WO2022117062A1

WO2022117062A1 - Fused tricyclic ring-containing compounds and pharmaceutical use thereof

Info

Publication number: WO2022117062A1
Application number: PCT/CN2021/135223
Authority: WO
Inventors: 李文明; 刘宁; 邹昊; 李云飞; 张芳; 张瑱
Original assignee: 上海拓界生物医药科技有限公司
Priority date: 2020-12-04
Filing date: 2021-12-03
Publication date: 2022-06-09
Also published as: TW202227458A

Abstract

The present disclosure relates to fused tricyclic ring-containing compounds and the pharmaceutical use thereof. Such compounds can be used for treating P2X3 activity-related diseases. Exemplary compounds are as represented by formula I, and the definitions of the groups are as described in the description.

Description

Compound containing condensed tricyclic ring and its medicinal use

technical field

The present disclosure relates to the field of medicine, in particular to a class of compounds containing fused tricyclic rings and their medicinal uses.

Background technique

P2X receptors are a family of cation-permeable ligand-gated ion channels that open in response to the binding of extracellular adenosine 5'-triphosphate (ATP). They belong to a larger family of receptors called purinergic receptors. P2X receptors are present in a variety of organisms, including humans, mice, rats, rabbits, chickens, zebrafish, bullfrogs, flukes, and amoeba. Seven independent genes encoding P2X subunits have been identified and named P2X1 to P2X7. Different subunits exhibit different sensitivities to purinergic agonists and antagonists.

P2X3 receptors have 4 ATP binding sites on a single subunit, consisting of 2 transmembrane domains, N-terminal and C-terminal located in the cell, and a conserved sequence located in the extracellular loop structure. The high expression of P2X3 receptors was found in specific small and medium diameter neurons related to nociceptive information. At the same time, P2X3 receptors are also involved in the transmission of some non-nociceptive sensations. It has been confirmed that the P2X3 receptor is involved in bladder sensory function and is a key receptor-mediated bladder sensory signal, expressed in the bladder mucosal tissue rich in sensory nerve fibers. P2X3 is also expressed in the sensory nerve fibers of the pharyngeal mucosa, which is related to the conduction and formation of taste.

When the body is injured or nerve damage, a large amount of ATP is released, which activates P2X3 receptors in the presynaptic membrane, causing a large influx of Ca2+, and the increase in intracellular calcium concentration activates protein kinase A (PKA) and protein kinase C (protein kinase C). C, PKC), which phosphorylates PKA and PKC, and at the same time promotes the release of glutamate, which further activates NMDA receptors, resulting in the generation of excitatory postsynaptic currents and central sensitization. Many studies have shown that up-regulation of P2X3 receptor expression can lead to the formation of pain sensitivity and participate in pain signaling.

MK-7264 is a P2X3 receptor activity antagonist, its IC50 values for human homologous recombination hP2X3 and hP2X2/3 are ~30nM and 100-250nM, respectively, and its use in the treatment of chronic cough patients has been carried out to clinical III Expect.

SUMMARY OF THE INVENTION

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

Wherein, ring A and ring B are independently selected from 6-membered aromatic ring, 5-7-membered heteroaromatic ring and 5-7-membered heterocyclic ring, and ring A and ring B are connected by sharing 2 atoms;

Y is

Wherein, ring C is a 5-8 membered heterocyclic ring or a 5-8 membered heteroaromatic ring, and ring D is a 5-8 membered heterocyclic ring or a 5-8 membered heteroaromatic ring, and ring C and ring D share a nitrogen atom and 1 carbon atom is connected, and ring D and the benzene ring are connected by sharing 2 carbon atoms;

Q is

R ₃ is independently selected from hydrogen, deuterium, halogen, C _1-4 alkyl, C _3-8 cycloalkyl and C _3-8 cycloalkylene, the C _1-4 alkyl, C _3-8 ring Alkyl and C _3-8 cycloalkane are optionally substituted with one or more deuterium or halogen; alternatively, R on adjacent carbon atoms forms a C _3- optionally substituted with one or more _deuterium or halogen ₈ cycloalkanes;

R ₄ is selected from hydrogen, deuterium and the following groups optionally substituted with one or more substituents: C _1-6 alkyl, C _1-6 alkoxy, C _3-8 cycloalkyl and C _3-8 cycloalkylmethylene, the substituents being selected from deuterium, halogen, -OH and -CN;

X is selected from -O-, -CR _a1 R _a2 - and -NR _b -;

R _5a and R _5b are independently selected from hydrogen, deuterium, halogen and C _1-6 alkyl optionally substituted with deuterium or halogen;

R _a1 and R _a2 are independently selected from hydrogen, deuterium, halogen, C _1-6 alkyl optionally substituted with one or more deuterium or halogen, and C _3-8 optionally substituted with one or more deuterium or halogen Cycloalkyl; or, R _a1 , R _a2 and the carbon atoms to which they are commonly attached form a C _3-6 cycloalkylene optionally substituted by one or more deuterium or halogen;

R _b is selected from hydrogen, deuterium and the following groups optionally substituted with one or more substituents: C _1-6 alkyl, -C(=O)-C _1-4 alkyl and -S(=O) ₂ -C _1-4 alkyl, the substituent is selected from: deuterium, halogen, -OH and -CN;

m is an integer selected from 0 to 9, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9;

n is an integer selected from 0 to 7, for example, 0, 1, 2, 3, 4, 5, 6, and 7.

In some embodiments, the compound of formula I is:

Wherein, Z ₁ is selected from C, CR', O, N and NR", Z ₂ , Z ₃ , Z ₄ , Z ₅ , Z ₆ and Z ₇ are independently C or N, and ring A and ring B are aromatic ;

R ₁ is selected from the group consisting of hydrogen, deuterium, halogen and the following groups optionally substituted with one or more substituents: C _1-6 alkyl, C _1-6 alkoxy, C _3-8 cycloalkyl and C _{3 -8} cycloalkylmethylene, the substituent is selected from: deuterium, halogen, -OH and -CN;

R ₂ is independently selected from hydrogen, deuterium, halogen, C _1-6 _alkyl and C _3-8 cycloalkyl, which _are optionally replaced by one or more substituted by deuterium, halogen or -OH;

R' is selected from hydrogen, deuterium, halogen and C _1-4 alkyl optionally substituted with one or more deuterium or halogen;

R" is selected from hydrogen, deuterium, C ₁ -C ₆ alkyl and C _3-8 cycloalkyl, said C ₁ -C ₄ alkyl and C _3-8 cycloalkyl optionally being substituted by one or more deuterium or halogen substitution;

m is an integer selected from 0 to 3; the groups Y and Q are as defined for the compound of formula I.

In some embodiments, Z ₁ is N or O; Z ₁ is preferably N.

In some embodiments, in the compound of formula I or formula I' or a pharmaceutically acceptable salt thereof, the compound represented by formula I is selected from:

R ₁ , R ₂ , m, Y, Q are as defined for compounds of formula I'.

In some embodiments, the compound of formula I is selected from formula II-a, formula II-b, formula II-c, formula II-d, formula II-e, formula II-f, formula II-g, formula II -h, compounds of formula II-i and II-j; preferably compounds selected from II-a, formula II-b, formula II-c, formula II-d, formula II-e, formula II-f and formula II-g ; more preferably from compounds of formula II-a, formula II-b, formula II-e, formula II-f and formula II-g; even more preferably from compounds of formula II-a, formula II-b and formula II-g.

In one embodiment, the compound of formula I is

In another embodiment, the compound of formula I is

In some embodiments, in a compound of Formula I, Formula I', or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, Ring C is

And ring C is a 5-8 membered heterocycle; preferably ring C is

In some embodiments, Ring C is selected from

R ₆ is independently selected from hydrogen, deuterium, halogen, C _1-6 alkyl and C _3-6 cycloalkylene, said C _1-6 alkyl and C _3-6 cycloalkylene are optionally replaced by one or Multiple deuterium or halogen substitution; alternatively, R _on adjacent carbon atoms forms C _cycloalkane optionally substituted with one or more deuterium or halogen;

p is an integer selected from 1-3, and q is an integer selected from 0-9.

In some embodiments, Ring C is

In some embodiments, Ring C is

preferably

_In some embodiments, the substituents R of ring C are independently selected from hydrogen, deuterium, halogen, and _C1-4 alkyl optionally substituted with one or more deuterium or halogen _; preferably R is independently selected from hydrogen, deuterium and halogen _; more preferably R6 is independently hydrogen or deuterium.

In one embodiment, Ring C is

In other embodiments, in a compound of Formula I, Formula I', or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, Ring C is a 5-membered heteroaromatic ring.

In some embodiments, Ring C is

_ci , c2 and c3 are each independently CR' or N, where _R ' is as defined for compounds of _formula I'.

In some embodiments, Ring C is

c ₁ and c ₂ are as previously defined.

While in some embodiments, Ring C is selected from

preferably

In other embodiments, in compounds of Formula I, Formula I' or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, Ring D is selected from

R ₇ is independently selected from hydrogen, deuterium, halogen, C _1-6 alkyl, C _3-6 cycloalkyl and C _3-6 cycloalkylene, the C _1-6 alkyl, C _3-6 ring Alkyl and _{C3-6cycloalkylene} are optionally substituted with one or more deuterium or halogen; alternatively, _R7 _on adjacent carbon atoms forms C3 optionally substituted with one or more deuterium or halogen _-6 cycloalkanes,

r is an integer selected from 0-3; s is an integer selected from 0-7.

In some embodiments, Ring D is selected from

In some embodiments, Ring D is selected from

In other embodiments, in compounds of Formula I, Formula I' or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, Y is selected from:

In yet other embodiments, in compounds of Formula I, Formula I', or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, _R5a and _R5b are independently selected from hydrogen, deuterium, and halogen.

In some embodiments, R _5a and R _5b are independently selected from hydrogen, fluoro and chloro; preferably both R _5a and R _5b are fluoro.

In other embodiments, in a compound of Formula I' or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, R ₁ is selected from hydrogen, deuterium, halogen, and optionally substituted with one or more substituents The following groups: C _1-6 alkyl and C _3-8 cycloalkylmethylene, the substituents being deuterium or halogen.

In some embodiments, R ₁ is selected from hydrogen, deuterium, halogen, and C _1-6 alkyl optionally substituted with one or more deuterium or halogen; R ₁ is preferably deuterium optionally substituted with one or more or Halogen substituted methyl.

In other embodiments, in compounds of Formula I' or Formula II-a to Formula II-n, or pharmaceutically acceptable salts thereof, R ₂ is independently selected from hydrogen, deuterium, halogen, and C _1-6 alkyl, the _C1-6 alkyl is optionally substituted with one or more deuterium or halogen; preferably R2 is independently selected from _hydrogen , deuterium and halogen.

In other embodiments, in a compound of Formula I, Formula I' or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, Q is

In some embodiments, in a compound of Formula I, Formula I', or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, _R3 is independently selected from hydrogen, deuterium, halogen, _C1-4 alkyl, and C _3-8 cycloalkylene, the C _1-4 alkyl and C _3-8 cycloalkylene are optionally substituted by one or more deuterium or halogen; or, _R on adjacent carbon atoms forms _C3-8 cycloalkane optionally substituted with one or more deuterium or halogen.

In some embodiments, R ₃ is independently selected from hydrogen, deuterium, halo, and C _1-4 alkyl optionally _substituted with one or more deuterium or halo; or, adjacent _R on a carbon atom forms a C _cycloalkane optionally substituted with one or more deuterium or halogen;

Preferably _R3 is independently selected from hydrogen, deuterium and halogen.

In other embodiments, in a compound of Formula I, Formula I' or Formula II-a to Formula II _- n, or a pharmaceutically acceptable salt thereof, R4 is selected from hydrogen, deuterium and optionally substituted with one or more substituents of the following groups: C _1-6 alkyl, C _1-6 alkoxy and C _3-8 cycloalkyl, and the substituents are deuterium or halogen.

In some embodiments, R ₄ is the following group C _1-4 alkyl or C _1-4 alkoxy optionally substituted with one or more substituents that are deuterium or halo.

In some embodiments, R ₄ is methyl or ethyl; R ₄ is preferably methyl.

_In one embodiment, R4 is methoxy.

In some embodiments, in a compound of Formula I, Formula I' or Formula II-a to Formula II-n, or a pharmaceutically acceptable salt thereof, X is selected from -O-, -CR _a1 R _a2 - and -NR _b -;

R _a1 and R _a2 are independently selected from hydrogen, deuterium, halogen and C _1-6 alkyl optionally substituted with one or more deuterium or halogen; alternatively, R _a1 , R _a2 and the carbon atom to which they are commonly attached form an optional C _3-6 cycloalkylene substituted with one or more deuterium or halogen;

R _b is selected from hydrogen, deuterium and C _1-6 alkyl optionally substituted with one or more deuterium or halogen.

In one embodiment, wherein X is O.

In a second aspect, the present disclosure also provides a series of compounds or pharmaceutically acceptable salts thereof, selected from:

In the third aspect, the present disclosure also provides a method for preparing the compound described in the first or second aspect or a pharmaceutically acceptable salt thereof. For example, the synthesis of the compound of formula II-a includes the following steps:

The compound of formula III-a and the compound of formula III-b are subjected to nucleophilic substitution reaction under basic conditions to obtain the compound of formula III-c; the compound of formula III-c and the compound of formula III-d are subjected to reduction ring closure reaction to obtain formula II- a compound; R ₁ , R ₂ , R ₃ , R ₄ , X, Y, m, n are as defined in the first aspect, X ₁ is selected from halogen, sulfonyl and sulfinyl.

The synthesis of the compound of formula II-a can also include the following steps:

The compound of formula III-a and the compound of formula III-b are subjected to nucleophilic substitution reaction under basic conditions to obtain the compound of formula III-c; the compound of formula III-c and the compound of formula III-e are subjected to reduction ring-closing reaction to obtain the compound of formula III- Compound f; compound of formula III-f is subjected to CC coupling reaction under basic conditions to obtain compound of formula II-a; R ₁ , R ₂ , R ₃ , R ₄ , X, Y, m, n are as in the first aspect As defined, X ₁ , X ₂ are selected from halogen, sulfonyl and sulfinyl.

In other embodiments, the synthesis of compounds of formula II-b comprises the steps of:

The compound of formula III-g and the compound shown in formula III-h and formula III-d undergo cyclization reaction under catalyst conditions to obtain the compound shown in the formula; the catalyst is selected from palladium/carbon, Raney nickel, tetra- Phenylphosphine palladium, palladium dichloride, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]dichloride palladium, 1,1'-bis(dibenzylphosphorus)dichloride Dipentyl iron palladium, tris(dibenzylideneacetone)dipalladium or 2-biscyclohexylphosphine-2',6'-dimethoxybiphenyl, [1,1'-bis(di-tert-butylphosphino) ) ferrocene]dichloropalladium(II), cuprous iodide, cuprous bromide, cuprous chloride, copper triflate; R ₁ , R ₂ , R ₃ , R ₄ , X, Y , m, n are as defined in the first aspect.

In a fourth aspect, the present disclosure also provides a pharmaceutical composition comprising the compound described in the first or second aspect or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

In certain embodiments, the pharmaceutical composition contains 0.01-99.99% of the aforementioned compound or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1-99.9% of the aforementioned compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of the compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 1%-99% of the compound or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition contains 2%-98% of the compound or a pharmaceutically acceptable salt thereof.

In the fifth aspect, the present disclosure also provides the compound of the first or second aspect or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition comprising the same as described in the fourth aspect, in the preparation of a medicament for the treatment of a disease associated with P2X3 activity use. In some embodiments, the P2X3 activity-related disease refers to a P2X3 overactivity-related disease. The compounds of the present disclosure are highly selective for P2X3 and can avoid taste loss. In some embodiments, compounds of the present disclosure are more than 20-fold more potent at P2X3 homologous receptor antagonism than at P2X2/3 heteromeric receptors. In some embodiments, the compounds of the present disclosure are more than 30-fold more potent at P2X3 homologous receptor antagonism than at P2X2/3 heteromeric receptors. In some embodiments, the compounds of the present disclosure are more than 50-fold more potent at P2X3 homologous receptor antagonism than at P2X2/3 heteromeric receptors. In some embodiments, the compounds of the present disclosure are more than 100-fold more potent at P2X3 homologous receptor antagonism than at P2X2/3 heteromeric receptors.

In some embodiments, a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, can be used for the treatment of pain, urinary tract disease, cough, and the like. Pain can be, for example, chronic pain, neuropathic pain, acute pain, back pain, cancer pain, pain due to rheumatoid arthritis, migraine, and visceral pain. Urinary tract disorders such as overactive bladder (also known as urinary incontinence), pelvic hypersensitivity, and urethritis.

In some embodiments, a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, may be used to treat gastrointestinal disorders, including, for example, constipation and functional gastrointestinal disorders (eg, irritable bowel syndrome or functional gastrointestinal disorders). indigestion); can be used in the treatment of cancer; can be used in the treatment of cardiovascular disorders or in cardioprotection after myocardial infarction; can be used as an immunomodulatory agent, especially in the treatment of autoimmune diseases such as arthritis, with For skin transplantation, organ transplantation, or similar surgical needs, for collagen disease, for allergy, or as an antineoplastic or antiviral agent; can be used to treat multiple sclerosis, Parkinson's disease (Parkinson's disease) and Huntington's chorea; may be used to treat depression, anxiety, stress-related disorders (eg, post-traumatic stress disorder, panic disorder, social phobia, or obsessive-compulsive disorder), premature ejaculation, psychosis, traumatic brain injury , stroke, Alzheimer's disease (Alzheimer's disease), spinal cord injury, drug addiction (eg, to treat alcohol, nicotine, opioids, or other drugs of abuse), or sympathetic nervous system disorders (eg, high blood pressure); may be used with Used to treat diarrhea; can be used to treat pulmonary conditions such as asthma, cough or pulmonary edema.

The pharmaceutically acceptable salts of the compounds described in the present disclosure are selected from inorganic salts or organic salts, and the compounds described in the present disclosure can react with acidic or basic substances to form corresponding salts.

On the other hand, the compounds of the present disclosure may exist in specific geometric or stereoisomeric forms. This disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to within the scope of this disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All such isomers, as well as mixtures thereof, are included within the scope of this disclosure.

In addition, the compounds and intermediates of the present disclosure may also exist in different tautomeric forms, and all such forms are included within the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (also known as proton tautomers) include interconversion via proton transfer, such as keto-enol and imine-enamine, lactam-lactam isomerizations . An example of a lactam-lactam equilibrium is between A and B as shown below.

All compounds in the present invention can be drawn as A or B type. All tautomeric forms are within the scope of the present invention. The naming of compounds does not exclude any tautomers.

The compounds of the present disclosure may be asymmetric, eg, have one or more stereoisomers. Unless otherwise specified, all stereoisomers include, such as enantiomers and diastereomers. Compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure or racemic forms. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or chiral reagents.

Optically active (R)- and (S)-isomers as well as D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present disclosure is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to provide pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, followed by conventional methods known in the art The diastereoisomers were resolved and the pure enantiomers recovered. In addition, separation of enantiomers and diastereomers is usually accomplished by the use of chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (eg, from amines to amino groups) formate).

The present disclosure also includes certain isotopically-labeled compounds of the present disclosure which are identical to those described herein, but wherein one or more atoms are replaced by an atom having an atomic weight or mass number different from that normally found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2H, ^3H , ^11C , ^13C , ^14C , ¹³ , ^respectively N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ¹²³ I, ¹²⁵ I and ³⁶ Cl and the like.

Unless otherwise stated, when a position is specifically designated as deuterium (D), the position is understood to have an abundance of deuterium (ie, at least 1000 times greater than the natural abundance of deuterium (which is 0.015%)) % of deuterium incorporated). Exemplary compounds having a natural abundance greater than deuterium may be at least 1000 times more abundant deuterium, at least 2000 times more abundant, at least 3000 times more abundant, at least 4000 times more abundant, at least 4000 times more abundant 5000 times more abundant deuterium, at least 6000 times more abundant deuterium or more abundant deuterium. The present disclosure also includes compounds of formula (I) in various deuterated forms. Each available hydrogen atom attached to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can refer to relevant literature to synthesize the compound of formula (I) in deuterated form. Commercially available deuterated starting materials can be used in preparing deuterated forms of compounds of formula (I), or they can be synthesized using conventional techniques using deuterated reagents including, but not limited to, deuterated borane, trideuterated Borane tetrahydrofuran solution, deuterated lithium aluminum hydride, deuterated iodoethane and deuterated iodomethane, etc.

"Optionally" or "optionally" means that the subsequently described event or circumstance can, but need not, occur, and that the description includes instances where the event or circumstance occurs or instances where it does not. For example, "C _1-6 alkyl optionally substituted by halogen or cyano" means that halogen or cyano may but need not be present, and the description includes the case where the alkyl is substituted by halogen or cyano and the case where the alkyl is not substituted by halogen and cyano substitution.

In the chemical structure of the compound of the present invention, the bond

Indicates an unspecified configuration, i.e. if a chiral isomer exists in the chemical structure, the bond

can be

or both

Two configurations. Although all of the above structural formulae are drawn as certain isomeric forms for simplicity, the present invention encompasses all isomers such as tautomers, rotamers, geometric isomers, diastereomers isomers, racemates and enantiomers. Terminology Explanation:

"Aromaticity" means that the ring structure is in a single plane, and the number of π electrons in the ring structure is [4n+2] according to Huckel's rule, where n represents an integer.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein, or a physiologically pharmaceutically acceptable salt or prodrug thereof, with other chemical components, and other components such as physiologically pharmaceutically acceptable carriers and excipients Form. The purpose of the pharmaceutical composition is to facilitate the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity.

"Pharmaceutically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, glidant, sweetener, diluent, preservative that has been approved by the US Food and Drug Administration as acceptable for use in humans or livestock animals , dyes/colorants, flavoring agents, surfactants, wetting agents, dispersing agents, suspending agents, stabilizers, isotonic agents, solvents or emulsifiers.

"Alkyl" refers to saturated aliphatic hydrocarbon groups, including straight and branched chain groups of 1 to 20 carbon atoms. An alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl and various branched chain isomers, etc. Unless otherwise specified, the alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from halogen, Deuterium, hydroxyl, oxo, nitro, cyano, C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, C _3-6 cycloalkane base, 3- to 6-membered heterocycloalkyl, C _5-8 cycloalkenyl, C _3-6 cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C _5-8 cycloalkenyloxy, C _{6- 10} -aryl or 5- to 6-membered heteroaryl, the C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, C _3-6 ring Alkyl, 3- to 6-membered heterocycloalkyl, C _5-8 cycloalkenyl, C _3-6 cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C _5-8 cycloalkenyloxy, C _{6 -10} aryl or 5 to 6 membered heteroaryl optionally substituted with one or more selected from halogen, deuterium, hydroxy, oxo, nitro, cyano.

"Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing 3 to 20 carbon atoms, preferably 3 to 8 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like; polycyclic cycloalkyl groups include spiro Ring, fused and bridged cycloalkyl groups.

The cycloalkyl ring can be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring linked to the parent structure is a cycloalkyl, non-limiting examples include indanyl, tetrahydronaphthalene base, benzocycloheptyl, etc. Unless otherwise specified, cycloalkyl can be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups, which are independently selected from deuterium, alkyl, alkenyl, alkyne group, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkane Oxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

"Monovalent group" refers to the "formal" elimination of a monovalent atom or group from a compound. "Subunit" refers to an atom or group of atoms formed by the "formal" elimination of two monovalent or one divalent compound.

"Alkylene ( _-CH2- )" means the remainder of the alkane molecule after removal of 2 hydrogen atoms, including straight and branched chain subgroups of 1 to 20 carbon atoms. An alkylene group containing 1 to 6 carbon atoms, non-limiting examples including methylene ( _-CH2- ), ethylene (eg _-CH2CH2- or -CH( _CH3 ₎ -), methylene Propyl (eg _-CH2CH2CH2- or -CH ₍ _CH2CH3 ₎ _- ₎ , butylene ₍ eg _{-CH2CH2CH2CH2-} ₎ . Unless otherwise specified, an alkylene group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from deuterium , aryl, heteroaryl, halogen substituted.

"Cycloalkylene" means the remainder of a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon containing 3 to 20 carbon atoms, preferably 3 to 8 carbon atoms, remaining after removal of 2 hydrogen atoms from a single carbon atom atom. Non-limiting examples of monocyclic cycloalkylene include cyclopropylene

Cyclobutylene

Cyclopentylene

cyclopentenylene

Cyclohexylene

cyclohexenylene

Unless otherwise specified, cycloalkylene may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups, which are independently selected from deuterium, alkyl, alkenyl, Alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycle Alkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

"Heterocyclyl" or "heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group containing from 3 to 20 ring atoms, one or more of which are selected from nitrogen, oxygen, or S (O) heteroatoms of _m (where m is an integer from 0 to 2), excluding ring moieties of -OO-, -OS- or -SS-, the remaining ring atoms being carbon. Preferably it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably 3 to 7 ring atoms. Non-limiting examples of monocyclic heterocycloalkyl include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piper pyridyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocycloalkyl groups include spiro, fused and bridged ring heterocycloalkyl groups. Non-limiting examples of "heterocycloalkyl" include:

and many more.

Heterocycloalkyl can be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from groups such as halogen, deuterium, hydroxy, oxo, nitro, cyano base, C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C _5-8 cycloalkenyl, C _3-6 cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C _5-8 cycloalkenyloxy, C _6-10 aryl or 5- to 6-membered heteroaryl , the C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl , C _5-8 cycloalkenyl, C _3-6 cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C _5-8 cycloalkenyloxy, C _6-10 aryl or 5- to 6-membered heteroaryl The group is optionally substituted with one or more groups selected from halogen, deuterium, hydroxy, oxo, nitro, cyano.

The heterocyclyl ring can be fused to an aromatic ring, a heteroaromatic ring, or a cyclic hydrocarbon, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include:

Wait.

"Aryl" or "aromatic ring" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (ie, rings that share adjacent pairs of carbon atoms) groups having a conjugated pi-electron system, preferably 6 to 12 elements such as phenyl and naphthyl.

Aryl may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from halogen, deuterium, hydroxyl, oxo, nitro, cyano, _{C1 -6} alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C _5-8 cycloalkenyl, C _3-6 cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C _5-8 cycloalkenyloxy, C _6-10 aryl or 5- to 6-membered heteroaryl, the C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C _{5- 8} -cycloalkenyl, C _3-6 cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C _5-8 cycloalkenyloxy, C _6-10 aryl or 5- to 6-membered heteroaryl are optionally One or more substituted by halogen, deuterium, hydroxyl, oxo, nitro, cyano.

The aryl ring may be fused to a heteroaromatic ring, a heterocyclic ring or a cyclic hydrocarbon, wherein the ring linked to the parent structure is an aryl ring, non-limiting examples of which include:

"Heteroaryl" or "heteroaromatic ring" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 6- to 12-membered, more preferably 5- or 6-membered. E.g. Non-limiting examples thereof include: imidazolyl, furanyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazine,

and many more.

Examples of nitrogen-containing heteroaryl groups include, but are not limited to, pyrrolyl, piperazinyl, pyrimidinyl, imidazolyl, pyridazinyl, pyrazinyl, tetrazolyl, triazolyl, pyridyl, pyrazolyl, oxa azolyl or thiazolyl, etc.

Heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from halogen, deuterium, hydroxy, oxo, nitro, cyano , C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C _5-8 cycloalkenyl, C _3-6 cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C _5-8 cycloalkenyloxy, C _6-10 aryl or 5- to 6-membered heteroaryl, The C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyloxy, C _2-6 alkynyloxy, C _3-6 cycloalkyl, 3- to 6-membered heterocycloalkyl, C _5-8 cycloalkenyl, C _3-6 cycloalkoxy, 3- to 6-membered heterocycloalkoxy, C _5-8 cycloalkenyloxy, C _6-10 aryl or 5- to 6-membered heteroaryl Optionally substituted with one or more selected from halogen, deuterium, hydroxy, oxo, nitro, cyano.

The heteroaryl ring can be fused to an aromatic ring, a heterocycle or a cyclic hydrocarbon, wherein the ring linked to the parent structure is a heteroaryl ring, non-limiting examples of which include:

"Alkoxy" refers to -O-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy. Similarly, "cycloalkoxy" is as defined above for "alkoxy".

"Halogen" refers to fluorine, chlorine, bromine or iodine.

"Substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently of one another, are substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the person skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.

"Substituted with one or more A, B..." means that it may be substituted with single or multiple substituents. When substituted by a plurality of substituents, it may be a plurality of the same substituents, or a combination of one or a plurality of different substituents.

Detailed ways

The present disclosure is further described below in conjunction with examples, but these examples do not limit the scope of the present disclosure.

The experimental methods without specific conditions in the examples of the present disclosure generally follow conventional conditions or conditions suggested by raw material or commodity manufacturers. Reagents with no specific source indicated are conventional reagents purchased in the market.

The structures of the compounds were determined by nuclear magnetic resonance (NMR) or/and mass spectrometry (MS). NMR shifts ([delta]) are given in units of 10<" ⁶ > (ppm). NMR was measured by Bruker AVANCE-400 nuclear magnetic instrument, and the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), and the internal standard was four Methylsilane (TMS). The spatial configuration of the optical isomers (isomers) of the compounds can be further confirmed by means of measuring single crystal parameters.

HPLC was measured using Waters ACQUITY ultra high performance LC, Shimadzu LC-20A systems, Shimadzu LC-2010HT series or Agilent Agilent 1200 LC high pressure liquid chromatograph (ACQUITY UPLC BEH C18 1.7UM 2.1X50MM column, Ultimate XB-C18 3.0* 150mm chromatographic column or Xtimate C18 2.1*30mm chromatographic column).

MS was measured with a Waters SQD2 mass spectrometer, scanned in positive/negative ion mode, and the mass scanning range was 100-1200.

Chiral HPLC analysis and determination using Chiralpak IC-3 100×4.6mm I.D., 3um, Chiralpak AD-3 150×4.6mm I.D., 3um, Chiralpak AD-3 50×4.6mm I.D., 3um, Chiralpak AS-3 150×4.6mm I.D., 3um, Chiralpak AS-3 100×4.6mm I.D., 3μm, ChiralCel OD-3 150×4.6mm I.D., 3um, Chiralcel OD-3 100×4.6mm I.D., 3μm, ChiralCel OJ-H 150×4.6mm I.D., 5um, Chiralcel OJ-3 150×4.6mm I.D., 3um chromatographic column;

The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, the size of the silica gel plate used for thin layer chromatography (TLC) is 0.15mm ~ 0.2mm, and the size of the TLC separation and purification products is 0.4mm ~0.5mm. The flash column purification system used Combiflash Rf150 (TELEDYNE ISCO) or Isolara one (Biotage). Forward column chromatography generally uses Yantai Huanghai silica gel 100-200 mesh, 200-300 mesh or 300-400 mesh silica gel as the carrier, or use Changzhou Santai pre-packed pre-packed ultra-pure normal phase silica gel column (40-63 μm, 60, 12 g ,, 25g, 40g, 80g or other specifications).

Reversed-phase column chromatography generally uses Changzhou Santai pre-packed ultra-pure C18 silica gel column (20-45μm,

40g, 80g, 120g, 220g or other specifications).

The high-pressure column purification system uses Waters AutoP, in conjunction with Waters XBridge BEH C18 OBD Prep Column,

5μm, 19mm X 150mm or Atlantis T3 OBD Prep Column,

5μm, 19mm X 150mm.

The chiral preparative column uses DAICEL CHIRALPAK IC (250mm*30mm, 10um) or Phenomenex-Amylose-1 (250mm*30mm, 5um).

The known starting materials in the present disclosure can be synthesized by using or according to methods known in the art, or can be purchased from Shanghai Titan Technology, ABCR GmbH & Co.KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology (Accela ChemBio Inc), Darui Chemicals and other companies.

There is no special description in the examples, and the reactions can all be carried out in an argon atmosphere or a nitrogen atmosphere.

Argon or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1 L.

Hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1 L.

The pressure hydrogenation reaction uses Parr 3916EKX hydrogenation apparatus and Qinglan QL-500 hydrogen generator or HC2-SS hydrogenation apparatus.

The hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times.

The microwave reaction used a CEM Discover-S 908860 microwave reactor.

There is no special description in the examples, and the solution refers to an aqueous solution.

There is no special description in the examples, and the reaction temperature is room temperature, which is 20°C to 30°C.

The monitoring of the reaction progress in the examples adopts thin layer chromatography (TLC), the developing solvent used in the reaction, the eluent system of column chromatography and the developing solvent system of thin layer chromatography used for purifying the compound, and the volume of the solvent The ratio is adjusted according to the polarity of the compound, and it can also be adjusted by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

Example 1

(S)-1-(2-((2-(6,8-Difluoro-4H-benzo[b][1,2,4]triazolo[4,3-d][1,4] Oxazin-7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholino)ethane-1-one (1)

Step 1: tert-butyl (2R)-2-[(methylsulfonyloxy)methyl]morpholine-4-carboxylate (1a)

To a solution of (2R)-tert-butyl 2-(hydroxymethyl)morpholine-4-carboxylate (100.00 g, 460.28 mmol) in toluene (1000 mL) was slowly added triethylamine at 0 °C under _N2 protection (96.0 mL, 690.42 mmol) and methanesulfonyl chloride (49.6 mL, 635.71 mmol) and stirred at room temperature. After the reaction was completed, the mixture was washed with water and saturated NaCl solution successively, the organic phase was dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure to obtain compound 1a (135.00 g, yield: 99.3%).

¹ H NMR (400 MHz, CDCl ₃ ): δ 4.24 (d, J=4.8 Hz, 2H), 3.92 (br d, J=11.0 Hz, 3H), 3.74-3.65 (m, 1H), 3.55 (dt, J=2.6, 11.6Hz, 1H), 3.08-3.06(m, 3H), 3.04-2.91(m, 1H), 2.79(br d, J=14.6Hz, 1H), 1.47(s, 9H).

Step 2: tert-butyl (2S)-2-[(1,3-dicarbonyl-2,3-dihydro-1H-isoindol-2-yl)methyl]morpholine-4-carboxylate ( 1b)

To a solution of compound 1a (135.00 g, 457.07 mmol) in N,N-dimethylacetamide (1350 mL) were sequentially added potassium phthalimide (93.12 g, 502.78 mmol) and tetrabutylammonium bromide ( 14.73 g, 45.71 mmol), stirred at 80°C. After the reaction was completed, the mixture was filtered, the filtrate was poured into water, and filtered to obtain a crude product. The crude product was stirred with NMP- _H2O (1/5, 2400 mL) at 25°C overnight, then filtered, and the filter cake was dried under reduced pressure to give compound 1b (120.00 g, 73.0%).

MS(ESI) m/z=369.1[M+Na] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.83-7.76 (m, 2H), 7.69-7.63 (m, 2H), 4.91 (spt, J=6.2 Hz, 3H), 3.96-3.84 (m, 1H) ,3.83-3.77(m,2H),3.75-3.64(m,2H),3.60(dd,J=4.6,13.6Hz,1H),3.37(dt,J=2.9,11.4Hz,1H),2.92(br s,1H),2.69(br s,1H),1.39(s,9H),1.21(s,11H).

Step 3: tert-Butyl (2S)-2-(aminomethyl)morpholine-4-carboxylate (1c)

To a solution of compound 1b (100.00 g, 288.70 mmol) in 2-methyltetrahydrofuran (1000 mL) was added hydrazine hydrate (24.8 mL, 433.05 mmol, 85% wt) at room temperature, followed by stirring at 80°C. After the reaction was completed, the mixture was cooled to room temperature, filtered, and the filtrate was washed with water and dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to obtain compound 1c (68.00 g, yield: 87.1%, purity: 80%).

¹ H NMR (400MHz, CDCl ₃ ): δ 6.27 (br s, 2H), 3.82 (br d, J=10.9Hz, 3H), 3.51-3.41 (m, 1H), 3.34-3.23 (m, 1H) ,3.29(br dd,J=3.7,6.7Hz,1H),2.85(br s,1H),2.73-2.63(m,2H),2.57(br s,1H),1.40(s,9H).

Step 4: tert-butyl (2S)-2-{[(4-methyl-2-nitrophenyl)amino]methyl}morpholine-4-carboxylate (1d)

To a solution of compound 1c (5.30 g, 24.50 mmol) in dimethyl sulfoxide (50 mL) was added 1-fluoro-4-methyl-2-nitrobenzene (3.80 g, 24.50 mmol) and calcium carbonate ( 2.94 g, 29.41 mmol), then stirred at 110 °C. After the reaction was completed, water was added to the reaction solution, extracted with ethyl acetate, the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product which was purified by flash silica gel chromatography to obtain compound 1d (4.00 g, yield rate: 46.4%).

MS(ESI) m/z=374.2[M+Na] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.10 (br s, 1H), 7.97 (d, J=1.0 Hz, 1H), 7.31-7.23 (m, 1H), 6.77 (d, J=8.6 Hz, 1H) ),4.04-3.76(m,3H),3.69(dq,J=3.9,6.8Hz,1H),3.57(dt,J=2.2,11.5Hz,1H),3.45-3.38(m,1H),3.37- 3.29(m, 1H), 3.37-3.29(m, 1H), 2.98(br s, 1H), 2.79(br s, 1H), 2.27(s, 3H), 1.47(s, 9H).

Step Five: 6,8-Difluoro-2H-benzo[b][1,4]oxazin-3(4H)-one (1e)

To a solution of 2-amino-4,6-difluorophenol (828.6 mg, 5.71 mmol) and potassium carbonate (2.4 g, 17.13 mmol) in tetrahydrofuran (20 mL) was added chloroacetyl chloride (0.5 mL, 6.28 mmol), followed by The reaction was stirred at 40°C. After the reaction was completed, it was extracted with dichloromethane and water, the organic phase was dried and concentrated, and the crude product was purified by silica gel column chromatography to obtain compound 1e (919.7 mg, yield: 87.0%).

MS(ESI) m/z=186.3[M+H] ⁺ .

Step Six: 6,8-Difluoro-3-carbonyl-3,4-dihydro-2H-benzo[b][1,4]oxazine-7-carbaldehyde (1f)

To a solution of compound le (3.70 g, 20.00 mmol) in tetrahydrofuran (10 mL) was added NaH (960.0 mg, 24.00 mmol, 60% wt) at 0 °C. After the mixture was stirred at 0 °C for 1 hour, n-butyllithium in n-hexane (12.0 mL, 30.00 mmol, 2.5 M) was added to the reaction solution at -78 °C. The reaction mixture was stirred at -78°C for 1 hour, then N,N-dimethylformamide (1.8 mL, 24.00 mmol) was added at -78°C and the reaction was kept stirring at -78°C. After the reaction was complete, the mixture was quenched with saturated NH ₄ Cl, extracted with ethyl acetate, the organic phase was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure, the crude product was purified by reverse phase column chromatography to give compound 1f (2.40 g, yield: 56.4%).

MS(ESI)m/z=214.3[M+H] ⁺ .

Step 7: tert-butyl (S)-2-((2-(6,8-difluoro-3-carbonyl-3,4-dihydro-2H-benzo[b][1,4]oxazine- 7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (1 g)

To a mixed solution of compound 1f (175.7 mg, 0.50 mmol) and compound 1d (106.6 mg, 0.5 mmol) in ethanol (6 mL) and water (1.2 mL) was added sodium hydrosulfite (261.2 mg, 1.50 mmol). The resulting mixture was stirred at 80°C under nitrogen protection. After the reaction was complete, the mixture was filtered, water was added to the filtrate, extracted with ethyl acetate, the combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to obtain compound 1 g (206.0 mg, yield: 80.0%).

MS(ESI) m/z=515.5[M+H] ⁺ .

¹ H NMR (400MHz, CDCl ₃ ): δ 9.33 (br s, 1H), 7.65 (s, 1H), 7.40 (d, J=8.4Hz, 1H), 7.19 (d, J=8.4Hz, 1H) ,6.53(br d,J＝7.2Hz,1H),4.71(s,2H),4.16-4.12(m,2H),4.11-4.05(m,1H),3.80(br s,1H),3.76-3.56 (m, 3H), 3.32(br s, 1H), 2.79(br s, 1H), 2.51(s, 3H), 1.41(s, 9H).

Step 8: tert-butyl (S)-2-((2-(6,8-difluoro-3-thio-3,4-dihydro-2H-benzo[b][1,4]oxazine -7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (1h)

To a solution of compound 1 g (411.6 mg, 0.80 mmol) in tetrahydrofuran (10 mL) was added Lawson's reagent (388.3 mg, 0.96 mmol) at room temperature, and the reaction mixture was stirred at room temperature. After the reaction was completed, the reaction solution was concentrated and purified by reverse-phase column chromatography to obtain compound 1h (371.4 mg, yield: 87.5%).

MS(ESI) m/z=531.5[M+H] ⁺ .

Step 9: tert-butyl(S,E)-2-((2-(6,8-difluoro-3-hydrazineidene-3,4-dihydro-2H-benzo[b][1,4 ]oxazin-7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (1i)

At room temperature, hydrazine hydrate (60 μL, 1.05 mmol, 85% wt) was added to a solution of compound 1h (371.4 mg, 0.70 mmol) in ethanol (4 mL), and stirred at 80°C. After the reaction was completed, the reaction solution was concentrated to obtain compound 1i (370.0 mg, yield: 100%).

Step ten: tert-butyl (S)-2-((2-(6,8-difluoro-4H-benzo[b][1,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholine-4-carboxylate (1j)

Compound 1i (370.0 mg, 0.70 mmol) was dissolved in triethyl orthoformate (7.5 mL) at room temperature, and the reaction mixture was stirred at 110 °C. After the reaction was completed, the reaction solution was concentrated and purified by reverse-phase column chromatography to obtain compound 1j (313.0 mg, yield: 83.0%).

MS(ESI) m/z=539.5[M+H] ⁺ .

Step eleven: (S)-1-(2-((2-(6,8-difluoro-4H-benzo[b][1,2,4]triazolo[4,3-d][ 1,4]oxazin-7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholino)ethan-1-one (1)

At room temperature, compound 1j (323.1 mg, 0.60 mmol) was dissolved in 1,4-dioxane (3 mL), p-toluenesulfonic acid monohydrate (1.14 g, 6.00 mmol) was added, and the mixture was heated at 100 °C under stirring. After the reaction was completed, methanol was added, and the pH was adjusted to about 8 with triethylamine at 0°C, then acetic anhydride (282 μL, 3.00 mmol) was added, and the mixture was stirred at room temperature. After the reaction was completed, the mixture was concentrated and the crude product was purified by reverse-phase column chromatography to obtain Compound 1 (64.0 mg, yield: 22.2%).

MS(ESI) m/z=481.2[M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ , t=75°C): δ 9.38 (s, 1H), 7.95 (dd, J=2.0, 9.5Hz, 1H), 7.65 (br d, J=8.3Hz, 1H), 7.52(s, 1H), 7.20(d, J=8.5Hz, 1H), 5.67(s, 2H), 4.33(dd, J=3.4, 15.2Hz, 1H), 4.16(br dd, J= 7.4, 15.2Hz, 1H), 3.73-3.39(m, 3H), 3.20(br s, 3H), 3.06-3.03(m, 1H), 2.47(s, 3H), 1.94(s, 3H).

Example 2

7-(1-(((S)-4-Acetylmorpholin-2-yl)methyl)-5-methyl-1H-benzo[d]imidazol-2-yl)-6,8-di Fluoro-1,2,9,9a-tetrahydro-3H-pyrrolo[1,2-a]indol-3-one (2)

Step 1: (R)-2-((4-acetylmorpholin-2-yl)methyl)isoindoline-1,3-dione (2a)

Compound 1b (8.00 g, 23.10 mmol) was dissolved in 1,4-dioxane (40 mL), 4N hydrochloric acid dioxane solution (46 mL, 184.77 mmol) was added in an ice-water bath, followed by stirring at room temperature. After the reaction was completed, the mixture was concentrated under reduced pressure, the obtained residue was dissolved in methanol (70 mL), triethylamine (13.5 mL, 97.45 mmol) and acetic anhydride (4.6 mL, 48.73 mmol) were slowly added sequentially under an ice-water bath, and the mixture was heated to room temperature. reaction. After the completion of the reaction, the reaction mixture was concentrated, ethyl acetate and water were added, and the layers were separated. The aqueous phase was washed with ethyl acetate, the combined organic phases were dried over anhydrous Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure to obtain the compound 2a (6.00 g, yield: 94.7%).

¹ H NMR (400MHz, CDCl ₃ ): δ 7.91-7.83 (m, 2H), 7.78-7.70 (m, 2H), 4.47 (br d, J=13.3Hz, 1H), 4.32 (br d, J= 13.6Hz, 1H), 3.96-3.86(m, 2H), 3.82-3.66(m, 3H), 3.60-3.39(m, 2H), 3.36-3.26(m, 1H), 3.13-3.01(m, 1H) ,2.89-2.79(m,1H),2.67(dd,J＝10.0,13.3Hz,1H),2.09(s,3H).

Step 2: (S)-1-(2-(aminomethyl)morpholino)ethane-1-one (2b)

To a solution of compound 2a (7.00 g, 24.28 mmol) in 2-methyltetrahydrofuran (70 mL) was added hydrazine hydrate (2.1 mL, 36.42 mmol, 85% wt) at room temperature, and stirred at 80°C. After the reaction was completed, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in ethyl acetate and filtered, and the filtrate was concentrated under reduced pressure to obtain compound 2b (3.00 g, yield: 66.4%).

¹ H NMR (400 MHz, CDCl ₃ ): δ 4.47-4.34 (m, 1H), 3.99-3.90 (m, 1H), 3.66-3.55 (m, 1H), 3.55-3.45 (m, 1H), 3.41- 3.19(m, 2H), 3.04-2.92(m, 1H), 2.83-2.75(m, 2H), 2.09(s, 3H).

Step 3: 1-[(2S)-2-{[(4-methyl-2-nitrophenyl)amino]methyl}morpholin-4-yl]ethane-1-one (2c)

To a solution of compound 2b (3.60 g, 13.65 mmol) in dimethyl sulfoxide (20 mL) was added 1-fluoro-4-methyl-2-nitrobenzene (2.12 g, 13.65 mmol) and calcium carbonate ( 1.64 g, 16.38 mmol), then the reaction mixture was stirred at 110 °C. After the reaction was completed, water was added, extracted with ethyl acetate, the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product which was purified by flash silica gel chromatography to obtain compound 2c (1.85 g, yield: 41.6%). ).

MS(ESI) m/z=294.1[M+H] ⁺ .

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.18-8.06 (m, 1H), 8.04-7.98 (m, 1H), 7.34-7.29 (m, 1H), 6.83-6.75 (m, 1H), 4.59- 4.39(m,1H),4.03(dd,J=2.6,10.7Hz,1H),3.79-3.55(m,3H),3.51-3.11(m,3H),2.91-2.63(m,1H),2.33- 2.27(m,3H),2.16-2.10(m,3H).

Step 4: 4-Bromo-3,5-difluoro-2-iodoaniline (2d)

4-Bromo-3,5-difluoroaniline (3.00 g, 14.42 mmol) was dissolved in acetic acid (30 mL), N-iodosuccinimide (4.22 g, 18.75 mmol) was added at room temperature, and the reaction mixture was Stir at 25°C. After the reaction was completed, the reaction mixture was poured into water, the mixture was extracted with ethyl acetate, and the combined organic layers were washed with 1N aqueous NaOH solution, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product by flash silica gel chromatography Purification gave compound 2d (1.60 g, yield: 29.9%).

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 6.58 (dd, J=1.8, 11.3 Hz, 1H), 6.03 (s, 2H).

Step 5: 4-Bromo-3,5-difluoro-2-(allyl)aniline (2e)

To a solution of compound 2d (900.0 mg, 2.70 mmol) in DMF (10 mL) was added allyltributyltin (0.67 mL, 2.156 mmol) followed by Pd(PPh ₃ ) ₄ (311.5 mg, 0.27 mmol), and the reaction mixture was purged with nitrogen After 3 replacements, the reaction was stirred at 100°C. After the reaction was complete, the mixture was quenched with saturated KF and filtered, the filtrate was extracted with ethyl acetate, the organic phase was washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography to give compound 2e (580.0 mg, yield: 69.4%).

¹ H NMR (400MHz, DMSO-d ₆ ): δ 6.43 (dd, J=1.8, 11.4 Hz, 1H), 5.87-5.76 (m, 1H), 5.73 (s, 2H), 5.03-4.94 (m, 2H), 3.27-3.21(m, 2H).

Step Six: 2-Bromo-N-[4-Bromo-3,5-difluoro-2-(allyl)phenyl]acetamide (2f)

To a solution of compound 2e (700.0 mg, 2.82 mmol) in dichloromethane (7 mL) at 0 °C was added K ₂ CO ₃ (0.32 mL, 5.64 mmol) and 2-bromoacetyl bromide (0.30 mL, 3.39 mmol), The reaction mixture was stirred at 25°C. After the reaction was completed, the reaction was quenched with ice water, the reaction solution was extracted with dichloromethane, the organic phase was washed with brine, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by flash silica gel chromatography to give compound 2f (640.0 mg, yield: 55.3%).

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 9.96 (s, 1H), 7.51 (dd, J=2.0, 10.4 Hz, 1H), 5.83 (tdd, J=6.0, 10.4, 16.8 Hz, 1H) ,5.05-4.91(m,2H),4.12(s,2H),3.47-3.42(m,2H).

Step 7: 7-Bromo-6,8-difluoro-1,2,9,9a-tetrahydro-3H-pyrrolo[1,2-a]indol-3-one (2g)

To a solution of compound 2f (640.0 mg, 1.734 mmol) in acetonitrile (25 mL) at 0°C were sequentially added Pd(OAc) ₂ (38.9 mg, 0.17 mmol) and K ₂ CO ₃ (719.1 mg, 5.20 mmol). After the reaction mixture was purged with nitrogen three times, it was stirred at 40°C. After the reaction was completed, water was added to the reaction mixture, the reaction mixture was extracted with ethyl acetate, the organic phase was washed with brine, dried over anhydrous Na ₂ SO ₄ and filtered, and the filtrate was concentrated to obtain a residue. The residue was purified by flash silica gel chromatography to give compound 2g (380.0 mg, yield: 68.4%).

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.25 (dd, J=1.3, 8.0 Hz, 1H), 4.78-4.66 (m, 1H), 3.30 (dd, J=8.8, 15.8 Hz, 1H), 2.95 -2.78(m,2H),2.67-2.48(m,2H),2.11-1.97(m,1H).

Step 8: 6,8-Difluoro-3-carbonyl-2,3,9,9a-tetrahydro-1H-pyrrolo[1,2-a]indole-7-carbaldehyde (2h)

To a solution of compound 2g (200.0 mg, 0.69 mmol) in tetrahydrofuran (5 mL) at 0°C was added NaH (83.3 mg, 2.08 mmol), and after stirring at 0°C for 1 hour, n-butyl was added at -78°C. A solution of lithium in n-hexane (0.69 mL, 1.74 mmol, 2.5 M) was added to the reaction solution. The reaction mixture was stirred at -78°C for 1 hour, then N,N-dimethylformamide (0.27 mL, 3.47 mmol) was added at -78°C and the reaction mixture was stirred at -78°C. After the reaction was complete, the mixture was quenched with saturated NH ₄ Cl, extracted with ethyl acetate (, the organic phase was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give compound 2h (150.0 mg, yield: 91.6%).

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.09 (s, 1H), 7.13 (br d, J=10.5Hz, 1H), 4.76 (br dd, J=6.4, 9.2Hz, 1H), 2.98 -2.81(m,3H),2.20-2.01(m,4H).

Step nine: 7-(1-(((S)-4-acetylmorpholin-2-yl)methyl)-5-methyl-1H-benzo[d]imidazol-2-yl)-6, 8-Difluoro-1,2,9,9a-tetrahydro-3H-pyrrolo[1,2-a]indol-3-one (2)

To a mixed solution of compound 2h (200.0 mg, 0.84 mmol) and compound 2c (247.3 mg, 0.84 mmol) in ethanol (6 mL) and water (2 mL) was added sodium hydrosulfite (440.4 mg, 2.53 mmol). The resulting mixture was stirred under nitrogen protection at 80°C. After the reaction was complete, the mixture was filtered, water was added to the filtrate, extracted with ethyl acetate, the combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, and the obtained residue was purified by preparative HPLC to give compound 2 (50.0 mg, yield: 12.3%).

MS(ESI) m/z=481.2[M+H] ⁺ .

¹ H NMR (400MHz, DMSO-d ₆ ): δ 7.71-7.58 (m, 1H), 7.53-7.45 (m, 1H), 7.25-7.03 (m, 2H), 4.90-4.74 (m, 1H), 4.37-3.96(m, 3H), 3.85-3.72(m, 1H), 3.68-3.49(m, 2H), 3.32-3.19(m, 2H), 3.11(br t, J=11.7Hz, 1H), 3.03 -2.82(m,2H),2.80-2.62(m,1H),2.46-2.39(m,4H),2.31-2.04(m,2H),2.00-1.90(m,3H).

Example 3

(S)-1-(2-((2-(6,8-Difluoro-4H-benzo[b][1,2,4]triazolo[4,3-d][1,4] Oxazin-7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholino)ethan-1-one (3)

Step 1: 1,5-Difluoro-3-nitro-2-(allyloxy)benzene (3a)

To a solution of 2,4-difluoro-6-nitrophenol (3.00 g, 17.13 mmol) and potassium carbonate (4.74 g, 20.56 mmol) in acetonitrile (30 mL) was added allyl bromide (1.8 mL, 20.56 mL) at room temperature mmol). The reaction mixture was stirred at 60°C. After the reaction was complete, water was added to the mixture, extracted with ethyl acetate, and the combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to obtain a crude product which was purified by silica gel column chromatography to obtain compound 3a (4.00 g, yield: 97.6%).

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 7.89-7.77 (m, 2H), 6.06-5.92 (m, 1H), 5.42-5.24 (m, 2H), 4.66 (dd, J=0.9, 5.9 Hz, 2H).

Step 2: 1,5-Difluoro-2-(allyloxy)aniline (3b)

To a mixed solution of compound 3a (2.00 g, 9.30 mmol) in tetrahydrofuran (8 mL), ethanol (8 mL) and water (8 mL) was added NH ₄ Cl (4.93 g, 92.95 mmol) and iron powder (5.19 g, 92.95 mmol) at room temperature 92.95 mmol) and the resulting mixture was stirred at 70 °C. After the reaction was complete, the reaction mixture was filtered, the filtrate was extracted with ethyl acetate, the combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to obtain compound 3b (1.80 g, yield: 94.2%).

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 6.34-6.20 (m, 2H), 6.13-5.99 (m, 1H), 5.49 (br s, 2H), 5.31 (dd, J=1.5, 17.3 Hz ,1H),5.20(br d,J＝10.3Hz,1H),4.38(d,J＝5.8Hz,2H).

Step 3: 2-Bromo-N-[3,5-difluoro-2-(allyloxy)phenyl]acetamide (3c)

To a solution of compound 3b (1.20 g, 6.48 mmol) in dichloromethane (10 mL) at 0 °C was added K ₂ CO ₃ (1.79 g, 12.96 mmol) and 2-bromoacetyl bromide (0.68 mL, 7.78 mmol), Stir at room temperature. After the reaction was complete, water was added to quench the reaction and extracted with dichloromethane. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated, and the crude product was purified by silica gel column chromatography to obtain compound 3c (1.30 g, yield: 59.0%).

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 9.97 (s, 1H), 7.75 (td, J=2.5, 10.9 Hz, 1H), 7.09 (tdd, J=2.4, 8.9, 11.2 Hz, 1H) ,6.14-5.99(m,1H),5.37-5.17(m,2H),4.54(d,J=6.3Hz,2H),4.26-4.21(m,2H).

Step 4: 6,8-Difluoro-2,3,3a,4-tetrahydro-1H-benzo[b]pyrrolo[1,2-d][1,4]oxazin-1-one (3d )

To a solution of compound 3c (1.30 g, 4.25 mmol) in acetonitrile (30 mL) at 0°C were sequentially added Pd(OAc) ₂ (100.0 mg, 0.42 mmol) and K ₂ CO ₃ (1.76 g, 12.74 mmol). After the reaction mixture was purged with nitrogen three times, the reaction was stirred at 40°C. After the reaction was completed, water was added to the reaction mixture, followed by extraction with ethyl acetate. The organic phase was washed with brine, dried _over anhydrous _Na2SO4 and filtered, and the filtrate was concentrated to give a residue. The residue was purified by flash silica gel chromatography to give compound 3d (720.0 mg, yield: 67.8%).

¹ H NMR (400MHz, CDCl ₃ ): δ 8.22-8.10 (m, 1H), 6.64 (ddd, J=3.1, 8.2, 10.9Hz, 1H), 4.59 (dd, J=3.1, 10.8Hz, 1H) ,4.13-4.01(m,1H),3.80-3.68(m,1H),2.76-2.63(m,1H),2.60-2.49(m,1H),2.35(dddd,J=1.7,7.0,9.0,12.8 Hz,1H),1.73(tdd,J=9.6,11.6,12.6Hz,1H).

Step 5: 6,8-Difluoro-3-carbonyl-2,3,9,9a-tetrahydro-1H-pyrrolo[1,2-a]indole-7-carbaldehyde (3e)

To a solution of compound 3d (300.0 mg, 1.33 mmol) in tetrahydrofuran (4 mL) was slowly added a solution of lithium diisopropylamide in n-hexane (3.3 mL, 6.66 mmol, 2 M) at -78°C. After the reaction mixture was stirred at -78°C for 0.5 hours, N,N-dimethylformamide (0.52 mL, 6.66 mmol) was added at -78°C, and the reaction mixture was stirred at -78°C. After the reaction was complete, the mixture was quenched with saturated _NH4Cl and extracted with ethyl acetate. The organic phase was dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give compound 3e (300.0 mg, yield: 88.9%).

¹ H NMR (400MHz, DMSO-d ₆ ): δ 10.11 (s, 1H), 8.21-8.10 (m, 1H), 4.74-4.66 (m, 1H), 4.15-4.06 (m, 1H), 3.87- 3.77(m,1H),2.68(ddd,J=9.3,11.5,17.1Hz,1H),2.41(dd,J=8.4,17.2Hz,1H),2.28-2.19(m,1H),1.79-1.67( m,1H).

Step 6: (S)-1-(2-((2-(6,8-Difluoro-4H-benzo[b][1,2,4]triazolo[4,3-d][1 ,4]oxazin-7-yl)-5-methyl-1H-benzo[d]imidazol-1-yl)methyl)morpholino)ethan-1-one (3)

To a mixed solution of compound 3e (250.0 mg, 0.99 mmol) and compound 2c (289.6 mg, 0.99 mmol) in ethanol (3 mL) and water (1 mL) was added sodium hydrosulfite (515.7 mg, 2.96 mmol). The resulting mixture was stirred at 80°C under nitrogen protection. After the reaction was completed, the mixture was filtered, and water was added to the filtrate, followed by extraction with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated, and the obtained residue was purified by preparative HPLC to give compound 3 (200.0 mg, yield: 40.7%).

MS(ESI) m/z=497.2[M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.24 (dd, J=2.1, 11.6 Hz, 1H), 7.60 (br d, J=7.9 Hz, 1H), 7.49 (s, 1H), 7.19- 7.12(m, 1H), 4.70(dd, J=3.2, 10.7Hz, 1H), 4.32-4.07(m, 3H), 3.94-3.84(m, 1H), 3.70-3.45(m, 3H), 3.30- 3.15(m, 1H), 2.69(ddd, J=9.3, 11.1, 17.1Hz, 2H), 2.49-2.39(m, 4H), 2.36-2.24(m, 2H), 1.93(s, 3H), 1.85- 1.71(m,1H).

Compound 3 (150.0 mg) was resolved by SFC (column: DAICEL CHIRALCEL OJ 250 mm×30 mm×10 μm, 0.1% NH ₃ ^. H ₂ O in ethanol: 25%-25%) to obtain compounds 3-E1 (50.2 mg) and 3 -E2 (53.2 mg). Chiral analysis method: Column: Chiralcel OJ-3 100×4.6 mm ID, 3 μm; Mobile Phase: A: CO ₂ , B: Ethanol (0.05% DEA); Gradient: 5% to 40% of B in 4 min; Flow Rate: 2.8mL/min.

Compound 3-E1:

Chiral supercritical fluid chromatography (Chiral SFC): t _R =2.95min.

MS(ESI) m/z=497.2[M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ , t=75°C): δ 8.24 (dd, J=2.4, 11.6 Hz, 1H), 7.60 (br d, J=8.0 Hz, 1H), 7.49 (s, 1H), 7.16(d, J＝8.0Hz, 1H), 4.70(dd, J＝3.2, 10.4Hz, 1H), 4.32-4.05(m, 4H), 3.88(t, J＝10.3Hz, 1H), 3.68-3.44(m, 3H), 3.23(br s, 1H), 2.69(ddd, J=9.4, 11.0, 17.1Hz, 2H), 2.48-2.39(m, 4H), 2.38-2.20(m, 2H) ,1.93(s,3H),1.85-1.71(m,1H).

Compound 3-E2:

Chiral supercritical fluid chromatography (Chiral SFC): t _R =3.10min.

MS(ESI) m/z=497.2[M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ , t=75°C): δ 8.24 (dd, J=2.0, 11.6 Hz, 1H), 7.60 (br d, J=8.0 Hz, 1H), 7.49 (s, 1H), 7.16(d, J＝7.2Hz, 1H), 4.70(dd, J＝3.2, 10.8Hz, 1H), 4.33-4.04(m, 4H), 3.88(t, J＝10.3Hz, 1H), 3.70-3.45(m,3H),3.30-3.17(m,1H),2.69(ddd,J=9.4,11.1,17.1Hz,2H),2.48-2.39(m,4H),2.37-2.22(m,2H) ),1.93(s,3H),1.84-1.70(m,1H).

Example 4

Methyl(2S)-2-((2-(6,8-Difluoro-1-carbonyl-2,3,3a,4-tetrahydro-1H-benzo[b]pyrrolo[1,2-d ][1,4]oxazin-7-yl)-7-methylimidazo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylate (4)

In a microwave tube, (2S)-methyl 2-ethynylmorpholine-4-carboxylate (33.4 mg, 0.20 mmol), compound 3e (50.0 mg, 0.20 mmol) and 4-methylpyridin-2-amine were combined (21.4 mg, 0.20 mmol) was dissolved in toluene (2 mL), then N,N-dimethylacetamide (6 μL, 0.06 mmol), cuprous chloride (5.9 mg, 0.06 mmol) and copper triflate were added (21.4 mg, 0.06 mmol), the reaction solution was heated to 120°C under microwave (1 bar) to react. After the reaction was completed, water and ammonia water were added to the reaction mixture, and the reaction mixture was extracted with dichloromethane. The combined organic phases were washed with brine, dried over anhydrous Na ₂ SO ₄ and filtered, the filtrate was concentrated, and the resulting residue was purified by flash silica gel chromatography to give compound 4 (50.0 mg, yield: 49.4%). Compound 4 (50.0 mg) was resolved by SFC (column: DAICEL CHIRALCEL OJ 250 mm×30 mm×10 μm, 0.1% NH ₃ ^. H ₂ O in ethanol: 25%-25%) to obtain compounds 4-E1 (20.8 mg) and 4 -E2 (17.4 mg). Chiral analysis method: Column: Chiralcel OJ-3 100×4.6 mm ID, 3 μm; Mobile Phase: A: CO ₂ , B: Ethanol (0.05% DEA); Gradient: 5% to 40% of B in 4 min; Flow Rate: 2.8mL/min.

Compound 4-E1:

Chiral supercritical fluid chromatography (Chiral SFC): t _R =2.81min.

MS(ESI) m/z=513.3[M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.39 (d, J=7.0 Hz, 1H), 8.17 (dd, J=2.0, 11.6 Hz, 1H), 7.33 (s, 1H), 6.82 (dd , J=1.6, 7.2Hz, 1H), 4.74-4.59(m, 1H), 4.21-4.04(m, 1H), 3.87(t, J=10.3Hz, 1H), 3.79-3.63(m, 3H), 3.55(s, 3H), 3.50-3.42(m, 1H), 3.28-3.19(m, 1H), 3.08-2.94(m, 2H), 2.82(br s, 1H), 2.67(ddd, J=9.4, 11.1,17.0Hz,2H),2.45-2.34(m,4H),2.30-2.19(m,1H),1.84-1.65(m,1H).

Compound 4-E2:

Chiral supercritical fluid chromatography (Chiral SFC): t _R =2.91min.

MS(ESI) m/z=513.3[M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.39 (d, J=7.1 Hz, 1H), 8.17 (dd, J=2.0, 11.5 Hz, 1H), 7.33 (s, 1H), 6.81 (dd ,J=1.5,7.1Hz,1H),4.67(dd,J=3.2,10.8Hz,1H),4.19-4.07(m,1H),3.85(t,J=10.3Hz,1H),3.77-3.59( m, 3H), 3.55(s, 3H), 3.52-3.42(m, 1H), 3.29-3.19(m, 1H), 3.01(br d, J=6.2Hz, 2H), 2.81(br s, 1H) ,2.67(ddd,J=9.4,11.1,17.0Hz,2H),2.46-2.34(m,4H),2.30-2.18(m,1H),1.81-1.66(m,1H).

Example 5

7-(3-(((S)-4-Acetylmorpholin-2-yl)methyl)-7-methylimidazo[1,2-a]pyridin-2-yl)-6,8- Difluoro-2,3,3a,4-tetrahydro-1H-benzo[b]pyrrolo[1,2-d][1,4]oxazin-1-one (5)

In a microwave tube, combine (S)-1-(2-ethynylmorpholino)ethan-1-one (78.6 mg, 0.51 mmol), compound 3e (130.0 mg, 0.51 mmol) and 4-picoline -2-amine (55.5 mg, 0.0.51 mmol) was dissolved in toluene (3 mL), then N,N-dimethylacetamide (14 μL, 0.15 mmol), cuprous chloride (15.2 mg, 0.15 mmol) and trimethylacetamide (14 μL, 0.15 mmol) were added. Copper fluoromethanesulfonate (55.7 mg, 0.15 mmol). The resulting mixture was then sparged with nitrogen for 5 minutes and heated to 120°C under microwave (1 bar) for 5 hours. After the reaction was completed, water (5 mL) and ammonia water (10 mL) were added to the reaction mixture, and the reaction mixture was extracted with dichloromethane (20 mL×3). The combined organic phases were washed with brine, dried over anhydrous Na ₂ SO ₄ and filtered, the filtrate was concentrated, and the resulting residue was subjected to preparative HPLC (column: Boston Prime C18 150*30mm*5 μm, acetonitrile/water (with 0.05% ammonia) = 30-55%, 9 min) to give compound 5 (50.0 mg, yield: 13.4%) after purification and lyophilization. Compound 5 (50.0 mg) was subjected to the first SFC (column: DAICEL CHIRALCEL OD 250 mm x 30 mm x 10 μm, 0.1% NH ₃ ^. H ₂ O in ethanol: 40%-40%) and the second SFC (column: Phenomenex- Cellulose-2 250mm×30mm×10μm, 0.1% NH ₃ ^. H ₂ O in ethanol: 45%-45%) was resolved to obtain compounds 5-E1 (4.2 mg) and 5-E2 (8.6 mg).

Compound 5-E1: Chiral Analysis Method: Column: Cellulose-2 100×4.6 mm ID, 3 μm; Mobile Phase: A: CO ₂ , B: Ethanol (0.05% DEA); Gradient: 50% of B in 7 min; Flow Rate : 2.8mL/min.

Chiral supercritical fluid chromatography (Chiral SFC): t _R =4.13min.

MS(ESI) m/z=497.2[M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.38 (d, J=7.2 Hz, 1H), 8.16 (dd, J=2.0, 11.6 Hz, 1H), 7.33 (s, 1H), 6.82 (dd ,J=1.2,7.2Hz,1H),4.67(dd,J=3.2,10.8Hz,1H),4.17-4.01(m,2H),3.86(t,J=10.0Hz,1H),3.71(br dd , J=2.4, 11.6Hz, 1H), 3.65-3.37(m, 2H), 3.04(br dd, J=4.0, 6.0Hz, 2H), 2.73-2.52(m, 2H), 2.46-2.36(m, 4H), 2.35-2.22(m, 2H), 1.96-1.85(m, 3H), 1.81-1.71(m, 1H), 1.30-1.22(m, 1H).

Compound 5-E2: Chiral Analysis Method: Column: Chiralcel OD-3 150×4.6 mm ID, 3 μm; Mobile Phase: A: CO ₂ , B: Isopropanol (0.05% DEA); Gradient: 40% of B in 7min; flow rate: 2.5mL/min.

Chiral supercritical fluid chromatography (Chiral SFC): t _R =3.96min.

MS(ESI) m/z=497.2[M+H] ⁺ .

¹ H NMR (400 MHz, DMSO-d ₆ ): δ 8.36 (d, J=7.2 Hz, 1H), 8.16 (dd, J=2.0, 11.6 Hz, 1H), 7.31 (d, J=0.8 Hz, 1H) ),6.80(dd,J＝1.6,7.2Hz,1H),4.67(dd,J＝3.2,10.8Hz,1H),4.13(ddt,J＝3.2,6.8,9.6Hz,2H),3.88-3.82( m,1H),3.74-3.69(m,1H),3.60-3.43(m,2H),3.05-3.02(m,2H),2.74-2.61(m,2H),2.45-2.40(m,1H), 2.39(d, J＝0.8Hz, 3H), 2.35-2.24(m, 2H), 1.91(br s, 3H), 1.81-1.71(m, 1H), 1.28-1.26(m, 1H).

biological test

Test Case A. Evaluation of Biological Activity in Vitro

Compounds were screened for antagonistic activity at hP2X3 and hP2X2/3 receptors by FLIPR assay (effects of compounds on ion channels were expressed as changes in calcium flux signals).

1. Experimental equipment and materials

2. Experimental steps

Digest 1321N1 cells (adherent cells) stably transfected with hP2X3 and hP2X2/3 receptors, resuspend and count in plating medium (DMEM+10% DFBS) after centrifugation, and adjust cells to 2*10 ⁵ cells/mL , in 384-well Assay Plate, spread 50 μL of cells per well, and place in 5% CO ₂ , 37 ℃ incubator for 16-24 hours. Prepare 180 times the required concentration of the test compound (20mM DMSO stock solution) with DMSO, add 500nL per well to the 384-well Compound Plate, and supplement with 30μL FLIPR buffer (1*HBSS+2mM _CaCl2 containing 1.26mM Ca2 ⁺ ). +20 mM HEPES), shake for 20-40 min to mix. Prepare 3 times the required concentration of agonist α,β-meATP (500nM for hP2X3 cells and 1000nM for hP2X2/3 cells) with FLIPR buffer, and add 35μL of agonist to another 384-well Compound Plate per well. Take out the cell plate that has been cultured for 16-24 hours, aspirate the cell supernatant, and add 30 μL Dye (

Calcium 4 Assay Kit, diluted in FLIPR buffer), incubate for 1 hour. 15 μL of compound (loaded by FLIPR instrument) was added to each well of cells, and 15 minutes later, 22.5 μL of agonist was added to each well to detect fluorescence signals (excitation wavelength 470nm-495nm, emission wavelength 515nm-575nm). The difference between the signal peak and the trough was taken as the basic data, the highest concentration of the positive drug was taken as the 100% inhibition rate, and the DMSO data was taken as the 0% inhibition rate, the inhibitory effect curve of the compound was fitted on the software Graphpad Prism 6 and the IC ₅₀ was calculated value. The results are shown in Table 1.

Table 1. The median inhibitory concentration (IC ₅₀ ) of the compounds of the present invention on hP2X3 and hP2X2/3 receptors

化合物编号Compound number	hP2X3(IC ₅₀,nM) hP2X3( _IC50 ,nM)	hP2X2/3(IC ₅₀,nM) hP2X2/3( _IC50 ,nM)
MK-7264MK-7264	35.435.4	116.2116.2
11	123.6123.6	NTNT
22	103.8103.8	NTNT
33	47.347.3	NTNT
3-E13-E1	30.430.4	21462146
3-E23-E2	82.182.1	55985598
4-E14-E1	68.468.4	NTNT
4-E24-E2	131.0131.0	NTNT
5-E15-E1	62.762.7	1285012850
5-E25-E2	92.992.9	NTNT

NT: Not Tested.

Claims

A compound of formula I or a pharmaceutically acceptable salt thereof:

Wherein, ring A and ring B are independently selected from 6-membered aromatic ring, 5-7-membered heteroaromatic ring and 5-7-membered heterocyclic ring, and ring A and ring B are connected by sharing 2 atoms;

Y is
Wherein, ring C is a 5-8 membered heterocyclic ring or a 5-8 membered heteroaromatic ring, and ring D is a 5-8 membered heterocyclic ring or a 5-8 membered heteroaromatic ring, and ring C and ring D share a nitrogen atom and 1 carbon atom is connected, and ring D and the benzene ring are connected by sharing 2 carbon atoms;

Q is

R 3 is independently selected from hydrogen, deuterium, halogen, C 1-4 alkyl, C 3-8 cycloalkyl and C 3-8 cycloalkylene, the C 1-4 alkyl, C 3-8 ring Alkyl and C 3-8 cycloalkane are optionally substituted with one or more deuterium or halogen; alternatively, R on adjacent carbon atoms forms a C 3- optionally substituted with one or more deuterium or halogen 8 cycloalkanes;

R 4 is selected from hydrogen, deuterium and the following groups optionally substituted with one or more substituents: C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl and C 3-8 cycloalkylmethylene, the substituents being selected from deuterium, halogen, -OH and -CN;

X is selected from -O-, -CR a1 R a2 - and -NR b -;

R 5a and R 5b are independently selected from hydrogen, deuterium, halogen and C 1-6 alkyl optionally substituted with deuterium or halogen;

R a1 and R a2 are independently selected from hydrogen, deuterium, halogen, C 1-6 alkyl optionally substituted with one or more deuterium or halogen, and C 3-8 optionally substituted with one or more deuterium or halogen Cycloalkyl; or, R a1 , R a2 and the carbon atoms to which they are commonly attached form a C 3-6 cycloalkylene optionally substituted by one or more deuterium or halogen;

R b is selected from hydrogen, deuterium and the following groups optionally substituted with one or more substituents: C 1-6 alkyl, -C(=O)-C 1-4 alkyl and -S(=O) 2 -C 1-4 alkyl, the substituent is selected from: deuterium, halogen, -OH and -CN;

m is an integer selected from 0 to 9;

n is an integer selected from 0-7.
The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound shown in formula I is:

Wherein, Z 1 is selected from C, CR', O, N and NR", Z 2 , Z 3 , Z 4 , Z 5 , Z 6 and Z 7 are independently C or N, and ring A and ring B are aromatic ;

R 1 is selected from the group consisting of hydrogen, deuterium, halogen and the following groups optionally substituted with one or more substituents: C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl and C 3 -8 cycloalkylmethylene, the substituent is selected from: deuterium, halogen, -OH and -CN;

R 2 is independently selected from hydrogen, deuterium, halogen, C 1-6 alkyl and C 3-8 cycloalkyl, which are optionally replaced by one or more substituted by deuterium, halogen or -OH;

R' is selected from hydrogen, deuterium, halogen and C 1-4 alkyl optionally substituted with one or more deuterium or halogen;

R" is selected from hydrogen, deuterium, C 1 -C 6 alkyl and C 3-8 cycloalkyl, said C 1 -C 4 alkyl and C 3-8 cycloalkyl optionally being substituted by one or more deuterium or halogen substitution;

m is an integer selected from 0 to 3; Y and Q are as defined in claim 1 .
The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein Z 1 is N or O; Z 1 is preferably N.
According to the arbitrary described compound of claim 1-3 or its pharmaceutically acceptable salt, wherein compound shown in formula I is selected from:

R 1 , R 2 , m, Y, Q are as defined in claim 2 .
The compound according to claim 4 or a pharmaceutically acceptable salt thereof, wherein the compound represented by formula I is
The compound according to claim 4 or a pharmaceutically acceptable salt thereof, wherein the compound represented by formula I is
The compound according to any one of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein Ring C is
And ring C is a 5-8 membered heterocycle; preferably ring C is
The compound according to claim 7, or a pharmaceutically acceptable salt thereof, wherein Ring C is selected from

R 6 is independently selected from hydrogen, deuterium, halogen, C 1-6 alkyl and C 3-6 cycloalkylene, said C 1-6 alkyl and C 3-6 cycloalkylene are optionally replaced by one or Multiple deuterium or halogen substitution; alternatively, R on adjacent carbon atoms forms C cycloalkane optionally substituted with one or more deuterium or halogen;

p is an integer selected from 1-3, and q is an integer selected from 0-9.
The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein ring C is
preferred

most preferred
A compound according to claim 8 or 9, or a pharmaceutically acceptable salt thereof, wherein R 6 is independently selected from hydrogen, deuterium, halogen and C 1-4 alkyl optionally substituted with one or more deuterium or halogen; preferably R6 is independently selected from hydrogen, deuterium and halogen ; more preferably R6 is independently hydrogen or deuterium.
The compound according to any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein Ring C is
The compound according to any one of claims 1-6 or a pharmaceutically acceptable salt thereof, wherein ring C is a 5-membered heteroaromatic ring, preferably ring C is
c 1 , c 2 and c 3 are each independently CR' or N, said R' as defined in claim 2, more preferably
most preferred
most preferred
The compound according to any one of claims 1-12, or a pharmaceutically acceptable salt thereof, wherein Ring D is selected from

R 7 is independently selected from hydrogen, deuterium, halogen, C 1-6 alkyl, C 3-6 cycloalkyl and C 3-6 cycloalkylene, the C 1-6 alkyl, C 3-6 ring Alkyl and C3-6cycloalkylene are optionally substituted with one or more deuterium or halogen; alternatively, R7 on adjacent carbon atoms forms C3 optionally substituted with one or more deuterium or halogen -6 cycloalkanes,

r is an integer selected from 0-3; s is an integer selected from 0-7, further, ring D is preferably
The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, wherein Y is selected from:

preferred
A compound according to any one of claims 1-14, or a pharmaceutically acceptable salt thereof, wherein R5a and R5b are independently selected from hydrogen, deuterium and halogen.
The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R 5a and R 5b are independently selected from hydrogen, fluorine and chlorine; preferably both R 5a and R 5b are fluorine.
A compound according to any one of claims 2-16, or a pharmaceutically acceptable salt thereof, wherein R 1 is selected from the group consisting of hydrogen, deuterium, halogen and the following groups optionally substituted with one or more substituents: C 1-6 alkyl and C 3-8 cycloalkylmethylene, the substituent is deuterium or halogen, further, R 1 is preferably selected from hydrogen, deuterium, halogen and C 1- optionally substituted by one or more deuterium or halogen 6 alkyl; more preferably R 1 is methyl optionally substituted with one or more deuterium or halogen.
The compound according to any one of claims 2-17, or a pharmaceutically acceptable salt thereof, wherein R 2 is independently selected from hydrogen, deuterium, halogen, and C 1-6 alkyl optionally substituted by a or more deuterium or halogen substitution; preferably R2 is independently selected from hydrogen , deuterium and halogen.
The compound according to any one of claims 1-18, or a pharmaceutically acceptable salt thereof, wherein Q is
The compound according to any one of claims 1-19, or a pharmaceutically acceptable salt thereof, wherein R is independently selected from hydrogen, deuterium, halogen, C 1-4 alkyl and C 3-8 cycloalkylene, the C 1-4 alkyl and C 3-8 cycloalkylene are optionally substituted by one or more deuterium or halogen; or, R on adjacent carbon atoms is optionally substituted by one or more deuterium or halogen of C 3-8 cycloalkanes.
The compound of claim 20, or a pharmaceutically acceptable salt thereof, wherein R is independently selected from hydrogen, deuterium, halogen, and C 1-4 alkyl optionally substituted by one or more of Deuterium or halogen substitution; alternatively, R on adjacent carbon atoms forms a C cycloalkane optionally substituted with one or more deuterium or halogen;

Preferably R3 is independently selected from hydrogen, deuterium and halogen.
The compound according to any one of claims 1-21, or a pharmaceutically acceptable salt thereof, wherein R4 is selected from the group consisting of hydrogen, deuterium and the following groups optionally substituted with one or more substituents: C1-6 alkyl, C 1-6 alkoxy and C 3-8 cycloalkyl, the substituents are deuterium or halogen, further, R 4 is preferably the following group C 1-4 alkane optionally substituted by one or more substituents or C 1-4 alkoxy, the substituent is deuterium or halogen, R 4 is more preferably methyl, ethyl, or methoxy, and R 4 is most preferably methyl or methoxy.
The compound according to any one of claims 1-22, or a pharmaceutically acceptable salt thereof, wherein X is selected from -O-, -CR a1 R a2 - and -NR b -;

R a1 and R a2 are independently selected from hydrogen, deuterium, halogen and C 1-6 alkyl optionally substituted with one or more deuterium or halogen; alternatively, R a1 , R a2 and the carbon atom to which they are commonly attached form an optional C 3-6 cycloalkylene substituted with one or more deuterium or halogen;

R b is selected from hydrogen, deuterium and C 1-6 alkyl optionally substituted with one or more deuterium or halogen;

Further, X is preferably O.
The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound shown in formula I is selected from:
According to the isotopic substitution of the compound according to any one of claims 1-24 or a pharmaceutically acceptable salt thereof, preferably, the isotopic substitution is a deuterium.
A pharmaceutical composition comprising the compound of claims 1-24 or a pharmaceutically acceptable salt thereof, or the isotopic substitution of claim 25, and at least one pharmaceutically acceptable carrier.
Use of the compound of claims 1-24 or a pharmaceutically acceptable salt thereof, or the isotopic substitution of claim 25, or the pharmaceutical composition of claim 26 in the preparation of a medicament for the treatment of a disease associated with P2X3 activity use.
Use of the compound of claims 1-24 or a pharmaceutically acceptable salt thereof, or the isotopic substitution of claim 25, or the pharmaceutical composition of claim 26 in the preparation of a medicament for the treatment of a disease, the disease Selected from pain, urinary tract disease and cough.