-preparative liquid chromatography system, with a column type of Hedera ODS-2 250*10mm*10μm; the mobile phases were: phase A [water + 0.1% trifluoroacetic acid] and phase B [acetonitrile + 0.1% trifluoroacetic acid].

The thin-layer chromatography silica gel plates used were Huanghai GF254 silica gel plates.

Unless otherwise specified in the examples, all reactions were carried out under a nitrogen atmosphere.

The reaction process in the examples was monitored using thin-layer chromatography (TLC) or liquid chromatography-mass spectrometry (LC-MS).

The abbreviations used in this invention have the following meanings:

Example 1: Preparation of Compound 1

Step 1. Preparation of Compound 1-1

4,6-Dichloro-2-(trifluoromethyl)quinoline (180 mg), (3S,5R)-5-aminotetrahydro-2H-pyran-3-ol hydrochloride (100 mg), and N,N-diisopropylethylamine (0.35 g) were dissolved in anhydrous dimethyl sulfoxide (3 mL) and reacted at 130 °C for 3 hours. The reaction solution was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: petroleum ether-ethyl acetate = 1:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a white solid 1-1 (95 mg, yield 40%). MS-ESI m/z: 347.1 [M+H]+.

Step 2. Preparation of compounds 1-2

Dissolve 1-1 (95 mg) in anhydrous dichloromethane (5 mL), add Dys-Martin oxidant (340 mg), react at room temperature for 2 hours, dilute with saturated sodium bicarbonate aqueous solution, extract with dichloromethane, collect the organic phase, dry with anhydrous sodium sulfate, filter, concentrate, and purify by column chromatography [eluent: petroleum ether-ethyl acetate = 1:1]. Collect the eluent, remove the solvent under reduced pressure, and give a white solid 1-2 (80 mg, yield 85%). MS-ESI m/z: 345.1 [M+H]+.

Step 3. Preparation of compounds 1-3

1-2 (80 mg), tert-butyl carbamate (81 mg), triethylsilane (80 mg), and trifluoroacetic acid (52 mg) were dissolved in anhydrous acetonitrile (3 mL) and reacted overnight at 40 °C. The reaction solution was concentrated to dryness, diluted with dichloromethane, and the organic phase was washed with saturated sodium bicarbonate aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: petroleum ether-ethyl acetate = 1:1]. The eluent was collected, and the solvent was removed by vacuum distillation to give a white solid 1-3 (30 mg, yield 37%). MS-ESI m/z: 346.1 [M+H]+.

Step 4. Preparation of Compound 1

1-3 (20 mg), 1-methylpyrazole-4-carboxylic acid (11 mg), HATU (44 mg), and N,N-diisopropylethylamine (30 mg) were dissolved in anhydrous N,N-dimethylformamide (2 mL) and reacted overnight at room temperature. The reaction solution was diluted with water, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative thin-layer chromatography [eluent: dichloromethane-methanol = 20:1]. The eluent was collected, and the solvent was removed by vacuum distillation to give a white solid 1 (1.1 mg, yield 4%). MS-ESI m/z:454.1[M+H]+. ¹ H NMR (400MHz, CDCl ₃ )δ8.04(d,J＝8.8Hz,1H),7.87(s,1H),7.74(s,1H),7.68(m,2H),7.23(s ,1H),4.20(m,2H),3.91(s,3H),3.86(m,1H),1.63(m,4H),1.53(m,1H).

Example 7: Preparation of Compound 7

Step 1. Preparation of compound 7-1

4,6-Dichloro-2-(trifluoromethyl)quinoline (80 mg) and (5-aminobicyclo[3.2.1]octane-1-yl)carbamate (91 mg) were dissolved in dimethyl sulfoxide (2 ml). The mixture was stirred at 130 °C for 4 hours. After the reaction was completed, the mixture was diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. The organic phase was purified by column chromatography [eluent: petroleum ether-ethyl acetate = 2:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a white solid 7-1 (80 mg, yield 53%). MS-ESI m/z: 504.1 [M+H] ⁺ .

Step 2. Preparation of compound 7-2

Dissolve 7-1 (60 mg) in methanol (5 ml), add zinc chloride (30 mg) and palladium on carbon (10% wt, 20 mg), and stir at room temperature for 6 hours under a hydrogen balloon. Filter and concentrate the reaction solution, then purify by column chromatography [eluent: dichloromethane-methanol = 10:1]. Collect the eluent, remove the solvent under reduced pressure, and give a colorless oily 7-2 (30 mg, yield 68%). MS-ESI m/z: 370.1 [M+H] ⁺ .

Step 3. Preparation of Compound 7

7-2 (30 mg), 1-methylpyrazol-4-carboxylic acid (13 mg), N,N-diisopropylethylamine (31 mg), and HATU (46 mg) were dissolved in N,N-dimethylformamide (5 ml) and stirred at room temperature for 1 hour. The solution was diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. The solution was purified by column chromatography [eluent:eluent:dichloromethane-methanol = 15:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a white solid 7 (5 mg, yield 13%). MS-ESI m/z: 478.1 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ )δ8.65(s,1H),8.08(s,1H),7.84-7.80(m,2H),7.72(d,J＝8.9Hz,1H),7.55(s,1H),7.34(s,1H),6.78 (s,1H),3.81(s,3H),2.26(d,J＝10.2Hz,2H),2.18–1.87(m,4H),1.76(d,J＝31.9Hz,4H),1.50(s,2H).

The following compounds were synthesized using a route similar to that in Example 7:

Example 27: Preparation of Compound 27

Step 1. Preparation of compound 27-1

2,2-Dimethyl-N-(4-pyridyl)propionamide (2 g) was dissolved in tetrahydrofuran (20 mL), and n-butyllithium (1.6 M, 17.5 mL) was added dropwise at -78 °C. The reaction was carried out at -20 °C for 3 hours, followed by the addition of diethyl oxalate (4.0 mL) at -78 °C. After reacting at -78 °C for 15 minutes, the temperature was raised to room temperature. Once the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: petroleum ether-ethyl acetate = 1:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a yellow oily 27-1 (1.05 g, yield 34%). MS-ESI m/z: 279.1 [M+H]+.

Step 2. Preparation of compound 27-2

Dissolve 27-1 (800 mg) in water (4 mL) and ethanol (1 mL), add potassium hydroxide (668 mg), and react at 100 °C for 2 hours. After the reaction is complete, cool to room temperature, add 1,1,1-trifluoroacetone (3.3 g), and react at 100 °C for 5 hours. After the reaction is complete, cool to room temperature, concentrate, extract with dichloromethane, adjust the pH of the aqueous phase to 1 with dilute hydrochloric acid (1 N), extract with ethyl acetate, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate, and give a white solid 27-2 (30 mg, yield 5%). MS-ESI m/z: 243.1 [M+H]+.

Step 3. Preparation of compound 27-3

27-2 (30 mg) was dissolved in toluene (2 mL), and tert-butanol (18 mg), triethylamine (36 mg), and DPPA (64 mg) were added. The reaction was carried out at 110 °C for 1.5 hours. After the reaction was completed, the solution was concentrated, diluted with water, extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. The mixture was stirred at room temperature for 1 hour, and the pH was adjusted to 10–13 with concentrated ammonia. The solution was extracted with dichloromethane, and the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow oily 27-3 (25 mg, 95% yield). MS-ESI m/z: 214.1 [M+H]+.

Step 4. Preparation of compound 27-4

27-3 (25 mg) was dissolved in acetonitrile (5 mL), and cuprous iodide (21 mg) and amyl nitrite (21 mg) were added. The mixture was reacted at 70 °C for 2 hours. The reaction solution was filtered and concentrated, diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by column chromatography [eluent: dichloromethane-methanol = 20:1]. The eluent was collected, concentrated, and a white solid 27-4 (13 mg, yield 48%) was obtained. MS-ESI m/z: 233.1 [M+H]+.

Step 5. Preparation of compound 27-5

27-4 (10 mg) was dissolved in anhydrous dimethyl sulfoxide (2 mL), and tert-butyl ((1R,3S)-3-aminocyclohexyl)carbamate (10 mg) and N,N-diisopropylethylamine (22 mg) were added. The mixture was reacted at 130 °C for 2 hours, cooled to room temperature, diluted with water, extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: dichloromethane-methanol = 20:1]. The eluent was collected, concentrated, and a pale yellow oil 27-5 (15 mg, yield 85%) was obtained. MS-ESI m/z: 411.2 [M+H]+.

Step 6. Preparation of compound 27-6

27-5 (15 mg) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (1 mL) was added. The mixture was stirred at room temperature for 30 minutes. After the reaction was complete, concentrated ammonia was added to adjust the pH to approximately 10. The solution was diluted with water, extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a colorless oily 27-6 (10 mg, yield 88%). MS-ESI m/z: 311.1 [M+H]+.

Step 7. Preparation of Compound 27

27-6 (10 mg) was dissolved in N,N-dimethylformamide (1 mL), and 1-methylpyrazol-4-carboxylic acid (5 mg), HATU (18 mg), and N,N-diisopropylethylamine (16 mg) were added. The reaction mixture was reacted at room temperature for 1 hour. The reaction solution was diluted with water, extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by column chromatography [eluent: dichloromethane-methanol = 15:1]. The eluent was collected, concentrated, and a pale yellow solid 27 (6.0 mg, yield 42%) was obtained. MS-ESI m/z: 419.2 [M+H]+. 1H NMR (400MHz, DMSO-d ₆ )δ9.74(s,1H),8.62(dd,J＝5.9,1.3Hz,1H),8.07(s,1H),7.99(d,J＝8.1Hz,1H),7.87(d,J＝7.9Hz,1H),7.79(s,1H),7.66(d,J＝5.9 Hz,1H),7.01(s,1H),3.98–3.87(m,2H),3.79(s,3H),2.12–2.04(m,1H),1.96–1.88(m,1H),1.84–1.72(m,2H),1.54–1.31(m,4H).

The following compounds were synthesized using a route similar to that in Example 27:

Example 35: Preparation of compound 35

Step 1. Preparation of compound 35-1

5-Bromo-2-(trifluoromethyl)imidazo[1,2-a]pyridine (80 mg), cis-3-amino-1-cyclobutylcarbamate tert-butyl ester (60 mg), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (BINAP) (37 mg), sodium tert-butoxide (86 mg), and tris(dibenzylacetone)palladium (27 mg) were dissolved in 1,4-dioxane (2 ml) and reacted at 85 °C for 2 hours. The reaction solution was diluted with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: petroleum ether-ethyl acetate = 2:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a white solid 35-1 (40 mg, yield 35%). MS-ESI m/z: 371.1 [M+H]+.

Step 2. Preparation of compound 35-2

35-1 (60 mg) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (1 mL) and reacted at room temperature for half an hour. The reaction solution was concentrated, diluted with water, and the pH was adjusted to ~10 with ammonia. The solution was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: dichloromethane-methanol = 10:1]. The eluent was collected, and the solvent was removed by vacuum distillation to give a colorless oily 35-2 (30 mg, yield 70%). MS-ESI m/z: 271.1 [M+H]+.

Step 3. Preparation of compound 35

35-2 (25 mg), p-methoxybenzoic acid (15 mg), N,N-diisopropylethylamine (36 mg), and HATU (42 mg) were dissolved in N,N-dimethylformamide (3 mL) and reacted at room temperature for 1 hour. The reaction solution was diluted with water, extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: dichloromethane-methanol = 10:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a white solid 35 (11 mg, yield 29%). MS-ESI m/z: 405.1 [M+H]+. ¹ H NMR (500MHz, CDCl ₃ )δ8.01(s,1H),7.81(d,J＝8.4Hz,1H),7.72–7.66(m,2H),7.50–7.37(m,2H),6.96(d,J＝1.3Hz,1H),6.95–6.90(m,2 H),6.85(d,J=6.6Hz,1H),4.15–4.04(m,1H),3.88–3.86(m,1H),3.83(s,3H),2.09–2.00(m,2H),1.81–1.72(m,2H).

The following compounds were synthesized using a route similar to that in Example 35:

Example 37: Preparation of compound 37

Step 1. Preparation of compound 37-1

2-Amino-4-bromothiazole (2 g) and 3-bromo-1,1,1-trifluoroacetone (4.27 g) were dissolved in anhydrous 1,4-dioxane (20 mL) and reacted at 110 °C for 12 hours. The mixture was filtered through diatomaceous earth, washed with ethyl acetate, and the organic phase was washed with saturated sodium bicarbonate aqueous solution. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: petroleum ether-ethyl acetate = 1:1]. The eluent was collected, and the solvent was removed by vacuum distillation to give a brown oily substance 37-1 (600 mg, yield 20%). MS-ESI m/z: 270.9 [M+H]+.

Step 2. Preparation of compound 37-2

37-1 (260 mg), 1-N-Boc-cis-1,4-cyclohexanediamine (270 mg), [2-(dicyclohexylphosphine)-3-tert-butoxy-6-methoxy-2',6'-diisopropyl-1,1'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-1-yl)palladium bromide (Gphos Pd G6) (192 mg), and sodium trimethylsilanolate (120 mg) were dissolved in anhydrous tetrahydrofuran (5 mL) and reacted at 50 °C for 3 hours. After filtration through diatomaceous earth and washing with ethyl acetate, the organic phase was washed with saturated brine, the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by column chromatography [eluent: petroleum ether-ethyl acetate = 4:1]. The eluent was collected, and the solvent was removed by vacuum distillation to give a brown oily substance 37-2 (180 mg, yield 46%). MS-ESI m/z:405.2[M+H]+.

Step 3. Preparation of compound 37-3

37-2 (180 mg) was dissolved in dichloromethane (2 mL), and under ice-water bath conditions, trifluoroacetic acid (0.4 mL) was added. The reaction was carried out for 30 minutes, concentrated, and the pH was adjusted to 7-8 with saturated sodium bicarbonate aqueous solution. The mixture was extracted with dichloromethane, and the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude brown oily product 37-3 (0.13 g), which was directly used in the next reaction. MS-ESI m/z: 305.1 [M+H]+.

Step 4. Preparation of compound 37

37-3 (20 mg), p-methoxybenzoic acid (15 mg), HATU (50 mg), and N,N-diisopropylethylamine (34 mg) were dissolved in anhydrous N,N-dimethylformamide (2 mL) and reacted at room temperature. After the reaction was complete, the solution was diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: dichloromethane-methanol = 20:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a white solid 37 (5.0 mg, yield 17%). MS-ESI m/z: 439.1 [M+H]+. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.05–7.98(m,2H),6.95(d,J＝8.6Hz,4H),6.26(d,J＝4.5Hz,1H),5.71(s,1H),3.83(m,1 H),3.78(s,3H),3.43(m,1H),2.01–1.91(m,2H),1.79(d,J＝9.5Hz,2H),1.73–1.67(m,4H).

The following compounds were synthesized using a route similar to that in Example 37:

Examples 100 and 101:

Step 1.

80 (210 mg) was separated by chiral HPLC [CHIRALPAK IB N-5, 5.0 cm ID. x 25 cm L. 10 μm, mobile phase: n-hexane/EtOH/NH3· _H2O = 60/40/0.1 (V/V/V), flow rate: 60 ml _/ min], and the separated solution was concentrated.

Stereomer 80-1 (peak 1) was obtained as a white solid (70 mg, t = 5.3 min). MS-ESI m/z: 490.2 [M+H]+. ^¹H NMR (400 MHz, DMSO-d6 ₎ )δ8.69–8.64(m,1H),8.44–8.39(m,1H),7.94(s,1H),7.87(d,J＝9.0Hz,1H),7.80–7.76(m,1H),7.74–7.69(m,1H),7.57(s,1H), 6.79(s,1H),6.43–6.35(m,3H),2.47–2.40(m,1H),2.33–2.22(m,2H),2.20–1.90(m,4H),1.89–1.65(m,4H),1.55–1.45(m,1H).

Stereomer 80-2 (peak 2) was obtained as a white solid (78 mg, t = 6.3 min). MS-ESI m/z: 490.2 [M+H]+. ^¹H NMR (400 MHz, DMSO-d6 ₎ )δ8.69–8.64(m,1H),8.44–8.39(m,1H),7.94(s,1H),7.87(d,J＝9.0Hz,1H),7.80–7.76(m,1H),7.74–7.69(m,1H),7.57(s,1H), 6.79(s,1H),6.43–6.35(m,3H),2.47–2.40(m,1H),2.33–2.22(m,2H),2.20–1.90(m,4H),1.89–1.65(m,4H),1.55–1.45(m,1H).

Examples 102 and 103:

Step 1.

Compound 69 (120 mg) was separated by chiral HPLC [Chiral ART cellulose-KCN (10 μm), 50 mm I.D. × 250 mm L (10 μm), mobile phase: n-hexane/EtOH = 90%/10% (V/V), flow rate: 100 ml/min], and the separated solution was concentrated.

Stereomer 69-1 (peak 1) was obtained as a white solid (69 mg, t = 25 min). MS-ESI m/z: 490.2 [M+H]+. ^1H NMR (400 MHz, DMSO- _d6) )δ8.70(d,J＝2.3Hz,1H),8.25(s,1H),8.08–8.02(m,1H),7.92–7.84(m,2H),7.77–7.71(m,1H),7.61(s,1H),6.90(s,2H ),6.82(s,1H),6.57(dd,J=7.7,4.8Hz,1H),2.35–2.25(m,2H),2.24–1.94(m,5H),1.93–1.70(m,4H),1.57–1.45(m,1H).

Stereomer 69-2 (peak 2) was obtained as a white solid (50 mg, t = 30 min). MS-ESI m/z: 490.2 [M+H]+. ^1H NMR (400 MHz, DMSO- _d6) )δ8.70(d,J＝2.3Hz,1H),8.25(s,1H),8.08–8.02(m,1H),7.92–7.84(m,2H),7.77–7.71(m,1H),7.61(s,1H),6.90(s,2H ),6.82(s,1H),6.57(dd,J=7.7,4.8Hz,1H),2.35–2.25(m,2H),2.24–1.94(m,5H),1.93–1.70(m,4H),1.57–1.45(m,1H).

Examples 104 and 105:

Step 1.

Compound 61 (460 mg) was separated by SFC [CHIRALPAK AD-H, mobile phase: _CO2 /EtOH = 80/20, flow rate: 90 ml/min], and the separated solution was concentrated.

Stereomer 61-1 (peak 1) was obtained as a white solid (205 mg, t = 9.2 min). MS-ESI m/z: 476.2 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO-d ₆ ) δ 8.39 (dd, J = 11.2, 2.8 Hz, 1H), 8.11 (s, 1H), 7.95 (dd, J = 9.2, 5.6 Hz, 1H), 7.70–7.58 (m, 2H), 7.38 (s, 1H), 6.79 (s, 1H), 3.75 (s, 3H), 2.29 (s, 3H), 2.29–1.68 (m, 11H), 1.58–1.42 (m, 1H).

Stereomer 61-2 (peak 2) was obtained as a white solid (199 mg, t = 13.2 min). MS-ESI m/z: 476.2 [M+H] ⁺ . ^1H NMR (400 MHz, DMSO-d ₆ ) δ 8.39 (dd, J = 11.2, 2.8 Hz, 1H), 8.11 (s, 1H), 7.95 (dd, J = 9.2, 5.6 Hz, 1H), 7.70–7.58 (m, 2H), 7.38 (s, 1H), 6.79 (s, 1H), 3.75 (s, 3H), 2.29 (s, 3H), 2.29–1.68 (m, 11H), 1.58–1.42 (m, 1H).

Example 106: Preparation of Compound 106

Step 1. Preparation of compound 106-1

2,4,6-Trichloroquinoline (200 mg) was dissolved in anhydrous dimethyl sulfoxide (5 mL), and (5-aminobicyclo[3.2.1]octane-1-yl)benzyl carbamate (236 mg) and N,N-diisopropylethylamine (445 mg) were added. The mixture was heated to 130 °C. After the reaction was completed, the mixture was cooled to room temperature, diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: petroleum ether-ethyl acetate = 5:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a yellow oily compound 106-1 (100 mg, yield 25%). MS-ESI m/z: 470.1 [M+H]+.

Step 2. Preparation of compound 106-2

Compound 106-1 (110 mg) was dissolved in TFA (1.8 mL) and water (0.2 mL), heated to 60 °C until the reaction was complete, concentrated, and the pH was adjusted to alkaline with ammonia. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated to give a yellow oily compound 106-2 (80 mg). The crude product was used directly in the next step. MS-ESI m/z: 336.1 [M+H]+.

Step 3. Preparation of compound 106

106-2 (80 mg) was dissolved in DMF (5 mL), and 2-aminonicotinic acid (39 mg), HATU (137 mg), and N,N-diisopropylethylamine (93 mg) were added. The reaction was carried out at room temperature. After the reaction was completed, the mixture was diluted with water, extracted with ethyl acetate, and the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: dichloromethane-methanol = 15:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a pale yellow solid compound 106 (5.1 mg, 5% yield in two steps). MS-ESI m/z: 456.1 [M+H]+. ¹ H NMR (400MHz, DMSO-d ₆ )δ8.58(d,J＝2.0Hz,1H),8.22(s,1H),8.07–8.02(m,1H),7.90–7.84(m,1H),7.73–7.62(m,2H),7.37(s,1H),6 .88(s,2H),6.57(dd,J＝7.6,4.8Hz,1H),6.51(s,1H),2.33–2.26(m,2H),2.20–2.03(m,4H),1.95–1.71(m,6H).

Example 107: Preparation of Compound 107

Step 1. Preparation of compound 107-1

100 mg of 4,6-dichloro-2-(difluoromethyl)quinoline was dissolved in 2 mL of anhydrous dimethyl sulfoxide. 110 mg of (5-aminobicyclo[3.2.1]octane-1-yl)carbamate and 210 mg of N,N-diisopropylethylamine were added, and the mixture was heated to 130 °C. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, extracted with ethyl acetate, and the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: petroleum ether-ethyl acetate = 5:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a yellow oily compound 107-1 (15 mg, yield 8%). MS-ESI m/z: 486.2 [M+H] ⁺ .

Step 2. Preparation of compound 107-2

Compound 107-1 (15 mg) was dissolved in a mixed solvent of TFA (1.8 mL) and water (0.2 mL) and heated to 60 °C. After the reaction was complete, the solvent was removed, the pH was adjusted to alkaline with ammonia, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and concentrated to give a yellow oily compound 107-2 (10 mg). The crude product was used directly in the next step. MS-ESI m/z: 352.1 [M+H] ⁺ .

Step 3. Preparation of Compound 107

Compound 107-2 (10 mg) was dissolved in DMF (2 mL), and 2-aminonicotinic acid (4.6 mg), HATU (16 mg), and N,N-diisopropylethylamine (10 mg) were added. The reaction was carried out at room temperature. After the reaction was completed, the solution was diluted with water, extracted with ethyl acetate, and the combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography [eluent: dichloromethane-methanol = 15:1]. The eluent was collected, and the solvent was removed under reduced pressure to give a white solid compound 107 (3.5 mg, two-step yield 24%). MS-ESI m/z: 472.2 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ )δ8.63(d,J＝2.3Hz,1H),8.23(s,1H),8.05(dd,J＝4.8,1.7Hz,1H),7.89–7.86(m,1H),7.84(d,J＝8.9Hz,1H),7.72–7.65(m, 1H),7.38(s,1H),6.91–6.75(m,4H),6.57(dd,J＝7.6,4.8Hz,1H),2.35–2.28(m,2H),2.24–2.07(m,4H),2.00–1.73(m,6H).

The following compounds were synthesized using a route similar to that in Example 107:

Biological tests

Experimental Example 1: Detection of Calcium Ion Concentration

Experimental materials:

HEK293T cells stably expressing human MRGPRX2 were constructed at Beijing Kanglong Pharmaceutical Technology Co., Ltd. Cells were cultured in a 37°C, 5% (v/v) _CO2 incubator in DMEM containing 10% FBS, 100 U/mL penicillin-streptomycin, 20 mM HEPES and 1 μg/mL puromycin.

Experimental methods:

1. HEK293T/MRGPRX2 cells were cultured in the above-mentioned medium until they reached the logarithmic growth phase. They were then digested with trypsin containing 0.25% EDTA, centrifuged at 1000 rpm, and the supernatant was discarded. Cell culture medium (5% FBS, 100 U/mL penicillin-streptomycin, 20 mM HEPES in DMEM) was added. After cell counting, the cell suspension was adjusted to 1 x ^10⁶ /mL. 20 μL of cell suspension per well was added to a 384-well cell culture plate and cultured overnight (culture conditions: 37℃, 5% (v/v) _CO₂ ).

2. Prepare the dye solution according to the instructions of the Screen Quest ^™ Fluo-8 No-Wash Calcium Assay Kit: Dilute component B (10x) with 90 mL of assay buffer (20 mM HEPES in 1x HBSS, pH 7.4). Prepare Fluo-8 loading buffer (F127 Plus, 10 mL). Add 200 μL DMSO to Fluo-8 NW reagent to obtain Fluo-8 NW stock solution. Add 200 μL of Fluo-8 NW stock solution to Fluo-8 loading buffer and vortex vigorously for 1-2 minutes to obtain dye solution. Add 20 μL of dye solution to each well of a 384-well cell culture plate and incubate at 25°C for 1 hour.

3. Preparation of the test compound and MRGPRX2 agonist (cortistatin-14): In the compound preparation plate, the 10 mM test compound was diluted 1000 times with DMSO to the final detection concentration, and then diluted 200 times with loading buffer to obtain a solution of the test compound at a final detection concentration of 5 times. The 10 mM cortistatin-14 was diluted 3.3 times with sterile water, and then serially diluted 1:3 (10 times). It was then diluted 200 times with loading buffer to obtain a solution of cortistatin-14 at a final detection concentration of 5 times.

4. Agonist Mode Data Detection: 10 μL of the above 5-fold corticostatin-14 solution was added to a 384-well cell culture plate using FLIPR, and fluorescence signal values were read at 160-second intervals. The EC90 value of corticostatin-14 was calculated using Graphpad Prism 9.0.

5. Antagonist Mode Data Detection: 10 μL of the above-mentioned 5x EC90 solution of the test compound was added to a 384-well cell culture plate using FLIPR. The 384-well cell culture plate was then incubated with the compound at room temperature in the dark for 30 minutes. 6x EC90 of corticostatin-14 was prepared using loading buffer. 10 μL of 6x EC90 corticostatin-14 was added to the 384-well cell culture plate using FLIPR. Fluorescence signal values were read at 1-second intervals after 160 seconds.

6. Analysis of the percentage inhibition rate of the test compound: % inhibition rate = (fluorescence value of positive control group - fluorescence value of compound group) / (fluorescence value of positive control group - fluorescence value of negative control group) * 100%, where the fluorescence value of the positive control group is the fluorescence value of the well with only agonist added, and the fluorescence value of the negative control group is the fluorescence value of the well without agonist added.

Experimental results: see Table 1.

Table 1: Calcium ion antagonistic activity

Experimental conclusion: The compounds in the examples of this application have strong antagonistic activity against MRGPRX2.

Experiment Example 2: Liver Microsomal Metabolism Experiment

Experimental materials: Mixed CD1 mouse liver microsomal enzyme protein was purchased from Xenotech (#M1000), and NADPH was purchased from Abmole (#M9076).

Experimental methods:

1. Add 210 μL of phosphate buffer and 12.5 μL of liver microsomal enzyme at a concentration of 20 mg/mL to the liver microsomal enzyme reaction system, then add 25 μL of NADPH at a concentration of 10 mM to the system, mix well, and then shake in a water bath at 37°C for 10 minutes.

2. Dilute the 10 mM sample of the test compound to 100 μM with DMSO, then add 2.5 μL of the diluted test compound to the reaction system, mix well, and then incubate in a water bath at 37 °C with shaking.

3. At 0.5, 5, 10, 20, 30, and 60 minutes, respectively, take 25 μL of the enzyme reaction solution and add it to 125 μL of cold acetonitrile containing dexamethasone as an internal standard. Centrifuge at 4000g for 20 minutes, and collect the supernatant and mix it with an equal volume of distilled water. Perform quantitative analysis of the mixed sample by liquid chromatography-mass spectrometry.

4. Calculation of in vitro drug metabolic half-life and drug clearance rate:

t1/2 = -0.693/k. (k: the slope of the linear regression of the natural logarithm of the remaining drug percentage with incubation time).

In vitro drug clearance rate = (0.693/t1/2) * (enzyme reaction volume / liver microsomal enzyme content).

The experimental results are shown in Table 2.

Table 2: Stability of the drug under mouse liver microsomal metabolism conditions

*EP4-400 is compound 4-400 reported in patent WO2022067094A1, and its structure is as follows:

Experimental conclusions: Compared with the positive control compound EP4-400, the compounds in Examples 7, 23, 61, 69, 80, and stereoisomer 80-2 of this application exhibit lower liver microsomal clearance rates, longer half-lives, and better liver microsomal metabolic stability.

Experiment Example 3: Hepatocyte clearance rate test

Experimental materials: Mouse hepatocytes (Bioreoclmation IVT, product catalog: M00505). Source: Male ICR/CD-1 mice.

Experimental methods:

1. Preparation of working solution

a. Prepare stock solutions of 10 mM test compound and positive control in DMSO.

b. In a separate conical tube, mix 198 μL of 50% acetonitrile/50% water and 2 μL of 10 mM solution to prepare a 100 μM test compound and positive control.

2. Preparation of hepatocytes

a. Place the incubation medium and hepatocyte thawing medium in a 37°C water bath and heat for at least 15 minutes before use.

b. Transfer a vial of cryopreserved liver cells from storage, ensuring the vial remains at a low temperature until thawing occurs. Thaw the cells by placing the vial in a 37°C water bath and gently shaking it for 2 minutes. Once thawed, spray the vial with 70% ethanol and transfer it to a biosafety cabinet.

c. Using a large-bore pipette tip, transfer the hepatocytes into a 50 mL conical tube containing thawed culture medium. Place the 50 mL conical tube in a centrifuge and centrifuge at 100 g for 10 minutes. After centrifugation, aspirate the thawed culture medium and resuspend the hepatocytes in sufficient culture medium to produce ~1.5 × ^10⁶ cells/mL.

3. Stability testing procedure

a. Transfer 198 μL of hepatocytes to each well of an uncoated 96-well plate (0.3 × ^10⁶ cells/well). Place the plate in an incubator to warm the hepatocytes for 10 minutes.

b. Pipe 2 μL of the 100 μM test compound or positive control into each well of the 96-well uncoated plate to begin the reaction. Return the plates to the incubator at the set time points.

c. Transfer the contents of the wells in 25 μL aliquots at time points of 0, 15, 30, 60, 90, and 120 minutes. Then, terminate the reaction by mixing the aliquot with 6 volumes (150 μL) of acetonitrile containing the internal standard IS (100 nM alprazolam, 200 nM caffeine, and 100 nM tolbutamide). Vortex for 5 minutes. Centrifuge the sample at 3220 g for 45 minutes. Dilute 100 μL of the supernatant aliquot with 100 μL of ultrapure water and use the mixture for LC/MS/MS analysis. All incubations were performed in duplicate.

d. Data Analysis

All calculations were performed using Microsoft Excel. Peak areas were determined from the extracted ion chromatograms. The in vitro half-life (t1/2) of the parent compound was determined by regression analysis of the percentage of compound disappearance against the time curve.

The in vitro half-life (t1/2) is determined by the slope value k:

t1/2 = 0.693/k

The in vitro t1/2 (in minutes) was converted to the in vitro intrinsic clearance (CLint, in μL/min/0.3× ^10⁶ cells) using the following equation (mean of repeated measurements):

CLint = kV/N

V = Incubation volume (0.2 mL)

N = Number of hepatocytes per well (0.3 × ^10⁶ cells)

Experimental conclusion: Stereoisomer 80-2 of the present application has good hepatocyte metabolic stability.

Experiment Example 4: Evaluation of Pharmacokinetics in Mice

Experimental Objective: To test the pharmacokinetics of the compounds in the embodiments of this application in mice. By detecting the drug concentration in mouse plasma, the pharmacokinetic behavior of the compounds of this invention in mice is studied, and their pharmacokinetic characteristics are evaluated.

Test animals: 6-8 week old male ICR (CD1) mice

Experimental methods:

Preparation of oral gavage medication: First, weigh an appropriate amount of the test compound and dissolve it in DMSO (Macklin CAS: 67-68-5) to prepare a 20 mg/mL solution. Take 75 μL of the above solution and add it to 150 μL of Solutol (Sigma CAS: 70142-34-6), then add 1.275 mL of 20% Captisol (Selleck CAS: 182410-00-0) to finally obtain a 1 mg/mL dose of the drug.

Preparation of intravenous injection drug: First, weigh an appropriate amount of the test compound and dissolve it in DMSO (Macklin CAS: 67-68-5) to prepare a 30 mg/mL solution. Take 15 μL of the above solution and add it to 150 μL of Solutol (Sigma CAS: 70142-34-6), then add 1.335 mL of 20% Captisol (Selleck CAS: 182410-00-0) to finally obtain a 0.3 mg/mL drug.

Specific procedures: Mice were fasted overnight but allowed free access to water before the experiment. Feeding was resumed two hours after drug administration. Blood samples were collected at different time points after intravenous injection or oral gavage of the test compound to determine the concentration of the compound in the plasma. Approximately 50 μL of blood was collected from each animal via a capillary sampling tube through the orbital vein, and heparin sodium was used for anticoagulation. After collection, the blood samples were placed on ice and centrifuged at 3500 rpm for 10 min. The plasma was separated, and 20 μL of the collected plasma was added to 200 μL of acetonitrile (Merck CAS: 75-05-8) containing 200 nM dexamethasone (Selleck CAS: 50-02-2) as an internal standard, followed by centrifugation at 8000 rpm for 20 min. After centrifugation, 150 μL of the supernatant was transferred to a new centrifuge tube, and 150 μL of 0.1% formic acid (Fisher CAS: 207868) was added. After mixing, take 5 μL of the sample for quantitative detection of compounds by liquid chromatography-mass spectrometry (Q-TOF LC/MS).

Standard curve determination:

1. The test compound was serially diluted with DMSO to a concentration that covered the concentration of the compound in the plasma to be tested, and a blank sample (containing only DMSO) was prepared.

2. Take 2 μL of each of the above diluted samples and add them to 18 μL of normal ICR mouse plasma. Mix well, then add 200 μL of acetonitrile (Merck CAS: 75-05-8) containing 200 nM dexamethasone (Selleck CAS: 50-02-2) as an internal standard. Centrifuge for 20 min (8000 r/min). Transfer 150 μL of the supernatant to a new centrifuge tube, add 150 μL of 0.1% formic acid (Fisher CAS: 207868), mix well, and then take 5 μL of the sample for quantitative analysis of compounds by liquid chromatography-mass spectrometry (Q-TOF LC/MS).

3. Finally, with the concentration of each diluted test compound as the x-axis and the ratio of the obtained signal of each compound to the internal standard (dexamethasone) as the y-axis, a standard curve was generated using the linear regression method (R2>0.9900) in GraphPad Prism 8 software.

Data Analysis: Calculation of Pharmacokinetic Parameters: Based on the above standard curve, the plasma concentrations of the test compound at different time points after intravenous injection or oral gavage in ICR mice were calculated. Pharmacokinetic parameters (T1/2, Tmax, Cmax, AUC, etc.) were calculated using the Phoenix WinNonlin 8.1 non-compartmental analysis model.

Experimental results: see Table 3.

Table 3: Pharmacokinetic data of mice by gavage (PO, 10 mpk)

Experimental conclusions: Compared with the positive control compound EP4-400, the compounds in Examples 35, 7, 69, 80, stereoisomer 80-2, and stereoisomer 61-1 of this application exhibit higher drug exposure, higher peak concentration, and longer in vivo half-life in mice, demonstrating better pharmacokinetic properties and drug-likeness.

Experimental Example 5: Pharmacokinetic Evaluation in Rats

Experimental Objective: To test the pharmacokinetics of the compounds in the embodiments of this application in rats. By detecting the drug concentration in rat plasma, the pharmacokinetic behavior of the compounds of this invention in rats is studied, and their pharmacokinetic characteristics are evaluated.

Test animals: 6-8 week old male SD rats

Experimental methods:

Preparation of oral gavage medication: First, weigh an appropriate amount of the test compound and dissolve it in DMSO (Macklin CAS: 67-68-5) to prepare a 100 mg/mL solution. Take 100 μL of the above solution and add it to 200 μL of Solutol (Sigma CAS: 70142-34-6), then add 1.7 mL of 20% Captisol (Selleck CAS: 182410-00-0) to finally obtain a 5 mg/mL dose of the drug.

Preparation of intravenous injection drug: First, weigh an appropriate amount of the test compound and dissolve it in DMSO (Macklin CAS: 67-68-5) to prepare a 60 mg/mL solution. Take 50 μL of the above solution and add it to 100 μL of Solutol (Sigma CAS: 70142-34-6), then add 850 μL of 20% Captisol (Selleck CAS: 182410-00-0) to finally obtain a 3 mg/mL drug.

Specific procedures: Rats were fasted overnight but allowed free access to water before the experiment. Feeding was resumed two hours after drug administration. Blood samples were collected at different time points after intravenous injection or oral gavage of the test compound to determine the concentration of the compound in the plasma. Approximately 50 μL of blood was collected from each animal via a capillary sampling tube through the orbital vein, with heparin sodium as anticoagulant. After collection, the blood samples were placed on ice and centrifuged at 3500 rpm for 10 min to separate the plasma. 20 μL of the collected plasma was added to 200 μL of acetonitrile (Merck CAS: 75-05-8) containing 200 nM dexamethasone (Selleck CAS: 50-02-2) as an internal standard, followed by centrifugation at 8000 rpm for 20 min. 150 μL of the supernatant was transferred to a new centrifuge tube, and 150 μL of 0.1% formic acid (Fisher CAS: 207868) was added. After mixing, 5 μL of the sample was used for quantitative analysis of the compound using liquid chromatography-mass spectrometry (Q-TOF LC/MS).

Standard curve determination:

1. The test compound was serially diluted with DMSO to a concentration that covered the concentration of the compound in the plasma to be tested, and a blank sample (containing only DMSO) was prepared.

2. Take 2 μL of each of the above diluted samples at different concentrations and add them to 18 μL of normal SD rat plasma. Mix well, then add 200 μL of acetonitrile (Merck CAS: 75-05-8) containing 200 nM dexamethasone (Selleck CAS: 50-02-2) as an internal standard. Centrifuge for 20 min (8000 r/min). Transfer 150 μL of the supernatant to a new centrifuge tube, add 150 μL of 0.1% formic acid (Fisher CAS: 207868), mix well, and then take 5 μL of the sample for quantitative analysis of compounds by liquid chromatography-mass spectrometry (Q-TOF LC/MS).

3. Finally, with the concentration of each diluted test compound as the x-axis and the ratio of the obtained signal of each compound to the internal standard (dexamethasone) as the y-axis, a standard curve was generated using the linear regression method (R2>0.9900) in GraphPad Prism 8 software.

Data analysis and calculation of pharmacokinetic parameters: The plasma concentrations of the test compound at different time points after intravenous injection or oral gavage in SD rats were calculated based on the above standard curve. Pharmacokinetic parameters (T1/2, Tmax, Cmax, AUC, etc.) were calculated using the Phoenix WinNonlin 8.1 non-compartmental analysis model.

Experimental results: see Table 4.

Table 4: Pharmacokinetic data of rats administered via gavage (PO, 10 mpk)

Experimental conclusion: Stereoisomer 80-2 of this application, compared with the positive control compound EP4-400, exhibits higher drug exposure, higher peak drug concentration and longer in vivo half-life in rats, demonstrating better pharmacokinetic properties and drug-likeness.

In addition to those described herein, various modifications of the invention will be apparent to those skilled in the art based on the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. All references cited in this application (including all patents, patent applications, journal articles, books, and any other disclosures) are incorporated herein by reference in their entirety.

Claims (14)

A compound or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound has the structure of formula (I):
in: Ring A, ring B, and ring D are each independently a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring; The ring C is a _C3-6 hydrocarbon ring or a 3-10 membered heterocycle; ^R1 , ^R2 , ^R5 , and ^R6 are each independently selected from halogens, -OH, _-NH2 , -CN, _-NO2 , _-SF5 , = _CH2 , -CH ^{= CRaRb} , _C1-6 alkyl, deuterated _C1-6 alkyl, halo- _C1-6 alkyl, _C2-6 alkenyl, C2-6 ynyl, _C3-6 cyclic hydrocarbon, _3-10 membered heterocyclic, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Ra , -OC(=O ^{) Ra} , -C(=O) ^ORa _, ^-ORa , -SRa, -S(=O) ^Ra , -S(= ^O ) _2Ra , ^-S (=O) ^2NRaRb , -S(=O)(= ^NRa ) ^Rb -NR ^a R ^b , -C(＝O)NR ^a R ^b , -C(＝O)N(R ^a )-OR ^b , -NR ^a -C(＝O)R ^b , -NR ^a -C(＝O)OR ^b , -NR ^a -S(＝O) ₂ -R ^b , -NR ^a -C(＝O)-NR ^a R ^b , -N＝S(＝O)R ^a R ^b , -P(＝O)R ^a R ^b , -C _1-6 alkylene-R ^a , -C _1-6 alkylene-OR ^a , -C _1-6 alkylene-NR ^a R ^b , -OC _1-6 alkylene-NR ^a R ^b , (-C _3-6 cycloene)-CN and (-C _3-6 cycloene)-C _1-6 alkyl; Alternatively, when p≥2, two ^R5s on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring. Alternatively, when q≥2, two ^R6s on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a _C6-10 aromatic ring, or a 5-14 membered heteroaromatic ring. ^R3 and ^R4 are each independently selected ^from H, D, -OH, _-NH2 , -CN, -CH= ^CRaRb , _C1-6 alkyl, deuterated _C1-6 alkyl, halogenated _C1-6 alkyl, C2-6 alkenyl, C2-6 ynyl, _C3-6 cycloalkyl, _3-10 heterocyclic, _C6-10 aryl, _5-14 heteroaryl, _C6-12 aralkyl, -C(=O ^{) Ra} , -C(=O) ^ORa , -S ⁽ =O) ^Ra , -S(=O) _2Ra , -S(=O) ^2NRaRb , -S(=O)(= ^{NRa )Rb ,} -C(=O) ^NRaRb , -C(= _O )N( ^Ra ) ^-ORb , _-C1-6 alkylene- ^Ra , -C _1-6 alkylene-OR ^a , -C _1-6 alkylene-NR ^a R ^b , -OC _1-6 alkylene-NR ^a R ^b , (-C _3-6 cycloene)-CN and (-C _3-6 cycloene)-C _1-6 alkyl; ^Ra and ^Rb are each independently selected from H, _C1-6 alkyl, C3-10 cyclic hydrocarbon, _3-10 membered heterocyclic group, _C6-10 aryl, 5-14 membered heteroaryl, and _C6-12 aryl group each time they appear; or ^Ra and ^Rb together with the groups they are attached to form a _C3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a _C6-10 aromatic ring, or a 5-14 membered heteroaryl ring. The aforementioned alkylene, alkyl, alkenyl, alkynyl, cycloalkylene, cycloalkylene, cycloalkylene, heterocyclic, heterocyclic, aryl, aromatic, heteroaryl, heteroaromatic, and aralkyl groups are each optionally substituted by one or more substituents independently selected from the following: deuterium, halogen, -OH, =O, _-NH₂ , -CN, _-NO₂ , = _CH₂ , = _CF₂ , -CH= ^CRcRd , _C1-6 alkyl, deuterated _C1-6 alkyl, _C2-6 alkenyl, _C2-6 ^alkynyl , C3-6 cycloalkyl, _3-10 membered heterocyclic, _C6-10 aryl, 5-14 membered heteroaryl, _C6-12 aralkyl, -C(=O) ^Rc , -OC(=O) ^Rc , -C(=O) ^ORc , ^-ORc , ^-SRc , -S(=O) ^Rc -S(=O) _₂R₁c , -S(=O ^{) ₂NR₁cR₂d} , -NR₁cR₂d ^{, -C} (=O ^{) NR₁cR₂d} , -NR₁c ^{- C} (=O) ^R₂d , -NR₁c ^- _C (=O) ^OR₂d , -NR₁c ^- S(=O) _₂- ^R₂d , -NR₁c ^-C (=O ^{) -NR₁cR₂d} , -N=S(=O) ^R₁cR₂d , -C₁ _- 6alkylene- ^OR₁c , -C₁- _6alkylene ^{- NR₁cR₂d} and -OC₁- _6alkylene ^{- NR₁cR₂d} , wherein the alkylene, alkyl, alkenyl, = _CH₂ , alkynyl, ^cycloalkyl , heterocyclic, aryl, heteroaryl and aralkyl groups are each optionally further substituted by one or more substituents independently selected from: halogen, -OH, =O, -C(=O)O-tert-butyl, -NH ₂ , -CN, _-NO2 , = _CH2 , = _CF2 , _C1-6 alkyl, _C1-6 haloalkyl, _C3-6 cyclic hydrocarbon, 3-10 heterocyclic, _C6-10 aryl, 5-14 heteroaryl, _C6-12 aralkyl, _-C1-6 alkylene- _C3-6 cyclic hydrocarbon, _-OC1-6 alkyl and _-C1-6 alkylene- _OC1-6 alkyl; ^Rc and ^Rd, each in each occurrence, are independently selected from H, _C1-6 alkyl, C3-10 cyclic hydrocarbon, _3-10 membered heterocyclic group, _C6-10 aryl, 5-14 membered heteroaryl, and _C6-12 aralkyl, or ^Rc and ^Rd together with the groups they are attached to constitute a C3-6 hydrocarbon ring, a 3-10 membered heterocyclic ring, a _C6-10 aromatic ring, or a 5-14 membered heteroaryl ring. The alkyl, _cyclic hydrocarbon, hydrocarbon ring, heterocyclic group, heterocyclic ring, aryl, aromatic ring, heteroaryl, heteroaryl, and aralkyl groups are further optionally substituted by one or more substituents independently selected from: halogen, -OH, =O, -C(=O)O-tert-butyl, _-NH₂ , -CN, _-NO₂ , _C1-6 alkyl, _C1-6 haloalkyl, C3-6 _cyclic hydrocarbon, 3-10 membered heterocyclic group, C _6-10 aryl, 5-14 heteroaryl, _C6-12 aralkyl and _-C1-6 alkylene- _OC1-6 alkyl; m, n, p, and q are each independent integers selected from 0, 1, 2, 3, and 4; The condition is 1) when for and for When p is not 0 or m ≥ 2; 2) When for and for If p is not 0 or at least one ^R2 is _-SF5 . The compound of claim 1 or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ring A and ring B are each independently _{a C6-10} aromatic ring or a 5-14 membered heteroaromatic ring. Preferably, ring A and ring B are each independently a benzene ring or a 5-6 membered heteroaromatic ring; More preferably, Selected from The compound of claim 1 or 2 or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R1 and ^R2 are each independently selected from halogen, -CN, _-SF5 , _C1-6 alkyl, halogenated _C1-6 alkyl and -O-( _C1-6 alkyl) in each occurrence; Preferably, ^R1 and ^R2 are each independently selected from F, Cl, CN, _-SF5 , _CHF2 , _CF3 and _-OCH3 each time they appear. The compound of any one of claims 1-3 or the pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R3 and ^R4 are both H. The compound of any one of claims 1-4, or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein when for and for When p≥2, the two ^R5s located on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-10 membered heterocycle, a benzene ring, or a 5-14 membered heteroaromatic ring; preferably, at this time, for The compound of any one of claims 1-5 or the pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R5 is independently _C1-6 alkyl in each occurrence; or When p≥2, two ^R5 atoms on the same or different ring atoms, together with the groups they are attached to, constitute a _C3-6 hydrocarbon ring, a 3-6 membered heterocycle, a benzene ring, or a 5-6 membered heteroaromatic ring; or When p≥2, two ^R5 atoms located on the same or different ring atoms together constitute a _C1-4 alkylene, preferably a _C1-3 alkylene, and more preferably an ethylene; or p=2, the two ^R5s are located on different ring atoms, and the two ^R5s together constitute a _C1-4 alkylene, preferably a _C1-3 alkylene, and more preferably an ethylene. The compound of any one of claims 1-6, or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein... Selected from The compound of any one of claims 1-7 or the pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ring D is selected from 5-6 membered heterocycles, benzene rings or 5-6 membered heteroaromatic rings; preferably, ring D is selected from benzene rings or 5-6 membered heteroaromatic rings. The compound of any one of claims 1-8 or the pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein ^R6 is independently a halogen, _-NH2 , -CN, _C1-6 alkyl, halo _-C1-6 alkyl, _C3-6 cycloalkyl, phenyl, -C(=O)-( _C1-6 alkyl), -O-( _C1-6 alkyl), -NH( _C1-6 alkyl), -NH(halo- _C1-6 alkyl), -NH _- S(=O) _2- ( _C1-6 alkyl), _-C1-6 alkylene-CN or _-C1-6 alkylene-OH; Alternatively, when q≥2, two ^R6 atoms on the same or different ring atoms, together with the groups they are attached to, constitute a benzene ring or a 5-6 membered heteroaromatic ring. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound, or prodrug thereof, wherein... Selected from The compound of any one of claims 1-10, or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound has the structure of formula (II), (III) or (IV):
in p’ is 0, 1 or 2; The remaining groups are as defined in any one of claims 1-10. The compound of any one of claims 1-11 or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug thereof, wherein the compound is selected from:









A pharmaceutical composition comprising a preventive or therapeutically effective amount of the compound of any one of claims 1-12 or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug, and one or more pharmaceutically acceptable carriers. The use of any compound of claims 1-12 or a pharmaceutically acceptable salt, ester, N-oxide, stereoisomer, tautomer, polymorph, solvate, metabolite, isotopically labeled compound or prodrug, or the pharmaceutical composition of claim 13, in the preparation of a medicament used as an MRGPRX2 antagonist; preferably, the medicament is used to prevent or treat diseases that can be improved by antagonizing MRGPRX2, preferably, the diseases are selected from urticaria, drug-induced acute allergic/anaphylactic reactions, asthma, dermatitis (e.g., atopic dermatitis), pruritus, allergies (e.g., food allergies), enteritis, irritable bowel syndrome, arthritis, allergic rhinitis, nasal polyps, and other diseases related to type II inflammatory response and mast cell-related diseases.