Non-fluoroquinolone compounds and uses thereof

Abstract

The application discloses a new non-fluorine quinolone compound and application thereof, and particularly relates to a compound shown in formula (II) and pharmaceutically acceptable salts thereof.

Description

[0001] This application claims the following priority:

[0002] CN202111444560.2, November 30, 2021;

[0003] CN202210864782.8, July 21, 2022. Technical Field

[0004] This invention relates to a novel class of nonfluoroquinolone compounds and their applications, specifically to compounds represented by formula (II) and their pharmaceutically acceptable salts. Background Technology

[0005] Quinolones were introduced in the 1960s and have undergone continuous updates and improvements over the decades, leading to their widespread clinical application. Due to their broad antibacterial spectrum, strong antibacterial activity, convenient administration, and lack of cross-resistance with other commonly used antibiotics, quinolones have rapidly gained popularity in clinical use, becoming one of the most commonly used and important drugs worldwide. With ongoing modifications and research into these drugs, many other biological activities have been discovered, including reported antitumor, antiviral, and anti-inflammatory effects.

[0006] Acne affects more than 85% of human skin conditions. It commonly occurs on the face, neck, and upper trunk. Although acne is caused by multiple factors, symbiotic skin bacteria (Propionibacterium acnes) play a major role in the formation of acne lesions. It is an infection of the pilosebaceous sebaceous glands and oil glands in the skin. Propionibacterium acnes can also activate Toll-like receptor 2 (TLR2) on innate immune cells. TLR activation can trigger the expression of various cytokines (such as IL-6, IL-8, and IL-12) and stimulate the recruitment of chemokines by other host immune cells.

[0007] Erythromycin and clindamycin are the most widely used antibiotics for treating acne. However, due to their long-term and widespread use, drug resistance is becoming increasingly serious, necessitating the development of new antibiotics for acne treatment. In recent years, numerous patents and articles have reported that quinolone drugs possess good anti-Propionibacterium acnes activity, while also exhibiting anti-inflammatory activity. Therefore, researching next-generation quinolone drugs for acne treatment has significant theoretical and economic value. Summary of the Invention

[0008] This invention provides compounds of formula (II), stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0009]

[0010] in,

[0011] Selected from single and double bonds;

[0012] T1 is selected from N, NH, CRI, and N. + R1(A - );

[0013] T2 is selected from N and CR2;

[0014] A - Selected from F - Cl - ,Br - I - OH - and HCO3 - ;

[0015] X is selected from -C(R7R8)- and -C(R7R8)-C(R7R8)-;

[0016] Y is selected from -O-, -S-, -NH-, and -C(R7R8)-;

[0017] Z is selected from -C(R7R8)-;

[0018] Alternatively, -YZ- is selected from -C(R9)=C(R9)-;

[0019] R1 is selected from H, F, Cl, Br, I, -OH, -NH2, -CN, and -CH3;

[0020] R2 is selected from H, F, Cl, Br, I, -OH, -NH2, -CN, and C. 1-3 Alkyl, wherein the C 1-3 Alkyl groups may be optionally surrounded by 1, 2, or 3 R's. a replace;

[0021] R3 is selected from H, F, Cl, Br, I, =O, -OH, -NH2, -CN, -NHC(=NH)NH2, C 1-3 Alkyl, C 1-3 Alkylamino and C 1-3 Alkoxy, wherein the C 1-3 Alkyl, C 1-3 Alkylamino and C 1-3 The alkoxy groups are independently and optionally surrounded by 1, 2, or 3 R groups. b replace;

[0022] Alternatively, R1, together with R3 and the atoms attached to them, forms a 5-6 membered heterocyclic alkyl, 5-6 membered heterocyclic alkenyl, or 5-6 membered heteroaryl, wherein the 5-6 membered heterocyclic alkyl, 5-6 membered heterocyclic alkenyl, and 5-6 membered heteroaryl are each independently and optionally separated by 1, 2, 3, or 4 R atoms. c replace;

[0023] R4 is selected from H and C. 1-3 Alkyl, C 3-6 Cycloalkyl, phenyl, and 5-6 heteroaryl groups, wherein the C 1-3 Alkyl, C 3-6 The cycloalkyl, phenyl, and 5-6 heteroaryl groups are each independently and optionally bound by 1, 2, 3, or 4 R groups. d replace;

[0024] R5 is selected from H, F, Cl, Br, I, -OH, -NH2, and -CN;

[0025] R6 is selected from H, F, Cl, Br, I, -OH, -NH2, and -CN;

[0026] Each of R7, R8, and R9 is independently selected from H, F, Cl, Br, I, -OH, -NH2, and -CN, respectively;

[0027] Each R a Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0028] Each R b Each element is independently selected from F, Cl, Br, I, -OH, -NH2, -CN, and C. 1-3 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, 5-6 membered heteroaryl and phenyl, wherein C 1-3 Alkoxy, C 3-6 Cycloalkyl, 3-6-membered heterocycloalkyl, 5-6-membered heteroaryl and phenyl are each independently and optionally substituted with 1, 2, 3 or 4 Rs;

[0029] Each R c Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, -CN, and -CH3;

[0030] Each R d Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0031] Each R is independently selected from F, Cl, Br, I, -OH, -NH2, -NO2, -CN, -ONHC(=NH)NH2 and C. 1-3 alkyl;

[0032] In the 5-6 membered heterocyclic alkyl, 3-6 membered heterocyclic alkyl, 5-6 membered heterocyclic alkenyl and 5-6 membered heteroaryl, "hetero" means 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from O, NH, S and N.

[0033] This invention provides compounds of formula (II), stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0034]

[0035] in,

[0036] Selected from single and double bonds;

[0037] T1 is selected from N and CR1;

[0038] T2 is selected from N and CR2;

[0039] X is selected from -C(R7R8)- and -C(R7R8)-C(R7R8)-;

[0040] Y is selected from -O-, -S-, -NH-, and -C(R7R8)-;

[0041] Z is selected from -C(R7R8)-;

[0042] Alternatively, -YZ- is selected from -C(R9)=C(R9)-;

[0043] R1 is selected from H, F, Cl, Br, I, -OH, -NH2, and -CN;

[0044] R2 is selected from H, F, Cl, Br, I, -OH, -NH2, -CN, and C. 1-3 Alkyl, wherein the C 1-3 Alkyl groups may be optionally surrounded by 1, 2, or 3 R's. a replace;

[0045] R3 is selected from H, F, Cl, Br, I, =O, -OH, -NH2, -CN, C 1-3 Alkyl, C 1-3 Alkylamino and C 1-3 Alkoxy, wherein the C 1-3 Alkyl, C 1-3 Alkylamino and C 1-3 The alkoxy groups are independently and optionally surrounded by 1, 2, or 3 R groups. b replace;

[0046] Alternatively, R1 and R3, together with the carbon atoms attached to them, form 5-6 membered heterocyclic alkyl, 5-6 membered heterocyclic alkenyl, and 5-6 membered heteroaryl groups, wherein the 5-6 membered heterocyclic alkyl, 5-6 membered heterocyclic alkenyl, and 5-6 membered heteroaryl groups are each independently and optionally separated by 1, 2, 3, or 4 R atoms. c replace;

[0047] R4 is selected from H and C. 1-3 Alkyl, C 3-6 Cycloalkyl, phenyl, and 5-6 heteroaryl groups, wherein the C 1-3 Alkyl, C 3-6The cycloalkyl, phenyl, and 5-6 heteroaryl groups are each independently and optionally bound by 1, 2, 3, or 4 R groups. d replace;

[0048] R5 is selected from H, F, Cl, Br, I, -OH, -NH2, and -CN;

[0049] R6 is selected from H, F, Cl, Br, I, -OH, -NH2, and -CN;

[0050] Each of R7, R8, and R9 is independently selected from H, F, Cl, Br, I, -OH, -NH2, and -CN, respectively;

[0051] R a Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0052] R b Each element is independently selected from F, Cl, Br, I, -OH, -NH2, -CN, and C. 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl and phenyl, wherein the C 3-6 Cycloalkyl, 3-6 membered heterocyclic alkyl and phenyl groups are each independently and optionally substituted with 1, 2, 3 or 4 R groups;

[0053] R c Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0054] R d Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0055] R is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0056] In the 5-6 membered heterocyclic alkyl, 3-6 membered heterocyclic alkyl, 5-6 membered heterocyclic alkenyl and 5-6 membered heteroaryl, "hetero" means 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -O-, -NH-, -S- and -N-.

[0057] This invention provides compounds of formula (I), stereoisomers thereof, or pharmaceutically acceptable salts thereof.

[0058]

[0059] in,

[0060] T1 is selected from N and CR1;

[0061] T2 is selected from N and CR2;

[0062] X is selected from -C(R7R8)- and -C(R7R8)-C(R7R8)-;

[0063] Y is selected from -O- and -C(R7R8)-;

[0064] Z is selected from -C(R7R8)-;

[0065] Alternatively, -YZ- is selected from -C(R9)=C(R9)-;

[0066] R1 is selected from H, F, Cl, Br, I, -OH, -NH2, and -CN;

[0067] R2 is selected from H, F, Cl, Br, I, -OH, -NH2, -CN, and C. 1-3 Alkyl, wherein the C 1-3 Alkyl groups may be optionally surrounded by 1, 2, or 3 R's. a replace;

[0068] R3 is selected from H, F, Cl, Br, I, -OH, -NH2, -CN, C 1-3 Alkylamino and C 1-3 Alkoxy, wherein the C 1-3 Alkylamino and C 1-3 The alkoxy groups are independently and optionally surrounded by 1, 2, or 3 R groups. b replace;

[0069] Alternatively, R1 and R3 together with the carbon atoms attached to them form 5-6 membered heterocyclic alkyl, 5-6 membered heterocyclic alkenyl, and 5-6 membered heteroaryl groups, wherein the 5-6 membered heterocyclic alkenyl and 5-6 membered heteroaryl groups are each independently and optionally surrounded by 1, 2, 3, or 4 R atoms. c replace;

[0070] R4 is selected from H and C. 1-3 Alkyl, C 3-6 Cycloalkyl, phenyl, and 5-6 heteroaryl groups, wherein the C 1-3 Alkyl, C 3-6 The cycloalkyl, phenyl, and 5-6 heteroaryl groups are each independently and optionally bound by 1, 2, 3, or 4 R groups. d replace;

[0071] R5 is selected from H, F, Cl, Br, I, -OH, -NH2, and -CN;

[0072] R6 is selected from H, F, Cl, Br, I, -OH, -NH2, and -CN;

[0073] Each of R7, R8, and R9 is independently selected from H, F, Cl, Br, I, -OH, -NH2, and -CN, respectively;

[0074] R aEach of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0075] R b Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0076] R c Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0077] R d Each of the following is independently selected from F, Cl, Br, I, -OH, -NH2, and -CN;

[0078] In the 5-6 membered heterocyclic alkyl, 5-6 membered heterocyclic alkenyl and 5-6 membered heteroaryl, "hetero" means 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -O-, -NH-, -S- and -N-.

[0079] In some embodiments of the present invention, the above-mentioned Selected from double bonds, other variables are as defined in this invention.

[0080] In some embodiments of the present invention, T1 is selected from N, and other variables are as defined in the present invention.

[0081] In some embodiments of the present invention, T1 is selected from CH, and other variables are as defined in the present invention.

[0082] In some embodiments of the present invention, T1 is selected from NH, and other variables are as defined in the present invention.

[0083] In some embodiments of the present invention, T1 is selected from N. + R1(A - Other variables are as defined in this invention.

[0084] In some embodiments of the present invention, T1 is selected from N. + R1(Cl - Other variables are as defined in this invention.

[0085] In some embodiments of the present invention, T2 is selected from N, and other variables are as defined in the present invention.

[0086] In some embodiments of the present invention, T2 is selected from CH, and other variables are as defined in the present invention.

[0087] In some embodiments of the present invention, the above-mentioned A - Selected from F - Cl - ,Br - and I - Other variables are as defined in this invention.

[0088] In some embodiments of the present invention, the above-mentioned A - Selected from Cl - Other variables are as defined in this invention.

[0089] In some embodiments of the present invention, X is selected from -CH2-, and other variables are as defined in the present invention.

[0090] In some embodiments of the present invention, Y is selected from -O-, and other variables are as defined in the present invention.

[0091] In some embodiments of the present invention, Y is selected from -CH2-, and other variables are as defined in the present invention.

[0092] In some embodiments of the present invention, Z is selected from -CH2-, and other variables are as defined in the present invention.

[0093] In some embodiments of the present invention, the above-mentioned -YZ- is selected from -CH=CH-, and other variables are as defined in the present invention.

[0094] In some embodiments of the present invention, each of the above R is independently selected from -NO2, -ONHC(=NH)NH2 and -CH3, and other variables are as defined in the present invention.

[0095] In some embodiments of the present invention, the above-mentioned R b The radicals are independently selected from F, Cl, Br, -OH, -NH2, -OCH2CH3, cyclobutyl, oxetyl, oxetyl, azirbutyl, 5-membered heteroaryl, and phenyl, wherein -OCH2CH3, cyclobutyl, oxetyl, oxetyl, azirbutyl, 5-membered heteroaryl, and phenyl are independently and optionally substituted by 1, 2, 3, or 4 Rs, and R and other variables are as defined in this invention.

[0096] In some embodiments of the present invention, the above-mentioned R b The components are independently selected from F, Cl, Br, -OH, -NH2, cyclobutyl, oxetyl, azirbutyl, and phenyl, wherein the cyclobutyl, oxetyl, azirbutyl, and phenyl are independently and optionally substituted by 1, 2, 3, or 4 Rs, and R and other variables are as defined in this invention.

[0097] In some embodiments of the present invention, the above-mentioned R b The radicals are independently selected from F, Cl, Br, -OH, -NH2, -OCH2CH3, cyclobutyl, oxobutyl, 1,3-dioxopentane, acridine, imidazole, and phenyl, respectively, wherein -OCH2CH3, cyclobutyl, oxobutyl, 1,3-dioxopentane, acridine, imidazole, and phenyl are each independently and optionally substituted by 1, 2, 3, or 4 Rs, and R and other variables are as defined in this invention.

[0098] In some embodiments of the present invention, the above-mentioned R b Each of the following is independently selected from F, Cl, Br, -OH, -NH2, -OCH2CH3, The -OCH2CH3, Each R can be independently and arbitrarily replaced by 1, 2, 3 or 4 Rs, and R and other variables are as defined in this invention.

[0099] In some embodiments of the present invention, the above-mentioned R b Each is independently selected from -OH, -NH2, The above Each R can be independently and arbitrarily replaced by 1, 2, 3 or 4 Rs, and R and other variables are as defined in this invention.

[0100] In some embodiments of the present invention, the above-mentioned R b Each is independently selected from F, -OH, -NH2, Other variables are as defined in this invention.

[0101] In some embodiments of the present invention, the above-mentioned R b Each is independently selected from -OH, -NH2, Other variables are as defined in this invention.

[0102] In some embodiments of the present invention, the above-mentioned R b The variables are independently selected from F, Cl, Br and -OH, respectively, and other variables are as defined in this invention.

[0103] In some embodiments of the present invention, the above-mentioned R c Each variable is independently selected from -CH3, and other variables are as defined in this invention.

[0104] In some embodiments of the present invention, the above-mentioned R d The variables are independently selected from F, Cl, Br and -NH2, respectively, and other variables are as defined in this invention.

[0105] In some embodiments of the present invention, the above-mentioned R d The variables are independently selected from F and -NH2, respectively, and other variables are as defined in this invention.

[0106] In some embodiments of the present invention, R1 is selected from H, and other variables are as defined in the present invention.

[0107] In some embodiments of the present invention, R2 is selected from H and -CH3, and other variables are as defined in the present invention.

[0108] In some embodiments of the present invention, R2 is selected from H, and other variables are as defined in the present invention.

[0109] In some embodiments of the present invention, the aforementioned R3 is selected from H, F, Cl, Br, =O, -OH, -NH2, -CN, -NHC(=NH)NH2, -CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, -NHCH2CH2CH3, -N(CH3)CH2CH3, and -OCH3, wherein -NHC(=NH)NH2, -CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, -NHCH2CH2CH3, -N(CH3)CH2CH3, and -OCH3 are each independently and optionally selected by 1, 2, or 3 R3. b Replace, R b Other variables are as defined in this invention.

[0110] In some embodiments of the present invention, the R3 is selected from H, F, Cl, Br, =O, -OH, -NH2, -CN, -CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, -NHCH2CH2CH3, and -OCH3, wherein -CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, -NHCH2CH2CH3, and -OCH3 are each independently and optionally selected by 1, 2, or 3 R3. b Replace, R b Other variables are as defined in this invention.

[0111] In some embodiments of the present invention, the R3 is selected from H, F, Cl, Br, -OH, -NH2, -CN, -NHCH3, -N(CH3)2, -NCH2CH3, and -OCH3, wherein NHCH3, -N(CH3)2, -NCH2CH3, and -OCH3 are each independently and optionally selected by 1, 2, or 3 R3. b Replace, R b Other variables are as defined in this invention.

[0112] In some embodiments of the present invention, the R3 is selected from H, =O, -NH2, -NHC(=NH)NH2, -CH3, -NHCH3, -N(CH3)2, -NCH2CH3, -NHCH2CH2CH3, -NHCH2CH2OH, -NHCH2CH2NH2, -OCH3, Other variables are as defined in this invention.

[0113] In some embodiments of the present invention, R3 is selected from H, =O, -NH2, -CH3, -NHCH3, -N(CH3)2, -NCH2CH3, -NHCH2CH2CH3, -NHCH2CH2OH, -NHCH2CH2NH2, -OCH3, Other variables are as defined in this invention.

[0114] In some embodiments of the present invention, R3 is selected from -NH2, -NHCH3, -N(CH3)2, -NCH2CH3, -NCH2CH2OH and -OCH3, and other variables are as defined in the present invention.

[0115] In some embodiments of the present invention, R4 is selected from C. 3-6 cycloalkyl, wherein the C 3-6 The cycloalkyl group is optionally surrounded by 1, 2, 3 or 4 R's. d Replacement, other variables as defined in this invention.

[0116] In some embodiments of the present invention, R4 is selected from H, -CH2CH3, cyclopropyl, phenyl, and pyridyl, wherein -CH2CH3, cyclopropyl, phenyl, and pyridyl are each independently and optionally converted by 1, 2, 3, or 4 Rs. d Replacement, other variables as defined in this invention.

[0117] In some embodiments of the present invention, R4 is selected from H, -CH2CH2F, Other variables are as defined in this invention.

[0118] In some embodiments of the present invention, R4 is selected from... Other variables are as defined in this invention.

[0119] In some embodiments of the present invention, R5 and R6 are independently selected from H and F, respectively, and other variables are as defined in the present invention.

[0120] In some embodiments of the present invention, R6 is selected from H and F, and other variables are as defined in the present invention.

[0121] In some embodiments of the present invention, R5 and R6 are each independently selected from H, and other variables are as defined in the present invention.

[0122] In some embodiments of the present invention, each of R7, R8 and R9 is independently selected from H, and other variables are as defined in the present invention.

[0123] In some embodiments of the present invention, R1 and R3, along with the atoms connected to them, are formed together. Quaternary ammonium salt ring, wherein Each can be independently selected by 1, 2, 3 or 4 Rs.c Replace, R c Other variables are as defined in this invention.

[0124] In some embodiments of the present invention, R1 and R3, along with the atoms connected to them, are formed together. Quaternary ammonium salt ring, other variables as defined in this invention.

[0125] In some embodiments of the present invention, R1 and R3, along with the atoms connected to them, are formed together. Quaternary ammonium salt ring, wherein Each can be independently selected by 1, 2, 3 or 4 Rs. c Replace, R c Other variables are as defined in this invention.

[0126] In some embodiments of the present invention, R1 and R3, along with the atoms connected to them, are formed together. Quaternary ammonium salt ring, other variables as defined in this invention.

[0127] In some embodiments of the present invention, R1 and R3, along with the atoms connected to them, are formed together. The above Each can be independently selected by 1, 2, 3 or 4 Rs. c Replace, R c Other variables are as defined in this invention.

[0128] In some embodiments of the present invention, R1 and R3, along with the carbon atoms bonded to them, are formed together. Among them, the Each can be independently selected by 1, 2, 3 or 4 Rs. c Replace, R c Other variables are as defined in this invention.

[0129] In some embodiments of the present invention, R1 and R3, along with the atoms connected to them, are formed together. Other variables are as defined in this invention.

[0130] In some embodiments of the present invention, R1 and R3, along with the carbon atoms bonded to them, are formed together. Other variables are as defined in this invention.

[0131] In some embodiments of the present invention, the above-mentioned compound has the structure shown in formula (Ⅱ-1):

[0132]

[0133] in, X, Y, Z, T1, T2, R3, R5 and R6 are as defined in this invention.

[0134] In some embodiments of the present invention, the above-mentioned compound has the structure shown in formula (Ⅱ-2):

[0135]

[0136] in, Y, Z, T1, T2, and R3 are as defined in this invention.

[0137] In some embodiments of the present invention, the above-mentioned compounds have the structures shown in formulas (II-3), (II-4), (II-5), and (II-6):

[0138]

[0139]

[0140] Wherein, T1, T2 and R3 are as defined in this invention.

[0141] In some embodiments of the present invention, the above-mentioned compounds have structures shown in formulas (Ⅱ-3A), (Ⅱ-3B), (Ⅱ-3C), (Ⅱ-3D), (Ⅱ-3E), (Ⅱ-5A), and (Ⅱ-6A):

[0142]

[0143] Among them, A - R1, R2 and R3 are as defined in this invention.

[0144] Some solutions in this invention are derived from arbitrary combinations of the above-mentioned variables.

[0145] The present invention also provides compounds of the following formula, stereoisomers, or pharmaceutically acceptable salts thereof.

[0146]

[0147]

[0148]

[0149] The present invention also provides compounds of the following formula, stereoisomers, or pharmaceutically acceptable salts thereof.

[0150]

[0151] This invention also provides the use of the above-mentioned compounds, stereoisomers or pharmaceutically acceptable salts thereof in the preparation of antibacterial and anti-inflammatory drugs.

[0152] In some embodiments of the present invention, the above application is preferably used in the preparation of acne treatment drugs.

[0153] This invention also provides the use of the above-mentioned compounds, stereoisomers or pharmaceutically acceptable salts thereof in the preparation of acne treatment drugs.

[0154] This invention also provides a biological experimental testing method for the above-mentioned compounds:

[0155] Experimental Test Method 1: Propionibacterium acnes-induced THP-1 cell inflammation model experiment

[0156] Experimental objective:

[0157] Establish a Propionibacterium acnes-induced inflammation model of THP-1 cells.

[0158] Laboratory consumables:

[0159] Strains: Propionibacterium acnes ATCC 6919; Culture medium: Brucella agar (containing 5% LHB + 5μg / ml Hemin + 1μg / ml Vitamin K1); Cells: THP-1 ATCC TIB-202; Culture medium: RPMI 1640 (gibco 22400-089) + 10% heat-inactivated FBS + 1×GlutaMAX (100×GlutaMAX:gibco 3050-061).

[0160] Experimental plan:

[0161] a) Preparation of inflammatory stimulants

[0162] i. Inoculate Propionibacterium acnes ATCC 6919 glycerol tubes into Brucella agar (containing 5% LHB + 5 μg / mL Hemin + 1 μg / mL Vitamin K1) and anaerobically culture at 37°C for 48 h.

[0163] ii. Collect the plate culture and suspend it in PBS, then adjust the bacterial count to 3 × 10⁻⁶. 9 CFU / mL. Then inactivate by placing in a water bath at 80°C for 30 minutes. CFU counts should be performed before and after bacterial inactivation.

[0164] iii. Store the inactivated bacterial suspension in a -80°C refrigerator for later use.

[0165] b) Preparation of THP-1 in cells

[0166] i. Cell resuscitation and culture: THP-1 cells frozen in liquid nitrogen were resuscitated in RPMI 1640 (containing 10% FBS and 1×glutamax). They can be used for experiments after one passage.

[0167] ii. Cell seeding: Seed 2 × 10⁶ cells / year. 5 190 μL, 1.1 × 10⁻⁶ cells (190 μL, 1.1 × 10⁻⁶ cells) 6 (cfu / mL) was poured into a 96-well flat-bottom plate and set aside for use.

[0168] c) Inducing stimulation of cells

[0169] i. 8 μL of heat-inactivated Propionibacterium acnes (tested at two concentrations: 3 × 10⁸) 9 cfu / mL (high concentration), 6×10 8 Cells were stimulated with a low concentration of CFU / mL to induce inflammatory cytokines, and two replicates were performed. After 1 h of induction, the test plate was incubated at 37 °C for 24 h, 48 h, and 72 h. PBS was used as a negative control. After incubation, the test plate was centrifuged (4000 RPM, 5 min). The supernatant was collected and stored at -80 °C for later use.

[0170] d) Enzyme-linked immunosorbent assay (ELISA)

[0171] i. Detect the cytokine IL-8 level in the cell supernatant according to the instructions of the Human CXCL8 / IL-8 Quantikine ELISA Kit (R&D-D8000C).

[0172] ii. Detect the cytokine IL-6 level in the cell supernatant according to the Human IL-6 Quantikine ELISA Kit (R&D-D6050) instructions.

[0173] Technical effect

[0174] This invention synthesizes the compound of formula (I) and its pharmaceutically acceptable salt through a simple preparation method, obtaining a new class of non-fluoroquinolone antibiotics that target both DNA gyrase and TLR-MD2, for the treatment of acne caused by Propionibacterium acnes, thus solving the problem of drug resistance in Propionibacterium acnes.

[0175] Definitions and Explanations

[0176] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0177] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0178] The term "pharmaceutically acceptable salt" refers to the salt of the compounds of this invention, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this invention, with a specific substituent. When the compounds of this invention contain relatively acidic functional groups, a base addition salt can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of this invention contain relatively basic functional groups, an acid addition salt can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds of this invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt. This invention also contemplates the formation of N-cationic quaternary ammonium salts from any compound containing an N-group. + R1(A - ) represents a quaternary ammonium salt where the heteroatom N is a positive ion, in which case A - For F - Cl - ,Br - I - OH - HCO3 - HSO3 - HSO4 - NO3 - ,ClO - BrO - I3 - ClO3 - ClO4 - HCOO - CH3COO - H2PO4 - wait.

[0179] 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.

[0180] In addition to the salt form, the compounds provided by this invention also exist in prodrug form. The prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to be converted into the compounds of this invention. Furthermore, the prodrugs can be converted into the compounds of this invention in the in vivo environment via chemical or biochemical methods.

[0181] Some compounds of this invention may exist in non-solventized or solvated forms, including hydrated forms. Generally, solvated and non-solventized forms are equivalent and both are included within the scope of this invention.

[0182] Unless otherwise stated, the term "treatment" is intended to refer to all processes that may slow, interrupt, halt, or prevent the progression of a disease, but does not necessarily mean that all symptoms are completely eliminated.

[0183] 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.

[0184] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0185] 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.

[0186] 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.

[0187] Unless otherwise stated, "(+)" indicates right-handed rotation, "(-)" indicates left-handed rotation, and "(±)" indicates racemic rotation.

[0188] 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 The relative configuration of the center of a solid is indicated by a wavy line. 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

[0189] The compounds of this invention can exist in specific forms. Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. If tautomerization is possible (e.g., in solution), chemical equilibrium of the tautomer can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions involving the rearrangement of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between the two tautomers, pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0190] Unless otherwise stated, the terms "rich in one isomer," "isomer enrichment," "rich in one enantiomer," or "enantiomer enrichment" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0191] Unless otherwise stated, the terms "isomer excess" or "enantiomer excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomer excess (ee value) is 80%.

[0192] 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 amino group) 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, optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0193] 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.

[0194] The terms “optional” or “optionally” refer to events or conditions that may occur but are not required to occur as described below, and the description includes both cases where said events or conditions occur and cases where said events or conditions do not occur.

[0195] The term "substituted" refers to the substitution of one or more hydrogen atoms on a specific atom by a substituent. Substituents can include deuterium and hydrogen variants, provided the valence state of the specific atom is normal and the resulting compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are substituted. For example, R3 in this invention is selected when =O, the structural unit... middle Selected from single bonds, and structural units Selected from

[0196] The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents may be arbitrary on the basis of chemical feasibility.

[0197] 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.

[0198] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0199] 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.

[0200] 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.

[0201] 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 pyridinium substituent can be attached to the substituted group by any carbon atom on the pyridine ring.

[0202] 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 combinations produce stable compounds. Unless otherwise specified, when a group has one or more linkable sites, any one or more sites of that group may be connected to other groups by chemical bonds. When the chemical bond connection is non-directional and an H atom is present at the linkable site, the number of H atoms at that site decreases accordingly with the number of chemical bonds connected, resulting in a group with the 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 diagram indicate that the phenyl group is connected to other groups through the carbon atoms at positions 1 and 2. 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.

[0203] Unless otherwise specified, the term "alkyl" on its own or as part of another substituent refers to a straight-chain or branched saturated hydrocarbon group. The alkyl group can be C10 or C20. 1-6 Alkyl or C 1-3 Alkyl group. The alkyl group may optionally be substituted with one or more of the following groups: oxo, hydroxy, amino, nitro, halogen, cyano, alkenyl, alkoxy, haloalkoxy, alkylamino, dialkylamino, haloalkylamino, halodialkylamino, cycloalkyl, cycloalkyloxy, heterocyclic, heterocyclic alkyl, heterocyclic alkyloxy, heterocyclic alkyl, heterocyclic alkyloxy, heteroaryl, heteroaryloxy, aryl or aryloxy.

[0204] Unless otherwise specified, the term "C" 1-3 "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

[0205] Unless otherwise specified, the term "alkoxy group" refers to those alkyl groups that are attached to the remainder of a molecule via an oxygen atom. The alkoxy group can be C10 or C20. 1-6 Alkoxy or C 1-3 Alkoxy group. The alkoxy group may optionally be substituted with one or more of the following groups: oxo, hydroxy, amino, nitro, halogen, cyano, alkenyl, alkoxy, haloalkoxy, alkylamino, dialkylamino, haloalkylamino, halodialkylamino, cycloalkyl, cycloalkyloxy, heterocyclic, heterocyclic alkyloxy, heterocyclic alkyl, heterocyclic alkyloxy, heteroaryl, heteroaryloxy, aryl or aryloxy.

[0206] Unless otherwise specified, the term "C" 1-3 "Alkoxy" refers to alkyl groups containing 1 to 3 carbon atoms that are attached to the rest of the molecule by an oxygen atom. The C1-3 Alkoxy groups include C 1-2 C 2-3 C3 and C2 alkoxy groups, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

[0207] Unless otherwise specified, the term "C" 1-3 "Alkylamino" refers to alkyl groups containing 1 to 3 carbon atoms that are attached to the rest of the molecule via an amino group. The C 1-3 Alkylamino groups include C 1-2 C3 and C2 alkylamino groups, etc. C 1-3 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCH2(CH3)2, etc.

[0208] Unless otherwise specified, the term “halogen” or “halogen” itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.

[0209] The term "cycloalkyl" refers to a fully saturated carbon ring that can exist as a monocyclic, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 12-membered ring. Non-limiting examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, etc. The cycloalkyl group can be C16-26-3 ... 3-6 Cycloalkyl. The cycloalkyl group is optionally substituted with one or more of the following groups: oxo, hydroxy, amino, nitro, halogen, cyano, alkyl, alkenyl, alkoxy, haloalkoxy, alkylamino, dialkylamino, haloalkylamino, halodialkylamino, carboxyl, -C(O)O-alkyl, -OC(O)-alkyl, -C(O)NH2, -C(O)NH-alkyl, -C(O)N(alkyl)2, -NHC(O)-alkyl, -C(O)-alkyl, -S(O)-alkyl, -S(O)2-alkyl, -S(O)2NH2, -S(O)2NH-alkyl, -S(O)2N(alkyl)2, cycloalkyl, cycloalkyloxy, heterocyclic, heterocyclic alkyloxy, heterocyclic alkyl, heterocyclic alkyloxy, heteroaryl, heteroaryloxy, aryl or aryloxy.

[0210] Unless otherwise specified, "C 3-6 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which can be monocyclic or bicyclic. 3-6 Cycloalkyl groups include C 3-5 C 4-5 and C 5-6Cycloalkyl groups, etc.; they can be monovalent, divalent, or polyvalent. C 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0211] The term "heterocyclic group" refers to a non-aromatic ring that is fully saturated or partially unsaturated (but not fully unsaturated) and can exist as a monocyclic, bridged, or spirocyclic ring. Unless otherwise indicated, the heterocycle is typically a 3- to 12-membered ring containing 1 to 3 heteroatoms independently selected from sulfur, oxygen, and / or nitrogen (preferably 1 or 2 heteroatoms). Non-limiting examples of heterocyclic groups include, but are not limited to, ethylene oxide, tetrahydrofuranyl, dihydrofuranyl, pyrrolyl, N-methylpyrrolyl, dihydropyrrolyl, piperidinyl, piperazinyl, pyrazolyl, 4H-pyranyl, morpholinyl, thiomorpholinyl, tetrahydrothiophene, etc. The heterocyclic group is optionally substituted with one or more of the following groups: oxo, hydroxy, amino, nitro, halogen, cyano, alkyl, alkenyl, alkoxy, haloalkoxy, alkylamino, dialkylamino, haloalkylamino, halodialkylamino, carboxyl, -C(O)O-alkyl, -OC(O)-alkyl, -C(O)NH2, -C(O)NH-alkyl, -C(O)N(alkyl)2, -NHC(O)-alkyl, -C(O)-alkyl, -S(O)-alkyl, -S(O)2-alkyl, -S(O)2NH2, -S(O)2NH-alkyl, -S(O)2N(alkyl)2, cycloalkyl, cycloalkyloxy, heterocyclic, heterocyclic alkyloxy, heterocyclic alkyl, heterocyclic alkyloxy, heteroaryl, heteroaryloxy, aryl or aryloxy.

[0212] The term "heterocyclic alkyl" refers to a fully saturated cyclic group that may exist as a monocyclic, bridged, or spirocyclic ring. Unless otherwise indicated, the heterocycle is typically a 4- to 12-membered ring containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms) independently selected from sulfur, oxygen, and / or nitrogen. Non-limiting examples of 4-membered heterocyclic alkyl groups include, but are not limited to, acridinel, oxadiazolyl, and thiobutylyl; examples of 5-membered heterocyclic alkyl groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, imidazolyl, and tetrahydropyrazolyl; examples of 6-membered heterocyclic alkyl groups include, but are not limited to, piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, morpholinyl, piperazine, 1,4-thiaoxalyl, 1,4-dioxane, thiomorpholinyl, 1,3-dithiaalkyl, and 1,4-dithiaalkyl; and examples of 7-membered heterocyclic alkyl groups include, but are not limited to, azaheptanyl, oxaheptanyl, and thioheptanyl. The heterocyclic alkyl group may be a 4- to 6-membered heterocyclic alkyl group, or a monocyclic heterocyclic alkyl group having 5 or 6 ring atoms. The heterocyclic alkyl group is optionally substituted with one or more of the following groups: oxo, hydroxy, amino, nitro, halogen, cyano, alkyl, alkenyl, alkoxy, haloalkoxy, alkylamino, dialkylamino, haloalkylamino, halodialkylamino, carboxyl, -C(O)O-alkyl, -OC(O)-alkyl, -C(O)NH2, -C(O)NH-alkyl, -C(O)N(alkyl)2, -NHC(O)-alkyl, -C(O)-alkyl, -S(O)-alkyl, -S(O)2-alkyl, -S(O)2NH2, -S(O)2NH-alkyl, -S(O)2N(alkyl)2, cycloalkyl, cycloalkyloxy, heterocyclic, heterocyclic alkyloxy, heterocyclic alkyl, heterocyclic alkyloxy, heteroaryl, heteroaryloxy, aryl or aryloxy.

[0213] Unless otherwise specified, the term "3-6 membered heterocyclic alkyl" on its own or in combination with other terms refers to a saturated cyclic group consisting of 3 to 6 ring atoms, 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 carbon, nitrogen, and sulfur heteroatoms may optionally be oxidized (i.e., C(O), NO, and S(O)). p(where p is 1 or 2). It includes monocyclic and bicyclic systems, with bicyclic systems including spirocyclic, fused, and bridged rings. Furthermore, regarding the "3-6 membered heterocyclic alkyl," the heteroatom can occupy the connection position between the heterocyclic alkyl and the rest of the molecule. The 3-6 membered heterocyclic alkyl includes 4-6, 5-6, 4, 5, and 6 membered heterocyclic alkyls, etc. Examples of 3-6 membered heterocyclic alkyl groups include, but are not limited to, azirrobutyl, oxacyclobutyl, thiocyclobutyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiopheneyl (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxyl, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl or hexahydropyridazinyl, etc.

[0214] Unless otherwise specified, the term "5-6 membered heterocyclic alkyl" on its own or in combination with other terms refers to a saturated cyclic group consisting of 5 to 6 ring atoms, 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 carbon, nitrogen, and sulfur atoms may optionally be oxidized (i.e., C(O), NO, and S(O)). p (where p is 1 or 2). It includes monocyclic and bicyclic systems, wherein bicyclic systems include spirocyclic, fused, and bridged rings. Furthermore, regarding the "5-6 membered heterocyclic alkyl", the heteroatom can occupy the connection position between the heterocyclic alkyl group and the rest of the molecule. The 5-6 membered heterocyclic alkyl group includes 5-membered and 6-membered heterocyclic alkyl groups. Examples of 5-6 membered heterocyclic alkyl groups include, but are not limited to, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxyl, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl, hexahydropyridazinyl, etc.

[0215] Unless otherwise specified, the term "5-6 membered heterocyclic alkenyl" on its own or in combination with other terms respectively refers to a partially unsaturated cyclic group consisting of 5 to 6 ring atoms comprising at least one carbon-carbon double bond, 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 carbon, nitrogen, and sulfur atoms may optionally be oxidized (i.e., C(O), NO, and S(O)). p(where p is 1 or 2). It includes monocyclic and bicyclic systems, wherein bicyclic systems include spirocyclic, fused, and bridged rings, and any ring in this system is non-aromatic. Furthermore, regarding the "5-6 membered heterocyclic alkenyl," the heteroatom can occupy the connection position between the heterocyclic alkenyl and the rest of the molecule. The 5-6 membered heterocyclic alkenyl includes 5-membered and 6-membered heterocyclic alkenyl groups, etc. Examples of 5-6 membered heterocyclic alkenyl groups include, but are not limited to, those mentioned above.

[0216] The term "heteroaryl" refers to a monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, and S, with the remaining ring atoms being C, and having at least one aromatic ring. Preferred heteroaryls have a single 4- to 8-membered ring, particularly a 5- to 8-membered ring, or multiple fused rings containing 6 to 14, particularly 6 to 10 ring atoms. Non-limiting examples of heteroaryls include, but are not limited to, pyrroleyl, furanyl, thiopheneyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothiopheneyl, indoleyl, isoindoleyl, etc. The heteroaryl can be a 5- to 6-membered heteroaryl. The heteroaryl group is optionally substituted with one or more of the following groups: hydroxyl, amino, nitro, halogen, cyano, alkyl, alkenyl, alkoxy, haloalkoxy, alkylamino, dialkylamino, haloalkylamino, halodialkylamino, carboxyl, -C(O)O-alkyl, -OC(O)-alkyl, -C(O)NH2, -C(O)NH-alkyl, -C(O)N(alkyl)2, -NHC(O)-alkyl, -C(O)-alkyl, -S(O)-alkyl, -S(O)2-alkyl, -S(O)2NH2, -S(O)2NH-alkyl, -S(O)2N(alkyl)2, cycloalkyl, cycloalkyloxy, heterocyclic, heterocyclic alkyloxy, heterocyclic alkyl, heterocyclic alkyloxy, heteroaryl, heteroaryloxy, aryl or aryloxy.

[0217] Unless otherwise specified, the terms "5-6 membered heteroaryl" and "5-6 membered heteroaryl" are used interchangeably in this invention. The term "5-6 membered heteroaryl" refers to a monocyclic group with a conjugated π-electron system consisting of 5 to 6 ring atoms, 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. The nitrogen atom is optionally quaternized, and the carbon, nitrogen, and sulfur atoms may optionally be oxidized (i.e., C(O), 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.).

[0218] Unless otherwise specified, the terms "5-membered heteroaryl" and "5-membered heteroaryl" are used interchangeably in this invention. The term "5-membered heteroaryl" refers to a monocyclic group consisting of five ring atoms with a conjugated π-electron system, wherein one, two, three, or four ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. The nitrogen atom is optionally quaternized, and the carbon, nitrogen, and sulfur atoms may be optionally oxidized (i.e., C(O), NO, and S(O)). p (where p is 1 or 2). A 5-membered heteroaryl group can be attached to the rest of the molecule via a heteroatom or a carbon atom. Examples of such 5-membered heteroaryl groups 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-2H-3H-4H-3H-4H-5 ... H-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.).

[0219] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-12Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 C 11 and C 12 It also includes any range from n to n+m, such as C 1-12 Including C 1-3 C 1-6 C 1-9 C 3-6 C 3-9 C 3-12 C 6-9 C 6-12 and C 9-12 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-12-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, 11-membered, and 12-membered rings, and also any range from n to n+m. For example, 3-12-membered rings include 3-6-membered, 3-9-membered, 5-6-membered, 5-7-membered, 6-7-membered, 6-8-membered, and 6-10-membered rings, etc.

[0220] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (such as a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonates; chlorine, bromine, and iodine; sulfonate groups, such as methanesulfonates, toluenesulfonates, p-bromobenzenesulfonates, p-toluenesulfonates, etc.; acyloxy groups, such as acetoxy groups, trifluoroacetoxy groups, etc.

[0221] The term "protecting group" includes, but is not limited to, "amino protecting group," "hydroxy protecting group," or "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the nitrogen position of an amino group. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenemethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), triphenylmethyl (Tr), 1,1-di-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS), 2-(trimethylsilyl)ethoxymethyl (SEM), and tert-butyldimethylsilyl (TBS), etc. The term "hydroxyl protecting group" refers to a protecting group suitable for preventing hydroxyl side reactions. Representative hydroxyl protecting groups include, but are not limited to: alkyl groups, such as methyl, ethyl, and tert-butyl; acyl groups, such as alkanolyl groups (e.g., acetyl); arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (diphenylmethyl, DPM); silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc.

[0222] The compounds of the present invention can be prepared by a variety of synthetic methods 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 known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0223] The structures of the compounds of this invention can be confirmed by conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, that absolute configuration can be confirmed by conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) is used, where the cultured single crystal is used to collect diffraction intensity data using a Bruker D8 venture diffractometer with CuKα radiation as the light source. The scanning method is as follows: After scanning and collecting relevant data, the crystal structure can be further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0224] The volume used in this invention is commercially available.

[0225] The following abbreviations are used in this invention: DMAP represents 4-dimethylaminopyridine; DMSO represents dimethyl sulfoxide; CFU represents colony forming unit; DCM represents dichloromethane; DMF represents N,N-dimethylformamide; DME represents ethylene glycol dimethyl ether; TFA represents trifluoroacetic acid; Boc2O represents ditert-butyl dicarbonate; NaOH represents sodium hydroxide; NaH represents sodium hydride; NBS represents N-bromosuccinimide; TBSCl represents tert-butyldimethylchlorosilane; HPLC represents high-performance liquid chromatography; LCMS represents liquid chromatography-mass spectrometry; SBE-β-CD represents sulfobutyl ether β-cyclodextrin; PBS represents phosphate buffer.

[0226] Compounds are named according to conventional naming principles in the field or using Software naming conventions are used; commercially available compounds use supplier catalog names. Detailed Implementation

[0227] The present invention will be described in detail below with reference to examples, but this does not imply any adverse limitation on the invention. 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 embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the invention.

[0228] Example 1

[0229]

[0230] Synthesis route:

[0231]

[0232] Step 1: Preparation of Compounds 1-2

[0233] Compound 1-1 (5.00 g, 23.14 mmol) was dissolved in thionyl chloride (65.13 g, 547.43 mmol, 39.71 mL). The mixture was stirred under reflux for 2 hours. Thionyl chloride was removed by concentration under reduced pressure, and then methanol (50 mL) was added. The mixture was stirred at 65 °C for 1 hour. Methanol was removed by concentration under reduced pressure, and then the reaction mixture was quenched by adding an aqueous sodium carbonate solution (50 mL). Extraction was performed with ethyl acetate (50 mL × 2), the separated organic layer was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 1-2. LCMS (ESI) m / z: 229.9 / 231.9 (M+1).

[0234] Step 2: Preparation of compounds 1-3

[0235] Compounds 1-2 (15 g, 65.20 mmol), cyclopropylboronic acid (11.20 g, 130.4 mmol), copper acetate (23.68 g, 130.40 mmol), 2,2'-bipyridine (20.37 g, 130.40 mmol), and sodium carbonate (20.73 g, 195.60 mmol) were mixed in dichloromethane (240 mL), purged three times with O2, and then stirred at 15-25 °C under an oxygen atmosphere for 12 h. The reaction mixture was filtered, and the filtrate was added to water (200 mL) and extracted with dichloromethane (100 mL × 2). The separated organic layer was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to give compounds 1-3. LCMS(ESI)m / z:270.0 / 272.0(M+1).

[0236] Step 3: Preparation of compounds 1-4

[0237] Compounds 1-3 (1.8 g, 6.66 mmol), diphenylphosphine ferrocene palladium dichloride (487.59 mg, 666.37 μmol), bis-pinacol borate (2.03 g, 8.00 mmol), and potassium acetate (1.96 g, 19.99 mmol) were mixed in dioxane (30 mL), substituted with N2 three times, and then the mixture was stirred at 100 °C under a nitrogen atmosphere for 4 h. The reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 3), the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compounds 1-4. LCMS (ESI) m / z: 318.2 (M+1).

[0238] Step 4: Preparation of compounds 1-6

[0239] Compounds 1-5 (6.00 g, 25.97 mmol) were dissolved in dichloromethane (120 mL), and (Boc)₂O (14.17 g, 64.92 mmol) and DMAP (317.26 mg, 2.60 mmol) were added. The mixture was then stirred at 15–25 °C for 2 hours. The reaction solution was washed with saturated sodium bicarbonate aqueous solution (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compounds 1-6. LCMS (ESI) m / z: 431.0 / 433.0 (M+1).

[0240] Step 5: Preparation of compounds 1-7

[0241] Potassium carbonate (9.61 g, 69.56 mmol) was added to a methanol (45 mL) solution of compounds 1-6 (10.00 g, 23.19 mmol). The mixture was stirred at 15–25 °C for 2 hours. The methanol was removed by concentration under reduced pressure, and the reaction mixture was then quenched with water (50 mL). The mixture was extracted with ethyl acetate (50 mL × 2), the separated organic layer was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 0 to 2 / 1) to give compounds 1-7. 1 H NMR (400MHz, CDCl3) δ = 8.02-7.95 (m, 1H) 7.93-7.86 (m, 1H) 7.41 (br s, 1H), 3.97 (s, 3H), 1.51 (s, 9H).

[0242] Step Six: Preparation of Compounds 1-8

[0243] Compounds 1-7 (5.70 g, 17.21 mmol) were dissolved in acetone (85 mL), and methyl iodide (2.93 g, 20.65 mmol) and cesium carbonate (16.85 g, 51.64 mmol) were added. The mixture was then stirred at 15–25 °C for 3 hours. The reaction mixture was quenched with water (50 mL), extracted with ethyl acetate (50 mL × 2), and the separated organic layer was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compounds 1-8. LCMS (ESI) m / z: 344.9 / 446.9 (M+1).

[0244] Step 7: Preparation of compounds 1-9

[0245] Compounds 1-8 (1.8 g, 5.21 mmol), compounds 1-4 (2.1 g, 6.62 mmol), sodium carbonate (1.11 g, 10.43 mmol), diphenylphosphine ferrocene palladium dichloride (381.55 mg, 521.45 μmol), 2-dicyclohexylphosphine-2,6-dimethoxybiphenyl (214.07 mg, 521.45 μmol), and palladium acetate (58.54 mg, 260.73 μmol) were mixed in tetrahydrofuran (30 mL) and water (6 mL), and the mixture was substituted with N2 three times. The mixture was then stirred at 60-65 °C under a nitrogen atmosphere for 10 h. The reaction mixture was quenched with water (50 mL), extracted with ethyl acetate (30 mL × 3), and the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The residues were purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 5 / 1) to give compounds 1-9. LCMS (ESI) m / z: 456.1 (M+1).

[0246] Step 8: Preparation of compounds 1-10

[0247] Compounds 1-9 (0.50 g, 1.10 mmol) were dissolved in tetrahydrofuran (20 mL) at 0 °C, and lithium aluminum hydride (166.63 mg, 4.39 mmol) was added. The mixture was then stirred at 0 °C for 2 hours. The reaction mixture was quenched with water (1 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compounds 1-10. LCMS (ESI) m / z: 300.2 (M+1).

[0248] Step Nine: Preparation of Compounds 1-11

[0249] Compound 1-10 (0.30 g, 1.00 mmol) was dissolved in water (10 mL), and concentrated sulfuric acid (18.4 g, 183.85 mmol) was added. The mixture was then stirred at 60 °C for 12 hours. The reaction mixture was quenched with saturated sodium carbonate aqueous solution (100 mL), extracted with ethyl acetate (30 mL × 3), and the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 1-11. LCMS (ESI) m / z: 282.1 (M+1).

[0250] Step 10: Preparation of compounds 1-12

[0251] Compound 1-11 (0.26 g, 924.11 μmol) and diethyl 2-(ethoxymethylene)malonate (0.20 g, 924.11 μmol) were dissolved in toluene (5 mL) and purged three times with nitrogen. The mixture was stirred at 110 °C for 10 hours. The reaction mixture was concentrated under reduced pressure to give compound 1-12. LCMS (ESI) m / z: 452.1 (M+1).

[0252] Step 11: Preparation of compounds 1-13

[0253] Compounds 1-12 (0.12 g, 265.77 μmol) were dissolved in polyphosphoric acid (1.00 g). The mixture was then stirred at 90 °C for 2 hours. The reaction mixture was quenched with water (100 mL), the pH was adjusted to 8-9 with potassium carbonate, and then extracted with dichloromethane (20 mL × 4). The separated organic layer was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compounds 1-13. LCMS (ESI) m / z: 406.1 (M+1). Step Twelve: Preparation of Compound 1 Hydrochloride

[0254] Compounds 1-13 (0.05 g, 123.32 μmol) were dissolved in tetrahydrofuran (1 mL), ethanol (1 mL), and water (0.5 mL), and sodium hydroxide (14.80 mg, 369.96 μmol) was added. The mixture was then stirred at 15-25 °C for 2 hours. The pH of the reaction mixture was adjusted to 5-6 with 1 M hydrochloric acid, filtered, and concentrated under reduced pressure to obtain the residue. The hydrochloride salt of compound 1 was purified by preparative HPLC (column: YMCTriart 30 × 150 mm × 7 μm; mobile phase: 0.05% hydrochloric acid aqueous solution and acetonitrile; gradient: acetonitrile 12%-32%). 1H NMR (400MHz, CD3OD) δ = 9.08 (s, 1H), 8.64 (d, J = 8.4Hz, 1H), 8.39-8.28 (m, 1H), 7.89 (d, J = 8.5Hz, 1H), 7.26 ( d,J=9.4Hz,1H),4.89-4.88(m,4H),4.51-4.39(m,1H),3.18(s,3H),1.50-1.44(m,2H),1.09-1.03(m,2H). LCMS(ESI)m / z:378.1(M+1).

[0255] Example 2

[0256]

[0257] Synthesis route:

[0258]

[0259] Step 1: Preparation of Compound 2-2

[0260] Compound 2-1 (1.00 g, 4.33 mmol) was dissolved in dichloromethane (20 mL), and Boc₂O (2.36 g, 10.82 mmol) and DMAP (52.88 mg, 432.81 μmol) were added. The mixture was stirred at 20–25 °C for 2 hours. The reaction solution was washed successively with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 2-2. LCMS (ESI) m / z: 374.9 (M-56+1).

[0261] Step 2: Preparation of compounds 2-3

[0262] Potassium carbonate (1.44 g, 10.43 mmol) was added to a methanol (20 mL) solution of compound 2-2 (1.50 g, 3.48 mmol). The mixture was stirred at 20–25 °C for 2 hours. The crude product was filtered through a mixed solvent (petroleum ether / ethyl acetate = 1 / 1) under stirring to give compound 2-3. 1 H NMR (400MHz, CDCl3) δ = 8.47 (s, 1H), 8.33 (s, 1H), 7.68 (br s, 1H), 3.98 (s, 3H), 1.56 (s, 9H). LCMS(ESI)m / z:275.0(M-56+1).

[0263] Step 3: Preparation of compounds 2-4

[0264] Cesium carbonate (15.64 g, 48.01 mmol) and methyl iodide (2.73 g, 19.21 mmol) were added to an acetone (80 mL) solution of compounds 2-3 (5.3 g, 16.00 mmol). The mixture was stirred at 20–25 °C for 3 hours. The mixture was filtered and concentrated under reduced pressure to give the crude product. Compounds 2-4 were given by silica gel column chromatography (petroleum ether / ethyl acetate = 300 / 1 to 100 / 1). LCMS (ESI) m / z: 289.0 (M-56+1).

[0265] Step 4: Preparation of compounds 2-5

[0266] Compounds 1-4 (91.89 mg, 289.70 μmol), 2-dicyclohexylphosphonium-2,6-dimethoxybiphenyl (23.79 mg, 57.94 μmol), sodium carbonate (61.41 mg, 579.39 μmol), and palladium acetate (6.50 mg, 28.97 μmol) were added to a solution of compound 2-4 (0.1 g, 289.70 μmol) in tetrahydrofuran (2 mL) and water (0.4 mL). The mixture was purged with nitrogen three times. The mixture was heated to 70 °C and stirred for 22 hours. The mixture was filtered and concentrated under reduced pressure to give the crude compound. Compound 2-5 was obtained by thin-layer chromatography (petroleum ether / ethyl acetate = 5 / 1). LCMS (ESI) m / z: 456.1 (M+1).

[0267] Step 5: Preparation of compounds 2-6

[0268] At 0 °C, lithium aluminum hydride (316.63 mg, 8.34 mmol) was added to a tetrahydrofuran (20 mL) solution of compound 2-5 (950 mg, 2.09 mmol), and the mixture was stirred at 0–10 °C for 2 hours. The reaction solution was quenched with water (0.33 mL), 15% sodium hydroxide (0.33 mL), and water (1 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 2-6. LCMS (ESI) m / z: 344.2 (M-56+1).

[0269] Step Six: Preparation of Compounds 2-7

[0270] Compound 2-6 (500 mg, 1.67 mmol) was dissolved in 10 mL of 50% sulfuric acid aqueous solution, heated to 60 °C, and stirred for 12 hours. The reaction solution was diluted with 10 mL of water, the pH was adjusted to 9 with sodium carbonate, and extracted with 20 mL of ethyl acetate. Compound 2-7 was purified by thin-layer chromatography (petroleum ether / ethyl acetate = 1 / 1). LCMS (ESI) m / z: 282.2 (M+1).

[0271] Step 7: Preparation of compounds 2-8

[0272] Compound 2-7 (90.00 mg, 319.88 μmol) and diethyl 2-(ethoxymethylene)malonate (83.00 mg, 383.86 μmol) were dissolved in toluene (5 mL) at 25 °C, and the solution was heated to 110 °C and stirred for 14 hours. The solution was then concentrated to give compound 2-8. LCMS (ESI) m / z: 452.3 (M+1).

[0273] Step 8: Preparation of trifluoroacetate of compounds 2-9

[0274] Compounds 2-8 (0.15 g, 332.22 μmol) were dissolved in a polyphosphoric acid solution (88.59 μmol), heated to 90 °C, and stirred for 2 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (column: Shim-pack C18150*25 mm*10 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 12%-42%) to obtain the trifluoroacetate salt of compounds 2-9. LCMS (ESI) m / z: 406.3 (M+1).

[0275] Step Nine: Preparation of Compound 2 Trifluoroacetate

[0276] Sodium hydroxide (2.96 mg, 73.99 μmol) was added to a methanol (0.3 mL) and water (0.1 mL) solution of trifluoroacetate of compounds 2-9 (6 mg, 14.80 μmol) at 25 °C. The mixture was stirred at 25 °C for 2 hours, filtered, and the filtrate was purified by preparative HPLC (column: 3-Phenomenex Luna C1875*30 mm*3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 12%-42%) to obtain trifluoroacetate of compound 2. 1 H NMR (400MHz, CD3OD+D2O) δ = 9.15 (s, 1H), 8.59 (d, J = 8.4Hz, 1H), 8.31 (s, 1H), 7.89 (d, J = 8.4Hz, 1H), 7.2 3(s,1H),3.37(s,1H),3.32(s,4H),3.14(s,3H),2.72(s,1H),2.09(s,2H),1.58-1.44(m,2H),1.06(br d,J=2.3Hz,2H). LCMS(ESI)m / z:378.2(M+1).

[0277] Example 3

[0278] Synthesis route:

[0279]

[0280] Step 1: Preparation of compound 3-2

[0281] Compound 3-1 (5.00 g, 23.69 mmol) was dissolved in tert-butanol (60 mL) and water (60 mL), and potassium permanganate (18.72 g, 118.45 mmol) was added. The mixture was then stirred at 80 °C for 6 hours. The reaction solution was cooled to room temperature, filtered, and washed with water (20 mL) and methyl tert-butyl ether (50 mL), respectively. The aqueous phase was separated and the pH was adjusted to 4 with 1 M hydrochloric acid. The solution was concentrated under reduced pressure to give compound 3-2. LCMS (ESI) m / z: 241.0 / 243.0 (M+1).

[0282] Step 2: Preparation of compound 3-3

[0283] Thionyl chloride (4.94 g, 41.49 mmol) was added dropwise to a methanol (100 mL) solution of compound 3-2 (5 g, 20.74 mmol). The mixture was stirred at 70 °C for 12 hours. The reaction mixture was then quenched with water (30 mL), and the methanol was removed by concentration under reduced pressure. The mixture was extracted with ethyl acetate (50 mL), and the separated organic layers were washed with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 3-3. LCMS (ESI) m / z: 255.1 / 257.1 (M+1).

[0284] Step 3: Preparation of compounds 3-4

[0285] Compound 3-3 (1.00 g, 3.92 mmol) was dissolved in DMF (20 mL). Sodium hydride (235.21 mg, 5.88 mmol, 60% purity) was added at 0 °C, and the mixture was stirred at 0 °C for 1 hour. Then, 2-(trimethylsilyl)ethoxymethyl chloride (980.45 mg, 5.88 mmol) was added, and the mixture was stirred at 20-30 °C for 2 hours. The reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (20 mL × 2), and the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to give compound 3-4. LCMS (ESI) m / z: 515.1 / 517.1 (M+1).

[0286] Step 4: Preparation of compounds 3-5

[0287] Compounds 3-4 (0.19 g, 493.09 μmol), compounds 1-4 (187.68 mg, 591.70 μmol), sodium carbonate (130.66 mg, 1.23 μmol), and diphenylphosphine ferrocene palladium dichloride (36.08 mg, 49.31 μmol) were mixed in dioxane (4 mL) and water (1 mL), and the mixture was stirred in a microwave oven at 115 °C for 1 h. The reaction mixture was quenched with water (30 mL), extracted with ethyl acetate (30 mL × 2), the separated organic layer was washed with 20 mL of brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to give compounds 3-5. LCMS (ESI) m / z: 496.3 (M+1).

[0288] Step 5: Preparation of compounds 3-6

[0289] Compound 3-5 (0.55 g, 1.11 mmol) was dissolved in tetrahydrofuran (50 mL) at 0 °C, and lithium aluminum hydride (168.45 mg, 4.44 mmol) was added. The mixture was then stirred at 0 °C for 2 hours. The reaction mixture was quenched with water (4 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 3-6. LCMS (ESI) m / z: 440.2 (M+1).

[0290] Step Six: Preparation of Compounds 3-7

[0291] Compounds 3-6 (0.45 g, 1.02 mmol) were dissolved in water (5 mL), and concentrated sulfuric acid (9.2 g, 91.93 mmol) was added. The mixture was then stirred at 60 °C for 12 hours. The reaction mixture was quenched with an aqueous sodium carbonate solution (100 mL), extracted with ethyl acetate (30 mL × 3), and the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compounds 3-7. LCMS (ESI) m / z: 292.0 (M+1).

[0292] Step 7: Preparation of compounds 3-8

[0293] Compounds 3-7 (0.2 g, 686.47 μmol) and diethyl 2-(ethoxymethylene)malonate (178.12 mg, 823.76 μmol) were dissolved in N-methylpyrrolidone (4 mL), degassed, and purged three times with nitrogen. The mixture was stirred at 110 °C for 16 hours. The reaction mixture was quenched with water (30 mL), extracted with ethyl acetate (30 mL × 2), and the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give compounds 3-8. LCMS (ESI) m / z: 462.1 (M+1).

[0294] Step 8: Preparation of compounds 3-9

[0295] Compounds 3-8 (0.12 g, 260.02 μmol) were dissolved in polyphosphoric acid (5.00 g). The mixture was then stirred at 80 °C for 4 hours. The reaction mixture was quenched with saturated sodium carbonate aqueous solution (100 mL), extracted with ethyl acetate (30 mL × 2), and the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compounds 3-9. LCMS (ESI) m / z: 416.3 (M+1).

[0296] Step Nine: Preparation of Compound 3

[0297] Compounds 3-9 (0.02 g, 48.14 μmol) were dissolved in methanol (3 mL) and water (1 mL), and lithium hydroxide monohydrate (4.04 mg, 96.28 μmol) was added. The mixture was then stirred at 20-30 °C for 1 hour. The pH of the reaction mixture was adjusted to 5-6 with 1 M hydrochloric acid, and then concentrated under reduced pressure to obtain the residue. Compound 3 was purified by preparative HPLC (column: Phenomenex Synergi C18 150 × 25 mm × 10 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile: 8%-38%) to obtain compound 3. 1 H NMR (400MHz, DMSO-d6) δ = 8.91 (s, 1H), 8.57-8.46 (m, 1H), 8.40 (s, 1H), 8.09-7.76 ( m,3H),4.67-4.46(m,4H),3.98-3.98(m,1H),1.39-1.29(m,2H),1.08-1.01(m,2H). LCMS(ESI)m / z:388.1(M+1).

[0298] Example 4

[0299] Synthesis route:

[0300]

[0301] Step 1: Preparation of compound 4-2

[0302] Compound 4-1 (10.00 g, 48.20 mmol) was dissolved in dichloromethane (200 mL), and (Boc)₂O (26.30 g, 120.51 mmol, 27.69 mL) and DMAP (588.90 mg, 4.82 mmol) were added. The mixture was then stirred at 15–25 °C for 2 hours. The reaction solution was washed with saturated sodium bicarbonate aqueous solution (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 4-2. LCMS (ESI) m / z: 250.9 / 252.9 (M-100-56+1).

[0303] Step 2: Preparation of compound 4-3

[0304] Potassium carbonate (20.34 g, 147.17 mmol) was added to a methanol (100 mL) solution of compound 4-2 (20.00 g, 49.06 mmol). The mixture was stirred at 20–30 °C for 2 hours. The methanol was removed by concentration under reduced pressure, and the reaction mixture was quenched by adding water (100 mL) and ethyl acetate (50 mL). The mixture was filtered, and filter cake 1 was collected. The filtrate was extracted with ethyl acetate (30 mL × 2), and the separated organic layer was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was added to ethyl acetate (15 mL), stirred, filtered, and filter cake 2 was collected. The two filter cakes were combined and dried to give compound 4-3. LCMS (ESI) m / z: 250.9 / 252.9 (M-56+1).

[0305] Step 3: Preparation of compound 4-4

[0306] Compound 4-3 (13.2 g, 42.92 mmol) was dissolved in DMF (100 mL), and methyl iodide (7.31 g, 51.50 mmol) and cesium carbonate (41.95 g, 128.75 mmol) were added. The mixture was then stirred at 20–30 °C for 3 hours. The reaction mixture was quenched with water (200 mL), extracted with ethyl acetate (100 mL × 2), and the separated organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 4-4. LCMS (ESI) m / z: 264.9 / 266.9 (M-56+1).

[0307] Step 4: Preparation of compounds 4-5

[0308] Compound 4-4 (4.94 g, 14.93 mmol), allyltributyltin (4 g, 12.44 mmol), cesium fluoride (4.53 g, 29.85 mmol), and tetraphenylphosphine palladium (1.15 g, 995.03 μmol) were mixed in dioxane (40 mL), substituted with N2 three times, and then the mixture was stirred at 70 °C under a nitrogen atmosphere for 5 h. The reaction mixture was quenched with potassium fluoride aqueous solution (50 mL), stirred for 1 h, filtered, and the filtrate was extracted with ethyl acetate (50 mL × 3). The separated organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 10 / 1) to give compound 4-5. LCMS (ESI) m / z: 227.1 / 229.1 (M-56+1).

[0309] Step 5: Preparation of compounds 4-7

[0310] Compounds 4-6 (1.00 g, 3.27 mmol) were dissolved in chloroform (10 mL), and N-bromosuccinimide (698.53 mg, 3.92 mmol) and azobisisobutyronitrile (53.71 mg, 0.33 mmol) were added. The mixture was stirred at 80 °C for 16 hours. The reaction solution was then concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1 to 1 / 2) to give compounds 4-7. 1 H NMR (400MHz, CD3OD) δppm=8.90 (s, 1H), 8.36 (d, J = 8.75Hz, 1H), 7.61 (d, J = 8.76Hz, 1H), 4.32-4.41 (m, 3H), 2.71 (s, 2H), 1.40 (t, J = 7.07Hz, 7H).

[0311] Step Six: Preparation of Compounds 4-8

[0312] Compound 4-7 (2.2 g, 5.72 mmol) was dissolved in acetonitrile (60 mL) and then added... Molecular sieve (4.4 g) and N-methylmorpholine oxide (1.34 g, 11.44 mmol). The mixture was stirred at 20–30 °C for 2 hours. The reaction mixture was quenched with water (100 mL), extracted with ethyl acetate (50 mL × 2), the separated organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compounds 4–8. LCMS (ESI) m / z: 320.1 (M+1).

[0313] Step 7: Preparation of compounds 4-9

[0314] Methyltriphenylphosphine bromide (2.85 g, 7.98 mmol) was dissolved in tetrahydrofuran (80 mL), and then potassium tert-butoxide solution (1 M, 7.98 mL) was added at 0 °C. The mixture was stirred at 15–25 °C for 0.5 h. Then, compounds 4–8 (1.7 g, 5.32 mmol) were added, and the mixture was stirred at 15–25 °C for 2 h. The reaction mixture was quenched with water (200 mL), extracted with ethyl acetate (50 mL × 3), and the separated organic layer was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to give compounds 4–9. LCMS (ESI) m / z: 318.1 (M+1).

[0315] Step 8: Preparation of Compounds 4-10

[0316] Compounds 4-9 (1.5 g, 4.72 mmol), 2-di-tert-butylphosphine-2',4',6'-triisopropylbiphenyl (225.03 mg, 472.05 μmol), tricyclohexylphosphine (132.38 mg, 472.05 μmol), bis(pinacolborate) (1.80 g, 7.08 mmol), potassium acetate (926.53 mg, 9.44 mmol), and tris(dibenzylacetone)palladium (432.26 mg, 472.05 μmol) were mixed in dioxane (10 mL), substituted with N2 three times, and then the mixture was stirred at 100 °C under a nitrogen atmosphere for 2 h. The reaction mixture was quenched with water (50 mL), extracted with ethyl acetate (30 mL × 3), the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 1 / 1) to give compound 4-10. LCMS (ESI) m / z: 410.3 (M+1).

[0317] Step Nine: Preparation of Compound 4-11

[0318] Compound 4-10 (0.5 g, 1.22 mmol), compound 4-5 (345.44 mg, 1.12 mmol), potassium carbonate (337.68 g, 2.44 mmol), 2-di-tert-butylphosphine-2',4',6'-triisopropylbiphenyl (58.24 mg, 122.17 μmol), tricyclohexylphosphine (34.26 mg, 122.17 μmol), and tris(dibenzylideneacetone)palladium (111.87 mg, 122.17 μmol) were mixed in dioxane (10 mL) and water (2 mL), and the mixture was replaced with N2 three times. The mixture was then stirred at 100 °C under a nitrogen atmosphere for 1.5 h. The reaction mixture was quenched with water (50 mL), extracted with ethyl acetate (30 mL × 3), and the separated organic layer was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. Compound 4-11 was purified by preparative HPLC (column: Phenomenex luna C18 150 × 40 mm × 15 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile: 57%-87%). LCMS (ESI) m / z: 530.2 (M+1). 1 H NMR (400MHz, CDCl3) δ = 8.70 (s, 1H), 8.47 (d, J = 8.3Hz, 1H), 8.13-8.06 (m, 1H), 7.66 (s, 1H), 7.2 2(d,J=8.0Hz,1H),6.92-6.81(m,1H),5.75(tdd,J=6.6,10.2,16.9Hz,1H),5.34-5.26(m,1H),5 0.09-4.83 (m, 1H), 4.43 (q, J = 7.0 Hz, 2H), 3.84-3.74 (m, 1H), 3.45 (s, 3H), 3.18-3.00 (m, 2H), 1.58-1.55 (m, 9H), 1.44 (t, J = 7.1 Hz, 3H), 1.30-1.24 (m, 3H), 1.15-1.08 (m, 2H), 0.91-0.84 (m, 2H). Step 10: Preparation of compound 4-12

[0319] Compound 4-11 (0.04 g, 75.53 μmol) was dissolved in dichloromethane (2 mL), and Hoveyda-Grubbs second-generation catalyst (4.73 mg, 7.55 μmol) was added. The mixture was then stirred at 40 °C for 12 hours. The reaction mixture was concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 to 0 / 1) to give compound 4-12. LCMS (ESI) m / z: 502.3 (M+1).

[0320] Step 11: Preparation of Compound 4-13

[0321] Compound 4-12 (0.05 g, 99.69 μmol) was dissolved in dioxane (0.5 mL), and a hydrochloric acid / dioxane solution (4 M, 0.5 mL) was added. The mixture was then stirred at 10–20 °C for 2 hours. The reaction mixture was concentrated under reduced pressure to give compound 4-13. LCMS (ESI) m / z: 402.2 (M+1).

[0322] Step 12: Preparation of Compound 4

[0323] Compound 4-13 (0.04 g, 99.64 μmol) was dissolved in methanol (1 mL) and water (0.2 mL), and sodium hydroxide (7.97 mg, 199.27 μmol) was added. The mixture was then stirred at 10-20 °C for 2 hours. The pH of the reaction mixture was adjusted to 5-6 with acetic acid, and then concentrated under reduced pressure to obtain the residue. Compound 4 was purified by preparative HPLC (column: Phenomenex luna C18 150 × 25 mm × 10 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile: 4-34%) to obtain compound 4. 1 HNMR (400MHz, DMSO-d6) δ=15.09(s,1H),8.89(s,1H),8.38(s,1H),8.28(d,J=8.8Hz,1H),7.93(d,J=8.8Hz,1H),7.08(d,J=9.8Hz,1H),6.86(br d,J=4.9Hz,1H),6.39(s,1H),6.16(td,J=7.1,9.8Hz,1H),4.37(br t,J=4.4Hz,1H),3.45-3.39(m,2H),2.83(d,J=4.9Hz,3H),1.28-0.77(m,4H). LCMS(ESI)m / z:374.1(M+1).

[0324] Example 5

[0325] Synthesis route:

[0326]

[0327] Step 1: Preparation of Compound 5-2

[0328] Silver carbonate (5.41 g, 19.61 mmol) was added to a toluene (50 mL) solution of compound 5-1 (3.50 g, 15.08 mmol) and iodomethane (3.21 g, 22.63 mmol, 1.41 mL). The mixture was stirred at 100 °C for 1.5 h. The mixture was then cooled to room temperature, diluted with ethyl acetate (50 mL), filtered, and the mother liquor was concentrated to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 10 / 1) to give compound 5-2. LCMS (ESI) m / z: 246 / 248 (M+1); 1 H NMR (400MHz, CDCl3) δ = 8.37 (s, 1H), 7.10 (s, 1H), 3.97 (s, 3H), 3.95 (s, 3H).

[0329] Step 2: Preparation of compound 5-3

[0330] At 0 °C, lithium aluminum hydride (466.61 mg, 12.29 mmol) was added fractionally to a solution of 5-2 (2.75 g, 11.18 mmol) in 40 mL of tetrahydrofuran. The mixture was stirred at 0 °C for 0.5 h. The reaction mixture was quenched at 0 °C by adding sodium sulfate decahydrate (5.00 g), and then filtered. The mother liquor obtained from the filtration was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1 to 3 / 1) to give compound 5-3. LCMS (ESI) m / z: 218 / 220 (M+1).

[0331] Step 3: Preparation of compound 5-4

[0332] Sodium hydride (238.46 mg, 5.96 mmol, 60% purity) was added to a solution of compound 5-3 (1.00 g, 4.59 mmol) in tetrahydrofuran (15 mL) at 0 °C, followed by the addition of compound 4-7 (1.76 g, 4.59 mmol). The mixture was stirred at 0 °C for 3.5 h. The reaction was quenched with 15 mL of saturated ammonium chloride aqueous solution, and the mixture was extracted with ethyl acetate (15 mL × 2). The combined organic layers were washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1 to 1 / 1) to give compound 5-4. LCMS (ESI) m / z: 521 / 523 (M+1).

[0333] Step 4: Preparation of compound 5-5

[0334] Compound 5-4 (0.1 g, 0.19 mmol), bis(pinacolborate) borate (97.33 mg, 0.38 mmol), potassium acetate (37.62 g, 0.38 mmol), and bis(triphenylphosphine) ferrocene palladium dichloride (14.02 mg, 19.16 μmol) were mixed in ethylene glycol dimethyl ether (2 mL), purged three times with N2, and then the mixture was stirred at 80 °C under a nitrogen atmosphere for 16 h. Bis(triphenylphosphine) ferrocene palladium dichloride (14.02 mg, 19.16 μmol), sodium carbonate (40.62 mg, 0.38 mmol), and water (0.2 mL) were then added to the reaction mixture, and the mixture was stirred at 80 °C under a nitrogen atmosphere for 20 h. The reaction mixture was concentrated under reduced pressure to remove the solvent, yielding a residue. Compound 5-5 was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1 to 0 / 1) to give compound 5-5. LCMS(ESI)m / z:407(M+1).

[0335] Step 5: Preparation of Compound 5

[0336] At 20 °C, lithium hydroxide monohydrate (10.32 mg, 246.04 μmol) was added to a methanol (1 mL) and water (0.25 mL) solution of compound 5-5 (25 mg, 61.51 μmol). The mixture was stirred at 20 °C for 3 h, concentrated under reduced pressure, and the residue was purified by preparative HPLC (column: Phenomenex C1875 × 30 mm × 3 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile: 32%-62%) to obtain compound 5. LCMS (ESI) m / z: 309 (M+1); 1 H NMR (400MHz, DMSO-d6)δ=8.88-8.97(m,1H),8.61-8.68(m,1H),8.44-8.53(m,1H),7.90-8.01(m,1H),7 .10-7.17(m,1H),4.66-4.78(m,2H),4.44-4.59(m,3H),3.95-4.02(m,3H),1.30-1.38(m,2H),1.04(br d,J=8.19Hz,2H).

[0337] Example 6

[0338] Synthesis route:

[0339]

[0340] Step 1: Preparation of Compound 6-2

[0341] Compound 6-1 (1.72 g, 13.86 mmol) was dissolved in acetonitrile (17.2 mL), and NBS (2.47 g, 13.86 mmol) was added at 0–20 °C. The mixture was stirred for 3 hours under a nitrogen atmosphere. The reaction solution was quenched with water (20 mL) and diluted with ethyl acetate (20 mL). After dilution, the mixture was extracted with ethyl acetate (20 mL × 2). The mixed organic phase was washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to give compound 6-2. LCMS (ESI) m / z: 203.05 (M+1). 1 H NMR (400MHz, DMSO-d6) δ = 7.87 (s, 1H), 6.67 (s, 1H), 6.11 (s, 2H), 5.46 (t, J = 5.6Hz, 1H), 4.35 (d, J = 5.6Hz, 2H).

[0342] Step 2: Preparation of compound 6-3

[0343] Compound 6-2 (30 g, 147.76 mmol) and imidazole (15.09 g, 221.64 mmol) were dissolved in DMF (250 mL). Then, TBSCl (33.40 g, 221.64 mmol, 27.16 mL) was dissolved in DMF (50 mL). The mixture was slowly added dropwise to the reaction solution over 20 minutes at 25 °C, and stirred at 25 °C for 3 hours. The reaction solution was diluted with water (100 mL) and extracted with ethyl acetate (50 mL × 2). The mixed organic phase was washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was obtained by stirring in petroleum ether (100 mL), filtering, and drying the filter cake to give compound 6-3. LCMS (ESI) m / z: 319.1 (M+1).

[0344] Step 3: Preparation of compound 6-4

[0345] In a DMF (30 mL) solution of compound 6-3 (3 g, 9.45 mmol), NaH (1.51 g, 37.82 mmol) and methyl iodide (6.71 g, 47.27 mmol, 2.94 mL) were added sequentially. The mixture was stirred at 0 °C for 5 hours. The reaction solution was quenched with saturated ammonium chloride solution (50 mL), diluted with water (20 mL), extracted with ethyl acetate (20 mL × 2), washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Compound 6-4 was obtained by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1). LCMS (ESI) m / z: 346.9 (M+1).

[0346] Step 4: Preparation of Compound 6-5

[0347] A solution of compound 6-4 (1.92 g, 5.56 mmol) in dichloromethane (20 mL) was mixed with triethylamine hydrofluoric acid (4.48 g, 27.80 mmol, 4.53 mL) and stirred at 25°C under a nitrogen atmosphere for 2 hours. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL × 2). The mixed organic phase was washed with saturated brine (5 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude compound. The crude compound was then subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 5 / 1) to give compound 6-5. 1 H NMR (400MHz, DMSO-d6) δ = 8.09 (s, 1H), 6.83 (s, 1H), 4.44 (s, 2H), 3.05 (s, 6H).

[0348] Step 5: Preparation of compound 6-6

[0349] At 0 °C, sodium hydride (328.88 mg, 8.22 mmol, 60% purity) and compound 4-7 (1.58 g, 4.11 mmol) were added sequentially to a DMF (10 mL) solution of compound 6-5 (0.95 g, 4.11 mmol), and the mixture was stirred for 6 hours. The reaction solution was quenched with saturated ammonium chloride solution (10 mL), diluted with water (10 mL), extracted with ethyl acetate (10 mL × 2), and the mixed organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 1 / 2) to obtain compound 6-6. 1 H NMR (400MHz, DMSO-d6) δ = 8.60 (s, 1H), 8.14 (d, J = 8.6Hz, 1H), 8.02 (s, 1H), 7.57 (d, J = 8.7Hz, 1H), 6.51 (s,1H),5.39(s,2H),4.44(s,2H),4.24(q,J=7.1Hz,3H),2.94(s,6H),1.29(t,J=7.1Hz,3H),1.17(br d,J=7.1Hz,3H),0.90-0.85(m,2H).

[0350] Step Six: Preparation of Compounds 6-7

[0351] Compound 6-6 (0.5 g, 934.88 μmol), bis-pinacol borate (712.20 mg, 2.80 mmol), potassium pentovalinate (327.73 mg, 2.34 mmol), and tetra-triphenylphosphine palladium (108.03 mg, 93.49 μmol) were dissolved in ethylene glycol dimethyl ether (5 mL) at room temperature. The mixture was heated to 90 °C and stirred for 18 hours. The reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (20 mL). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 1 / 1) to give compound 6-7. LCMS (ESI) m / z: 582.3 (M+1).

[0352] Step 7: Preparation of trifluoroacetate of compound 6-8

[0353] Compounds 6-7 (390 mg, 670.23 μmol), potassium carbonate (231.58 mg, 1.68 mmol), tetraphenylphosphine palladium (24.71 mg, 67.02 μmol), and 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (63.90 mg, 134.05 μmol) were dissolved in water (0.4 mL) and ethylene glycol dimethyl ether (4 mL). The mixture was heated to 90 °C and stirred for 15 hours. The reaction solution was diluted with water (5 mL) and extracted with ethyl acetate (5 mL × 2). The mixed organic phase was washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25mm*4μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 24%-44%) to obtain trifluoroacetate salts of compounds 6-8. LCMS (ESI) m / z: 420.2 (M+1).

[0354] Step 8: Preparation of Compound 6 and its hydrochloride salt

[0355] Method 1: At 0℃, sodium hydroxide (14.30 mg, 357.59 μmol) was added to a mixture of methanol (0.6 mL) and water (0.3 mL) containing compounds 6-8 (30 mg, 71.52 μmol). The mixture was stirred at 20℃ under a nitrogen atmosphere for 15 hours. The solution was filtered, and the filtrate was purified by preparative HPLC (column: Waters Xbridge 150*25 mm*5 μm; mobile phase: 10 mM ammonium bicarbonate aqueous solution-acetonitrile; gradient: acetonitrile: 24%-54%) to obtain compound 6. LCMS (ESI) m / z: 392.1 (M+1). 1H NMR (400MHz, DMSO-d6) δ = 8.90 (s, 1H), 8.47 (d, J = 8.4Hz, 1H), 8.41 (s, 1H), 7.89 (d, J = 8.4Hz, 1H), 7.51 (s, 1H), 4.77 (br s,2H),4.56-4.52(m,2H),3.91-4.02(m,1H),3.15(s,6H),1.22-1.45(m,4H).

[0356] Method 2: At 0°C, 10 mL of an aqueous solution of sodium hydroxide (600.76 mg, 15.02 mmol) was added to 20 mL of methanol containing compound 6-8 (0.63 g, 1.50 mmol), and the mixture was stirred at 20-30°C for 15 hours. The mixture was filtered, and the filter cake was collected to obtain the crude product. At room temperature, the crude product was added to a mixture of hydrochloric acid (12 M, 122.63 μL) and an aqueous solution (4 mL), and the mixture was stirred for 16 hours. After filtration, the filter cake was collected and dried to obtain the hydrochloride salt of compound 6. LCMS (ESI) m / z: 392.1 (M+1).

[0357] Example 7

[0358]

[0359] Synthesis route:

[0360]

[0361] Step 1: Preparation of Compound 7-2

[0362] At 0 °C, sodium hydride (322.22 mg, 8.06 mmol) was added to a DMF (5 mL) solution of compound 7-1 (0.50 g, 4.03 mmol). After 10 minutes, compound 4-7 was added to the reaction solution and stirred for 6 hours. The reaction solution was quenched with saturated ammonium chloride solution (10 mL), extracted with ethyl acetate (10 mL), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Compound 7-2 was obtained by silica gel column chromatography (dichloromethane / methanol = 100 / 3). LCMS (ESI) m / z: 428.2 (M+1).

[0363] Step 2: Preparation of Compound 7-3

[0364] At 0 °C, bromosuccinimide (0.61 g, 3.41 mmol) was added to a solution of compound 7-2 (1.46 g, 3.41 mmol) in dichloromethane (20 mL), and the mixture was stirred at 0 °C for 2 hours. The reaction solution was concentrated, diluted with water (10 mL), and extracted with dichloromethane (10 mL × 2). The mixed organic phase was washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude compound. Compound 7-3 was obtained by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 0 to 100 / 3). LCMS (ESI) m / z: 508.1 (M+1).

[0365] Step 3: Preparation of compound 7-4

[0366] A solution of compound 7-3 (390.85 mg, 771.24 μmol) in dichloromethane (6 mL) was mixed with Boc₂O (0.42 mg, 1.93 mmol, 0.44 mL) and 4-dimethylaminopyridine (9.42 mg, 77.12 μmol) and stirred at 20–25 °C for 2 hours. The reaction mixture was diluted with water (5 mL) and extracted with dichloromethane (5 mL × 2). The mixed organic phase was washed with saturated brine (5 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 7-4. LCMS (ESI) m / z: 708.1 (M+1).

[0367] Step 4: Preparation of Compound 7-5

[0368] To a solution of compound 7-4 (0.05 g, 70.72 μmol) in ethylene glycol dimethyl ether (1 mL), bis-pinacol borate (35.92 mg, 141.44 μmol), potassium pentovalinate (24.79 mg, 176.80 μmol), and tetrakis(triphenylphosphine)palladium (2.61 mg, 7.07 μmol) were added sequentially, and the mixture was stirred at 90 °C for 15 hours. The reaction solution yielding crude compound 7-5 was used directly for the next step. LCMS (ESI) m / z: 672.3 (M+1).

[0369] Step 5: Preparation of Compounds 7-6

[0370] Crude compound 7-5 (47.52 mg, 70.72 μmol), potassium carbonate (24.44 mg, 176.80 μmol), tetraphenylphosphine palladium (2.61 mg, 7.07 μmol), and 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (6.74 mg, 14.14 μmol) were dissolved in water (0.1 mL) and ethylene glycol dimethyl ether (1 mL). The mixture was purged with nitrogen three times, heated to 90 °C, and stirred for 15 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain compound 7-6. LCMS (ESI) m / z: 592.2 (M+1). Step Six: Preparation of trifluoroacetate of compound 7-7 and compound 7-7.

[0371] A solution of compound 7-6 (41.84 mg, 70.72 μmol) in dichloromethane (1 mL) was mixed with trifluoroacetic acid (821.20 mg, 7.20 mmol, 533.25 μL) for 2 hours. Multiple batches were post-processed using two methods: Method 1: The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound 7-7; Method 2: The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then adjusted to pH 8.0 with saturated sodium carbonate solution, extracted with dichloromethane, and the organic phases were combined and concentrated under reduced pressure to obtain compound 7-7. LCMS (ESI) m / z: 392.1 (M+1).

[0372] Step 7: Preparation of trifluoroacetate and hydrochloride of compound 7

[0373] Method 1: Sodium hydroxide (26.52 mg, 662.97 μmol) was added to a solution of crude compound 7-7 (25.95 mg, 66.30 μmol) in methanol (1 mL) and water (0.5 mL) at 0 °C. The mixture was stirred at 20 °C for 15 hours under a nitrogen atmosphere. The crude compound was concentrated under reduced pressure and purified to obtain the trifluoroacetate of compound 7 by preparative HPLC (column: Phenomenex SynergiPolar-RP 100*25mm*4μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 17%-37%). LCMS (ESI) m / z: 364.1 (M+1). 1 H NMR (400MHz, DMSO-d6) δ = 8.90 (s, 1H), 8.47 (d, J = 8.4Hz, 1H), 8.41 (s, 1H), 7.89 (d, J = 8.4Hz, 1H), 6.91 (s, 1H), 4.77 (br s, 2H), 4.51 (s, 3H), 1.32 (br d,J=6.4Hz,2H),1.02(br s,2H).

[0374] Method 2: At room temperature, 5 mL of an aqueous solution of sodium hydroxide (102.18 mg, 2.55 mmol) was added to 10 mL of a methanol solution of crude compound 7-7 (1 g, 2.55 mmol), and the mixture was stirred at 20-50 °C for 36 hours. The mixture was filtered, and the filter cake was collected to obtain the crude product. 2 mL of hydrochloric acid (12 M) was added to 4 mL of the above aqueous solution of the crude product, and the mixture was stirred for 16 hours. After filtration, the filter cake was collected and dried to obtain the hydrochloride salt of compound 7. LCMS (ESI) m / z: 364.1 (M+1). 1 H NMR (400MHz, DMSO-d6) δ = 8.91 (s, 1H), 8.49 (d, J = 8.3Hz, 1H), 8.42 (s, 1H), 7.89 (d, J = 8.4Hz, 1H), 7.05(s,1H),4.81(s,2H),4.55(s,2H),4.52-4.43(m,1H),1.36-1.28(m,2H),1.07-0.99(m,2H).

[0375] Example 8

[0376]

[0377] Synthesis route:

[0378]

[0379] Step 1: Preparation of Compound 8-2

[0380] Compound 8-1 (1.00 g, 5.39 mmol) and N-methylbenzylamine (979.32 mg, 8.08 mmol, 1.04 mL) were dissolved in dioxane (20 mL) at 20 °C. Cesium carbonate (5.27 g, 16.16 mmol) and [tri-tert-butylphosphine-2-(2-aminobiphenyl)]palladium(II) chloride (276.07 mg, 538.77 μmol) were added. The mixture was stirred at 90 °C for 16 hours. The compound was filtered, concentrated, and subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1 to 30 / 1) to give compound 8-2. LCMS (ESI) m / z: 271.2 (M+1). 1 H NMR (400MHz, CDCl3) δ = 8.15 (d, J = 5.0Hz, 1H), 7.32-7.23 (m, 4H), 7.21-7.12 (m, 2H), 4.27 (s, 2H), 3.84 (s, 3H), 2.67 (s, 3H), 2.42 (s, 3H).

[0381] Step 2: Preparation of compound 8-3

[0382] Compound 8-2 (200 mg, 739.85 μmol) was dissolved in tetrahydrofuran (3 mL) solution, and lithium aluminum hydride (33.70 mg, 887.82 μmol) was added at 0 °C. The mixture was stirred at 0–25 °C for 2 hours. Water (0.034 mL) and 15% sodium hydroxide aqueous solution (0.034 mL) were added to the reaction solution, and the reaction was quenched by adding water (0.10 mL). Anhydrous magnesium sulfate was added, and the mixture was filtered and concentrated to obtain the crude compound. The crude compound was then subjected to silica gel column chromatography (petroleum ether / ethyl acetate: 3 / 1) to obtain compound 8-3. LCMS (ESI) m / z: 243.1 (M+1). 1 H NMR (400MHz, DMSO-d6) δ = 8.09 (d, J = 5.0Hz, 1H), 7.33 (d, J = 4.6Hz, 4H), 7.28-7.20 (m, 1H), 7.08 (d, J=5.0Hz,1H),5.29(t,J=5.4Hz,1H),4.50(d,J=5.4Hz,2H),4.22(s,2H),2.63(s,3H),2.19(s,3H).

[0383] Step 3: Preparation of compound 8-4

[0384] Compound 8-3 (800 mg, 3.30 mmol) was dissolved in a mixed solvent of ethanol (3 mL) and acetic acid (3 mL), and then 80 mg of Pd / C (10% purity) was added. A hydrogen balloon was placed over the reaction flask and purged three times to fill the reaction system with hydrogen gas. The mixture was stirred at 15-25 °C for 3 hours. The reaction solution was filtered, concentrated, and water (50 mL) was added. Extraction was performed with ethyl acetate (25 mL × 2). The organic phase was washed with water (15 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude compound 8-4. LCMS (ESI) m / z: 153.1 (M+1)

[0385] Step 4: Preparation of Compound 8-5

[0386] Compound 8-4 (156 mg, 1.03 mmol) was dissolved in DMF (1 mL). NaH (82.00 mg, 2.05 mmol, 60% purity) was added at 0–5 °C and the mixture was stirred for 0.5 hours. Then, compound 4-7 (413.99 mg, 1.08 mmol) was added, and the mixture was stirred at 0–5 °C for 3 hours. The reaction was quenched by adding saturated ammonium chloride aqueous solution (1 mL). Water (50 mL) was added, and the mixture was extracted with ethyl acetate (25 mL × 2). The organic phase was washed with saturated brine (25 mL × 2), filtered, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to give crude compound 8-5. LCMS (ESI) m / z: 456.1 (M+1).

[0387] Step 5: Preparation of compound 8-6 trifluoroacetate

[0388] Compound 8-5 (330 mg, 723.79 μmol) was dissolved in DCM (3 mL). N-bromosuccinimide (128.82 mg, 723.79 μmol) was added at 0 °C, and the mixture was stirred at 0–5 °C for 1 hour. The solution was concentrated, and the crude product was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 32%–52%) to obtain the trifluoroacetate salt of compound 8-6. LCMS (ESI) m / z: 536.0 (M+1).

[0389] Step Six: Preparation of Compounds 8-7

[0390] The trifluoroacetate of compounds 8-6 (90 mg, 168.28 μmol) and bis-pinacol boronic acid ester (85.46 mg, 336.56 μmol) were dissolved in ethylene glycol dimethyl ether (5 mL), followed by the addition of potassium pentovalinate (47.19 mg, 336.56 μmol) and tetraphenylphosphine palladium (19.45 mg, 16.83 μmol). Nitrogen gas was purged three times, and the mixture was stirred at 90 °C for 12 hours to obtain a crude reaction solution containing compounds 8-7, which was directly used in the next step. LCMS (ESI) m / z: 582.5 (M+1).

[0391] Step 7: Preparation of Compound 8-8

[0392] The crude reaction solution containing compound 8-7 (97.9 mg, 168.24 μmol) was added to potassium carbonate (46.51 mg, 336.49 μmol), tetratetraphenylphosphine palladium (19.44 mg, 16.82 μmol), and 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (16.04 mg, 33.65 μmol) dissolved in ethylene glycol dimethyl ether (2 mL) and water (0.2 mL). The reaction was carried out at 90 °C for 2 hours. The solution was concentrated, diluted with saturated brine (30 mL), extracted with ethyl acetate (30 mL × 2), washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude compound 8-8. LCMS (ESI) m / z: 420.2 (M+1).

[0393] Step 8: Preparation of compound 8 trifluoroacetate

[0394] Compound 8-8 (95 mg, 226.48 μmol) was dissolved in methanol (3 mL), then water (1 mL) and NaOH (54.35 mg, 1.36 mmol) were added, and the mixture was stirred at 15-20 °C for 12 hours. The mixture was filtered and concentrated to obtain a crude product. The crude product was purified by preparative HPLC (column: 3-Phenomenex Luna C1875*30 mm*3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 13%-33%) to obtain the trifluoroacetate salt of compound 8. LCMS (ESI) m / z: 392.1 (M+1). 1 HNMR (400MHz, CD3OD) δ = 9.09 (s, 1H), 8.64 (d, J = 8.3Hz, 1H), 8.15 (s, 1H), 7.85 (d, J = 8.3Hz, 1H), 4.99-4.95 (s, 2H) ,4.80-4.75(s,2H),4.45(tt,J=3.5,7.0Hz,1H),3.21(s,3H),2.42(s,3H),1.52-1.45(m,2H),1.12-1.03(m,2H).

[0395] Example 9

[0396]

[0397] Synthesis route:

[0398]

[0399] Step 1: Preparation of compound 9-2

[0400] Compound 9-1 (2 g, 13.06 mmol) was added to a mixed solvent of ethanol (125 mL) and dichloromethane (20 mL), followed by the addition of NaBH4 (1.51 g, 39.97 mmol). The mixture was stirred at 15–20 °C for 12 hours. The reaction solution was filtered, and the filtrate was concentrated. Acetone (20 mL) was added in an ice bath, followed by the addition of 2 M dilute hydrochloric acid until no more bubbles were produced. The pH was adjusted to 8 with saturated sodium bicarbonate solution, filtered, and the filtrate was concentrated. Ethanol (50 mL) was added, and the mixture was stirred at 15–20 °C for 0.5 hours. The mixture was then filtered, and the filtrate was concentrated to obtain compound 9-2. 1 H NMR (400MHz, DMSO-d6) δ = 8.20 (d, J = 4.9Hz, 1H), 6.66 (d, J = 5.0Hz, 1H), 6.50 (brs, 2H), 5.75-5.00 (br s, 1H), 4.30 (s, 2H).

[0401] Step 2: Preparation of compound 9-3

[0402] Compound 9-2 (0.87 g, 4.17 mmol) was dissolved in DMF (10 mL), and NaH (667.47 mg, 16.69 mmol) was added at 0 °C. After stirring for 0.5 hours, compound 4-7 (1.60 g, 4.17 mmol) was added, and the mixture was stirred at 0-5 °C for 3 hours. The mixture was then quenched with saturated brine (100 mL), dried over sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 9-3. LCMS (ESI) m / z: 429.1 (M+1).

[0403] Step 3: Preparation of compound 9-4

[0404] Compound 9-3 (0.55 g, 1.28 mmol) was dissolved in DCM (5 mL), and NBS (228.25 mg, 1.28 mmol) was added at 0 °C. The mixture was stirred for 1 hour. Compound 9-4 was obtained by silica gel rapid column chromatography (dichloromethane / methanol = 50 / 1). LCMS (ESI) m / z: 508.9 (M+1).

[0405] Step 4: Preparation of Compound 9-5

[0406] Compound 9-4 (0.8 g, 1.58 mmol), Boc₂O (1.20 g, 5.51 mmol, 1.27 mL), and DMAP (19.25 mg, 157.55 μmol) were dissolved in DCM (3 mL) and reacted with stirring at 10–25 °C for 12 hours. The reaction mixture was concentrated and compound 9-5 was obtained by silica gel rapid column chromatography (dichloromethane / methanol = 50 / 1). LCMS (ESI) m / z: 709.2 (M+1).

[0407] Step 5: Preparation of Compounds 9-6

[0408] Compound 9-5 (100 mg, 141.24 μmol), bis-pinacol borate (71.73 mg, 282.49 μmol), tetra-triphenylphosphine palladium (16.32 mg, 14.12 μmol), and potassium pentovalinate (39.61 mg, 282.49 μmol) were added to DME (2 mL). The mixture was stirred at 90 °C for 12 hours, cooled to room temperature, and then potassium carbonate (39.02 mg, 282.35 μmol), tetra-triphenylphosphine palladium (16.32 mg, 14.12 μmol), 2-dicyclohexylphosphine-2,4,6-triisopropylbiphenyl (13.46 mg, 28.24 μmol), and H₂O (0.2 mL) were added. The mixture was reacted at 90 °C for 2 hours. The reaction solution was concentrated to give compound 9-6. LCMS (ESI) m / z: 593.3 (M+1).

[0409] Step Six: Preparation of Compounds 9-7

[0410] Compound 9-6 (83 mg, 140.05 μmol) was added to a mixed solvent of MeOH (1 mL) and H₂O (3 mL), followed by sodium hydroxide (33.61 mg, 840.31 μmol). The mixture was stirred at 15-20 °C for 12 hours. The reaction solution was then concentrated to obtain compound 9-7. LCMS (ESI) m / z: 565.4 (M+1).

[0411] Step 7: Preparation of trifluoroacetate of compound 9

[0412] Compound 9-7 (79 mg, 139.93 μmol) was added to a mixed solvent of DCM (1 mL) and TFA (770.00 mg, 6.75 mmol) and stirred at 15-20 °C for 6 hours. The reaction solution was concentrated, and the crude product was dissolved in a mixed solvent of DMF (5 mL) and TFA (200 μL). The mixture was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: [0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 30%-50%). MeOH (1 mL) was added, and the mixture was stirred at 15-20 °C for 1 hour. The mixture was filtered, and the filter cake was washed with MeOH (0.5 mL) and dried under reduced pressure to obtain the trifluoroacetate salt of compound 9. LCMS (ESI) m / z: 365.1 (M+1).

[0413] Example 10

[0414]

[0415] Synthesis route:

[0416]

[0417] Step 1: Preparation of compound 10-1 trifluoroacetate

[0418] At 0 °C, compound 5-bromo-2-methylpyridine-4-methanol (525.28 mg, 2.60 mmol) was dissolved in DMF (10 mL), and NaH (415.96 mg, 10.40 mmol, 60% purity) was slowly added, and the reaction was allowed to proceed for 0.5 h. Then, compound 4-7 (1 g, 2.60 mmol) was added at 0 °C, and the mixture was stirred at 0 °C for 1 h. The reaction solution was quenched at 0 °C in a saturated ammonium chloride aqueous solution (20 mL), diluted with water (20 mL), extracted with ethyl acetate (90 mL), the organic layer was washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: Phenomenex Luna 150×40mm×15μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution and acetonitrile; the proportion of acetonitrile in the mobile phase ranged from 21% to 51%) to give the trifluoroacetate of compound 10-1. LCMS (ESI) m / z: 507.1 (M+1). 1HNMR(400MHz,CD3OD)δppm 0.81-0.92(m,2H)1.17(br d,J=6.24Hz,3H)1.26(t,J=7.09Hz,3H)2.42(s,3H)4.22(d,J=7.09Hz,3H)4.54(s,2H)5.4 9(s,2H)7.35(s,1H)7.42(d,J=8.68Hz,1H)8.13(d,J=8.68Hz,1H)8.47(s,1H)8.71(s,1H).

[0419] Step 2: Preparation of compound 10-2 trifluoroacetate

[0420] The trifluoroacetate of compound 10-1 (252 mg, 498.23 μmol), bis-pinacolborate (253.04 mg, 996.47 μmol), potassium pentovalinate (139.72 mg, 996.47 μmol), and triphenylphosphine palladium (36.74 mg, 99.65 μmol) were mixed in DMF (3 mL), degassed, and purged three times with nitrogen. The mixture was stirred at 120 °C under a nitrogen atmosphere for 16 h. The reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: 3-Phenomenex Luna 75 × 30 mm × 3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 15%-35%) to give compound 10-2 trifluoroacetate. LCMS (ESI) m / z: 391.2 (M+1). 1H NMR(400MHz,DMSO-d6)δppm 0.94(br s,2H)1.25-1.33(m,5H)2.65(s,3H)4.20-4.39(m,3H)4.57(s,2H)4.66(s,2H)7. 65(s,1H)7.83(d,J=8.31Hz,1H)8.38(d,J=8.19Hz,1H)8.67(s,1H)8.94(s,1H).

[0421] Step 3: Preparation of sodium salt of compound 10

[0422] At room temperature, a solution of sodium hydroxide (51.22 mg, 1.28 mmol) and water (0.3 mL) was added to a solution of 10⁻² trifluoroacetate (50 mg, 128.06 μmol) in anhydrous methanol (0.6 mL). The mixture was stirred at 20 °C for 3 h. The mixture was concentrated under reduced pressure to obtain a crude product. DMF (1 mL) was added to the crude product and stirred for 1 h. The mixture was filtered, and the filter cake was collected to obtain the sodium salt of compound 10. LCMS (ESI) m / z: 363.1 (M+1). 1 H NMR (400MHz, DMSO-d6) δppm 1.04 (br s, 2H) 1.34 (br d, J = 6.85Hz, 3H) 2.62 (s, 3H) 4.48-4.59 (m, 3H) 4.69 (br s,2H)7.54(s,1H)8.01(d,J=8.31Hz,1H)8.52(d,J=8.31Hz,1H)8.91(d,J=10.03Hz,2H)14.84(s,1H).

[0423] Example 11

[0424]

[0425] Synthesis route:

[0426]

[0427] Step 1: Preparation of trifluoroacetate of compound 11-1

[0428] At 0 °C, compound 3-bromopyridine-4-methanol (2.44 g, 12.98 mmol) was dissolved in DMF (30 mL), and NaH (2.08 g, 51.91 mmol, 60% purity) was slowly added, and the reaction was allowed to proceed for 0.5 h. Then, compound 4-7 (4.99 g, 12.98 mmol) was added at 0 °C, and the mixture was stirred at 0 °C for 1 h. The reaction mixture was quenched at 0 °C in saturated ammonium chloride aqueous solution (100 mL), diluted with water (50 mL), extracted with dichloromethane (150 mL), and the organic layer was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: Waters Xbridge 250 × 80 mm × 15 μm; mobile phase: 10 mmol / L trifluoroacetic acid aqueous solution and acetonitrile; gradient: acetonitrile 40%-70%) to obtain the trifluoroacetate of compound 11-1. 1 H NMR(400MHz,DMSO-d6)δppm 1.13-1.25(m,7H)4.39(s,2H)4.46(br s,1H)4.54(s,2H)5.46(s,2H)7.55(d,J=8.63Hz,1H)7.62(d,J=8.63Hz,1H)8.11(dd,J=8.57,5.19Hz,2H)8.60(s,1H)8.77(s,1H).

[0429] Step 2: Preparation of compound 11-2 trifluoroacetate

[0430] The trifluoroacetate of compound 11-1 (100 mg, 203.35 μmol), bis-pinacolborate (103.28 mg, 406.70 μmol), potassium pentovalinate (57.03 mg, 406.70 μmol), and triphenylphosphine palladium (15.00 mg, 40.67 μmol) were mixed in DMF (1 mL), degassed, and purged three times with nitrogen. The mixture was stirred at 120 °C under a nitrogen atmosphere for 16 h. After filtration, the filtrate was purified by preparative HPLC (column: 3-Phenomenex Luna 75 × 30 mm × 3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution and acetonitrile; gradient: acetonitrile 16%–36%) to give the trifluoroacetate of compound 11-2. LCMS (ESI) m / z: 377.1 (M+1).

[0431] Step 3: Preparation of Compound 11

[0432] At room temperature, sodium hydroxide (10.63 mg, 265.67 μmol) and water (0.3 mL) solution were added to an anhydrous methanol (0.6 mL) solution of trifluoroacetate 11-2 (10 mg, 26.57 μmol). The mixture was stirred at 20 °C for 3 h. After filtration, the filtrate was purified by preparative HPLC (column: 3-Phenomenex Luna 75 × 30 mm × 3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 15%-35%) to give trifluoroacetate of compound 11. LCMS (ESI) m / z: 349.0 (M+1). 1 H NMR (400MHz, CDCl3) δppm0.98-1.11(m,2H)1.22-1.27(m,2H)4.22(dt,J=7.21,3.48Hz,1H)4.59(s,2H)4.75(br s, 2H) 7.48 (d, J = 5.14Hz, 1H) 7.83 (d, J = 8.31Hz, 1H) 8.70 (d, J = 8.19Hz, 1H) 8.83 (d, J = 4.89Hz, 1H) 9.00 (d, J = 5.87Hz, 2H) 14.54 (br d, J = 3.55Hz, 1H).

[0433] Example 12

[0434]

[0435] Synthesis route:

[0436]

[0437] Step 1: Preparation of trifluoroacetate of compound 12-1 and compound 12-1

[0438] At room temperature, acetic acid (27.61 mg, 459.87 μmol) was added to a solution of trifluoroacetate of compound 7-7 (0.12 g, 306.58 μmol), tert-butyldimethylsiloxane acetaldehyde (106.88 mg, 613.15 μmol) in dichloromethane (3 mL) and ethanol (1.5 mL). The mixture was heated to 40 °C and stirred for 1 hour. Then, the mixture was cooled to room temperature and sodium cyanoborohydride (57.80 mg, 919.73 μmol) was added. The reaction was continued at 40 °C for 1 hour. Post-processing was performed on multiple batches. Method 1: The reaction solution was concentrated under reduced pressure and then purified by preparative HPLC (column: 3-Phenomenex Luna C1870*30mm*3μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution and acetonitrile; gradient: acetonitrile 37%-57%) to obtain compound 12-1 trifluoroacetate. Method 2: The reaction solution was concentrated under reduced pressure to obtain crude product, the pH was adjusted to 8.0 with saturated sodium carbonate solution, and after extraction with dichloromethane, the organic phases were combined and concentrated under reduced pressure to obtain compound 12-1. LCMS (ESI) m / z: 436.2 (M+1).

[0439] Step 2: Preparation of Compound 12 Trifluoroacetate and Compound 12 Hydrochloride

[0440] Method 1: At room temperature, 0.3 mL of an aqueous solution of sodium hydroxide (55.11 mg, 1.38 mmol) was added to 0.6 mL of methanol containing compound 12-1 (0.06 g, 137.78 μmol), and the mixture was stirred at room temperature for 3 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 20%-40%) to obtain the trifluoroacetate of compound 12. LCMS (ESI) m / z: 408.2 (M+1).

[0441] Method 2: At room temperature, 10 mL of an aqueous solution of sodium hydroxide (918.54 mg, 22.96 mmol) was added to 20 mL of a methanol solution of compound 12-1 (1 g, 2.30 mmol), and the mixture was stirred at 20-50 °C for 36 hours. The mixture was filtered, and the filter cake was collected to obtain the crude product. 2 mL of 12 M hydrochloric acid was added to 4 mL of an aqueous solution of the crude product (100 mg, 245.45 μmol), and the mixture was stirred for 16 hours. The reaction mixture was filtered, and the filter cake was collected and dried to obtain the hydrochloride salt of compound 12. LCMS (ESI) m / z: 408.1 (M+1). 1HNMR (400MHz, CD3OD) δppm 9.08 (s, 1H), 8.63 (d, J = 8.44Hz, 1H), 8.31 (s, 1H), 7.85 (d, J = 8.31Hz, 1H), 7.18 (s, 1H), 4.96 (br s,2H), 4.64(s,2H), 4.41-4.48(m,1H), 3.87(t,J=5.20Hz,2H), 3.63(t,J=5.32Hz,2H), 1.44-1.51(m,2H), 1.05-1.11(m,2H).

[0442] Example 13

[0443]

[0444] Synthesis route:

[0445]

[0446] Step 1: Preparation of trifluoroacetate of compound 13-1

[0447] At room temperature, acetic acid (4.60 mg, 76.64 μmol) was added to a solution of compound 7-7 (0.02 g, 51.10 μmol), propionaldehyde (4.45 mg, 76.64 μmol), and ethanol (0.5 mL). The mixture was heated to 40 °C and stirred for 1 hour. Then, sodium cyanoborohydride (9.63 mg, 153.29 μmol) was added to the solution, and the reaction was continued at 40 °C for 1 hour. The reaction solution was concentrated under reduced pressure and purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 28%-48%) to obtain the trifluoroacetate of compound 13-1. LCMS (ESI) m / z: 434.2 (M+1).

[0448] Step 2: Preparation of compound 13 trifluoroacetate

[0449] At room temperature, a solution of sodium hydroxide (5.07 mg, 126.87 μmol) in water (0.25 mL) was added to a solution of trifluoroacetate of compound 13-1 in methanol (0.5 mL), and the mixture was stirred at room temperature for 16 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 20%-40%) to obtain trifluoroacetate of compound 13. LCMS (ESI) m / z: 406.2 (M+1).

[0450] Example 14

[0451]

[0452] Synthesis route:

[0453]

[0454] Step 1: Preparation of Compound 14-1

[0455] At room temperature, acetic acid (3.38 mg, 76.65 μmol) was added to a solution of trifluoroacetate of compound 7-7 (0.02 g, 51.10 μmol), acetaldehyde (3.38 mg, 76.65 μmol), dichloromethane (1 mL), and ethanol (0.5 mL). The mixture was heated to 30 °C and stirred for 1 hour. Then, sodium cyanoborohydride (9.63 mg, 153.29 μmol) was added to the solution at room temperature, and the reaction was continued at 30 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 3-9. LCMS (ESI) m / z: 420.1 (M+1). Step 2: Preparation of trifluoroacetate of compound 14

[0456] At room temperature, 0.5 mL of an aqueous solution of sodium hydroxide (10.22 mg, 255.56 μmol) was added to 1 mL of methanol containing compound 14-1 (21.44 mg, 51.11 μmol), and the mixture was stirred at 30 °C for 16 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 23%-43%) to give the trifluoroacetate of compound 14. LCMS (ESI) m / z: 392.1 (M+1).

[0457] Example 15

[0458]

[0459] Synthesis route:

[0460]

[0461] Step 1: Preparation of Compound 15-1

[0462] At room temperature, acetic acid (6.90 mg, 114.97 μmol) was added to a solution of trifluoroacetate of compound 7-7 (0.03 g, 76.64 μmol), benzaldehyde (16.27 mg, 153.29 μmol) in dichloromethane (1 mL) and ethanol (0.5 mL). The mixture was heated to 30 °C and stirred for 1 hour. Then, sodium cyanoborohydride (9.63 mg, 153.29 μmol) was added at room temperature, and the reaction was carried out at 50 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain crude compound 15-1. LCMS (ESI) m / z: 482.3 (M+1). Step 2: Preparation of trifluoroacetate of compound 15

[0463] At room temperature, 1 mL of an aqueous solution of sodium hydroxide (24.92 mg, 623.00 μmol) was added to 2 mL of methanol containing compound 15-1 (60 mg, 124.60 μmol), and the mixture was stirred at 30 °C for 15 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 32%-52%) to obtain trifluoroacetic acid of compound 15. LCMS (ESI) m / z: 454.2 (M+1). 1HNMR (400MHz, CD3OD) δ = 9.08 (s, 1H), 8.64 (d, J = 8.3Hz, 1H), 8.31 (s, 1H), 7.85 (d, J = 8.3Hz, 1H), 7.54- 7.34(m,6H),7.26(s,1H),4.98(s,2H),4.72(s,2H),4.65(s,2H),4.44(td,J=3.5,7.2Hz,1H),1.47(br d,J=6.5Hz,2H),1.08(br d,J=2.6Hz,2H).

[0464] Example 16

[0465]

[0466] Synthesis route:

[0467]

[0468] Step 1: Preparation of Compound 16-1

[0469] At room temperature, acetic acid (23.01 mg, 383.22 μmol) was added to a solution of trifluoroacetate (0.1 g, 255.48 μmol) of cyclobutylcarboxaldehyde (42.98 mg, 510.96 μmol) in dichloromethane (10 mL) and methanol (5 mL). The mixture was heated to 40 °C and stirred for 1 hour. Then, the temperature was lowered to room temperature, and pyridineborane (71.23 mg, 766.44 μmol) was added. The reaction was continued at 40 °C for 40 hours. The reaction solution was quenched with saturated ammonium chloride solution (10 mL), extracted with dichloromethane (10 mL), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude product 16-1. LCMS (ESI) m / z: 460.2 (M+1).

[0470] Step 2: Preparation of compound 16 trifluoroacetate

[0471] At room temperature, 0.6 mL of an aqueous solution of sodium hydroxide (24.92 mg, 623.00 μmol) was added to 1.2 mL of methanol containing compound 16-1 (120 mg, 250.69 μmol), and the mixture was stirred at 40 °C for 16 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (column: 3-Phenomenex Luna C1875*30 mm*3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 29%-49%) to give the trifluoroacetate of compound 16. LCMS (ESI) m / z: 432.2 (M+1). 1H NMR (400MHz, DMSO-d6) δ=8.89(s,1H),8.45(d,J=8.3Hz,1H),8.37(s,1H),7.88(d,J=8.5Hz,1H),6.97(br s,1H),4.77(br s,2H),4.51(s,2H),4.48(br d,J=3.6Hz,1H),3.43(br d,J=7.1Hz,2H),2.68-2.59(m,1H),2.18-2.00(m,2H),1.95-1.84(m,2H),1.83-1.68(m,2H),1.32(br d,J=6.6Hz,2H),1.02(br s,2H).

[0472] Example 17

[0473]

[0474] Synthesis route:

[0475]

[0476] Step 1: Preparation of Compound 17-1

[0477] At room temperature, acetic acid (23.01 mg, 383.22 μmol) was added to a solution of trifluoroacetate (0.1 g, 255.48 μmol) of oxa-3-cyclobutylcarboxaldehyde (43.99 mg, 510.96 μmol) in dichloromethane (10 mL) and methanol (5 mL). The mixture was heated to 40 °C and stirred for 1 hour. Then, the temperature was lowered to room temperature, and pyridineborane (71.23 mg, 766.44 μmol) was added. The reaction was continued at 40 °C for 40 hours. The reaction solution was quenched with saturated ammonium chloride solution (10 mL), extracted with dichloromethane (10 mL), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 17-1. LCMS (ESI) m / z: 462.2 (M+1).

[0478] Step 2: Preparation of compound 17 trifluoroacetate

[0479] At room temperature, 0.5 mL of an aqueous solution of sodium hydroxide (8.67 mg, 216.68 μmol) was added to 1 mL of methanol containing 10 mg, 21.67 μmol of compound 17-1, and stirred at 40 °C for 16 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (column: 3-Phenomenex Luna C1875 * 30 mm * 3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 11%-31%) to give the trifluoroacetate of compound 17. LCMS (ESI) m / z: 434.1 (M+1). 1H NMR (400MHz, DMSO-d6) δ = 9.68 (br d, J = 1.1Hz, 1H), 8.93 (s, 1H), 8.54 (d, J = 2.2Hz, 1H), 7.92-7.83 (m, 1H), 7.16 (s, 1H), 4.91-4.81 (m, 2H), 4.58 (br d,J=1.3Hz,3H),4.53-4.43(m,2H),4.22-4.19(m,1H),3.58(br dd,J=3.2,6.9Hz,2H),3.54-3.46(m,1H),1.33-1.33(m,1H),1.31(br d,J=7.6Hz,1H),1.03(br s,2H).

[0480] Example 18

[0481]

[0482] Synthesis route:

[0483]

[0484] Step 1: Preparation of Compound 18-1

[0485] At 20 °C, trifluoroacetate of compound 7-7 (20 mg, 51.10 μmol) and N-Boc-3-formylazonium butane (14.20 mg, 76.64 μmol) were dissolved in dichloromethane (1 mL) and water (0.5 mL), followed by the addition of glacial acetic acid (4.60 mg, 76.64 μmol). The mixture was stirred at 50 °C for 16 hours. At room temperature, sodium cyanoborohydride (9.63 mg, 153.29 μmol) was added, and the mixture was stirred at 20 °C for 5 hours, then heated to 50 °C and stirred for 16 hours. The mixture was filtered and concentrated under reduced pressure to give compound 18-1. LCMS (ESI) m / z: 561.4 (M+1).

[0486] Step 2: Preparation of compound 18-2 trifluoroacetate

[0487] Compound 18-1 (80 mg, 142.69 μmol) was dissolved in methanol (2 mL) at 20 °C, and sodium hydroxide solution (28.54 mg sodium hydroxide dissolved in 1 mL water) was added. The mixture was stirred at 20 °C for 16 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 32%-52%) to give compound 18-2 trifluoroacetate. LCMS (ESI) m / z: 533.3 (M+1).

[0488] Step 3: Preparation of compound 18 trifluoroacetate

[0489] At 20 °C, the trifluoroacetate of compound 18-2 (42 mg, 78.86 μmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (616.00 mg, 5.40 mmol) was slowly added. The mixture was stirred at 20 °C for 2 hours. The mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (column: 3-Phenomenex Luna C18, 70*30 mm*3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 5%-25%) to obtain the trifluoroacetate of compound 18. LCMS (ESI) m / z: 433.2 (M+1). 1H NMR(400MHz,CD3OD)δppm 1.02-1.15(m,2H)1.23-1.40(m,2H)1.42-1.54(m,2H)3.17-3.28(m,2H)3.51(br dd,J=13.27,8.25Hz,2H)3.70-4.07(m,2H)4.43(td,J=7.27,3.91Hz,1H)4.62-4.69(m,2H)5.01(br s,2H)7.20(s,1H)7.84(br d,J=8.44Hz,1H)8.33(br s,1H)8.54-8.72(m,1H)9.07(br d,J=5.99Hz,1H).

[0490] Example 19

[0491]

[0492] Synthesis route:

[0493]

[0494] Step 1: Preparation of trifluoroacetate of compound 19-1

[0495] At room temperature, trifluoroacetate of compound 7-7 (55 mg, 140.51 μmol) and tert-butyl N-(2-oxycarbonylethyl)carbamate (22.37 mg, 140.51 μmol) were dissolved in dichloromethane (1 mL) and methanol (0.5 mL), and glacial acetic acid (16.88 mg, 281.03 μmol) was added. The mixture was stirred at 50 °C for 3 hours. Sodium cyanoborohydride (26.49 mg, 421.54 μmol) was added at 20 °C, and the mixture was stirred at 20 °C for 1 hour, then heated to 50 °C and stirred for 16 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: 3-Phenomenex Luna C18, 75 × 30 mm × 3 μm; mobile phase: 0.05% aqueous trifluoroacetic acid and acetonitrile; acetonitrile content in the mobile phase ranged from 25% to 45%) to give trifluoroacetate of compound 19-1. LCMS(ESI) m / z: 535.3(M+1).

[0496] Step 2: Preparation of compound 19-2 trifluoroacetic acid

[0497] At room temperature, 22 mg (41.15 μmol) of trifluoroacetate 19-1 was dissolved in 1 mL of dichloromethane, and trifluoroacetic acid (469.22 mg, 4.12 mmol) was added. The mixture was stirred at 20 °C for 2 hours. The mixture was concentrated under reduced pressure to give trifluoroacetate of compound 19-2. LCMS (ESI) m / z: 435.1 (M+1).

[0498] Step 3: Preparation of trifluoroacetate of compound 19

[0499] At room temperature, 17.8 mg (40.97 μmol) of trifluoroacetate 19-2 was dissolved in methanol (1 mL), and sodium hydroxide solution (8.19 mg dissolved in 0.5 mL water) was added. The mixture was stirred at 20 °C for 16 hours. The mixture was concentrated under reduced pressure to obtain a residue. The residue was purified by preparative HPLC (column: 3-Phenomenex Luna C18, 75 × 30 mm × 3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 10%-30%) to give trifluoroacetate of compound 19. LCMS (ESI) m / z: 407.1 (M+1). 1 H NMR(400MHz,CD3OD)δppm 0.90-1.01(m,2H)1.36(br d,J=6.97Hz,2H)3.13(t,J=5.69Hz,2H)3.64(t,J=5.69Hz,2H)4.27-4.36(m,1H)4.44(s,2H)4.7 2-4.75(m,2H)6.69(s,1H)7.73(d,J=8.44Hz,1H)8.37(s,1H)8.46(d,J=8.44Hz,1H)8.94(s,1H).

[0500] Example 20

[0501]

[0502] Synthesis route:

[0503]

[0504] Step 1: Preparation of Compound 20-1

[0505] Compound 4-11 (15 mg, 29.91 μmol) was dissolved in methanol (2 mL) at room temperature, and wet Pd / C (10 mg, 50% water content) was added. The mixture was stirred at 10-15 °C under a hydrogen (15 Psi) atmosphere for 2 hours, filtered, and concentrated under reduced pressure to obtain compound 20-1. LCMS (ESI) m / z: 504.2 (M+1).

[0506] Step 2: Preparation of compound 20-2 hydrochloride

[0507] At room temperature, compound 20-1 (16 mg, 28.59 μmol) was dissolved in methanol (1 mL) and water (0.5 mL), and sodium hydroxide (6.86 mg, 171.57 μmol) was added. The mixture was stirred at 10-15 °C for 5 hours. The pH of the reaction solution was adjusted to 1-2 with 1 M hydrochloric acid aqueous solution, and the solution was concentrated to obtain the hydrochloride salt of compound 20-2. LCMS (ESI) m / z: 476.1 (M+1). Step 3: Preparation of trifluoroacetate of compound 20

[0508] At room temperature, 14 mg (29.44 μmol) of the hydrochloride salt of compound 20-2 was dissolved in dichloromethane (1 mL), and 0.5 mL of trifluoroacetic acid was added. The mixture was stirred at 25-30 °C for 1 hour. The solution was concentrated, and the residue was purified by preparative HPLC (column: 3-Phenomenex Luna C1875*30 mm*3 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 18%-38%) to give the trifluoroacetate salt of compound 20. LCMS (ESI) m / z: 376.0 (M+1). 1 H NMR (400MHz, CDCl3) δ = 14.51 (s, 1H), 10.22 (s, 1H), 9.01 (s, 1H), 8.51 (s, 1H), 7.85 (s, 1H), 7.4 3(m,1H),6.68(s,1H),3.13(s,3H),2.74(s,2H),2.39(s,2H),1.36-1.31(m,2H),1.14(s,2H).

[0509] Example 21

[0510] Synthesis route:

[0511]

[0512] Step 1: Preparation of Compound 21

[0513] Compound 5-5 (105 mg, 258.35 μmol) was added to hydrobromic acid (12 mL, 40% aqueous solution) and stirred at 110 °C for 8 hours. The mixture was concentrated, and the residue was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile: 29%-49%) to give compound 21. LCMS (ESI) m / z: 365.3 (M+1). 1HNMR (400MHz, DMSO-d6) δ = 8.88 (s, 1H), 8.42-8.40 (m, 1H), 7.92 (s, 1H), 7.84-7.8 2(m,1H),6.60(s,1H),4.84(s,1H),4.44(s,1H),1.32-1.30(m,2H),1.00(s,2H).

[0514] Example 22

[0515]

[0516] Synthesis route:

[0517]

[0518] Step 1: Preparation of Compound 22-1

[0519] At room temperature, compound 7-7 (70.00 mg, 178.84 μmol) and (R)-(+)-2,2-dimethyl-1,3-dioxo-4-aldehyde pentane (46.55 mg, 357.67 μmol) were dissolved in anhydrous dichloromethane (4 mL) and anhydrous methanol (2 mL). Acetic acid (16.11 mg, 268.25 μmol) was added at 20-30 °C, and the mixture was heated to 40 °C and stirred for 1 hour. The temperature was then lowered to 20-30 °C, and pyridine borane (49.86 mg, 536.51 μmol) was added. The mixture was then heated to 40 °C and stirred for 16 hours. The reaction mixture was concentrated, and 10 mL of dichloromethane was added. The mixture was washed successively with saturated sodium bicarbonate (10 mL * 2) and brine (10 mL), dried, and concentrated to obtain compound 22-1. LCMS (ESI) m / z: 506.1 (M+1).

[0520] Step 2: Preparation of trifluoroacetate of compound 22

[0521] At room temperature, concentrated hydrochloric acid (0.5 mL) was added to a solution of compound 22-1 (35.00 mg, 69.23 μmol) in ethanol (1.5 mL) and water (1.0 mL). The reaction mixture was stirred at 40 °C for 22 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: YMC Triart C18150*25 mm*5 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution and acetonitrile; acetonitrile content in the mobile phase ranged from 11% to 31%, 10 min) to obtain the trifluoroacetate of compound 22.

[0522] Example 23

[0523]

[0524] Synthesis route:

[0525]

[0526] Step 1: Preparation of compounds 2-9

[0527] Compound 2-9 (50 mg, 123.32 μmol) was dissolved in anhydrous N,N-dimethylformamide (3 mL) at room temperature. After the reaction solution was cooled to 0 °C, sodium hydride (5.43 mg, 135.65 μmol) and difluoroethyl trifluoromethanesulfonate (29.04 mg, 135.65 μmol) were added sequentially under nitrogen protection. The reaction solution was stirred at 25 °C for 1 hour. No post-treatment was required to obtain compound 23-1. LCMS (ESI) m / z: 470.2 (M+1).

[0528] Step 2: Preparation of trifluoroacetate of compound 23

[0529] At room temperature, a solution of sodium hydroxide (4.86 mg, 121.41 μmol) in anhydrous N,N-dimethylformamide (3 mL) was added to a solution of sodium hydroxide (4.86 mg, 121.41 μmol) in water (1 mL). The reaction mixture was stirred at 25 °C for 3 hours. The crude product was obtained by concentration under reduced pressure. The crude product was then purified by preparative HPLC (column: YMC Triart C18150*25 mm*5 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution and acetonitrile; acetonitrile content in the mobile phase ranged from 39% to 59%, 10 min) to obtain the trifluoroacetate of compound 23. LCMS (ESI) m / z: 442.1 (M+1). 1 H NMR(400MHz,DMSO-d6)δppm 14.83-15.13(m,1H), 8.85-8.94(m,1H), 8.54-8.62(m,1H), 8.41-8.48(m,1H), 7.88-7.99(m,1H), 6.99-7.07(m,1H), 6.06-6.48(m,1H) , 4.65-4.74(m,2H), 4.52-4.56(m,2H), 4.45-4.50(m,1H), 4.05-4.18(m,2H), 3.13-3.24(m,3H), 1.28-1.38(m,2H), 0.96-1.07(m,2H).

[0530] Example 24

[0531] Synthesis route:

[0532]

[0533] Step 1: Preparation of Compound 24

[0534] At 0 °C, sodium hydride (39.46 mg, 986.57 μmol) was added to a DMF (4 mL) solution of compound 2-9 (0.10 g, 246.6-4.5 μmol). After 10 minutes, compound 1-fluoro-2-iodoethane (85.81 mg, 493.28 μmol) was added to the reaction solution and stirred for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (10 mL), extracted with dichloromethane (10 mL * 2), washed with saturated brine (10 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Compound 24 was purified by preparative HPLC (column: YMC Triart 30 * 150 mm * 7 μm; mobile phase: hydrochloric acid aqueous solution and acetonitrile; the proportion of acetonitrile in the mobile phase ranged from 13% to 33%, 10 min). LCMS (ESI) m / z: 404.2 (M+1). 1H NMR (400MHz, DMSO-d6) δ = 8.93 (s, 1H), 8.83 (s, 1H), 8.54 (d, J = 8.4Hz, 1H), 7.84 (d, J = 8.4Hz, 1H), 7.57 (s, 1H), 4.83 (br s,2H),4.80-4.72(m,2H),4.60(s,2H),4.51-4.44(m,1H),4.09-4.01(m,2H),3.20-3.18(m,3H),1.32(br d,J=6.5Hz,2H),1.04(br s,2H).

[0535] Example 25

[0536] Synthesis route:

[0537]

[0538] Step 1: Preparation of Compound 25

[0539] At 0 °C, sodium hydride (1.58 mg, 39.56 μmol) was added to a DMF (2 mL) solution of compound 22-1 (0.02 g, 39.56 mmol). After 10 minutes, iodomethane (39.56 μmol, 2.46 μL) was added to the reaction solution and stirred for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (10 mL), extracted with dichloromethane (10 mL * 2), washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Compound 25 was purified by preparative HPLC (column: Waters Xbridge 150 * 25 mm * 5 μm; mobile phase: 0.05% ammonium bicarbonate aqueous solution and acetonitrile; acetonitrile content in the mobile phase ranged from 29% to 59%, 9 min). LCMS (ESI) m / z: 492.2 (M+1). 1 H NMR (400MHz, DMSO-d6) δ = 8.88 (s, 1H), 8.54 (s, 1H), 8.44 (d, J = 8.8Hz, 1H), 7.9 1(d,J=8.0Hz,1H),6.93(s,1H),4.70(brd,J=1.5Hz,2H),4.52(s,2H),4.48(br dd,J=2.9,5.8Hz,1H),4.42-4.32(m,2H),3.92-3.84(m,1H),3.79-3.65(m,2H) ,3.18(s,3H),1.39(s,3H),1.36-1.31(m,2H),1.28(s,3H),1.26-1.22(m,2H).

[0540] Example 26

[0541] Synthesis route:

[0542]

[0543] Step 1: Preparation of Compound 26

[0544] At room temperature, 0.5 mL of an aqueous solution of sodium hydroxide (15.82 mg, 395.60 μmol) was added to 1 mL of methanol containing compound 22-1 (20.00 mg, 39.56 μmol), and the mixture was stirred at 40 °C for 16 hours. The mixture was filtered, and the filtrate was purified by preparative HPLC (Waters Xbridge 150*25 mm*5 μm column; mobile phase: 0.05% ammonium bicarbonate aqueous solution and acetonitrile; acetonitrile content in the mobile phase ranged from 25% to 55%, 10 min) to give compound 26. LCMS (ESI) m / z: 478.3 (M+1). 1H NMR (400MHz, CD3OD) δppm 8.83 (br s, 1H), 8.51 (br d,J=8.31Hz,1H), 8.37(m,1H), 7.69(m,1H), 6.72(s,1H), 4.56-4.68(m,2H), 4.51(s,2H), 4.36-4.44(m,1H), 4.25-4.34( m,1H), 4.13(dd,J=8.25,6.30Hz,1H), 3.79(dd,J=8.38,6.30Hz,1H), 3.60(dd,J=8.19,5.62Hz,2H), 1.45(s,3H), 1.41(br d,J=6.72Hz,2H),1.37(s,3H),1.01(br s,2H).

[0545] Example 27

[0546]

[0547] Synthesis route:

[0548]

[0549] Step 1: Preparation of Compound 27-1

[0550] Compound 7-7 (50 mg, 127.74 μmol) was dissolved in anhydrous dichloromethane (5 mL) at room temperature, followed by the sequential addition of N,N-di-tert-butyloxycarbonyl-thiourea (42.36 mg, 153.29 μmol), triethylamine (38.78 mg, 383.22 μmol), and copper chloride (20.61 mg, 153.29 μmol). The reaction mixture was stirred at 20 °C for 12 hours. The reaction mixture was then filtered and concentrated under reduced pressure to obtain compound 27-1. LCMS (ESI) m / z: 634.3 (M+1).

[0551] Step 2: Preparation of trifluoroacetate of compound 27

[0552] Compound 27-1 (80 mg, 126.24 μmol) was dissolved in ethanol (3 mL) and water (2 mL) at room temperature. 12 M hydrochloric acid (12 M, 2 mL) was added to the reaction solution, and the mixture was stirred at 80 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase: 0.05% trifluoroacetic acid aqueous solution and acetonitrile; acetonitrile content in the mobile phase ranged from 18% to 48%, 9 min) to obtain the trifluoroacetate of compound 27. LCMS (ESI) m / z: 406.2 (M+1).

[0553] Example 28

[0554]

[0555] Synthesis route:

[0556]

[0557] Step 1: Preparation of compound 28-2

[0558] Compound 28-1 (1 g, 4.55 mmol) was dissolved in DCM (10 mL) at 0 °C, and sodium hydroxide (182.12 mg, 4.55 mmol) was slowly added to water (10 mL). The reaction was carried out at 0-25 °C for 5 hours. The reaction solution was diluted with water (20 mL), extracted with dichloromethane (60 mL), the organic layer was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 28-2. LCMS (ESI) m / z: 184.1 (M+1).

[0559] Step 2: Preparation of compound 28-3

[0560] Compound 28-2 (19.50 mg, 106.45 μmol) and compound 7-7 (50 mg, 127.74 μmol) were dissolved in acetonitrile (1 mL), and then calcium trifluoromethanesulfonate (15.00 mg, 40.67 μmol) was added. The mixture was stirred at 45 °C for 16 h. If the reactants were not completely reacted, the reaction solution was heated to 60 °C and the reaction was continued for another 16 h. The reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: YMC Triart C18 150*25 mm*5 μm; mobile phase: [0.01% hydrochloric acid aqueous solution and acetonitrile; the proportion of acetonitrile in the mobile phase ranged from 16% to 46%, 8.5 min) to obtain compound 28-3. LCMS (ESI) m / z: 575.3 (M+1).

[0561] Step 3: Preparation of trifluoroacetate of compound 28

[0562] At room temperature, sodium hydroxide (6.96 mg, 174.04 μmol) and water (0.5 mL) solution were added to an anhydrous methanol (1 mL) solution of compound 28-3 (20 mg, 34.81 μmol). The mixture was stirred at 25 °C for 2 h. The reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Phenomenex Synergi Polar-RP 100*25 mm*4 μm; mobile phase [0.05% trifluoroacetic acid aqueous solution and acetonitrile; acetonitrile content in the mobile phase from 15% to 45%, 9 min] to give the trifluoroacetate of compound 28. LCMS (ESI) m / z: 547.3 (M+1).

[0563] Example 29

[0564]

[0565] Synthesis route:

[0566]

[0567] Step 1: Preparation of compound 29-2

[0568] Compound 7-7 (50 mg, 127.74 μmol) and compound 29-1 (92.33 mg, 255.48 μmol) were dissolved in methanol (2 mL) and dichloromethane (4 mL) at room temperature. Acetic acid (11.5 mg, 191.61 μmol) was then added, and the mixture was stirred at 40 °C for 1 hour. The mixture was then cooled to 10-20 °C, and pyridineborane (35.61 mg, 383.22 μmol) was added. The mixture was stirred at 40 °C for 16 hours. The reaction solution was concentrated under reduced pressure to give compound 29-2. LCMS (ESI) m / z: 737.4 (M+1).

[0569] Step 2: Preparation of the hydrochloride salt of compound 29

[0570] Compound 29-2 (94 mg, 127.58 μmol) was dissolved in water (2 mL) and methanol (3 mL) at room temperature, and concentrated hydrochloric acid (1.02 g, 10 mmol) was added. The mixture was stirred at 50-55 °C for 16 hours. The reaction solution was purified by preparative HPLC (column: YMCTriart C18150*25 mm*5 μm; mobile phase: 0.01% hydrochloric acid aqueous solution and acetonitrile; the proportion of acetonitrile in the mobile phase ranged from 9% to 29%, 10 min) to obtain the hydrochloride salt of compound 29. LCMS (ESI) m / z: 509.2 (M+1).

[0571] Biological evaluation:

[0572] Experimental Example 1: Detection of Minimum Inhibitory Concentration (MIC)

[0573] 1) Preparation of compound masterbatch

[0574] On the day of the experiment, dissolve the compound in the bottle in 100% DMSO until the stock solution concentration is 0.4 mg / mL. If the stock solution is not used on the same day, store it at -80℃ for later use.

[0575] The stock solution of the compound and the standard antibiotic were serially diluted two-fold using appropriate solvents on 96-well microplates (V bottom) to obtain 100× working solutions (wells 1 to 7). The concentrations of the compound were 400, 200, 100, 50, 25, 12.5, 6.25, and 3.125 μg / mL. 100% DMSO was used as a growth control (well 8). This is the stock solution of the compound.

[0576] 2) Preparation of inoculum

[0577] One day before the experiment, aerobic bacteria (Staphylococcus epidermidis) stored in -80℃ cryovials were streaked onto MHA (Cation-adjusted Mueller-Hinton agar) plates. The plates were then incubated aerobically at 35±2℃ for 18-24 hours.

[0578] Two days before the experiment, anaerobic bacteria (Propionibacterium acnes) from -80℃ cryopreserved tubes were streaked onto MBA (Brucella Agar + 5% (v / v) defibrinated sheep blood + 5 μg / mL Hemin + 1 μg / mL Vitamin K1) plates. The plates were then incubated at 35 ± 2℃ in an anaerobic incubator for 42–48 hours.

[0579] On the day of the experiment, the plates were removed, clones were picked from the plates, and suspended in physiological saline. The turbidity of the bacterial suspension was then adjusted to OD600 = 0.2 using a turbidimeter. This bacterial suspension contained 1.0-2.0 × 10⁻⁶ mg / L. 8 CFU / mL. Then, dilute the turbidity-adjusted bacterial suspension with test medium to the appropriate concentration: ~5 × 10⁻⁶ CFU / mL for aerobic bacteria. 5 CFU / mL, anaerobic bacteria: ~1×10⁻⁶ 6 CFU / mL. This is the inoculum.

[0580] 3) Minimum inhibitory concentration (MIC) detection

[0581] Transfer 2 μl of a 2-fold serially diluted 100× high-concentration working solution from the compound master plate (prepared in step 1) to a round-bottom 96-well plate. Then, add 198 μL of bacterial inoculum (prepared in step 2) to each well to obtain the MIC test plate. Therefore, the final test concentrations of the compounds are 4, 2, 1, 0.5, 0.25, 0.125, 0.06, and 0.03 μg / mL. The total volume per well of the test plate is 200 μL: containing 1% DMSO, ~5×10⁻⁶ mg / mL.5 CFU / mL of aerobic bacteria and ~1×10 6 Anaerobic bacteria at CFU / mL.

[0582] All aerobic bacteria test plates were placed in an aerobic incubator at 35 ± 2℃ for 16-20 hours, and all anaerobic bacteria test plates were placed in an anaerobic incubator at 35 ± 2℃ for 48 hours.

[0583] Note: For anaerobic bacteria, steps 2 (bacterial culture preparation) and 3 (bacterial culture inoculation) are both performed in an anaerobic workstation.

[0584] 4) Read the minimum inhibitory concentration (MIC)

[0585] After incubation, the drug concentration in the wells that completely or significantly inhibit bacterial growth is the minimum inhibitory concentration (MIC) observed by the naked eye on the test plate.

[0586] 5) The experimental results are shown in Table 1.

[0587] Table 1. Results of minimum inhibitory concentration (MIC) tests for the compounds of this invention.

[0588]

[0589]

[0590] Experimental conclusion: The compound of this invention has good in vitro antibacterial activity.

[0591] Experimental Example 2: Pharmacokinetic Evaluation of Compounds

[0592] 1) Experimental materials:

[0593] CD-1 mice (male, Beijing Vital River Laboratory Animal Technology Co., Ltd.).

[0594] 2) Experimental procedures:

[0595] The pharmacokinetic characteristics of the compounds after intravenous and oral administration were tested in rodents using a standard protocol. In the experiment, candidate compounds were prepared into clear solutions and administered to mice via single intravenous and oral injections. The solvent for both intravenous and oral administration was a mixture of 10% DMSO and 90% (20% SBE-β-CD). Four male CD-1 mice were used in this study. Two mice received intravenous administration at a dose of 1 mg / kg, and plasma samples were collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours post-administration. The other two mice received oral gavage at a dose of 2 mg / kg, and plasma samples were collected at 0.25, 0.5, 1, 2, 4, 8, and 24 hours post-administration. Blood samples were placed on ice after collection and centrifuged within one hour (centrifugation conditions: 6000g, 3 minutes, 2-8℃). Plasma samples were stored at -80℃ before analysis. Blood drug concentrations were quantitatively analyzed using LC-MS / MS, and pharmacokinetic parameters, such as peak concentration (C0), were calculated. max ), clearance rate (CL), half-life (T) 1 / 2 ), tissue distribution (Vdss), area under the curve (AUC) 0-last ), bioavailability (F), etc.

[0596] 3) The experimental results are shown in Table 4.

[0597] Table 4. Pharmacokinetic test results of the compounds of this invention.

[0598]

[0599] Experimental conclusion: The compounds of this invention have good pharmacokinetic properties, including good oral bioavailability and oral exposure.

[0600] Experimental Example 3: In vivo pharmacodynamic study of compounds

[0601] 1) Experimental materials

[0602] Experimental strain: Propionibacterium acnes ATCC 6919 (Beijing BioBio Biotechnology Co., Ltd.).

[0603] Laboratory animals: Balb / C mice (Zhejiang Vital River Laboratory Animal Technology Co., Ltd.)

[0604] 2) Experimental methods

[0605] 1. Establishment of an acne model:

[0606] Balb / C mice were grouped and numbered, and acclimatized for one week. On the day of modeling, Propionibacterium acnes was collected, and the OD600 was measured using a microplate reader. BHI medium was diluted to OD600 = 1 (approximately 5 × 10⁻⁶). 8(CFU / mL), centrifuge and discard the supernatant, then dilute with PBS to 1×10⁻⁶. 8 CFU / mL. After modeling begins, mice are anesthetized using a small animal anesthesia machine. The gas volume and isoflurane content of the anesthesia machine are adjusted, and the mice are placed in an anesthesia induction box for anesthesia induction. After no obvious signs of movement are observed, the mice are removed, and their toes are grasped with forceps or hemostatic forceps to perform a toe pinching response test. If there is no pain response to withdrawal, surgery can be performed. The anesthetized mice are fixed on a mouse board, and the gas volume or isoflurane content is adjusted to maintain anesthesia (isoflurane anesthesia induction content 3%–4%, anesthesia maintenance content 1%–2%, air flow rate: 300–500 mL / min). Blank control group: 20 μL of PBS is injected intradermally into the left ear of each mouse. Model group and test drug group: 20 μL of Propionibacterium acnes PBS suspension is injected intradermally into the left ear of each mouse. 4, 20, 28, and 44 hours after the injection of Propionibacterium acnes, mice in each group are administered the drug according to the administration method and dosage described in Table 5.

[0607] Forty-eight hours after injection of Propionibacterium acnes, the ears of mice in each group were photographed at the experimental endpoint. The thickness of the left auricle was measured with calipers. The left ear was taken for tissue homogenization to detect the amount of bacteria in the ear and the levels of IL-6 and IL-8 (mice do not express IL-8, so the functional homologue CXCL1 / KC was detected).

[0608] 2. Preparation of the medicine:

[0609] The test drugs are compounds 6, 7 and 12, all of which are hydrochloride salts. In this experiment, compounds 6, 7 and 12 refer to the corresponding hydrochloride salt drugs. The solvent is 40% DMSO + 60% glycerol. The drugs are added to DMSO, heated to 100°C in a water bath, and then glycerol is added to obtain a bright yellow viscous solution.

[0610] 3. For administration methods and dosages, please refer to Table 5 below.

[0611] Table 5. Administration Methods and Dosage Table

[0612]

[0613] Administration area: Injection of Propionibacterium acnes into the left ear.

[0614] Administration time: 4, 20, 28, and 44 hours after injection of Propionibacterium acnes, for a total of 4 times.

[0615] 4. Modeling method:

[0616] Balb / C mice in groups 6, 7, 12, and the model group were anesthetized using a small animal anesthesia machine and isoflurane. After maintaining anesthesia on a plateau, mice in the model group and the test drug group were intradermally injected with 1×10⁻⁶ compound solution into the left ear using a 1 mL sterile syringe.8 20 μL of CFU / mL Propionibacterium acnes PBS suspension was injected into the blank control group, which was injected with an equal volume of 1×PBS.

[0617] 5. Testing indicators:

[0618] Gross photography: The mouse ears were photographed 48 hours after injection of Propionibacterium acnes.

[0619] Ear thickness measurement: 48 hours after injection of Propionibacterium acnes, the ear thickness of mice was measured using calipers.

[0620] Ear-borne bacterial count detection: Forty-eight hours after injection of Propionibacterium acnes, the left ear of mice was homogenized with 1 mL PBS. Three 200 μL aliquots of the homogenate were taken, and each aliquot was diluted with 800 μL PBS. 100 μL of the diluted homogenate was then diluted 10-fold, and the above operation was repeated for a total of 5 dilutions. The homogenate of each dilution was spread on BHI solid medium and incubated anaerobically at 37°C for 48 hours. The colony count was then calculated.

[0621] Mouse BCA protein assay: Forty-eight hours after injection of *Propionibacterium acnes*, a BCA protein concentration assay kit was used. The left ear of each mouse was homogenized with 1 mL of PBS, and the dilution factor equivalent to a 1:9 homogenate was calculated. The homogenate was centrifuged at 1500g for 10 minutes. Each sample was diluted 10-fold with PBS. 20 μL of sample was added to each well of a 96-well plate. 200 μL of BCA working solution was added to each well, and the plate was incubated at 37°C for 15-30 minutes. The A562 level was measured using a microplate reader, a standard curve was plotted, and the BCA concentration was calculated. This yielded the levels of IL-6 and CXCL1.

[0622] 3) The experimental results are shown in Tables 6, 7 and 8.

[0623] Table 6. Effects of the test drugs on ear thickness in acne model mice.

[0624]

[0625] Forty-eight hours after injection of Propionibacterium acnes, the ear thickness of mice in each group was measured using digital calipers. Results are expressed as Mean ± SD. * p<0.05, ** p<0.01, *** p<0.001 compared with the blank control group, # p<0.05, ## p<0.01, ### p<0.001 compared with the model group.

[0626] Table 7. Effects of the test drugs on ear bacterial load in acne model mice.

[0627]

[0628] Forty-eight hours after injection of Propionibacterium acnes, tissue was taken from the left ear for bacterial load testing. Results are expressed as Mean ± SD. * p<0.05, ** p<0.01, *** p<0.001 compared with the model group.

[0629] The left ear of mice in the blank control group showed no change; the left ear of mice in the acne model group showed obvious redness and swelling; after treatment with compound 6, compound 7 and compound 12, the redness and swelling of the left ear of mice were reduced to varying degrees, and the condition improved.

[0630] As shown in Table 6, compared with the blank control group mice, the thickness of the left auricle of the acne model group mice was significantly increased (p<0.001); after treatment with compound 6, compound 7 and compound 12, the thickness of the left auricle of the mice was significantly reduced (p<0.05), among which compound 7 treatment showed the most significant reduction in the thickness of the left auricle of the mice.

[0631] As shown in Table 7, the bacterial load in the left ear of mice was significantly reduced after treatment with compounds 6, 7 and 12 (p<0.001); among them, the bacterial load in the left ear of mice decreased the most after treatment with compound 7.

[0632] Table 8. Effects of the test drugs on IL-6 and CXCL1 levels in acne model mice.

[0633]

[0634] Forty-eight hours after injection of Propionibacterium acnes, the ear thickness of mice in each group was measured using digital calipers. Results are expressed as Mean ± SD. * p<0.05, ** p<0.01, *** p<0.001 compared with the blank control group, # p<0.05, ## p<0.01, ### p<0.001 compared with the model group.

[0635] As shown in Table 8, compared with the blank control group, the levels of IL-6 and CXCL1 in the left ear tissue of mice in the acne model group were significantly increased (p<0.001); after treatment with compound 6, compound 7 and compound 12, the levels of IL-6 and CXCL1 in the left ear tissue of mice were significantly decreased (p<0.001); among them, compound 7 showed the best efficacy.

[0636] 4) Experimental Conclusions

[0637] This study investigated the effects of different drugs on acne-prone mouse models by establishing acne models and immunosuppressed mouse acne models. The results demonstrated that the tested drugs could improve acne-prone mouse symptoms and reduce the levels of IL-6 and CXCL-1 in lesion tissues. Key findings included: compounds 6, 7, and 12 all improved the thickness of the ear lesions in acne-prone mice, alleviating redness and swelling; reduced the bacterial load in the ear lesions, inhibiting the growth of *Propionibacterium acnes*; and decreased the levels of the inflammatory factors IL-6 and CXCL1 in the ear lesions of acne-prone mice.

Claims

1. A compound, stereoisomer or pharmaceutically acceptable salt thereof of formula (II), wherein, T1 is selected from N and CR1; T2 is selected from N and CR2; X is selected from -C(R7R8)-; Y is selected from -0- and -C(R7R8)-; Z is selected from -C(R7R8)-; or, -Y-Z- is selected from -C(R9)=C(R9)-; R1 is selected from H; R2 is selected from H; R5 and R6 are each independently selected from H; each R7, R8 and R9 are each independently selected from H. ， 2. The compound, stereoisomer or pharmaceutically acceptable salt thereof of claim 1, wherein the compound has a structure of formula (II-2): ###0002### (II-2) Y, Z, T1, T2 and R3 are as defined in claim 1. is selected from a single bond and a double bond; 3. A compound, stereoisomer or pharmaceutically acceptable salt thereof of formula (III), ###0003### (III) 4. The compound, stereoisomer or pharmaceutically acceptable salt thereof of claim 3, wherein the compound is: ###0004### 5. Use of the compound, stereoisomer or pharmaceutically acceptable salt thereof of any one of claims 1 to 4 in the manufacture of an antibacterial, anti-inflammatory medicament.

6. Use of the compound, stereoisomer or pharmaceutically acceptable salt thereof of any one of claims 1 to 4 in the manufacture of a medicament for the treatment of acne. ​ ​ ​ ​ R3is selected from H, =0, -NH2, -NHC(=NH)NH2, -CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, -NHCH2CH2CH3, -NHCH2CH2OH, -NHCH2CH2NH2, -OCH3, , , , , , and ; or R1and R3together with the atoms to which they are attached form ; R4is selected from ; ​ ​ ​ ， wherein ​ ​ 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 ​ 、 、 、 、 、 。 ​ ​

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