Tricyclic glp-1 receptor agonists and uses thereof

Abstract

The present application provides a series of tricyclic GLP-1 receptor agonist compounds, preparation methods and pharmaceutical uses thereof, and the compounds can be used for preparing GLP-1 mediated diseases and related diseases.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to GLP-1 receptor agonist compounds and their preparation methods, as well as the use of said compounds in the preparation of medicaments for treating or preventing GLP-1-mediated diseases and related diseases. Background Technology

[0002] Diabetes mellitus is a chronic, complex disease primarily characterized by disordered glucose metabolism, caused by absolute or relative insulin deficiency or decreased sensitivity of target cells to insulin. It is classified into type 1 and type 2 diabetes mellitus. Type 2 diabetes mellitus is an adult-onset diabetes mellitus, an endocrine disorder primarily characterized by chronic hyperglycemia caused by insulin resistance and / or insulin secretion defects. Type 2 diabetes mellitus accounts for more than 90% of all diabetes cases.

[0003] Currently, the main classes of drugs available for treating type 2 diabetes include: insulin secretagogues, metformin, alpha-glucosidase inhibitors, insulin sensitizers, sodium-glucose cotransporter 2 inhibitors, dipeptidyl peptidase-4 (DPP-4) inhibitors, GLP-1 receptor agonists, insulin, and analogues. Among these, insulin and GLP-1 receptor agonists are among the most effective diabetes treatments. Insulin remains the most widely used diabetes medication globally, with approximately 30-40% of type 2 diabetes patients eventually requiring insulin. GLP-1 inhibitors, such as exenatide, liraglutide, and semaglutide, are suitable for type 2 diabetes patients whose blood sugar cannot be adequately controlled by combined use of metformin, sulfonylureas, and other antibiotics. However, current insulin and GLP-1 inhibitors are primarily peptide drugs and injectable formulations. Even oral semaglutide has several limitations in its use. Therefore, further development of small molecule GLP-1 receptor agonists is still necessary.

[0004] GLP-1 stimulates insulin secretion in a glucose-dependent manner and inhibits glucagon secretion in a glucose-dependent manner, thus posing no risk of hypoglycemia. GLP-1 increases the amount of insulin produced by β-cells, enhancing their responsiveness to glucose. GLP-1 can delay gastric emptying, reducing food intake and thus contributing to weight loss. Furthermore, GLP-1 offers unique cardiovascular benefits. In clinical applications, GLP-1 receptor agonists are positioned as a transitional drug between oral hypoglycemic agents and insulin, and can be used in combination with other medications. They have become the fastest-growing hypoglycemic drug in the past five years and are considered to have the greatest growth potential in the future.

[0005] Other conditions associated with type 2 diabetes include diabetic nephropathy, diabetic eye complications (diabetic retinopathy, diabetes-associated uveitis, diabetic cataracts), diabetic foot, diabetic cardiovascular complications, diabetic cerebrovascular disease, diabetic neuropathy, obesity, and hypertension.

[0006] GLP-1 receptor agonists are highly promising drugs, but most currently available medications are administered via injection. Developing oral small-molecule GLP-1 receptor agonists could improve patient compliance and represents a future trend in GLP-1 receptor agonist development. The known progress in the development of small-molecule GLP-1 receptor agonists is as follows:

[0007] Documents WO2009111700A2 disclose a series of oxadiazathane GLP-1 receptor agonist compounds; WO2010114824A1 discloses a series of substituted azoanthracene derivative GLP-1 receptor agonist compounds; WO2017078352A1 discloses a series of cyclohexene derivative GLP-1 receptor agonist compounds; KR1020180101671A discloses a series of heteroaryl-substituted pyridine [1,2-a] pizimidazole derivative GLP-1 receptor agonist compounds; WO2018056453A1 discloses a series of pyrazolopyridine derivative GLP-1 receptor agonist compounds; and WO2018109607A1 discloses a series of GLP-1 receptor agonist compounds similar to those in this application. Summary of the Invention

[0008] This invention provides a series of compounds as shown in Formula I-2.

[0009]

[0010] and its pharmaceutically acceptable salts, of which

[0011] ------ indicates whether the key exists or does not exist;

[0012] W2 is selected from CH2, CR y ;

[0013] Z1 and Z4 are each independently selected from CH or N;

[0014] Y1 is selected from CH or N;

[0015] Y2 is selected from CH, N, or C;

[0016] Y3 is selected from CH or N;

[0017] R1 is independently selected from hydrogen, oxo, halogen, -CN, -R8, -CO-R8, -CO-NH-R8, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups in R1 and R8 may optionally be independently selected from R x The substituents are substituted 1 to 3 times;

[0018] R2 is selected from -R z -C 1～3 Alkylene-R z -C 0～3 alkylene-amino-R z -SR z -C 0～3 alkylene-carbonyl-R z -C 0～3 alkylene-amide-R z -C 0～3 alkylene-sulfonyl-R z -C 0～3 alkylene-phosphoryl-R z The alkyl, amino, amide, and sulfonyl groups in R2 may optionally be substituted by halogens 1 to 3 times or by R2, provided that the valence allows. w Replaced 0 to 1 times;

[0019] R3 is independently selected from hydrogen, oxo, halogen, -CN, and -C. 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 1～6 Alkoxy, amino, amide, sulfonyl, sulfonylamino, -OH, -C 3～8 Cycloalkyl, 3-8 membered heterocyclic, 6-10 membered aryl, 5-8 membered heteroaryl, wherein R3 may be optionally selected independently from R, provided that the valence allows. y The substituents are substituted 1 to 3 times;

[0020] R4 is independently selected from hydrogen, halogen, and -C. 1～3 Alkyl, -C 1～3 Haloalkyl, -C 1～3 Alkoxy, cyano, hydroxy, amino, amide, sulfonyl, sulfonylamino;

[0021] R5 is independently selected from hydrogen, halogen, hydroxyl, -CN, and -C. 1～3 Alkyl, -C 1～3 Alkoxy, -C 3～6 Cycloalkyl, wherein the alkyl, alkoxy, or cycloalkyl group in R5 may optionally be derived from a halogen, hydroxyl group, or -NR, provided that the valence allows. z -CN, -C 1～3 Alkyl, -C 1～3Alkoxy, -C 1～3 Cycloalkyl substitution 1 to 3 times;

[0022] R6 is selected from -COOH, -CH2COOH, -CH2CH2COOH, and -CH(CH3)COOH, wherein R6 may optionally be substituted by a halogen 1 to 3 times, provided that the oxidation state allows.

[0023] R8 is independently selected from -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 1～6 Alkoxy, C 3～8 Cycloalkyl, 3-8 membered heterocyclic, 6-8 membered aryl, 5-8 membered heteroaryl, wherein the alkyl, alkenyl, ynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups in R8 may optionally be independently selected from R x The substituents are substituted 1 to 3 times;

[0024] n is an integer selected from 0, 1, 2 or 3;

[0025] m is an integer selected from 0, 1, or 2;

[0026] o is an integer selected from 0, 1, 2, 3 or 4;

[0027] p is an integer selected from 0, 1, 2, 3 or 4;

[0028] When o is not 0 and p is not 0, any R4 and R5 can be further cyclized into 5- to 8-membered rings. The resulting rings can be optionally substituted 1 to 3 times with alkyl, haloalkyl, halogen, cyano, oxo, or alkoxy groups, provided that the compound allows it.

[0029] R w Independently selected from -CN, -CH2CN, -C 1～3 Alkyl, -OH, -C 1～3 Alkoxy, amide, sulfonyl, sulfonylamino, -NH2, -NH-C 1～3 Alkyl, wherein the R w The alkyl group in the compound may optionally be converted from C1 to C2, provided that the valence allows. 1～3 Alkyl, C 1～3 Halogenated alkyl, halogen, cyano, C 1～3 Alkoxy substitution 1 to 3 times;

[0030] R x Independently selected from hydrogen, halogen, oxo, C 1～6 Alkoxy, cyano, hydroxy, carboxyl, amino, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C3～6 cycloalkyl, 3-6 membered heterocyclic, 6-8 membered aryl, 5-8 membered heteroaryl, wherein R x The alkyl, alkoxy, cycloalkyl, heterocyclic aryl and heteroaryl groups may optionally be substituted by halogens 1 to 3 times or by hydroxyl groups 0 to 1 time, provided that the valence allows.

[0031] R y Independently selected from hydrogen, halogen, oxo, -C 1～3 Alkoxy, cyano, hydroxy, amino, carboxyl, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 3～6 cycloalkyl, 3- to 6-membered heterocyclic groups, wherein R y Alkyl, alkoxy, cycloalkyl, and heterocyclic groups may optionally be substituted with halogens 1 to 3 times, provided that the valence allows;

[0032] R z Independently selected from hydrogen and C 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 Cycloalkyl, 4-6 membered heterocyclic, 5-6 membered aryl or 5-6 membered heteroaryl, wherein R z Halogens, cyano groups, and C groups can be used optionally, provided the oxidation state allows. 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 The cycloalkyl group or 3- to 6-membered heterocyclic group is substituted 1 to 3 times.

[0033] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically converted into 5- to 8-membered rings; the 5- to 8-membered rings include C 5～6 The rings formed by carbon rings, 5- to 8-membered heterocycles, benzene rings, and 5- to 8-membered heteroaromatic rings may optionally be substituted 1 to 3 times with alkyl, haloalkyl, halogen, cyano, or alkoxy groups, provided that the compound allows.

[0034] Furthermore, the compounds of formula I-2 provided by this invention are tricyclic compounds as shown in formulas I-2-A and I-2-B, obtained by cyclizing R4 and R5:

[0035]

[0036] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically formed into 5- to 8-membered rings, wherein the 5- to 8-membered rings can be selected from... The resulting 5- to 8-membered rings can be arbitrarily composed of C, provided that the oxidation state allows. 1-3 Alkyl, C 1-3 Halogenated alkyl, halogen, cyano, oxo, C 1-3 Alkoxy substitution occurs 1 to 3 times.

[0037] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically formed into a 5- to 8-membered ring, wherein the 5- to 8-membered ring is preferably: The resulting 5- to 8-membered rings can be arbitrarily composed of C, provided that the oxidation state allows. 1-3 Alkyl, C 1-3 Halogenated alkyl, halogen, cyano, oxo, C 1-3 Alkoxy substitution occurs 1 to 3 times.

[0038] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically formed into 5- to 8-membered rings, wherein the 5- to 8-membered rings can be selected from: The resulting 5- to 8-membered rings can be arbitrarily composed of C, provided that the oxidation state allows. 1-3 Alkyl, C 1-3 Halogenated alkyl, halogen, cyano, oxo, C 1-3 Alkoxy substitution occurs 1 to 3 times.

[0039] In one specific implementation, m is preferably 0 or 1;

[0040] In one specific implementation, o is preferably 0, 1, or 2;

[0041] Furthermore, the present invention provides a series of compounds as shown in Formulas I-3.

[0042]

[0043] Furthermore, the compounds of formula I-3 provided by this invention are tricyclic compounds as shown in formulas I-3-A and I-3-B, obtained by cyclizing R4 and R5:

[0044]

[0045] and its pharmaceutically acceptable salts, of which

[0046] ------ indicates whether the key exists or does not exist;

[0047] Y1 is selected from CH or N;

[0048] Y2 is selected from CH, N, or C;

[0049] Y3 is selected from CH or N;

[0050] R1 is independently selected from hydrogen, oxo, halogen, -CN, -R8, -CO-R8, -CO-NH-R8, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups in R1 and R8 may optionally be independently selected from R x The substituents are substituted 1 to 3 times;

[0051] R2 is selected from -R z -C 1～3 Alkylene-R z -C 0～3 alkylene-amino-R z -SR z -C 0～3 alkylene-carbonyl-R z -C 0～3 alkylene-amide-R z -C 0～3 alkylene-sulfonyl-R z -C 0～3 alkylene-phosphoryl-R z The alkyl, amino, amide, and sulfonyl groups in R2 may optionally be substituted by halogens 1 to 3 times or by R2, provided that the valence allows. w Replaced 0 to 1 times;

[0052] R4 is independently selected from hydrogen, halogen, and -C. 1～3 Alkyl, -C 1～3 Haloalkyl, -C 1～3 Alkoxy, cyano, hydroxy, amino, amide, sulfonyl, sulfonylamino;

[0053] R5 is selected from -F, -Cl, -CN, -CH3, -CH2CH3, -CF3, -CHF2, -CH2F, -CH2OH, -OH, -CH2OCH3, -OCH3, -CH2CH2OH, -CH2CH2OCH3, isopropyl, or cyclopropyl.

[0054] R6 is selected from -COOH, -CH2COOH, -CH2CH2COOH, and -CH(CH3)COOH, wherein R6 may optionally be substituted by a halogen 1 to 3 times, provided that the oxidation state allows.

[0055] R8 is independently selected from -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 1～6 Alkoxy, C 3～8Cycloalkyl, 3-8 membered heterocyclic, 6-8 membered aryl, 5-8 membered heteroaryl, wherein the alkyl, alkenyl, ynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups in R8 may optionally be independently selected from R x The substituents are substituted 1 to 3 times;

[0056] n is an integer selected from 0, 1, 2 or 3;

[0057] p is an integer selected from 0, 1, 2, 3 or 4;

[0058] When o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclized into 5- to 8-membered rings. The resulting rings can be optionally substituted 1 to 3 times by alkyl, haloalkyl, halogen, cyano, or alkoxy groups, provided that the compound allows it.

[0059] R w Selected from -CN, -CH2CN, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne group, -OH, -C 1～3 Alkoxy, amide, sulfonyl, sulfonylamino, -NH2, -NH-C 1～3 Alkyl, wherein the R w The alkyl group in the compound may optionally be converted from C1 to C2, provided that the valence allows. 1～3 Alkyl, C 1～3 Halogenated alkyl, halogen, cyano, C 1～3 Alkoxy substitution 1 to 3 times;

[0060] R x Selected from hydrogen, halogen, oxo, C 1～6 Alkoxy, cyano, hydroxy, carboxyl, amino, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 3～6 cycloalkyl, 3-6 membered heterocyclic, 6-8 membered aryl, 5-8 membered heteroaryl, wherein R x The alkyl, alkoxy, cycloalkyl, heterocyclic aryl and heteroaryl groups may optionally be substituted by halogens 1 to 3 times, provided that the valence allows.

[0061] R y Independently selected from hydrogen, halogen, oxo, -C 1～3 Alkoxy, cyano, hydroxy, amino, carboxyl, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 3～6 cycloalkyl, 3- to 6-membered heterocyclic groups, wherein R yAlkyl, alkoxy, cycloalkyl, and heterocyclic groups may optionally be substituted with halogens 1 to 3 times, provided that the valence allows;

[0062] R z Independently selected from hydrogen and C 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 Cycloalkyl, 4-6 membered heterocyclic, 5-6 membered aryl or 5-6 membered heteroaryl, wherein R z Halogens, cyano groups, and C groups can be used optionally, provided the oxidation state allows. 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 The cycloalkyl group or 3- to 6-membered heterocyclic group is substituted 1 to 3 times.

[0063] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically converted into 5- to 8-membered rings; the 5- to 8-membered rings include C 5～6 The rings formed by carbon rings, 5- to 8-membered heterocycles, benzene rings, and 5- to 8-membered heteroaromatic rings may optionally be substituted 1 to 3 times with alkyl, haloalkyl, halogen, cyano, or alkoxy groups, provided that the compound allows.

[0064] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically formed into 5- to 8-membered rings, wherein the 5- to 8-membered rings can be selected from... The resulting 5- to 8-membered rings can be arbitrarily composed of C, provided that the oxidation state allows. 1-3 Alkyl, C 1-3 Halogenated alkyl, halogen, cyano, oxo, C 1-3 Alkoxy substitution occurs 1 to 3 times.

[0065] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically formed into a 5- to 8-membered ring, wherein the 5- to 8-membered ring is preferably: The resulting 5- to 8-membered rings can be arbitrarily composed of C, provided that the oxidation state allows. 1-3 Alkyl, C 1-3 Halogenated alkyl, halogen, cyano, oxo, C 1-3 Alkoxy substitution occurs 1 to 3 times.

[0066] As one specific implementation method, n is selected from 1, 2 or 3, preferably n = 2.

[0067] As one specific implementation, p is selected from 0, 1 or 2, preferably p = 1.

[0068] As a specific implementation, R1 may be further independently selected from -F, -Cl, -CN, -OCH3, -OCH2CH3, -CH3, -CH2CH3, -COCH3, -CONH2, -CF3, -CHF2, -CH2F, -CH2CH2F, -CO-cyclopropyl.

[0069] In one specific implementation, R3 may be further selected from -F, -Cl, -CH3, -OCH3, -NH2, -OH, -CH2CH3, -CH2OH, -NHCH3, -COCH3, -SO2CH3, -OCH2CH3, -CF3, -CHF2, -CH2F, isopropyl, cyclopropyl, and fluorocyclopropyl.

[0070] As a specific implementation, R4 may be further selected from -CN, -CH3, -OH, -CH2OH, -CH2OCH3, -OCH3, -NH2, -NHCH3, -COCH3, and -OCH2CH3.

[0071] As a specific implementation method, R6 can be further preferably -COOH.

[0072] In one specific implementation, R8 is selected from -CH2CH3, -CH3, -CF3, -CHF2, -CH2F, isopropyl, and cyclopropyl.

[0073] As one specific implementation method, the R y It can be further selected from -F, -Cl, methyl, ethyl, trifluoromethyl, difluoromethyl, fluoromethyl, fluoroethyl, methoxy, amino, hydroxy, propyl, isopropyl, cyclopropyl, and cyclobutyl.

[0074] As one specific implementation, the -C(R) of R6 y ) n R in -COOH y It can be connected to C as a main chain and / or a side chain; when R y When connected to C of R6 as a main chain, R y It exists in the form of the corresponding subunit; when R y When connected to C of R6 in a branched manner, R y It exists in the form of the corresponding saturated radical.

[0075] As one specific implementation, the -C(R) of R6 y ) n R in -COOH y When it is methyl, "attached to C in the main chain form" means that... The structures are connected (i.e., R6 is -CH2- at this time), and the connection to C in the form of a branch refers to... The structures are connected (i.e., R6 is -CH3 at this time).

[0076] As one specific implementation method, the R z It can be further selected from: methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy, ethoxy. R z Halogens, cyano groups, and C groups can be used optionally, provided the oxidation state allows. 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 The cycloalkyl group or 3- to 6-membered heterocyclic group is substituted 1 to 3 times.

[0077] In one specific implementation, Y1 is CH or N, preferably Y1 is N;

[0078] In one specific implementation, W1 is O or NH, preferably W1 is O.

[0079] In one specific implementation, -R2 is -R7-R2', where R7 is selected from single bonds, -C 1～3 Alkylene, amino, amide, sulfonyl, sulfonamide.

[0080] As one specific implementation, R2' may be further selected from: methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy, ethoxy, The R2' may be optionally replaced by a halogen 1 to 3 times, or optionally by C, provided that the oxidation state allows. 1～3 Alkyl, C 1～3 Halogenated alkyl, cyano, C 1～3 Alkoxy substitution occurs 0 to 1 time.

[0081] As one specific embodiment, the present invention provides a series of compounds, which are independently selected from one of the following compounds or any combination thereof:

[0082] And its pharmaceutically acceptable salts.

[0083] As one specific embodiment, the present invention provides a series of compounds, which are independently selected from one of the following compounds or any combination thereof:

[0084] And its pharmaceutically acceptable salts.

[0085] Based on the foregoing, the present invention provides a series of compounds as shown in Formula I.

[0086]

[0087] and its pharmaceutically acceptable salts, of which

[0088] W1 is selected from O, S, CH2, and NH;

[0089] W2 is selected from CH2, CR y ;

[0090] Z1, Z2, Z3, and Z4 are each independently selected from CH, N, or C;

[0091] Ring B is selected from aromatic rings or 5- to 6-membered heteroaromatic rings, wherein the heteroaromatic ring is an aromatic ring that is optionally substituted with N atoms 1 to 3 times;

[0092] Ring C is selected from aromatic rings, 4-8 membered heterocyclic rings, 4-10 membered spirocyclic rings, 4-10 membered bridging rings, and 5-7 membered heteroaromatic rings;

[0093] R1 is independently selected from hydrogen, oxo, halogen, -CN, -R8, -CO-R8, -CO-NH-R8, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups in R1 and R8 may optionally be independently selected from R x The substituents are substituted 1 to 3 times;

[0094] R2 is selected from -R z -C 1～3 Alkylene-R z -C 0～3 alkylene-amino-R z -OR z -SR z -C 0～3 alkylene-carbonyl-R z -C 0～3 alkylene-amide-R z -C 0～3 alkylene-sulfonyl-R z -C 0～3 alkylene-phosphoryl-R z The alkyl, amino, amide, and sulfonyl groups in R2 may optionally be substituted by halogens 1 to 3 times or by R, provided that the valence allows. w Replaced 0 to 1 times;

[0095] R3 is independently selected from hydrogen, oxo, halogen, -CN, and -C. 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 1～6Alkoxy, amino, amide, sulfonyl, sulfonylamino, -OH, -C 3～8 Cycloalkyl, 3-8 membered heterocyclic, 6-10 membered aryl, 5-8 membered heteroaryl, wherein R3 may be optionally selected independently from R, provided that the valence allows. y The substituents are substituted 1 to 3 times;

[0096] R4 is independently selected from hydrogen, halogen, and -C. 1～3 Alkyl, -C 1～3 Haloalkyl, -C 1～3 Alkoxy, cyano, hydroxy, amino, amide, sulfonyl, sulfonylamino;

[0097] R5 is independently selected from hydrogen, halogen, hydroxyl, -CN, and -C. 1～3 Alkyl, -C 1～3 Alkoxy, -C 1～3 Cycloalkyl, wherein the alkyl, alkoxy, or cycloalkyl group in R5 may optionally be derived from a halogen, hydroxyl group, or -NR, provided that the valence allows. z -CN, -C 1～3 Alkyl, -C 1～3 Alkoxy, -C 1～3 Cycloalkyl substitution 1 to 3 times;

[0098] R6 is selected from -COOH or -C(R) y ) n -COOH, the -C(R) y ) n -in R y They can be connected to C in the form of a main chain and / or a branch chain; where n is an integer selected from 0, 1, or 2; when n is 2, the two R chains are linked together. y It can be further cyclized into 3- to 8-membered carbon rings or heterocycles;

[0099] R8 is independently selected from -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 1～6 Alkoxy, C 3～8 Cycloalkyl, 3-8 membered heterocyclic, phenyl, 5-8 membered heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups in R8 may optionally be independently selected from R x The substituents are substituted 1 to 3 times;

[0100] n is an integer selected from 0, 1, 2 or 3;

[0101] m is an integer selected from 0, 1, or 2;

[0102] o is an integer selected from 0, 1, 2, 3 or 4;

[0103] p is an integer selected from 0, 1, 2, 3 or 4;

[0104] When m is 2, the two R3s can be further cyclized into 3- to 8-membered carbon rings or heterocycles;

[0105] When m is 1 or 2, R1 and R3 can be further cyclized into 3- to 8-membered carbon rings or heterocycles;

[0106] When o is not 0 and p is not 0, any R4 and R5 can be further cyclized into 5- to 8-membered rings. The resulting rings can be optionally substituted 1 to 3 times with alkyl, haloalkyl, halogen, cyano, oxo, or alkoxy groups, provided that the compound allows it.

[0107] R w Independently selected from -CN, -CH2CN, -C 1～3 Alkyl, -OH, -C 1～3 Alkoxy, amide, sulfonyl, sulfonylamino, -NH2, -NH-C 1～3 Alkyl, wherein the R w The alkyl group in the compound may optionally be converted from C1 to C2, provided that the valence allows. 1～3 Alkyl, C 1～3 Halogenated alkyl, halogen, cyano, C 1～3 Alkoxy substitution 1 to 3 times;

[0108] R x Independently selected from hydrogen, halogen, oxo, C 1～6 Alkoxy, cyano, hydroxy, carboxyl, amino, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 3～6 cycloalkyl, 3-6 membered heterocyclic, 6-8 membered aryl, 5-8 membered heteroaryl, wherein R x The alkyl, alkoxy, cycloalkyl, heterocyclic aryl and heteroaryl groups may optionally be substituted by halogens 1 to 3 times or by hydroxyl groups 0 to 1 time, provided that the valence allows.

[0109] R y Independently selected from hydrogen, halogen, oxo, -C 1～3 Alkoxy, cyano, hydroxy, amino, carboxyl, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 3～6 cycloalkyl, 3- to 6-membered heterocyclic groups, wherein R y Alkyl, alkoxy, cycloalkyl, and heterocyclic groups may optionally be substituted with halogens 1 to 3 times, provided that the valence allows;

[0110] R z Independently selected from hydrogen and C 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 Cycloalkyl, 4-6 membered heterocyclic, 5-6 membered aryl or 5-6 membered heteroaryl, wherein R z Halogens, cyano groups, and C groups can be used optionally, provided the oxidation state allows. 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 The cycloalkyl group or 3- to 6-membered heterocyclic group is substituted 1 to 3 times.

[0111] As one specific implementation, ring B can be further selected from:

[0112]

[0113] As one specific implementation, the ring C can be further selected from:

[0114]

[0115] Furthermore, the present invention provides a series of compounds as shown in Formula I-2,

[0116]

[0117] and its pharmaceutically acceptable salts, of which

[0118] ------ indicates whether the key exists or does not exist;

[0119] W2 is selected from CH2, CR y ;

[0120] Z1 and Z4 are each independently selected from CH or N;

[0121] Y1 is selected from CH or N;

[0122] Y2 is selected from CH, N, or C;

[0123] Y3 is selected from CH or N;

[0124] R1 is independently selected from hydrogen, oxo, halogen, -CN, -R8, -CO-R8, -CO-NH-R8, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups in R1 and R8 may optionally be independently selected from R x The substituents are substituted 1 to 3 times;

[0125] R2 is selected from -R z -C 1～3 Alkylene-R z -C 0～3 alkylene-amino-Rz -SR z -OR z -C 0～3 alkylene-carbonyl-R z -C 0～3 alkylene-amide-R z -C 0～3 alkylene-sulfonyl-R z -C 0～3 alkylene-phosphoryl-R z The alkyl, amino, amide, and sulfonyl groups in R2 may optionally be substituted by halogens 1 to 3 times or by R2, provided that the valence allows. w Replaced 0 to 1 times;

[0126] R3 is independently selected from hydrogen, oxo, halogen, -CN, and -C. 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 1～6 Alkoxy, amino, amide, sulfonyl, sulfonylamino, -OH, -C 3～8 Cycloalkyl, 3-8 membered heterocyclic, 6-10 membered aryl, 5-8 membered heteroaryl, wherein R3 may be optionally selected independently from R, provided that the valence allows. y The substituents are substituted 1 to 3 times;

[0127] R4 is independently selected from hydrogen, halogen, and -C. 1～3 Alkyl, -C 1～3 Haloalkyl, -C 1～3 Alkoxy, cyano, hydroxy, amino, amide, sulfonyl, sulfonylamino;

[0128] R5 is independently selected from hydrogen, halogen, hydroxyl, -CN, and -C. 1～3 Alkyl, -C 1～3 Alkoxy, -C 3～6 Cycloalkyl, wherein the alkyl, alkoxy, or cycloalkyl group in R5 may optionally be derived from a halogen, hydroxyl group, or -NR, provided that the valence allows. z -CN, -C 1～3 Alkyl, -C 1～3 Alkoxy, -C 1～3 Cycloalkyl substitution 1 to 3 times;

[0129] R6 is selected from -COOH, -CH2COOH, -CH2CH2COOH, and -CH(CH3)COOH, wherein R6 may optionally be substituted by a halogen 1 to 3 times, provided that the oxidation state allows.

[0130] R8 is independently selected from -C 1～6 Alkyl, -C 2～6alkenyl, -C 2～6 alkynyl group, -C 1～6 Alkoxy, C 3～8 Cycloalkyl, 3-8 membered heterocyclic, phenyl, 5-8 membered heteroaryl;

[0131] n is an integer selected from 0, 1, 2 or 3;

[0132] m is an integer selected from 0, 1, or 2;

[0133] o is an integer selected from 0, 1, 2, 3 or 4;

[0134] p is an integer selected from 0, 1, 2, 3 or 4;

[0135] When o is not 0 and p is not 0, any R4 and R5 can be further cyclized into 5- to 8-membered rings. The resulting rings can be optionally substituted 1 to 3 times with alkyl, haloalkyl, halogen, cyano, oxo, or alkoxy groups, provided that the compound allows it.

[0136] R w Independently selected from -CN, -CH2CN, -C 1～3 Alkyl, -OH, -C 1～3 Alkoxy, amide, sulfonyl, sulfonylamino, -NH2, -NH-C 1～3 Alkyl, wherein the R w The alkyl group in the compound may optionally be converted from C1 to C2, provided that the valence allows. 1～3 Alkyl, C 1～3 Halogenated alkyl, halogen, cyano, C 1～3 Alkoxy substitution 1 to 3 times;

[0137] R x Independently selected from hydrogen, halogen, oxo, C 1～6 Alkoxy, cyano, hydroxy, carboxyl, amino, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 3～6 cycloalkyl, 3-6 membered heterocyclic, 6-8 membered aryl, 5-8 membered heteroaryl, wherein R x The alkyl, alkoxy, cycloalkyl, heterocyclic aryl and heteroaryl groups may optionally be substituted by halogens 1 to 3 times or by hydroxyl groups 0 to 1 time, provided that the valence allows.

[0138] R y Independently selected from hydrogen, halogen, oxo, -C 1～3 Alkoxy, cyano, hydroxy, amino, carboxyl, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6alkynyl group, -C 3～6 cycloalkyl, 3- to 6-membered heterocyclic groups, wherein R y Alkyl, alkoxy, cycloalkyl, and heterocyclic groups may optionally be substituted with halogens 1 to 3 times, provided that the valence allows;

[0139] R z Independently selected from hydrogen and C 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 Cycloalkyl, 4-6 membered heterocyclic, 5-6 membered aryl or 5-6 membered heteroaryl, wherein R z Halogens, cyano groups, and C groups can be used optionally, provided the oxidation state allows. 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 The cycloalkyl group or 3- to 6-membered heterocyclic group is substituted 1 to 3 times.

[0140] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically converted into 5- to 8-membered rings; the 5- to 8-membered rings include C 5～6 The rings formed by carbon rings, 5- to 8-membered heterocycles, benzene rings, and 5- to 8-membered heteroaromatic rings may optionally be substituted 1 to 3 times with alkyl, haloalkyl, halogen, cyano, or alkoxy groups, provided that the compound allows.

[0141] As one specific embodiment, the compound of formula I provided by the present invention comprises the compound of the following formula:

[0142] Z is independently selected from carbon or nitrogen.

[0143] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically formed into 5- to 8-membered rings, wherein the 5- to 8-membered rings can be selected from: The resulting 5- to 8-membered rings can be arbitrarily composed of C, provided that the oxidation state allows. 1-3 Alkyl, C 1-3 Halogenated alkyl, halogen, cyano, oxo, C 1-3 Alkoxy substitution occurs 1 to 3 times.

[0144] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically formed into 5- to 8-membered rings, wherein the 5- to 8-membered rings can be selected from:

[0145] In one specific implementation, m is preferably 0 or 1;

[0146] In one specific implementation, o is preferably 0, 1, or 2;

[0147] Furthermore, the present invention provides a series of compounds as shown in Formulas I-3.

[0148]

[0149] and its pharmaceutically acceptable salts, of which

[0150] ------ indicates whether the key exists or does not exist;

[0151] Y1 is selected from CH or N;

[0152] Y2 is selected from CH, N, or C;

[0153] Y3 is selected from CH or N;

[0154] R1 is independently selected from hydrogen, oxo, halogen, -CN, -R8, -CO-R8, -CO-NH-R8, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups in R1 and R8 may optionally be independently selected from R x The substituents are substituted 1 to 3 times;

[0155] R2 is selected from -R z -C 1～3 Alkylene-R z -C 0～3 alkylene-amino-R z -SR z -OR z -C 0～3 alkylene-carbonyl-R z -C 0～3 alkylene-amide-R z -C 0～3 alkylene-sulfonyl-R z -C 0～3 alkylene-phosphoryl-R z The alkyl, amino, amide, and sulfonyl groups in R2 may optionally be substituted by halogens 1 to 3 times or by R2, provided that the valence allows. w Replaced 0 to 1 times;

[0156] R4 is independently selected from hydrogen, halogen, and -C. 1～3 Alkyl, -C 1～3 Haloalkyl, -C 1～3 Alkoxy, cyano, hydroxy, amino, amide, sulfonyl, sulfonylamino;

[0157] R5 is selected from -F, -Cl, -CN, -CH3, -CH2CH3, -CF3, -CHF2, -CH2F, -CH2OH, isopropyl, or cyclopropyl.

[0158] R6 is selected from -COOH, -CH2COOH, -CH2CH2COOH, and -CH(CH3)COOH, wherein R6 may optionally be substituted by a halogen 1 to 3 times, provided that the oxidation state allows.

[0159] R8 is independently selected from -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 1～6 Alkoxy, C 3～8 Cycloalkyl, 3-8 membered heterocyclic, 6-8 membered aryl, 5-8 membered heteroaryl;

[0160] n is an integer selected from 0, 1, 2 or 3;

[0161] p is an integer selected from 0, 1, 2, 3 or 4;

[0162] When o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclized into 5- to 8-membered rings. The resulting rings can be optionally substituted 1 to 3 times by alkyl, haloalkyl, halogen, cyano, or alkoxy groups, provided that the compound allows it.

[0163] R w Selected from -CN, -CH2CN, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 Alkyne group, -OH, -C 1～3 Alkoxy, amide, sulfonyl, sulfonylamino, -NH2, -NH-C 1～3 Alkyl, wherein the R w The alkyl group in the compound may optionally be converted from C1 to C2, provided that the valence allows. 1～3 Alkyl, C 1～3 Halogenated alkyl, halogen, cyano, C 1～3 Alkoxy substitution 1 to 3 times;

[0164] R x Selected from hydrogen, halogen, oxo, C 1～6 Alkoxy, cyano, hydroxy, carboxyl, amino, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 3～6 cycloalkyl, 3-6 membered heterocyclic, 6-8 membered aryl, 5-8 membered heteroaryl, wherein R x The alkyl, alkoxy, cycloalkyl, heterocyclic aryl and heteroaryl groups may optionally be substituted by halogens 1 to 3 times, provided that the valence allows.

[0165] R y Independently selected from hydrogen, halogen, oxo, -C 1～3 Alkoxy, cyano, hydroxy, amino, carboxyl, amide, sulfonyl, sulfonylamino, -C 1～6 Alkyl, -C 2～6 alkenyl, -C 2～6 alkynyl group, -C 3～6 cycloalkyl, 3- to 6-membered heterocyclic groups, wherein R y Alkyl, alkoxy, cycloalkyl, and heterocyclic groups may optionally be substituted with halogens 1 to 3 times, provided that the valence allows;

[0166] R z Independently selected from hydrogen and C 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 Cycloalkyl, 4-6 membered heterocyclic, 5-6 membered aryl or 5-6 membered heteroaryl, wherein R z Halogens, cyano groups, and C groups can be used optionally, provided the oxidation state allows. 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 Cycloalkyl groups and 3- to 6-membered heterocyclic groups are substituted 1 to 3 times;

[0167] As a specific implementation, when o is not 0 and p is not 0, any adjacent R4 and R5 can be further cyclically converted into 5- to 8-membered rings; the 5- to 8-membered rings include C 5～6 The rings formed by carbon rings, 5- to 8-membered heterocycles, benzene rings, and 5- to 8-membered heteroaromatic rings may optionally be substituted 1 to 3 times with alkyl, haloalkyl, halogen, cyano, or alkoxy groups, provided that the compound allows.

[0168] As one specific implementation method, n is selected from 1, 2 or 3, preferably n = 2.

[0169] As one specific implementation, p is selected from 0, 1 or 2, preferably p = 1.

[0170] As a specific implementation, R1 can be further independently selected from -F, -Cl, -CN, -OCH3, -OCH2CH3, -CH3, -CH2CH3, -COCH3, -CONH2, -CF3, -CHF2, -CH2F, and -CH2CH2F.

[0171] In one specific implementation, R3 may be further selected from -F, -Cl, -CH3, -OCH3, -NH2, -OH, -CH2CH3, -CH2OH, -NHCH3, -COCH3, -SO2CH3, -OCH2CH3, -CF3, -CHF2, -CH2F, isopropyl, cyclopropyl, and fluorocyclopropyl.

[0172] As one specific implementation method, the R y It can be further selected from -F, -Cl, methyl, ethyl, trifluoromethyl, difluoromethyl, fluoromethyl, fluoroethyl, methoxy, amino, hydroxy, propyl, isopropyl, cyclopropyl, and cyclobutyl.

[0173] As one specific implementation, the -C(R) of R6 y ) n R in -COOH y It can be connected to C as a main chain and / or a side chain; when R y When connected to C of R6 as a main chain, R y It exists in the form of the corresponding subunit; when R y When connected to C of R6 in a branched manner, R y It exists in the form of the corresponding saturated radical.

[0174] As one specific implementation, the -C(R) of R6 y ) n R in -COOH y When it is methyl, "attached to C in the main chain form" means that... The structures are connected (i.e., R6 is -CH2- at this time), and the connection to C in the form of a branch refers to... The structures are connected (i.e., R6 is -CH3 at this time).

[0175] As one specific implementation method, the R z It can be further selected from: methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy, ethoxy. R z Halogens, cyano groups, and C groups can be used optionally, provided the oxidation state allows. 1～3 Alkyl, C 1～3 Alkoxy, C 3～6 The cycloalkyl group or 3- to 6-membered heterocyclic group is substituted 1 to 3 times.

[0176] In one specific implementation, Y1 is CH or N, preferably Y1 is N;

[0177] In one specific implementation, W1 is O or NH, preferably W1 is O.

[0178] In one specific implementation, -R2 is -R7-R2', where R7 is selected from single bonds, -C 1～3 Alkylene, amino, amide, sulfonyl, sulfonamide.

[0179] As one specific implementation, R2' may be further selected from: methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy, ethoxy, The R2' may be optionally replaced by a halogen 1 to 3 times, or optionally by C, provided that the oxidation state allows. 1～3 Alkyl, C 1～3 Halogenated alkyl, cyano, C 1～3 Alkoxy substitution occurs 0 to 1 time.

[0180] As one specific embodiment, the present invention provides a series of compounds, which are independently selected from one of the following compounds or any combination thereof:

[0181]

[0182] And its pharmaceutically acceptable salts.

[0183] As one specific embodiment, the present invention provides a series of compounds, which are independently selected from one of the following compounds or any combination thereof:

[0184] And its pharmaceutically acceptable salts.

[0185] The compounds provided by this invention and their pharmaceutically acceptable salts can be used alone or in combination with at least one other therapeutic agent in treatment.

[0186] The present invention provides a pharmaceutical composition comprising a compound of formula I and a pharmaceutically acceptable salt thereof, and one or more other therapeutically active ingredients.

[0187] The present invention also provides a pharmaceutical preparation comprising a compound of formula I and a pharmaceutically acceptable salt thereof, and one or more pharmaceutical carriers; the pharmaceutical preparation is any clinically acceptable dosage form.

[0188] The compounds and pharmaceutically acceptable salts provided by this invention can be formulated into solid dosage forms, such as capsules, tablets, pills, lozenges, sugar-coated tablets, granules, powders, ointments, creams, drops, etc.; the compounds and pharmaceutically acceptable salts provided by this invention can be formulated into liquid dosage forms, such as elixirs, syrups, emulsions, dispersants, suspensions, solutions, sprays, etc.

[0189] The pharmaceutical carriers and / or pharmaceutical diluents that can be used in the pharmaceutical compositions or pharmaceutical formulations of the present invention can be any conventional carriers and / or diluents in the field of pharmaceutical formulations.

[0190] The pharmaceutically acceptable salts described in this invention include acid-value salts and basic salts.

[0191] The compounds and pharmaceutically acceptable salts provided by this invention may exist in chiral forms, i.e., S-configuration or R-configuration. The compounds and pharmaceutically acceptable salts provided by this invention may also exist in achiral forms. When the compounds described in this invention are illustrated with one configuration, it also indicates the disclosure of another configuration or achiral form.

[0192] The compounds described in this invention include stereoisomers of the compounds. The stereoisomers described in this invention refer to the following: when a compound contains an asymmetric carbon atom, it produces enantiomers; when a compound contains a carbon-carbon double bond or a cyclic structure, it produces cis-trans isomers; when a compound contains a ketone or oxime, it produces tautomers. As a specific embodiment, the stereoisomers described in this invention include, but are not limited to: enantiomers, diastereomers, racemic isomers, cis-trans isomers, tautomers, geometric isomers, epimers, and mixtures thereof.

[0193] The pharmaceutically acceptable salts described in this invention can exist in both non-solventized and solvated forms.

[0194] The present invention also provides the use of compounds as shown in Formula I and pharmaceutically acceptable salts thereof in the preparation of medicaments for the treatment and / or metabolic-related diseases. The metabolic-related diseases include GLP-1-mediated diseases and related diseases, including but not limited to: diabetes, hyperglycemia, insulin resistance, glucose intolerance, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, adipocyte dysfunction, obesity, dyslipidemia, hyperinsulinemia, etc.; wherein the diabetes includes, but is not limited to, type 1 diabetes mellitus (T1D) and / or type 2 diabetes mellitus (T2DM), idiopathic T1D, early-onset T2D, latent autoimmune diabetes mellitus, juvenile atypical diabetes mellitus, gestational diabetes mellitus, etc.

[0195] The present invention also provides a method for treating a disease, comprising administering to a patient in need a therapeutically effective amount of a compound as shown in Formula I and a pharmaceutically acceptable salt thereof, wherein the disease is a GLP-1 mediated disease or related disease; the disease includes, but is not limited to: diabetes, hyperglycemia, insulin resistance, glucose intolerance, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, adipocyte dysfunction, obesity, dyslipidemia, hyperinsulinemia, etc.; wherein the diabetes includes, but is not limited to, type 1 diabetes mellitus (T1D) and / or type 2 diabetes mellitus (T2DM), idiopathic T1D, early-onset T2D, latent autoimmune diabetes mellitus, juvenile atypical diabetes mellitus, gestational diabetes mellitus, etc.

[0196] The compounds shown in Formula I and their pharmaceutically acceptable salts provided by this invention have excellent GLP-1 receptor agonist activity and can treat and / or prevent GLP-1-mediated diseases and related diseases.

[0197] The present invention also provides the use of the above-described compounds or pharmaceutically acceptable salts thereof in the preparation of GLP-1 receptor agonist-related drugs.

[0198] In some embodiments of the present invention, the GLP-1 receptor agonist-related drugs are used to treat type II diabetes, type I diabetes, and obesity.

[0199] The compounds described in this invention are named according to their chemical structural formulas. If the name of the compound does not match the chemical structural formula when referring to the same compound, the chemical structural formula shall prevail.

[0200] Unless otherwise stated, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. However, for a better understanding of this invention, definitions of some terms are provided below. When the definitions and interpretations of terms provided in this invention differ from the meanings commonly understood by those skilled in the art, the definitions and interpretations provided in this invention shall prevail.

[0201] The compounds and pharmaceutically acceptable salts provided by this invention may exist in chiral forms, i.e., S-configuration or R-configuration. The compounds and pharmaceutically acceptable salts provided by this invention may also exist in achiral forms. When the compounds described in this invention are illustrated with one configuration, it also indicates the disclosure of another configuration or achiral form.

[0202] The compounds described in this invention include stereoisomers of the compounds. The stereoisomers described in this invention refer to the following: when a compound as shown in Formula I contains an asymmetric carbon atom, it produces an enantiomer; when the compound contains a carbon-carbon double bond or a cyclic structure, it produces a cis-trans isomer; when the compound contains a ketone or oxime, it produces a tautomer. As a specific embodiment, the stereoisomers described in this invention include, but are not limited to: enantiomers, diastereomers, racemic isomers, cis-trans isomers, tautomers, geometric isomers, epimers, and mixtures thereof.

[0203] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0204] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0205] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0206] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0207] Unless otherwise stated, "(+)" indicates right-handed rotation, "(-)" indicates left-handed rotation, and "(±)" indicates racemic rotation.

[0208] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key This indicates that the center of the solid is an absolute configuration, but it is uncertain whether it is a wedge-shaped solid line key. or wedge-shaped dashed key Use wavy lines Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key Or straight dashed key

[0209] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated, and the auxiliary group is cleaved to provide the desired enantiomer in pure form. Alternatively, when the molecule contains a basic functional group (such as an amine) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase and optionally combined with chemical derivatization (e.g., from amines to form carbamates).

[0210] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0211] The term "pharmaceutically acceptable" in this invention means that, with respect to those compounds, materials, compositions and / or dosage forms, they are suitable for use in contact with human and animal tissues, within the limits of reliable medical judgment, without excessive toxicity, irritation, allergic reactions or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0212] The term "pharmaceutically acceptable salt" in this invention refers to a salt of a compound of this invention, prepared by reacting a compound having specific substituents discovered in this invention with a relatively non-toxic acid or base. When a compound of this invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting such a compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When a compound of this invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting such a compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; salts of amino acids (such as arginine); and salts of organic acids such as glucuronic acid. Certain compounds of the present invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0213] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0214] The term "optional" or "optionally" in this invention refers to an event or condition that may, but is not required, to occur as described below, and the description includes both cases where said event or condition occurs and cases where said event or condition does not occur.

[0215] The term "substituted" in this invention means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted. Unless otherwise specified, the type and number of substituents can be arbitrary on a chemically feasible basis.

[0216] The term "optionally replaced" in this invention refers to both "replaced" and "not replaced" scenarios.

[0217] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0218] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0219] When the number of a substituent is 0, it means that the substituent does not exist. For example, -A-(R)0 means that the structure is actually -A.

[0220] When a substituent is vacant, it means that the substituent does not exist. For example, if X is vacant in AX, it means that the structure is actually A.

[0221] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0222] When a substituent can be cross-bonded to two or more atoms on a ring, this substituent can bond with any atom on that ring, for example, a structural unit. This indicates that the substituent R can be substituted at any position on the cyclohexyl or cyclohexadiene. When the listed substituents do not specify which atom they are attached to the substituted group, such substituents can be bonded to any of their atoms. For example, a pyridyl group as a substituent can be attached to the substituted group through any carbon atom on the pyridine ring.

[0223] When the listed linking groups do not specify their linking direction, the linking direction is arbitrary, for example, The linker group L is -MW-. In this case, -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form a ring. Alternatively, rings A and B can be connected in the opposite direction to the left-to-right reading order to form a ring. The combination of linking groups, substituents, and / or their variants is permitted only if such a combination produces a stable compound.

[0224] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of that group can be connected to other groups by chemical bonds. When the chemical bond connection is non-directional and the connectable site contains H atoms, the number of H atoms at that site will decrease accordingly with the number of chemical bonds connected, resulting in a group with a corresponding valence. The chemical bonds connecting the site to other groups can be straight solid line bonds. Straight dashed key or wavy line For example, a straight solid line bond in -OCH3 indicates that the oxygen atom in that group is connected to other groups; The straight dashed bond in the diagram indicates that the group is connected to other groups through both ends of the nitrogen atom in the group; The wavy lines in the text indicate that the phenyl group is connected to other groups through the carbon atoms at positions 1 and 2 of the phenyl group. This indicates that any connectable site on the piperidinyl group can be linked to other groups via a single chemical bond, including at least... Even if H atoms are drawn on -N- in these four connection methods, Still includes In this type of linkage, when a chemical bond is attached, the number of hydrogen atoms at that site is reduced by one, resulting in a monovalent piperidinyl group.

[0225] Unless otherwise specified, the number of atoms in a ring is usually defined as the elemental number of the ring. For example, a “5-7 elemental ring” refers to a “ring” with 5-7 atoms arranged around it.

[0226] In this invention, the term "halogen atom" refers to fluorine, chlorine, bromine, iodine, etc. Preferably, the halogen atom used as a substituent in the aryl group of this invention is a fluorine or chlorine atom. Preferably, the halogen atom used as a substituent in the alkyl group of this invention is a fluorine or chlorine atom. C having a halogen atom as a substituent... 1-6 Alkyl groups include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, pentafluoroethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 2-chloroethyl, heptafluoropropyl, 3,3,3-trifluoropropyl, 2,3-dichloropropyl, 1-fluoro-3-bromopropyl, 4-bromobutyl, 3,3,3,4,4-pentafluorobutyl, 4,4-dichlorobutyl, 5-iodopentyl, 5,5-difluoropentyl, 6-chlorohexyl, and 6,6,6-trifluorohexyl.

[0227] The term "C" in this invention 1～6"Alkyl" refers to a straight-chain or branched alkyl group having 1 to 6 carbons, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, 1-methylpropyl, n-pentyl, isopentyl, 2-methylbutyl, 1,1-dimethylpropyl, 1-ethylpropyl, n-hexyl, 4-methylpentyl, and 2-ethylbutyl.

[0228] The term "C" in this invention 1～6 "Alkoxy" refers to the carbon group. 1-6 Alkyl-O-, including but not limited to methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, 1-methylpropoxy, n-pentyloxy, isopentyloxy, 2-methylbutoxy, 1,1-dimethylpropoxy, 1-ethylpropoxy, n-hexyloxy, 4-methylpentyloxy, and 2-ethylbutoxy.

[0229] In this invention, the term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group having a conjugated π-electron system, preferably a 6- to 10-membered ring, such as phenyl and naphthyl, more preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, including benzo3- to 8-membered cycloalkyl and benzo3- to 8-membered heterocyclic groups, wherein the heterocyclic group is a heterocyclic group containing 1-3 nitrogen, oxygen, and sulfur atoms; or may further include a three-membered nitrogen-containing fused ring containing a benzene ring.

[0230] The term "heteroaryl" in this invention refers to a heteroaryl system comprising 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. The heteroaryl group is preferably 5 to 10-membered, more preferably 5- or 6-membered, such as imidazolyl, furanyl, thiophene, thiazolyl, pyrazolyl, oxazolyl, pyrroleyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably triazolyl, thiophene, imidazolyl, pyrazolyl, oxazolyl, pyrimidinyl, or thiazolyl. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, including but not limited to:

[0231] The heteroaryl group can be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamine, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl, or carboxylic acid ester group.

[0232] Unless otherwise specified, the terms "5-6-membered heteroaryl" and "5-6-membered heteroaryl" in this invention are used interchangeably. The term "5-6-membered heteroaryl" refers to a monocyclic group consisting of 5 to 6 ring atoms with a conjugated π-electron system, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). p (where p is 1 or 2). The 5-6 membered heteroaryl group can be attached to the rest of the molecule via a heteroatom or a carbon atom. The 5-6 membered heteroaryl group includes both 5-membered and 6-membered heteroaryl groups. Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrole (including N-pyrrole, 2-pyrrole, and 3-pyrrole), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazole (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl), and triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl). (and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isooxazolyl, 4-isooxazolyl and 5-isooxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

[0233] The term "alkoxy" in this invention refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), wherein alkyl is defined as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexoxy. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamine, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl, or carboxylic acid ester groups.

[0234] The term "halogenated alkyl" in this invention refers to an alkyl group that has been substituted with one or more halogens.

[0235] The term "3- to 8-membered heterocyclic group" in this invention refers to a non-aromatic cyclic group comprising one or more heteroatoms selected from nitrogen, oxygen, and sulfur atoms, and may be fully saturated or partially unsaturated. The ring may be a 3- to 8-membered monocyclic, bicyclic, or spirocyclic group. Examples include, but are not limited to, oxacyclobutane, aziranebutane, piperazine, piperidinyl, morpholinyl, thiomorpholinyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, thiohexane, oxacyclohexane, thiazoxane, dihydroindolyl, isodihydroindolyl, tetrahydrodihydroindolyl, quininecycloyl, and azirzoyl.

[0236] The heterocyclic ring may be fused to an aryl, heteroaryl, or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, and non-limiting examples include:

[0237] The term "C" in this invention 3～8 "Cycloalkyl" refers to a monovalent group obtained by removing any single hydrogen atom from a cyclic saturated aliphatic hydrocarbon having 3 to 8 carbons; that is, a cycloalkyl group with 3 to 8 carbons. Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. When two groups together form a C12 group... 3～8 When cycloalkyl groups are formed, the resulting group can be divalent, such as cyclopropane-1,1-diyl, cyclobutane-1,1-diyl, cyclopentane-1,1-diyl, cyclohexane-1,1-diyl, cycloheptane-1,1-diyl, and cyclooctane-1,1-diyl. Furthermore, the cycloalkane ring, carbide ring, and cyclohydrocarbon in cycloalkyl groups can be cross-linked rings.

[0238] The term "fused ring" in this invention refers to a 5- to 20-membered all-carbon polycyclic group, wherein each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, and one or more rings may contain one or more double bonds, but no ring has a fully conjugated π-electron system. Fused rings are preferably 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of constituent rings, fused rings can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused alkyl groups, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl groups. These include, but are not limited to:

[0239] The carbon atom in a fused ring can be optionally replaced by a heteroatom of O, S, or N, which also includes "fused heterocycles".

[0240] The term "fused heterocycle" in this invention refers to a 5- to 20-membered polycyclic heterocyclic group, wherein each ring in the system shares an adjacent pair of atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a fully conjugated π-electron system, and wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O).t (Where t is an integer from 0 to 2) heteroatoms, with the remaining ring atoms being carbon. The fused heterocycle is preferably 6 to 14-membered, more preferably 7 to 10-membered. Depending on the number of rings, the fused heterocycle can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocycle groups, preferably bicyclic or tricyclic, more preferably 5-membered / 3-membered, 5-membered / 4-membered, or 5-membered / 5-membered bicyclic fused heterocycle groups. Fused heterocycles include, but are not limited to:

[0241] The term "bridged ring" in this invention refers to a 5- to 20-membered all-carbon polycyclic group, wherein any two rings share two non-directly connected carbon atoms. The bridged ring may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Bridged rings are preferably 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of constituent rings, they can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged ring groups, preferably bicyclic, tricyclic, or tetracyclic, more preferably bicyclic or tricyclic. Bridged rings include, but are not limited to:

[0242]

[0243] The carbon atom in the bridged ring can be optionally replaced by heteroatoms of O, S, or N, which also includes "bridged heterocycles".

[0244] The term "bridged heterocycle" in this invention refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two non-directly connected atoms. The bridged heterocycle may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. One or more ring atoms are selected from nitrogen, oxygen, or S(O). m (Where m is an integer from 0 to 2) heteroatoms, with the remaining ring atoms being carbon. The bridging heterocycle is preferably 6 to 14-membered, more preferably 7 to 10-membered. Depending on the number of rings, bridging heterocycles can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridging heterocycles, preferably bicyclic, tricyclic, or tetracyclic, more preferably bicyclic or tricyclic. Bridging heterocycles include, but are not limited to:

[0245]

[0246] The term "spirocyclic" in this invention refers to a 5- to 20-membered polycyclic group, wherein the monocyclic rings share a carbon atom (called a spiro atom), and the spirocyclic ring may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Spirocyclic rings are preferably 6- to 14-membered, more preferably 7- to 10-membered. Spirocyclic alkyl groups are classified into monospirocyclic alkyl groups, bispirocyclic alkyl groups, or polyspirocyclic alkyl groups according to the number of shared spiro atoms between the rings, preferably monospirocyclic alkyl groups and bispirocyclic alkyl groups. More preferably, 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospirocyclic alkyl groups are included, but are not limited to:

[0247] The carbon atom in a spiroring can be optionally replaced by heteroatoms of O, S, or N, which also includes "spiroheterorings".

[0248] In this invention, the term "spiroheterocycle" refers to a 5- to 20-membered polycyclic heterocyclic group, wherein the monocyclic rings share a common atom (called a spiro atom), and one or more ring atoms are selected from nitrogen, oxygen, or S(O). m (Where m is an integer from 0 to 2) heteroatoms, with the remaining ring atoms being carbon. Spiroheterocyclic rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Spiroheterocyclic rings are preferably 6 to 14-membered, more preferably 7 to 10-membered. Spiroheterocyclic groups are classified into monospirocyclic, bispirocyclic, or multispirocyclic groups based on the number of shared spiroatoms between rings, with monospirocyclic and bispirocyclic groups being preferred. More preferably, 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospirocyclic groups are preferred. These include, but are not limited to:

[0249] The compounds of the present invention can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples of the present invention. Detailed Implementation

[0250] The present invention will be further described in detail below with reference to specific embodiments. The following embodiments are used to understand the methods and core ideas of the present invention. For those skilled in the art, any possible changes or substitutions made without departing from the concept of the present invention are within the protection scope of the present invention. Experimental methods in the embodiments of the present invention that do not specify specific conditions are generally under conventional conditions or according to the conditions recommended by the raw material or product manufacturer; reagents whose source is not specified are generally commercially available conventional reagents.

[0251] Experiment 1 - Compound Identification and Characterization

[0252] The present invention 1 1H NMR spectra were obtained using a Bruker instrument (400 MHz), and chemical shifts are expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. 1 H NMR representation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublet, dt = doublet of triplet. If the coupling constant is provided, the unit is Hz.

[0253] The mass spectrometry of this invention is obtained by LC / MS, and the ionization method can be ESI or APCI.

[0254] Experimental results:

[0255] Compound 1

[0256]

[0257] 1 H NMR (400MHz, CD3OD) δ = 8.32 (s, 1H), 7.97 (d, J = 7.6Hz, 1H), 7.75–7.61 (m, 2H), 7.59–7.48 (m, 2H), 7.29 (d d,J=8.4,1.7Hz,1H),6.28(dd,J=8.4,2.0Hz,1H),5.54–5.36(m,2H),5.28–5.17(m,1H),4.71–4.61(m,3 H),4.44(dt,J=9.1,5.9Hz,1H),4.33–4.21(m,1H),4.07(d,J=13.7Hz,1H),4.02–3.80(m,2H),3.37(d,J =11.3Hz,1H),3.25(d,J=6.3Hz,3H),3.04–2.69(m,3H),2.51(dd,J=17.8,9.7Hz,1H),2.38–2.13(m,2H).

[0258] Compound 2

[0259]

[0260] 1 H-NMR (400MHz, MeOD) δ8.17(s,1H),7.94(d,J=8.5Hz,1H),7.57(dddd,J=8.9,8.0,6.6,5.3Hz,4H),6.94( dd,J=8.3,0.7Hz,1H),6.07(dd,J=8.3,1.7Hz,1H),5.38(s,2H),5.27(dd,J=7.0,3.3Hz,1H),4.81(d,J=6 .6Hz,1H),4.75–4.58(m,2H),4.51–4.40(m,1H),4.37–4.28(m,1H),4.19–3.77(m,4H),3.34(d,J=11.7Hz ,1H),3.03–2.69(m,4H),2.52(dd,J=17.0,10.1Hz,1H),2.32–2.20(m,1H),2.00(dt,J=16.1,10.8Hz,1H).

[0261] Compounds 2-P1 & 2-P2

[0262]

[0263] 1 H-NMR (400 MHz, MeOD) δ 8.18 (s, 1H), 7.94 (d, J = 8.5 Hz, 1H), 7.67–7.51 (m, 4H), 6.94 (d, J = 8.3 Hz, 1H), 6.07 (d, J = 8.3 Hz, 1H), 5.38 (s, 2H), 5.30–5.22 (m, 1H), 4.92–4.86 (m, 2H), 4.65 (ddd, J = 21.8, 14.6, 5.2 Hz, 2H), 4.47 (dt, J = 9.1, 6.0 Hz, 1H), 4.34 (d, J = 12.8 Hz, 1H), 4.14–4.02 (m, 2H), 3.93–3.80 (m, 2H), 2.97 (d, J = 11.4 Hz, 1H), 2.86–2.75 (m, 3H), 2.57–2.48 (m, 1H), 2.29 (dd, J = 11.6, 8.7 Hz, 1H), 1.98 (t, J = 10.8 Hz, 1H). 1 H-NMR (400 MHz, MeOD) δ 8.41 (s, 1H), 8.31 (s, 1H), 7.98 (d, J = 8.5 Hz, 1H), 7.64 (dd, J = 16.5, 8.1 Hz, 2H), 7.57–7.42 (m, 2H), 6.94 (d, J = 8.3 Hz, 1H), 6.08 (d, J = 8.3 Hz, 1H), 5.31 (dd, J = 45.6, 3.9 Hz, 3H), 4.81 (s, 1H), 4.76–4.58 (m, 2H), 4.44 (dt, J = 9.1, 5.9 Hz, 1H), 4.31 (d, J = 12.5 Hz, 1H), 4.15 (dd, J = 10.9, 2.9 Hz, 1H), 3.92 (ddd, J = 19.0, 15.7, 10.9 Hz, 3H), 3.49–3.34 (m, 1H), 2.99–2.67 (m, 4H), 2.56–2.44 (m, 1H), 2.33–2.18 (m, 1H), 2.04 (t, J = 10.8 Hz, 1H).

[0264] Compounds 3-P1 & 3-P2

[0265]

[0266] 1¹H NMR (400 MHz, MeOD) δ 8.08 (s, 1H), 7.85 (d, J = 8.5 Hz, 1H), 7.49 (d, J = 8.4 Hz, 1H), 7.33 (t, J = 8.2 Hz, 1H), 7.12–6.99 (m, 2H), 6.82 (d, J = 8.3 Hz, 1H), 5.92 (d, J = 8.3 Hz, 1H), 5.25–5.13 (m, 3H), 4.72 (d, J = 6.6 Hz, 1H), 4.62 (dd, J = 15.3, 2.8 Hz, 1H), 4.52 (d, J = 6.0 Hz, 1H), 4.38–4.25 (m, 2H), 4.05 (dd, J = 10.9, 2.8 Hz, 1H), 3.89 (s, 2H), 3.76 (dd, J = 10.8, 8.2 Hz, 1H), 3.27 (d, J = 2.5 Hz, 1H), 2.80 (d, J = 11.0 Hz, 2H), 2.69 (dd, J = 9.1, 2.9 Hz, 2H), 2.42 (s, 1H), 2.17 (d, J = 2.9 Hz, 1H), 1.93 (t, J = 10.8 Hz, 1H)

[0267] 1 ¹H NMR (400 MHz, CDCl₃) δ 8.25 (s, 1H), 8.07 (d, J = 8.4 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.40 (t, J = 8.0 Hz, 1H), 7.14–7.04 (m, 2H), 6.93 (d, J = 8.3 Hz, 1H), 6.06 (d, J = 8.3 Hz, 1H), 5.28 (dd, J = 21.6, 13.2 Hz, 3H), 4.78 (dd, J = 15.1, 6.1 Hz, 1H), 4.69–4.59 (m, 2H), 4.48–4.39 (m, 2H), 4.13–3.99 (m, 3H), 3.91–3.82 (m, 1H), 3.37 (s, 1H), 3.01 (d, J = 10.3 Hz, 1H), 2.87–2.70 (m, 3H), 2.46 (dd, J = 21.9, 10.8 Hz, 2H), 2.06 (t, J = 10.8 Hz, 1H) Compound 4-P1 & 4-P2

[0268]

[0269] 1¹H NMR (400 MHz, MeOD) δ 8.20 (s, 1H), 7.95 (dd, J = 8.4, 1.4 Hz, 1H), 7.65–7.52 (m, 4H), 7.38 (d, J = 8.0 Hz, 1H), 6.29 (d, J = 7.9 Hz, 1H), 5.45 (s, 2H), 5.28 (td, J = 6.9, 4.1 Hz, 1H), 4.83 (s, 1H), 4.71 (dd, J = 15.3, 2.9 Hz, 1H), 4.64 (dd, J = 13.1, 2.8 Hz, 2H), 4.56–4.41 (m, 2H), 3.93 (dt, J = 13.1, 12.0 Hz, 3H), 3.81 (dd, J = 12.9, 2.8 Hz, 1H), 3.71 (dd, J = 12.9, 6.3 Hz, 1H), 3.48–3.41 (m, 1H), 3.23 (t, J = 10.7 Hz, 1H), 2.81–2.73 (m, 3H), 2.56–2.46 (m, 1H), 2.36 (dd, J = 11.0, 8.4 Hz, 1H), 2.23 (t, J = 10.5 Hz, 1H).

[0270] 1 ¹H NMR (400 MHz, MeOD) δ 8.30 (s, 1H), 7.96 (d, J = 8.0 Hz, 1H), 7.67–7.52 (m, 4H), 7.38 (d, J = 8.0 Hz, 1H), 6.30 (d, J = 7.9 Hz, 1H), 5.46 (s, 2H), 5.25 (d, J = 4.8 Hz, 1H), 4.74–4.68 (m, 1H), 4.63 (t, J = 9.0 Hz, 2H), 4.56–4.43 (m, 2H), 4.01 (d, J = 13.7 Hz, 1H), 3.89 (t, J = 11.9 Hz, 2H), 3.79 (dd, J = 12.9, 2.8 Hz, 1H), 3.72–3.63 (m, 1H), 3.41 (s, 1H), 3.24 (s, 2H), 2.82 (t, J = 11.4 Hz, 2H), 2.69 (d, J = 10.6 Hz, 1H), 2.53 (s, 1H), 2.40 (s, 1H), 2.21 (t, J = 10.4 Hz, 1H).

[0271] Compounds 5-P1 & 5-P2

[0272]

[0273] 1¹H NMR (400 MHz, MeOD) δ 8.27 (s, 1H), 7.96 (d, J=8.0 Hz, 1H), 7.67–7.60 (m, 2H), 7.59–7.46 (m, 2H), 7.11 (d, J=8.3 Hz, 1H), 6.23 (d, J=8.3 Hz, 1H), 5.38 (s, 2H), 5.32–5.24 (m, 1H), 4.75 (dd, J=15.4, 2.8 Hz, 2H), 4.66–4.59 (m, 1H), 4.44 (dd, J=5.9, 3.2 Hz, 1H), 4.16–4.02 (m, 2H), 3.96 (s, 2H), 3.72 (d, J=13.1 Hz, 1H), 3.56 (s, 1H), 3.24 (s, 1H), 2.74 (d, J=10.7 Hz, 3H), 2.44 (d, J=2.5 Hz, 2H), 2.24 (d, J=9.5 Hz, 1H), 2.08–1.97 (m, 1H), 1.93 (s, 1H).

[0274] 1 ¹H NMR (400 MHz, MeOD) δ 8.30 (s, 1H), 7.96 (d, J=8.4 Hz, 1H), 7.67–7.60 (m, 2H), 7.55 (t, J=8.4 Hz, 2H), 7.11 (d, J=8.3 Hz, 1H), 6.23 (d, J=8.3 Hz, 1H), 5.39 (s, 2H), 5.30–5.23 (m, 1H), 4.75–4.55 (m, 3H), 4.49 (dd, J=6.0, 3.2 Hz, 1H), 4.13–4.01 (m, 3H), 3.87 (d, J=13.6 Hz, 1H), 3.73 (d, J=13.1 Hz, 1H), 3.54 (s, 1H), 3.26 (s, 1H), 2.84–2.75 (m, 2H), 2.67 (d, J=9.4 Hz, 1H), 2.50 (d, J=25.1 Hz, 2H), 2.22 (d, J=11.0 Hz, 1H), 2.07–1.96 (m, 1H), 1.93 (s, 1H).

[0275] Compound 6

[0276]

[0277] 1¹H NMR (400 MHz, MeOD) δ 8.17 (s, 1H), 7.95 (dd, J = 8.4, 1.5 Hz, 1H), 7.69–7.52 (m, 5H), 6.75 (d, J = 8.2 Hz, 1H), 5.54 (s, 2H), 5.26 (d, J = 4.2 Hz, 2H), 4.70 (dd, J = 15.4, 3.1 Hz, 1H), 4.61 (dd, J = 13.7, 7.8 Hz, 1H), 4.45 (dt, J = 9.1, 5.9 Hz, 1H), 4.35 (s, 2H), 4.10 (dd, J = 34.7, 13.6 Hz, 2H), 3.87 (s, 2H), 3.48 (d, J = 1.7 Hz, 1H), 3.24–3.23 (m, 1H), 2.85–2.71 (m, 5H), 2.53 (d, J = 8.8 Hz, 1H).

[0278] Compound 7

[0279]

[0280] 1 ¹H NMR (400 MHz, CDCl₃) δ 8.19 (q, J = 8.2 Hz, 2H), 7.61 (t, J = 7.5 Hz, 1H), 7.52 (d, J = 8.3 Hz, 1H), 7.44 (d, J = 7.8 Hz, 1H), 7.37 (d, J = 9.1 Hz, 1H), 6.71 (d, J = 8.2 Hz, 1H), 5.51 (s, 2H), 5.25 (s, 1H), 4.93–4.85 (m, 1H), 4.79 (d, J = 12.9 Hz, 1H), 4.61 (d, J = 7.1 Hz, 1H), 4.35 (s, 3H), 4.20 (s, 2H), 3.87 (s, 2H), 3.28 (s, 2H), 2.84 (d, J = 5.4 Hz, 2H), 2.74 (s, 3H), 2.46 (s, 1H).

[0281] Compound 8

[0282]

[0283] 11H NMR (400 MHz, CDCl3) δ8.19 (s, 1H), 8.04 (d, J=8.3 Hz, 1H), 7.81 (d, J=8.3 Hz, 1H), 7.61 (t, J=7.0 Hz, 1H), 7.51 (d, J=7.8 Hz, 1H), 7.40 (d, J=7.7 Hz, 1H), 7.33 (d, J=9.5 Hz, 1H), 6.70 (d, J=8.1 Hz, 1H), 5.51 (s, 2H), 5.21 (s, 1H), 4.78–4.59 (m, 3H), 4.36 (d, J=12.7 Hz, 3H), 4.15 (s, 2H), 3.88 (s, 2H), 3.28 (s, 2H), 2.85 (s, 2H), 2.77 (s, 2H), 2.73–2.67 (m, 1H), 2.45 (s, 1H).

[0284] Compounds 9-A and 9-B

[0285]

[0286] 1 1H NMR (400 MHz, MeOD) δ8.20 (s, 1H), 7.95 (dd, J=8.4, 1.3 Hz, 1H), 7.68–7.50 (m, 4H), 7.33 (d, J=8.4 Hz, 1H), 6.69 (d, J=8.4 Hz, 1H), 5.49 (dd, J=34.7, 13.8 Hz, 2H), 5.22 (tt, J=7.0, 3.5 Hz, 1H), 4.82–4.72 (m, 3H), 4.63 (ddd, J=15.7, 14.8, 5.5 Hz, 2H), 4.44 (dt, J=9.1, 6.0 Hz, 1H), 4.09 (d, J=13.7 Hz, 1H), 3.97 (d, J=13.7 Hz, 1H), 3.45 (td, J=9.9, 4.0 Hz, 1H), 3.07 (dd, J=10.0, 4.6 Hz, 2H), 2.74 (ddd, J=16.2, 8.8, 5.7 Hz, 1H), 2.58–2.44 (m, 3H), 2.35 (t, J=10.8 Hz, 1H), 2.20 (t, J=10.2 Hz, 1H), 1.37 (dd, J=12.3, 3.2 Hz, 1H).

[0287] 1¹H NMR (400 MHz, MeOD) δ 8.24 (s, 1H), 7.96 (dd, J=8.5, 1.4 Hz, 1H), 7.65 (dd, J=13.5, 8.1 Hz, 2H), 7.58–7.50 (m, 2H), 7.34 (d, J=8.4 Hz, 1H), 6.69 (d, J=8.4 Hz, 1H), 5.48 (dd, J=32.6, 13.7 Hz, 2H), 5.22 (dt, J=7.2, 4.3 Hz, 1H), 4.79 (d, J=3.6 Hz, 2H), 4.76–4.56 (m, 3H), 4.43 (dt, J=9.2, 6.0 Hz, 1H), 4.03 (q, J=13.7 Hz, 2H), 3.48 (td, J=10.0, 4.1 Hz, 1H), 3.16 (dd, J=9.9, 3.7 Hz, 1H), 2.97 (d, J=11.5 Hz, 1H), 2.80–2.71 (m, 1H), 2.50 (ddd, J=18.8, 12.2, 6.2 Hz, 3H), 2.28 (dt, J=20.2, 10.0 Hz, 2H), 1.40–1.30 (m, 1H).

[0288] Compound 10

[0289]

[0290] 1 ¹H NMR (400 MHz, CD₃OD_SPE) δ 8.17 (d, J=0.8 Hz, 1H), 7.95 (dd, J=8.4, 1.5 Hz, 1H), 7.59 (d, J=8.5 Hz, 2H), 7.47 (t, J=8.1 Hz, 1H), 7.26–7.10 (m, 2H), 6.69 (d, J=8.2 Hz, 1H), 5.42 (s, 2H), 5.26 (qd, J=7.1, 2.9 Hz, 1H), 4.88 (d, J=7.1 Hz, 1H), 4.70 (dd, J=15.3, 3.0 Hz, 1H), 4.63–4.56 (m, 1H), 4.45 (dt, J=9.1, 6.0 Hz, 1H), 4.34 (s, 2H), 4.10 (dd, J=37.2, 13.6 Hz, 2H), 3.86 (s, 2H), 3.23 (t, J=10.8 Hz, 2H), 2.85–2.70 (m, 5H), 2.57–2.47 (m, 1H).

[0291] Compound 11

[0292]

[0293] 1¹H NMR (400 MHz, MeOD) δ 8.17 (s, 1H), 7.95 (dd, J=8.4, 1.5 Hz, 1H), 7.69–7.52 (m, 5H), 6.75 (d, J=8.2 Hz, 1H), 5.54 (s, 2H), 5.26 (d, J=4.2 Hz, 2H), 4.70 (dd, J=15.4, 3.1 Hz, 1H), 4.61 (dd, J=13.7, 7.8 Hz, 1H), 4.45 (dt, J=9.1, 5.9 Hz, 1H), 4.35 (s, 2H), 4.10 (dd, J=34.7, 13.6 Hz, 2H), 3.87 (s, 2H), 3.48 (d, J=1.7 Hz, 1H), 3.24–3.23 (m, 1H), 2.85–2.71 (m, 5H), 2.53 (d, J=8.8 Hz, 1H).

[0294] Compounds 12A & 12-B

[0295]

[0296] ¹H NMR (400 MHz, MeOD) δ 8.17 (s, 1H), 7.93 (d, J=8.4 Hz, 1H), 7.68 (t, J=7.5 Hz, 1H), 7.61–7.52 (m, 3H), 7.41 (d, J=8.3 Hz, 1H), 6.63 (d, J=8.2 Hz, 1H), 5.56–5.44 (m, 2H), 5.24 (dt, J=6.9, 4.7 Hz, 1H), 5.00–4.90 (m, 1H), 4.74–4.60 (m, 4H), 4.52–4.32 (m, 2H), 4.13–3.86 (m, 2H), 3.76 (d, J=13.5 Hz, 1H), 3.21 (s, 1H), 2.96–2.75 (m, 2H), 2.74–2.26 (m, 6H), 1.65 (d, J=9.3 Hz, 1H).

[0297] ¹H NMR (400 MHz, MeOD) δ 8.16 (s, 1H), 7.94 (dd, J = 8.4, 1.2 Hz, 1H), 7.68 (t, J = 7.4 Hz, 1H), 7.62–7.50 (m, 3H), 7.41 (d, J = 8.3 Hz, 1H), 6.63 (d, J = 8.2 Hz, 1H), 5.55–5.42 (m, 2H), 5.26 (d, J = 7.0 Hz, 1H), 4.90 (s, 1H), 4.67 (ddd, J = 21.1, 16.6, 10.3 Hz, 4H), 4.44–4.25 (m, 2H), 4.02–3.78 (m, 3H), 3.21 (s, 1H), 2.93 (s, 1H), 2.85–2.73 (m, 1H), 2.62–2.27 (m, 6H), 1.66 (d, J = 9.3 Hz, 1H).

[0298] Compounds 13-A & 13-B

[0299]

[0300] 1 ¹H NMR (400 MHz, CD₃OD_SPE) δ 8.22 (s, 1H), 7.94 (d, J = 8.4 Hz, 1H), 7.61 (t, J = 8.4 Hz, 2H), 7.44–7.27 (m, 3H), 6.92–6.56 (m, 2H), 5.53–5.39 (m, 2H), 5.25 (d, J = 4.4 Hz, 1H), 4.56 (s, 5H), 4.42–4.28 (m, 2H), 3.98–3.83 (m, 3H), 3.26 (s, 1H), 2.94 (s, 1H), 2.78 (dt, J = 16.5, 8.1 Hz, 1H), 2.65–2.25 (m, 7H).

[0301] 1 ¹H NMR (400 MHz, MeOD) δ 8.16 (s, 1H), 7.93 (dd, J = 8.4, 1.2 Hz, 1H), 7.65–7.52 (m, 2H), 7.44–7.27 (m, 3H), 6.93–6.58 (m, 2H), 5.47 (q, J = 13.2 Hz, 2H), 5.25 (d, J = 4.9 Hz, 1H), 4.49 (ddd, J = 42.8, 42.1, 36.9 Hz, 7H), 4.08–3.89 (m, 2H), 3.76 (d, J = 13.5 Hz, 1H), 3.24 (s, 1H), 2.97–2.15 (m, 9H).

[0302] Compounds 14-A & 14-B

[0303]

[0304] 1 ¹H NMR (400 MHz, MeOD) δ 8.19 (s, 1H), 7.93 (d, J = 8.4 Hz, 1H), 7.68 (t, J = 7.6 Hz, 1H), 7.60–7.53 (m, 3H), 7.41 (d, J = 8.3 Hz, 1H), 6.63 (d, J = 8.2 Hz, 1H), 5.50 (s, 2H), 4.63 (s, 4H), 4.36 (d, J = 30.1 Hz, 2H), 4.23–4.14 (m, 1H), 4.12–4.02 (m, 1H), 3.95 (dd, J = 12.3, 4.4 Hz, 1H), 3.79 (dt, J = 22.5, 11.2 Hz, 2H), 3.21 (d, J = 8.9 Hz, 1H), 2.87 (s, 1H), 2.61 (s, 1H), 2.45–2.27 (m, 4H), 0.93–0.71 (m, 4H).

[0305] 1 ¹H NMR (400 MHz, MeOD) δ 8.19 (s, 1H), 7.93 (d, J = 8.4 Hz, 1H), 7.69 (t, J = 7.6 Hz, 1H), 7.60–7.52 (m, 3H), 7.41 (d, J = 8.2 Hz, 1H), 6.63 (d, J = 8.2 Hz, 1H), 5.56–5.42 (m, 2H), 4.62 (d, J = 12.4 Hz, 5H), 4.33 (s, 1H), 4.19 (d, J = 3.2 Hz, 1H), 4.06 (d, J = 3.2 Hz, 1H), 3.88 (ddd, J = 46.0, 22.4, 9.0 Hz, 3H), 3.21 (d, J = 9.4 Hz, 1H), 2.87 (s, 1H), 2.58 (d, J = 26.9 Hz, 1H), 2.45–2.27 (m, 4H), 0.98–0.67 (m, 4H).

[0306] Compound 15-A & 15-B

[0307]

[0308] 1 1H NMR (400 MHz, CD3OD_SPE) δ 8.15 (s, 1H), 7.93 (dd, J=8.4, 1.2 Hz, 1H), 7.70 (t, J=7.5 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.50–7.39 (m, 3H), 6.63 (d, J=8.2 Hz, 1H), 5.55–5.44 (m, 2H), 5.26 (dd, J=6.9, 2.7 Hz, 1H), 4.68 (ddd, J=29.4, 15.1, 7.2 Hz, 6H), 4.38 (ddd, J=28.7, 16.0, 8.4 Hz, 2H), 4.01–3.78 (m, 3H), 3.23 (d, J=10.1 Hz, 1H), 2.94 (s, 1H), 2.82–2.73 (m, 1H), 2.63–2.28 (m, 6H).

[0309]

[0309] 1 1H NMR (400 MHz, MeOD) δ 8.16 (s, 1H), 7.93 (d, J=8.5 Hz, 1H), 7.71 (t, J=7.3 Hz, 1H), 7.56 (d, J=8.4 Hz, 1H), 7.52–7.39 (m, 3H), 6.64 (d, J=8.2 Hz, 1H), 5.57–5.45 (m, 2H), 5.24 (d, J=5.3 Hz, 1H), 4.72–4.61 (m, 5H), 4.54–4.33 (m, 3H), 4.06–3.91 (m, 2H), 3.76 (d, J=13.5 Hz, 1H), 3.23 (s, 1H), 2.95–2.77 (m, 2H), 2.68 (s, 1H), 2.59–2.41 (m, 2H), 2.34 (d, J=8.1 Hz, 3H).

[0310] Compound 16

[0311]

[0312] 1H NMR (400MHz, CDCl3) δ8.19(q,J=8.2Hz,2H),7.61(t,J=7.5Hz,1H),7.52(d,J=8.3Hz,1H ),7.44(d,J=7.8Hz,1H),7.37(d,J=9.1Hz,1H),6.71(d,J=8.2Hz,1H),5.51(s,2H),5.25 (s,1H),4.93–4.85(m,1H),4.79(d,J=12.9Hz,1H),4.61(d,J=7.1Hz,1H),4.35(s,3H), 4.20(s,2H),3.87(s,2H),3.28(s,2H),2.84(d,J=5.4Hz,2H),2.74(s,3H),2.46(s,1H).

[0313] Compounds 17-A & 17-B

[0314]

[0315] Experiment 2 - In vitro activity test

[0316] (1) Testing instruments and reagents

[0317]

[0318]

[0319] (2) GLP-1R kit

[0320] GLP-1R-mediated agonist activity was determined using a homogeneous time-resolved fluorescence (HTRF) cAMP assay kit via a cell-based functional assay that measures cAMP levels in cells. This method is a competitive immunoassay. It enables the direct pharmacological characterization of compounds that act on Gs-coupled receptors in adherent or suspension cells.

[0321] The standard curve of naturally produced cAMP or unlabeled cAMP produced by cells competes with the red cAMP receptor labeled with d2 for binding to the monoclonal anti-cAMP cavitary compound europium donor. The specific signal is inversely proportional to the concentration of cAMP in the standard or experimental sample.

[0322] The human GLP-1R coding sequence (NCBI reference sequence NP_002053.3) was subcloned into pEGFP-N1 (tsingke), and cell lines stably expressing the receptor were isolated. The expression density of GLP-1R was confirmed by observing GFP expression under a fluorescence microscope.

[0323] (3) GLP-1R-GFP-293A cell culture

[0324] 293AGFP-GLP-1R cells were cultured in DMEM growth medium, 10% heat-inactivated fetal bovine serum (GEMINI Cat#900-108), and 1% Pen-3Trep (SangomBiotech Cat#E607011-0100) and in a humidified incubator at 37°C with 5% CO2.

[0325] (4) cAMP level testing method

[0326] Different concentrations of each analyte (in DMSO) were diluted 1:5 in distilled water with stimulation buffer. 500 μM 3-isobutyl-1-methylxanthin (IBMX; Meilunbiocat#MB5226) was added to obtain a 2X working solution. Then, 5 μL of the compound was added to a white 384-well assay plate (Corning 3824) using a multichannel pipette. The final DMSO concentration in the assay buffer mixture was 1‰.

[0327] Cells were collected from T25 tissue culture flasks and centrifuged at 1000 rpm for 5 minutes at room temperature. The cell pellet was then resuspended in 1 ml of stimulation buffer. A 20 μL sample of cell suspension was counted using a STARIC 1000 counter to determine cell viability and cell count per milliliter. The remaining cell suspension was then adjusted with stimulation buffer to deliver 2000 live cells per well using a multichannel pipette. 5 μL of cell suspension was added to each well of an assay plate already containing the compound. The plate was sealed and incubated at 37°C with 5% CO2 for 30 minutes.

[0328] After 30 minutes of incubation, 5 μL of d2-labeled cAMP and 5 μL of anti-cAMP cavitation compound (both diluted 1:20 in cell lysis buffer) were added to each well of the assay plate. The plate was then incubated at room temperature, and after 60 minutes, changes in the HTRF signal were read using a Tecan Spark plate reader, measuring absorbance at 340 nm excitation and 615 nm emission. The raw data were converted to nM cAMP by interpolation from the cAMP standard curve, and the percentage effect was determined relative to the saturation concentration of the full agonist GLP-17-37 (400 nM) contained on each plate. EC 50 The determination was performed from agonist dose-response curves, which were analyzed using a curve fitting procedure with a 4-parameter logical dose-response equation.

[0329] This experiment demonstrates that the compound of the present invention activates GLP-1R signaling via the cAMP pathway, thus acting as a GLP-1R agonist. Experimental data are presented as the geometric mean (EC50) based on the number of repetitions listed.50 The results are presented in the form of s).

[0330] (5) Experimental Results

[0331]

[0332]

[0333] Experiment 3 - Inhibition of hERG potassium ion channels

[0334] 1. Experimental materials: Stable cell line HEK-hERG, strain: HEK 293, source: Academy of Military Medical Sciences;

[0335]

[0336]

[0337] 2. Electrophysiological solutions

[0338] Extracellular fluid (mM): 10 N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES), 145 NaCl, 4 KCl, 2 CaCl2, 1 MgCl2, 1 Glucose, pH adjusted to 7.3–7.4 with sodium hydroxide; osmotic pressure adjusted to 290–310 mOsm; filtered and stored at 4°C.

[0339] Electrode internal solution (mM): KCl 120, KOH 31.25, CaCl2 5.374, MgCl2 1.75, ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) 10, HEPES 10, Na2-ATP 4, pH adjusted to 7.2-7.3 with potassium hydroxide; osmotic pressure adjusted to 290-310 mOsm; filtered and dispensed, stored at -20℃.

[0340] 3. Positive control compound:

[0341] Positive control: Amitriptyline hydrochloride or terfenadine

[0342] Source: Sigma-Aldrich

[0343] 4. Preparation of drug delivery formulations

[0344] Solvent control preparation: Add a certain volume of DMSO to the extracellular fluid to make it contain the same amount of DMSO as the final test solution (if the test solution contains different amounts of DMSO, use the highest DMSO content) to eliminate interference from DMSO on the cell's own electrical currents. Test sample preparation: Prepare a DMSO stock solution of the required concentration (generally 1000 / 3 times the actual drug concentration) by mixing the above 10mM stock solution with the specified ratio. Finally, dilute the stock solution with extracellular fluid to the required drug concentration for the experiment.

[0345] Preparation of positive control solution: Weigh an appropriate amount of positive control sample and place it in a suitable container. Add a certain volume of DMSO and stir or shake thoroughly to dissolve it completely. Prepare a 10mM stock solution. Then prepare the stock solution of the required concentration according to the ratio. Finally, dilute the stock solution with extracellular fluid to the required drug concentration for the experiment.

[0346] Before using the working concentration solution, check for any precipitation. If precipitation occurs, dilute the stock solution to increase the final DMSO concentration in the extracellular fluid, but the final DMSO concentration in the extracellular fluid should not exceed 0.5%. Continuous perfusion from low to high concentrations is used in the experiment. After the experiment, any remaining test sample and positive control solutions are disposed of as waste.

[0347] 5. Experimental Design

[0348] Cell preparation:

[0349] After passage and culture HEK-293-hERG cells to a suitable state, wash the cells with PBS (or DPBS), digest and separate them with Tryple solution, resuspend the cells in culture medium, and store them in centrifuge tubes. After centrifugation, discard the supernatant, resuspend the cells in extracellular fluid, and store at 2-8°C. Before patch-clamp recording, add cells to a culture dish to ensure a certain cell density and that the cells are individually separated.

[0350] Concentration settings:

[0351] Compounds of the present invention 1、10 Amitriptyline or Terfenadine 1

[0352] Electrophysiological experiments:

[0353] hERG currents were recorded using whole-cell patch-clamp technique. Cell suspension was added to a small culture dish and placed on the stage of an inverted microscope. After cell attachment, the cells were perfused with extracellular fluid at a recommended flow rate of 1–2 mL / min. The glass microelectrode was fabricated in two steps using a microelectrode drawing instrument; after filling with electrode fluid, its water resistance was 2–5 MΩ.

[0354] After establishing the whole-cell recording mode, the clamping potential was maintained at -80 mV. A depolarization voltage of +60 mV was applied for 850 ms, followed by repolarization to -50 mV for 1275 ms to extract the hERG tail current. This pulse program was repeated every 15 seconds throughout the experiment.

[0355] After the current stabilizes, a continuous extracellular perfusion administration method is adopted, starting from low concentrations and gradually increasing to high concentrations. Perfusion is continued from a low concentration until the efficacy stabilizes, and then the next concentration is applied. This experiment will test the blocking effect of each test sample and the positive control on the hERG tail current (N≥2); the specific actual concentration can be adjusted according to the actual solubility and effect, and this is not considered a deviation from the protocol.

[0356] The definition of stable efficacy is: if the change in current value of the last 5 stimulation strips in each concentration administration phase is less than 10% of the mean (when the current is greater than or equal to 200 pA) or less than 30% of the mean (when the current is less than 200 pA), it can be considered stable. If it is unstable, the concentration data will not be used.

[0357] 6. Data Analysis

[0358] In data processing, when determining the blocking effect on hERG, the peak value of the tail current and its baseline were corrected. The effect of each compound at different concentrations was expressed as the tail current inhibition rate (IR). A SD ≤ 15 for the %IR at all cell concentrations was considered acceptable (except for outlier data).

[0359] IR = 100% × (peak tail current before administration - peak tail current after administration) / peak tail current before administration.

[0360] 7. Experimental Results:

[0361]

[0362] 8. Experimental Conclusions

[0363] The compounds of this invention did not exhibit hERG inhibitory activity.

[0364] Experiment 4 - Liver microsomal metabolic stability

[0365] 1. Experimental Design: Concentration to be measured: 1 μM; Control compound: Testosterone; Culture conditions: Incubated at 37℃ for 0, 5, 15, 30, and 45 minutes; Assay method: LC-MS / MS; Calculation method: T 1 / 2 = 0.693 / K (K is the rate constant of ln[concentration] versus incubation time), Cl int =(0.693 / T) 1 / 2)×(1 / (microsomal protein concentration (0.5mg / mL)))×proportional factor.

[0366] 2. Experimental Methods: 1. Preheat 0.1M K-buffer, 5nM MgCl2, pH=7.4; 2. Experimental solutions for test and reference compounds: 500μM additive solution: Add 5μL of 10 mM stock solution to 95μL ACN; 1.5μM microparticle doping solution (0.75mg / mL): Add 1.5μL of 500μM additive solution and 18.75μL of 20Mg / mL liver microparticles to 479.75μL K / Mg buffer; 3. 3×NADPH stock solution (6mM, 5mg / mL) is NADPH dissolved in buffer; 4. Dispense 30μL of 1.5μM additive solution containing 0.75mg / mL microparticle solution onto the assay plate designated for different time points (0, 5, 15, 30, 45 minutes); 5. At 0 minutes, add 150μL of ACN containing IS. 6. Add 15 μL of NADPH stock solution (6 mM, step 3) to the wells of the plate; 7. Pre-incubate all other plates at 37°C for 5 minutes; 8. Add 15 μL of NADPH stock solution to the plate to start the reaction and start timing; 9. At 5 minutes, 15 minutes, 30 minutes, and 45 minutes, add 150 μL of ACN containing IS to the wells of the corresponding plates to terminate the reaction; 10. After quenching, shake the plate on a shaker for 10 minutes (600 rpm / min), and then centrifuge at 6000 rpm for 15 minutes; 11. Transfer 80 μL of supernatant from each well to a 96-well sample plate containing 140 μL of water for LC / MS analysis.

[0367] 3. Analysis Methods:

[0368] Detection method: LC-MS / MS-11(8050), internal standard: tolbutamide, MS conditions: testosterone and analyte positive ion ESI; tolbutamide negative ion ESI; mobile phase: mobile phase A is 0.1% FA in water, mobile phase B is 0.1% FA in ACN; column and specifications: ACQUITY UPLC HSS T31.8um 2.1*50mm.

[0369] LC conditions:

[0370]

[0371] 4. Experimental Results (Human Microsomes):

[0372] 2 41.82 2P-1 148.12 2P-2 23.18 3P-1 16.21 4P-1 37.81 5P-1 94.3 6 68.21 7 180.00 8 82.73 9 399.45 9-B 338.86 10 25.70 11 39.85 12 100.74 16 56.64

[0373] 5. Experimental Conclusion: The compound of this invention exhibits good stability in liver microsomes.

[0374] Experiment 5 - Caco-2 cell transport experiment

[0375] 1. Experimental materials:

[0376] Caco-2 cells, passage 77; HBSS, Lot: G210713; ACN+IS (tolbutamide 200 ng / ml);

[0377] 2. Cell culture:

[0378] Caco-2 cells were seeded at 2 × 10⁵ cells / cm² onto polyethylene (PET) membranes in 96-well Falcon plates until a confluent cell monolayer formed after 21–28 days. The culture medium was changed every 3–4 days.

[0379] 3. Experimental Design:

[0380] The test compound was diluted to 10 μM with 10 mM stock solution in transport buffer (HBSS without BSA) and applied to the apical or basal side of a cell monolayer. Incubation was performed at 37°C and 5% CO2 for 120 min at 95% relative humidity. The permeation of the test compound from A to B or B to A was measured in duplicate. The elution ratio of each compound was also determined. Based on the analyte / IS peak area ratio, the test and reference compounds were quantified by LC-MS / MS analysis.

[0381] 4. Experimental measurements:

[0382] The apparent permeability coefficient Papp (cm / s) is calculated using the following equation:

[0383] Papp = (dCr / dt) x Vr / (A × C0), where dCr / dt is the cumulative concentration of the compound in the acceptor chamber, which is a function of time (s), Vr is the volume of the solution in the acceptor chamber (0.1 mL on the top side and 0.25 mL on the bottom side), A is the surface area used for transport, i.e., 0.0804 cm2 is the area of ​​the monolayer, and C0 is the initial concentration in the donor chamber;

[0384] The outflow ratio is calculated using the following formula:

[0385] EffluxRatio=Papp(BA) / Papp(AB);

[0386] The recovery percentage is calculated using the following equation:

[0387] %Recovery=100×[(Vr×Cr)+(Vd×Cd)] / (Vd×C0)

[0388] %Total recovery=100×[(Vr×Cr)+(Vd×Cd)+(Vc×Cc)] / (Vd×C0),

[0389] Where Vd is the volume in the donor chamber (0.1 mL on the top side and 0.25 mL on the bottom side), Cd and Cr are the final concentrations of the transported compounds in the donor and recipient chambers, respectively, Cc is the concentration of the compound in the cell lysate solution, and Vc is the volume of the insert well (0.1 mL in this experiment).

[0390] 5. LC / MS conditions:

[0391] Detection methods: LC-MS / MS-20 (TQ-6500+) & LC-MS / MS-11 (8050), internal standard: tolbutamide, MS conditions: atenolol and propranolol and the cation ESI of the compounds, digoxin anion ESI; mobile phase: mobile phase A is 0.1% FA in water, mobile phase B is 0.1% FA in ACN; column and specifications: ACQUITYUPLC HSS T31.8um 2.1*50mm.

[0392] LC conditions:

[0393]

[0394]

[0395] 6. Experimental Results:

[0396] 4-P1 1.54 / 9.36 / 6.08 6 0.98 / 11.05 / 11.27

[0397] 7. Experimental Conclusion:

[0398] The compounds of this invention are well absorbed in the intestine.

[0399] Preparation Examples

[0400] The intermediate reactants used in the preparation process were prepared according to the preparation method described in WO2018109607A1.

[0401] The preparation method of intermediate 1-int-2,(S)-2-(chloromethyl)-1-(oxetanebut-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid methyl ester is as follows:

[0402]

[0403] (1) Preparation of compound 1-2C

[0404]

[0405] Me3SO4 was added in portions to a stirred solution of t-BuOK (170 g, 1520 mmol, 2.5 equivalents) in t-BuOH (500 mL) at 60 °C under an argon atmosphere. + I - (335 g, 1520 mmol, 2.5 equivalents), after 30 minutes, (S)-2-((benzyloxy)methyl)ethylene oxide 1-1C (100 g, 610 mmol, 1.00 equivalents) was added dropwise to the above mixture. The resulting mixture was stirred at 60 °C for another 13 hours. The mixture was cooled to room temperature and then filtered, and the filter cake was washed with EtOAc (3 × 200 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4 and concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography, eluting with PE / EtOAc (10:1) to give (S)-2-((benzyloxy)methyl)oxetine, 1-2C (50.0 g, 46% yield).

[0406] 1 H NMR (400MHz, CDCl3) δ = 7.39–7.26 (m, 5H), 5.04–4.90 (m, 1H), 4.73–4.50 (m, 4H), 3.64 (qd, J = 11.0, 4.3Hz, 2H), 2.72–2.45 (m, 2H).

[0407] (2) Preparation of compound 1-3C

[0408]

[0409] A solution of (S)-2-((benzyloxy)methyl)oxetane 1-2C (50 g, 280.9 mmol, 1.0 equivalent) and Pd / C (20 g, wet) in THF (200 mL) was stirred at 50 °C for 16 hours under H2 (4 MPa). The mixture was cooled to room temperature and then filtered, with the filter cake washed with THF (100 mL). The filtrate was concentrated under reduced pressure to give (S)-oxetane-2-ylmethanol, 1-3C (28 g, crude product), which was used directly in the next step.

[0410] (3) Preparation of compounds 1-4C

[0411]

[0412] At 25 °C, TsCl (66.6 g, 349.6 mmol, 1.1 eq) and TEA (48.2 g, 476.7 mmol, 1.5 eq) were added to a THF (200 mL) solution of (S)-oxetane-2-ylmethanol 1-3C (28 g, 317.8 mmol, 1 eq). The mixture was stirred at room temperature for 2 hours. The mixture was diluted with H2O (100 mL) and extracted with DCM (100 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated to give a residue, which was purified by column chromatography on silica gel, eluting with (EA / PE = 0-10%) to give (S)-oxetane-2-ylmethyl 4-methylbenzenesulfonate, 1-4C (56 g, 72.7% yield), as a colorless oil.

[0413] 1 H NMR(400MHz, CDCl3)δ=7.85–7.79(m,2H),7.35(dd,J=8.6,0.6Hz,2H),5.00–4.83(m,1H),4.6 8–4.38(m,2H),4.16(d,J=4.0Hz,2H),2.78–2.64(m,1H),2.58(d,J=9.0Hz,1H),2.45(s,3H).

[0414] (4) Preparation of compounds 1-5C

[0415]

[0416] To a DMF (200 mL) solution of (S)-oxetane-2-ylmethyl-4-methylbenzenesulfonate 1-4C (56 g, 231 mmol, 1 eq), NaN3 (22.5 g, 346.7 mmol, 1.5 eq) was added. The mixture was stirred at 60 °C for 12 hours. The mixture was diluted with H2O (100 mL) and extracted with EtOAc (100 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated to give (S)-2-(azidomethyl)oxetane, 1-5C (20 g, crude product), which was used directly in the next step.

[0417] (5) Preparation of compounds 1-6C

[0418]

[0419] A solution of (S)-2-(azidomethyl)oxetane 1-5C (20 g, crude product) and Pd / C (8 g) in THF (100 mL) was stirred at 25 °C for 16 hours under H2 (15 Psi). The resulting mixture was filtered, and the filter cake was washed with THF (3 × 100 mL). The filtrate was directly concentrated to give (S)-oxetane-2-ylmethylamine, 1-6C (3.8 g, crude product).

[0420] 1 H NMR (400MHz, DMSO) δ = 4.60 (dq, J = 6.5, 5.2Hz, 1H), 4.52–4.43 (m, 1H), 4.40–4.30 (m ,1H),2.67(t,J=5.5Hz,2H),2.57–2.51(m,1H),2.38(ddt,J=10.8,9.0,7.0Hz,2H).

[0421] (6) Preparation of compounds 1-7C

[0422]

[0423] At 25 °C, methyl 3-fluoro-4-nitrobenzoate 1-6D (8.69 g, 43.6 mmol, 1.0 eq) and TEA (8.83 g, 87.2 mmol, 2 eq) were added to a THF (80 mL) solution of (S)-oxetane-2-ylmethylamine 1-6C (3.8 g, 43.6 mmol, 1 eq). The mixture was stirred at 40 °C for 6 hours. The mixture was concentrated to obtain a residue, which was purified by silica gel column chromatography by elution with (EtOAc / petroleum ether = 0-80%) to give methyl (S)-4-nitro-3-((oxetane-2-ylmethyl)amino)benzoate, 1-7C (6.2 g, 53.4% ​​yield).

[0424] 1 H NMR (400MHz, CDCl3) δ = 8.36 (s, 1H), 8.23 ​​(d, J = 8.9Hz, 1H), 7.63 (d, J = 1.4Hz, 1H), 7.26 (dd, J = 8.8, 1.7Hz, 1H), 5.1 6(tt,J=7.4,4.5Hz,1H),4.81–4.55(m,2H),3.94(s,3H),3.71–3.55(m,2H),2.84–2.72(m,1H),2.70–2.52(m,1H).

[0425] (7) Preparation of compounds 1-8C

[0426]

[0427] A solution of methyl (S)-4-nitro-3-((oxetane-2-ylmethyl)amino)benzoate 1-7C (6.2 g, 23.3 mmol, 1.0 eq) and Pd / C (1.0 g, wet) in MeOH (100 mL) was stirred at 25 °C under H₂ (1 atm) for 12 h. The mixture was filtered, and the filter cake was washed with MeOH (3 × 20 mL). The filtrate was directly concentrated to give methyl (S)-4-amino-3-((oxetane-2-ylmethyl)amino)benzoate 1-8C (5.2 g, 94.5% yield). LCMS: rt = 1.201 min, [M+1] + =237.1, purity: 89.7%.

[0428] (8) Preparation of compound 1-int-2

[0429]

[0430] Add 2-chloro-1,1,1-trimethoxyethane 1-8D (0.98 g, 6.35 mmol, 1.5 eq) and TsOH·H₂O (0.08 g, 0.423 mmol, 0.1 eq) to a THF (20 mL) solution of (S)-4-amino-3-((oxetane-2-ylmethyl)amino)benzoate 1-8C (1.0 g, 4.23 mmol, 1 eq). Stir the mixture at 50 °C for 8 hours. Dilute the mixture with saturated sodium bicarbonate solution. Extract with NaHCO₃ (20 mL) and EtOAc (10 mL × 3). The combined organic layers were dried over Na2SO4, filtered and concentrated to obtain the residue, which was purified by silica gel column chromatography and eluted with (EtOAc / petroleum ether = 0-80%) to give (S)-2-(chloromethyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylate, methyl 1-int-2 (1.1 g, 88% yield).

[0431] 1 H NMR (400MHz, CDCl3) δ8.12(d,J=0.9Hz,1H),8.01(dd,J=8.5,1.5Hz,1H),7.79(d,J=8.5Hz,1H),5.21(ddd,J=9.6,7.3,2.7Hz,1H),5.03 (s,2H),4.69–4.45(m,3H),4.34(d,J=9.2Hz,1H),3.96(s,3H),2.76(dtd,J=11.5,8.1,6.0Hz,1H),2.42(ddt,J=11.5,9.2,7.3Hz,1H).

[0432] The preparation method of intermediate methyl 1-int-3,2-(chloromethyl)-1-((1-(fluoromethyl)cyclopropyl)methyl)-1H-benzo[d]imidazolium-6-carboxylate is as follows:

[0433]

[0434] (1) Preparation of 3a-1

[0435]

[0436] Under nitrogen atmosphere and at 80 °C, a solution of methyl 3-fluoro-4-nitrobenzoate (2.0 g, 10.05 mmol) in DMF (30 mL) was mixed with (1-(aminomethyl)cyclopropyl)methanol (1.015 g, 10.05 mmol) and K₂CO₃ (2.087 g, 15.075 mmol) for 16 hours, and the reaction was monitored by LCMS. The reaction mixture was poured into water (300 mL), extracted with EtOAc (80 mL × 3), washed with brine and dried, and concentrated to give crude product. This crude product was further purified by elution (PE / EtOAc = 0-30%) to give 3a-1 (2.5 g, 89.2% yield).

[0437] LCMS: rt = 1.3 min, [M+1] + =281, Purity: 92%

[0438] (2) Preparation of 3a-2

[0439]

[0440] Under nitrogen atmosphere and at 25°C, Pd / C (1.25 g) was added to 100 mL of MeOH solution containing 3a-1 (2.5 g, 8.6 mmol), and the mixture was stirred for 2 hours. The reaction was monitored by LC-MS. The reaction mixture was filtered, and the filtrate was concentrated to give crude product 3a-2 (1.7 g, 76.2% yield). The crude product was used directly in the next step without purification.

[0441] LCMS: rt = 0.8 min, [M+1] + =251, Purity: 92%

[0442] (3) Preparation of 3a-3

[0443]

[0444] At 50 °C, 2-chloro-1,1,1-trimethoxyethane (3.142 g, 20.4 mmol) and TsOH (129.2 mg, 0.68 mmol) were added to 30 mL of THF solution of 3a-2 (1.7 g, 6.8 mmol) with stirring and mixed for 16 hours. The reaction was monitored by LCMS. The reaction mixture was quenched by adding saturated NaHCO3 aqueous solution (50 mL), extracted with EtOAc (30 mL × 3), washed with brine and dried, concentrated to give crude product, which was further purified by elution (EA / PE = 10–51%) to give 3a-3 (1.5 g, 71% yield).

[0445] LCMS: rt = 1.5 min, [M+1] + =309, Purity: 90%

[0446] (4) Preparation of 1-int-3

[0447]

[0448] Under a nitrogen atmosphere, the mixture was stirred at -65°C for 3 hours, and then added to a solution of 3a-3 (500 mg, 1.62 mmol) in 15 mL of DAST / THF. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by pre-HPLC to give 1-int-3 (400 mg, 79.52% yield). LCMS: rt = 1.708 min, [M+1] + =311, Purity: 90%

[0449] The chiral pre-SFC method is as follows:

[0450] System: Waters SFC 150

[0451] Column: Dr.maish Reprosil Chiral-MIC( IC)

[0452] Column size: 250*25mm, 10m

[0453] Mobile phase A is supercritical CO2, and mobile phase B is MEOH (±0.1% 7.0 mol / L ammonia solution in MEOH), with a ratio of A:B = 50:50.

[0454] Wavelength: 214nm

[0455] Flow rate: 120 ml / min

[0456] Column temperature: normal temperature

[0457] Back pressure: 100 bar

[0458] Injection volume: 4mL

[0459] Cycle time: 10 min

[0460] Sample preparation method: Dissolve the sample in approximately 20 mL of MeOH.

[0461] Example 1

[0462] 2-((2-((4-cyano-2-fluorobenzyl)oxy)-5-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazin-8-yl)methyl)-1-(((S)-oxetanebut-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (Compound 1)

[0463]

[0464] (1) Preparation of compounds 1-2

[0465]

[0466] At 25 °C, 1-(tert-butyl)-3-methylpiperazine-1,3-dicarboxylic acid ester and (i.e., 1A) (1.3 g, 5.2 mmol, 1 eq) and TEA (0.786 g, 7.77 mmol, 1.5 eq) were added to a THF (20 mL) solution of 2,6-dichloro-3-nitropyridin-2-yl)piperazine-1,3-dicarboxylic acid ester, compound 1-2 (1.7 g, 81.7% yield). The mixture was stirred at room temperature for 2 hours. The mixture was concentrated and the residue was purified by column chromatography on silica gel, eluting with (PE / EA = 0-50%) to give 1-(tert-butyl)-3-methyl-4-(6-chloro-3-nitropyridin-2-yl)piperazine-1,3-dicarboxylic acid ester.

[0467] LCMS: rt = 2.953 min, [M-55]+ = 345.0, purity: 90%.

[0468] (2) Preparation of compounds 1-3

[0469]

[0470] At 25°C, Zn (2.6 g, 40 mmol, 10 eq) was added to a solution of 1-(tert-butyl)3-methyl 4-(6-chloro-3-nitropyridin-2-yl)piperazine-1,3-dicarboxylic acid (i.e., 1-2) (1.6 g, 4 mmol, 1 eq) in HOAc (20 mL). The mixture was stirred at 60°C for 2 hours. The mixture was diluted with NH3·H2O (30 mL) and extracted with EtOAc (30 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and concentrated under reduced pressure to obtain the residue, which was purified by silica gel column chromatography by elution with PE / EtOAc (1:1) to give tert-butyl 2-chloro-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazino-8-carboxylic acid as compounds 1-3 (0.8 g, 59.2% yield).

[0471] LCMS: rt = 1.549 min, [M-55]+ = 283.0, purity: 81.3%.

[0472] (3) Preparation of compounds 1-4

[0473]

[0474] At 0 °C, NaH (60%, 266.6 mg, 6.66 mmol) was added to a 20 mL LDM solution of 2-chloro-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazino-8-carboxylic acid tert-butyl ester (i.e., 1-3) (1.5 g, 4.44 mmol), and the mixture was stirred at room temperature under nitrogen for 15 min. Then, CH3I (755.7 mg, 5.33 mmol) was added to the reaction mixture at room temperature over 30 min. The reaction mixture was poured into water (50 mL) and extracted with EtOAc (80 mL × 3). The organic layers were combined, washed with brine (20 mL), dried over Na2SO4, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography. Elution with (PE / EA = 0-30%) yielded tert-butyl 2-chloro-5-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazino-8-carboxylate, compounds 1-4 (1.44 g, 92.2% yield).

[0475] LCMS: rt = 2.083 min, [M-55]+ = 297.2, purity 90.8%.

[0476] (4) Preparation of compounds 1-5

[0477]

[0478] Under a nitrogen atmosphere and at 110 °C, tert-butyl 2-chloro-5-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazino-8-carboxylate (i.e., 1-4) (1.44 g, 4.1 mmol) was mixed with KOH (688.8 mg, 12.3 mmol), t-BuXphos (348 mg, 0.82 mmol), and Pd2(dba)3 (187 mg, 0.2 mmol) in 10 mL of dioxane / H2O (5:1) and stirred for 16 hours. The mixture was adjusted to pH 5 with HCl (1N) and extracted with EtOAc (80 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and concentrated under reduced pressure to obtain the residue, which was purified by silica gel column chromatography by elution with PE / EtOAc (1:1) to give tert-butyl 2-hydroxy-5-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazino-8-carboxylate, compound 1-5 (0.73 g, 53% yield).

[0479] LCMS: rt = 1.643 min, [M-55]+ = 279.0, purity: 82.7%.

[0480] (5) Preparation of compounds 1-6

[0481]

[0482] At 0 °C, NaH (60%, 0.043 g, 1 mmol, 1.2 eq) was added to a DMF (10 mL) solution of 2-hydroxy-5-methyl-6-oxo-5,6,6A,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazino-8-carboxylic acid tert-butyl ester (i.e., 1-5) (0.3 g, 0.9 mmol, 1 eq), followed by the addition of 4-(bromomethyl)-3-fluorobenzyl nitrile (i.e., 5A) (0.19 g, 0.9 mmol, 1 eq). The mixture was stirred at room temperature for 2 hours. The mixture was diluted with H2O (10 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated to obtain a residue, which was purified by silica gel column chromatography by elution with (PE / EA = 0-80%) to give 2-((4-cyano-2-fluorobenzyl)oxy)-5-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazine-8-carboxylate, compounds 1-6 (0.25 g, 59.6% yield).

[0483] LCMS: rt = 1.304 min, [M-55]+ = 412.1, purity: 88.6%.

[0484] (6) Preparation of compounds 1-7

[0485]

[0486] A solution of tert-butyl 2-((4-cyano-2-fluorobenzyl)oxy)-5-methyl-6-oxo-5,6,6A,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazin-8-carboxylate (i.e., 1-6) (0.25 g, 0.53 mmol, 1 eq) in HCl / EtOAc (10 mL) was stirred at room temperature for 2 hours. The mixture was concentrated to give 3-fluoro-4-(((5-methyl-6-oxo-6,6a,7,8,9,10-hexahydro-5H-pyrazino[1,2-a]pyridino[3,2-e]pyrazin-2-yl)oxy)methyl)benzyl nitrile hydrochloride, compounds 1-7 (0.15 g, 76.5% yield).

[0487] LCMS: rt = 1.128 min, [M+1]+ = 368.1, purity: 54.5%.

[0488] (7) Preparation of compounds 1-8

[0489]

[0490] To a solution of 3-fluoro-4-(((5-methyl-6-oxo-6,6a,7,8,9,10-hexahydro-5H-pyrazino[1,2-a]pyridino[3,2-e]pyrazino-2-yl)oxy)methyl)benzyl nitrile hydrochloride (i.e., 1-7) (0.15 g, 0.41 mmol, 1 eq) in CH3CN (10 mL), methyl (S)-2-(chloromethyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylate (i.e., 1-int-2) (0.12 g, 0.41 mmol, 1 eq) and DIEA (0.26 g, 2.04 mmol, 5.0 eq) were added. The mixture was stirred at 60 °C for 12 hours. The mixture was purified by silica gel column chromatography and eluted with (PE / EA = 0-50%) to give methyl 2-((2-(((4-cyano-2-fluorobenzyl)oxy)-5-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazin-8-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate, compounds 1-8 (0.2 g, 78.4% yield).

[0491] LCMS: rt=1.808min, [M+1]+=626.3, purity: 93.9%.

[0492] (8) Preparation of Compound 1

[0493]

[0494] At 25 °C, a solution of LiOH (0.035 g, 1.44 mmol, 5 eq) in H₂O (3 mL) was added to a solution of methyl 2-((2-((4-cyano-2-fluorobenzyl)oxy)-5-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazin-8-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (i.e., 1-8) (0.18 g, 0.29 mmol, 1 eq) in THF (3 mL). The mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated to obtain a residue, which was purified by pre-HPLC under alkaline conditions to give 2-((2-((4-cyano-2-fluorobenzyl)oxy)-5-methyl-6-oxo-5,6,6a,7,9,10-hexahydro-8H-pyrazino[1,2-a]pyridino[3,2-e]pyrazin-8-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid, compound 1 (30.8 mg, 16.02% yield).

[0495] LCMS: rt = 1.578 min, MH+ = 612.3, purity: 100%.

[0496] 1 H NMR (400MHz, CD3OD) δ = 8.32 (s, 1H), 7.97 (d, J = 7.6Hz, 1H), 7.75–7.61 (m, 2H), 7.59–7.48 (m, 2H), 7.29 (d d,J=8.4,1.7Hz,1H),6.28(dd,J=8.4,2.0Hz,1H),5.54–5.36(m,2H),5.28–5.17(m,1H),4.71–4.61(m,3 H),4.44(dt,J=9.1,5.9Hz,1H),4.33–4.21(m,1H),4.07(d,J=13.7Hz,1H),4.02–3.80(m,2H),3.37(d,J =11.3Hz,1H),3.25(d,J=6.3Hz,3H),3.04–2.69(m,3H),2.51(dd,J=17.8,9.7Hz,1H),2.38–2.13(m,2H).

[0497] Example 2

[0498] 2-((2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (compound 2)

[0499]

[0500] (1) Preparation of compound 2-1

[0501]

[0502] Cesium carbonate (6.06 g, 18.6 mmol), Xantphos (1.04 g, 1.8 mmol), and Pd2(dba)3 (820 mg, 0.9 mmol) were added to a mixture of 3-(hydroxymethyl)piperazine-1-carboxylic acid tert-butyl ester SM (2 g, 9.3 mmol) and 2,6-dichloro-3-fluoropyridine (1.53 g, 9.3 mmol) in 1,4-dioxane (20 mL) under a nitrogen atmosphere. The mixture was stirred at 100 °C for 7 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The reaction mixture was quenched with water and extracted with EA. The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography and eluted with PE / EA (1:1) to give tert-butyl 4-(6-chloro-3-fluoropyridin-2-yl)-3-(hydroxymethyl)piperazine-1-carboxylate, compound 2-1 (2.2 g, 60% yield).

[0503] LCMS:[M+1] + =346; Purity: 91%.

[0504] (2) Preparation of compound 2-2

[0505]

[0506] At 25 °C, potassium tert-butoxide (545 mg, 4.86 mmol) was added to a DMSO (5 mL) solution of 1.4 g (4.05 mmol) of 4-(6-chloro-3-fluoropyridin-2-yl)-3-(hydroxymethyl)piperazin-1-carboxylate (i.e., 2-1). The mixture was stirred for 5 h under nitrogen atmosphere and at 70 °C. The reaction was monitored by LCMS. The mixture was diluted with MeOH (5 mL), and the crude product was purified by pre-HPLC under alkaline conditions (ACN / water = 30-80%) to give tert-butyl 2-chloro-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-8(6H)-carboxylate, compound 2-2 (450 mg, 35% yield).

[0507] LCMS:[M+1] + =326; Purity: 97%.

[0508] 1H-NMR (400MHz, CDCl3) δ6.90(d,J=8.0Hz,1H),6.57(d,J=8.0Hz,1H),4.48(d,J=12.3Hz,1H),4.31–4.00( m,3H),3.92(dd,J=11.0,8.2Hz,1H),3.35(ddt,J=11.3,8.1,3.2Hz,1H),3.05–2.50(m,3H),1.48(s,9H).

[0509] (3) Preparation of compounds 2-3

[0510]

[0511] At 25 °C, KOH (155 mg, 2.76 mmol), tBuXPhos (119 mg, 0.28 mmol), and Pd2(dba)3 (128 mg, 0.14 mmol) were added to a mixture of 2-chloro-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazine-8(6H)-carboxylic acid tert-butyl ester (i.e., 2-2) (450 mg, 1.38 mmol) in 1,4-dioxane (10 mL). The mixture was stirred at 110 °C for 8 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The reaction mixture was quenched with water, the mixture was extracted with EA, the combined organic layers were washed with brine (30 mL), dried over Na2SO4 and concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography by elution with DCM / MeOH (3:1) to give tert-butyl 2-hydroxy-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazine-8(6H)-carboxylate, compounds 2-3 (200 mg, 40% yield).

[0512] LCMS:[M+1] + =308; Purity: 76%.

[0513] (4) Preparation of compounds 2-4

[0514]

[0515] Sodium hydride (19 mg, 0.46 mmol) was added to a DMF (3 mL) solution of 2-hydroxy-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazine-8(6H)-carboxylic acid tert-butyl ester (i.e., 2-3) (70 mg, 0.23 mmol). The mixture was stirred for 15 min under a nitrogen atmosphere, and then 4-(bromomethyl)-3-fluorobenzyl nitrile (49 mg, 0.23 mmol) was added. The mixture was stirred at room temperature for 3 h under a nitrogen atmosphere. The reaction was monitored by LCMS. The reaction mixture was quenched with ice water, extracted with EA, and the combined organic layers were washed with brine (30 mL), dried over Na2SO4 and concentrated under reduced pressure to obtain the residue, which was purified by silica gel column chromatography by elution with PE / EA (20:1) to give tert-butyl 2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azine-8(6H)-carboxylate, compound 2-4 (76 mg, 75% yield).

[0516] LCMS:[M+1] + =441; Purity: 83%.

[0517] (5) Preparation of compounds 2-5

[0518]

[0519] A solution of 2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azin-8(6H)-carboxylic acid tert-butyl ester (i.e., 2-4) (76 mg, 0.04 mmol) and HCl in EtOAc (4 M, 1 mL) was stirred at 25 °C under nitrogen for 3 h. The reaction was monitored by LCMS. The reaction mixture was concentrated under reduced pressure to give 3-fluoro-4-(((6,6a,7,8,9,10-hexahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-2-yl)oxy)methyl)benzyl nitrile (60 mg, crude product, 102% yield), the hydrochloride salt of compounds 2-5.

[0520] LCMS:[M+1] + =341; Purity: 62%.

[0521] (6) Preparation of compounds 2-6

[0522]

[0523] At 25 °C, methyl (S)-2-(chloromethyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylate (i.e., 1-int-2) (35 mg, 0.12 mmol) was added to a mixture of 3-fluoro-4-(((6,6a,7,8,9,10-hexahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-2-yl)oxy)methyl)benzyl nitrile (i.e., 2-5) (50 mg, 0.15 mmol) and DIEA (97 mg, 0.75 mmol) in ACN (5 mL). The mixture was stirred at 60 °C for 3 h under a nitrogen atmosphere. The reaction was monitored by LCMS. The reaction mixture was quenched with water and extracted with EA. The combined organic layers were washed with brine (30 mL), dried over Na2SO4 and concentrated under reduced pressure to give the residue, which was purified by silica gel column chromatography by elution with PE / EA (1:5) to give 2-((2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyrido[3,2-b][1,4]azin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid ester, compound 2-6 (30 mg, 42% yield).

[0524] LCMS:[M+1] + =599; Purity: 75%.

[0525] (7) Preparation of compound 2

[0526]

[0527] At 25 °C, lithium hydroxide (7 mg, 0.25 mmol) was added to a mixture of methyl 2-((2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (i.e., 2-6) (30 mg, 0.05 mmol) in THF (5 mL) and H2O (1 mL). The mixture was stirred at 25 °C for 3 hours. The reaction was monitored by LCMS. The mixture was purified by pre-HPLC (0.05% NH3·H2O / ACN: 20%-35%) to give 2-((2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyrido[3,2-b][1,4]azin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid, compound 2 (7.7 mg, 28% yield).

[0528] LCMS:[M+1] + =585; Purity: 96.21%

[0529] 1 H-NMR (400MHz, MeOD) δ8.17(s,1H),7.94(d,J=8.5Hz,1H),7.57(dddd,J=8.9,8.0,6.6,5.3Hz,4H),6.94( dd,J=8.3,0.7Hz,1H),6.07(dd,J=8.3,1.7Hz,1H),5.38(s,2H),5.27(dd,J=7.0,3.3Hz,1H),4.81(d,J=6 .6Hz,1H),4.75–4.58(m,2H),4.51–4.40(m,1H),4.37–4.28(m,1H),4.19–3.77(m,4H),3.34(d,J=11.7Hz ,1H),3.03–2.69(m,4H),2.52(dd,J=17.0,10.1Hz,1H),2.32–2.20(m,1H),2.00(dt,J=16.1,10.8Hz,1H).

[0530] Preparation methods of compounds 2-6-P1 and 2-6-P2

[0531]

[0532] Methyl 2-((2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azine-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate 2-6 (220 mg, 0.37 mmol) was administered via SFC. One-step purification yielded methyl 2-(((R)-2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azine-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate 2-6-P1 (90 mg, SFC) rt = 2.47 min, yield 41%) and methyl 2-(((S-2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyrido[3,2-b][1,4]azine-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate 2-6-P2 (80 mg, SFC rt = 3.34 min, yield 36%).

[0533] The SFC method is as follows:

[0534] Sample name: MT89_02-MC20-1147-030; System: Waters SFC 150; Column: AD; Column diameter: 250*25mm 10m;

[0535] Mobile phase A: supercritical CO2; Mobile phase B: MeOH (±0.1% ±7.0 mol / L ammonia in MeOH); A:B: 50:50; Wavelength: 214 nm; Flow rate: 100 mL / min; Column temperature: room temperature; Back pressure: 100 bar; Injection volume: 1 mL; Circulation time: 5 min;

[0536] Preparation of sample solution: Dissolve the sample in approximately 20 mL of MeOH / DCM (2:1).

[0537] Preparation methods of compounds 2-P1 and 2-P2

[0538]

[0539] At 25°C, lithium hydroxide (18 mg, 0.75 mmol) was added to a mixture of methyl 2-(((R)-2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azine-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate 2-6-P1 (90 mg, 0.15 mmol) in THF (5 mL) and H2O (1 mL). The mixture was stirred at 25°C for 20 hours. The reaction was detected by LCMS. The mixture was pre-purified by HPLC (0.05% NH3·H2O / ACN: 20%-35%) to give 2-(((R)-2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid, compound 2-P1 (43 mg, 50%), which was a grayish-white solid and was confirmed by pure R- product, which was the same as the SFC result, R-.

[0540] LCMS:[M+1] + =585, Purity: 98.5%

[0541] Preparation method of compound 2-P2

[0542]

[0543] At 25°C, lithium hydroxide (17 mg, 0.70 mmol) was added to a mixture of methyl 2-(((S)-2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azine-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate 2-6-P2 (80 mg, 0.13 mmol) in THF (5 mL) and H2O (1 mL). The mixture was stirred at 25°C for 3 hours. The reaction was detected by LCMS. The mixture was pre-purified by HPLC (0.05% NH3·H2O / ACN: 20%-35%) to give 2-(((S)-2-((4-cyano-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyrido[3,2-b][1,4]oxazin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid, compound 2-P2 (16.7 mg, 21%).

[0544] LCMS:[M+1] + =585, purity: 98.5%.

[0545] Example 3

[0546] 2-(((R)-2-((4-chloro-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid and 2-(((S)-2-((4-chloro-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (compounds 3-P1 and 3-P2)

[0547]

[0548] (1) Preparation of compound 3-2

[0549]

[0550] Sodium hydride (65 mg, 1.62 mmol) was added to a 10 mL solution of DMF (250 mg, 0.81 mmol) at 0 °C. Then 1-(bromomethyl)-4-chloro-2-fluorobenzene (172 mg, 0.81 mmol) was added. The mixture was stirred at room temperature for 3 hours under N2 atmosphere. The reaction was monitored by LCMS. The reaction mixture was quenched with ice water, extracted with EA, and the combined organic layers were washed with brine (30 mL), dried over Na2SO4, and concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography by elution with PE / EA (20:1) to give 2-((4-chloro-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azine-8(6H)-carboxylic acid tert-butyl ester (230 mg, 64%), as product 3-2.

[0551] LCMS: rt = 3.94 min, [M+1] + =450, purity: 93%.

[0552] (2) Preparation of compound 3-3

[0553]

[0554] HCl (3 mL) was added to a solution of 3-2 (230 mg, 0.53 mmol) in EA (5 mL). The mixture was stirred at 25 °C for 3 hours under N2 atmosphere. The reaction was monitored by LCMS. The reaction mixture was concentrated under reduced pressure to give 2-((4-chloro-2-fluorobenzyl)oxy)-6,6a,7,8,9,10-hexahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azine (180 mg, yield: 100%), product 3-3, as an HCl salt.

[0555] LCMS: rt = 2.4 min, [M+1] + =350, purity: 85%.

[0556] (3) Preparation of compounds 3-4

[0557]

[0558] At 25°C, 1-int-2 (155 mg, 0.52 mmol) was added to a mixture of 3-3 (180 mg, 0.52 mmol) and DIEA (340 mg, 2.60 mmol) in 10 mL of ACN. The mixture was stirred at 60°C for 7 hours under a nitrogen atmosphere. The reaction was detected by LCMS. The reaction mixture was quenched with water, extracted with EA, and the combined organic layers (30 mL) were washed with brine. The mixture was dried over Na2SO4 and concentrated under reduced pressure to obtain the residue, which was purified by silica gel column chromatography. The residue was eluted with PE / EA (1:5) to give 2-((2-((4-chloro-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]azine-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid ester (220 mg, 69%), as products 3-4. SFC showed a ratio of 1:1.

[0559] (4) Resolution of compounds 3-4

[0560]

[0561] The 3-4 sample (220 mg, 0.37 mmol) was further purified by SFC to obtain 3-4-P1 (90 mg, SFC rt = 2.47 min, yield: 41%) and 3-4-P2 (80 mg, SFC rt = 3.34 min, yield: 36%).

[0562] (5) Preparation of compounds 3-P1 and 3-P2

[0563]

[0564] At 25 °C, lithium hydroxide (18 mg, 0.75 mmol) was added to a mixture of methyl 3-4-P1 (90 mg, 0.15 mmol) in THF (5 mL) and H2O (1 mL). The mixture was stirred at 25 °C for 20 hours. The reaction was detected by LCMS, and the mixture was purified by pre-HPLC (0.05% NH3·H2O / ACN: 20%-35%) to give 2-(((R)-2-((4-chloro-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (18.5 mg, 50%).

[0565] LCMS: rt = 1.33 min, [M+1] + =594, Purity: 95%

[0566]

[0567] At 25 °C, lithium hydroxide (17 mg, 0.70 mmol) was added to a mixture of 3-4-P2 (80 mg, 0.13 mmol) in THF (5 mL) and H2O (1 mL). The mixture was stirred at 25 °C for 3 hours. The reaction was confirmed to be complete by LCMS. The mixture was purified by pre-HPLC (0.05% NH3·H2O / ACN: 20%-35%) to give 2-(((S)-2-((4-chloro-2-fluorobenzyl)oxy)-6a,7,9,10-tetrahydropyrazino[1,2-d]pyridino[3,2-b][1,4]oxazin-8(6H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (15.6 mg, 40%).

[0568] LCMS: rt = 1.34 min, [M+1] + =594, Purity: 96%

[0569] Example 4

[0570] 2-((2-((4-cyano-2-fluorobenzyl)oxy)-7a,8,10,11-tetrahydro-5H-pyrazino[2,1-c]pyridino[2,3-e][1,4]oxazapyro-9(7H)-yl)methyl)-1-(((S)-oxacyclobutane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (compound 4)

[0571]

[0572] (1) Preparation of compound 4-2

[0573]

[0574] DIPEA (2.24 g, 32.8 mmol, 2.0 eq) was added to a solution of 1-(tert-butyl)-3-methylpiperazine-1,3-dicarboxylic acid 4-1 (4.0 g, 16.4 mmol, 1 eq) and methyl 2,6-dichloronicotinate (3.38 g, 16.4 mmol, 1.0 eq) in NMP (50 mL). The mixture was stirred at 120 °C for 12 h under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was diluted with H2O (100 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were dried over Na2SO4, filtered and concentrated to obtain the residue, which was purified by silica gel column chromatography and eluted with (EA / PE = 0-20%) to give 4-(6-chloro-3-(methoxycarbonyl)pyridin-2-yl)-3-(2-methoxy-2-oxoethyl)piperazine-1-carboxylic acid tert-butyl ester 4-2 (1.6 g, 23.6% yield).

[0575] 1 H NMR (400MHz, CDCl3) δ7.96(t,J=7.1Hz,1H),6.75(dd,J=13.8,7.9Hz,1H),4.74(d,J=16.5Hz,1H),4.48(d,J=13.4Hz,1H),4.07–3.92(m ,1H),3.88(s,3H),3.75(s,3H),3.60(dd,J=19.9,9.7Hz,1H),3.48(d,J=13.0Hz,1H),3.37(d,J=11.4Hz,1H),3.15(s,1H),1.46(s,9H).

[0576] (2) Preparation of compound 4-3

[0577]

[0578] At 0 °C, LiAlH4 (441.6 mg, 11.616 mmol, 3.0 eq) was added to a THF (50 mL) solution of 4-2 (1.6 g, 3.9 mmol, 1.0 eq). The mixture was stirred at 0 °C for 10 min under N2. The reaction was monitored by LCMS. H2O (0.4 mL), 15% NaOH solution (0.4 mL), and EtOAc (50 mL) were slowly added to the mixture. The organic layer was washed with brine, dried over Na2SO4, and concentrated to give tert-butyl 4-(6-chloro-3-(hydroxymethyl)pyridin-2-yl)-3-(hydroxymethyl)piperazine-1-carboxylate 4-3 (640 mg, 46.3% yield).

[0579] LCMS: rt = 1.64 min, [M+1] + =358, Purity: 80%

[0580] (3) Preparation of compound 4-4

[0581]

[0582] Camphor sulfonic acid (3.26 g, 14 mmol, 5.0 eq) was added to a solution of 4-3 (1.0 g, 2.8 mmol, 1.0 eq) in toluene (150 mL). The mixture was stirred at 110 °C for 2 hours. The reaction was monitored by LCMS. The mixture was concentrated to obtain a residue, which was diluted with a saturated aqueous sodium bicarbonate solution. The residue was extracted with NaHCO3 (200 mL) and DCM (150 mL × 3). The organic layers were combined, dried over Na2SO4, filtered, concentrated, and the residue was purified by silica gel column chromatography, eluting with (MeOH / DCM = 0-10%) to give product 4-4 (366 mg, 54.7% yield).

[0583] LCMS: rt = 0.62 min, [M+1] + =240, Purity: 99.7%

[0584] (4) Preparation of compounds 4-5

[0585]

[0586] Add Boc₂O (390 mg, 1.78 mmol, 1.3 eq) and TEA (280 mg, 2.76 mmol, 2.0 eq) to a 15 mL solution of DCM containing 4-4 ​​(330 mg, 1.38 mmol, 1.0 eq). Stir the mixture at 10 °C for 2 hours. Monitor the reaction by LCMS. Concentrate the mixture to obtain a residue, which is purified by silica gel column chromatography, eluting with (EtOAc / PE = 0-10%) to give product 4-5 (360 mg, 76.9% yield).

[0587] LCMS: rt = 2.059 min, [M+1] + =340, Purity: 99.7%

[0588] (5) Preparation of compounds 4-6

[0589]

[0590] Under nitrogen atmosphere and stirring at 110 °C, 3-fluoro-4-(hydroxymethyl)benzylnitrile (259.03 mg, 1.711 mmol), Cs₂CO₃ (557.8 mg, 1.711 mmol), Xantphos (98.94 mg, 0.1711 mmol), and Pd₂(dba)₃ (78.3 mg, 0.0855 mmol) were added to 20 mL of dioxane solution 4-5 (290 mg, 0.855 mmol) for 60 hours. The reaction mixture was concentrated to a crude product and further purified by elution (EtOAC / PE = 0-20%) to give product 4-6 (347 mg, 89.4% yield).

[0591] LCMS: rt = 2.13 min, [M+1] + =455, Purity: 95%

[0592] (6) Preparation of compounds 4-7

[0593]

[0594] Add 4-6 (347 mg, 0.76 mmol) to 6 mL of HCl / EA solution and stir for 30 minutes at room temperature. Monitor the reaction by LCMS. Concentrate the reaction mixture to give crude product 4-7 (270 mg, 99% yield), which was used directly for the next step without purification.

[0595] LCMS: rt = 1.12 min, [M+1] + =335, Purity: 99%

[0596] (7) Preparation of compounds 4-8

[0597]

[0598] Under nitrogen atmosphere and stirring at room temperature, DIEA (920 mg, 0.71 mmol) was added to a solution of 4-7 (270 mg, 0.78 mmol) in 15 mL MeCN for 10 min. Then, 1-int-2 (209 mg, 0.71 mmol) was added to the reaction mixture at 60 °C for 16 h. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product and further purified by elution (EtOAc / PE = 0-5%) to give product 4-8 (210 mg, 60% yield).

[0599] LCMS: rt = 1.553 min, [M+1] + =613, Purity: 99%

[0600] (8) Preparation of Compound 4

[0601]

[0602] To a solution of 4-8 (200 mg, 0.32 mmol) in THF / H₂O (8 mL), the mixture was stirred at room temperature for 16 hours, and LiOH (40 mg, 1.63 mmol) was added. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by preparative HPLC to give compound 4 (70 mg, 75% yield).

[0603] LCMS: rt = 1.246 min, [M+1] + =599, Purity: 98%

[0604] (9) Preparation (resolution) of compounds 4-P1 and 4-P2

[0605]

[0606] The sample of compound 4 (70 mg, 0.117 mmol) was further purified by SFC to obtain 4-P1 (36.55 mg, SFC rt = 4.76 min, yield: 95%) and 4-P2 (9 mg, SFC rt = 5.9 min, yield: 26%), both of which were light white solids.

[0607] Compound 4-P1: LCMS: rt=1.246min, [M+1] + =599, Purity: 98%

[0608] 1¹H NMR (400 MHz, MeOD) δ 8.20 (s, 1H), 7.95 (dd, J = 8.4, 1.4 Hz, 1H), 7.65–7.52 (m, 4H), 7.38 (d, J = 8.0 Hz, 1H), 6.29 (d, J = 7.9 Hz, 1H), 5.45 (s, 2H), 5.28 (td, J = 6.9, 4.1 Hz, 1H), 4.83 (s, 1H), 4.71 (dd, J = 15.3, 2.9 Hz, 1H), 4.64 (dd, J = 13.1, 2.8 Hz, 2H), 4.56–4.41 (m, 2H), 3.93 (dt, J = 13.1, 12.0 Hz, 3H), 3.81 (dd, J = 12.9, 2.8 Hz, 1H), 3.71 (dd, J = 12.9, 6.3 Hz, 1H), 3.48–3.41 (m, 1H), 3.23 (t, J = 10.7 Hz, 1H), 2.81–2.73 (m, 3H), 2.56–2.46 (m, 1H), 2.36 (dd, J = 11.0, 8.4 Hz, 1H), 2.23 (t, J = 10.5 Hz, 1H).

[0609] Compound 4-P2: LCMS: r.t. = 1.274 min, [M+H] + = 599, purity: 98.7%.

[0610] 1 ¹H NMR (400 MHz, MeOD) δ 8.30 (s, 1H), 7.96 (d, J = 8.0 Hz, 1H), 7.67–7.52 (m, 4H), 7.38 (d, J = 8.0 Hz, 1H), 6.30 (d, J = 7.9 Hz, 1H), 5.46 (s, 2H), 5.25 (d, J = 4.8 Hz, 1H), 4.74–4.68 (m, 1H), 4.63 (t, J = 9.0 Hz, 2H), 4.56–4.43 (m, 2H), 4.01 (d, J = 13.7 Hz, 1H), 3.89 (t, J = 11.9 Hz, 2H), 3.79 (dd, J = 12.9, 2.8 Hz, 1H), 3.72–3.63 (m, 1H), 3.41 (s, 1H), 3.24 (s, 2H), 2.82 (t, J = 11.4 Hz, 2H), 2.69 (d, J = 10.6 Hz, 1H), 2.53 (s, 1H), 2.40 (s, 1H), 2.21 (t, J = 10.4 Hz, 1H).

[0611] Example 5

[0612] 2-(((R)-2-((4-cyano-2-fluorobenzyl)oxy)-6,7,7a,8,10,11-hexahydro-9H-pyrazino[1,2-d]pyrido[3,2-B][1,4]oxazono-9-yl)methyl)-1-(((S)-oxacyclobutane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid and 2-(((S)- 2-((4-cyano-2-fluorobenzyl)oxy)-6,7,7a,8,10,11-hexahydro-9H-pyrazino[1,2-d]pyrido[3,2-B][1,4]oxazono-9-yl)methyl)-1-(((S)-oxecyclobutane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (compounds 5-P1 and 5-P2)

[0613]

[0614] (1) Preparation of compound 5-2

[0615]

[0616] Under nitrogen atmosphere and stirring at 110 °C, 2,6-dichloro-3-fluoropyridine (3.04 g, 17.4 mmol), Cs₂CO₃ (12.6 g, 38.64 mmol), Xantphos (2.23 g, 3.864 mmol), and Pd₂(dba)₃ (1.77 g, 1.94 mmol) were added to 50 mL of a dioxane solution of 3-(2-hydroxyethyl)piperazine-1-carboxylic acid tert-butyl ester 5-1 (4000 mg, 17.4 mmol) for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by elution (MeOH / DCM = 0-5%) to give product 5-2 (3.6 g, 58% yield).

[0617] LCMS: rt = 1.17 min, [M+H] + =360, Purity: 90%

[0618] (2) Preparation of compound 5-3

[0619]

[0620] Under nitrogen atmosphere and at 1000 °C, NaH (711.4 mg, 17.78 mmol) was added to a 60 mL LDM solution of 5-2 (3.6 g, 8.89 mmol) for 4 hours, and the reaction was monitored by LCMS. The reaction mixture was poured into water (300 mL), extracted with EtOAc (100 mL × 3), washed with brine and dried, and concentrated to obtain the crude product. This crude product was further purified by elution (EtOAc / PE = 0-25%) to give product 5-3 (200 mg, yield 6.6%).

[0621] LCMS: rt = 2.148 min, [M+H] + =340, Purity: 95%

[0622] (3) Preparation of compound 5-4

[0623]

[0624] Under nitrogen atmosphere and at 110 °C, KOH (54.61 mg, 0.97 mmol), t-Buxphos (27.56 mg, 0.065 mmol), and Pd2(dba)3 (14.9 mg, 0.016 mmol) were added to 10 mL of dioxane / H2O solution of 5-3 (110 mg, 0.325 mmol), and the mixture was stirred for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to give a crude product, which was further purified by elution (EtOAc / PE = 0-70%) to give product 5-4 (80 mg, 65% yield).

[0625] LCMS: rt = 1.478 min, [M+H] + =322, Purity: 91%

[0626] (4) Preparation of compound 5-5

[0627]

[0628] Under nitrogen atmosphere and at room temperature, NaH (15 mg, 0.374 mmol) was added to 5 mL of DMF solution 5-4 (80 mg, 0.249 mmol) for 15 min with stirring. Then, under nitrogen atmosphere and at room temperature, 4-(bromomethyl)-3-fluorobenzyl nitrile (53.3 mg, 0.249 mmol) was added to the reaction mixture. The reaction mixture was monitored by LCMS. The reaction mixture was poured into water (50 mL), extracted with EtOAc (30 mL × 3), washed with brine and dried, and concentrated to give crude product. This crude product was further purified by elution (PE / EA = 0-13%) to give product 5-5 (90 mg, 76.9% yield).

[0629] LCMS: rt = 2.213 min, [M+H] + =455, Purity: 99%

[0630] (5) Preparation of compounds 5-6

[0631]

[0632] Add 5 mL of HCl / EA solution to 5-5 (90 mg, 0.198 mmol) and stir for 30 minutes at room temperature. Monitor the reaction by LCMS. Concentrate the reaction mixture to give crude product 5-6 (70 mg, 98%), which was used directly in the next step without purification.

[0633] LCMS: rt = 1.174 min, [M+H] + =355, Purity: 87%

[0634] (6) Preparation of compounds 5-7

[0635]

[0636] Under nitrogen atmosphere and with stirring at room temperature for 10 minutes, DIEA (255.9 mg, 1.98 mmol) was added to a solution of 5-6 (70 mg, 0.198 mmol) in 10 mL of MeCN. Then, 1-int-2 (58.14 mg, 0.198 mmol) was added to the reaction mixture at 60 °C for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product and further purified by elution (EtOAc / PE = 0-5%) to give 5-7 (80 mg, 66% yield) as a white solid.

[0637] LCMS: rt = 1.513 min, [M+H] + =613, Purity: 88%

[0638] (7) Resolution and preparation of compounds 5-P1 and 5-P2

[0639]

[0640] LiOH (5.5 mg, 0.23 mmol) was added to a solution of 5-7-P2 (28 mg, 0.045 mmol) in THF / H2O (4 mL), and the mixture was stirred at room temperature for 16 hours, with the reaction monitored by LCMS. The reaction mixture was concentrated to a crude product and further purified by preparative HPLC to give compound 5-P2 (13.65 mg, 69.6%).

[0641] LCMS: rt = 1.246 min, [M+H]+ =599, Purity: 98%

[0642]

[0643] LiOH (5.5 mg, 0.23 mmol) was added to a solution of 5-7-P1 (28 mg, 0.045 mmol) in THF / H2O (4 mL), and the mixture was stirred at room temperature for 16 hours, with the reaction monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by preparative HPLC to give 5-P1 (7.25 mg, 26.6%) as a white solid.

[0644] LCMS: rt = 1.246 min, [M+H] + =599, Purity: 98%

[0645] Example 6

[0646] (S)-2-((2-((4-cyano-2-fluorobenzyl)oxy)-5,8,10,11-tetrahydrooxohexazo[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazol-6-carboxylic acid (compound 6)

[0647]

[0648] (1) Preparation of compound 6-2

[0649]

[0650] Sulfuric acid (10 mL) was added to a mixture of 2-chloro-6-hydroxynicotinic acid 6-1 (5.0 g, 29 mmol) in MeOH (40 mL). The mixture was stirred at 80 °C for 16 hours. The reaction was detected by LCMS. The reaction mixture was quenched with ice water, extracted with EA, and the combined organic layers (50 mL) were washed with brine, dried over Na2SO4, and concentrated under reduced pressure to give methyl 2-chloro-6-hydroxynicotinic acid 6-2 (4.5 g, 83%).

[0651] LCMS: rt = 2.1 min, [M+H] + =188, Purity: 92%

[0652] (2) Preparation of compound 6-3

[0653]

[0654] The reaction mixture of 6-2 (3.4 g, 18.2 mmol), PMB-Cl (3.4 g, 21.2 mmol), and K2CO3 (3.76 g, 27.3 mmol) in DMF (50 mL) was added. The mixture was stirred with Ar2 at 80 °C for 2 hours. The reaction was detected by LCMS. The reaction mixture was quenched by adding water. The aqueous phase was extracted with EtOAc (100 mL × 3) and washed with brine (50 mL × 2). The combined organic layers were dried over Na2SO4. The mixture was concentrated and purified by elution (PE / EA = 0-20%) to give methyl 2-chloro-6-((4-methoxybenzyl)oxy)nicotinic acid 6-3 (2.9 g, 52%).

[0655] LCMS: rt = 3.5 min, [M+H] + =308, purity: 95%

[0656] (3) Preparation of compound 6-4

[0657]

[0658] Add 6-3 (2.9 g, 9.4 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxabortazacyclopentan-2-yl)-5,6-dihydropyridine-1,3(2H)-dicarboxylic acid 1-4 methyl ester (4.16 g, 11.3 mmol), C s2 CO3 (6.16 g, 18.8 mmol) and Pd(dppf)Cl2 (0.69 g, 0.94 mmol) were reacted in 1,4-dioxane (80 mL). The mixture was stirred with Ar2 at 110 °C for 16 h. The reaction was monitored by LCMS. The mixture was concentrated and purified by elution (PE / EA = 0–20%) to give 1'-(tert-butyl)3,3'-dimethyl-6-((4-methoxybenzyl)oxy)-5',6'-dihydro-[2,4'-bipyridine]-1',3,3'(2'H)-tricarboxylic acid ester 6-4 (2.5 g, 53.2%).

[0659] LCMS: rt = 1.37 min, [M+H] + =513, Purity: 95%

[0660] (4) Preparation of compound 6-5

[0661]

[0662] 6-4 (1.5 g, 2.9 mmol) was dissolved in anhydrous THF (15 mL), and then LiAlH4 (0.222 g, 5.8 mmol) was added in portions at 0 °C. The reaction was detected by LCMS after 10 minutes. The reaction mixture was quenched with ice water (0.5 mL). The mixture was filtered and concentrated to give tert-butyl 3,5'-di(hydroxymethyl)-6-((4-methoxybenzyl)oxy)-3',6'-dihydro-[2,4'-bipyridine]-1'(2'H)-carboxylic acid 6-5 (1.2 g, 92%).

[0663] LCMS: rt = 1.95 min, [M+H] + =457, Purity: 88%

[0664] (5) Preparation of compound 6-6

[0665]

[0666] The reaction mixture of 6-5a (1.1 g, 2.4 mmol) and camphor sulfonic acid (3.24 g, 9.6 mmol) in TOL (20 mL) was added. The mixture was heated at 110 °C for 1 hour. The reaction was detected by LCMS. The mixture was concentrated and purified by elution (MeOH / DCM = 0-20%) to give 5,7,8,9,10,11-hexahydroxyoxetane-heptatrien-[4,3-b:6,5-c']bipyridin-2-ol 6-6 (0.4 g, 76%).

[0667] LCMS: rt = 1.25 min, [M+H] + =219, Purity: 96%

[0668] (6) Preparation of compound Int-5

[0669]

[0670] A solution of 6-6 (0.35 g, 1.6 mmol), (BOC)₂O (0.42 g, 1.9 mmol), and TEA in DCM (15 mL) was prepared. The mixture was stirred at room temperature for 1 hour. The reaction was detected by LCMS. The solution was concentrated and purified by elution (MeOH / DCM = 0–10%) to give 2-hydroxy-5,8,10,11-tetrahydrooxetane-heptatrienone [4,3-b:6,5-c']bipyridine-9(7H)-carboxylate Int-5 (350 mg, 70%).

[0671] LCMS: rt = 2.32 min, [M+H] + =319, Purity: 95%

[0672] 1H NMR (400MHz, CDCl3) δ12.63(s,1H),7.43(d,J=9.2Hz,1H),6.49(d,J=9.1Hz,1 H),4.17(s,4H),3.80(s,2H),3.67(t,J=5.5Hz,2H),2.72(s,2H),1.51(s,9H).

[0673] (7) Preparation of compounds 6-7

[0674]

[0675] Int-5 (0.3 g, 1.2 mmol) was dissolved in anhydrous DMF (5 mL), and then NaH (45 mg, 1.44 mmol) was added in portions at 0 °C. After 5 minutes, a solution of 4-(bromomethyl)-3-fluorobenzyl nitrile (0.202 g, 1.2 mmol) (5 mL DMF) was added to the reaction mixture through a sleeve. The reaction was monitored by LCMS after 20 minutes. The reaction mixture was quenched by adding water. The aqueous phase was extracted with EtOAc (20 mL × 3) and washed with brine (20 mL × 2). The combined organic layers were dried over Na₂SO₄. Concentrated and purified by elution (PE / EA = 0-20%), yielding 2-((4-cyano-2-fluorobenzyl)oxy)-5,8,10,11-tetrahydrooxetrazine-heptanetrien[4,3-b:6,5-c']bipyridine-9(7H)-carbamate 6-7 (0.35 g, 83.3%).

[0676] (8) Preparation of compounds 6-8

[0677]

[0678] Add 6-7 (0.35 g, 0.78 mmol) to HCl / EA (20 mL, 3 M). Stir the mixture at room temperature for 0.5 h. Analyze the reaction by LCMS. Concentrate the mixture to give 3-fluoro-4-(((5,7,8,9,10,11-hexahydroxyoxetanetrien[4,3-b:6,5-c']dipyridin-2-yl)oxy)methyl)benzyl nitrile 6-8 (0.27 g, 98%).

[0679] LCMS: rt = 2.21 min, [M+H] + =351, Purity: 95%

[0680] (9) Preparation of compound 6-A

[0681]

[0682] The reaction mixture of 6-8 (0.27 g, 1.1 mmol) and DIEA (0.451 g, 3.5 mmol) in 20 mL of CH3CN was stirred at room temperature for 10 min. Then, 1-int-2 (206.5 g, 1.0 mmol) was added, and the mixture was heated at 65 °C for 15 h. The reaction was monitored by LCMS. The product was concentrated and purified by elution (MeOH / DCM = 0-8%) to give compound 6-A (250 mg, 59%).

[0683] LCMS: rt = 2.51 min, [M+H] + =610, Purity: 95%

[0684] 1 H NMR (400MHz, CDCl3) δ8.16(s,1H),7.99(dd,J=8.5,1.3Hz,1H),7.77(d,J=8.5Hz,1H),7.60(t,J= 7.5Hz,1H),7.51(d,J=8.2Hz,1H),7.40(dd,J=28.4,9.0Hz,2H),6.70(d,J=8.2Hz,1H),5.51(s,2 H),5.26–5.17(m,1H),4.76–4.58(m,3H),4.42–4.33(m,3H),4.18–4.09(m,2H),3.95(s,3H),3.8 8(s,2H),3.25(s,2H),2.81(t,J=5.5Hz,2H),2.72(p,J=8.2Hz,3H),2.45(dq,J=11.2,7.3Hz,1H).

[0685] (10) Preparation of compound 6

[0686]

[0687] 6-A (0.25 g, 0.41 mmol) was dissolved in THF (4 mL), and then an aqueous lithium hydroxide solution (4 mL) was added. The mixture was stirred at room temperature for 8 hours. The reaction was detected by LCMS. The solution was concentrated and purified by preparative HPLC (NH3·H2O) to give compound 6 (0.13 g, 53%). LCMS: rt = 1.225 min, [M+H] + =596, Purity: 99%

[0688] 1H NMR(400MHz,MeOD)δ8.17(s,1H),7.95(dd,J=8.4,1.5Hz,1H),7.69–7.52(m,5H),6.75(d,J= 8.2Hz,1H),5.54(s,2H),5.26(d,J=4.2Hz,2H),4.70(dd,J=15.4,3.1Hz,1H),4.61(dd,J=13 .7,7.8Hz,1H),4.45(dt,J=9.1,5.9Hz,1H),4.35(s,2H),4.10(dd,J=34.7,13.6Hz,2H),3.8 7(s,2H),3.48(d,J=1.7Hz,1H),3.24–3.23(m,1H),2.85–2.71(m,5H),2.53(d,J=8.8Hz,1H).

[0689] (11) Preparation (resolution) of compound 6-P1

[0690]

[0691] Compound 6 (22 mg, 0.037 mmol) was dissolved in H₂O (2 mL), and then NaOH (1.48 mg, 0.037 mmol) was added. The mixture was stirred at room temperature for 2 minutes. The solution was concentrated to give compound 6-P1 (22.8 mg, 100%).

[0692] LCMS: rt = 1.225 min, [M+H] + =596, Purity: 96%

[0693] Example 7

[0694] (S)-2-((2-((4-cyano-2-fluorobenzyl)oxy)-5,8,10,11-tetrahydrooxoazo[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-3-(oxetane-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylic acid (compound 7)

[0695]

[0696] (1) Preparation of compound 7-A

[0697]

[0698] The reaction mixture of 6-8 (96 mg, 0.27 mmol), 1-int-2 (80 mg, 0.27 mmol), and N,N-diisopropylethylamine (175 mg, 1.35 mmol) in 5 mL of acetonitrile was stirred at 60 °C for 16 h. The reaction was monitored by TLC. The reaction mixture was evaporated to dryness, and the residue was purified by SGC (EA / MeOH = 10:1) to give compound 7-A (100 mg, 60%).

[0699] LCMS: rt = 0.773 min, [M+H]+ = 611, purity: 97%

[0700] (2) Preparation of compound 7

[0701]

[0702] At 25 °C, 1 mL of a lithium hydroxide (20 mg, 0.81 mmol) H₂O solution was added to a 5 mL reaction mixture of compound 7-A (100 mg, 0.16 mmol) and tetrahydrofuran. The mixture was stirred at 25 °C for 3 hours. The reaction mixture was monitored by LCMS until completion. The reaction mixture was purified by pre-HPLC to give compound 7 (22.45 mg, 22%).

[0703] LCMS: rt = 1.211 min, [M+H] + =597, Purity: 99%

[0704] 1 H NMR (400MHz, CDCl3) δ8.19(q,J=8.2Hz,2H),7.61(t,J=7.5Hz,1H),7.52(d,J=8.3Hz,1H ),7.44(d,J=7.8Hz,1H),7.37(d,J=9.1Hz,1H),6.71(d,J=8.2Hz,1H),5.51(s,2H),5.25 (s,1H),4.93–4.85(m,1H),4.79(d,J=12.9Hz,1H),4.61(d,J=7.1Hz,1H),4.35(s,3H), 4.20(s,2H),3.87(s,2H),3.28(s,2H),2.84(d,J=5.4Hz,2H),2.74(s,3H),2.46(s,1H).

[0705] Example 8

[0706] (S)-2-((2-((2-fluoro-4-(trifluoromethyl)benzyl)oxy)-5,8,10,11-tetrahydrooxohexazo[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazol-6-carboxylic acid (compound 8)

[0707]

[0708] (1) Preparation of compound 8-7

[0709]

[0710] 60% sodium hydride (19 mg, 0.75 mmol) was slowly added to Int-5 (200 mg, 0.62 mmol) in 4 mL of N,N-dimethylformamide at 0 °C. The mixture was stirred at 0 °C for 15 min. Then, 1-(bromomethyl)-2-fluoro-4-(trifluoromethyl)benzene (162 mg, 0.62 mmol) was added to the mixture. The reaction was monitored by TLC. Then, an ammonium chloride solution was added to the mixture, and the mixture was extracted with ethyl acetate. The organic phase was evaporated to dryness, and the residue was purified by SGC (PE:EA = 5:1) to give compound 8-7 (150 mg, 48%).

[0711] LCMS: rt = 1.12 min, [M+H] + =495, Purity: 95%

[0712] (2) Preparation of compound 8-8

[0713]

[0714] 8-7 (150 mg, 0.30 mmol) was added to 6 mL of (3 M) ethyl acetate solution of hydrogen chloride and stirred at 25 °C for 2 hours. The reaction was monitored by TLC. The reaction mixture was evaporated to dryness to give crude product 8-8 (120 mg, 99%).

[0715] LCMS: rt = 0.764 min, [M+H] + =395, Purity: 51%

[0716] (3) Preparation of compounds 8-9

[0717]

[0718] A solution of 8-8 (90 mg, 0.30 mmol), 1-int-2 (120 mg, 0.30 mmol), and N,N-diisopropylethylamine (197 mg, 1.52 mmol) in 5 mL of acetonitrile was stirred at 60 °C for 16 hours. The reaction was detected by TLC. The reaction mixture was evaporated to dryness, and the residue was purified by SGC (PE:EA = 1:1) to give compound 8-9 (60 mg, 30%).

[0719] LCMS: rt = 1.73 min, [M+H] + =653, Purity: 91%

[0720] (4) Preparation of compound 8

[0721]

[0722] Compound 8-9 (60 mg, 0.09 mmol) was added to a reaction mixture in 5 mL of tetrahydrofuran at 25 °C. The reaction mixture was stirred at 25 °C for 16 h. The reaction was detected by LCMS. The reaction mixture was purified by pre-HPLC to give compound 8 (8.3 mg, 14%) as (S)-2-((2-((2-fluoro-4-(trifluoromethyl)benzyl)oxy)-5,8,10,11-tetrahydrooxo-hexazo[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid.

[0723] LCMS: rt = 0.887 min, [M+H] + =639, Purity: 99%

[0724] 1 H NMR (400MHz, CDCl3) δ8.19(s,1H),8.04(d,J=8.3Hz,1H),7.81(d,J=8.3Hz,1H),7.61(t,J =7.0Hz,1H),7.51(d,J=7.8Hz,1H),7.40(d,J=7.7Hz,1H),7.33(d,J=9.5Hz,1H),6.70(d,J =8.1Hz,1H),5.51(s,2H),5.21(s,1H),4.78–4.59(m,3H),4.36(d,J=12.7Hz,3H),4.15(s ,2H),3.88(s,2H),3.28(s,2H),2.85(s,2H),2.77(s,2H),2.73–2.67(m,1H),2.45(s,1H).

[0725] Example 9

[0726] 2-(((6a,10a)-2-((4-cyano-2-fluorobenzyl)oxy)-5,6a,7,9,10,10a-hexahydro-8H-pyrano[4,5-b:2,3-c']bipyridin-8-yl)methyl)-1-(((S)-oxecyclobutane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (compound 9)

[0727]

[0728] (1) Preparation of compounds 9-A and 9-B

[0729]

[0730] At 25 °C, LiOH (26 mg, 1.08 mmol, 5 eq) in 1 mL of water was added to a THF solution of 2-(((6aR,10aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,6a,7,9,10,10a-hexahydro-8H-pyrano[4,5-b:2,3-c']bipyridin-8-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (130 mg, 0.22 mmol, 1 eq) in 4 mL of water. The mixture was stirred for 16 hours, and the reaction was monitored by LCMS. The reactants were concentrated, pre-HPLC purified, and lyophilized to give 2-(((6aR,10aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,6a,7,9,10,10a-hexahydro-8H-pyrano[4,5-b:2,3-c']bipyridin-8-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (33.39 mg, 26.3%). LCMS: rt = 1.232 min, [M+H] + =584.2, Purity: 100%

[0731] At 25 °C, LiOH (26 mg, 1.08 mmol, 5 eq) in 1 mL of water was added to a THF solution of 2-(((6aS,10aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,6a,7,9,10,10a-hexahydro-8H-pyrano[4,5-b:2,3-c']bipyridin-8-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (130 mg, 0.22 mmol, 1 eq) in 4 mL of water. The mixture was stirred for 16 hours, and the reaction was monitored by LCMS. The reactants were concentrated, pre-HPLC purified and lyophilized to give 2-(((6aS,10aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,6a,7,9,10,10a-hexahydro-8H-pyrano[4,5-b:2,3-c']bipyridin-8-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (49.99 mg, 39.4%).

[0732] LCMS: rt = 1.240 min, [M+H] + =584.2, Purity: 100%

[0733] Example 10

[0734] (S)-2-((2-((4-chloro-2-fluorobenzyl)oxy)-5,8,10,11-tetrahydrooxohepteno[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazol-6-carboxylic acid (compound 10)

[0735]

[0736] (1) Preparation of compound 10-7

[0737]

[0738] At 0°C, NaH (38 mg, 1.57 mmol, 1.5 eq) was added to a DMF (8 mL) solution of Int-5 (200 mg, 0.63 mmol, 1 eq) for 30 minutes. 1-(bromomethyl)-4-chloro-2-fluorobenzene (140 mg, 0.63 mmol, 1.0 eq) was added, and the mixture was stirred at 25°C for 30 minutes. The reaction was monitored by TLC. The mixture was diluted with EtOAc (50 mL), washed with H2O (70 mL), and extracted with EtOAc (50 mL × 2). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the residue, which was purified by silica gel column chromatography, eluting with (PE / EA = 0-20%) to give product 10⁻⁷ (230 mg, 79.6% yield).

[0739] 1 H NMR (400MHz, CDCl3) δ7.53(d,J=8.3Hz,1H),7.43(t,J=8.1Hz,1H),7.16–7.04(m,2H),6.69(d,J=8.2Hz,1H),5 .43(s,2H),4.34(s,2H),4.12(d,J=2.9Hz,2H),3.88(s,2H),3.64(t,J=5.4Hz,2H),2.76(s,2H),1.51(s,9H).

[0740] (2) Preparation of compound 10-8

[0741]

[0742] Add 10⁻⁷ (200 mg, 0.43 mmol, 1 eq) to a HCl / EA (6 mL) solution. Stir the reaction mixture at 25 °C for 1 hour. Monitor the reaction mixture by LC-MS. Concentrate the reaction mixture under reduced pressure to give 10⁻⁸ (160 mg, crude product).

[0743] LCMS: rt = 1.921 min, [M+H] + =361.1, Purity: 95.2%

[0744] (3) Preparation of compound 10-9

[0745]

[0746] DIEA (286 mg, 2.2 mmol, 5 eq) was added to a solution of 10⁻⁸ (160 mg, 0.44 mmol, 1 eq) and 1-int⁻² (130 mg, 0.44 mmol, 1 eq) in MeCN (6 mL). The mixture was stirred at 60 °C for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue, which was purified by silica gel column chromatography, eluting with (MeOH / DCM = 0-10%), to give product 10⁻⁹ (150 mg, 55% yield).

[0747] LCMS: rt = 1.553 min, [M+H] + =619.4, Purity: 94.7%

[0748] (4) Preparation of compound 10

[0749]

[0750] At 25 °C, a solution of 10⁻⁹ (150 mg, 0.2 mmol, 1 eq) of THF (5 mL) was added to a solution of LiOH (29 mg, 1.0 mmol, 5 eq) in H₂O (1 mL). The mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated and purified by HPLC, and lyophilized to give (S)-2-((2-((4-chloro-2-fluorobenzyl)oxy)-5,8,10,11-tetrahydrooxetrazine-hepteno[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid, compound 10 (57.53 mg, 39.2% yield).

[0751] LCMS: rt = 1.316 min, [M+H] + =605.2, Purity: 100%

[0752] 1H NMR (400MHz, CD3OD_SPE) δ8.17(d,J=0.8Hz,1H),7.95(dd,J=8.4,1.5Hz,1H),7.59(d,J=8.5Hz,2H),7.47( t,J=8.1Hz,1H),7.26–7.10(m,2H),6.69(d,J=8.2Hz,1H),5.42(s,2H),5.26(qd,J=7.1,2.9Hz,1H),4.88( d,J=7.1Hz,1H),4.70(dd,J=15.3,3.0Hz,1H),4.63–4.56(m,1H),4.45(dt,J=9.1,6.0Hz,1H),4.34(s,2H) ,4.10(dd,J=37.2,13.6Hz,2H),3.86(s,2H),3.23(t,J=10.8Hz,2H),2.85–2.70(m,5H),2.57–2.47(m,1H).

[0753] Example 11

[0754] (S)-2-((2-((4-cyano-2-fluorobenzyl)oxy)-5,8,10,11-tetrahydrooxetrazine-hepteno[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (compound 11)

[0755]

[0756] (1) Preparation of compound 11-7

[0757]

[0758] Int-5 (0.3 g, 1.2 mmol) was dissolved in anhydrous DMF (5 mL), and then NaH (45 mg, 1.44 mmol) was added in portions at 0 °C. After 5 minutes, a solution of 4-(bromomethyl)-3-fluorobenzyl nitrile (0.202 g, 1.2 mmol) (5 mL DMF) was added to the reaction mixture through a sleeve. The reaction was detected by LC-MS after 20 minutes. The reaction mixture was quenched by adding water. The aqueous phase was extracted with EtOAc (20 mL × 3) and washed with brine (20 mL × 2). The combined organic layers were dried over Na₂SO₄. The product was concentrated and purified by elution (PE / EA = 0-20%) to give product 11-7 (0.35 g, 83.3%).

[0759] LCMS: rt = 2.21 min, [M+H] + =477.5, Purity: 95%

[0760] (2) Preparation of compound 11-8

[0761]

[0762] Add 11-7 (0.35 g, 0.78 mmol) to HCl / EA (20 mL, 3 M). Stir the mixture at room temperature for 0.5 hours. Analyze the reaction by LC-MS. Concentrate the mixture to give product 11-8 (0.27 g, 98%).

[0763] LCMS: rt = 1.21 min, [M+H] + =376.2, Purity: 92%

[0764] (3) Preparation of compound 11-9

[0765]

[0766] The reaction mixture of 11-8 (0.27 g, 1.1 mmol) and DIEA (0.451 g, 3.5 mmol) in 20 mL of CH3CN was stirred at room temperature for 10 min. Then, 1-int-2 (206.5 g, 1.0 mmol) was added, and the mixture was heated at 65 °C for 15 h. The reaction was detected by LC-MS. The product was concentrated and purified by elution (MeOH / DCM = 0-8%) to give product 11-9 (250 mg, 59%).

[0767] LCMS: rt = 2.51 min, [M+H] + =635, Purity: 95%

[0768] 1 H NMR (400MHz, CDCl3) δ8.16(s,1H),7.99(dd,J=8.5,1.3Hz,1H),7.77(d,J=8.5Hz,1H),7.60(t,J= 7.5Hz,1H),7.51(d,J=8.2Hz,1H),7.40(dd,J=28.4,9.0Hz,2H),6.70(d,J=8.2Hz,1H),5.51(s,2 H),5.26–5.17(m,1H),4.76–4.58(m,3H),4.42–4.33(m,3H),4.18–4.09(m,2H),3.95(s,3H),3.8 8(s,2H),3.25(s,2H),2.81(t,J=5.5Hz,2H),2.72(p,J=8.2Hz,3H),2.45(dq,J=11.2,7.3Hz,1H).

[0769] (4) Preparation of compound 11

[0770]

[0771] 11-9 (0.25 g, 0.41 mmol) was dissolved in THF (4 mL), and then an aqueous lithium hydroxide solution (4 mL) was added. The mixture was stirred at room temperature for 8 hours. The reaction was detected by LC-MS. The solution was concentrated and purified by preparative HPLC (NH3·H2O) to give (S)-2-((2-(((4-cyano-2-fluorobenzyl)oxy)-5,8,10,11-tetrahydrooxetane-hepteno[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(oxetane-2-ylmethyl)-1H-benzo[d]imidazolium-6-carboxylic acid (0.13 g, 53%).

[0772] LCMS: rt = 1.225 min, [M+H] + =621, Purity: 96%

[0773] Example 12

[0774] 2-(((7a,11a)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetaneheptan[4,3-B:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (compound 12)

[0775]

[0776] (1) Preparation of compound 12-7

[0777]

[0778] 13-6 (190 mg, 594 mmol) was added to 10 mL of DMF under nitrogen and stirring at 25 °C, along with 4-(bromomethyl)-3-fluorobenzyl nitrile (127.1 mg, 0.594 mmol) and NaH (35.60 mg, 0.89 mmol) for 0.5 h. The reaction was monitored by LCMS. The reaction mixture was poured into water (30 mL), extracted with DCM (20 mL × 3), washed with brine and dried, and concentrated to give the crude product. This crude product was further purified by elution with EtOAc / (PE = 0-47%) to give product 12-7 (260 mg, yield: 70%).

[0779] LCMS: rt = 2.109 min, [M+H] + =454, Purity: 98%

[0780] (2) Preparation of compound 12-8

[0781]

[0782] A solution of 12-7 (260 mg, 0.57 mmol) in 6 mL of HCl / EA was stirred at room temperature for 30 minutes. The reaction was monitored by LCMS. The reaction mixture was concentrated to give crude product 12-8 (200 mg), which was used directly in the next step without purification.

[0783] LCMS: rt = 0.88 min, [M+H] + =355, Purity: 88%

[0784] (3) Preparation of compound 12-9

[0785]

[0786] Under nitrogen atmosphere, at room temperature and with stirring, DIEA (736.7 mg, 0.57 mmol) was added to a solution of 12-8 (200 mg, 0.57 mmol) in 15 mL MeCN for 10 min. Then, 1-int-2 (166.6 mg, 5.7 mmol) was added to the reaction mixture at 60 °C for 16 h. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by elution (PE / EtOAc = 0-5%) to give product 12-9 (200 mg, yield: 57.8%).

[0787] LCMS: rt = 1.553 min, [M+H] + =613, Purity: 98%

[0788] (4) Resolution of compound 12-9

[0789]

[0790] 12-9 (180 mg, 0.327 mmol) was further purified by SFC to obtain 2-(((7aS,11aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(((S)-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (80 mg, SFC) rt=3.376mins, yield: 88%) and 2-(((7aR,11aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(((S)-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (80 mg, SFC rt=3.926mins, yield: 88%).

[0791] (5) Preparation of compounds 12-A and 12-B

[0792]

[0793] LiOH (15.7 mg, 0.655 mmol) was added to a solution of 2-(((7aS,11aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(((S)-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (80 mg, 0.131 mmol) in THF / H2O (5 mL), and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by preparative HPLC to give 2-(((7aS,11aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(((S)-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (50.6 mg, yield: 65%).

[0794] LCMS: rt = 1.242 min, [M+H] + =599, purity 100%

[0795] LiOH (15.7 mg, 0.655 mmol) was added to a solution of 2-(((7aR,11aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(((S)-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (80 mg, 0.131 mmol) in THF / H2O (5 mL), and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by preparative HPLC to give 2-(((7aR,11aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-(((S)-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (45.6 mg, yield: 60%).

[0796] LCMS: rt = 1.226 min, [M+H] + =599, purity 100%

[0797] Example 13

[0798] 2-(((7aS,11aS)-2-((4-(difluoromethyl)-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid and

[0799] 2-(((7aR,11aR)-2-((4-(difluoromethyl)-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (compound 13A / B)

[0800]

[0801] (1) Preparation of compound 13-1

[0802]

[0803] To a solution of methyl 2-chloro-6-methoxynicotinate (2 g, 10 mmol, 1.0 eq) in dioxane (50 mL), 1-(tert-butyl)-3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-5,6-dihydropyridine-1,3(2H)-dicarboxylic acid ester (3.67 g, 10 mmol, 1.0 eq), K₂CO₃ (2.76 g, 20 mmol, 2.0 eq), and Pd(dppf)Cl₂ (731 mg, 1 mmol, 0.1 eq) were added. The mixture was stirred at 90 °C for 16 hours under a nitrogen atmosphere. The reaction was monitored by LCMS. The mixture was directly concentrated to give a residue, which was purified by silica gel column chromatography by elution with (EA / P = 0-30%) to give product 13-1 (1.1 g, 27.1% yield).

[0804] LCMS: rt = 2.025 min, [M-55] + =407, Purity: 92%

[0805] (2) Preparation of compound 13-2

[0806]

[0807] At 0 °C, LiAlH4 (152 mg, 4 mmol, 2.0 eq) was added to a THF (50 mL) solution of 13-1 (0.83 g, 2 mmol, 1.0 eq), and the mixture was stirred at 20 °C for 10 min. The reaction was monitored by LCMS. H2O (0.2 mL), 15% NaOH solution (0.2 mL), and EtOAc (50 mL) were slowly added to the mixture. The organic layer was washed with brine, dried over Na2SO4, and concentrated to give product 13-2 (630 mg, 90% yield).

[0808] LCMS: rt = 1.57 min, [M+H] + =351.0, Purity: 90%

[0809] (3) Preparation of compound 13-3

[0810]

[0811] A solution of 13-2 (300 mg, 0.286 mmol, 1.0 eq) and Pd / C (100 mg) in THF (15 mL) was stirred at 20 °C for 14 hours under H2 (15 psi). The reaction was monitored by LCMS. The resulting mixture was filtered, and the filter cake was washed with THF (3 × 20 mL). The filtrate was directly concentrated to obtain the residue, which was purified by column chromatography on silica gel, eluting with (PE / EA = 0-50%) to give product 13-3 (118 mg, 40% yield).

[0812] LCMS: rt = 1.60 min, [M+H] + =353.2, Purity: 95.9%

[0813] (4) Preparation of compound 13-4

[0814]

[0815] Camphor sulfonic acid (3.95 g, 17 mmol, 5.0 eq) was added to a solution of 13-3 (1.2 g, 3.4 mmol, 1.0 eq) in toluene (80 mL). The mixture was stirred at 110 °C for 2 hours. The reaction was monitored by LCMS. The reaction mixture was directly concentrated to obtain a residue, which was purified by silica gel column chromatography by elution with (MeOH / DCM = 0-10%) to give product 13-4 (700 mg, 87.9% yield).

[0816] LCMS: rt = 1.528 min, [M+H] + =235.2, Purity: 86.9%

[0817] (5) Preparation of compound 13-5

[0818]

[0819] A solution of 3-4 (0.35 g, 1.5 mmol, 1.0 eq) in 10 mL of HBr was stirred at 120 °C for 6 hours. The reaction was monitored by LC-MS. The reaction mixture was adjusted to pH 7 with NaOH (1N) and extracted with DCM (20 mL × 3). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to give product 13-5 (200 mg, 60.6% yield).

[0820] LCMS: rt = 0.385 min, [M+H] + =221.1, Purity: 87.18%

[0821] (6) Preparation of compound 13-6

[0822]

[0823] TEA (184.2 mg, 1.82 mmol, 2.0 eq) and Boc₂O (238 mg, 1.1 mmol, 1.2 eq) were added to a 10 mL solution of DCM containing 13-5 (200 mg, 0.91 mmol, 1.0 eq). The mixture was stirred at 25 °C for 2 hours. The reaction was monitored by LCMS. The mixture was directly concentrated to obtain a residue, which was purified by silica gel column chromatography by elution with (MeOH / DCM = 0-10%) to give product 13-6 (190 mg, 65.5% yield). LCMS: rt = 1.35 min, [M+H] + =343, Purity: 93.8%

[0824] (7) Preparation of compound 13-7

[0825]

[0826] To 13-6 (190 mg, 594 mmol), 1-(bromomethyl)-4-(difluoromethyl)-2-fluorobenzene (127.1 mg, 0.594 mmol) and NaH (35.60 mg, 0.89 mmol) were added to 10 mL of LDM, and the mixture was stirred at 25 °C for 0.5 h under nitrogen atmosphere. The reaction was monitored by LCMS. The reaction mixture was poured into water (30 mL), extracted with DCM (20 mL × 3), washed with brine and dried, and concentrated to give a crude product. This crude product was further purified by elution (PE / EtOAc = 0-47%) to give product 13-7 (260 mg, yield: 70%).

[0827] LCMS: rt = 2.323 min, [M-55] + =423, Purity: 99.7%

[0828] (8) Preparation of compound 13-8

[0829]

[0830] 13-7 (260 mg, 0.57 mmol) was stirred in 6 mL HCl / EA at room temperature for 30 minutes. The reaction was monitored by LCMS. The reaction mixture was concentrated to give crude product 13-8 (200 mg), which was used directly in the next step without purification.

[0831] LCMS: rt = 0.846 min, [M+H] + =376, Purity: 95%

[0832] (9) Preparation of compounds 13-9

[0833]

[0834] Under nitrogen atmosphere, DIEA (736.7 mg, 0.57 mmol) was added to 15 mL of MeCN containing 13-8 (200 mg, 0.57 mmol), and the mixture was stirred at room temperature for 10 minutes. Then, 1-int-2 (166.6 mg, 5.7 mmol) was added to the reaction mixture at 60 °C and mixed for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by elution (PE / EtOAc = 0-5%) to give product 13-9 (180 mg, yield: 50.3%).

[0835] LCMS: rt = 0.945 min, [M+H] + =637, Purity: 100%

[0836] (10) Resolution of compound 13-9 (yielding 13-9-P1 and 13-9-P2)

[0837]

[0838] The 13-9 sample (180 mg, 0.27 mmol) was further purified by SFC to obtain 13-9-P1 (60 mg, SFC rt = 2.176 min, yield: 67%) and 13-9-P2 (60 mg, SFC rt = 2.68 min, yield: 67%).

[0839] (11) Preparation of compounds 13-A and 13-B

[0840]

[0841] LiOH (20 mg, 0.83 mmol) was added to a solution of 13-9 after resolution (60 mg, 0.09 mmol) in THF / H2O (5 mL), and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product and further purified by preparative HPLC to give 2-(((7aS,11aS)-2-((4-(difluoromethyl)-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid, compound 13-A(1 2.1 mg (yield: 21.2%), and 2-(((7aR,11aR)-2-((4-(difluoromethyl)-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid, for compound 13-B (6.6 mg, yield: 11.6%).

[0842] LCMS: rt = 1.27 min, [M+H] + =623, Purity: 100%

[0843] Example 14

[0844] 2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptadiene[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-((1-(fluoromethyl)cyclopropyl)methyl)-1H-benzo[d]imidazol-6-carboxylic acid (compound 14)

[0845]

[0846] (1) Preparation of compound 14-7

[0847]

[0848] 13-6 (190 mg, 594 mmol) was added to 10 mL of DMF under nitrogen atmosphere and stirred at 25 °C. 4-(bromomethyl)-3-fluorobenzyl nitrile (127.1 mg, 0.594 mmol) and NaH (35.60 mg, 0.89 mmol) were added for 0.5 h. The reaction was monitored by LCMS. The reaction mixture was poured into water (30 mL), extracted with DCM (20 mL × 3), washed with brine, dried, and concentrated to give the crude product. This crude product was further purified by elution (EtOAc / PE = 0-47%) to give 2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptadiene[4,3-b:6,5-c']bipyridine-9(7H)-carboxylate 14-7 (260 mg, yield: 70%).

[0849] LCMS: rt = 2.109 min, [M+H] + =454, Purity: 98%

[0850] (2) Preparation of compound 14-8

[0851]

[0852] A solution of 14-7 (260 mg, 0.57 mmol) in 6 mL HCl / EA was stirred at room temperature for 30 minutes. The reaction was monitored by LCMS. The reaction mixture was concentrated to give crude product 3-fluoro-4-(((5,7,7a,8,9,10,11,11a-octahydrooxetrazine-heptadienyl[4,3-b:6,5-c']dipyridin-2-yl)oxy)methyl)benzyl nitrile 14-8 (200 mg), which was used directly in the next step without purification.

[0853] LCMS: rt = 0.88 min, [M+H] + =355, Purity: 88%

[0854] (3) Preparation of compound 14-9

[0855]

[0856] Under nitrogen atmosphere, at room temperature and with stirring, DIEA (736.7 mg, 0.57 mmol) was added to a solution of 14-8 (200 mg, 0.57 mmol) in 15 mL of MeCN over 10 minutes. Then, 1-int-3 (166.6 mg, 5.7 mmol) was added to the reaction mixture at 60 °C over 16 hours. The reaction was monitored by LC-MS. The reaction mixture was concentrated to a crude product, which was further purified by elution (EtOAc / PE = 0-5%) to give 2-((2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptadiene[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((1-(fluoromethyl)cyclopropyl)methyl)-1H-benzo[d]imidazolium-6-carboxylate 14-9 (200 mg, yield: 57.8%).

[0857] LCMS: rt = 1.553 min, [M+H] + =627, Purity: 98%

[0858] (4) Resolution of compounds 14-9

[0859]

[0860] 14-9 was further purified by SFC method to obtain 14-9-P1 (80 mg, SFC rt = 3.376 min, yield: 88%) and 14-9-P2 (80 mg, SFC rt = 3.926 min, yield: 88%).

[0861] (5) Preparation of compounds 14-A and 14-B

[0862]

[0863] LiOH (15.7 mg, 0.655 mmol) was added to a solution of methyl 2-(((7aS,11aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptatrien[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((1-(fluoromethyl)cyclopropyl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (80 mg, 0.13 mmol) in THF / H2O (5 mL), and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by preparative HPLC to give 2-(((7aS,11aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptadiene[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((1-(fluoromethyl)cyclopropyl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (26.7 mg, yield: 35%).

[0864] LCMS: rt = 1.272 min, [M+H] + =614.2, Purity: 93.7%

[0865] LiOH (15.7 mg, 0.655 mmol) was added to a solution of methyl 2-(((7aR,11aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptatrien[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((1-(fluoromethyl)cyclopropyl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (80 mg, 0.13 mmol) in THF / H2O (5 mL), and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by preparative HPLC to give 2-(((7aR,11aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((1-(fluoromethyl)cyclopropyl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (26.5 mg, yield: 34.7%).

[0866] LCMS: rt = 1.272 min, [M+H] + =614.2, Purity: 98.6%

[0867] Example 15

[0868] 2-(((7aS,11aS)-2-((2-fluoro-4-(trifluoromethyl)benzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid and 2-((( 7aR,11aR)-2-((2-fluoro-4-(trifluoromethyl)benzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (compounds 15A & 15B)

[0869]

[0870] (1) Preparation of compound 15-7

[0871]

[0872] 13-6 (190 mg, 594 mmol) was added to 10 mL of DMF under nitrogen atmosphere and stirred at 25 °C. 1-(bromomethyl)-2-fluoro-4-(trifluoromethyl)benzene (127.1 mg, 0.594 mmol) and NaH (35.60 mg, 0.89 mmol) were added and mixed for 0.5 h. The reaction was monitored by LCMS. The reaction mixture was poured into water (30 mL), extracted with DCM (20 mL × 3), washed with brine, dried, and concentrated to give the crude product. This crude product was further purified by elution (PE / EtOAc = 0-47%) to give 15-7 (260 mg, yield: 70%).

[0873] LCMS: rt = 2.442 min, [M+H] + =441, Purity: 99.7%

[0874] (2) Preparation of compound 15-8

[0875]

[0876] A solution of 15-7 (260 mg, 0.57 mmol) in 6 mL of HCl / EA was stirred at room temperature for 30 minutes. The reaction was monitored by LCMS. The reaction mixture was concentrated to give crude product 15-8 (200 mg), which was used directly in the next step without purification.

[0877] LCMS: rt = 0.88 min, [M+H] +=396, Purity: 98%

[0878] (3) Preparation of compound 15-9

[0879]

[0880] Under nitrogen atmosphere, DIEA (736.7 mg, 0.57 mmol) was added to 15 mL of 15-8 (200 mg, 0.57 mmol) in MeCN, and the mixture was stirred at room temperature for 10 minutes. Then, 1-int-2 (166.6 mg, 5.7 mmol) was added to the reaction mixture at 60 °C for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by elution (PE / EtOAc = 0-5%) to give 15-9 (180 mg, yield: 50.3%).

[0881] LCMS: rt = 0.968 min, [M+H] + =655, Purity: 98.5%

[0882] (4) Resolution of compounds 15-9

[0883]

[0884] 15-9 was further purified by SFC (180 mg, 0.27 mmol) to give 2-(((7aS,11aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate. (60 mg, SFCr. t = 2.176 min, yield 67%) and 2-(((7aR,11aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (60 mg, SFC rt = 2.68 min, yield 67%).

[0885] (5) Preparation of compounds 15-A and 15-B

[0886]

[0887] LiOH (22 mg, 0.92 mmol) was added to a solution of 2-(((7aS,11aS)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (60 mg, 0.09 mmol) in THF / H2O (5 mL). The mixture was stirred at room temperature for 16 hours, and the reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by preparative HPLC to give 2-(((7aS,11aS)-2-((2-fluoro-4-(trifluoromethyl)benzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (42.3 mg, yield: 72.93%).

[0888] LCMS: rt = 1.383 min, [M+H] + =641.2, Purity: 95.6%

[0889] LiOH (22 mg, 0.9 mmol) was added to a solution of 2-(((7aR,11aR)-2-((4-cyano-2-fluorobenzyl)oxy)-5,7a,8,10,11,11a-hexahydrooxetaneheptan[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-1-((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylate (60 mg, 0.09 mmol) in THF / H2O (5 mL), and the mixture was stirred at room temperature for 16 hours. The reaction was monitored by LCMS. The reaction mixture was concentrated to a crude product, which was further purified by preparative HPLC to give 2-(((7aR,11aR)-2-((2-fluoro-4-(trifluoromethyl)benzyl)oxy)-5,7a,8,10,11,11a-hexahydroxyoxetane-heptanetrien[4,3-b:6,5-c']bipyridin-9(7H)-yl)methyl)-1-(((S)-oxetane-2-yl)methyl)-1H-benzo[d]imidazolium-6-carboxylic acid (44.87 mg, yield: 75.6%).

[0890] LCMS: rt = 1.31 min, [M+H] + =641.2, Purity: 100%

[0891] Example 16

[0892] (S)-2-((2-((4-cyano-2-fluorobenzyl)oxy)-5,8,10,11-tetrahydrooxetrazolium hepteno[4,3-b:6,5-c']dipyridin-9(7H)-yl)methyl)-3-(oxetane-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylic acid (compound 16)

[0893]

[0894] (1) Preparation of compound 16-7

[0895]

[0896] Int-5 (0.3 g, 1.2 mmol) was dissolved in anhydrous DMF (5 mL), and then NaH (45 mg, 1.44 mmol) was added in portions at 0 °C. After 5 minutes, a solution of 4-(bromomethyl)-3-fluorobenzyl nitrile (0.202 g, 1.2 mmol) in DMF (5 mL) was added to the reaction mixture through a sleeve. The reaction was monitored by LC-MS after 20 minutes. The reaction mixture was quenched by adding water. The aqueous phase was extracted with EtOAc (20 mL × 3) and washed with brine. The combined organic layers were dried over Na₂SO₄. The product was concentrated and purified by elution (PE / EA = 0-20%) to give product 16-7 (0.35 g, 83.3% yield).

[0897] (2) Preparation of compound 16-8

[0898]

[0899] 16-7 (0.35 g, 0.78 mmol) was added to HCl / EA (20 mL, 3 M). The mixture was stirred at room temperature for 0.5 h. The reaction was detected by LCMS. The mixture was concentrated to give product 16-8 (0.27 g, 98% yield).

[0900] LCMS: rt = 1.31 min, [M+H] + =395, Purity: 95%

[0901] (3) Preparation of compound 16-9

[0902]

[0903] The reaction mixture of 16-8 (96 mg, 0.27 mmol), 1-int-2 (80 mg, 0.27 mmol), and N,N-diisopropylethylamine (175 mg, 1.35 mmol) in 5 mL of acetonitrile was stirred at 60 °C for 16 hours. The reaction mixture was evaporated to dryness, and the residue was purified by SGC (EA / MeOH = 10:1) to give compound 16-9 (100 mg, 60%).

[0904] LCMS: rt = 0.773 min, [M+H] + =653, Purity: 93%

[0905] (4) Preparation of compound 16

[0906]

[0907] At 25 °C, 1 mL of a lithium hydroxide (20 mg, 0.81 mmol) H₂O solution was added to 5 mL of a tetrahydrofuran reaction mixture containing 16-9 (100 mg, 0.16 mmol). The mixture was stirred at 25 °C for 3 hours. The reaction was monitored by LCMS. The reaction mixture was purified by pre-HPLC to give compound 9 (22.45 mg, 22%).

[0908] LCMS: rt = 1.211 min, [M+H] + =640, Purity: 99%

[0909] The GLP-1 receptor agonist and its applications provided by this invention have been described in detail above. Specific embodiments have been used to illustrate the principles and implementation methods of this invention. These embodiments are merely illustrative and are intended to help understand the method and central ideas of this invention. It should be noted that those skilled in the art can make various improvements and modifications to this invention without departing from its principles, and these improvements and modifications also fall under the protection of the claims of this invention.

Claims

1. A compound having the following structure: (I-3-A), Or its pharmaceutically acceptable salt, wherein ------ Indicates whether the key exists or not; Y1 is N; Y2 is selected from CH, N, or C; Y3 is N; R1 is independently selected from -F, -Cl, -CN, -OCH3, -OCH2CH3, -CH3, -CH2CH3, -COCH3, -CONH2, -CF3, -CHF2, -CH2F, -CH2CH2F, -CO-cyclopropyl; R2 is R7-R2', where R7 is -C 1~3 Alkylene, R2' is selected from: , , , , , ; R5 is independently selected from hydrogen, halogen, hydroxyl, -CN, and -C. 1~3 Alkyl, -C 1~3 Alkoxy, -C 3~6 cycloalkyl; R6 is selected from -COOH, -CH2COOH, -CH2CH2COOH, and -CH(CH3)COOH; n is selected from 1, 2, or 3; p is selected from 0, 1, or 2; R4 and R5 are cyclically transformed into 5- to 8-membered rings, wherein the 5- to 8-membered rings are selected from: , , , , , Furthermore, the resulting 5- to 8-membered rings can be arbitrarily formed from C, provided that the oxidation state allows. 1-3 Alkyl, C 1-3 Halogenated alkyl, halogen, cyano, oxo, C 1-3 Alkoxy substitution 1 to 3 times.

2. The compound according to claim 1, characterized in that, Any adjacent R4 and R5 rings are transformed into 5- to 8-membered rings, wherein the 5- to 8-membered rings are selected from: , , , , .

3. The compound according to claim 1, characterized in that, Any adjacent R4 and R5 rings are transformed into 5- to 8-membered rings, wherein the 5- to 8-membered rings are selected from: , .

4. The compound according to claim 1, characterized in that, The n=2 mentioned above.

5. The compound according to claim 1, characterized in that, The p=1 mentioned above.

6. The compound according to claim 1, characterized in that, The R6 mentioned is -COOH.

7. The compound according to claim 1, wherein, R7 is a -C1 alkylene group, and R2' is selected from: , .

8. A compound, characterized in that, The compounds mentioned are selected from: , , , , , , , , , , , , , , , , , , , , , , Or, or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition comprising the compound of any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable pharmaceutical carrier.

10. The use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8 in the preparation of a medicament for treating GLP-1 receptor agonist-mediated diseases or related diseases, wherein the GLP-1-mediated diseases and related diseases are selected from: diabetes mellitus, hyperglycemia, insulin resistance, glucose intolerance, diabetic nephropathy, diabetic neuropathy, diabetic retinopathy, adipocyte dysfunction, obesity, dyslipidemia, and hyperinsulinemia.

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