A nitrogen-containing heterocyclic compound, a preparation method therefor, and an application thereof

Abstract

The present application provides a nitrogen-containing heterocyclic compound or a pharmaceutically acceptable salt, prodrug, deuterated compound, hydrate, solvate, enantiomer, diastereomer or racemate thereof, a preparation method and use thereof. The compound can be used for preparing a fast-acting antidepressant

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, and particularly relates to a compound exerting a rapid anti-depression effect and application thereof. BACKGROUND

[0002] Depression is a common mental disorder with marked and persistent low mood as the main clinical feature. It is characterized by mood depression that is not commensurate with the situation, and emotional depression can range from gloom to despair, self-deprecating depression, and even pessimism and world-weariness, with suicidal attempts or behaviors. In severe cases, psychotic symptoms such as hallucinations and delusions may occur. Globally, more than 350 million people suffer from depression, with an increase of 18% in the past decade. As of 2017, there were more than 54 million depression patients in China. The recurrence rate of major depressive disorder is as high as 50%-85%, and the suicide rate is 4.0%-10.6%. Depression has become the most common mental illness.

[0003] Disorders of neurotransmitters such as norepinephrine, dopamine, and serotonin (5-HT) in the brain are considered to be the main biological factors leading to depression. The main means of treating depression relies on the use of antidepressants. Antidepressants are mainly divided into the following four categories according to their effects and mechanisms: 1) monoamine oxidase inhibitors: such as isopropyl hydrazine and isocarboxazid. Due to the presence of ingredients with relatively large side effects, these drugs have been basically discontinued; 2) tricyclic antidepressants: this type of drug has a good effect on relieving depression, but has a greater adverse effect on patients with other diseases. This drug includes clomipramine and imipramine hydrochloride; 3) selective 5-HT reuptake inhibitors: this drug can compensate for the symptoms of 5-HT deficiency in depression patients. The main drugs are fluoxetine, paroxetine, sertraline, citalopram, and fluvoxamine; 4) 5-HT and norepinephrine reuptake inhibitors: this type of drug has a dual antidepressant mechanism and relatively high safety. Examples include venlafaxine and duloxetine. Classes 3 and 4 are the main drugs, and classes 1 and 2 have been basically discontinued.

[0004] Since the advent of serotonin reuptake inhibitors (SSRi), there have been dozens of generations of iterative upgrades. Currently, there are more than a hundred in research projects, mainly focusing on SSRi, selective serotonin and norepinephrine reuptake inhibitors (SNRI), norepinephrine and specific serotonin antidepressants (NaSSA), 5-HT receptor antagonists and reuptake inhibitors (SARIs), NMDA receptor antagonists, etc. The antidepressant market is also very concentrated. To date, there have been more than 30 SSRi varieties in global application, and the top 10 are escitalopram, sertraline, venlafaxine, paroxetine, duloxetine, flupentixol + mirtazapine, fluoxetine, citalopram, and fluvoxamine, with a combined market share of over 90%.

[0005] However, the currently used antidepressants in clinic have serious limitations, mainly slow onset, lag, and need 3-4 weeks of continuous medication to take effect, and many patients give up treatment because they cannot achieve the desired effect; for patients with suicidal tendencies, their lives may not be saved in time. The (S) enantiomer of ketamine (Esketamine) in the form of nasal spray was listed in the United States in 2019. It can quickly (within a few hours) relieve depressive symptoms and can last for 3-4 days. Although esketamine can take effect quickly, it has serious side effects, including severe drowsiness, dissociative hallucinations, potential addiction, etc. FDA also gives a black box warning, and esketamine must be used with other oral antidepressants for refractory depression in adults. These deficiencies limit the clinical application of such drugs. Therefore, there is an urgent need for new antidepressants with fast onset, good efficacy and oral administration in clinic. SUMMARY

[0006] The purposes of the present application include providing a nitrogen-containing heterocyclic compound, and a pharmaceutically acceptable salt, prodrug, deuterium compound, hydrate, solvate, enantiomer, diastereoisomer or racemate thereof;

[0007] Another purpose of the present application includes providing a pharmaceutical composition comprising a therapeutically effective amount of any of the above compounds, or a pharmaceutically acceptable salt, prodrug, deuterium compound, hydrate, solvate, enantiomer, diastereoisomer or racemate thereof, and a pharmaceutically acceptable excipient;

[0008] Another purpose of the present application includes providing the use of the above compound of general formula or a pharmaceutically acceptable salt, prodrug, deuterium compound, hydrate, solvate, enantiomer, diastereoisomer or racemate thereof, or the above pharmaceutical composition in the preparation of a medicament for preventing and / or treating depression.

[0009] Another purpose of the present application includes providing a preparation method of the above compound of general formula.

[0010] To achieve the above purposes, the technical solutions adopted by the present application include:

[0011] The present application provides a compound represented by general formula (IA) or (IB) or a pharmaceutically acceptable salt, prodrug, deuterium compound, hydrate, solvate, enantiomer, diastereoisomer or racemate thereof:

[0012]

[0013] Wherein:

[0014] X is a bond or CHR5;

[0015] Z1and Z2are independently selected from: CHR6, NR7, oxygen or carbonyl, and Z1and Z2are not simultaneously NR7or oxygen;

[0016] Z3is CHR6or carbonyl;

[0017] Z4, Z5and Z6are independently selected from CR6, N;

[0018] n is an integer from 0 to 4; R1is hydrogen, R6-(CO)-, R6-(SO)-, R6-(SO2)-, R6-O(CO)-, R6R7-N(CO)-, or C1-C 15 chain alkyl, C1-C 15 hetero chain alkyl, C3-C 15 cycloalkyl, C4-C 15 heterocycloalkyl, C5-C 15 aryl, C5-C 15 heteroaryl; R2is optionally selected from carboxyl, C1-C 15 chain alkoxycarbonyl, C1-C 15 hetero chain alkoxycarbonyl, C3-C 15 cycloalkoxycarbonyl, C4-C 15 heterocycloalkoxycarbonyl, C5-C 15 aryl, C5-C 15 heteroaryl;

[0019] R3, R4are each independently optionally selected from hydrogen or C1-C 15 chain alkyl, C1-C 15 hetero chain alkyl, C3-C 15 cycloalkyl, C4-C 15 heterocycloalkyl, C5-C 15 aryl, C5-C 15 heteroaryl;

[0020] R5is optionally selected from hydrogen, C1-C5alkyl, C3-C7cycloalkyl, C4-C7heterocycloalkyl;

[0021] R6is selected from hydrogen, halogen, -CN, -NO2, -OR7, -NR7R8, -SR7, -COR7, -SOR7, -SO2R7, -NR7COR8, -CONR7R8, -OCOR7, -COOR7, -OCOOR7, -OCONR7R8, -NR7CONR8R9, -NR7COOR8, -NR7SO2R8, -SO2NR7R8, -OSO2R7, -SO3R7, a linear alkyl, a hetero-linear alkyl, a cycloalkyl, a hetero-cycloalkyl, an aryl, or a heteroaryl; wherein each linear alkyl, hetero-linear alkyl, cycloalkyl, hetero-cycloalkyl, aryl, or heteroaryl is independently substituted with 0 to 8 halogen, cyano, hydroxyl, thiol, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, alkyl, or haloalkyl; 10 substituted;

[0022] R7, R8, R9are each independently selected from hydrogen, a linear alkyl, a hetero-linear alkyl, a cycloalkyl, a hetero-cycloalkyl, an aryl, or a heteroaryl, wherein each linear alkyl, hetero-linear alkyl, cycloalkyl, hetero-cycloalkyl, aryl, or heteroaryl is independently substituted with 0 to 8 halogen, cyano, hydroxyl, thiol, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, alkyl, or haloalkyl;

[0023] R 10 substituted with 0 to 8 halogen, cyano, hydroxyl, thiol, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, alkyl, or haloalkyl;

[0024] wherein the compounds of general formula (IA) do not include the compounds of general formula (IB) do not include

[0025] The present application also provides a compound of formula (Ia) or (Ib) or a pharmaceutically acceptable salt, prodrug, deuterated analog, hydrate, solvate, enantiomer, diastereomer, or racemate thereof:

[0026]

[0027] wherein:

[0028] X is a bond or CHR5;

[0029] Z1and Z2are independently selected from: CHR6, NR7, oxygen, or carbonyl, and Z1and Z2are not simultaneously NR7or oxygen;

[0030] Z3 is CHR6or carbonyl;

[0031] Z4, Z5and Z6are independently selected from CR6or N;

[0032] n is an integer from 0 to 4;

[0033] R1is selected from hydrogen, R6-(CO)-, R6-(SO)-, R6-(SO2)-, R6-O(CO)-, R6R7N(CO)-, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 membered heteroaryl, wherein C1-C 15 alkyl, wherein the heteroatom is selected from one or more of O, S, N, said C1-C 15 alkyl is optionally substituted with 0-8 R6;

[0034] R2is selected from -(CO)O-(CH2) m R6, wherein m is an integer from 0 to 4, or R2is selected from C6-C 15 aryl or 5-15 membered heteroaryl, wherein C6-C 15 aryl or 5-15 membered heteroaryl, wherein C6-C

[0035] R3, R4are each independently selected from hydrogen, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 membered heteroaryl, wherein C1-C 15 alkyl, wherein the heteroatom is selected from one or more of O, S, N, said C1-C 15chain alkyl is optionally substituted with 0-8 R6;

[0036] R5is selected from hydrogen, C1-C5alkyl, C3-C7cycloalkyl, or 4-7 membered

[0037] R6is selected from hydrogen, halogen, -CN, -NO2, -OR7, -NR7R8, -SR7, -(CO)-R7, -(SO)-R7, -(SO2)-R7, -NR7-(CO)-R8, -(CO)-NR7R8, -O(CO)-R7, -(CO)OR7, -O(CO)OR7, -O(CO)-NR7R8, -NR7-(CO)-NR8R9, -NR7-(CO)OR8, -NR7-(SO2)-R8, -(SO2)-NR7R8, -O(SO2)-R7, -(SO2)OR7, C1-C 15 alkyl, C1-C 15 alkyl, C1-C 15 alkyl, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl, or 5-15 membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkyl, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 10 aryl, or 5-15 membered heteroaryl, optionally substituted with 0-8 R 15 chain alkyl, wherein the heteroatom is selected from one or more of O, S, N, said C1-C 15 chain alkyl is optionally substituted with 0-8 R 10 ;

[0038] R7, R8, R9are each independently selected from hydrogen, C1-C 15 alkyl, C1-C 15 alkyl, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl, or 5-15 membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkyl, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl, or 5-15 membered heteroaryl, optionally substituted with 0-8 halogen, cyano, hydroxyl, thiol, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, alkyl, or haloalkyl; or, R7, R8, R9are each independently selected from C1-C 15Chain alkyl group, wherein the heteroatom is selected from one or more of O, S, and N, wherein the C1-C containing the heteroatom 15 The chain alkyl group is optionally substituted with 0 to 8 halogen, cyano, hydroxyl, mercapto, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate, sulfonamide, alkyl or haloalkyl groups.

[0039] R 10 Selected from hydrogen, halogen, cyano, hydroxyl, mercapto, ether, nitro, amino, carboxyl, sulfonic acid, ester, amide, sulfonate, sulfonamide, C1-C 15 Alkyl, C1-C 15 Alkoxy, C3-C 15 Cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 heteroaryl, wherein C1-C 15 Alkyl, C1-C 15 Alkoxy, C3-C 15 Cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 The aryl or 5-15 heteroaryl groups are optionally substituted with 0 to 8 halogens, cyano, hydroxyl, mercapto, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate, sulfonamide, alkyl, or haloalkyl groups.

[0040] Formula (Ia) does not contain the following compounds:

[0041]

[0042]

[0043]

[0044] Formula (Ib) does not contain the following compounds:

[0045]

[0046] In some implementations, wherein,

[0047] X is the key;

[0048] Z1 and Z2 are independently selected from: CHR6, NR7, oxygen or carbonyl, and Z1 and Z2 are not simultaneously NR7 or oxygen; Z3 is CHR6 or carbonyl.

[0049] Z4, Z5 and Z6 are independently selected from CR6 or N;

[0050] n is an integer from 0 to 4;

[0051] R1 is selected from hydrogen, R6-(CO)-, R6-(SO)-, R6-(SO2)-, R6-O(CO)-, R6R7N(CO)-, C1-C 15 Alkyl, C1-C 15 Alkoxy, C3-C 15 Cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 heteroaryl, wherein C1-C 15 Alkyl, C1-C 15 Alkoxy, C3-C 15 Cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 The aryl or 5-15 heteroaryl groups are optionally substituted by 0 to 8 R6 groups;

[0052] R2 is selected from -(CO)O-(CH2) m R6, where m is an integer from 0 to 4, or R2 is selected from C6-C 15 aryl or 5-15 heteroaryl, of which C6-C 15 The aryl or 5-15 heteroaryl groups are optionally substituted by 0 to 8 R6 groups;

[0053] R3 and R4 are each independently selected from hydrogen or C1-C2. 15 Alkyl, wherein C1-C 15 Alkyl groups are optionally substituted with 0 to 8 R6 groups; R3 and R4 are not both hydrogen;

[0054] R6 is selected from hydrogen, halogen, -CN, -NO2, -OR7, -NR7R8, -SR7, -(CO)-R7, -(SO)-R7, -(SO2)-R7, -NR7-(CO)-R8, -(CO)-NR7R8, -O(CO)-R7, -(CO)OR 7. -O(CO)OR7, -O(CO)-NR7R8, -NR7-(CO)-NR8R9, -NR7-(CO)OR8, -NR7-(SO2)-R8, -(SO2)-NR7R8, -O(SO2)-R7, -(SO2)OR7, C1-C 15 Alkyl, C1-C 15 Alkoxy, C3-C 15 Cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 heteroaryl, wherein C1-C 15 Alkyl, C1-C 15 Alkoxy, C3-C 15 Cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 Aryl or 5-15 quinone heteroaryl groups are optionally surrounded by 0-8 R groups. 10 replace;

[0055] R7, R8, R9are each independently selected from hydrogen, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, optionally substituted with 0 to 8 halogen, cyano, hydroxyl, thiol, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, alkyl or haloalkyl;

[0056] R 10 selected from hydrogen, halogen, cyano, hydroxyl, thiol, ether, nitro, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, optionally substituted with 0 to 8 halogen, cyano, hydroxyl, thiol, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, alkyl or haloalkyl.

[0057] In some embodiments, wherein,

[0058] the compound of Formula (Ia) is a compound of Formula (Ia-1), and the compound of Formula (Ib) is a compound of Formula (Ib-1)

[0059]

[0060] wherein R1, R2, Z1, Z2, Z3, Z4, Z5, Z6and n are as defined above.

[0061] In some embodiments, wherein,

[0062] the compound of Formula (Ia) is a compound of Formula (Ia-1’),

[0063]

[0064] wherein R1, R2, R3, R4 are as defined above.

[0065] In some embodiments, wherein,

[0066] The compound of formula (Ia) is a compound of formula (Ia-2) as follows,

[0067]

[0068] wherein R1, R2 are as defined above.

[0069] In some embodiments, wherein,

[0070] The compound of formula (Ia) is a compound of formula (Ia-3) as follows,

[0071]

[0072] wherein R2 is as defined above.

[0073] In some embodiments, wherein,

[0074] R2 is selected from -(CO)O-(CH2) m R6, wherein m is an integer from 0 to 4.

[0075] In some embodiments, wherein,

[0076] R2 is selected from C6-C 15 aryl or 5-15 membered heteroaryl, wherein C6-C 15 aryl or 5-15 membered heteroaryl, optionally substituted with 0-8 R6.

[0077] In some embodiments, wherein,

[0078] R2 is selected from:

[0079]

[0080] In some embodiments, wherein,

[0081] X is a bond;

[0082] Z1 and Z2 are independently selected from: CHR6, NR7, oxygen or carbonyl, and Z1 and Z2 are not simultaneously NR7 or oxygen;

[0083] Z3 is CHR6 or carbonyl;

[0084] Z4, Z5 and Z6 are independently selected from CR6 or N;

[0085] n is an integer from 0 to 4.

[0086] R1is selected from hydrogen, R6-(CO)-, R6-(SO)-, R6-(SO2)-, R6-O(CO)-, R6R7N(CO)-, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 membered heteroaryl optionally substituted with 0-8 R6;

[0087] R2is selected from C6-C 15 aryl or 5-15 membered heteroaryl, wherein C6-C 15 aryl or 5-15 membered heteroaryl optionally substituted with 0-8 R6;

[0088] R3, R4are each independently selected from hydrogen, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15 aryl or 5-15 membered heteroaryl optionally substituted with 0-8 R6; R3and R4are not hydrogen simultaneously;

[0089] R6is selected from hydrogen, halogen, -CN, -NO2, -OR7, -NR7R8, -SR7, -(CO)-R7, -(SO)-R7, -(SO2)-R7, -NR7-(CO)-R8, -(CO)-NR7R8, -O(CO)-R7, -(CO)OR7, -O(CO)OR7, -O(CO)-NR7R8, -NR7-(CO)-NR8R9, -NR7-(CO)OR8, -NR7-(SO2)-R8, -(SO2)-NR7R8, -O(SO2)-R7, -(SO2)OR7, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3-15 membered heterocycloalkyl, C6-C 15aryl or 5- to 15-membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, optionally substituted with 0 to 8 R 10 substituents;

[0090] R7, R8, R9are each independently selected from the group consisting of hydrogen, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, optionally substituted with 0 to 8 halogen, cyano, hydroxyl, thiol, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, alkyl, or haloalkyl;

[0091] R 10 is selected from the group consisting of hydrogen, halogen, cyano, hydroxyl, thiol, ether, nitro, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, wherein C1-C 15 alkyl, C1-C 15 alkoxy, C3-C 15 cycloalkyl, 3- to 15-membered heterocycloalkyl, C6-C 15 aryl or 5- to 15-membered heteroaryl, optionally substituted with 0 to 8 halogen, cyano, hydroxyl, thiol, ether, nitro, alkoxy, amino, carboxyl, sulfonic acid, ester, amide, sulfonate ester, sulfonamide, alkyl, or haloalkyl.

[0092] In some embodiments, wherein,

[0093] the compound of Formula (Ia) is a compound of Formula (Ia-4) as shown below, and the compound of Formula (Ib) is a compound of Formula (Ib-2) as shown below

[0094]

[0095] wherein R1, R2, R3, Z1, Z2, Z3, Z4, Z5, Z6, and n are as defined above.

[0096] In some embodiments, wherein,

[0097] The compound of Formula (Ia) is a compound of Formula (Ia-5)

[0098]

[0099] wherein R1, R2, R3 are as defined above.

[0100] In some embodiments, wherein,

[0101] The compound of Formula (Ia) is a compound of Formula (Ia-6)

[0102] wherein R2, R3 are as defined above.

[0103] In some embodiments, wherein,

[0104] The compound is selected from any one of the following compounds:

[0105]

[0106]

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113] In some embodiments, wherein, the compound is selected from any one of the following compounds:

[0114]

[0115]

[0116]

[0117]

[0118] The present application also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound described herein, a pharmaceutically acceptable salt, prodrug, deuterated analog, hydrate, solvate, enantiomer, diastereomer, or racemate thereof, and a pharmaceutically acceptable excipient or auxiliary ingredient.

[0119] The present application also provides a use of a compound described herein or a pharmaceutically acceptable salt, prodrug, deuterated analog, hydrate, solvate, enantiomer, diastereomer, or racemate thereof, or the pharmaceutical composition in the preparation of a medicament for preventing and / or treating depression.

[0120] In some embodiments, the medicament in the use described above is a fast-acting medicament.

[0121] The hydrogen described herein can be in any isotopic form, including 1 H (protium), 2 H (deuterium or tritium). 3 H (protium),

[0122] Definitions

[0123] Unless otherwise indicated, the terms used in the specification and claims have the following meanings.

[0124] The carbon, hydrogen, oxygen, sulfur, nitrogen or F, Cl, Br, I involved in the groups and compounds described in the present application include their isotopic forms, and the carbon, hydrogen, oxygen, sulfur or nitrogen involved in the groups and compounds described in the present application are optionally further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include 12 C, 13 C, and 14 C, the isotopes of hydrogen include protium (H), deuterium (D, also called heavy hydrogen), tritium (T, also called super-heavy hydrogen), the isotopes of oxygen include 16 O, 17 O, and 18 O, the isotopes of sulfur include 32 S, 33 S, 34 S, and 36 S, the isotopes of nitrogen include 14 N, and 15 N, the isotopes of fluorine include 17 F, and 19 F, the isotopes of chlorine include 35 Cl, and 37 Cl, the isotopes of bromine include 79 Br, and 81 Br.

[0125] "Alkyl" means a straight or branched chain saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms, preferably an alkyl group of 1 to 15 carbon atoms, more preferably an alkyl group of 1 to 8 carbon atoms, even more preferably an alkyl group of 1, 2, 3, 4, 5, or 6 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, neo-butyl, t-butyl, n-pentyl, i-pentyl, neo-pentyl, n-hexyl, and various branched isomers thereof, and the like. The alkyl group can be optionally further substituted with one or more substituents.

[0126] "Aryloxy" means an aryl group, as defined above, appended to the parent molecular moiety through an oxygen atom. Non-limiting examples of aryloxy groups include phenyloxy, naphthyloxy, and the like. The aryloxy group can be optionally further substituted with one or more substituents.

[0127] "Cycloalkyl" means a saturated or partially unsaturated monocyclic or polycyclic non-aromatic ring substituent, the cycloalkyl group containing 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, more preferably 3 to 8 carbon atoms (e.g., 3, 4, 5, 6, 7, 8 carbon atoms). Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spiro, fused, and bridged ring cycloalkyl groups. The cycloalkyl group can be optionally further substituted with one or more substituents.

[0128] "Heteroaralkyl" refers to an alkyl group substituted with a heteroaromatic group. The heteroaromatic group can be a 5- to 8-membered (e.g., 5-, 6-, 7-, or 8-membered) monocyclic, 8- to 12-membered (e.g., 8-, 9-, 10-, 11-, or 12-membered) bicyclic, or 10- to 15-membered (e.g., 10-, 11-, 12-, 13-, 14-, or 15-membered) tricyclic ring system, which can be bridged or spiro, and contains 1 to 6 (e.g., 1, 2, 3, 4, 5, or 6) heteroatoms selected from N, O, and S. Non-limiting examples of heteroaralkyl groups include pyridylalkyl, pyrimidinylalkyl, pyrazolylalkyl, imidazolylalkyl, thienylalkyl, furanylalkyl, and the like. The heteroaralkyl group can be optionally further substituted with one or more substituents.

[0129] "Aryl" refers to a substituted or unsubstituted aromatic ring, which can be a 5- to 8-membered (e.g., 5-, 6-, 7-, or 8-membered) monocyclic, 6- to 12-membered (e.g., 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered) bicyclic, or 10- to 15-membered (e.g., 10-, 11-, 12-, 13-, 14-, or 15-membered) tricyclic ring system, which can be bridged or spiro, and non-limiting examples include phenyl, naphthyl, and the like. The aryl group can be optionally further substituted with one or more substituents.

[0130] "Heteroaryl" refers to an aromatic ring group having a conjugated planar ring system and containing heteroatoms, which can be a 5- to 8-membered (e.g., 5-, 6-, 7-, or 8-membered) monocyclic, 8- to 12-membered (e.g., 8-, 9-, 10-, 11-, or 12-membered) bicyclic, or 10- to 15-membered (e.g., 10-, 11-, 12-, 13-, 14-, or 15-membered) tricyclic ring system, and contains 1 to 6 (e.g., 1, 2, 3, 4, 5, or 6) heteroatoms selected from N, O, and S. Non-limiting examples of heteroaryl groups include oxazolyl, triazolyl, pyridyl, furanyl, thienyl, pyranyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinyl, benzimidazolyl, benzopyridyl, pyrrolopyridyl, and the like. The heteroaryl group can be optionally further substituted with one or more substituents.

[0131] "Pharmaceutically acceptable salt" means a salt of a compound of the present application which retains the biological effectiveness and properties of the free acids or bases and which is obtained by reaction with a non-toxic inorganic or organic base, for the free acids, or a non-toxic inorganic or organic acid, for the free bases.

[0132] "Pharmaceutical composition" means a mixture of one or more of the compounds of the present application, pharmaceutically acceptable salts or prodrugs thereof, and other chemical components, such as pharmaceutically acceptable carriers, excipients, and / or one or more other therapeutic or prophylactic agents.

[0133] "Optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, and thus such phrases include instances where the event or circumstance occurs and instances where it does not. For example, "heterocyclyl optionally substituted with alkyl" means that the alkyl group can or can not be present, and such phrases include instances where the heterocyclyl is substituted with alkyl and instances where the heterocyclyl is not substituted with alkyl.

[0134] In this document, when when a chemical bond is represented by when a chemical bond is represented by

[0135] The beneficial technical effects achieved are:

[0136] (1) The compounds of the present application have good in vivo rapid antidepressant activity.

[0137] (2) The compounds of the present application have good pharmacokinetic properties, such as good AUC, bioavailability, etc. parameters, compared to the compounds disclosed in the prior art. BRIEF DESCRIPTION OF DRAWINGS

[0138] Figure 1 , Figure 2 shows the results of the tail suspension test in Experimental Example 5.

[0139] Figure 3 shows the results of the forced swimming test in Experimental Example 6.

[0140] Figure 4 shows the results of the tail suspension test in Experimental Example 6. DETAILED DESCRIPTION

[0141] The various starting materials / reagents / instruments used in the examples of the present application are, unless otherwise specified, conventional commercially available products.

[0142] In the present application, 1The instrument for HNMR detection is Bruker Advance 400MHz spectrometer; the instrument for MS-ESI detection of the application is Agilent LCMS1260-6120.

[0143] Preparation of compound A-1-7 in Preparation Example 1

[0144]

[0145] First step

[0146] L-valine methyl ester hydrochloride (779 mg, 4.65 mmol), 4-dimethylaminopyridine (1.3 g, 10.80 mmol) were dissolved in N,N-dimethylformamide (10 mL), cooled to 0 °C, and compound A-1-7a (600 mg, 5.40 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.1 g, 5.94 mmol) were added. The reaction was carried out at room temperature for 16 hours, and TLC showed that the reaction was completed. Water (50 mL) was added for dilution, and ethyl acetate was extracted three times (50 mL x 3). The organic phase was combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was purified by preparative HPLC to obtain compound A-1-7 (580 mg, 2.59 mmol, yield 47.89%). MS-ESI calculated value [M+H] + : 225.12; found: 224.9.

[0147] Preparation of compound A-1-1 in Preparation Example 2

[0148]

[0149] First step

[0150] L-valine methyl ester hydrochloride (686 mg, 4.09 mmol), 4-dimethylaminopyridine (1.2 g, 9.52 mmol) were dissolved in N,N-dimethylformamide (10 mL), cooled to 0 °C, and compound A-1-1a (600 mg, 4.76 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.0 g, 5.32 mmol) were added. The reaction was carried out at room temperature for 16 hours, and TLC showed that the reaction was completed. Water (50 mL) was added for dilution, and ethyl acetate was extracted three times (50 mL x 3). The organic phase was combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was purified by preparative HPLC to obtain compound A-1-1 (750 mg, 3.13 mmol, yield 65.8%). MS-ESI calculated value [M+H] + : 240.1; found: 240.0.

[0151] Preparation of compound A-2-1 of Preparation Example 3

[0152]

[0153] First step

[0154] A-2-1a (5.00 g, 49.45 mmol), di-tert-butyl dicarbonate (12.95 g, 59.34 mmol) and ethanol (60 mL) were added into a 200 mL dry flask, after stirring well, sodium hydroxide (1.98 g, 49.45 mmol) was dissolved in water (25 mL) and added into the reaction system, and the reaction was carried out at room temperature for 16 hours. After the reaction was completed by monitoring by LCMS, water (60 mL) was added for dilution, and 1M dilute hydrochloric acid was used to adjust the pH to 3, and ethyl acetate (150 mL x 3) was used for extraction. The organic phase was combined, washed with saturated brine in turn, dried over anhydrous sodium sulfate, and concentrated to obtain intermediate A-2-1b (8.60 g, 42.74 mmol, yield 86%).

[0155] Second step

[0156] L-valine methyl ester hydrochloride (7.52 g, 44.86 mmol), 4-dimethylaminopyridine (10.45 g, 85.45 mmol) and N,N-dimethylformamide (80 mL) were added into a 200 mL dry flask, and compound A-2-1b (8.60 g, 42.74 mmol) was added at 0°C, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (9.83 g, 51.29 mmol), and the reaction was carried out at room temperature for 16 hours. After the reaction was completed by monitoring by LCMS, water (100 mL) was added for dilution, and ethyl acetate (200 mL x 3) was used for extraction. The organic phase was combined, and washed with water (500 mL) and saturated brine (500 mL) in turn, dried over anhydrous sodium sulfate, and concentrated to obtain intermediate A-2-1c (13.80 g, 43.90 mmol).

[0157] Third step

[0158] Compound intermediate A-2-1c (13.80 g, 43.90 mmol) was dissolved in hydrogen chloride ethyl acetate solution (4.0 M, 100 mL), and the reaction was carried out at room temperature for 2 hours. After the reaction was completed by monitoring by LCMS, it was concentrated, and purified by medium pressure preparation to obtain intermediate A-2-1d (7.56 g, 35.28 mmol, yield 80%).

[0159] Fourth step

[0160] Intermediate A-2-1d (1.0 g, 4.67 mmol) was dissolved in dichloromethane (10 ml). Acetic anhydride (6.7 g, 65.41 mmol) was added and reacted at room temperature for 16 hours, TLC showed the reaction was completed. Cooled to 0 °C and adjusted pH to 2 with 1 M hydrochloric acid. Extracted with ethyl acetate for 3 times (50 mL x 3), combined the organic phase, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, concentrated to get the crude product, and purified by reverse phase preparation to get compound A-2-1 (500 mg, 1.95 mmol, 41.80% yield). MS-ESI calculated value [M+H] + : 257.2; found: 257.0.

[0161] Preparation of compound A-2-2

[0162]

[0163] Intermediate A-2-1d (1.00 g, 4.67 mmol) was dissolved in dry dichloromethane (20 mL), cyclopropylcarbonyl chloride (0.54 g, 5.14 mmol) was added, then triethylamine (1.18 g, 11.68 mmol) was added dropwise. After the addition was completed, it was reacted at room temperature for 1 hour. After the reaction was completed monitored by LCMS, diluted with water (50 mL) and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed with saturated brine in turn, dried over anhydrous sodium sulfate, concentrated, and purified by medium pressure preparation to get compound A-2-2 (0.49 g, 1.74 mmol, 37% yield). MS-ESI calculated value [M+H] + : 283.2; found: 283.3. 1 H NMR (400 MHz, DMSO-d6) δ 8.58 - 8.21 (m, 1H), 5.13 - 4.66 (m, 1H), 4.32 - 4.10 (m, 2H), 3.86 - 3.71 (m, 1H), 3.64 (dd, J = 6.7, 4.2 Hz, 3H), 2.48 - 2.30 (m, 1H), 2.25 - 1.93 (m, 2H), 1.61 - 1.16 (m, 1H), 0.92 - 0.80 (m, 6H), 0.80 - 0.51 (m, 4H).

[0164] Preparation of compound A-2-4

[0165]

[0166] First step

[0167] Compound A-2-1 (0.92 g, 3.59 mmol) and THF (10 mL) were added to a 50 mL dry flask, then sodium hydroxide aqueous solution (2 M, 3 mL) was slowly dropped in, after the addition, the reaction was carried out at room temperature for 2 hours, after the reaction was monitored by LCMS, the reaction solution was adjusted to be acidic (pH = 5) with dilute hydrochloric acid, then extracted with dichloromethane (30 mL x 3). The organic phase was combined, then washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain the crude product of intermediate A-2-4a (0.80 g, 3.31 mmol, yield 92%).

[0168] Second step

[0169] The crude product of intermediate A-2-4a (0.80 g, 3.31 mmol) and deuterated methanol (5 mL) were added to a 50 mL dry flask, then concentrated sulfuric acid (0.36 g, 3.64 mmol) was slowly dropped in, after the addition, the reaction was carried out at room temperature for 2 hours, after the reaction was monitored by LCMS, the reaction solution was adjusted to be neutral (pH = 7) with sodium hydroxide (2.0 M), then extracted with dichloromethane (30 mL x 3). The organic phase was combined, then washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by medium pressure preparation to obtain the product A-2-4 (0.25 g, 0.98 mmol, yield 30%). MS-ESI calculated value [M+H] + : 260.2; found: 260.2. 1 H NMR (400 MHz, DMSO) δ 8.42 (ddd, J = 28.1, 13.8, 6.7 Hz, 1H), 4.97 - 4.64 (m, 1H), 4.32 - 4.13 (m, 1H), 4.08 - 3.93 (m, 1H), 3.83 - 3.62 (m, 1H), 2.41 - 2.29 (m, 1H), 2.16 - 1.91 (m, 2H), 1.83 - 1.56 (m, 3H), 0.96 - 0.77 (m, 6H).

[0170] Preparation of compound A-2-5 of Preparation Example 6

[0171]

[0172] Intermediate A-2-1d (1.00 g, 4.67 mmol), 2-iodopyridine (1.91 g, 9.34 mmol) and N,N-dimethylformamide (30 mL) were added into a 100 mL dry three-necked flask. Then Pd2(dba)3(0.85 g, 0.93 mmol), Xantphos (0.54 g, 0.93 mmol) and cesium carbonate (4.55 g, 14.01 mmol) were added successively. After nitrogen replacement for three times, the reaction was heated to 110 degree for 6 hours. After LCMS monitoring the reaction was completed, water (20 mL) was added for dilution, dichloromethane (50 mL x 3) was used for extraction. The organic phase was combined and washed with water (50 mL) and saturated brine (50 mL) successively, dried over anhydrous sodium sulfate, concentrated and purified by medium pressure preparation to give the product A-2-5 (0.27 g, 0.93 mmol, 20% yield). MS-ESI calculated for [M+H] C17H17IN4, 292.2; found: 292.2. + 1 H NMR (400 MHz, DMSO-d6) δ 8.71 (dd, J = 87.4, 8.3 Hz, 1H), 8.07 (d, J = 4.0 Hz, 1H), 7.63 - 7.52 (m, 1H), 6.72 (ddd, J = 23.9, 12.0, 5.5 Hz, 1H), 6.43 (dd, J = 8.3, 5.8 Hz, 1H), 4.69 (dd, J = 16.5, 8.5 Hz, 1H), 4.29 - 4.20 (m, 1H), 3.95 - 3.72 (m, 2H), 3.64 (d, J = 9.0 Hz, 3H), 2.46 - 2.35 (m, 2H), 2.16 - 2.00 (m, 1H), 0.91 - 0.79 (m, 6H).

[0173] Preparation of compound A-2-6 of Preparation Example 7

[0174]

[0175] First step

[0176] ​A-2-6a (5.00 g, 49.45 mmol), di-tert-butyl dicarbonate (12.95 g, 59.34 mmol) and ethanol (60 mL) were added into a 200 mL dry flask, after stirring well, sodium hydroxide (1.98 g, 49.45 mmol) was dissolved in water (25 mL) and added into the reaction system, and the reaction was carried out at room temperature for 16 hours. After the reaction was completed by monitoring by LCMS, water (60 mL) was added for dilution, and after adjusting to pH = 3 with 1.0 M dilute hydrochloric acid, extraction was carried out with ethyl acetate (150 mL x 3). The organic phase was combined and successively washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain intermediate A-2-6b (8.60 g, 42.74 mmol, yield 86%).

[0177] Second step

[0178] L-valine methyl ester hydrochloride (7.52 g, 44.86 mmol), 4-dimethylaminopyridine (10.45 g, 85.45 mmol) and N,N-dimethylformamide (80 mL) were added into a 200 mL dry flask. After cooling to 0°C, intermediate A-2-6b (8.60 g, 42.74 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (9.83 g, 51.29 mmol) were added, and the reaction was carried out at room temperature for 16 hours. After the reaction was completed by monitoring by LCMS, water (100 mL) was added for dilution, and extraction was carried out with ethyl acetate (200 mL x 3). The organic phase was combined and successively washed with water (500 mL) and saturated brine (500 mL), dried over anhydrous sodium sulfate, and concentrated to obtain intermediate A-2-6c crude product (13.80 g, 43.90 mmol).

[0179] Third step

[0180] The intermediate A-2-6c crude product (13.80 g, 43.90 mmol) was dissolved in hydrogen chloride ethyl acetate solution (4.0 M, 100 mL), and the reaction was carried out at room temperature for 2 hours. After the reaction was completed by monitoring by LCMS, concentration and purification by medium pressure preparative purification were carried out to obtain intermediate A-2-6d (7.56 g, 35.28 mmol, yield 80%).

[0181] Fourth step

[0182] Intermediate A-2-6d (1.00 g, 4.67 mmol) and dichloromethane (15 mL) were added into a 50 mL dry flask, then acetic anhydride (4.76 g, 46.70 mmol) was added slowly, after the addition, the reaction was carried out at room temperature overnight. After monitoring the reaction was completed by LCMS, saturated aqueous sodium bicarbonate solution (50 mL) was added to the reaction solution, stirred for 0.5 h, then extracted with dichloromethane (50 mL x 3). The organic phase was combined, then washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by medium pressure preparation to give the product A-2-6 (0.92 g, 3.59 mmol, 76% yield). MS-ESI calculated value [M+H] + : 257.1; found: 257.2. 1 H NMR (400 MHz, CDCl3) δ 8.31 (d, J = 7.4 Hz, 1H), 4.93 (dd, J = 9.3, 6.5 Hz, 1H), 4.41 (dd, J = 8.1, 5.0 Hz, 1H), 4.08 (t, J = 7.7 Hz, 2H), 3.74 (d, J = 5.0 Hz, 3H), 2.81 - 2.61 (m, 1H), 2.47 - 2.34 (m, 1H), 2.25 - 2.13 (m, 1H), 1.93 (s, 3H), 0.93 (dd, J = 8.5, 6.9 Hz, 6H).

[0183] Preparation of compound A-2-7

[0184]

[0185] First step

[0186] Compound A-2-7a (5.00 g, 49.45 mmol), di-tert-butyl dicarbonate (12.95 g, 59.34 mmol) and ethanol (60 mL) were added into a 200 mL dry flask, after stirring well, sodium hydroxide (1.98 g, 49.45 mmol) was dissolved in water (25 mL) and added to the reaction system, the reaction was carried out at room temperature for 16 hours. After monitoring the reaction was completed by LCMS, diluted with water (60 mL) and adjusted to pH 3 with 1.0 M dilute hydrochloric acid, extracted with ethyl acetate (150 mL x 3). The organic phase was combined, washed with saturated brine in turn, dried over anhydrous sodium sulfate, concentrated to give intermediate A-2-7b (8.60 g, 42.74 mmol, 86% yield).

[0187] Second step

[0188] L-valine methyl ester hydrochloride (7.52 g, 44.86 mmol), 4-dimethylaminopyridine (10.45 g, 85.45 mmol) and N,N-dimethylformamide (80 mL) were added into a 200 mL dry flask, and then intermediate A-2-7b (8.60 g, 42.74 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (9.83 g, 51.29 mmol) were added into the flask at 0 °C, and the mixture was stirred at room temperature for 16 h. After the reaction was completed by LCMS monitoring, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was combined and washed with water (500 mL) and saturated brine (500 mL) successively, dried over anhydrous sodium sulfate, and concentrated to give intermediate A-2-7c (13.80 g, 43.90 mmol).

[0189] Step 3

[0190] The crude intermediate A-2-7c (13.80 g, 43.90 mmol) was dissolved in hydrogen chloride ethyl acetate solution (4.0 M, 100 mL), and the mixture was stirred at room temperature for 2 h. After the reaction was completed by LCMS monitoring, the mixture was concentrated and purified by medium pressure preparative chromatography to give intermediate A-2-7d (7.56 g, 35.28 mmol, 80% yield).

[0191] Step 4

[0192] Intermediate A-2-7d (1.00 g, 4.67 mmol) and dichloromethane (15 mL) were added into a 50 mL dry flask, and then acetic anhydride (4.76 g, 46.70 mmol) was slowly added into the flask. After the addition was completed, the mixture was stirred at room temperature overnight. After the reaction was completed by LCMS monitoring, saturated sodium bicarbonate aqueous solution (50 mL) was added into the mixture, and the mixture was stirred for 0.5 h. The mixture was extracted with dichloromethane (50 mL x 3). The organic phase was combined and washed with water (500 mL) and saturated brine (500 mL) successively, dried over anhydrous sodium sulfate, concentrated, and purified by medium pressure preparative chromatography to give product A-2-7 (0.92 g, 3.59 mmol, 76% yield). MS-ESI calculated for [M+H] C17H23NO5: 257.1; found: 257.2. + 1 ​H NMR (400 MHz, CDC13) δ 8.28 (d, J = 7.9 Hz, 1H), 4.91 (dd, J = 9.5, 6.6 Hz, 1H), 4.48 (dd, J = 8.4, 4.9 Hz, 1H), 4.13 - 4.00 (m, 2H), 3.73 (s, 3H), 2.73 - 2.58 (m, 1H), 2.52 - 2.35 (m, 1H), 2.30 - 2.15 (m, 1H), 1.95 (s, 3H), 0.96 (dd, J = 16.1, 6.9 Hz, 6H).

[0193] Preparation of compound A-2-8 of Preparation Example 9

[0194]

[0195] Intermediate A-2-6d (1.4 g, 6.5 mmol) was dissolved in dichloromethane (10 mL), N,N-diisopropylethylamine (3.38 g, 26.2 mmol) was added, nitrogen was replaced for three times, and deuterated acetyl chloride (0.58 g, 7.19 mmol) was added dropwise after the reaction was reduced to 0°C. After the reaction liquid was stirred at 20°C for 1 hour, saturated sodium bicarbonate solution (20 mL) was added, dichloromethane was extracted three times (50 mL x 3). The organic phase was combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane / ethyl acetate = 1 / 1) to obtain the product A-2-8 (1.31 g, 5.05 mmol, yield 77.3%). MS-ESI calculated value [M+H] + : 260.2; found: 260. 1 H NMR (400 MHz, CDC13) δ 8.31 (d, J = 7.2 Hz, 1H), 4.93 (dd, J = 9.2, 6.5 Hz, 1H), 4.41 (dd, J = 8.0, 5.0 Hz, 1H), 4.08 (t, J = 7.7 Hz, 2H), 3.73 (s, 3H), 2.71 (ddd, J = 19.2, 13.4, 7.7 Hz, 1H), 2.41 (ddd, J = 16.3, 12.0, 7.8 Hz, 1H), 2.20 (td, J = 13.6, 6.8 Hz, 1H), 0.93 (dd, J = 8.3, 7.1 Hz, 6H).

[0196] Preparation of compound A-2-9 of Preparation Example 10

[0197]

[0198] Intermediate A-2-7d (1.5 g, 7.0 mmol) was dissolved in dichloromethane (20 mL), N, N-diisopropyl ethylamine (4.52 g, 35 mmol) was added, and the reaction was bubbled with nitrogen three times. After the reaction was reduced to 0 °C, deuterated acetyl chloride (630 mg, 7.7 mmol) was added dropwise. After the reaction was stirred at room temperature for 1 h, saturated aqueous sodium bicarbonate solution (20 mL) was added, and extracted with dichloromethane (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2:1) to give the product A-2-9 (1.1 g, 4.24 mmol, 60.6% yield). MS-ESI [M+H] calculated: 260.2; found: 260.1. + 1 H NMR (400 MHz, CDCl3) δ 8.28 (d, J = 7.4 Hz, 1H), 4.91 (dd, J = 9.4, 6.6 Hz, 1H), 4.47 (dd, J = 8.4, 4.9 Hz, 1H), 4.12-4.03 (m, 2H), 3.72 (s, 3H), 2.70-2.60 (m, 1H), 2.47-2.38 (m, 1H), 2.28-2.15 (m, 1H), 0.95 (dd, J = 16.2, 6.9 Hz, 6H).

[0199] Preparation of compound A-2-10

[0200]

[0201] Compound A-2-10 is intermediate A-2-1d, the preparation of which is described in Preparation Example 3.

[0202] Preparation of compound A-2-11

[0203]

[0204] Compound A-2-11 is intermediate A-2-6d, the preparation of which is described in Preparation Example 7.

[0205] Preparation of compound A-2-12

[0206]

[0207] Compound A-2-12 is intermediate A-2-7d, the preparation of which is described in Preparation Example 8.

[0208] Preparation of compound A-2-15

[0209]

[0210] ​First step

[0211] Compound A-2-15a (480 mg, 3.69 mmol) and L-valine methyl ester hydrochloride (680 mg, 4.06 mmol, 1.1 eq) were dissolved in anhydrous N,N-dimethylformamide (10 mL), 4-dimethylaminopyridine (900 mg, 7.38 mmol) and l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (780 mg, 4.06 mmol) were added successively at 0 °C, after stirring for 1 h at 0 °C, it was raised to room temperature, and the reaction was allowed to proceed overnight. A small amount of water was added to quench, and after removing N,N-dimethylformamide by concentration, it was dissolved with water and acetonitrile, and compound A-2-15 (347 mg, 1.43 mmol, 38.6% yield) was obtained by medium pressure preparation. MS-ESI calculated value [M+H] + : 244.1; found 244.1.

[0212] Preparation of compound A-2-16 of Preparation Example 15

[0213]

[0214] First step

[0215] Compound A-2-16a (440 mg, 3.07 mmol) and L-valine methyl ester hydrochloride (567 mg, 3.38 mmol, 1.1 eq) were dissolved in anhydrous N,N-dimethylformamide (10 mL), 4-dimethylaminopyridine (750 mg, 6.14 mmol) and l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (650 mg, 3.38 mmol) were added successively at 0 °C, after stirring for 1 h at 0 °C, it was raised to room temperature, and the reaction was allowed to proceed overnight. A small amount of water was added to quench, and after removing N,N-dimethylformamide by concentration, it was dissolved with water and acetonitrile, and compound A-2-16 (465 mg, 1.81 mmol, 59.0% yield) was obtained by medium pressure preparation. MS-ESI calculated value [M+H] + : 257.2; found 257.0.

[0216] Preparation of compound A-2-17 of Preparation Example 16

[0217]

[0218] First step

[0219] Compound A-2-17a (10.0 g, 41.9 mmol) was dissolved in N,N-dimethylformamide (80 mL), potassium carbonate (14.3 g, 103.47 mmol) was added thereto, and then (2-aminoethyl) benzylcarbamate (12.2 g, 62.9 mmol) was added at 0°C. The reaction solution was reacted at 100°C for 6 hours. After the reaction was completed, water (50 mL) was added to the reaction solution, and then extracted with ethyl acetate (50 mL x 3), and the organic phase was combined, washed with brine (100 mL x 3), dried over anhydrous sodium sulfate, and concentrated. The obtained residue was separated and purified with a silica gel column (petroleum ether / ethyl acetate = 3 / 1) to obtain compound A-2-17b (5.8 g, yield: 39%). MS-ESI calculated value [M+H]: 353.17; found: 353.16. + : 353.17; found: 353.16.

[0220] Second step

[0221] Compound A-2-17b (5.8 g, 16.5 mmol) was dissolved in methanol (40 mL), and palladium carbon (580 mg) was added thereto. The reaction solution was reacted at 25°C for 12 hours under a hydrogen atmosphere. After the reaction was completed, the palladium carbon was removed by filtration, di-tert-butyl dicarbonate (5.4 g, 24.8 mmol) was added to the filtrate, and reacted at room temperature for 2 hours. After the reaction was completed, it was concentrated, and the obtained residue was separated and purified with a silica gel column (dichloromethane / methanol = 20 / 1) to obtain compound A-2-17c (3.0 g, yield: 62%). MS-ESI calculated value [M+H]: 273.14; found: 272.85. + : 273.14; found: 272.85.

[0222] Third step

[0223] Compound A-2-17c (3.0 g, 11 mmol) was dissolved in methanol (30 mL), and lithium hydroxide (0.4 g, 16.5 mmol) was added thereto. The reaction solution was stirred at room temperature for 2 hours. Then, the reaction solution was lyophilized and recrystallized with methanol to obtain compound A-2-17d (2.6 g, yield: 96.6%). MS-ESI calculated value [M+H]: 245.11; found: 245.11. + : 245.11; found: 245.11.

[0224] Fourth step:

[0225] Compound A-2-17d (2.6 g, 10.6 mmol) was dissolved in N,N-dimethylformamide (30 mL), and L-valine methyl ester hydrochloride (3.54 g, 21.2 mmol), 1-hydroxybenzotriazole (2.86 g, 21.2 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.05 g, 21.2 mmol) and N,N-diisopropylethylamine (4.12 g, 31.8 mmol) were added thereto. The reaction solution was allowed to react at room temperature for 4 hours. After completion of the reaction, water (50 mL) was added to the reaction solution, which was then extracted with ethyl acetate (50 mL x 3), and the organic phase was combined, washed with brine (100 mL x 3), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was separated and purified by silica gel column chromatography (dichloromethane / methanol = 15 / 1), and then by preparative HPLC to obtain two isomers of the target product, compound A-2-17e1 (300 mg) and A-2-17e2 (200 mg). MS-ESI calculated value [M+H] + : 358.20; found: 358.02.

[0226] Fifth step:

[0227] Compound A-2-17e1 (300 mg, 0.84 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (2 mL) was added thereto, respectively. The reaction solution was stirred at room temperature for 2 hours. After completion of the reaction, the solvent was removed by rotary evaporation, and freeze-drying to obtain compound A-2-17A (155 mg). MS-ESI calculated value [M+H] + : 258.14; found: 258.05. 1 H NMR (400 MHz, DMSO-d6) δ 9.34 (s, 2H), 8.82-8.80 (m, 1H), 8.66-8.61 (m, 1H), 4.81-4.75 (m, 1H), 4.30-4.27 (m, 1H), 3.68-3.66 (m, 3H), 3.42-3.32 (m, 4H), 2.13-2.07 (m, 1H), 0.91-0.89 (m, 6H).

[0228] Compound A-2-17e2 (200 mg, 0.56 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (2 mL) was added thereto, respectively. The reaction solution was stirred at room temperature for 2 hours. After completion of the reaction, the solvent was removed by rotary evaporation, and compound A-2-17B (73 mg) was obtained by preparative HPLC. MS-ESI calculated value [M+H] + : 258.14; found: 258.00. 1H NMR (400 MHz, DMSO-d6) δ 9.34 (s, 1H), 8.81 (d, J = 8.4 Hz, 1H), 8.63 (d, J = 18.0 Hz, 1H), 4.781 (d, J = 26.4 Hz, 1H), 4.33-4.27 (m, 1H), 3.68-3.67 (m, 3H), 3.42-3.32 (m, 4H), 2.13-2.07 (m, 1H), 0.91-0.89 (m, 6H).

[0229] Preparation of compound A-2-20

[0230]

[0231] First step

[0232] Compound A-2-7a (10.00 g, 98.91 mmol) was dissolved in ethanol (100 mL), then di-tert-butyl dicarbonate (43.17 g, 197.82 mmol) was added, 4-dimethylaminopyridine (1.74 g, 14.27 mmol) was added slowly in batches at room temperature, and sodium hydroxide (3.96 g, 98.91 mmol) was dissolved in water (100 mL) and added to the above reaction system, and stirred at room temperature. After 4 hours, TLC monitoring showed that the reaction was complete. The above reaction system was diluted with water (100 mL) and adjusted to pH 3 with 1M hydrochloric acid. Extracted with ethyl acetate (200 mL x 3). The organic phase was combined and washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, concentrated to give the crude intermediate A-2-7b (22.10 g). MS-ESI [M-H] Theoretical value: 200.1; Found: 200.1. -

[0233] Second step

[0234] ​In a single-necked flask, intermediate A-2-7b (10.00 g, 49.70 mmol) and L-valine methyl ester hydrochloride (10.00 g, 59.64 mmol) were added and dissolved in dichloromethane (150 mL). Then, O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (22.68 g, 59.64 mmol) and triethylamine (10.06 g, 99.40 mmol) were added successively. After stirring at room temperature for 16 h, TLC showed that the reaction was complete. The reaction was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was washed successively with water (500 mL x 3) and saturated brine (500 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 3 / 2) to give intermediate A-2-7c (13.84 g, 44.02 mmol, 88.6% yield).

[0235] Third step

[0236] In a single-necked flask, intermediate A-2-7c (3.00 g, 9.54 mmol) was added, followed by the addition of ethyl acetate hydrochloride solution (3.0 M, 5 mL). After stirring at room temperature for 4 h, the reaction was complete as shown by TCL, and was concentrated to give intermediate A-2-20a (2.02 g, 8.07 mmol, 84.4% yield). MS-ESI [M+H] calc'd for C9H13N3O2: 215.1; found: 215.3. +

[0237] Fourth step

[0238] In a dry three-necked flask, cyclopropyl carboxylic acid (0.12 g, 1.44 mmol) and N,N,N,N-tetramethyluronium hexafluorophosphate (0.67 g, 2.40 mmol), N-methylimidazole (0.20 g, 2.40 mmol), and triethylamine (0.20 g, 1.98 mmol) were added. Dichloromethane (5 mL) was added, and after stirring for 0.5 h, intermediate A-2-20a (0.30 g, 1.20 mmol) was added and reacted at room temperature for 2 h. After the reaction was completed, the solvent was evaporated, water (10 mL) was added, and extraction was performed with ethyl acetate (10 mL x 3). The organic phase was combined and washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give product A-2-20 (135.3 mg, 0.48 mmol, 40.1% yield). MS-ESI [M+H] calc'd for C14H19N5O2: 283.2; found: 283.3. + 1 ​​H NMR (400 MHz, Chloroform-d) δ 8.25 (d, J = 8.5 Hz, 1H), 4.91 (dd, J = 9.5, 6.5 Hz, 1H), 4.47 (dd, J = 8.2, 4.8 Hz, 1H), 4.20 (q, J = 6.5, 4.5 Hz, 2H), 3.72 (s, 3H), 2.67 (d, J = 9.6 Hz, 1H), 2.59 - 2.37 (m, 1H), 2.30 - 2.08 (m, 1H), 1.44 (tq, J = 11.2, 6.9, 5.7 Hz, 1H), 1.03 (dt, J = 11.7, 5.2 Hz, 2H), 0.93 (dd, J = 20.5, 6.9 Hz, 6H), 0.87 - 0.78 (m, 2H).

[0239] Preparation of compound A-2-21

[0240]

[0241] First step

[0242] In a dry three-necked flask, compound A-2-6a (5.20 g, 51.43 mmol) was added to sodium hydroxide aqueous solution (1 M, 50 mL) at 0 °C, and after dissolution, ethanol (50 mL) was added. Di-tert-butyl dicarbonate (13.47 g, 61.72 mmol) was added dropwise at this temperature, and the reaction was stirred at room temperature overnight. After the reaction was completed, the pH was adjusted to 4-5 with saturated citric acid solution, extracted with dichloromethane (100 mL x 3), the combined organic phase was washed with saturated sodium chloride solution (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain intermediate A-2-6b (9.59 g, 47.66 mmol, yield 92.7%). MS-ESI [M-H]- - : 200.1 ; Found: 200.1. 1 H NMR (400 MHz, Chloroform-d) δ 9.19 (s, 1H), 4.77 (s, 1H), 3.94 (q, J = 7.7 Hz, 2H), 2.48 (s, 2H), 1.46 (s, 9H).

[0243] Second step

[0244] In a dry three-necked flask, N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (22.68 g, 59.64 mmol), triethylamine (10.00 g, 99.40 mmol) were dissolved in dichloromethane (100 mL) at 0 °C. Intermediate A-2-6b (9.59 g, 47.66 mmol) was added, and after stirring for half an hour, L-valine methyl ester hydrochloride (7.82 g, 59.64 mmol) was added. The reaction was carried out at room temperature for 16 hours. After the reaction was completed, water (100 mL) was added, and extraction was carried out with ethyl acetate (100 mL x 3). The organic phase was combined and washed with saturated brine (100 mL x 2) in turn, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane / methanol = 15 / 1) to obtain intermediate A-2-6c (11.50 g, 36.58 mmol, 76.8% yield). MS-ESI [M-H] - : 313.2; Found: 313.0. 1 H NMR (400 MHz, DMSO-d6) δ 8.17 (d, J = 8.3 Hz, 1H), 4.72 - 4.50 (m, 1H), 4.22 (dd, J = 8.2, 6.1 Hz, 1H), 3.85 - 3.66 (m, 2H), 3.64 (s, 3H), 2.36 (s, 1H), 2.13 - 1.82 (m, 2H), 1.42 - 1.27 (m, 9H), 0.88 (d, J = 6.8 Hz, 6H).

[0245] Third step

[0246] In a dry three-necked flask, intermediate A-2-6c (2.50 g, 7.95 mmol) was dissolved in 5 mL of dichloromethane solution at 0 °C, and hydrogen chloride in dioxane (4 M, 20 mL) was added dropwise. After the addition was completed, it was moved to room temperature and stirring was continued. After the reaction was completed, it was concentrated to obtain intermediate A-2-21a (2.00 g, 7.95 mmol, 100%). MS-ESI [M+H] + : 215.1; Found: 215.3.

[0247] Fourth step

[0248] In a dry three-necked flask, cyclopropylcarboxylic acid (0.15 g, 1.20 mmol) and 3-(ethylcarbamoylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.23 g, 1.20 mmol), 1-hydroxybenzotriazole (0.16 g, 1.20 mmol) and triethylamine (0.20 g, 1.60 mmol) were added. Dichloromethane (5 mL) was added, the reaction was stirred for 0.5 h, and intermediate A-2-21a (0.25 g, 1.00 mmol) was added. The reaction was allowed to proceed at room temperature for 2 h. After the reaction was completed, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phase was combined, washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give the product A-2-21 (99.1 mg, 0.35 mmol, yield: 35.2%). MS-ESI [M+H] calc’d for C19H29N4O4: 383.2; found: 383.3. + 1 H NMR (400 MHz, Chloroform-d) δ 8.42 (d, J = 8.2 Hz, 1H), 4.93 (dd, J = 9.4, 6.4 Hz, 1H), 4.40 (dd, J = 8.2, 5.0 Hz, 1H), 4.20 (ddd, J = 12.0, 8.9, 4.8 Hz, 2H), 3.63 (s, 3H), 2.74 (ddd, J = 12.5, 9.2, 6.2 Hz, 1H), 2.44 (dtd, J = 11.9, 9.2, 5.5 Hz, 1H), 2.18 (td, J = 7.2, 5.3 Hz, 1H), 1.42 (td, J = 8.0, 4.1 Hz, 1H), 1.01 (q, J = 3.8 Hz, 2H), 0.90 (dd, J = 16.3, 6.9 Hz, 6H), 0.82 (dtd, J = 9.3, 5.9, 2.7 Hz, 2H).

[0249] Preparation of compound A-2-22

[0250]

[0251] ​In a single-necked flask, benzoic acid (146.5 mg, 1.20 mmol), l-ethyl-(3- dimethylaminopropyl)carbodiimide hydrochloride (299.0 mg, 1.56 mmol), 1- hydroxybenzotriazole (210.8 mg, 1.56 mmol) were added and dissolved in dichloromethane (5 mL) and stirred at 0 °C for 0.5 h. Then, intermediate A-2-20a (300.0 mg, 1.20 mmol) and diisopropylethylamine (465.3 mg, 3.60 mmol) were added sequentially and stirred at room temperature overnight. The reaction progress was monitored by TLC. After TLC showed the reaction was complete, the reaction was quenched by adding water (10 mL), extracted and separated. Then, the aqueous phase was further extracted with dichloromethane (10 mL x 2), the organic phase was collected, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (EA: PE = 10% ~ 60%) to give product A-2-22 (227.0 mg, 0.71 mmol, 59.59% yield). MS-ESI [M+H] Calc. for C21H25N4O4: 419.2; Found: 419.3. + 1 H NMR (400 MHz, Chloroform-d) δ 7.61-7.57 (m, 2H), 7.46-7.41 (m, 1H), 7.37 (dd, J = 8.1, 6.5 Hz, 2H), 5.09-4.98 (m, 1H), 4.47 (dd, J = 8.6, 4.9 Hz, 1H), 4.30 (td, J = 8.9, 5.6 Hz, 1H), 4.06 (q, J = 8.1 Hz, 1H), 3.64 (s, 3H), 2.69 (s, 1H), 2.43 (s, 1H), 2.23-2.09 (m, 1H), 0.89 (dd, J = 21.6, 6.9 Hz, 6H).

[0252] Preparation of compound A-2-23

[0253]

[0254] ​In a dry three-necked flask, add benzoic acid (0.19 g, 1.54 mmol) and 3- (ethylcarbamoylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.29 g, 1.54 mmol), 1-hydroxybenzotriazole (0.21 g, 1.54 mmol) and triethylamine (0.26 g, 2.56 mmol). Add dichloromethane (5 mL), after stirring for 0.5 h, add intermediate A-2-21a (0.32 g, 1.28 mmol) and react at room temperature for 2 h. After the reaction is completed, add water (10 mL) and extract with ethyl acetate (10 mL x 3). Wash the combined organic phase with saturated brine (10 mL x 2), dry over anhydrous sodium sulfate, filter and concentrate. Purify by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain the product A-2-23 (69.0 mg, 0.22 mmol, yield: 17.0%). MS-ESI [M+H] calculated for C25H31N5O4: 479.2; found: 479.3. + 1 H NMR (400 MHz, Chloroform-d) δ 8.09-7.85 (m, 1H), 7.65-7.60 (m, 2H), 7.50 (t, J = 7.2 Hz, 1H), 7.43 (t, J = 7.4 Hz, 2H), 5.12 (t, J = 8.1 Hz, 1H), 4.48 (dd, J = 8.4, 5.0 Hz, 1H), 4.36 (td, J = 8.9, 5.6 Hz, 1H), 4.14 (q, J = 8.2 Hz, 1H), 3.74 (s, 3H), 2.84 (s, 1H), 2.44 (s, 1H), 2.29-2.09 (m, 1H), 0.92 (dd, J = 12.3, 6.9 Hz, 6H).

[0255] Preparation of compound A-2-24

[0256]

[0257] ​In a single-necked flask, 4-trifluoromethylbenzoic acid (228.1 mg, 1.20 mmol), l-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.30 g, 1.56 mmol), 1-hydroxybenzotriazole (0.21 g, 1.56 mmol) were added and dissolved in dichloromethane (5 mL) and stirred at 0 °C for 0.5 h. Then, intermediate A-2-20a (300.0 mg, 1.20 mmol) and diisopropylethylamine (0.47 g, 3.60 mmol) were added sequentially and stirred at room temperature overnight, monitoring the reaction progress by TLC. After TLC showed the reaction was complete, the reaction was quenched by adding water (10 mL), extracted and separated, then the aqueous phase was further extracted with dichloromethane (10 mL x 2), the organic phase was collected, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (EA:PE = 10%~60%) to give the product A-2-24 (218.0 mg, 0.56 mmol, 47.15% yield). MS-ESI [M+H] Calc’d for C26H28F3N4O4: 387.1; Found: 387.4. + 1 H NMR (400 MHz, Chloroform-d) δ 7.86 (d, J = 7.2 Hz, 1H), 7.77 (d, J = 8.2 Hz, 2H), 7.70 (d, J = 8.2 Hz, 2H), 5.16 - 5.07 (m, 1H), 4.54 (dd, J = 8.6, 4.8 Hz, 1H), 4.36 (td, J = 8.9, 5.7 Hz, 1H), 4.12 (q, J = 8.2 Hz, 1H), 3.70 (s, 3H), 2.79 (s, 1H), 2.61 - 2.45 (m, 1H), 2.23 (q, J = 6.6 Hz, 1H), 0.96 (dd, J = 23.5, 6.9 Hz, 6H).

[0258] Preparation of compound A-2-25 of Preparation Example 22

[0259]

[0260] ​In a dry three-necked flask, 4-trifluoromethylbenzoic acid (0.18 g, 0.96 mmol) and 3-(ethylcarbamoylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.18 g, 0.96 mmol), 1-hydroxybenzotriazole (0.13 g, 0.96 mmol) and triethylamine (0.16 g, 1.60 mmol) were added. Dichloromethane (5 mL) was added and the reaction was stirred for 0.5 h. Intermediate A-2-21a (0.20 g, 0.80 mmol) was added. The reaction was stirred at room temperature for 2 h, water (10 mL) was added and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phase was combined, washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give the product A-2-25 (130.0 mg, 0.34 mmol, yield: 42.2%). MS-ESI [M+H] calculated for C25H30F3N4O4: 487.2; found: 487.4. + 1 H NMR (400 MHz, Chloroform-d) δ 7.88 (d, J = 8.4 Hz, 1H), 7.73 (q, J = 8.3 Hz, 4H), 5.13 (dd, J = 9.4, 6.4 Hz, 1H), 4.49 (dd, J = 8.4, 4.9 Hz, 1H), 4.37 (td, J = 9.0, 5.6 Hz, 1H), 4.22 - 4.03 (m, 1H), 3.75 (s, 3H), 2.85 (d, J = 16.4 Hz, 1H), 2.48 (d, J = 10.0 Hz, 1H), 2.21 (q, J = 6.6 Hz, 1H), 0.92 (dd, J = 13.2, 6.9 Hz, 6H).

[0261] Preparation of compound A-2-26

[0262]

[0263] ​In a single neck flask, p-fluorobenzoic acid (168.1 mg, 1.20 mmol), l-ethyl-(3- dimethylaminopropyl)carbodiimide hydrochloride (300 mg, 1.56 mmol), 1- hydroxybenzotriazole (0.21 g, 1.56 mmol) were added and dissolved in dichloromethane (5 mL) and stirred at 0 °C for 0.5 h. Then, intermediate A-2-20a (300.0 mg, 1.20 mmol) and diisopropylethylamine (0.47 g, 3.60 mmol) were added sequentially and stirred at room temperature overnight, monitoring the reaction progress by TLC. After TLC showed the reaction was complete, the reaction was quenched by the addition of water (10 mL), extracted and partitioned, then the aqueous phase was further extracted with dichloromethane (10 mL x 2), the organic phase was collected, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (= 10% to 60%) to give the product A-2-26 (270.0 mg, 0.88 mmol, 59.34% yield). MS-ESI [M+H] Calc’d: 337.2; Found: 337.4. + 1 H NMR (400 MHz, Chloroform-d) δ 7.85 (s, 1H), 7.62 (dd, J = 8.6, 5.3 Hz, 2H), 7.05 (t, J = 8.5 Hz, 2H), 5.08 - 4.99 (m, 1H), 4.47 (dd, J = 8.6, 4.9 Hz, 1H), 4.30 (td, J = 8.8, 5.5 Hz, 1H), 4.14 - 4.02 (m, 1H), 3.64 (s, 3H), 2.70 (s, 1H), 2.44 (s, 1H), 2.15 (dq, J = 13.9, 6.7 Hz, 1H), 0.89 (dd, J = 21.8, 6.9 Hz, 6H).

[0264] Preparation of compound A-2-27

[0265]

[0266] ​In a dry three-necked flask, 4-fluorobenzoic acid (0.13 g, 0.96 mmol) and 3- (ethylcarbamoylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.18 g, 0.96 mmol), 1-hydroxybenzotriazole (0.13 g, 0.96 mmol) and triethylamine (0.16 g, 1.60 mmol) were charged. Dichloromethane (5 mL) was added, the reaction was stirred for 0.5 h, and intermediate A-2-21a (0.20 g, 0.80 mmol) was added. The reaction was allowed to proceed at room temperature for 2 h, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give the product A-2-27 (99.4 mg, 0.28 mmol, yield: 35.3%). MS-ESI [M+H] calc’d for C25H30FN4O4: 477.2; found: 477.4. + 1 H NMR (400 MHz, Chloroform-d) δ 7.93 (s, 1H), 7.75-7.62 (m, 2H), 7.12 (t, J = 8.6 Hz, 2H), 5.11 (s, 1H), 4.47 (dd, J = 8.4, 5.0 Hz, 1H), 4.42-4.29 (m, 1H), 4.15 (d, J = 7.9 Hz, 1H), 3.74 (s, 3H), 2.84 (s, 1H), 2.44 (s, 1H), 2.20 (pd, J = 7.0, 5.1 Hz, 1H), 0.91 (dd, J = 11.7, 6.9 Hz, 6H).

[0267] Preparation of compound A-2-28

[0268]

[0269] ​In a single-necked flask, p-chlorobenzoic acid (187.9 mg, 1.20 mmol), l-ethyl-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.30 g, 1.56 mmol), 1-hydroxybenzotriazole (0.21 g, 1.56 mmol) were added and dissolved in dichloromethane (5 mL) and stirred at 0 °C for 0.5 h. Then, intermediate A-2-20a (300.0 mg, 1.20 mmol) and diisopropylethylamine (0.47 g, 3.60 mmol) were added sequentially and stirred at room temperature overnight, monitoring the reaction progress by TLC. After TLC showed the reaction was complete, the reaction was quenched by adding water (10 mL), extracted and separated, then the aqueous phase was further extracted with dichloromethane (10 mL x 2), the organic phase was collected, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (EA:PE = 10%~60%) to give the product A-2-28 (218.0 mg, 0.56 mmol, 47.15% yield). MS-ESI [M+H] calc’d for C21H25ClN4O4: 433.1; found: 433.4. + 1 H NMR (400 MHz, Chloroform-d) δ 7.83 (s, 1H), 7.58 (d, J = 8.3 Hz, 2H), 7.39 (d, J = 8.2 Hz, 2H), 5.11 - 5.02 (m, 1H), 4.50 (dd, J = 8.6, 4.9 Hz, 1H), 4.33 (td, J = 8.9, 5.6 Hz, 1H), 4.11 (q, J = 7.5, 6.5 Hz, 1H), 3.68 (s, 3H), 2.74 (s, 1H), 2.48 (s, 1H), 2.19 (h, J = 6.8 Hz, 1H), 0.93 (dd, J = 22.1, 6.9 Hz, 6H).

[0270] Preparation of compound A-2-29

[0271]

[0272] ​In a dry three-necked flask, 4-chlorobenzoic acid (0.30 g, 1.92 mmol) and 3-(ethylcarbamoylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.37 g, 1.92 mmol), 1-hydroxybenzotriazole (0.26 g, 1.92 mmol) and triethylamine (0.32 g, 3.20 mmol) were added. Dichloromethane (5 mL) was added, the reaction was stirred for 0.5 h, and intermediate A-2-21a (0.40 g, 1.60 mmol) was added. The reaction was allowed to proceed at room temperature for 2 h, water (10 mL) was added, and extraction was performed with ethyl acetate (10 mL x 3 L). The organic phase was combined, washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain the product A-2-29 (64.6 mg, 0.18 mmol, yield: 11.5%). MS-ESI [M+H] calculated for C21H25ClN4O4, 423.1; found, 423.3. + 1 H NMR (400 MHz, Chloroform-d) δ 7.91 (d, J = 8.3 Hz, 1H), 7.77 - 7.51 (m, 2H), 7.41 (d, J = 8.3 Hz, 2H), 5.10 (t, J = 8.0 Hz, 1H), 4.47 (dd, J = 8.3, 5.0 Hz, 1H), 4.35 (q, J = 8.9, 8.4 Hz, 1H), 4.15 (t, J = 8.1 Hz, 1H), 3.74 (s, 3H), 2.84 (s, 1H), 2.44 (s, 1H), 2.29 - 2.05 (m, 1H), 0.91 (dd, J = 12.0, 6.8 Hz, 6H).

[0273] Preparation of compound A-2-30

[0274]

[0275] ​In a single-neck flask, furan-2-carboxylic acid (165.9 mg, 1.48 mmol), l-ethyl-(3- dimethylaminopropyl)carbodiimide hydrochloride (0.37 g, 1.92 mmol), 1-hydroxybenzotriazole (0.26 g, 1.92 mmol) were added and dissolved in dichloromethane (5 mL), stirred at 0 °C for 0.5 h. Then, compound intermediate A-2-20a (370.0 mg, 1.48 mmol) and diisopropylethylamine (0.57 g, 4.44 mmol) were added sequentially, stirred at room temperature overnight, and the reaction progress was monitored by TLC. After TLC showed that the reaction was complete, the reaction was quenched by adding water (10 mL), extracted and separated, then the aqueous phase was further extracted with dichloromethane (10 mL x 2), the organic phase was collected, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography (EA: PE = 10% ~ 60%) to obtain compound A-2-30 (270.0 mg, 0.88 mmol, yield 59.34%). MS-ESI [M+H] Theoretical value: 309.1; Found: 309.3. + 1 H NMR (400 MHz, Chloroform-d) δ 8.09 (s, 1H), 7.46 (s, 1H), 7.08 (d, J = 3.5 Hz, 1H), 6.45 (dd, J = 3.5, 1.7 Hz, 1H), 5.03 (t, J = 8.4 Hz, 1H), 4.44 (dd, J = 14.4, 7.2 Hz, 3H), 3.64 (s, 3H), 2.70 (s, 1H), 2.49 (s, 1H), 2.24 - 2.10 (m, 1H), 1.09 - 0.84 (m, 6H).

[0276] Preparation of compound A-2-31

[0277]

[0278] ​In a dry three-necked flask, 2-furancarboxylic acid (0.13 g, 1.20 mmol) and 3- (ethylcarbamoylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.23 g, 1.20 mmol), 1-hydroxybenzotriazole (0.16 g, 1.20 mmol) and triethylamine (0.20 g, 1.60 mmol) were added. Dichloromethane (5 mL) was added, the reaction was stirred for 0.5 h, and intermediate A-2-21a (0.25 g, 1.00 mmol) was added. After reaction at room temperature for 2 h, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phase was combined, washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give the product A-2-31 (182.2 mg, 0.57 mmol, yield: 57.2%). MS-ESI [M+H] calculated for C19H23N4O5: 393.2; found: 393.3. + 1 H NMR (400 MHz, Chloroform-d) δ 8.25 (s, 1H), 7.53 (s, 1H), 7.14 (d, J = 3.5 Hz, 1H), 6.51 (dd, J = 3.6, 1.8 Hz, 1H), 5.21 - 4.93 (m, 1H), 4.47 (d, J = 13.1 Hz, 3H), 3.73 (s, 3H), 2.84 (s, 1H), 2.50 (s, 1H), 2.19 (td, J = 6.9, 5.1 Hz, 1H), 0.92 (dd, J = 8.1, 6.8 Hz, 6H).

[0279] Preparation of compound A-2-32

[0280]

[0281] ​In a single neck flask, thiophene-2-carboxylic acid (306.0 mg, 2.39 mmol) was dissolved in dichloromethane (10 mL). Then, compound intermediate A-2-20a (500.0 mg, 1.99 mmol), 2-(7-azabenzotriazolyl)-tetramethyluronium hexafluorophosphate (500.0 mg, 2.59 mmol) and triethylamine (402 mg, 3.98 mmol) were added sequentially and stirred at room temperature overnight. The reaction was monitored by TLC. After TLC showed the reaction was complete, the reaction was quenched by the addition of water (10 mL), extracted and partitioned. The aqueous phase was further extracted with dichloromethane (10 mL x 2), the organic phase was collected, washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate and concentrated to give the crude product. The crude product was purified by silica gel column chromatography (EA: PE = 10% to 60%) to give product A-2-32 (302.2 mg, 0.95 mmol, 46.7% yield). MS-ESI [M+H] calc’d for C15H15F2N3O2: 325.1; found: 325.3. + 1 H NMR (400 MHz, Chloroform-d) δ 8.09 (s, 1H), 7.52 (dd, J = 7.5, 4.2 Hz, 2H), 7.16 - 6.98 (m, 1H), 5.08 (dd, J = 9.4, 6.5 Hz, 1H), 4.54 - 4.24 (m, 3H), 3.66 (s, 3H), 2.78 (s, 1H), 2.57 (d, J = 27.1 Hz, 1H), 2.17 (dp, J = 13.5, 6.8 Hz, 1H), 0.92 (dd, J = 17.5, 6.6 Hz, 6H).

[0282] Preparation of compound A-2-33

[0283]

[0284] ​In a dry three-necked flask, add 2-thiophene carboxylic acid (0.15 g, 1.20 mmol) and 3-(ethylcarbamoylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.23 g, 1.20 mmol), 1-hydroxybenzotriazole (0.16 g, 1.20 mmol) and triethylamine (0.20 g, 1.60 mmol). Add dichloromethane (5 mL), stir the reaction for 0.5 h, and then add intermediate A-2-21a (0.25 g, 1.00 mmol). After reaction at room temperature for 2 h, add water (10 mL) and extract with ethyl acetate (10 mL x 3). Combine the organic phases, wash with saturated brine (10 mL x 2), dry over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain the product A-2-33 (77.2 mg, 0.24 mmol, yield: 23.9%). MS-ESI [M+H] calculated for C21H25N4O3S, 425.1; found, 425.2. + : 325.1; found, 325.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.16 (s, 1H), 7.75 - 7.37 (m, 2H), 7.12 (dd, J = 5.0, 3.8 Hz, 1H), 5.13 (dd, J = 9.4, 6.3 Hz, 1H), 4.41 (dtd, J = 17.3, 8.4, 5.9 Hz, 3H), 3.73 (s, 3H), 2.86 (s, 1H), 2.51 (s, 1H), 2.20 (pd, J = 6.9, 5.1 Hz, 1H), 0.92 (t, J = 6.7 Hz, 6H).

[0285] Preparation of compound A-2-34

[0286]

[0287] First step

[0288] In a dry single-necked flask, add compound A-2-34a (3 g, 13.81 mmol) and dichloromethane (50 ml), and stir in an ice bath for 10 min. Then add cyclopentylmethanol (2.07 g, 20.71 mmol), 4-dimethylaminopyridine (1.69 g, 13.81 mmol) and N,N'-dicyclohexylcarbodiimide (4.27 g, 20.71 mmol) in sequence. Stir the reaction at room temperature for 12 h, and TLC shows that the reaction is complete. Add water (50 ml) and extract with dichloromethane (50 ml x 3). Combine the organic phases, wash with saturated brine (20 ml), dry over anhydrous sodium sulfate, concentrate, and purify by silica gel column chromatography to obtain intermediate A-2-34b (3.2 g, 10.68 mmol, yield 77.4%). 1H NMR (400 MHz, CDC13) δ 5.07 (d, J = 8.8 Hz, 1H), 4.25 (dd, J = 9.1, 4.6 Hz, 1H), 4.04 (qd, J = 10.6, 7.2 Hz, 2H), 2.25 (dd, J = 15.1, 7.5 Hz, 1H), 2.20 - 2.09 (m, 1H), 1.78 (dt, J = 7.7, 2.9 Hz, 2H), 1.70 - 1.53 (m, 4H), 1.47 (s, 9H), 1.33 - 1.20 (m, 2H), 1.02 - 0.97 (m, 3H), 0.92 (d, J = 6.9 Hz, 3H).

[0289] Second Step

[0290] In a dry single neck flask was charged with intermediate A-2-34b (3.2 g, 10.68 mmol) and dichloromethane (20 ml). Then 4M hydrogen chloride solution in dioxane (20 mL) was added and stirred at room temperature for 2 hours. After TLC showed the reaction was complete, the reaction was directly concentrated to get the crude intermediate A-2-34c which was directly used for the next step.

[0291] Third Step

[0292] The crude intermediate A-2-34c and intermediate A-2-7b (1.98 g, 9.94 mmol) were dissolved in dichloromethane (40 ml) and under ice bath, N,N-diisopropylethylamine (3.85 g, 29.82 mmol), N-hydroxy-7-azabenzotriazole (1.35 g, 9.94 mmol), l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (1.91 g, 9.94 mmol) were added. After continued stirring for 1 hour under ice bath, it was allowed to warm to room temperature overnight. After TLC showed the reaction was complete, water (50 ml) was added and extracted with dichloromethane (50 ml x 3). The organic phase was combined and washed with saturated sodium chloride solution (100 ml), dried over anhydrous sodium sulfate and concentrated to give intermediate A-2-34d (2.4 g, 6.27 mmol, 58.7% overall yield for two steps). 1H NMR (400 MHz, CDC13) δ 4.79 - 4.64 (m, 1H), 4.59 (dd, J = 8.9, 4.6 Hz, 1H), 4.17 - 4.02 (m, 2H), 4.01 - 3.92 (m, 1H), 3.85 (dd, J = 14.0, 8.5 Hz, 1H), 2.51 (d, J = 6.2 Hz, 1H), 2.40 (s, 1H), 2.31 - 2.19 (m, 2H), 1.79 (d, J = 10.4 Hz, 2H), 1.61 (ddd, J = 26.9, 13.8, 6.0 Hz, 4H), 1.49 (s, 9H), 1.36 - 1.25 (m, 2H), 1.00 (d, J = 6.8 Hz, 3H), 0.95 (d, J = 6.9 Hz, 3H).

[0293] Fourth Step

[0294] In a dry single neck flask was added intermediate A-2-34d (1.2 g, 3.13 mmol), dichloromethane (5 ml), and 4M hydrogen chloride in dioxane (10 ml) and allowed to react at room temperature for 2 hours. After TLC showed the reaction was complete, it was concentrated to give crude intermediate A-2-34e.

[0295] Fifth Step

[0296] The crude intermediate A-2-34e was dissolved in tetrahydrofuran (20 ml) and N,N-diisopropylethylamine (1.22 g, 9.42 mmol) was added. Acetic anhydride (0.64 g, 6.28 mmol) was added slowly drop wise under ice bath and the reaction was continued for 30 minutes, then it was allowed to warm to room temperature and the reaction was continued for 2 hours. After TLC showed the reaction was complete, ethanol (5 mL) was added, concentrated and purified by silica gel column chromatography to give the product A-2-34 (0.7 g, 2.16 mmol, 68.8% yield). MS-ESI [M+H] Calc. for C2oH29Cl2N5O4: 325.2; found: 325.2. + 1 ​H NMR (400 MHz, CDC13) δ 8.20 (d, J = 8.4 Hz, 1H), 4.85 (dd, J = 9.5, 6.6 Hz, 1H), 4.42 (dd, J = 8.6, 4.7 Hz, 1H), 4.00 (t, J = 6.4 Hz, 2H), 3.98 - 3.87 (m, 2H), 2.67 - 2.51 (m, 1H), 2.44 - 2.28 (m, 1H), 2.20 (d, J = 4.8 Hz, 2H), 1.87 (s, 3H), 1.68 (dd, J = 12.1, 5.0 Hz, 2H), 1.60 - 1.44 (m, 4H), 1.23 - 1.14 (m, 2H), 0.92 (d, J = 6.9 Hz, 3H), 0.87 (d, J = 6.9 Hz, 3H).

[0297] Preparation of compound A-2-35

[0298]

[0299] First step

[0300] Into a dry single necked flask was added compound A-2-34a (3 g, 13.81 mmol) and dichloromethane (50 ml) and stirred in ice bath for 10 min. Then cyclohexanol (2.07 g, 20.71 mmol), 4-dimethylaminopyridine (1.69 g, 13.81 mmol) and N,N'-dicyclohexylcarbodiimide (4.27 g, 20.71 mmol) were added successively. The reaction was continued at room temperature for 12 h. TLC showed the completion of the reaction. Water (50 ml) was added and extracted with dichloromethane (50 ml x 3). The organic phase was washed with saturated sodium chloride solution (100 ml), dried over anhydrous sodium sulphate and purified by silica gel column chromatography to get the intermediate A-2-35a (3.5 g, 11.68 mmol, 84.66% yield). 1 H NMR (400 MHz, CDC13) δ 8.20 (d, J = 8.4 Hz, 1H), 4.85 (dd, J = 9.5, 6.6 Hz, 1H), 4.42 (dd, J = 8.6, 4.7 Hz, 1H), 4.00 (t, J = 6.4 Hz, 2H), 3.98 - 3.87 (m, 2H), 2.67 - 2.51 (m, 1H), 2.44 - 2.28 (m, 1H), 2.20 (d, J = 4.8 Hz, 2H), 1.87 (s, 3H), 1.68 (dd, J = 12.1, 5.0 Hz, 2H), 1.60 - 1.44 (m, 4H), 1.23 - 1.14 (m, 2H), 0.92 (d, J = 6.9 Hz, 3H), 0.87 (d, J = 6.9 Hz, 3H).

[0301] Second step

[0302] In a dry single neck flask was placed intermediate A-2-35a (3.5 g, 11.68 mmol), dichloromethane (20 ml) and 4M hydrogen chloride in dioxane (20 ml). The reaction was allowed to proceed at room temperature for 2 hours. Upon completion of the reaction as indicated by TLC, it was concentrated to obtain crude intermediate A-2-35b.

[0303] Third step

[0304] To a solution of crude intermediate A-2-35b and intermediate A-2-7b (1.98 g, 9.94 mmol) in dichloromethane (40 ml) was added N, N-diisopropylethylamine (3.85 g, 29.82 mmol), N-hydroxy-7-azabenzotriazole (1.35 g, 9.94 mmol), l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (1.91 g, 9.94 mmol) at 0°C under ice bath. The reaction was allowed to proceed at room temperature for 1 hour and then overnight. Upon completion of the reaction as indicated by TLC, it was diluted with water (50 ml) and extracted with dichloromethane (50 ml x 3). The combined organic layers were washed with saturated sodium chloride solution (100 ml), dried over anhydrous sodium sulfate and concentrated. The residue was purified by column chromatography on silica gel to obtain intermediate A-2-35c (2.4 g, 6.27 mmol, 53.7% overall yield for two steps). 1 H NMR (400 MHz, CDC13) δ 4.93 - 4.82 (m, 1H), 4.74 - 4.63 (m, 1H), 4.56 (dd, J = 9.0, 4.6 Hz, 1H), 3.97 (dd, J = 16.3, 8.2 Hz, 1H), 3.90 - 3.79 (m, 1H), 2.45 (d, J = 45.1 Hz, 2H), 2.30 - 2.18 (m, 1H), 1.93 - 1.82 (m, 2H), 1.79 - 1.68 (m, 2H), 1.61 - 1.51 (m, 2H), 1.49 (s, 9H), 1.46 - 1.38 (m, 2H), 1.37 - 1.26 (m, 2H), 1.00 (d, J = 6.9 Hz, 3H), 0.95 (d, J = 6.9 Hz, 3H).

[0305] Fourth step

[0306] In a dry single neck flask was placed intermediate A-2-35c (1.2 g, 3.13 mmol,) dichloromethane (5 ml) and 4M hydrogen chloride in dioxane (10 ml). The reaction was allowed to proceed at room temperature for 2 hours. Upon completion of the reaction as indicated by TLC, it was concentrated to obtain crude intermediate A-2-35d.

[0307] Fifth step

[0308] The crude intermediate A-2-35d was dissolved in tetrahydrofuran (20 ml), N, N- diisopropylethylamine (1.22 g, 9.42 mmol) was added under ice-bath, and acetic anhydride (0.64 g, 6.28 mmol) was added dropwise slowly. After 30 minutes, the reaction was continued at room temperature for 2 hours. TLC showed the reaction was completed, ethanol (5 ml) was added, concentrated, and purified by silica gel column chromatography to give the product A-2-35 (0.7 g, 2.16 mmol, 68.8% yield). MS-ESI [M+H] calculated for C19H29N5O4: 397.2; found: 397.2. + : 325.2; found: 325.2. 1 H NMR (400 MHz, CDC13) δ 8.19 (d, J = 8.4 Hz, 1H), 4.85 (dd, J = 9.5, 6.5 Hz, 1H), 4.80 - 4.69 (m, 1H), 4.41 (dd, J = 8.7, 4.6 Hz, 1H), 4.00 (t, J = 7.7 Hz, 2H), 2.67 - 2.54 (m, 1H), 2.37 (ddt, J = 9.4, 8.4, 7.0 Hz, 1H), 2.25 - 2.11 (m, 1H), 1.87 (s, 3H), 1.81 - 1.71 (m, 2H), 1.69 - 1.57 (m, 2H), 1.49 - 1.23 (m, 6H), 0.92 (d, J = 6.9 Hz, 3H), 0.87 (d, J = 6.9 Hz, 3H).

[0309] Preparation of compound A-2-37

[0310]

[0311] In a dry three-necked flask, nicotinic acid (0.12 g, 0.96 mmol) and 3- (ethylcarbamoylamino)-N,N-dimethylpropan-1-amine hydrochloride (0.18 g, 0.96 mmol), 1-hydroxybenzotriazole (0.13 g, 0.96 mmol) and triethylamine (0.16 g, 1.60 mmol) were charged. Dichloromethane (5 ml) was added, the reaction was stirred for 0.5 hour, and intermediate A-2-21a (0.20 g, 0.80 mmol) was added. After the reaction was carried out at room temperature for 2 hours, water (10 ml) was added, and extraction was performed with ethyl acetate (10 ml x 3). The organic phases were combined, washed with saturated brine (10 ml x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give the product A-2-37 (53.5 mg, 0.17 mmol, 21.0% yield). MS-ESI [M+H] calculated for C21H29N7O4: 460.2; found: 460.3. + : 325.2; found: 325.2. 1H NMR (400 MHz, Chloroform-d) δ 8.88 (s, 1H), 8.74 (d, J = 4.6 Hz, 1H), 8.04 (d, J = 7.8 Hz, 1H), 7.84 (d, J = 8.2 Hz, 1H), 7.44 (dd, J = 7.9, 4.7 Hz, 1H), 5.14 (t, J = 8.0 Hz, 1H), 4.69 - 4.37 (m, 2H), 4.21 (q, J = 8.3 Hz, 1H), 3.75 (s, 3H), 2.88 (s, 1H), 2.49 (s, 1H), 2.20 (d, J = 14.9 Hz, 1H), 0.92 (dd, J = 12.7, 6.8 Hz, 6H).

[0312] Preparation of compound B-2-1

[0313]

[0314] First step

[0315] The starting material B-2-1a (5.00 g, 48.54 mmol) was dissolved in 1,4-dioxane / water (50 mL / 10 mL), and a solution of benzhydryl carbamate succinimidyl ester (12.10 g, 48.54 mmol) in 1,4-dioxane / water (50 mL / 10 mL) was slowly added thereto. The reaction liquid was reacted at 25°C for 16 hours. After the reaction was completed, ethyl acetate was added for dilution (200 mL), and then washed with 5% sodium bicarbonate solution and 5% citric acid solution, respectively. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain the crude intermediate B-2-1b (10.40 g, 43.83 mmol, yield 90.3%). MS-ESI [M+H] calc’d for C17H20N4O5: 238.2; found: 238.1. +

[0316] Second step

[0317] The intermediate B-2-1b (10.40 g, 43.83 mmol) was dissolved in ethyl acetate (300 mL), and Dess-Martin oxidizing agent (22.30 g, 52.60 mmol) was added thereto. The reaction liquid was stirred at room temperature for 24 hours. After the reaction was completed, the reaction liquid was filtered with celite, and the filtrate was concentrated and passed through a silica gel column to obtain the intermediate B-2-1c (8.76 g, 37.25 mmol, yield 85%). MS-ESI [M+H] calc’d for C16H18N4O5: 236.1; found: 236.1. +

[0318] Third step

[0319] ​​Intermediate B-2-1c (7.0 g, 29.79 mmol) and glyoxal solution (15.4 mL, 119.16 mmol) were dissolved in methanol (70 mL), and ammonia water (18.9 mL, 148.95 mmol) was slowly added thereto while maintaining the temperature below 10 °C during the dropwise addition. The reaction solution was stirred at 25 °C for 16 hours. After the completion of the reaction, the reaction solution was poured into ice water, and the precipitated solid was collected by filtration. The obtained filter cake was washed with water, and intermediate B-1-2d (6.3 g, 23.05 mmol, yield 77%) was obtained by drying. MS-ESI [M+H] calc’d for C8H10N2O2, 174.1; found, 174.1. + : 174.1; found, 174.1.

[0320] Fourth step

[0321] Intermediate B-2-1d (3.00 g, 10.98 mmol) was dissolved in methanol (75 mL), followed by the addition of palladium on carbon (0.3 g, 10 wt.%). The mixture was purged with hydrogen gas three times, and the reaction solution was reacted under a hydrogen atmosphere at 25 °C for 16 hours. After the completion of the reaction, the reaction solution was filtered with celite, and the filtrate was concentrated to obtain crude intermediate B-2-1e (1.2 g, 8.62 mmol, yield 78%). MS-ESI [M+H] calc’d for C8H10N2O2, 140.0; found, 140.1. + : 140.0; found, 140.1.

[0322] Fifth step

[0323] Intermediate B-2-1e (1.2 g, 8.62 mmol) and A-2-1b (1.82 g, 9.05 mmol) were dissolved in N,N-dimethylformamide (12 mL), and 1-hydroxybenzotriazole (1.40 g, 10.34 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.97 g, 10.34 mmol) were added at -20 °C. The reaction solution was stirred at 0 °C for 3 hours. The reaction solution was diluted with water, and extracted with ethyl acetate three times. The combined organic phase was washed with water and saturated brine, dried with anhydrous sodium sulfate, and concentrated. The obtained residue was separated and purified with a silica gel column (dichloromethane / methanol = 15 / 1), and then purified by reverse phase preparative purification to obtain intermediate B-2-1f (0.90 g, 2.80 mmol, yield 32%). MS-ESI [M+H] calc’d for C21H22N4O2, 323.2; found, 323.1. + : 323.2; found, 323.1.

[0324] Sixth step

[0325] Intermediate B-2-1f (0.90 g, 2.80 mmol) was dissolved in ethyl acetate (20 mL), hydrogen chloride in dioxane (4.0 M, 3 ml) was added dropwise at room temperature, and the mixture was stirred at room temperature for 12 hours after the dropwise addition was completed. The reaction was monitored to completion, and the residue was concentrated to dryness to obtain crude intermediate B-2-1g (0.60 g, 2.33 mmol, yield 86%). MS-ESI [M+H] calc’d for C9H13N5O2, 223.1; found, 223.2. + : 223.1; found, 223.2.

[0326] Seventh step

[0327] Intermediate B-2-1g (0.60 g, 2.33 mmol) was dissolved in toluene (10 mL), and acetic anhydride (2.38 g, 23.30 mmol) was added. The mixture was heated to 110 °C, and the reaction was allowed to proceed for 8 hours. The reaction was monitored to completion, and the residue was concentrated to dryness. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 15 / 1) and then by reverse phase preparative purification to obtain product B-2-1 (0.20 g, 0.76 mmol, yield 33%). MS-ESI [M+H] calc’d for C9H13N5O2, 223.1; found, 223.2. + : 223.1; found, 223.2. 1 H NMR (400 MHz, DMSO) δ 11.84 (d, J = 42.0 Hz, 1H), 8.58 - 8.11 (m, 1H), 7.02 - 6.71 (m, 2H), 4.95 - 4.62 (m, 2H), 4.03 (t, J = 7.7 Hz, 1H), 3.81 - 3.68 (m, 1H), 2.44 - 2.26 (m, 1H), 2.20 - 1.90 (m, 2H), 1.83 - 1.52 (m, 3H), 0.84 (ddd, J = 10.3, 8.3, 5.0 Hz, 3H), 0.75 (t, J = 6.7 Hz, 3H).

[0328] Preparation of compound B-2-2

[0329]

[0330] First step

[0331] Compound B-2-2a (15 g, 0.21 mol) was dissolved in dichloromethane (250 mL), and (R)-tert-butylsulfinamide (27.5 g, 0.23 mol) and anhydrous copper sulfate (8.6 g, 0.05 mol) were added. The resulting reaction system was stirred at 25 °C for 24 hours, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain intermediate B-2-2b (7 g, 39.3 mmol, yield 17.6%). MS-ESI [M+H] calc’d for C9H13N5O2, 223.1; found, 223.2. +: 176.1 ; Found: 176.1.

[0332] Second Step

[0333] Compound 1, 3-oxazole (1 g, 14.5 mmol) was dissolved in tetrahydrofuran (30 mL), after nitrogen replacement, borane tetrahydrofuran solution (22 mL) was added, the reaction solution was reduced to -78°C, then n-butyllithium tetrahydrofuran solution (2.4 M, 10 mL, 24 mmol) was added, the reaction was stirred for 30 minutes, then intermediate B-2-2b (2.5 g, 14.5 mmol) was slowly added, the reaction solution was stirred at -78°C for 3 hours. After the reaction was quenched with saturated aqueous ammonium chloride solution, it was extracted with ethyl acetate three times (50 mL x 3), the organic phase was combined and washed with saturated sodium bicarbonate solution (100 mL x 2), saturated ammonium chloride solution (100 mL x 2) and saturated brine (100 mL x 2) in turn. The obtained organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, then purified by silica gel column chromatography (dichloromethane / methanol = 10:1) to obtain intermediate B-2-2c (1 g, 4.1 mmol, yield 28.7%). MS-ESI Theoretical value [M+H] + : 245.1 ; Found: 245.1.

[0334] Third Step

[0335] Intermediate B-2-2c (1 g, 4.1 mmol) was added to 2M hydrogen chloride solution in 1,4-dioxane (10 mL) at 0°C, the reaction was stirred at 0°C for 1 hour, then concentrated to obtain intermediate B-2-2d (400 mg, 2.85 mmol, yield 69.7%). MS-ESI Theoretical value [M+H] + : 141.1 ; Found: 141.1.

[0336] Fourth Step

[0337] To a solution of intermediate A-2-6b (2.5 g, 12.4 mmol) in dichloromethane (30 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.57 g, 18.6 mmol), 1-hydroxybenzotriazole (2.51 g, 18.6 mmol), N-methylmorpholine (11.29 g, 111.6 mmol) were added, and after stirring at -15 °C for 10 min, intermediate B-2-2d (1.91 g, 13.64 mmol) was added. The reaction was stirred at -15 °C for 4 h. After the reaction was completed, the reaction was washed with saturated citric acid solution (50 mL x 2), saturated sodium carbonate solution (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The obtained crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give intermediate B-2-2e (2.5 g, 7.7 mmol, 56.7% yield). MS-ESI [M+H] calc’d for C19H22F2N4O2: 324.2; found: 324.2. + : 324.2; found: 324.2.

[0338] Fifth step

[0339] Intermediate B-2-2e (2.5 g, 6.4 mmol) was dissolved in a solution of trifluoroacetic acid in dichloromethane (V / V = 1:1, 40 mL), and the reaction was stirred at room temperature for 30 min. After the reaction was completed, it was concentrated to give the crude intermediate B-2-2f (5.58 g). MS-ESI [M+H] calc’d for C19H22F2N4O2: 324.2; found: 324.2. + : 324.2; found: 324.2.

[0340] Sixth step

[0341] To a solution of the crude intermediate B-2-2f (5.58 g) in dichloromethane (40 mL), N,N-diisopropylethylamine (7.96 g, 61.6 mmol) was added, and the reaction was purged with nitrogen three times, stirred at 0 °C for 5 min, and then deuterated acetyl chloride (0.69 g, 8.4 mmol) was added. The reaction was stirred at room temperature for 2 h. The reaction was washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The product B-2-2 (1.57 g, 5.85 mmol, 75.6% total yield over two steps) was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1). MS-ESI [M+H] calc’d for C19H22F2N4O2: 324.2; found: 324.2. + : 324.2; found: 324.2. 1H NMR (400 MHz, d6-DMSO) δ 8.64 (dd, J = 64.7, 8.7 Hz, 1H), 8.02 (dd, J = 7.4, 0.6 Hz, 1H), 7.13 (d, J = 7.8 Hz, 1H), 4.92 - 4.61 (m, 2H), 4.00 (t, J = 7.6 Hz, 1H), 3.71 (dt, J = 8.8, 6.3 Hz, 1H), 2.44 - 2.23 (m, 1H), 2.23 - 1.85 (m, 2H), 0.86 (dd, J = 11.2, 6.8 Hz, 3H), 0.77 (dd, J = 11.0, 6.8 Hz, 3H).

[0342] Preparation of compound B-2-3

[0343]

[0344] First step:

[0345] To a solution of compound A-2-7b (2 g, 9.96 mmol) in dichloromethane (50 mL) was added l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.39 g, 12.45 mmol), 1-hydroxybenzotriazole (1.68 g, 12.45 mmol), N,N-diisopropylethylamine (5.36 g, 41.50 mmol) at -10 °C. After stirring for 10 min, intermediate B-2-2d (1.47 g, 8.30 mmol) was added and the reaction was continued at -10 °C for 2 h. After the reaction was completed, the reaction solution was washed with saturated aqueous citric acid solution (50 mL x 2), saturated sodium carbonate solution (50 mL x 2) successively. The obtained organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (dichloromethane / methanol = 50:1) to give intermediate B-2-3a (2 g, 6.2 mmol, 62.2%). MS-ESI Calcd for [M+H] + : 324.2; Found: 324.0.

[0346] Second step:

[0347] Intermediate B-2-3a (2 g, 6.2 mmol) was dissolved in trifluoroacetic acid in dichloromethane solution (V / V = 1:1, 20 mL), and the reaction solution was stirred at room temperature for 2 h. After the reaction was completed, it was concentrated to give intermediate B-2-3b crude (3.6 g). MS-ESI Calcd for [M+H] + : 224.1; Found: 224.0.

[0348] Third step:

[0349] To a solution of intermediate B-2-3b crude (3.6 g) in dichloromethane (60 mL) was added N,N-diisopropyl ethylamine (6.4 g, 49.6 mmol), deuterated acetyl chloride (606 mg, 7.4 mmol) sequentially at 0 °C. After addition, the reaction was continued at 0 °C for 1 h. After the reaction was completed, the reaction solution was concentrated and purified by silica gel column chromatography (dichloromethane / methanol = 30 / 1) to give product B-2-3 (1.22 g, 4.55 mmol, 73.5% overall yield for two steps). MS-ESI Calcd for [M+H] + : 269.2; Found: 269.0. 1H NMR (400 MHz, Chloroform-d) δ 8.50 (d, J = 8.5 Hz, 1H), 7.60 (s, 1H), 7.08 (s, 1H), 5.08 (dd, J = 8.5, 6.0 Hz, 1H), 4.95 (dd, J = 9.6, 6.6 Hz, 1H), 4.06 (t, J = 8.0 Hz, 2H), 2.73 - 2.59 (m, 1H), 2.51 - 2.35 (m, 1H), 2.28 (dq, J = 13.4, 6.7 Hz, 1H), 0.96 (dd, J = 13.9, 6.8 Hz, 6H).

[0350] Preparation of compound B-2-4

[0351]

[0352] First step

[0353] Intermediate B-2-3a (6 g, 18.55 mmol) was dissolved in dichloromethane (50 mL), and 4 M hydrogen chloride solution in dioxane (15 mL) was added. The reaction was continued at room temperature for 2 h, and TLC showed that the reaction was complete. Concentration gave intermediate B-2-4a crude (4.83 g, 18.6 mmol, yield 100%). MS-ESI Calcd for [M+H] + : 224.1; Found: 224.3.

[0354] Second step

[0355] In a dry single-mouth flask, intermediate B-2-4a (500.0 mg, 1.93 mmol) was dissolved with super-dry dichloromethane (5 mL) and replaced with nitrogen. Then, diisopropyl ethyl amine (0.75 g, 5.79 mmol) was added at 0 °C, stirred for 10 min, and finally, cyclopropyl formyl chloride (262.3 mg, 2.51 mmol) was slowly added to the above reaction system. The reaction was continued at 0 °C for 2 h, TLC showed that the reaction was complete, water (10 mL) was added, extracted and separated, then the aqueous phase was continuously extracted with dichloromethane (10 mL x 2). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (EA: PE = 10% ~ 50%), to obtain the product B-2-4 (351.0 mg, 1.20 mmol, yield 62.40%). MS-ESI [M+H] Theoretical value: 292.2; Found: 292.1. + 1 H NMR (400 MHz, Chloroform-d) δ 8.37 (d, J = 8.2 Hz, 1H), 7.52 (s, 1H), 6.99 (s, 1H), 4.99 (dd, J = 8.5, 6.3 Hz, 1H), 4.87 (dd, J = 9.4, 6.7 Hz, 1H), 4.27 - 3.93 (m, 2H), 2.72 - 2.53 (m, 1H), 2.48 - 2.36 (m, 1H), 2.22 - 2.13 (m, 1H), 1.41 - 1.33 (m, 1H), 1.01 - 0.90 (m, 2H), 0.86 (dd, J = 13.0, 6.8 Hz, 6H), 0.81 - 0.73 (m, 2H).

[0356] Preparation of compound B-2-5

[0357]

[0358] ​In a dry single-neck flask, intermediate B-2-4a (500.0 mg, 1.93 mmol) was dissolved with super dry dichloromethane (5 mL) and replaced with nitrogen. Then, diisopropyl ethyl amine (0.75 g, 5.79 mmol) was added at 0 °C, stirred for 10 min, and finally, trimethyl acetyl chloride (302.5 mg, 2.51 mmol) was slowly added to the above reaction system. The reaction was continued at 0 °C for 2 h, TLC showed that the reaction was complete, water (10 mL) was added, extracted and separated, and then the aqueous phase was continuously extracted with dichloromethane (10 mL x 2). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography (EA: PE = 10% ~ 50%) to obtain the product B-2-5 (170.0 mg, 0.55 mmol, yield 28.7%). MS-ESI [M+H] Theoretical value: 308.2; Found: 308.2. + 1 H NMR (400 MHz, Chloroform-d) δ 8.17 (d, J = 8.4 Hz, 1H), 7.50 (s, 1H), 6.98 (s, 1H), 5.00 (dd, J = 8.9, 6.1 Hz, 1H), 4.90 (dd, J = 9.6, 5.9 Hz, 1H), 4.24 (t, J = 7.7 Hz, 2H), 2.59 (dq, J = 14.1, 7.2 Hz, 1H), 2.44 - 232 (m, 1H), 2.27 - 2.09 (m, 1H), 1.14 (s, 9H), 0.87 (dd, J = 11.6, 6.8 Hz, 6H).

[0359] Preparation of compound B-2-8

[0360]

[0361] First step

[0362] B-2-8a (5 g, 46.68 mmol) was dissolved in super dry dichloromethane (100 ml), and cesium carbonate (18.25 g, 56.02 mmol) and (S)-tert-butylsulfinamide (5.66 g, 46.68 mmol) were added in turn. Stirring at room temperature for 2 h, TLC showed that the reaction was complete, water (100 ml) was added, extracted with dichloromethane (50 ml x 2). The combined organic phase was washed with saturated sodium chloride solution (200 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain intermediate B-2-8b (9.56 g, 45.52 mmol, yield 97.4%).

[0363] Second step

[0364] ​Intermediate B-2-8b (9.56 g, 45.46 mmol) was dissolved in super dry tetrahydrofuran (100 ml) under nitrogen protection and cooled to -78 °C. 1M isopropyl magnesium bromide (50 ml) was added dropwise slowly. After the addition was completed, the reaction was continued for 2 h. TLC showed that the reaction was complete, water (100 ml) was added, extracted with dichloromethane (50 ml x 2). The organic phase was combined, washed with saturated sodium chloride solution (200 ml), dried over anhydrous sodium sulfate and purified by silica gel column chromatography to obtain intermediate B-2-8c (6.15 g, 24.21 mmol, yield 53.2%). + : 255.2; found: 255.3.

[0365] Third step

[0366] Intermediate B-2-8c (6.15 g, 24.21 mmol) was dissolved in dichloromethane (100 ml), and 4M hydrogen chloride solution in dioxane (30 ml) was added. The reaction was continued at room temperature for 2 hours, TLC showed that the reaction was complete, concentrated to obtain intermediate B-2-8d crude product.

[0367] Fourth step

[0368] Intermediate B-2-8d crude product (0.5 g, 2.48 mmol) was dissolved in dichloromethane (50 ml), and intermediate A-2-7b (0.56 g, 3.72 mmol), N, N-diisopropyl ethylamine (0.96 g, 7.44 mmol) and 1-hydroxybenzotriazole (0.5 g, 3.72 mmol) were added in turn under ice bath. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.71 g, 3.72 mmol) was added and the reaction was continued for 30 minutes under ice bath, and then the temperature was raised to room temperature. After 3 hours of reaction, TLC showed that the raw material was completely reacted. Water (50 ml) was added, extracted with dichloromethane (50 ml x 3), washed with saturated brine (100 ml), dried over anhydrous sodium sulfate and purified by silica gel column chromatography to obtain intermediate B-2-8e (0.60 g, 1.80 mmol, total yield of two steps 72.4%). 1H NMR (400 MHz, CDC13) δ 8.56 (t, J = 5.3 Hz, 1H), 7.85 (s, 1H), 7.63 (td, J = 7.7, 1.6 Hz, 1H), 7.19 (t, J = 8.5 Hz, 2H), 4.91 (dd, J = 8.9, 7.0 Hz, 1H), 4.77 - 4.59 (m, 1H), 3.92 (dd, J = 16.3, 8.3 Hz, 1H), 3.79 (td, J = 8.6, 5.3 Hz, 1H), 2.46 (s, 1H), 2.22 (dd, J = 12.7, 7.1 Hz, 2H), 1.50 (s, 9H), 0.94 (d, J = 6.8 Hz, 3H), 0.84 (d, J = 6.7 Hz, 3H).

[0369] Fifth Step

[0370] Intermediate B-2-8e (0.60 g, 1.8 mmol) was dissolved in dichloromethane (5 mL) and 4M hydrogen chloride in dioxane (8 mL) was added. The reaction was stirred at room temperature for 2 hours, TLC showed the reaction was complete, concentrated to give intermediate B-2-8f (0.42 g, 1.8 mmol, yield 100%). 1 H NMR (400 MHz, MeOD) δ 8.81 (d, J = 5.4 Hz, 1H), 8.65 (t, J = 7.5 Hz, 1H), 8.13 (d, J = 8.2 Hz, 1H), 8.08 - 7.93 (m, 1H), 5.26 - 5.14 (m, 1H), 4.95 (d, J = 7.8 Hz, 1H), 4.19 - 4.05 (m, 1H), 3.95 (td, J = 10.0, 6.3 Hz, 1H), 2.94 (dtd, J = 15.8, 9.5, 6.3 Hz, 1H), 2.58 (dq, J = 12.2, 7.8 Hz, 1H), 2.33 (dd, J = 13.9, 6.8 Hz, 1H), 1.11 (dd, J = 11.6, 4.3 Hz, 3H), 0.90 (dd, J = 12.5, 6.8 Hz, 3H).

[0371] Sixth Step

[0372] Intermediate B-2-8f (0.42 g, 1.8 mmol) was dissolved in tetrahydrofuran (20 mL). N,N-diisopropylethylamine (0.70 g, 5.40 mmol) was added, and acetic anhydride (0.37 g, 3.60 mmol) was added dropwise slowly under ice bath. The reaction was continued for 30 minutes under ice bath, then was removed to room temperature. The reaction was stirred for 2 hours, TLC showed the reaction was complete. Methanol (5 mL) was added, concentrated, and purified by silica gel column chromatography to give product B-2-8 (0.25 g, 0.91 mmol, yield 50.4%). MS-ESI [M+H]+ : 276.2; found: 276.3. 1 HNMR (400 MHz, CDC13) δ 8.49 (d, J = 4.5 Hz, 1H), 8.27 (d, J = 8.1 Hz, 1H), 7.68 - 7.51 (m, 1H), 7.19 - 7.05 (m, 2H), 4.86 (dd, J = 8.7, 6.0 Hz, 1H), 4.83 - 4.74 (m, 1H), 4.06 - 3.91 (m, 2H), 2.58 - 2.44 (m, 1H), 2.37 (ddd, J = 19.4, 11.8, 7.9 Hz, 1H), 2.17 (dd, J = 13.5, 6.8 Hz, 1H), 1.91 - 1.83 (m, 3H), 0.85 (t, J = 6.9 Hz, 3H), 0.81 - 0.73 (m, 3H).

[0373] Preparation of compounds B-2-9 and B-2-10

[0374]

[0375] First step

[0376] B-2-9a (2 g, 18.67 mmol) was dissolved in super dry dichloromethane (100 ml), cesium carbonate (7.30 g, 22.40 mmol) and (S)-tert-butylsulfinamide (2.26 g, 18.67 mmol) were added successively. Stirring at room temperature for 2 hours. TLC showed that the reaction was completed, water (100 ml) was added, extracted with dichloromethane (50 ml x 2). The combined organic phase was washed with saturated sodium chloride solution (200 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain intermediate B-2-9b (3.82 g, 18.02 mmol, yield 97.38%).

[0377] Second step

[0378] Intermediate B-2-9b (3.82 g, 18.02 mmol) was dissolved in super dry tetrahydrofuran (100 ml), nitrogen was replaced and cooled to -78°C, 1M isopropyl magnesium bromide tetrahydrofuran solution (20 ml) was added slowly dropwise. After the addition was completed, the reaction was continued at -78°C for 2 hours. TLC showed that the reaction was completed, water (100 ml) was added, extracted with dichloromethane (50 ml x 2). The combined organic phase was washed with saturated sodium chloride solution (200 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain two components.

[0379] The component 1 with shorter retention time was intermediate B-2-9c (1.88 g, 7.43 mmol, yield 41.26%), MS-ESI [M+H]+ : 255.2, found: 255.3.

[0380] The component 2 with long retention time was B-2-10c (1.83 g, 7.22 mmol, yield 40.10%), MS-ESI [M+H] calculated C26H32N4O4 488.2, found 488.2. + : 255.2, found: 255.3.

[0381] Step 3 - Component 1

[0382] The intermediate B-2-9c (1 g, 3.93 mmol) was dissolved in dichloromethane (10 ml), and 4M hydrogen chloride solution in dioxane (5 ml) was added. The reaction was carried out at room temperature for 3 hours, and TLC detection showed that the reaction was complete. Concentration gave the crude intermediate B-2-9d.

[0383] Step 4 - Component 1

[0384] The crude intermediate B-2-9d was dissolved in dichloromethane (30 ml), and N,N- diisopropylethylamine (1.02 g, 7.86 mmol) was added, followed by the addition of intermediate A-2-7b (0.87 g, 4.32 mmol), 1-hydroxybenzotriazole (0.53 g, 3.93 mmol) and 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.75 g, 3.93 mmol). The reaction was carried out at room temperature overnight, water (50 ml) was added, and extraction was carried out with dichloromethane (50 ml x 3). The combined organic phases were washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, and separated by silica gel column chromatography to give intermediate B-2-9e (0.8 g, 2.40 mmol, total yield 61.1% for two steps). 1 HNMR (400 MHz, CDC13) δ 8.48 (d, J = 5.9 Hz, 2H), 7.14 (d, J = 5.9 Hz, 2H), 4.71 (dd, J = 8.4, 6.3 Hz, 1H), 4.62 (d, J = 7.2 Hz, 1H), 3.88 (dd, J = 16.3, 8.1 Hz, 1H), 3.76 (tt, J = 16.3, 8.2 Hz, 1H), 2.39 (d, J = 51.6 Hz, 2H), 2.02 (td, J = 13.2, 6.6 Hz, 1H), 1.40 (s, 9H), 0.90 - 0.67 (m, 6H).

[0385] Step 5 - Component 1

[0386] The intermediate B-2-9e (0.8 g, 2.4 mmol) was dissolved in dichloromethane (5 ml), and 4M hydrogen chloride solution in dioxane (8 ml) was added. The reaction was carried out at room temperature for 2 hours, and the reaction was complete. Concentration gave the crude intermediate B-2-9f.

[0387] Step 6 - Component 1

[0388] The crude intermediate B-2-9f was dissolved in tetrahydrofuran (20 ml), and N,N-diisopropylethylamine (1.02 g, 7.86 mmol) was added. Acetic anhydride (0.6 g, 5.9 mmol) was added dropwise slowly under ice bath. The reaction was continued for 30 minutes, and then was removed to room temperature. After 2 hours of reaction, TLC showed that the reaction was completed. Methanol (2 mL) was added, and then was concentrated. The product B-2-9 (0.30 g, 1.09 mmol, 45.4% total yield over two steps) was isolated by silica gel column chromatography. 1 H NMR (400 MHz, CDC13) δ 8.61 (d, J = 8.2 Hz, 1H), 8.48 (d, J = 5.8 Hz, 2H), 7.15 (d, J = 6.0 Hz, 2H), 4.83 (dd, J = 9.4, 6.6 Hz, 1H), 4.66 (dd, J = 8.2, 6.4 Hz, 1H), 4.03 (t, J = 7.7 Hz, 2H), 2.74 - 2.58 (m, 1H), 2.33 (ddt, J = 12.1, 9.5, 7.4 Hz, 1H), 1.96 (dd, J = 13.4, 6.7 Hz, 1H), 1.91 (s, 3H), 0.81 (d, J = 2.6 Hz, 3H), 0.79 (d, J = 2.7 Hz, 3H).

[0389] Step 3 - Component 2

[0390] The intermediate B-2-10c (1 g, 3.93 mmol) was dissolved in dichloromethane (10 ml), and 4M hydrogen chloride solution in dioxane (5 ml) was added. The reaction was continued at room temperature for 3 hours. TLC detection showed that the reaction was completed, and then was concentrated to give the crude intermediate B-2-10d.

[0391] Step 4 - Component 2

[0392] The crude intermediate B-2-10d was dissolved in dichloromethane (30 ml), and N,N-diisopropylethylamine (1.02 g, 7.86 mmol) was added. Then, the intermediate A-2-7b (0.87 g, 4.32 mmol), 1-hydroxybenzotriazole (0.53 g, 3.93 mmol) and l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.75 g, 3.93 mmol) were added in sequence. The reaction was continued at room temperature overnight. Water (50 ml) was added, and then was extracted with dichloromethane (50 ml x 3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and then was isolated by silica gel column chromatography to give the intermediate B-2-10e (0.8 g, 2.40 mmol, 61.1% total yield over two steps). 1H NMR (400 MHz, CDC13) δ 8.47 (d, J = 5.9 Hz, 2H), 7.09 (d, J = 6.0 Hz, 2H), 4.75 (dd, J = 8.9, 6.4 Hz, 1H), 4.69 (t, J = 7.5 Hz, 1H), 3.85 (dd, J = 16.4, 8.3 Hz, 1H), 3.72 (dd, J = 14.1, 8.4 Hz, 1H), 2.51 - 2.23 (m, 2H), 2.15 (s, 1H), 1.43 (s, 9H), 0.84 (dd, J = 6.7, 4.1 Hz, 6H).

[0393] Fifth Step - Component 2

[0394] Intermediate B-2-10e (0.8 g, 2.4 mmol) was dissolved in dichloromethane (5 mL) and 4 M hydrogen chloride in dioxane (8 mL) was added. The reaction was stirred at room temperature for 2 hours, and was complete. The reaction was concentrated to give crude intermediate B-2-10f.

[0395] Sixth Step - Component 2

[0396] Crude intermediate B-2-10f was dissolved in tetrahydrofuran (20 mL) and N,N- diisopropylethylamine (1.02 g, 7.86 mmol) was added. Acetic anhydride (0.6 g, 5.9 mmol) was added dropwise slowly under ice bath. The reaction was stirred for 30 minutes and was allowed to warm to room temperature. After 2 hours, TLC showed the reaction was complete. Methanol (2 mL) was added and the reaction was concentrated. The product, B-2-10 (0.33 g, 1.2 mmol, 50.0% overall yield over two steps), was isolated by silica gel column chromatography. 1 H NMR (400 MHz, CDC13) δ 8.47 (d, J = 5.9 Hz, 2H), 7.09 (d, J = 6.0 Hz, 2H), 4.75 (dd, J = 8.9, 6.4 Hz, 1H), 4.69 (t, J = 7.5 Hz, 1H), 3.85 (dd, J = 16.4, 8.3 Hz, 1H), 3.72 (dd, J = 14.1, 8.4 Hz, 1H), 2.51 - 2.23 (m, 2H), 2.15 (s, 1H), 1.43 (s, 9H), 0.84 (dd, J = 6.7, 4.1 Hz, 6H).

[0397] Preparation of Compound B-2-11

[0398]

[0399] First Step

[0400] B-2-11a (1 g, 9.33 mmol) was dissolved in super dry dichloromethane (100 ml), and cesium carbonate (3.65 g, 11.20 mmol) and (S)-tert-butylsulfonamide (1.13 g, 9.33 mmol) were added successively. The reaction was carried out at room temperature for 2 hours, and TLC showed that the reaction was completed. Water (100 ml) was added, and extraction was carried out with dichloromethane (50 ml x 2). The organic phase was combined, washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain intermediate B-2-11b (1.90 g, 9.01 mmol, yield 97.3%).

[0401] Second step

[0402] Intermediate B-2-11b (1.90 g, 9.01 mmol) was dissolved in super dry tetrahydrofuran (100 ml), and 1M isopropyl magnesium bromide (10 ml) was slowly added dropwise under nitrogen protection while being cooled to -78°C. After the reaction was continued at this temperature for 2 hours, TLC showed that the reaction was completed. Water (100 ml) was added, and extraction was carried out with dichloromethane (50 ml x 2). The organic phase was combined, washed with saturated sodium chloride solution (200 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain two single configuration components.

[0403] The first component, i.e., intermediate B-2-11c (0.94 g, 3.54 mmol, yield 41.26%), had a short retention time. MS-ESI Theoretical value [M+H] + : 255.2; Found: 255.3.

[0404] The second component, i.e., intermediate B-2-11c component 2 (0.47 g, 1.85 mmol, yield 20.10%), had a long retention time. MS-ESI Theoretical value [M+H] + : 255.2; Found: 255.3.

[0405] Third step

[0406] Intermediate B-2-11c (0.94 g, 3.54 mmol) was dissolved in dichloromethane (10 ml), and 4M hydrogen chloride solution in dioxane (5 ml) was added. The reaction was carried out at room temperature for 3 hours, and TLC showed that the reaction was completed. Concentration was performed to obtain crude intermediate B-2-11d.

[0407] Fourth step

[0408] The crude intermediate B-2-11d was dissolved in dichloromethane (30 ml), N,N-diisopropylethylamine (1.02 g, 7.86 mmol), intermediate A-2-7b (0.87 g, 4.32 mmol), 1-hydroxybenzotriazole (0.53 g, 3.93 mmol) and l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.75 g, 3.93 mmol) were added successively. After the reaction at room temperature overnight, water (50 ml) was added and extracted with dichloromethane (50 ml x 3). The organic phase was combined and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate and purified by silica gel column chromatography to give intermediate B-2-11e (0.30 g, 0.9 mmol, 25.4% overall yield for two steps). 1 H NMR (400 MHz, CDC13) δ 8.60 - 8.47 (m, 2H), 7.59 (dt, J = 7.8, 1.7 Hz, 1H), 7.32 - 7.24 (m, 1H), 4.86 (dd, J = 8.7, 7.0 Hz, 1H), 4.77 (t, J = 7.5 Hz, 1H), 3.93 (q, J = 8.2 Hz, 1H), 3.84 - 3.72 (m, 1H), 2.41 (s, 2H), 2.14 - 2.02 (m, 1H), 1.52 (s, 9H), 0.98 (d, J = 6.8 Hz, 3H), 0.93 (d, J = 6.7 Hz, 3H).

[0409] Fifth Step

[0410] The crude intermediate B-2-11d was dissolved in dichloromethane (30 ml), N,N-diisopropylethylamine (1.02 g, 7.86 mmol), intermediate A-2-7b (0.87 g, 4.32 mmol), 1-hydroxybenzotriazole (0.53 g, 3.93 mmol) and l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.75 g, 3.93 mmol) were added successively. After the reaction at room temperature overnight, water (50 ml) was added and extracted with dichloromethane (50 ml x 3). The organic phase was combined and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate and purified by silica gel column chromatography to give intermediate B-2-11e (0.30 g, 0.9 mmol, 25.4% overall yield for two steps).

[0411] Sixth Step

[0412] The crude intermediate B-2-11d was dissolved in dichloromethane (30 ml), N,N-diisopropylethylamine (1.02 g, 7.86 mmol), intermediate A-2-7b (0.87 g, 4.32 mmol), 1-hydroxybenzotriazole (0.53 g, 3.93 mmol) and l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.75 g, 3.93 mmol) were added successively. After the reaction at room temperature overnight, water (50 ml) was added and extracted with dichloromethane (50 ml x 3). The organic phase was combined and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate and purified by silica gel column chromatography to give intermediate B-2-11e (0.30 g, 0.9 mmol, 25.4% overall yield for two steps). 1H NMR (400 MHz, CDC13) δ 8.73 (d, J = 8.1 Hz, 1H), 8.54 - 8.39 (m, 2H), 7.55 (dt, J = 7.9, 1.8 Hz, 1H), 7.29 - 7.26 (m, 1H), 4.96 (dd, J = 9.4, 6.7 Hz, 1H), 4.79 (dd, J = 8.4, 6.9 Hz, 1H), 4.15 - 3.90 (m, 2H), 2.69 - 2.60 (m, 1H), 2.43 - 2.35 (m, 1H), 2.06 - 2.00 (m, 1H), 1.94 (s, 3H), 0.94 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.8 Hz, 3H).

[0413] Preparation of compound ZZL-7 of Preparation Example 42

[0414]

[0415] First step

[0416] In a dry single neck flask, (tert-butoxycarbonyl)-L-alanine (4.5 g, 23.78 mmol) was dissolved in dichloromethane (50 mL) and stirred in an ice bath for 10 min. Then L-valine methyl ester (3.12 g, 23.78 mmol), N,N-diisopropylethylamine (6.15 g, 47.56 mmol), 1-hydroxybenzotriazole (3.53 g, 26.16 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.01 g, 26.16 mmol) were added to the flask in sequence and stirred at room temperature for 2 h. After TLC showed that the reaction was completed, water (50 mL) was added to quench and adjusted to be acidic with 2M hydrochloric acid, extracted with dichloromethane (50 mL*2), the combined organic phase was adjusted to be basic with saturated sodium bicarbonate solution, extracted with dichloromethane (50 mL*2), washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 2:1) to obtain intermediate (tert-butoxycarbonyl)-L-alanyl-L-valine methyl ester (6.5 g, 21.50 mmol, yield 90.39%).

[0417] LCMS (M+H + ) = 303.1

[0418] Second step

[0419] In a dry single neck flask, (tert-butoxycarbonyl)-L-alanyl-L-valine methyl ester (6.5 g, 21.50 mmol) was dissolved in dichloromethane (30 mL) and then hydrochloric acid-1.4-dioxane (10 mL, 4M) was added to the flask and stirred at room temperature for 2 h. After TLC showed the completion of the reaction, the organic phase was concentrated and purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain intermediate L-alanyl-L-valine methyl ester (3.7 g, 18.32 mmol, 87.22% yield).

[0420] LCMS (M+H + ) = 203.3

[0421] 1 HNMR (400 MHz, CDC13) δ 8.13 (s, 3H), 4.56 (s, 1H), 4.31 (s, 1H), 3.65 (s, 3H), 2.15 (d, J = 5.3 Hz, 1H), 1.57 (s, 3H), 0.92 (t, J = 6.8 Hz, 6H).

[0422] Third step

[0423] Compound L-alanyl-L-valine methyl ester (3.7 g, 18.32 mmol) and N,N- diisopropylethylamine (4.72 g, 36.64 mmol) were dissolved in tetrahydrofuran (50 mL) and stirred at 0 °C after nitrogen replacement, acetic anhydride (2.8 g, 27.48 mmol) was added slowly dropwise after 10 min, stirred at this temperature for 10 min, then moved to room temperature and stirred for 2 h. After TLC showed the completion of the reaction, water (50 mL) was added to quench and adjusted to acidic with 2M hydrochloric acid, extracted with dichloromethane (50 mL*4), the combined organic phase was washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate and purified by silica gel column chromatography (PE:EA = 5:1) to obtain compound acetyl-L-alanyl-L-valine methyl ester (2.6 g, 10.67 mmol, 58.23% yield), which is ZZL-7.

[0424] LCMS (M+H + ) = 245.3

[0425] 1HNMR (400 MHz, CDC13) δ 6.68 (d, J = 8.5 Hz, 1H), 6.21 (d, J = 7.1 Hz, 1H), 4.57 - 4.47 (m, 1H), 4.44 (dd, J = 8.7, 4.9 Hz, 1H), 3.68 (s, 3H), 2.18 - 2.04 (m, 1H), 1.94 (s, 3H), 1.31 (d, J = 7.0 Hz, 3H), 0.85 (dd, J = 10.1, 6.9 Hz, 6H).

[0426] Preparation of compound A-2-38

[0427]

[0428] First step

[0429] A-2-38a (3.00 g, 13.82 mmol) was dissolved in dichloromethane (50 mL) and stirred for 10 min. Tetrahydro-2H-pyran-4-ol (2.11 g, 20.71 mmol), 4-dimethylaminopyridine (2.52 g, 20.71 mmol) and N,N'-dicyclohexylcarbodiimide (4.27 g, 20.71 mmol) were added at once. After stirring at room temperature for 12 h, TLC showed that the reaction was complete. Water (50 ml) was added to quench, and extracted with dichloromethane (50 ml x 3). The organic phase was combined, washed with saturated brine (200 mL) and dried over anhydrous sodium sulfate. Purification by silica gel column chromatography gave intermediate A-2-38b (2.50 g, 8.30 mmol, 60.0% yield).

[0430] Second step

[0431] Intermediate A-2-38b (2.50 g, 8.30 mmol) was dissolved in dichloromethane (20 ml), and 4M hydrogen chloride solution in dioxane (20 ml) was added. After stirring at room temperature for 2 h, TLC showed that the reaction was complete. Concentration gave the crude intermediate A-2-38c, which was used directly in the next step without purification.

[0432] Third step

[0433] Intermediate A-2-38c crude and intermediate A-2-7b (1.66 g, 8.30 mmol) were dissolved in dichloromethane (40 ml), N,N-diisopropylethylamine (3.22 g, 24.90 mmol), N-hydroxy-7-azabenzotriazole (1.13 g, 8.30 mmol) and l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (1.59 g, 8.30 mmol) were added. After addition, the reaction was stirred in ice bath for 1 h and then was removed to room temperature. After the reaction was stirred overnight, TLC showed the reaction was complete. Water (50 ml) was added and the reaction mixture was extracted with dichloromethane (50 ml x 3). The organic phase was combined, washed with saturated brine (100 ml) and dried over anhydrous sodium sulfate. The product was purified by column chromatography on silica gel to give intermediate A-2-38d (2.00 g, 5.20 mmol, 62.7% overall yield for two steps).

[0434] 1 H NMR (400 MHz, CDC13) δ 5.05 (dt, J = 12.4, 4.1 Hz, 1H), 4.70 (t, J = 8.0 Hz, 1H), 4.57 (dd, J = 8.7, 4.6 Hz, 1H), 4.03 - 3.80 (m, 4H), 3.64 - 3.52 (m, 2H), 2.46 (d, J = 29.1 Hz, 2H), 2.27 (qd, J = 11.7, 6.9 Hz, 1H), 2.03 - 1.90 (m, 2H), 1.73 (ddd, J = 13.1, 8.6, 4.0 Hz, 2H), 1.49 (s, 9H), 0.99 (dd, J = 24.2, 6.9 Hz, 6H).

[0435] Fourth Step

[0436] Intermediate A-2-38d (1.10 g, 2.86 mmol) was dissolved in dichloromethane (5 ml), and 4M hydrogen chloride in dioxane (10 ml) was added. After stirring at room temperature for 2 h, TLC showed the reaction was complete. Concentration gave intermediate A-2-38e crude, which was used directly in the next step without purification.

[0437] Fifth Step

[0438] Intermediate A-2-38e crude was dissolved in tetrahydrofuran (20 ml), and N,N- diisopropylethylamine (1.11 g, 8.58 mmol) was added. After stirring in ice bath for 10 min, acetic anhydride (0.58 g, 5.72 mmol) was added dropwise. After 30 min, the reaction was removed to room temperature. After the reaction was stirred for 2 h, TLC showed the reaction was complete. Ethanol (10 mL) was added, and the reaction mixture was concentrated and purified by column chromatography on silica gel to give product A-2-38 (0.60 g, 1.84 mmol, 64.3% overall yield for two steps).

[0439] 1 H NMR (400 MHz, CDC13) δ 8.28 (d, J = 8.3 Hz, 1H), 4.95 (tt, J = 8.1, 4.0 Hz, 1H), 4.86 (dd, J = 9.5, 6.5 Hz, 1H), 4.41 (dd, J = 8.5, 4.7 Hz, 1H), 4.00 (t, J = 7.7 Hz, 2H), 3.88 - 3.74 (m, 2H), 3.49 (ddd, J = 11.6, 8.3, 3.2 Hz, 2H), 2.61 (ddt, J = 12.0, 8.0, 7.2 Hz, 1H), 2.37 (ddt, J = 12.0, 9.5, 7.2 Hz, 1H), 2.17 (dtd, J = 13.8, 6.9, 4.9 Hz, 1H), 1.88 (s, 3H), 1.87 - 1.81 (m, 2H), 1.69 - 1.53 (m, 2H), 0.90 (dd, J = 19.1, 6.9 Hz, 6H).

[0440] Preparation of compound A-2-39

[0441]

[0442] First step

[0443] A-2-38a (2.00 g, 9.21 mmol) was dissolved in dichloromethane (50 ml), and 2-hydroxyacetic acid methyl ester (1.24 g, 13.81 mmol), 4-dimethylaminopyridine (1.69 g, 13.81 mmol) and N,N'-dicyclohexylcarbodiimide (4.27 g, 20.71 mmol) were added successively under ice-bath. After addition, it was stirred at room temperature for 12 h, TLC showed that the reaction was complete. Water (50 ml) was added, and extracted with dichloromethane (50 ml x 3). The organic phase was combined, washed with saturated brine (200 ml) successively, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to give intermediate A-2-39a (1.50 g, 5.18 mmol, yield 56.2%).

[0444] Second step

[0445] Intermediate A-2-39a (1.50 g, 5.18 mmol) was dissolved in dichloromethane (20 ml), and 4M hydrogen chloride solution in dioxane (20 ml) was added. After stirring at room temperature for 2 h, TLC showed that the reaction was complete. Concentration gave the crude intermediate A-2-39b, which was used directly in the next step without purification.

[0446] Third step

[0447] Intermediate A-2-39b (1.06 g, 5.29 mmol) and intermediate A-2-7b (1.06 g, 5.29 mmol) were dissolved in dichloromethane (40 mL). N,N-diisopropylethylamine (1.92 g, 14.91 mmol), N-hydroxy-7-azabenzotriazole (0.71 g, 5.29 mmol) and l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (1.01 g, 5.29 mmol) were added successively under ice-bath. After addition, the reaction was continued for 1 h under ice-bath and then was allowed to reach room temperature. After overnight, TLC showed the reaction was completed. Water (50 mL) was added and the mixture was extracted with dichloromethane (50 mL x 3). The organic phase was combined and washed successively with saturated brine (100 mL), dried over anhydrous sodium sulfate and purified by silica gel column chromatography to give intermediate A-2-39c (1.10 g, 2.95 mmol, 55.8% over two steps).

[0448] 1 H NMR (400 MHz, CDC13) δ 4.73 (d, J = 15.9 Hz, 1H), 4.60 (t, J = 7.7 Hz, 2H), 4.51 (d, J = 15.9 Hz, 1H), 3.87 (d, J = 8.2 Hz, 1H), 3.77 (dd, J = 14.1, 8.4 Hz, 1H), 3.70 (s, 3H), 2.38 (d, J = 15.2 Hz, 1H), 2.35 - 2.19 (m, 2H), 1.39 (s, 9H), 0.97 (d, J = 6.9 Hz, 3H), 0.93 (d, J = 6.9 Hz, 3H).

[0449] Fourth Step

[0450] Intermediate A-2-39c (1.10 g, 2.95 mmol) was dissolved in dichloromethane (5 mL) and 4 M hydrogen chloride in dioxane (10 mL) was added. After stirring at room temperature for 2 h, TLC showed the reaction was completed. Concentration gave intermediate A-2-39d as a crude product which was used directly in the next step without purification.

[0451] Fifth Step

[0452] Intermediate A-2-39d as a crude product was dissolved in tetrahydrofuran (20 mL) and N,N-diisopropylethylamine (1.14 g, 8.85 mmol) was added. After stirring under ice-bath for 10 min, acetic anhydride (0.60 g, 5.90 mmol) was added dropwise slowly. The reaction was continued for 30 min. After the reaction was completed as shown by TLC, the mixture was allowed to reach room temperature and was continued for 2 h. Ethanol (10 mL) was added and the mixture was concentrated and purified by silica gel column chromatography to give product A-2-39 (0.24 g, 0.76 mmol, 25.8% over two steps).

[0453] 1 H NMR (400 MHz, CDC13) δ 8.30 (d, J = 8.0 Hz, 1H), 4.85 (dd, J = 9.5, 6.6 Hz, 1H), 4.73 (d, J = 15.9 Hz, 1H), 4.55 - 4.50 (m, 1H), 4.50 - 4.44 (m, 1H), 4.08 - 3.93 (m, 2H), 3.69 (s, 3H), 2.60 (ddt, J = 12.0, 9.0, 6.8 Hz, 1H), 2.37 (dtd, J = 11.9, 8.9, 6.0 Hz, 1H), 2.31 - 2.21 (m, 1H), 1.87 (s, 3H), 0.96 (dd, J = 12.9, 6.9 Hz, 6H).

[0454] Preparation of compound A-2-40

[0455]

[0456] First step

[0457] A-2-38a (5.00 g, 23.04 mmol) was dissolved in dichloromethane (50 ml), and 1-methylpiperidin-4-ol (3.97 g, 34.56 mmol), 4-dimethylaminopyridine (4.21 g, 34.56 mmol) and N,N'-dicyclohexylcarbodiimide (7.12 g, 34.56 mmol) were added successively under ice-bath. After addition, it was moved to room temperature and the reaction was continued for 12 h. TLC showed that the reaction was complete. Water (50 ml) was added and extracted with dichloromethane (50 ml x 3). The organic phase was combined and washed with saturated brine (200 ml), dried over anhydrous sodium sulfate and purified by silica gel column chromatography to give intermediate A-2-40a (4.00 g, 12.72 mmol, yield 55.2%).

[0458] Second step

[0459] Intermediate A-2-40a (4.00 g, 12.72 mmol) was dissolved in dichloromethane (20 ml), and 4M hydrogen chloride solution in dioxane (20 ml) was added. After reaction at room temperature for 2 h, TLC showed that the reaction was complete. The crude intermediate A-2-40b was concentrated and used directly in the next step without purification.

[0460] Third step

[0461] Intermediate A-2-40b crude and A-2-7b (2.67 g, 13.36 mmol) were dissolved in dichloromethane (40 ml), N, N-diisopropylethylamine (3.16 g, 24.48 mmol), N-hydroxy-7-azabenzotriazole (1.8 g, 13.22 mmol) and l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (2.55 g, 13.36 mmol) were added under ice-bath. After addition, the reaction was stirred for 1 h at room temperature. After overnight, TLC showed the reaction was complete. Water (50 ml) was added and the mixture was extracted with dichloromethane (50 ml x 3). The organic phase was combined and washed with saturated brine (100 ml), dried over anhydrous sodium sulfate and purified by silica gel column chromatography to give intermediate A-2-40c (2.50 g, 6.29 mmol, 47.1% overall yield for two steps). 1 H NMR (400 MHz, CDC13) δ 4.88 - 4.74 (m, 1H), 4.66 - 4.53 (m, 1H), 4.47 (dd, J = 8.8, 4.6 Hz, 1H), 3.87 (q, J = 8.3 Hz, 1H), 3.76 (td, J = 8.5, 5.4 Hz, 1H), 2.54 (s, 2H), 2.47 - 2.27 (m, 2H), 2.22 (s, 3H), 2.17 (ddd, J = 18.4, 9.2, 4.5 Hz, 2H), 2.04 (s, 1H), 1.91 - 1.78 (m, 2H), 1.68 (dqd, J = 16.6, 8.3, 3.7 Hz, 2H), 1.40 (s, 9H), 0.89 (dd, J = 24.2, 6.9 Hz, 6H).

[0462] Fourth Step

[0463] Intermediate A-2-40c (2.50 g, 6.29 mmol) was dissolved in dichloromethane (5 ml), 4M hydrogen chloride in dioxane (10 ml) was added. After stirring at room temperature for 2 h, TLC showed the reaction was complete. Concentration gave intermediate A-2-40d crude which was used in the next step without purification.

[0464] Fifth Step

[0465] The crude intermediate A-2-40d was dissolved in tetrahydrofuran (20 ml), and N,N-diisopropylethylamine (2.44 g, 18.87 mmol) was added. After stirring in an ice bath for 10 min, acetic anhydride (1.28 g, 12.58 mmol) was added dropwise slowly, and the reaction was continued for 30 min before being moved to room temperature. After the reaction was continued for 2 h, TLC showed that the reaction was complete. Ethanol (10 mL) was added, and the mixture was concentrated and purified by silica gel column chromatography to give the product A-2-40 (1.00 g, 2.95 mmol, 46.8% overall yield for two steps). MS-ESI [M+H] calculated for C17H25N3O4: 340.2; found: 340.3. + 1 H NMR (400 MHz, CDC13) δ 8.35 (d, J = 7.4 Hz, 1H), 5.06 (s, 1H), 4.84 (dd, J = 9.3, 6.6 Hz, 1H), 4.29 (dd, J = 7.5, 5.5 Hz, 1H), 4.02 (t, J = 7.9 Hz, 2H), 3.12 (s, 4H), 2.73 (s, 3H), 2.64 - 2.54 (m, 1H), 2.45 - 2.31 (m, 2H), 2.11 (dt, J = 13.5, 7.0 Hz, 4H), 1.88 (s, 3H), 0.92 (t, J = 7.0 Hz, 6H).

[0466] Preparation of compound A-2-41

[0467]

[0468] First step

[0469] A-2-41a (3.00 g, 13.82 mmol) was dissolved in dichloromethane (50 ml), and 2-methoxyethan-1-ol (1.57 g, 20.73 mmol), 4-dimethylaminopyridine (2.52 g, 20.73 mmol) and N,N'-dicyclohexylcarbodiimide (4.27 g, 20.73 mmol) were added in turn under an ice bath. After addition, the mixture was moved to room temperature, and the reaction was continued for 12 h before TLC showed that the reaction was complete. Water (50 ml) was added, and the mixture was extracted with dichloromethane (50 ml x 3). The combined organic phase was washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to give the intermediate A-2-41a (1.50 g, 5.45 mmol, yield 39.4%).

[0470] Second step

[0471] ​Intermediate A-2-41a (1.50 g, 5.45 mmol) was dissolved in dichloromethane (20 ml), and 4M hydrogen chloride in dioxane (20 ml) was added. After 2 h of reaction at room temperature, TLC showed that the reaction was complete. The crude intermediate A-2-41b was concentrated and used directly in the next step without purification.

[0472] Third Step

[0473] The crude intermediate A-2-41b and A-2-7b (1.21 g, 6.91 mmol) were dissolved in dichloromethane (40 ml), and N,N-diisopropylethylamine (2.11 g, 16.35 mmol), N-hydroxy-7-azabenzotriazole (1.1 g, 8.18 mmol), and 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (1.57 g, 8.18 mmol) were added under ice bath. After 1 h of reaction, the reaction was allowed to reach room temperature. After overnight reaction, TLC showed that the reaction was complete, water (50 ml) was added, and dichloromethane (50 ml x 3) was used for extraction. The organic phase was combined and washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to give intermediate A-2-41c (1.00 g, 2.79 mmol, 51.2% overall yield for two steps). 1 H NMR (400 MHz, CDC13) δ 4.72 - 4.64 (m, 1H), 4.60 (dd, J = 8.9, 4.6 Hz, 1H), 4.40 - 4.32 (m, 1H), 4.31 - 4.23 (m, 1H), 3.94 (q, J = 8.2 Hz, 1H), 3.84 (td, J = 8.5, 5.4 Hz, 1H), 3.64 - 3.55 (m, 2H), 3.38 (s, 3H), 2.42 (d, J = 39.7 Hz, 2H), 2.25 (dtd, J = 13.7, 6.9, 4.9 Hz, 1H), 1.47 (s, 9H), 0.99 (d, J = 6.9 Hz, 3H), 0.93 (d, J = 6.9 Hz, 3H).

[0474] Fourth Step

[0475] Intermediate A-2-41c (1.00 g, 2.79 mmol) was dissolved in dichloromethane (5 ml), and 4M hydrogen chloride in dioxane (10 ml) was added. After 2 h of stirring at room temperature, TLC showed that the reaction was complete. The crude intermediate A-2-41d was concentrated and used directly in the next step without purification.

[0476] Fifth Step

[0477] The crude intermediate A-2-41d was dissolved in tetrahydrofuran (20 ml), and N, N-diisopropyl ethylamine (1.08 g, 8.37 mmol) was added. After stirring for 10 min in an ice bath, acetic anhydride (0.57 g, 5.58 mmol) was added dropwise slowly, and the reaction was continued for 30 min before being brought to room temperature. After the reaction was continued for 2 h, TLC showed that the reaction was complete. Ethanol (10 mL) was added, and the mixture was concentrated and purified by silica gel column chromatography to give the product A-2-41 (0.30 g, 1.00 mmol, 35.8% overall yield for two steps). MS-ESI [M+H] Calc. for C19H25N5O4: 401.2; Found: 401.4. + 1 H NMR (400 MHz, CDC13) δ 8.29 (d, J = 8.1 Hz, 1H), 4.91 (dd, J = 9.4, 6.6 Hz, 1H), 4.52 (dd, J = 8.4, 4.7 Hz, 1H), 4.40 - 4.29 (m, 1H), 4.27 - 4.18 (m, 1H), 4.11 - 4.02 (m, 2H), 3.65 - 3.55 (m, 2H), 3.37 (s, 3H), 2.74 - 2.59 (m, 1H), 2.44 (dtd, J = 11.9, 8.8, 6.3 Hz, 1H), 2.34 - 2.18 (m, 1H), 1.95 (s, 3H), 0.99 (d, J = 6.9 Hz, 3H), 0.95 (d, J = 6.9 Hz, 3H).

[0478] Preparation of compound A-2-42

[0479]

[0480] First step

[0481] A-2-38a (3.00 g, 13.82 mmol) was dissolved in dichloromethane (50 ml), and n-butanol (1.54 g, 20.73 mmol), 4-dimethylaminopyridine (2.52 g, 20.73 mmol) and N, N'-dicyclohexylcarbodiimide (4.27 g, 20.73 mmol) were added in turn under an ice bath. After addition, the mixture was brought to room temperature and the reaction was continued for 12 h, after which TLC showed that the reaction was complete. Water (50 ml) was added, and the mixture was extracted with dichloromethane (50 ml x 3). The combined organic phases were washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to give intermediate A-2-42a (2.50 g, 9.15 mmol, 66.2% yield).

[0482] Second step

[0483] ​Intermediate A-2-42a (2.50 g, 9.15 mmol) was dissolved in dichloromethane (20 ml), and 4 M hydrogen chloride in dioxane (20 ml) was added. After 2 h of reaction at room temperature, TLC showed that the reaction was complete. Concentration gave crude intermediate A-2-42b, which was used directly in the next reaction without purification.

[0484] Third Step

[0485] Intermediate A-2-42b (2.00 g, 5.61 mmol) and A-2-7b (2.21 g, 10.98 mmol) were dissolved in dichloromethane (40 ml), and N,N-diisopropylethylamine (3.55 g, 27.45 mmol), N-hydroxy-7-azabenzotriazole (1.48 g, 10.98 mmol), and 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (2.10 g, 10.98 mmol) were added under ice bath. After 1 h of reaction, the reaction was allowed to reach room temperature. After overnight reaction, TLC showed that the reaction was complete, water (50 ml) was added, and dichloromethane (50 ml x 3) was used for extraction. The organic phase was combined and washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to give intermediate A-2-42c (2.00 g, 5.61 mmol, 61.3% overall yield for two steps). 1 H NMR (400 MHz, CDCl3) δ 4.71 - 4.59 (m, 1H), 4.53 (dd, J = 9.0, 4.7 Hz, 1H), 4.18 - 4.08 (m, 2H), 3.92 (dd, J = 16.4, 8.3 Hz, 1H), 3.81 (td, J = 8.5, 5.4 Hz, 1H), 2.40 (d, J = 40.5 Hz, 2H), 2.26 - 2.14 (m, 1H), 1.65 - 1.57 (m, 2H), 1.45 (s, 9H), 1.38 (dd, J = 15.2, 7.4 Hz, 2H), 0.96 (d, J = 6.9 Hz, 3H), 0.94 - 0.89 (m, 6H).

[0486] Fourth Step

[0487] Intermediate A-2-42c (2.00 g, 5.61 mmol) was dissolved in dichloromethane (5 ml), and 4 M hydrogen chloride in dioxane (10 ml) was added. After 2 h of stirring at room temperature, TLC showed that the reaction was complete. Concentration gave crude intermediate A-2-42d, which was used directly in the next reaction without purification.

[0488] Fifth Step

[0489] The crude intermediate A-2-42d was dissolved in tetrahydrofuran (20 ml), and N,N-diisopropylethylamine (2.18 g, 16.83 mmol) was added. After stirring for 10 min in an ice bath, acetic anhydride (1.15 g, 11.22 mmol) was added dropwise slowly, and the reaction was continued for 30 min before being allowed to warm to room temperature. After 2 h of continued reaction, TLC showed that the reaction was complete. Ethanol (10 mL) was added, and the mixture was concentrated and purified by column chromatography on silica gel to give the product A-2-42 (0.60 g, 2.01 mmol, 35.8% overall yield for two steps). MS-ESI [M+H] Calc’d: 299.2; Found: 299.4. + 1 H NMR (400 MHz, CDC13) δ 8.27 (d, J = 8.3 Hz, 1H), 4.92 (dd, J = 9.5, 6.5 Hz, 1H), 4.48 (dd, J = 8.6, 4.7 Hz, 1H), 4.21 - 4.09 (m, 2H), 4.07 (t, J = 6.0 Hz, 2H), 2.73 - 2.59 (m, 1H), 2.51 - 2.38 (m, 1H), 2.31 - 2.16 (m, 1H), 1.95 (s, 3H), 1.68 - 1.58 (m, 2H), 1.40 (dt, J = 15.1, 7.4 Hz, 2H), 0.99 (d, J = 6.9 Hz, 3H), 0.96 - 0.91 (m, 6H).

[0490] Preparation of compound A-2-43 of Preparation Example 48

[0491]

[0492] First step

[0493] A-2-38a (5.00 g, 23.04 mmol) was dissolved in dichloromethane (50 ml), and cyclopentanol (2.97 g, 34.56 mmol), 4-dimethylaminopyridine (4.21 g, 34.56 mmol), and N,N'-dicyclohexylcarbodiimide (7.12 g, 34.56 mmol) were added sequentially under an ice bath. After addition, the mixture was allowed to warm to room temperature, and the reaction was continued for 12 h, after which TLC showed that the reaction was complete. Water (50 ml) was added, and extraction was performed with dichloromethane (50 ml x 3). The combined organic phases were washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, and purified by column chromatography on silica gel to give intermediate A-2-43a (3.50 g, 12.28 mmol, 53.3% yield).

[0494] Second step

[0495] ​Intermediate A-2-43a (3.50 g, 12.28 mmol) was dissolved in dichloromethane (20 ml) and 4M hydrogen chloride in dioxane (ml) was added. After 2h at room temperature, TLC showed the reaction was complete. The crude intermediate A-2-43b was concentrated and used in the next step without purification.

[0496] Third Step

[0497] The crude intermediate A-2-43b and A-2-7b (2.45 g, 12.28 mmol) were dissolved in dichloromethane (40 ml), N,N-diisopropylethylamine (3.16 g, 24.48 mmol), N-hydroxy-7-azabenzotriazole (1.65 g, 12.28 mmol) and l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (2.34 g, 12.28 mmol) were added under ice-bath. After 1h, the reaction was allowed to warm to room temperature. The reaction was stirred overnight, TLC showed the reaction was complete. Water (50 ml) was added and the mixture was extracted with dichloromethane (50 ml x 3). The organic phase was combined and washed with saturated brine (100 ml), dried over anhydrous sodium sulfate and purified on silica gel column to give intermediate A-2-43c (2.00 g, 5.43 mmol, 44.2% overall yield for two steps). 1 H NMR (400 MHz, CDC13) δ 5.16 (ddd, J = 8.1, 5.8, 2.2 Hz, 1H), 4.67 - 4.52 (m, 1H), 4.44 (dd, J = 9.0, 4.6 Hz, 1H), 3.87 (dd, J = 16.4, 8.2 Hz, 1H), 3.76 (td, J = 8.5, 5.4 Hz, 1H), 2.35 (d, J = 43.0 Hz, 2H), 2.14 (qd, J = 6.8, 2.0 Hz, 1H), 1.84 - 1.75 (m, 2H), 1.72 - 1.59 (m, 4H), 1.55 (dd, J = 7.8, 3.3 Hz, 2H), 1.40 (s, 9H), 0.90 (d, J = 6.9 Hz, 3H), 0.84 (d, J = 6.9 Hz, 3H).

[0498] Fourth Step

[0499] Intermediate A-2-43c (2.00 g, 5.43 mmol) was dissolved in dichloromethane (5 ml) and 4M hydrogen chloride in dioxane (10 ml) was added. After 2h at room temperature, TLC showed the reaction was complete. The crude intermediate A-2-43d was concentrated and used in the next step without purification.

[0500] Fifth Step

[0501] The crude intermediate A-2-43d was dissolved in tetrahydrofuran (20 ml), and N, N-diisopropyl ethylamine (2.11 g, 16.29 mmol) was added. After stirring for 10 min in an ice bath, acetic anhydride (1.11 g, 10.86 mmol) was added dropwise slowly, and the reaction was continued for 30 min before being brought to room temperature. After the reaction was continued for 2 h, TLC showed that the reaction was complete. Ethanol (10 mL) was added, and the mixture was concentrated and purified by silica gel column chromatography to give the product A-2-43 (0.80 g, 2.58 mmol, 47.5% overall yield for two steps). 1 H NMR (400 MHz, CDC13) δ 8.24 (d, J = 8.4 Hz, 1H), 5.21 (td, J = 5.7, 2.9 Hz, 1H), 4.92 (dd, J = 9.5, 6.5 Hz, 1H), 4.45 (dd, J = 8.7, 4.6 Hz, 1H), 4.06 (d, J = 7.2 Hz, 2H), 2.73 - 2.60 (m, 1H), 2.50 - 2.38 (m, 1H), 2.31 - 2.15 (m, 1H), 1.95 (s, 3H), 1.83 (ddd, J = 10.9, 7.3, 5.5 Hz, 2H), 1.78 - 1.65 (m, 4H), 1.61 (dd, J = 7.7, 2.5 Hz, 2H), 0.98 (d, J = 6.9 Hz, 3H), 0.92 (dd, J = 9.9, 4.8 Hz, 3H).

[0502] Preparation of compound A-2-44

[0503]

[0504] First step

[0505] A-2-38a (3.00 g, 13.82 mmol) was dissolved in dichloromethane (50 ml), and cyclopropanol (1.20 g, 20.71 mmol), 4-dimethylaminopyridine (2.52 g, 20.71 mmol) and N, N'-dicyclohexylcarbodiimide (4.27 g, 20.71 mmol) were added in turn under an ice bath. After addition, the mixture was brought to room temperature and the reaction was continued for 12 h, after which TLC showed that the reaction was complete. Water (50 ml) was added, and the mixture was extracted with dichloromethane (50 ml x 3). The combined organic phases were washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to give intermediate A-2-44a (2.10 g, 8.17 mmol, 59.1% yield).

[0506] Second step

[0507] Intermediate A-2-44a (2.10 g, 8.17 mmol) was dissolved in dichloromethane (20 ml), and 4 M hydrogen chloride in dioxane (20 ml) was added. After the reaction was stirred at room temperature for 2 h, TLC showed that the reaction was complete. Concentration gave the crude intermediate A-2-44b, which was used directly in the next reaction without purification.

[0508] Third Step

[0509] The crude intermediate A-2-44b and A-2-7b (1.63 g, 8.16 mmol) were dissolved in dichloromethane (40 ml), and N, N-diisopropylethylamine (3.16 g, 24.48 mmol), N-hydroxy-7-azabenzotriazole (1.10 g, 8.16 mmol), and 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (1.56 g, 8.16 mmol) were added under ice bath. After the addition was completed, the reaction was stirred at room temperature for 1 h, and then was allowed to stand overnight. TLC showed that the reaction was complete, water (50 ml) was added, and extraction was performed with dichloromethane (50 ml x 3). The combined organic phase was washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, and purified by column chromatography on silica gel to give intermediate A-2-44c (1.50 g, 4.41 mmol, 54.0% overall yield for two steps).

[0510] 1 H NMR (400 MHz, CDCl3) δ 4.59 (t, J = 8.0 Hz, 1H), 4.44 (dd, J = 8.9, 4.8 Hz, 1H), 4.16 - 4.08 (m, 1H), 3.87 (dd, J = 16.4, 8.1 Hz, 1H), 3.77 (dd, J = 14.1, 8.4 Hz, 1H), 2.35 (d, J = 34.0 Hz, 2H), 2.16 - 2.05 (m, 1H), 1.40 (s, 9H), 0.90 (d, J = 6.8 Hz, 3H), 0.84 (d, J = 6.9 Hz, 3H), 0.70 - 0.61 (m, 4H).

[0511] Fourth Step

[0512] Intermediate A-2-44c (1.50 g, 4.41 mmol) was dissolved in dichloromethane (5 ml), and 4 M hydrogen chloride in dioxane (10 ml) was added. After the reaction was stirred at room temperature for 2 h, TLC showed that the reaction was complete. Concentration gave the crude intermediate A-2-44d, which was used directly in the next reaction without purification.

[0513] Fifth Step

[0514] The crude intermediate A-2-44d was dissolved in tetrahydrofuran (20 ml), and N,N-diisopropylethylamine (1.71 g, 13.23 mmol) was added. After stirring in an ice bath for 10 min, acetic anhydride (0.90 g, 8.82 mmol) was added dropwise slowly, and the reaction was continued for 30 min before being brought to room temperature. After the reaction was continued for 2 h, TLC showed that the reaction was complete. Ethanol (10 mL) was added, and the mixture was concentrated and purified by column chromatography on silica gel to give the product A-2-44 (0.60 g, 2.12 mmol, 48.2% overall yield for two steps). MS-ESI [M+H] calc’d for C15H22N2O4: 283.2; found: 283.2. + : 283.2; found: 283.2. 1 H NMR (400 MHz, CDC13) δ 8.21 (d, J = 8.3 Hz, 1H), 4.84 (dd, J = 9.5, 6.5 Hz, 1H), 4.36 (dd, J = 8.5, 4.8 Hz, 1H), 4.16 - 4.06 (m, 1H), 4.04 - 3.92 (m, 2H), 2.60 (ddt, J = 12.0, 8.8, 6.9 Hz, 1H), 2.37 (tdd, J = 12.0, 8.8, 6.2 Hz, 1H), 2.20 - 2.06 (m, 1H), 1.88 (s, 3H), 0.90 (d, J = 6.9 Hz, 3H), 0.86 (d, J = 6.9 Hz, 3H), 0.73 - 0.58 (m, 4H).

[0515] Preparation of compounds B-2-12 and B-2-13

[0516]

[0517] First step

[0518] B-2-12a (4.00 g, 37.69 mmol) was dissolved in dichloromethane (50 mL), and (R)-tert-butylsulfinamide (4.57 g, 37.69 mmol) and cesium carbonate (12.28 g, 37.69 mmol) were added in turn. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, and the mixture was filtered, the obtained filtrate was washed with saturated brine (100 x 2 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography on silica gel to give the intermediate B-2-12b (7.00 g, 33.4 mmol, 88.7% yield). MS-ESI [M+H] calc’d for C15H22N2O4: 283.2; found: 283.2. + : 283.2; found: 283.2.

[0519] Second step

[0520] Intermediate B-2-12b (4.50 g, 21.50 mmol) was dissolved in tetrahydrofuran (50 mL). After nitrogen protection and cooling to -78 °C, 1 N isopropyl magnesium bromide tetrahydrofuran solution (40.00 mL, 40.00 mmol) was added dropwise slowly. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was completed. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed with saturated brine (100 mL x 2) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give two components.

[0521] The shorter retention time component 1 was intermediate B-2-12c (1.00 g, 3.94 mmol, yield 18.4%). MS-ESI [M+H] calculated for C15H22NO2 + : 254.2; found: 254.1. 1 H NMR (400 MHz, Chloroform-d) δ 7.29 - 7.23 (m, 2H), 7.22 - 7.14 (m, 3H), 3.38 (d, J = 5.3 Hz, 1H), 2.35 - 1.95 (m, 1H), 1.18 (s, 9H), 0.80 (dd, J = 49.9, 6.8 Hz, 6H).

[0522] The longer retention time component 2 was intermediate B-2-13c (0.24 g, 0.95 mmol, yield: 4.4%). MS-ESI [M+H] calculated for C15H22NO2 + : 254.2; found: 254.1. 1 H NMR (400 MHz, Chloroform-d) δ 7.28 - 7.22 (m, 2H), 7.22 - 7.15 (m, 3H), 4.09 (dd, J = 6.7, 2.2 Hz, 1H), 3.42 (s, 1H), 1.90 (h, J = 6.8 Hz, 1H), 1.12 (s, 9H), 0.83 (dd, J = 72.1, 6.8 Hz, 6H).

[0523] Third step - component 1

[0524] Intermediate B-2-12c (1.00 g, 3.95 mmol) was dissolved in dichloromethane (10 mL), 4 M hydrogen chloride dioxane solution (20 mL) was added under ice bath. After the reaction was continued at ice bath for 1 h, it was concentrated to give intermediate B-2-12d crude product, which was used directly in the next step reaction without purification.

[0525] Fourth step - component 1

[0526] The crude B-2-12d was dissolved in dichloromethane (10 mL) and intermediate A-2-7b (1.08 g, 5.38 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.41 g, 7.33 mmol), 1-hydroxybenzotriazole (0.99 g, 7.33 mmol) and ethyldiisopropylamine (1.90 g, 14.67 mmol) were added successively at -10 °C. After the addition was completed, the reaction was continued at -10 °C for 2 h, then water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave intermediate B-2-12e (0.55 g, 1.65 mmol, 33.8% yield). MS-ESI [M-Boc+2H] Theoretical: 233.2; Found: 233.2. + 1 H NMR (400 MHz, Chloroform-d) δ 7.24 (dd, J = 8.1, 6.4 Hz, 2H), 7.17 (td, J = 7.5, 6.7, 2.0 Hz, 3H), 4.74 (dd, J = 9.4, 7.0 Hz, 1H), 4.66 (s, 1H), 4.05 (q, J = 7.1 Hz, 1H), 3.76 (dq, J = 55.3, 8.2 Hz, 2H), 2.32 (s, 2H), 1.97 - 1.92 (m, 1H), 1.42 (s, 9H), 0.83 (dd, J = 27.0, 6.7 Hz, 6H).

[0527] Fifth Step - Component 1

[0528] Intermediate B-2-12e (0.55 g, 1.65 mmol) was dissolved in dichloromethane (5 mL) and 4 M hydrogen chloride in dioxane (10 mL) was added at ice bath and the reaction was continued for 2 h. The mixture was concentrated to give crude intermediate B-2-12f which was used directly in the next step without purification.

[0529] Sixth Step - Component 1

[0530] The crude intermediate B-2-12f was dissolved in dichloromethane (5 mL) and cooled to -10 °C. Acetyl chloride (0.16 g, 2.08 mmol) and ethyldiisopropylamine (0.73 g, 5.67 mmol) were added. After the reaction was continued at -10 °C for 1 h, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed successively with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave product B-2-12 (86.2 mg, 0.31 mmol, 19.0% yield over two steps). MS-ESI [M+H] Theoretical: 233.2; Found: 233.2.​+ : 275.2; Found: 275.1. 1 H NMR (400 MHz, Chloroform-d) δ 8.51 (d, J = 9.1 Hz, 1H), 7.28 - 7.21 (m, 2H), 7.18 - 7.06 (m, 3H), 4.90 (dd, J = 9.5, 6.5 Hz, 1H), 4.71 (dd, J = 9.1, 6.9 Hz, 1H), 4.02 - 3.81 (m, 2H), 2.59 (ddt, J = 12.0, 9.4, 6.7 Hz, 1H), 2.42 - 2.18 (m, 1H), 1.94 (dt, J = 13.6, 6.7 Hz, 1H), 1.86 (s, 3H), 0.83 (dd, J = 33.1, 6.8 Hz, 6H).

[0531] Third Step - Component 2

[0532] Intermediate B-2-13c (0.24 g, 0.95 mmol) was dissolved in dichloromethane (2 mL), 4M hydrogen chloride solution in dioxane (4 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give crude intermediate B-2-13d which was used directly in the next step without purification.

[0533] Fourth Step - Component 2

[0534] Crude B-2-13d was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (0.27 g, 1.33 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.35 g, 1.81 mmol), 1-hydroxybenzotriazole (0.25 g, 1.81 mmol) and ethyldiisopropylamine (0.47 g, 3.63 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, water (10 mL) was added, and dichloromethane (10 mL x 3) was used for extraction. The organic phase was combined, washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-13e (0.32 g, 0.96 mmol, 100% total yield for two steps). MS-ESI [M-Boc+2H] + : 233.2; Found: 233.2.

[0535] Fifth Step - Component 2

[0536] Intermediate B-2-13e (0.32 g, 0.96 mmol) was dissolved in dichloromethane (3 mL), 4M hydrogen chloride in dioxane (6 mL) was added under ice-bath and the reaction was continued for 2 h, concentrated to give crude intermediate B-2-13f which was used in the next step without purification.

[0537] Step 6 - Component 2

[0538] The crude intermediate B-2-13f was dissolved in dichloromethane (5 mL), cooled to -10 °C, acetyl chloride (97.0 mg, 1.23 mmol) and ethyl diisopropylamine (0.43 g, 3.36 mmol) were added. After 1 h at -10 °C, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave the product B-2-13 (118.2 mg, 0.43 mmol, 44.8% overall yield for two steps). MS-ESI Calcd for [M+H] + : 275.2; Found: 275.1. 1 HNMR (400 MHz, Chloroform-d) δ 8.45 (d, J = 8.8 Hz, 1H), 7.23 (d, J = 7.2 Hz, 2H), 7.20 - 7.12 (m, 3H), 4.79 (dd, J = 9.5, 6.5 Hz, 1H), 4.66 (dd, J = 8.8, 7.0 Hz, 1H), 4.01 (ddd, J = 9.0, 6.5, 3.8 Hz, 2H), 2.78 - 2.49 (m, 1H), 2.42 - 2.22 (m, 1H), 1.95 (dd, J = 13.4, 6.6 Hz, 1H), 1.89 (s, 3H), 0.79 (dd, J = 25.4, 6.7 Hz, 6H).

[0539] Preparation of compounds B-2-14 and B-2-15

[0540]

[0541] First step

[0542] Dissolve B-2-14a (4.00 g, 28.46 mmol) in dichloromethane (50 mL), add (R)-tert-butylsulfinamide (3.45 g, 28.46 mmol) and cesium carbonate (13.91 g, 42.69 mmol) sequentially. After addition, the reaction is allowed to proceed overnight. TLC shows the reaction is complete. Filter, dry the resulting filtrate over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography to give intermediate B-2-14b (6.00 g, 24.6 mmol, 86.5% yield).

[0543] Second Step

[0544] Dissolve intermediate B-2-14b (5.00 g, 20.51 mmol) in tetrahydrofuran (50 mL). After nitrogen protection and cooling to -78 °C, slowly add IN isopropylmagnesium bromide in tetrahydrofuran (41.0 mL, 41.00 mmol) dropwise. After addition, continue the reaction at -78 °C for 2 h. TLC shows the reaction is complete. Adjust the pH to 6 with saturated ammonium chloride solution, and extract with ethyl acetate (100 mL x 3). Combine the organic phases, wash sequentially with saturated brine (100 mL x 2), dry over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography to give two fractions.

[0545] The fraction with shorter retention time is intermediate B-2-14c (0.70 g, 2.43 mmol, 11.9% yield).

[0546] The fraction with longer retention time is intermediate B-2-15c (2.00 g, 6.94 mmol, 33.9% yield).

[0547] Third Step - Fraction 1

[0548] Dissolve intermediate B-2-14c (0.70 g, 2.43 mmol) in dichloromethane (10 mL), and add 4M hydrogen chloride in dioxane (20 mL) under ice bath. Continue the reaction at ice bath for 1 h, concentrate to give crude intermediate B-2-14d, which is used directly in the next step without purification.

[0549] Fourth Step - Fraction 1

[0550] The crude B-2-14d was dissolved in dichloromethane (10 mL) and intermediate A-2-7b (0.65 g, 3.32 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.85 g, 4.41 mmol), 1-hydroxybenzotriazole (0.60 g, 4.41 mmol) and ethyldiisopropylamine (1.14 g, 8.82 mmol) were added successively at -10 °C. After the addition was completed, the reaction was continued at -10 °C for 2 h, then water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified by column chromatography on silica gel to give intermediate B-2-14e (0.45 g, 1.23 mmol, 50.5% overall yield for two steps). MS-ESI [M-Boc+2H] calculated for C26H34N4O6: 267.1; found: 267.1. + : 267.1; found: 267.1.

[0551] Fifth step - Component 1

[0552] Intermediate B-2-14e (0.45 g, 1.23 mmol) was dissolved in dichloromethane (5 mL) and 4 M hydrogen chloride in dioxane (10 mL) was added under ice bath and the reaction was continued for 2 h. The mixture was concentrated to give crude intermediate B-2-14f which was used directly in the next step without purification.

[0553] Sixth step - Component 1

[0554] The crude intermediate B-2-14f was dissolved in dichloromethane (5 mL) and cooled to -10 °C. Acetyl chloride (0.12 g, 1.53 mmol) and ethyldiisopropylamine (0.54 g, 4.17 mmol) were added. After the reaction was continued at -10 °C for 1 h, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed successively with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified by column chromatography on silica gel to give product B-2-14 (155.0 mg, 0.50 mmol, 40.8% overall yield for two steps). MS-ESI [M+H] calculated for C26H34N4O6: 509.2; found: 509.2. + : 267.1; found: 267.1. 1H NMR (400 MHz, Chloroform-d) δ 8.53 (d, J = 8.9 Hz, 1H), 7.21 (d, J = 2.0 Hz, 1H), 7.19 (s, 1H), 7.09 - 7.03 (m, 2H), 4.89 (dd, J = 9.5, 6.5 Hz, 1H), 4.65 (dd, J = 8.8, 6.9 Hz, 1H), 4.08 - 3.82 (m, 2H), 2.45 (ddtd, J = 116.3, 12.0, 9.3, 6.1 Hz, 2H), 1.95 - 1.88 (m, 1H), 1.86 (s, 3H), 0.82 (dd, J = 35.7, 6.7 Hz, 6H).

[0555] Third Step - Component 2

[0556] Intermediate B-2-15c (2.00 g, 6.95 mmol) was dissolved in dichloromethane (20 mL), and 4M hydrogen chloride in dioxane (40 mL) was added under ice bath. After the reaction was continued for 1 h under ice bath, it was concentrated to give crude intermediate B-2-15d which was used directly in the next step without purification.

[0557] Fourth Step - Component 2

[0558] Crude B-2-15d was dissolved in dichloromethane (20 mL), and intermediate A-2-7b (1.81 g, 8.99 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.35 g, 12.25 mmol), 1-hydroxybenzotriazole (1.66 g, 12.25 mmol) and ethyldiisopropylamine (3.17 g, 24.51 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, and water (10 mL) was added, followed by extraction with dichloromethane (20 mL x 3). The organic phase was combined, washed successively with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-15e (0.70 g, 1.91 mmol, 27.5% overall yield for two steps). MS-ESI [M-Boc+2H] Calc’d for C26H32N4O4: 267.1; Found: 267.1. +

[0559] Fifth Step - Component 2

[0560] Intermediate B-2-15e (0.70 g, 1.91 mmol) was dissolved in dichloromethane (10 mL), and 4M hydrogen chloride in dioxane (20 mL) was added under ice bath and the reaction was continued for 2 h. It was concentrated to give crude intermediate B-2-15f which was used directly in the next step without purification.

[0561] ​Step 6 - Component 2

[0562] The crude intermediate B-2-15f was dissolved in dichloromethane (5 mL), cooled to -10 °C, and acetyl chloride (0.16 g, 2.10 mmol) and ethyldiisopropylamine (0.74 g, 5.73 mmol) were added. After 1 h of reaction at -10 °C, water (5 mL) was added, and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the product B-2-15 (74.0 mg, 0.24 mmol, 12.5% overall yield for two steps). MS-ESI Calcd for [M+H] + : 309.1; Found: 309.1. 1 H NMR (400 MHz, Chloroform-d) δ 8.48 (d, J = 8.5 Hz, 1H), 7.33 - 7.17 (m, 2H), 7.16 - 7.00 (m, 2H), 4.79 (dd, J = 9.5, 6.5 Hz, 1H), 4.60 (dd, J = 8.5, 7.0 Hz, 1H), 4.01 (dd, J = 9.0, 6.5 Hz, 2H), 2.66 (dtd, J = 12.0, 8.7, 6.8 Hz, 1H), 2.31 (ddt, J = 12.3, 5.9, 4.7 Hz, 1H), 1.89 (s, 4H), 0.78 (dd, J = 25.4, 6.7 Hz, 6H).

[0563] Preparation of compounds B-2-16 and B-2-17

[0564]

[0565] First step

[0566] B-2-16a (4.00 g, 33.29 mmol) was dissolved in dichloromethane (50 mL), and (R)-tert-butylsulfmamide (4.03 g, 33.29 mmol) and cesium carbonate (16.27 g, 49.94 mmol) were added successively. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, filtered, the obtained filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-16b (6.50 g, 29.1 mmol, 87.4% yield).

[0567] Second step

[0568] Intermediate B-2-16b (5.00 g, 22.39 mmol) was dissolved in tetrahydrofuran (50 mL). After nitrogen protection and cooling to -78 °C, 1 N isopropyl magnesium bromide in tetrahydrofuran (44.5 mL, 40.50 mmol) was added slowly dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was completed. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed with saturated brine (100 mL x 2) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give two components.

[0569] The shorter retention time component 1 was intermediate B-2-16c (1.86 g, 6.96 mmol, 31.1% yield). MS-ESI [M+H] Theoretical: 268.2; Found: 268.1. + 1 H NMR (400 MHz, Chloroform-d) δ 7.19 - 7.03 (m, 4 H), 4.24 (dd, J = 7.7, 6.2 Hz, 1 H), 3.32 (d, J = 6.2 Hz, 1 H), 2.33 (s, 3 H), 2.15 - 1.91 (m, 1 H), 1.14 (s, 9 H), 0.85 (dd, J = 90.1, 6.7 Hz, 6 H).

[0570] The longer retention time component 2 was intermediate B-2-17c (0.50 g, 1.87 mmol, 8.4% yield). MS-ESI [M+H] Theoretical: 268.2; Found: 268.1. + 1 H NMR (400 MHz, Chloroform-d) δ 7.19 - 7.04 (m, 4 H), 4.33 (dd, J = 7.6, 2.6 Hz, 1 H), 3.62 - 3.27 (m, 1 H), 2.32 (s, 3 H), 2.04 - 1.82 (m, 1 H), 1.09 (s, 9 H), 0.87 (dd, J = 88.1, 6.7 Hz, 6 H).

[0571] Third step - component 1

[0572] Intermediate B-2-16c (1.86 g, 6.96 mmol) was dissolved in dichloromethane (15 mL), and 4 M hydrogen chloride in dioxane (30 mL) was added under ice bath. After the reaction was continued at ice bath for 1 h, it was concentrated to give crude intermediate B-2-16d, which was used directly in the next step without purification.

[0573] Fourth step - component 1

[0574] ​​The crude B-2-16d was dissolved in dichloromethane (20 mL) and intermediate A-2-7b (1.36 g, 6.74 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.76 g, 9.20 mmol), 1-hydroxybenzotriazole (1.24 g, 9.20 mmol) and ethyldiisopropylamine (2.38 g, 18.39 mmol) were added successively at -10 °C. After the addition was completed, the reaction was continued at -10 °C for 2 h, then water (10 mL) was added and extracted with dichloromethane (20 mL x 3). The organic phases were combined and washed successively with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave intermediate B-2-16e (1.30 g, 3.75 mmol, 53.9% overall yield for two steps). MS-ESI [M-Boc+2H] calculated for C32H45N5O7: 247.2; found: 247.3. + : 247.2; found: 247.3.

[0575] Fifth step - Component 1

[0576] Intermediate B-2-16e (1.00 g, 2.89 mmol) was dissolved in dichloromethane (10 mL) and 4 M hydrogen chloride in dioxane (20 mL) was added under ice bath and the reaction was continued for 2 h. The mixture was concentrated to give crude intermediate B-2-16f which was used directly in the next step without purification.

[0577] Sixth step - Component 1

[0578] The crude intermediate B-2-16f was dissolved in dichloromethane (10 mL) and cooled to -10 °C. Acetyl chloride (0.25 g, 3.12 mmol) and ethyldiisopropylamine (1.10 g, 8.52 mmol) were added. After the reaction was continued at -10 °C for 1 h, water (10 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave product B-2-16 (331.5 mg, 1.15 mmol, 39.8% overall yield for two steps). MS-ESI [M+H] calculated for C32H45N5O7: 289.2; found: 289.4. + : 247.2; found: 247.3. 1H NMR (400 MHz, Chloroform-d) δ 8.40 (d, J = 8.9 Hz, 1H), 7.13 (t, J = 7.8 Hz, 1H), 7.05 - 6.90 (m, 3H), 4.79 (dd, J = 9.5, 6.4 Hz, 1H), 4.61 (dd, J = 8.9, 7.1 Hz, 1H), 4.10 - 3.91 (m, 2H), 2.78 - 2.60 (m, 1H), 2.30 (dq, J = 9.0, 3.0 Hz, 1H), 2.26 (s, 3H), 1.99 - 1.90 (m, 1H), 1.89 (s, 3H), 0.79 (dd, J = 27.0, 6.8 Hz, 6H).

[0579] Third Step - Component 2

[0580] Intermediate B-2-17c (0.50 g, 1.87 mmol) was dissolved in dichloromethane (5 mL), 4M hydrogen chloride in dioxane (10 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give crude intermediate B-2-17d which was used directly in the next step without purification.

[0581] Fourth Step - Component 2

[0582] Crude B-2-17d was dissolved in dichloromethane (20 mL), and intermediate A-2-7b (0.62 g, 3.10 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.81 g, 4.23 mmol), 1-hydroxybenzotriazole (0.57 g, 4.23 mmol) and ethyldiisopropylamine (1.09 g, 8.46 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, water (10 mL) was added, and dichloromethane (20 mL x 3) was used to extract. The organic phase was combined, washed successively with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-17e (0.50 g, 1.44 mmol, 77.0% total yield over two steps). MS-ESI [M-Boc+2H] Calc: 247.2; Found: 247.4. +

[0583] Fifth Step - Component 2

[0584] Intermediate B-2-17e (0.50 g, 1.44 mmol) was dissolved in dichloromethane (5 mL), 4M hydrogen chloride in dioxane (10 mL) was added under ice-bath and the reaction was continued for 2 h, it was concentrated to give crude intermediate B-2-17f which was used directly in the next step without purification.

[0585] ​Step 6 - Component 2

[0586] The crude intermediate B-2-17f was dissolved in dichloromethane (5 mL), cooled to -10 °C, and acetyl chloride (0.13 g, 1.61 mmol) and ethyldiisopropylamine (0.57 g, 4.38 mmol) were added. After 1 h of reaction at -10 °C, water (5 mL) was added, and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the product B-2-17 (130.4 mg, 0.45 mmol, 31.4% overall yield for two steps). MS-ESI Calcd for [M+H] + : 289.2; Found: 289.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.41 (d, J = 8.9 Hz, 1H), 7.18 - 6.97 (m, 4H), 4.88 (t, J = 8.3 Hz, 1H), 4.75 (dd, J = 9.5, 6.4 Hz, 1H), 4.11 - 3.87 (m, 2H), 2.66 (ddt, J = 12.0, 9.2, 6.7 Hz, 1H), 2.36 (s, 3H), 2.35 - 2.19 (m, 1H), 1.95 (d, J = 23.8 Hz, 1H), 1.89 (s, 3H), 0.82 (dd, J = 49.2, 6.7 Hz, 6H).

[0587] Preparation of compounds B-2-18 and B-2-19

[0588]

[0589] First step

[0590] B-2-18a (4.00 g, 33.29 mmol) was dissolved in dichloromethane (50 mL), and (R)-tert-butylsulfinamide (4.03 g, 33.29 mmol) and cesium carbonate (16.27 g, 49.94 mmol) were added successively. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, filtered, the obtained filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the intermediate B-2-18b (7.20 g, 32.24 mmol, 96.8% yield).

[0591] Second step

[0592] Intermediate B-2-18b (5.00 g, 22.39 mmol) was dissolved in tetrahydrofuran (50 mL). After nitrogen protection and cooling to -78 °C, 1 N isopropylmagnesium bromide in tetrahydrofuran (44.5 mL, 44.50 mmol) was added slowly dropwise. After addition, the reaction was continued at -78 °C for 2 h, and TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phases were combined and washed with saturated brine (100 mL x 2) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give two components.

[0593] The component 1 with shorter retention time was intermediate B-2-18c (0.95 g, 3.55 mmol, 15.9% yield). MS-ESI [M+H] calculated for C17H20N4O4 + : 268.2; found: 268.2.

[0594] The component 2 with longer retention time was intermediate B-2-19c (0.50 g, 1.87 mmol, 8.4% yield). MS-ESI [M+H] calculated for C17H20N4O4 + : 268.2; found: 268.2.

[0595] Third step - component 1

[0596] Intermediate B-2-18c (0.95 g, 3.55 mmol) was dissolved in dichloromethane (10 mL), and a 4 M solution of hydrogen chloride in dioxane (20 mL) was added under ice bath. After the reaction was continued at ice bath for 1 h, it was concentrated to give intermediate B-2-18d crude, which was used directly in the next step without purification.

[0597] Fourth step - component 1

[0598] The intermediate B-2-18d crude was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (1.22 g, 6.06 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.58 g, 8.27 mmol), 1-hydroxybenzotriazole (1.12 g, 8.27 mmol) and ethyldiisopropylamine (2.14 g, 16.53 mmol) were added successively at -10 °C. After addition, the reaction was continued at -10 °C for 2 h, and water (10 mL) was added, extracted with dichloromethane (10 mL x 3). The organic phases were combined and washed with saturated brine (10 mL x 3) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-18e (1.30 g, 3.75 mmol, total yield of two steps 100%). MS-ESI [M-Boc+2H] +: 247.2; found: 247.4.

[0599] Step 5 - Component 1

[0600] Intermediate B-2-18e (1.30 g, 3.75 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride solution in dioxane (20 mL) was added under ice bath and the reaction was continued for 2 h, concentrated to give crude intermediate B-2-18f which was used in the next step without purification.

[0601] Step 6 - Component 1

[0602] Crude intermediate B-2-18f was dissolved in dichloromethane (10 mL), cooled to -10 °C, acetyl chloride (0.35 g, 4.47 mmol) and ethyl diisopropylamine (1.57 g, 12.18 mmol) were added. After 1 h reaction at -10 °C, water (10 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phase was combined and washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated, purified by silica gel column chromatography to give product B-2-18 (192.1 mg, 0.67 mmol, 17.8% total yield over two steps). MS-ESI Calcd for [M+H] + : 289.2; found: 289.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.40 (d, J = 8.9 Hz, 1H), 7.13 (t, J = 7.8 Hz, 1H), 7.04 - 6.86 (m, 3H), 4.79 (dd, J = 9.5, 6.4 Hz, 1H), 4.61 (dd, J = 8.9, 7.1 Hz, 1H), 4.14 - 3.83 (m, 2H), 2.68 (ddt, J = 12.0, 9.1, 6.7 Hz, 1H), 2.34 - 2.27 (m, 1H), 2.26 (s, 3H), 1.95 (d, J = 19.6 Hz, 1H), 1.89 (s, 3H), 0.79 (dd, J = 27.0, 6.8 Hz, 6H).

[0603] Step 3 - Component 2

[0604] Intermediate B-2-19c (0.50 g, 1.87 mmol) was dissolved in dichloromethane (5 mL), 4M hydrogen chloride solution in dioxane (10 mL) was added under ice bath. After 1 h reaction under ice bath, concentrated to give crude intermediate B-2-19d which was used in the next step without purification.

[0605] Step 4 - Component 2

[0606] The crude B-2-19d was dissolved in dichloromethane (10 mL) and intermediate A-2-7b (0.64 g, 3.17 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.83 g, 4.32 mmol), 1-hydroxybenzotriazole (0.58 g, 4.32 mmol) and ethyldiisopropylamine (1.12 g, 8.64 mmol) were added successively at -10 °C. After the addition was completed, the reaction was continued at -10 °C for 2 h, then water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified by silica gel column chromatography to give intermediate B-2-19e (0.55 g, 1.59 mmol, 84.9% overall yield for two steps). MS-ESI [M-Boc+2H] calculated for C26H34N4O6: 247.2; found: 247.3. + : 247.2; found: 247.3.

[0607] Fifth step - Component 2

[0608] Intermediate B-2-19e (0.55 g, 1.59 mmol) was dissolved in dichloromethane (5 mL) and 4 M hydrogen chloride in dioxane (10 mL) was added under ice bath and the reaction was continued for 2 h. The mixture was concentrated to give crude intermediate B-2-19f which was used directly in the next step without purification.

[0609] Sixth step - Component 2

[0610] The crude intermediate B-2-19f was dissolved in dichloromethane (5 mL) and cooled to -10 °C. Acetyl chloride (0.14 g, 1.78 mmol) and ethyldiisopropylamine (0.63 g, 4.63 mmol) were added. After the reaction was continued at -10 °C for 1 h, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed successively with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified by silica gel column chromatography to give product B-2-19 (99.9 mg, 0.35 mmol, 21.8% overall yield for two steps). MS-ESI [M+H] calculated for C26H34N4O6: 289.2; found: 289.4. + : 247.2; found: 247.3. 1H NMR (400 MHz, Chloroform-d) δ 8.47 (d, J = 9.2 Hz, 1H), 7.12 (t, J = 7.5 Hz, 1H), 7.04 - 6.88 (m, 3H), 4.89 (dd, J = 9.6, 6.5 Hz, 1H), 4.68 (dd, J = 9.2, 6.9 Hz, 1H), 4.06 - 3.70 (m, 2H), 2.67 - 2.49 (m, 1H), 2.38 - 2.28 (m, 1H), 2.26 (s, 3H), 2.06 - 1.88 (m, 1H), 1.86 (s, 3H), 0.83 (dd, J = 33.3, 6.8 Hz, 6H).

[0611] Preparation of compounds B-2-20 and B-2-21

[0612]

[0613] First step

[0614] B-2-20a (4.00 g, 33.29 mmol) was dissolved in dichloromethane (50 mL), (R)-tert-butylsulfinamide (4.03 g, 33.29 mmol) and cesium carbonate (16.27 g, 49.94 mmol) were added successively. After addition, the reaction was carried out overnight, TLC showed that the reaction was complete, filtration was carried out, the obtained filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain intermediate B-2-20b (7.00 g, 30.79 mmol, yield 92.5%).

[0615] Second step

[0616] Intermediate B-2-20b (5.00 g, 22.00 mmol) was dissolved in tetrahydrofuran (50 mL). After protection under nitrogen and cooling to -78 °C, 1 N isopropyl magnesium bromide tetrahydrofuran solution (40.00 mL, 40.00 mmol) was slowly added dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed successively with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain two components.

[0617] The component 1 with shorter retention time was intermediate B-2-20c (1.12 g, 4.12 mmol, yield 18.8%).

[0618] The component 2 with longer retention time was intermediate B-2-21c (1.12 g, 4.12 mmol, yield 18.8%).

[0619] Third Step - Component 1

[0620] Intermediate B-2-20c (1.12 g, 4.12 mmol) was dissolved in dichloromethane (10 mL), and 4M hydrogen chloride in dioxane (20 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give crude intermediate B-2-20d which was used directly in the next step without purification.

[0621] Fourth Step - Component 1

[0622] Crude intermediate B-2-20d was dissolved in dichloromethane (20 mL), and intermediate A-2-7b (1.39 g, 6.91 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.81 g, 9.42 mmol), 1-hydroxybenzotriazole (1.27 g, 9.42 mmol) and ethyldiisopropylamine (2.43 g, 18.84 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, and water (10 mL) was added, followed by extraction with dichloromethane (20 mL x 3). The organic phases were combined, washed successively with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-20e (0.55 g, 1.57 mmol, 38.1% overall yield for two steps). MS-ESI [M-Boc+2H] Theor: 251.2; Found: 251.2. +

[0623] Fifth Step - Component 1

[0624] Intermediate B-2-20e (0.55 g, 1.57 mmol) was dissolved in dichloromethane (5 mL), and 4M hydrogen chloride in dioxane (10 mL) was added under ice-bath and the reaction was continued for 2 h. It was concentrated to give crude intermediate B-2-20f which was used directly in the next step without purification.

[0625] Sixth Step - Component 1

[0626] Crude intermediate B-2-20f was dissolved in dichloromethane (5 mL), and acetyl chloride (0.14 g, 1.80 mmol) and ethyldiisopropylamine (0.64 g, 4.92 mmol) were added under -10 °C. After the reaction was continued for 1 h under -10 °C, water (5 mL) was added, followed by extraction with dichloromethane (5 mL x 3). The organic phases were combined, washed with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give product B-2-20 (76.7 mg, 0.26 mmol, 16.7% overall yield for two steps). MS-ESI [M+H]​+ : 293.2; Found: 293.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.46 (d, J = 8.6 Hz, 1H), 7.19 - 7.11 (m, 2H), 6.93 (t, J = 8.7 Hz, 2H), 4.79 (dd, J = 9.5, 6.5 Hz, 1H), 4.61 (dd, J = 8.6, 7.1 Hz, 1H), 4.06 - 3.88 (m, 2H), 2.76 - 2.49 (m, 1H), 2.31 (dddd, J = 12.0, 9.5, 8.3, 6.3 Hz, 1H), 1.92 (d, J = 7.0 Hz, 1H), 1.89 (s, 3H), 0.78 (dd, J = 29.5, 6.7 Hz, 6H).

[0627] Third Step - Component 2

[0628] Intermediate B-2-21c (1.12 g, 4.19 mmol) was dissolved in dichloromethane (10 mL), and 4M hydrogen chloride in dioxane (20 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give crude intermediate B-2-21d which was used directly in the next step without purification.

[0629] Fourth Step - Component 2

[0630] The crude B-2-21d was dissolved in dichloromethane (20 mL), and intermediate A-2-7b (1.39 g, 6.91 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.81 g, 9.42 mmol), 1-hydroxybenzotriazole (1.27 g, 9.42 mmol) and ethyldiisopropylamine (2.43 g, 18.84 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, and water (10 mL) was added, followed by extraction with dichloromethane (20 mL x 3). The combined organic phase was washed successively with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave intermediate B-2-21e (1.50 g, 4.28 mmol, 100% overall yield for two steps). MS-ESI [M-Boc+2H] + : 251.1; Found: 251.3.

[0631] Fifth Step - Component 2

[0632] Intermediate B-2-21e (1.00 g, 2.85 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride in dioxane (20 mL) was added under ice-bath and the reaction was continued for 2 h, concentrated to give crude intermediate B-2-21f which was used in the next step without purification.

[0633] Step 6 - Component 2

[0634] Intermediate B-2-21f was dissolved in dichloromethane (10 mL), cooled to -10 °C, acetyl chloride (0.28 g, 3.52 mmol) and ethyl diisopropylamine (1.24 g, 9.60 mmol) were added. After 1 h at -10 °C, water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phases were combined and washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification on a silica gel column gave product B-2-21 (0.49 g, 1.68 mmol, 58.8% overall yield for two steps). MS-ESI calc. for [M+H] + : 293.2; found: 293.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.65 - 8.34 (m, 1H), 7.12 - 7.02 (m, 2H), 7.00 - 6.79 (m, 2H), 4.89 (dd, J = 9.5, 6.5 Hz, 1H), 4.67 (dd, J = 8.9, 7.0 Hz, 1H), 4.13 - 3.79 (m, 2H), 2.60 (ddt, J = 12.0, 9.4, 6.7 Hz, 1H), 2.31 (dtd, J = 12.0, 9.3, 5.5 Hz, 1H), 1.95 - 1.87 (m, 1H), 1.86 (s, 3H), 0.82 (dd, J = 38.1, 6.7 Hz, 6H).

[0635] Preparation of compounds B-2-22 and B-2-23

[0636]

[0637] Step 1

[0638] B-2-22a (4.00 g, 22.79 mmol) was dissolved in dichloromethane (50 mL), (R)-tert-butylsulfinamide (2.78 g, 22.97 mmol) and cesium carbonate (11.23 g, 34.45 mmol) were added successively. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, the resulting filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-22b (6.00 g, 21.6 mmol, 94.9% yield).

[0639] Second step

[0640] Intermediate B-2-22b (5.00 g, 18.03 mmol) was dissolved in tetrahydrofuran (50 mL). After nitrogen protection and cooling to -78 °C, 1 N isopropyl magnesium bromide in tetrahydrofuran (36.0 mL, 36.0 mmol) was added slowly dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed successively with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give two components.

[0641] The shorter retention time component 1 was intermediate B-2-22c (1.00 g, 3.11 mmol, 17.3% yield). MS-ESI [M+H] Theoretical value: 322.1; Found: 322.4. + 1 H NMR (400 MHz, Chloroform-d) δ 7.67-7.48 (m, 2H), 7.38-7.28 (m, 2H), 4.12 (t, J = 6.3 Hz, 1H), 3.41 (d, J = 6.4 Hz, 1H), 2.42-1.95 (m, 1H), 1.18 (s, 9H), 0.81 (dd, J = 52.3, 6.7 Hz, 6H).

[0642] The longer retention time component 2 was intermediate B-2-23c (1.00 g, 3.11 mmol, 17.3% yield). MS-ESI [M+H] Theoretical value: 322.1; Found: 322.4. + 1 ​​H NMR (400 MHz, Chloroform-d) δ 7.51 (d, J = 8.1 Hz, 2H), 7.31 (d, J = 8.0 Hz, 2H), 4.17 (dd, J = 6.6, 2.0 Hz, 1H), 3.46 (d, J = 2.0 Hz, 1H), 1.95 - 1.87 (m, 1H), 1.13 (s, 9H), 0.83 (dd, J = 69.5, 6.8 Hz, 6H).

[0643] Third Step - Component 1

[0644] Intermediate B-2-22c (1.00 g, 3.11 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride in dioxane (20 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give crude intermediate B-2-22d which was used directly in the next step without purification.

[0645] Fourth Step - Component 1

[0646] Crude intermediate B-2-22d was dissolved in dichloromethane (20 mL), and intermediate A-2-7b (1.02 g, 5.06 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.32 g, 6.90 mmol), 1-hydroxybenzotriazole (0.93 g, 6.90 mmol) and ethyldiisopropylamine (1.78 g, 13.80 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, water (10 mL) was added, and dichloromethane (20 mL x 3) was used to extract. The organic phase was combined, washed successively with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-22e (1.10 g, 2.75 mmol, 88.3% overall yield for two steps).

[0647] Fifth Step - Component 1

[0648] Intermediate B-2-22e (1.10 g, 2.75 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride in dioxane (20 mL) was added under ice-bath and the reaction was continued for 2 h, it was concentrated to give crude intermediate B-2-22f which was used directly in the next step without purification.

[0649] Sixth Step - Component 1

[0650] The crude intermediate B-2-22f was dissolved in dichloromethane (10 mL), cooled to -10 °C, acetyl chloride (0.23 g, 2.93 mmol) and ethyl diisopropylamine (1.03 g, 7.98 mmol) were added. After 1 h of reaction at -10 °C, water (10 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phase was combined and washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give the product B-2-22 (152.1 mg, 0.44 mmol, 16.2% overall yield for two steps). MS-ESI [M+H] calc’d for C17H22F2N4O2: 343.2; found: 343.2. + 1 H NMR (400 MHz, Chloroform-d) δ 8.58 (d, J = 8.3 Hz, 1H), 7.50 (d, J = 8.1 Hz, 2H), 7.32 (d, J = 8.1 Hz, 2H), 4.80 (dd, J = 9.5, 6.5 Hz, 1H), 4.67 (dd, J = 8.4, 6.9 Hz, 1H), 4.06 - 3.95 (m, 2H), 2.73 - 2.58 (m, 1H), 2.31 (ddt, J = 12.1, 9.5, 6.9 Hz, 1H), 2.00 - 1.92 (m, 1H), 1.90 (s, 3H), 0.80 (dd, J = 22.1, 6.8 Hz, 6H).

[0651] Third Step - Component 2

[0652] The intermediate B-2-23c (1.00 g, 3.11 mmol) was dissolved in dichloromethane (10 mL), 4 M hydrogen chloride solution in dioxane (20 mL) was added under ice bath. After 1 h of reaction under ice bath, it was concentrated to give the crude intermediate B-2-23d which was used in the next step reaction without purification.

[0653] Fourth Step - Component 2

[0654] ​The crude B-2-23d was dissolved in dichloromethane (20 mL) and intermediate A-2-7b (1.07 g, 5.31 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.39 g, 7.25 mmol), 1-hydroxybenzotriazole (0.98 g, 7.25 mmol) and ethyldiisopropylamine (1.87 g, 14.49 mmol) were added successively at -10 °C. After the addition was completed, the reaction was continued at -10 °C for 2 h, then water (10 mL) was added and extracted with dichloromethane (20 mL x 3). The organic phases were combined and washed successively with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified by column chromatography on silica gel to give intermediate B-2-23e (1.00 g, 2.50 mmol, 80.3% overall yield for two steps).

[0655] Fifth step - Component 2

[0656] Intermediate B-2-23e (1.00 g, 2.50 mmol) was dissolved in dichloromethane (10 mL) and 4 M hydrogen chloride in dioxane (20 mL) was added under ice bath and the reaction was continued for 2 h. The mixture was concentrated to give crude intermediate B-2-23f which was used directly in the next step without purification.

[0657] Sixth step - Component 2

[0658] The crude intermediate B-2-23f was dissolved in dichloromethane (10 mL) and cooled to -10 °C. Acetyl chloride (0.20 g, 2.56 mmol) and ethyldiisopropylamine (0.90 g, 6.99 mmol) were added. After the reaction was continued at -10 °C for 1 h, water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified by column chromatography on silica gel to give product B-2-23 (284.6 mg, 0.83 mmol, 33.2% overall yield for two steps). MS-ESI [M+H] calc’d for C26H30N4O5: 343.2; found: 343.2. + 1 ​H NMR (400 MHz, Chloroform-d) δ 8.64 (d, J = 8.8 Hz, 1H), 7.49 (d, J = 8.0 Hz, 2H), 7.24 (d, J = 8.1 Hz, 2H), 4.90 (dd, J = 9.5, 6.5 Hz, 1H), 4.74 (dd, J = 8.8, 6.8 Hz, 1H), 4.08 - 3.78 (m, 2H), 2.68 - 2.52 (m, 1H), 2.31 (dtd, J = 12.0, 9.3, 5.5 Hz, 1H), 2.01 - 1.91 (m, 1H), 1.88 (s, 3H), 0.84 (dd, J = 30.1, 6.8 Hz, 6H).

[0659] Preparation of compound B-2-24

[0660]

[0661] First step

[0662] B-2-24a (4.00 g, 29.38 mmol) was dissolved in dichloromethane (50 mL), (R)-tert-butylsulfinamide (4.57 g, 29.38 mmol) and anhydrous copper sulfate (4.69 g, 29.38 mmol) were added successively. After addition, the reaction was carried overnight, TLC showed that the reaction was complete, filtered, the obtained filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain intermediate B-2-24b (4.00 g, 16.7 mmol, yield 56.9%). MS-ESI [M+H] calculated: 240.1; found: 240.1. +

[0663] Second step

[0664] Intermediate B-2-24b (2.00 g, 8.36 mmol) was dissolved in tetrahydrofuran (20 mL). After nitrogen protection and cooling to -78°C, 1N isopropyl magnesium bromide tetrahydrofuran solution (16.72 mL, 16.72 mmol) was slowly added dropwise. After addition, the reaction was continued at -78°C for 2h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed successively with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain a mixture of two components, recorded as intermediate B-2-24c (1.00 g, 3.53 mmol, yield 42.2%). MS-ESI [M+H] calculated: 284.2; found: 284.1. +

[0665] ​​Step 3

[0666] The intermediate B-2-24c component 1 (1.00 g, 3.53 mmol) was dissolved in dichloromethane (10 mL), and 4M hydrogen chloride in dioxane (20 mL) was added under ice bath. After the reaction was continued for 1 h under ice bath, it was concentrated to give the crude intermediate B-2-24d which was used directly in the next step without purification.

[0667] Step 4

[0668] The crude intermediate B-2-24d was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (1.11 g, 5.02 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.44 g, 7.53 mmol), 1-hydroxybenzotriazole (1.02 g, 7.53 mmol) and ethyldiisopropylamine (1.95 g, 15.06 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, and water (10 mL) was added, followed by extraction with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give intermediate B-2-24e (1.20 g, 3.31 mmol, 93.8% overall yield for two steps). MS-ESI [M-Boc+2H] + : 263.2; found: 263.2.

[0669] Step 5

[0670] The intermediate B-2-24e (1.20 g, 3.31 mmol) was dissolved in dichloromethane (10 mL), and 4M hydrogen chloride in dioxane (20 mL) was added under ice bath and the reaction was continued for 2 h. It was concentrated to give the crude intermediate B-2-24f which was used directly in the next step without purification.

[0671] Step 6 - Component 1

[0672] The crude intermediate B-2-24f was dissolved in dichloromethane (10 mL), and acetyl chloride (0.28 g, 3.61 mmol) and ethyldiisopropylamine (1.27 g, 9.84 mmol) were added under -10 °C. After the reaction was continued for 1 h under -10 °C, water (10 mL) was added, followed by extraction with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give the product B-2-24 (332.9 mg, 1.09 mmol, 33.0% overall yield for two steps). MS-ESI [M+H] + : 305.2; found: 305.2.1 H NMR(400MHz,Chloroform-d)δ8.40(dd,J=25.0,8.9Hz,1H),7.14–7.00(m,2H),6.81–6.72(m,2H),4.83(ddd,J=41.8,9.5,6.5Hz,1H),4.62(ddd,J=21.1,8. 9,7.1Hz,1H),4.16–3.86(m,2H),3.71(d,J=2.8Hz,3H),2.79–2.47(m,1H),2. 43–2.17(m,1H),1.99–1.90(m,1H),1.87(d,J=15.0Hz,3H),0.89–0.64(m,6H).

[0673] Preparation of compound B-2-25 in Example 57

[0674]

[0675] first step

[0676] B-2-25a (5.00 g, 36.72 mmol) was dissolved in dichloromethane (50 mL), and (R)-tert-butylsulfinamide (4.90 g, 40.39 mmol) and anhydrous copper sulfate (5.86 g, 36.72 mmol) were added sequentially. After the addition was complete, the reaction was allowed to proceed overnight. TLC showed that the reaction was complete. The mixture was filtered, and the resulting filtrate was washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-25b (8.00 g, 27.28 mmol, yield 74.3%). MS-ESI theoretical value [M+H] + : 240.1; Measured value: 240.1.

[0677] Step 2

[0678] Intermediate B-2-25b (5.00 g, 17.05 mmol) was dissolved in tetrahydrofuran (50 mL). Under nitrogen protection and cooling to -78 °C, a solution of 1N isopropyl magnesium bromide in tetrahydrofuran (41.78 mL, 41.78 mmol) was slowly added dropwise. After the addition was complete, the reaction was continued at -78 °C for 2 h, and TLC showed completion. The pH was adjusted to 6 with saturated ammonium chloride solution, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and washed successively with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give a mixture of two components, designated as intermediate B-2-25c (2.30 g, 8.12 mmol, yield 47.6%). MS-ESI theoretical value [M+H] +: 284.2; Found: 284.2.

[0679] Third Step

[0680] The intermediate B-2-25c component 1 (1.30 g, 4.59 mmol) was dissolved in dichloromethane (10 mL), and 4M hydrogen chloride solution in dioxane (20 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give the crude intermediate B-2-25d which was used directly in the next step without purification.

[0681] Fourth Step

[0682] The crude B-2-25d was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (1.45 g, 7.36 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.92 g, 10.04 mmol), 1-hydroxybenzotriazole (1.36 g, 10.04 mmol) and ethyldiisopropylamine (2.59 g, 20.07 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, and then water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phase was combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave intermediate B-2-25e (1.50 g, 4.14 mmol, 90.2% overall yield for two steps). MS-ESI Calcd for [M-Boc+2H] + : 263.2; Found: 263.4.

[0683] Fifth Step

[0684] Intermediate B-2-25e (1.50 g, 4.14 mmol) was dissolved in dichloromethane (10 mL), and 4M hydrogen chloride solution in dioxane (20 mL) was added under ice-bath and the reaction was continued for 2 h. It was concentrated to give the crude intermediate B-2-25f which was used directly in the next step without purification.

[0685] Sixth Step

[0686] The crude intermediate B-2-25f was dissolved in dichloromethane (10 mL), cooled to -10 °C, and acetyl chloride (0.33 g, 4.19 mmol) and ethyldiisopropylamine (1.48 g, 11.43 mmol) were added. After 1 h of reaction at -10 °C, water (10 mL) was added, and extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the product B-2-25 (431.5 g, 1.44 mmol, 34.9% overall yield for two steps). MS-ESI [M+H] calc’d for C17H21F2NO3, 305.2; found, 305.2. + : 305.2; found, 305.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.36 (t, J = 12.2 Hz, 1H), 7.17 - 6.99 (m, 2H), 6.86 - 6.78 (m, 2H), 4.91 - 4.81 (m, 1H), 4.77 - 4.57 (m, 1H), 4.05 - 3.90 (m, 2H), 3.78 (d, J = 13.5 Hz, 3H), 2.70 - 2.48 (m, 1H), 2.32 (ddtd, J = 15.1, 11.9, 9.3, 5.7 Hz, 1H), 2.13 - 2.01 (m, 1H), 1.86 (d, J = 18.2 Hz, 3H), 0.90 - 0.69 (m, 6H).

[0687] Preparation of compound B-2-26

[0688]

[0689] First step

[0690] B-2-26a (5.00 g, 36.72 mmol) was dissolved in dichloromethane (50 mL), and (R)-tert-butylsulfinamide (4.90 g, 40.39 mmol) and anhydrous copper sulfate (5.86 g, 36.72 mmol) were added successively. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, filtered, the obtained filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the intermediate B-2-26b (6.00 g, 25.07 mmol, yield 68.3%). MS-ESI [M+H] calc’d for C17H21F2NO3, 305.2; found, 305.2. + : 305.2; found, 305.2.

[0691] Second step

[0692] Intermediate B-2-26b (5.00 g, 20.89 mmol) was dissolved in tetrahydrofuran (50 mL). After nitrogen protection and cooling to -78 °C, 1 N isopropyl magnesium bromide in tetrahydrofuran (41.78 mL, 41.78 mmol) was added dropwise slowly. After addition, the reaction was continued at -78 °C for 2 h, TLC showed the reaction was completed. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed with saturated brine (100 mL x 2) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give a mixture of two components, recorded as intermediate B-2-26c (2.30 g, 8.12 mmol, 38.8% yield). MS-ESI [M+H] Theoretical value: 284.2; Found: 284.1. + : 284.2; Found: 284.1.

[0693] Third Step

[0694] Intermediate B-2-26c (1.50 g, 5.29 mmol) was dissolved in dichloromethane (10 mL), and 4 M hydrogen chloride in dioxane (20 mL) was added under ice bath. After the reaction was continued at ice bath for 1 h, it was concentrated to give intermediate B-2-26d crude product, which was used directly in the next step reaction without purification.

[0695] Fourth Step

[0696] The intermediate B-2-26d crude product was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (1.73 g, 8.59 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (2.25 g, 11.71 mmol), 1-hydroxybenzotriazole (1.58 g, 11.71 mmol) and ethyldiisopropylamine (3.03 g, 23.43 mmol) were added successively under -10 °C. After addition, the reaction was continued at -10 °C for 2 h, and then water (10 mL) was added, extracted with dichloromethane (10 mL x 3). The organic phase was combined and washed with saturated brine (10 mL x 3) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-26e (1.70 g, 4.69 mmol, total yield of two steps 88.7%). MS-ESI [M-Boc+2H] Theoretical value: 263.2; Found: 263.2. + : 263.2; Found: 263.2.

[0697] Fifth Step

[0698] Intermediate B-2-26e (1.00 g, 2.76 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride solution in dioxane (20 mL) was added under ice-bath and the reaction was continued for 2 h, concentrated to give crude intermediate B-2-26f which was used in the next step without purification.

[0699] Step 6

[0700] The crude intermediate B-2-26f was dissolved in dichloromethane (10 mL), cooled to -10 °C, acetyl chloride (0.24 g, 3.01 mmol) and ethyl diisopropylamine (1.06 g, 8.22 mmol) were added. After 1 h at -10 °C, water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phase was combined and washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated, purified by silica gel column chromatography to give product B-2-26 (199.8 mg, 0.66 mmol, 23.8% overall yield for two steps). MS-ESI calc. for [M+H] + : 305.2; found: 305.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.40 (dd, J = 25.0, 8.9 Hz, 1H), 7.14 - 7.00 (m, 2H), 6.81 - 6.72 (m, 2H), 4.83 (ddd, J = 41.8, 9.5, 6.5 Hz, 1H), 4.62 (ddd, J = 21.1, 8.9, 7.1 Hz, 1H), 4.16 - 3.86 (m, 2H), 3.71 (d, J = 2.8 Hz, 3H), 2.79 - 2.47 (m, 1H), 2.43 - 2.17 (m, 1H), 1.99 - 1.90 (m, 1H), 1.87 (d, J = 15.0 Hz, 3H), 0.89 - 0.64 (m, 6H).

[0701] Preparation of compounds B-2-27 and B-2-28

[0702]

[0703] Step 1

[0704] B-2-27a (5.00 g, 34.21 mmol) was dissolved in dichloromethane (50 mL), (R)-tert-butylsulfinamide (4.56 g, 37.63 mmol) and cesium carbonate (22.29 g, 68.42 mmol) were added successively. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, filtration was performed, the obtained filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-27b (4.00 g, 16.04 mmol, 46.9% yield). MS-ESI [M+H] calculated for C15H22NO3S, 250.1; found, 250.1. + : 250.1; found, 250.1.

[0705] Second step

[0706] Intermediate B-2-27b (4.00 g, 16.04 mmol) was dissolved in tetrahydrofuran (50 mL). After being protected by nitrogen and cooled to -78 °C, 1N isopropyl magnesium bromide tetrahydrofuran solution (32.00 mL, 32.00 mmol) was added dropwise slowly. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed successively with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give two components.

[0707] The component 1 with shorter retention time was intermediate B-2-27c (1.30 g, 4.43 mmol, 27.6% yield). MS-ESI [M+H] calculated for C16H24NO3S, 294.1; found, 294.1. + : 250.1; found, 250.1. 1 H NMR (400 MHz, Chloroform-d) δ 7.53-7.28 (m, 2H), 7.18-7.04 (m, 2H), 6.64 (d, J = 0.8 Hz, 1H), 4.18 (ddd, J = 8.8, 6.2, 0.7 Hz, 1H), 3.54 (d, J = 8.7 Hz, 1H), 2.26 (dq, J = 13.4, 6.7 Hz, 1H), 1.19 (d, J = 5.3 Hz, 9H), 0.90 (dd, J = 12.3, 6.8 Hz, 6H).

[0708] The component 2 with longer retention time was intermediate B-2-28c (1.00 g, 3.41 mmol, 21.2% yield). MS-ESI [M+H] calculated for C16H24NO3S, 294.1; found, 294.1. + : 250.1; found, 250.1. 1HNMR (400 MHz, Chloroform-d) δ 7.58 - 7.30 (m, 2H), 7.22 - 7.12 (m, 2H), 6.52 (t, J = 0.7 Hz, 1H), 4.30 (dd, J = 6.4, 4.7 Hz, 1H), 3.46 (d, J = 4.7 Hz, 1H), 2.29 - 2.06 (m, 1H), 1.13 (s, 9H), 0.94 (dd, J = 29.7, 6.8 Hz, 6H).

[0709] Third Step - Component 1

[0710] Intermediate B-2-27c (1.00 g, 3.41 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride in dioxane (10 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give crude intermediate B-2-27d which was used directly in the next step without purification.

[0711] Fourth Step - Component 1

[0712] Crude B-2-27d was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (0.94 g, 4.65 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.22 g, 6.35 mmol), 1-hydroxybenzotriazole (0.86 g, 6.35 mmol) and ethyldiisopropylamine (1.64 g, 12.69 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, water (10 mL) was added, and dichloromethane (10 mL x 3) was used for extraction. The organic phase was combined, washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-27e (0.80 g, 2.15 mmol, 63.0% overall yield for two steps). MS-ESI [M-Boc+2H] + : 273.2; found: 273.2.

[0713] Fifth Step - Component 1

[0714] Intermediate B-2-27e (0.50 g, 1.34 mmol) was dissolved in dichloromethane (5 mL), 4M hydrogen chloride in dioxane (5 mL) was added under ice-bath and the reaction was continued for 2 h, it was concentrated to give crude intermediate B-2-27f which was used directly in the next step without purification.

[0715] Sixth Step - Component 1

[0716] The crude intermediate B-2-27f was dissolved in dichloromethane (5 mL), cooled to -10 °C, and acetyl chloride (0.12 g, 1.50 mmol) and ethyldiisopropylamine (0.53 g, 4.08 mmol) were added. After 1 h of reaction at -10 °C, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phase was combined and washed with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the product B-2-27 (75.0 mg, 0.24 mmol, 17.8% total yield over two steps). MS-ESI calc. for [M+H] + : 315.2; found: 315.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.44 (d, J = 9.0 Hz, 1H), 7.53 - 7.41 (m, 2H), 7.20 (dtd, J = 18.6, 7.3, 1.3 Hz, 2H), 6.59 (d, J = 0.9 Hz, 1H), 4.97 (ddd, J = 53.4, 9.2, 6.4 Hz, 2H), 4.27 - 3.94 (m, 2H), 2.76 (ddt, J = 12.0, 9.0, 6.7 Hz, 1H), 2.45 - 2.35 (m, 1H), 2.34 - 2.22 (m, 1H), 1.96 (s, 3H), 0.94 (dd, J = 6.8, 1.2 Hz, 6H).

[0717] Third Step - Component 2

[0718] The intermediate B-2-28c (1.00 g, 3.41 mmol) was dissolved in dichloromethane (2 mL), and 4 M hydrogen chloride solution in dioxane (4 mL) was added under ice bath. After 1 h of reaction under ice bath, it was concentrated to give the crude intermediate B-2-28d, which was used directly in the next step without purification.

[0719] Fourth Step - Component 2

[0720] The crude B-2-28d was dissolved in dichloromethane (10 mL) and intermediate A-2-7b (0.82 g, 4.07 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.06 g, 5.55 mmol), 1-hydroxybenzotriazole (0.75 g, 5.55 mmol) and ethyldiisopropylamine (1.43 g, 11.10 mmol) were added successively at -10 °C. After the addition, the reaction was continued at -10 °C for 2 h, then water (10 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified by column chromatography on silica gel to give intermediate B-2-28e (0.80 g, 2.15 mmol, 63.0% overall yield for two steps).

[0721] Fifth step - Component 2

[0722] Intermediate B-2-28e (0.80 g, 2.15 mmol) was dissolved in dichloromethane (10 mL) and 4 M hydrogen chloride in dioxane (10 mL) was added under ice bath and the reaction was continued for 2 h. The mixture was concentrated to give crude intermediate B-2-28f which was used directly in the next step without purification.

[0723] Sixth step - Component 2

[0724] The crude intermediate B-2-28f was dissolved in dichloromethane (5 mL) and cooled to -10 °C. Acetyl chloride (0.18 g, 2.34 mmol) and ethyldiisopropylamine (0.83 g, 6.39 mmol) were added. After the reaction was continued at -10 °C for 1 h, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed successively with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product B-2-28 (189.0 g, 0.60 mmol, 28.0% overall yield for two steps) was purified by column chromatography on silica gel. MS - ESI [M+H] calc’d for C19H21F2N3O4: 315.2; found: 315.4. + 1 HNMR (400 MHz, Chloroform-d) δ 8.46 (d, J = 9.1 Hz, 1H), 7.60 - 7.38 (m, 2H), 7.26 - 7.17 (m, 2H), 6.51 (d, J = 0.9 Hz, 1H), 5.24 - 4.87 (m, 2H), 4.12 - 3.82 (m, 2H), 2.68 (ddt, J = 12.0, 9.2, 6.9 Hz, 1H), 2.49 - 2.36 (m, 1H), 2.35 - 2.15 (m, 1H), 1.94 (s, 3H), 0.98 (dd, J = 9.6, 6.8 Hz, 6H). ​

[0725] Preparation of compounds B-2-29 and B-2-30

[0726]

[0727] First step

[0728] B-2-29a (5.00 g, 27.44 mmol) was dissolved in dichloromethane (50 mL), (R)- tert-butylsulfinamide (3.66 g, 30.18 mmol) and cesium carbonate (17.88 g, 54.88 mmol) were added successively. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, the resulting filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-29b (7.00 g, 24.53 mmol, yield 89.4%). MS-ESI [M+H] calculated: 286.1; found: 286.2. +

[0729] Second step

[0730] Intermediate B-2-29b (5.00 g, 17.52 mmol) was dissolved in tetrahydrofuran (50 mL). After being protected by nitrogen and cooled to -78 °C, 1N isopropyl magnesium bromide tetrahydrofuran solution (35.00 mL, 35.00 mmol) was slowly added dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed successively with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give two components.

[0731] The component 1 with shorter retention time was intermediate B-2-29c (1.20 g, 3.64 mmol, yield 20.8%). MS-ESI [M+H] calculated: 330.2; found: 330.2. +

[0732] The component 2 with longer retention time was intermediate B-2-30c (1.40 g, 4.25 mmol, yield 24.3%). MS-ESI [M+H] calculated: 330.2; found: 330.2. +

[0733] Third step - component 1

[0734] ​​​Intermediate B-2-29c (1.20 g, 3.64 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride in dioxane (10 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give crude intermediate B-2-29d which was used in the next step without purification.

[0735] Fourth step - Component 1

[0736] Crude intermediate B-2-29d was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (0.79 g, 3.91 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.02 g, 5.32 mmol), 1-hydroxybenzotriazole (0.72 g, 5.32 mmol) and ethyldiisopropylamine (1.38 g, 10.65 mmol) were added successively at -10 °C. After the addition, the reaction was continued for 2 h at -10 °C, water (10 mL) was added, and extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-29e (0.98 g, 2.40 mmol, 65.9% overall yield for two steps). MS-ESI Calcd for [M-Boc+2H] + : 309.2; Found: 309.1.

[0737] Fifth step - Component 1

[0738] Intermediate B-2-29e (0.98 g, 2.40 mmol) was dissolved in dichloromethane (5 mL), 4M hydrogen chloride in dioxane (10 mL) was added under ice-bath and the reaction was continued for 2 h, it was concentrated to give crude intermediate B-2-29f which was used in the next step without purification.

[0739] Sixth step - Component 1

[0740] Crude intermediate B-2-29f was dissolved in dichloromethane (10 mL), and acetyl chloride (0.21 g, 2.64 mmol) and ethyldiisopropylamine (0.93 g, 7.20 mmol) were added at -10 °C. After the reaction was continued for 1 h at -10 °C, water (10 mL) was added, and extracted with dichloromethane (10 mL x 3). The organic phases were combined, washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give product B-2-29 (0.45 g, 1.28 mmol, 53.5% overall yield for two steps). MS-ESI Calcd for [M+H] + : 351.2; Found: 351.3. 1H NMR (400 MHz, Chloroform-d) δ 8.57 (d, J = 8.7 Hz, 1H), 7.60 - 7.52 (m, 4H), 7.46 - 7.30 (m, 5H), 4.88 (dd, J = 9.5, 6.4 Hz, 1H), 4.77 (dd, J = 8.8, 6.9 Hz, 1H), 4.19 - 3.95 (m, 2H), 2.75 (ddt, J = 12.0, 9.0, 6.8 Hz, 1H), 2.43 - 2.28 (m, 1H), 2.10 - 2.00 (m, 1H), 1.97 (s, 3H), 0.90 (dd, J = 23.2, 6.7 Hz, 6H).

[0741] Third Step - Component 2

[0742] Intermediate B-2-30c (1.40 g, 4.25 mmol) was dissolved in dichloromethane (10 mL), and 4M hydrogen chloride in dioxane (10 mL) was added under ice bath. After the reaction was continued for 1 h under ice bath, it was concentrated to give crude intermediate B-2-30d which was used directly in the next step without purification.

[0743] Fourth Step - Component 2

[0744] Crude B-2-30d was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (0.88 g, 4.39 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.15 g, 5.99 mmol), 1-hydroxybenzotriazole (0.81 g, 5.99 mmol) and ethyldiisopropylamine (1.55 g, 11.97 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, and water (10 mL) was added, followed by extraction with dichloromethane (10 mL x 3). The organic phase was combined, washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-30e (0.80 g, 1.96 mmol, 46.1% overall yield for two steps). MS-ESI [M-Boc+2H] Calc’d: 309.2; Found: 308.9. +

[0745] Fifth Step - Component 2

[0746] Intermediate B-2-30e (0.80 g, 1.96 mmol) was dissolved in dichloromethane (3 mL), and 4M hydrogen chloride in dioxane (6 mL) was added under ice bath and the reaction was continued for 2 h. It was concentrated to give crude intermediate B-2-30f which was used directly in the next step without purification.

[0747] Sixth Step - Component 2​

[0748] The crude intermediate B-2-30f was dissolved in dichloromethane (5 mL), cooled to -10 °C, and acetyl chloride (0.17 g, 2.15 mmol) and ethyldiisopropylamine (0.76 g, 5.85 mmol) were added. After 1 h of reaction at -10 °C, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the product B-2-30 (0.49 g, 1.40 mmol, 71.3% total yield over two steps). MS-ESI Calcd for [M+H] + : 351.2; Found: 351.3. 1 H NMR (400 MHz, Chloroform-d) δ 8.63 (d, J = 9.0 Hz, 1H), 7.63 - 7.48 (m, 4H), 7.43 (dd, J = 8.4, 6.9 Hz, 2H), 7.35 - 7.30 (m, 1H), 7.28 (s, 1H), 7.26 (s, 1H), 4.90 (ddd, J = 61.6, 9.3, 6.6 Hz, 2H), 4.10 - 3.92 (m, 2H), 2.68 (ddt, J = 12.0, 9.4, 6.7 Hz, 1H), 2.39 (dtd, J = 12.0, 9.3, 5.5 Hz, 1H), 2.08 - 2.00 (m, 1H), 1.94 (s, 3H), 0.94 (dd, J = 30.7, 6.7 Hz, 6H).

[0749] Preparation of compounds B-2-31 and B-2-32

[0750]

[0751] First step

[0752] B-2-31a (4.00 g, 24.51 mmol) was dissolved in dichloromethane (50 mL), and (R)-tert-butylsulfinamide (2.97 g, 24.51 mmol) and cesium carbonate (15.97 g, 49.02 mmol) were added successively. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, it was filtered, the resulting filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-31b (6.53 g, 24.5 mmol, yield 100%). MS-ESI Calcd for [M+H] + : 267.1; Found: 267.2.

[0753] Second step

[0754] Intermediate B-2-31b (5.00 g, 18.77 mmol) was dissolved in tetrahydrofuran (50 mL). After nitrogen protection and cooling to -78 °C, 1 N isopropyl magnesium bromide tetrahydrofuran solution (37.5 mL, 37.5 mmol) was added dropwise slowly. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was completed. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed with saturated brine (100 mL x 2) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give two components.

[0755] The component 1 with shorter retention time was intermediate B-2-31c (1.12 g, 3.62 mmol, 19.2% yield). MS-ESI [M+H] calculated for C19H21NO4: 311.1; found: 311.2. +

[0756] The component 2 with longer retention time was intermediate B-2-32c (0.45 g, 1.45 mmol, 7.7% yield). MS-ESI [M+H] calculated for C19H21NO4: 311.1; found: 311.2. +

[0757] Third step - component 1

[0758] Intermediate B-2-31c (1.12 g, 3.62 mmol) was dissolved in dichloromethane (10 mL), and 4 M hydrogen chloride dioxane solution (10 mL) was added under ice bath. After the reaction was continued at ice bath for 1 h, it was concentrated to give intermediate B-2-31d crude product, which was used directly in the next step reaction without purification.

[0759] Fourth step - component 1

[0760] The intermediate B-2-31d crude product was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (0.75 g, 3.73 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.97 g, 5.08 mmol), 1-hydroxybenzotriazole (0.69 g, 5.08 mmol) and ethyldiisopropylamine (1.31 g, 10.17 mmol) were added successively at -10 °C. After addition, the reaction was continued at -10 °C for 2 h, then water (10 mL) was added, and extracted with dichloromethane (10 mL x 3). The organic phase was combined and washed with saturated brine (10 mL x 3) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-31e (0.90 g, 2.31 mmol, total yield of two steps 63.8%).

[0761] Fifth step - component 1 ​​

[0762] Intermediate B-2-31e (0.90 g, 2.31 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride solution in dioxane (10 mL) was added under ice-bath and the reaction was continued for 2 h, concentrated to give crude intermediate B-2-31f which was used in the next step without purification.

[0763] Step 6 - Component 1

[0764] The crude intermediate B-2-31f was dissolved in dichloromethane (10 mL), cooled to -10 °C, acetyl chloride (0.20 g, 2.55 mmol) and ethyl diisopropylamine (0.90 g, 6.96 mmol) were added. After 1 h reaction at -10 °C, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phase was combined and washed with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated, purified by silica gel column chromatography to give product B-2-31 (0.40 g, 1.21 mmol, 52.3% total yield over two steps). MS-ESI [M+H] calc’d for C19H21F2N4O2: 359.2; found: 359.2. + 1 HNMR (400 MHz, Chloroform-d) δ 8.73 (d, J = 8.4 Hz, 1H), 8.07 - 7.74 (m, 2H), 7.53 - 7.29 (m, 2H), 5.28 (dd, J = 8.4, 5.3 Hz, 1H), 4.99 (dd, J = 9.4, 6.4 Hz, 1H), 4.21 - 3.95 (m, 2H), 2.87 - 2.75 (m, 1H), 2.60 - 2.34 (m, 2H), 1.97 (s, 3H), 0.99 (dd, J = 22.7, 6.8 Hz, 6H).

[0765] Step 3 - Component 2

[0766] Intermediate B-2-32c (0.45 g, 1.45 mmol) was dissolved in dichloromethane (5 mL), 4M hydrogen chloride solution in dioxane (5 mL) was added under ice-bath. After 1 h reaction under ice-bath, concentrated to give crude intermediate B-2-32d which was used in the next step without purification.

[0767] Step 4 - Component 2

[0768] ​The crude B-2-32d was dissolved in dichloromethane (5 mL) and intermediate A-2-7b (0.32 g, 1.59 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.42 g, 2.17 mmol), 1-hydroxybenzotriazole (0.29 g, 2.17 mmol) and ethyldiisopropylamine (0.56 g, 4.35 mmol) were added successively at -10 °C. After the addition, the reaction was continued at -10 °C for 2 h, then water (5 mL) was added and extracted with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product was purified by column chromatography on silica gel to give intermediate B-2-32e (0.38 g, 0.98 mmol, 67.6% overall yield for two steps).

[0769] Fifth step - Component 2

[0770] Intermediate B-2-32e (0.38 g, 0.98 mmol) was dissolved in dichloromethane (5 mL) and 4 M hydrogen chloride in dioxane (5 mL) was added under ice bath and the reaction was continued for 2 h. The mixture was concentrated to give the crude intermediate B-2-32f which was used directly in the next step without purification.

[0771] Sixth step - Component 2

[0772] The crude intermediate B-2-32f was dissolved in dichloromethane (5 mL) and cooled to -10 °C. Acetyl chloride (80.0 mg, 1.02 mmol) and ethyldiisopropylamine (0.36 g, 2.79 mmol) were added. After the reaction was continued at -10 °C for 1 h, water (5 mL) was added and extracted with dichloromethane (5 mL x 3). The organic phases were combined and washed successively with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The product B-2-32 (71.4 mg, 0.22 mmol, 22.0% overall yield for two steps) was purified by column chromatography on silica gel. MS - ESI [M+H] calc’d for C19H22F2N4O4: 332.1; found: 332.2. + 1 ​HNMR (400 MHz, Chloroform-d) δ 8.71 (d, J = 8.7 Hz, 1H), 7.92 (ddt, J = 60.4, 8.0, 0.9 Hz, 2H), 7.46 (ddd, J = 8.3, 7.2, 1.3 Hz, 1H), 7.36 (ddd, J = 8.3, 7.2, 1.2 Hz, 1H), 5.33 (dd, J = 8.7, 5.2 Hz, 1H), 5.00 (dd, J = 9.6, 6.3 Hz, 1H), 4.14 - 4.06 (m, 2H), 2.78 - 2.59 (m, 1H), 2.62 - 2.34 (m, 2H), 2.01 (s, 3H), 1.03 (dd, J = 25.1, 6.8 Hz, 6H).

[0773] Preparation of compound B-2-33

[0774]

[0775] First step

[0776] B-2-33a (4.50 g, 31.00 mmol) was dissolved in dichloromethane (50 mL), (R)-tert-butylsulfinamide (4.13 g, 34.10 mmol) and cesium carbonate (20.20 g, 62.00 mmol) were added successively. After addition, the reaction was carried overnight, TLC showed that the reaction was complete, filtered, the obtained filtrate was washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-33b (7.50 g, 30.2 mmol, yield 97.4%). MS-ESI [M+H] calculated: 249.1; found: 249.1. +

[0777] Second step

[0778] Intermediate B-2-33b (5.00 g, 20.13 mmol) was dissolved in tetrahydrofuran (50 mL). After protection with nitrogen and cooling to -78 °C, 1 N isopropyl magnesium bromide tetrahydrofuran solution (40.26 mL, 40.26 mmol) was added slowly dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 3). The organic phase was combined and washed successively with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give a mixture of two components, recorded as intermediate B-2-33c (3.70 g, 12.7 mmol, yield 62.8%).

[0779] Third step ​

[0780] Intermediate B-2-33c (1.40 g, 4.79 mmol) was dissolved in dichloromethane (10 mL), and 4 M hydrogen chloride in dioxane (10 mL) was added under ice-bath. After the reaction was continued for 1 h under ice-bath, it was concentrated to give crude intermediate B-2-33d which was used directly in the next step without purification.

[0781] Fourth step

[0782] Crude intermediate B-2-33d was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (1.06 g, 5.26 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.37 g, 7.17 mmol), 1-hydroxybenzotriazole (0.97 g, 7.17 mmol) and ethyldiisopropylamine (1.85 g, 14.37 mmol) were added successively under -10 °C. After the addition, the reaction was continued for 2 h under -10 °C, and water (10 mL) was added, followed by extraction with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave intermediate B-2-33e (0.93 g, 2.50 mmol, 52.3% overall yield for two steps). MS-ESI Calcd for [M+H] + : 372.2; found: 372.3.

[0783] Fifth step

[0784] Intermediate B-2-33e (0.93 g, 2.50 mmol) was dissolved in dichloromethane (10 mL), and 4 M hydrogen chloride in dioxane (20 mL) was added under ice-bath and the reaction was continued for 2 h. It was concentrated to give crude intermediate B-2-33f which was used directly in the next step without purification.

[0785] Sixth step

[0786] Crude intermediate B-2-33f was dissolved in dichloromethane (10 mL), and acetyl chloride (0.22 g, 2.76 mmol) and ethyldiisopropylamine (0.97 g, 7.53 mmol) were added under -10 °C. After the reaction was continued for 1 h under -10 °C, water (10 mL) was added, followed by extraction with dichloromethane (10 mL x 3). The organic phases were combined and washed successively with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography gave product B-2-33 (272.5 mg, 0.87 mmol, 34.8% overall yield for two steps). MS-ESI Calcd for [M+H] + : 314.2; found: 314.3. 1H NMR (400 MHz, Chloroform-d) δ 8.91 (d, J = 21.2 Hz, 1H), 8.33 (dd, J = 53.4, 8.5 Hz, 1H), 7.55 (dt, J = 7.8, 1.0 Hz, 1H), 7.33 (ddq, J = 7.7, 5.8, 1.0 Hz, 1H), 7.19 - 6.99 (m, 2H), 6.34 (ddt, J = 14.5, 1.9, 0.8 Hz, 1H), 5.02 - 4.71 (m, 2H), 4.10 - 3.97 (m, 2H), 2.69 (dddd, J = 11.9, 9.0, 7.4, 6.3 Hz, 1H), 2.47 - 2.25 (m, 2H), 1.92 (d, J = 14.7 Hz, 3H), 1.01 (ddd, J = 23.0, 14.1, 6.8 Hz, 6H).

[0787] Preparation of compounds B-2-34 and B-2-35

[0788]

[0789] First step

[0790] B-2-34a (8.00 g, 43.24 mmol) was dissolved in dichloromethane (100 mL), (R)-tert-butylsulfinamide (5.24 g, 43.24 mmol) and cesium carbonate (28.18 g, 86.48 mmol) were added successively. After addition, the reaction was allowed to proceed overnight, TLC showed that the reaction was complete, water (100 mL) was added, and extracted with dichloromethane (100 mL x 2). The organic phase was combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-34b (10.13 g, 35.15 mmol, yield 81.3%). 1 H NMR (400 MHz, Chloroform-d) δ 8.46 (s, 1H), 7.66 - 7.62 (m, 2H), 7.56 - 7.52 (m, 2H), 1.19 (s, 9H).

[0791] Second step

[0792] Intermediate B-2-34b (8.00 g, 27.76 mmol) was dissolved in tetrahydrofuran (100 mL). After nitrogen protection and cooling to -78 °C, 1 N isopropyl magnesium bromide in tetrahydrofuran (50.0 mL, 50.0 mmol) was added slowly dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was completed. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 2). The organic phase was combined and washed with saturated brine (200 mL) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain two components.

[0793] The shorter retention time component 1 was intermediate B-2-34c (1.40 g, 4.21 mmol, 15.18% yield). MS-ESI [M+H] Theoretical: 332.1; Found: 332.2. + 1 H NMR (400 MHz, Chloroform-d) δ 7.44 - 7.36 (m, 2H), 7.14 - 7.02 (m, 2H), 4.05 - 4.01 (m, 1H), 3.35 (d, J = 5.9 Hz, 1H), 1.17 (s, 9H), 0.85 (d, J = 6.7 Hz, 3H), 0.72 (d, J = 6.7 Hz, 3H).

[0794] The longer retention time component 2 was intermediate B-2-35c (1.44 g, 4.34 mmol, 15.6% yield). MS-ESI [M+H] Theoretical: 332.1; Found: 332.2. + 1 H NMR (400 MHz, Chloroform-d) δ 7.42 - 7.33 (m, 2H), 7.09 - 7.03 (m, 2H), 4.06 (dt, J = 6.7, 1.7 Hz, 1H), 3.41 (d, J = 1.9 Hz, 1H), 1.12 (s, 9H), 0.90 (d, J = 6.7 Hz, 3H), 0.72 (d, J = 6.8 Hz, 3H).

[0795] Third step - component 1

[0796] Intermediate B-2-34c (1.40 g, 4.21 mmol) was dissolved in methanol (10 mL), and 4 M hydrogen chloride in ethyl acetate (5 mL) was added under ice bath. After being transferred to room temperature and continuing to react for 3 h, it was concentrated to obtain crude intermediate B-2-34d, which was used directly in the next step without purification.

[0797] Fourth step - component 1

[0798] ​​Intermediate A-2-7b (1.09 g, 5.41 mmol) was dissolved in dichloromethane (20 mL) and N, N-diisopropylethylamine (1.04 g, 5.41 mmol), 1-hydroxybenzotriazole (730.7 mg, 5.41 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.61 g, 12.48 mmol) were added successively at -10 °C. After 30 min of continuous reaction, the crude product of intermediate B-2-34d was added and the reaction was continued for 2 h, TLC showed that the reaction was completed. Water (20 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed successively with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated and purified by silica gel column chromatography to give intermediate B-2-34e (0.77 g, 1.87 mmol, 44.4% overall yield for two steps). MS-ESI [M-Boc+2H] + : 311.1; found: 311.2.

[0799] Fifth step - Component 1

[0800] Intermediate B-2-34e (0.77 g, 1.87 mmol) was dissolved in methanol (10 mL) and cooled to -5 °C, 4M hydrogen chloride ethyl acetate solution (10 mL) was added. It was transferred to room temperature and the reaction was continued for 2 h, concentrated to give the crude product of intermediate B-2-34f which was used directly in the next step without purification. Free amine MS-ESI [M+H] + : 311.1; found: 311.2.

[0801] Sixth step - Component 1

[0802] The crude product of intermediate B-2-34f was dissolved in dichloromethane (20 mL) and cooled to -10 °C, acetyl chloride (0.17 g, 2.16 mmol) and N, N-diisopropylethylamine (0.56 g, 4.32 mmol) were added. After 2 h of reaction at -10 °C, TLC showed that the reaction was completed. Water (30 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give product B-2-34 (496 mg, 1.40 mmol, 74.9% overall yield for two steps). MS-ESI [M+H] + : 353.1; found: 353.2. 1H NMR (400 MHz, Chloroform-d) δ 8.54 (d, J = 8.8 Hz, 1H), 7.35 (d, J = 8.4 Hz, 2H), 7.00 (d, J = 8.5 Hz, 2H), 4.88 (dd, J = 9.5, 6.5 Hz, 1H), 4.63 (dd, J = 8.8, 6.9 Hz, 1H), 4.04 - 3.85 (m, 2H), 2.61 - 2.52 (m, 1H), 2.37 - 2.26 (m, 1H), 1.86 (s, 3H), 0.86 (d, J = 6.7 Hz, 3H), 0.77 (d, J = 6.7 Hz, 3H).

[0803] Third Step - Component 2

[0804] Intermediate B-2-35c (1.44 g, 4.33 mmol) was dissolved in methanol (10 mL), 4M hydrogen chloride in ethyl acetate (5 mL) was added under ice-bath. It was continued to react for 3h at room temperature, TLC showed the reaction was completed. Concentration gave crude intermediate B-2-35d, which was used in the next step without purification.

[0805] Fourth Step - Component 2

[0806] Intermediate A-2-7b (1.09 g, 5.41 mmol) was dissolved in dichloromethane (20 mL), and N,N-diisopropyl ethylamine (1.04 g, 5.41 mmol), 1-hydroxybenzotriazole (730.7 mg, 5.41 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.61 g, 12.48 mmol) were added in turn at -10 °C. After the reaction was continued for 30 min, crude intermediate B-2-35d was added, and the reaction was continued for 2h, TLC showed the reaction was completed. Concentration, water (20 mL) was added, extracted with ethyl acetate (20 mL x 3). The organic phase was combined, washed with saturated brine (50 mL) in turn, dried over anhydrous sodium sulfate, concentrated, purified by silica gel column chromatography to give intermediate B-2-35e (1.00 g, 2.43 mmol, total yield of two steps 56.4%). MS-ESI [M-Boc+2H] Calc: 311.1; Found: 311.2. +

[0807] Fifth Step - Component 2

[0808] ​Intermediate B-2-35e (1.00, 2.43 mmol) was dissolved in methanol (10 mL), 4M hydrogen chloride in ethyl acetate (10 mL) was added under ice-bath and the reaction was continued for 2h, concentrated to give crude intermediate B-2-35f which was used in the next step without purification. MS-ESI [M+H]+calcd: 311.1, found: 311.2.

[0809] Step 6 - Component 2

[0810] The crude intermediate B-2-35f was dissolved in dichloromethane (20 mL), cooled to -10 °C, acetyl chloride (0.25 g, 3.24 mmol) and N,N-diisopropylethylamine (0.84 g, 6.48 mmol) were added. After reaction at -10 °C for 2h, TLC showed the reaction was complete. Water (30 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated, purified by silica gel column chromatography to give product B-2-35 (0.74 g, 2.10 mmol, 86.4% total yield over two steps). MS-ESI [M+H]+calcd: 353.1; found: 353.2. + 1 H NMR (400 MHz, Chloroform-d) δ 8.48 (d, J = 8.5 Hz, 1H), 7.36 (d, J = 8.5 Hz, 2H), 7.07 (d, J = 8.4 Hz, 2H), 4.78 (dd, J = 9.5, 6.5 Hz, 1H), 4.58 (dd, J = 8.5, 6.9 Hz, 1H), 4.00 (t, J = 7.8 Hz, 2H), 2.67 - 2.58 (m, 1H), 2.34 - 2.24 (m, 1H), 1.88 (s, 3H), 0.81 (d, J = 6.8 Hz, 3H), 0.75 (d, J = 6.7 Hz, 3H).

[0811] Preparation of compound B-2-36

[0812]

[0813] Step 1

[0814] ​B-2-36a (5.00 g, 41.61 mmol) was dissolved in dichloromethane (100 mL), (R)-tert-butylsulfinamide (5.04 g, 41.61 mmol) and cesium carbonate (27.11 g, 83.22 mmol) were added successively. After addition, the reaction was carried out for 2 h, TLC showed that the reaction was complete, water (100 mL) was added, and extraction was carried out with dichloromethane (100 mL x 2). The organic phase was combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain intermediate B-2-36b (8.79 g, 39.36 mmol, yield 94.6%). 1 H NMR (400 MHz, Chloroform-d) δ 8.47 (s, 1H), 7.70-7.63 (m, 2H), 7.19 (d, J = 7.9 Hz, 2H), 2.33 (s, 3H), 1.18 (s, 9H).

[0815] Second step

[0816] Intermediate B-2-36b (8.00 g, 35.82 mmol) was dissolved in tetrahydrofuran (50 mL). After being protected by nitrogen and cooled to -78 °C, 1 N isopropyl magnesium bromide tetrahydrofuran solution (70.0 mL, 70.0 mmol) was slowly added dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extraction was carried out with ethyl acetate (100 mL x 2). The organic phase was combined and washed successively with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain two components.

[0817] The component with shorter retention time was intermediate B-2-36c (2.01 g, 7.52 mmol, yield 21.0%). The other component was discarded.

[0818] Third step

[0819] Intermediate B-2-36c (2.00 g, 7.48 mmol) was dissolved in methanol (10 mL), and 4 M hydrogen chloride ethyl acetate solution (10 mL) was added under ice bath. After being transferred to room temperature and continuing to react for 3 h, it was concentrated to obtain crude intermediate B-2-36d, which was directly used in the next step reaction without purification.

[0820] Fourth step

[0821] Intermediate A-2-7b (2.33 g, 11.56 mmol) was dissolved in dichloromethane (20 mL) and N,N-diisopropylethylamine (2.99 g, 23.13 mmol), 1-hydroxybenzotriazole (1.35 g, 10.02 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.92 g, 10.02 mmol) were added successively at -10 °C. After 30 min of continuous reaction, the crude product of intermediate B-2-36d was added and the reaction was continued for 2 h, TLC showed that the reaction was completed. Water (20 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed successively with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated and purified by silica gel column chromatography to give intermediate B-2-36e (2.11 g, 6.09 mmol, 81.6% overall yield for two steps). MS-ESI [M-Boc+2H] + : 247.2; found: 247.3.

[0822] Fifth step

[0823] Intermediate B-2-36e (2.11 g, 6.09 mmol) was dissolved in methanol (10 mL) and cooled to -5 °C, 4M hydrogen chloride in ethyl acetate (10 mL) was added. It was transferred to room temperature and the reaction was continued for 2 h, concentrated to give the crude product of intermediate B-2-36f which was used directly in the next step without purification. Free amine MS-ESI [M+H] + : 247.2; found: 247.3.

[0824] Sixth step

[0825] The crude product of intermediate B-2-36f was dissolved in dichloromethane (20 mL) and cooled to -10 °C, acetyl chloride (0.62 g, 7.95 mmol) and N,N-diisopropylethylamine (2.05 g, 15.90 mmol) were added. After 2 h of reaction at -10 °C, TLC showed that the reaction was completed. Water (30 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give product B-2-36 (1.43 g, 4.96 mmol, 81.4% overall yield for two steps). MS-ESI [M+H] + : 289.2; found: 289.2. 1H NMR (400 MHz, Chloroform-d) δ 8.40 (d, J = 8.8 Hz, 1H), 7.08 (d, J = 8.2 Hz, 2H), 7.04 (d, J = 8.0 Hz, 2H), 4.77 (dd, J = 9.5, 6.4 Hz, 1H), 4.61 (dd, J = 8.8, 7.1 Hz, 1H), 4.03 - 3.96 (m, 2H), 2.66 (ddt, J = 11.9, 9.1, 6.7 Hz, 1H), 2.33 - 2.26 (m, 1H), 2.24 (s, 3H), 1.88 (s, 3H), 0.82 (d, J = 6.8 Hz, 3H), 0.75 (d, J = 6.7 Hz, 3H).

[0826] Preparation of compound B-2-37

[0827]

[0828] First step

[0829] B-2-37a (5.00 g, 52.04 mmol) was dissolved in dichloromethane (100 mL), (R)-tert-butylsulfinamide (6.31 g, 52.04 mmol) and cesium carbonate (33.91 g, 104.08 mmol) were added successively. After addition, the reaction was carried out for 2 h, TLC showed that the reaction was complete, water (100 mL) was added, and dichloromethane (100 mL x 2) was used for extraction. The organic phase was combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain intermediate B-2-37b (10.02 g, 50.28 mmol, yield 96.6%). + : 200.1, found: 200.2.

[0830] Second step

[0831] Intermediate B-2-37b (10.00 g, 27.76 mmol) was dissolved in tetrahydrofuran (100 mL). After being protected by nitrogen and cooled to -78 °C, 1N isopropyl magnesium bromide tetrahydrofuran solution (50.0 mL, 50.0 mmol) was slowly added dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 2). The organic phase was combined and washed successively with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain two components.

[0832] The shorter retention time was for intermediate B-2-37c (1.37 g, 7.68 mmol, 15.3% yield). MS-ESI [M+H] expected: 244.1; found: 244.3. The other fraction was discarded. + : 244.1; found: 244.3. The other fraction was discarded.

[0833] Third Step

[0834] Intermediate B-2-37c (1.87 g, 7.68 mmol) was dissolved in methanol (10 mL), and 4M hydrogen chloride in ethyl acetate (5 mL) was added under ice bath. It was transferred to room temperature and the reaction was continued for 3 h, then concentrated to give crude intermediate B-2-37d, which was used in the next step without purification.

[0835] Fourth Step

[0836] Intermediate A-2-7b (1.00 g, 4.14 mmol) was dissolved in dichloromethane (20 mL), and N,N-diisopropylethylamine (1.61 g, 12.45 mmol), 1-hydroxybenzotriazole (0.67 g, 4.98 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.95 g, 4.98 mmol) were added successively at -10 °C. The reaction was continued for 30 min, then crude intermediate B-2-37d was added and the reaction was continued for 2 h, TLC showed the reaction was completed. Water (20 mL) was added and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed successively with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated. Purification by silica gel column chromatography gave intermediate B-2-37e (0.91 g, 2.82 mmol, 36.7% yield for two steps). MS-ESI [M-Boc+2H] expected: 223.1; found: 223.3. + : 244.1; found: 244.3. The other fraction was discarded.

[0837] Fifth Step

[0838] Intermediate B-2-37e (0.91 g, 2.82 mmol) was dissolved in methanol (10 mL) and cooled to -5 °C, then 4M hydrogen chloride in ethyl acetate (10 mL) was added. It was transferred to room temperature and the reaction was continued for 2 h, then concentrated to give crude intermediate B-2-37f, which was used in the next step without purification.

[0839] Sixth Step

[0840] The crude intermediate B-2-37f was dissolved in dichloromethane (20 mL), cooled to -10 °C, acetyl chloride (0.36 mmol, 4.59 mmol) and N,N-diisopropyl ethylamine (1.14 g, 9.18 mmol) were added. After 2 h of reaction at -10 °C, TLC showed that the reaction was complete. Water (30 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the product B-2-37 (0.57 g, 2.16 mmol, 76.5% total yield over two steps). MS-ESI [M+H] Calc. for C15H17NO3: 265.1; Found: 265.4. + 1 H NMR (400 MHz, Chloroform-d) δ 8.30 (d, J = 9.1 Hz, 1H), 7.33 (dd, J = 1.9, 0.9 Hz, 1H), 6.28 (dd, J = 3.2, 1.8 Hz, 1H), 6.18 (d, J = 3.2 Hz, 1H), 4.88 (td, J = 9.6, 6.4 Hz, 2H), 4.11 - 4.04 (m, 2H), 2.73 (ddt, J = 11.9, 8.9, 6.8 Hz, 1H), 2.45 - 2.33 (m, 1H), 1.92 (s, 3H), 0.91 - 0.85 (m, 6H).

[0841] Preparation of compound B-2-38

[0842]

[0843] First step

[0844] B-2-38a (5.00 g, 44.19 mmol) was dissolved in dichloromethane (100 mL), (R)-tert-butylsulfinamide (5.36 g, 44.19 mmol) and cesium carbonate (28.80 g, 88.38 mmol) were added successively. After addition, the reaction was carried out for 2 h, TLC showed that the reaction was complete, water (100 mL) was added and extracted with dichloromethane (100 mL x 2). The organic phase was combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give the intermediate B-2-38b (8.27 g, 38.23 mmol, yield 86.5%).

[0845] Second step

[0846] ​Intermediate B-2-38b (5.00 g, 23.11 mmol) was dissolved in tetrahydrofuran (100 mL). After nitrogen protection and cooling to -78 °C, 1 N isopropyl magnesium bromide in tetrahydrofuran (50.0 mL, 50.0 mmol) was added slowly dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was completed. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 2). The organic phase was combined and washed with saturated brine (200 mL) successively, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give two components.

[0847] The component with shorter retention time was intermediate B-2-38c (2.51 g, 9.63 mmol, yield 41.7%). MS-ESI [M+H] calculated for C12H15NO2 + : 261.1; found: 261.1. 1 H NMR (400 MHz, Chloroform-d) δ 7.66 (d, J = 3.3 Hz, 1H), 7.21 (d, J = 3.2 Hz, 1H), 4.45 - 4.38 (m, 2H), 2.21 - 2.12 (m, 1H), 1.21 (s, 9H), 0.87 (dd, J = 6.8, 3.8 Hz, 6H).

[0848] The other component was discarded.

[0849] Third step

[0850] Intermediate B-2-38c (2.51 g, 9.63 mmol) was dissolved in methanol (10 mL), 4 M hydrogen chloride in ethyl acetate solution (5 mL) was added under ice bath. After being transferred to room temperature and continuing to react for 3 h, it was concentrated to give crude intermediate B-2-38d, which was used directly in the next step without purification.

[0851] Fourth step

[0852] Intermediate A-2-7b (1.31 g, 6.53 mmol) was dissolved in dichloromethane (50 mL) and N,N-diisopropylethylamine (2.11 g, 16.32 mmol), 1-hydroxybenzotriazole (0.88 g, 6.53 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.25 g, 6.53 mmol) were added successively at -10 °C. After the reaction was continued for 30 min, the crude product of intermediate B-2-38d was added and the reaction was continued for 2 h, TLC showed that the reaction was completed. Water (20 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed successively with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated and purified by silica gel column chromatography to give intermediate B-2-38e (1.38 g, 4.07 mmol, total yield of two steps 42.3%).

[0853] Fifth step

[0854] Intermediate B-2-38e (1.38 g, 4.07 mmol) was dissolved in methanol (10 mL) and cooled to -5 °C, 4M hydrogen chloride ethyl acetate solution (10 mL) was added. It was transferred to room temperature and the reaction was continued for 2 h, concentrated to give the crude product of intermediate B-2-38f which was used directly in the next step without purification.

[0855] Sixth step

[0856] The crude product of intermediate B-2-38f was dissolved in dichloromethane (20 mL) and cooled to -10 °C, acetyl chloride (0.48 g, 6.07 mmol) and N,N-diisopropylethylamine (1.57 g, 4.32 mmol) were added. After the reaction was continued for 2 h at -10 °C, TLC showed that the reaction was completed. Water (30 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give product B-2-38 (496.0 mg, 1.40 mmol, total yield of two steps 34.4%). MS-ESI [M+H] calc’d for C26H28N4O4: 282.2; found: 282.4. + 1 ​H NMR (400 MHz, Chloroform-d) δ 8.66 (d, J = 8.5 Hz, 1H), 7.75 (d, J = 3.3 Hz, 1H), 7.23 (d, J = 3.3 Hz, 1H), 5.21 (dd, J = 8.6, 5.4 Hz, 1H), 4.95 (dd, J = 9.5, 6.4 Hz, 1H), 4.14 - 4.07 (m, 2H), 2.82 - 2.70 (m, 1H), 2.46 - 2.38 (m, 2H), 1.96 (s, 3H), 0.96 (d, J = 6.8 Hz, 3H), 0.91 (d, J = 6.9 Hz, 3H).

[0857] Preparation of compounds B-2-39 and B-2-40

[0858]

[0859] First step

[0860] B-2-40a (5.00 g, 44.58 mmol) was dissolved in dichloromethane (100 mL), (R)-tert-butylsulfinamide (5.40 g, 44.58 mmol) and cesium carbonate (29.05 g, 89.16 mmol) were added successively. After addition, the reaction was carried out for 2 h, TLC showed that the reaction was complete, water (100 mL) was added, and dichloromethane (100 mL x 2) was used for extraction. The organic phase was combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain intermediate B-2-40b (7.87 g, 36.55 mmol, yield 92.0%). + : 216.0, found: 216.2.

[0861] Second step

[0862] Intermediate B-2-40b (5.00 g, 23.23 mmol) was dissolved in tetrahydrofuran (100 mL). After being protected by nitrogen and cooled to -78 °C, 2N isopropyl magnesium bromide tetrahydrofuran solution (24.0 mL, 48.0 mmol) was slowly added dropwise. After addition, the reaction was continued at -78 °C for 2 h, TLC showed that the reaction was complete. The pH was adjusted to 6 with saturated ammonium chloride solution, and extracted with ethyl acetate (100 mL x 2). The organic phase was combined and washed successively with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain two components.

[0863] The component 1 with shorter retention time was intermediate B-2-40c (2.51 g, 9.68 mmol, yield 41.7%).

[0864] The component 2 with longer retention time was intermediate B-2-39c (0.76 g, 2.90 mmol, yield 12.6%).

[0865] Third step - Component 1

[0866] Intermediate B-2-40c (1.12 g, 4.30 mmol) was dissolved in methanol (10 mL), 4M hydrogen chloride ethyl acetate solution (10 mL) was added under ice bath. It was transferred to room temperature and the reaction was continued for 3 h, then concentrated to give crude intermediate B-2-40d which was used directly in the next step without purification.

[0867] Fourth step - Component 1

[0868] Intermediate A-2-7b (671.1 mg, 3.36 mmol) was dissolved in dichloromethane (20 mL), and N,N-diisopropyl ethylamine (1.09 g, 8.41 mmol), 1-hydroxybenzotriazole (454.0 mg, 3.36 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (644.1 mg, 3.36 mmol) were added successively at -10 °C. After the reaction was continued for 30 min, crude intermediate B-2-40d was added and the reaction was continued for 2 h, TLC showed that the reaction was completed. Water (20 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed successively with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated, then purified by silica gel column chromatography to give intermediate B-2-40e (742.0 mg, 2.19 mmol, total yield of two steps 51.0%). MS-ESI [M-Boc+2H] Theoretical value: 239.1; Found: 239.2. + : 239.1; Found: 239.2.

[0869] Fifth step - Component 1

[0870] Intermediate B-2-40e (742.0 mg, 2.19 mmol) was dissolved in methanol (10 mL) and cooled to -5 °C, 4M hydrogen chloride ethyl acetate solution (10 mL) was added. It was transferred to room temperature and the reaction was continued for 2 h, then concentrated to give crude intermediate B-2-40f which was used directly in the next step without purification.

[0871] Sixth step - Component 1

[0872] The crude intermediate B-2-40f was dissolved in dichloromethane (20 mL), cooled to -10 °C, acetyl chloride (0.26 g, 3.29 mmol) and N,N-diisopropyl ethylamine (0.85 g, 6.57 mmol) were added. After 2 h of reaction at -10 °C, TLC showed the reaction was complete. Water (30 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give the product B-2-40 (468.0 mg, 1.67 mmol, 76.2% total yield for two steps). MS-ESI calc. for [M+H] + : 281.1; found: 281.2. 1 H NMR (400 MHz, Chloroform-d) δ 8.42 (d, J = 9.0 Hz, 1H), 7.16 (dd, J = 4.6, 1.6 Hz, 1H), 6.96 - 6.91 (m, 2H), 5.07 (dd, J = 8.9, 6.4 Hz, 1H), 4.87 (dd, J = 9.5, 6.4 Hz, 1H), 4.10 - 4.03 (m, 2H), 2.78 - 2.67 (m, 1H), 2.44 - 2.32 (m, 1H), 2.09 (h, J = 6.7 Hz, 1H), 1.94 (s, 3H), 0.93 (d, J = 2.0 Hz, 3H), 0.91 (d, J = 1.9 Hz, 3H).

[0873] Third Step - Component 2

[0874] The intermediate B-2-39c (0.76 g, 2.90 mmol) was dissolved in methanol (5 mL), 4M hydrogen chloride in ethyl acetate solution (5 mL) was added under ice bath. It was transferred to room temperature and continued to react for 3 h, then concentrated to give the crude intermediate B-2-39d, which was used directly in the next step without purification.

[0875] Fourth Step - Component 2

[0876] Intermediate A-2-7b (700 mg, 3.48 mmol) was dissolved in dichloromethane (50 mL) and N, N-diisopropylethylamine (1.12 g, 8.70 mmol), 1-hydroxybenzotriazole (0.47 g, 3.48 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (0.67 g, 3.48 mmol) were added successively at -10 °C. After 30 min of continuous reaction, the crude product of intermediate B-2-39d was added and the reaction was continued for 2 h, TLC showed the reaction was completed. Concentration, water (20 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (50 mL) successively, dried over anhydrous sodium sulfate, concentrated and purified by silica gel column chromatography to give intermediate B-2-39e (0.74 g, 2.19 mmol, 75.5% overall yield for two steps).

[0877] Fifth step - Component 2

[0878] Intermediate B-2-39e (0.74 g, 2.19 mmol) was dissolved in methanol (10 mL) and cooled to -5 °C, 4M hydrogen chloride in ethyl acetate solution (10 mL) was added. The temperature was transferred to room temperature and the reaction was continued for 2 h, concentrated to give the crude product of intermediate B-2-39f which was used directly in the next step without purification.

[0879] Sixth step - Component 2

[0880] The crude product of intermediate B-2-39f was dissolved in dichloromethane (20 mL) and cooled to -10 °C, acetyl chloride (0.26 g, 3.29 mmol) and N, N-diisopropylethylamine (0.85 g, 6.57 mmol) were added. After 2 h of reaction at -10 °C, TLC showed the reaction was completed. Water (30 mL) was added and extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give product B-2-39 (0.37 g, 1.32 mmol, 60.3% overall yield for two steps). MS-ESI Theoretical value [M+H]: 281.1; Found: 282.4. + 1 ​H NMR (400 MHz, Chloroform-d) δ 8.40 (d, J = 9.2 Hz, 1H), 7.09 (dd, J = 5.1, 1.2 Hz, 1H), 6.86 (dd, J = 5.1, 3.5 Hz, 1H), 6.80 (dt, J = 3.5, 1.1 Hz, 1H), 5.05 (dd, J = 8.9, 6.2 Hz, 1H), 4.87 (dd, J = 9.6, 6.4 Hz, 1H), 4.00 - 3.94 (m, 2H), 2.66 - 2.56 (m, 1H), 2.38 - 2.31 (m, 1H), 2.03 (h, J = 6.7 Hz, 1H), 1.87 (s, 3H), 0.88 (dd, J = 6.8, 5.5 Hz, 6H).

[0881] Preparation of compound B-2-41

[0882]

[0883] Intermediate B-2-3b (1.10 g, 4.93 mmol), N,N-diisopropylethylamine (2.55 g, 19.73 mmol) was dissolved in anhydrous dichloromethane (20 mL) and cooled to -10 °C. Acetyl chloride (0.77 g, 9.86 mmol) was added slowly dropwise. The reaction was stirred at 0 °C for 2 hours, TLC showed the reaction was complete. Water (50 mL) was added, extracted with ethyl acetate (100 mL x 3), the organic phase was combined, dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column chromatography to give the product B-2-41 (0.40 mg, 1.51 mmol, 30.6% yield). MS-ESI [M+H] calc’d for C17H22N2O4: 322.2; found: 322.2. + 1 H NMR (400 MHz, DMSO-d6) δ 8.76 (d, J = 8.6 Hz, 0.5H), 8.54 (d, J = 8.9 Hz, 0.5H), 8.10 - 8.03 (m, 1H), 7.18 (s, 1H), 4.94 - 4.67 (m, 2H), 4.03 (t, J = 7.7 Hz, 1H), 3.75 (dt, J = 9.0, 6.1 Hz, 1H), 2.49 - 2.32 (m, 1H), 2.25 - 1.95 (m, 2H), 1.77 (s, 1.5H), 1.57 (s, 1.5H), 0.89 (dd, J = 16.4, 6.7 Hz, 3H), 0.80 (dd, J = 8.6, 6.7 Hz, 3H).

[0884] Preparation of compound B-2-42

[0885] ​

[0886] First Step

[0887] Dissolve B-2-2a (10.00 g, 138.68 mmol) in dichloromethane (300 mL), add (S)-tert-butylsulfonamide (16.81 g, 138.68 mmol) and copper sulfate (5.53 g, 34.67 mmol). React for 16 h, TLC shows the reaction is complete. Filter, concentrate, dissolve in ethyl acetate (150 mL), slowly add petroleum ether (450 mL), stir for 12 h, filter, concentrate the filtrate to get the crude intermediate B-2-42a (15.00 g, 85.6 mmol, yield 61.7%).

[0888] Second Step

[0889] Under nitrogen protection, add oxazole (17.73 g, 256.71 mmol) to 1M borane tetrahydrofuran solution (171.14 mL, 171.14 mmol), and cool to -78°C. Slowly add 2.5M n-butyllithium tetrahydrofuran solution (171.14 mmol, 68.46 mmol), and continue to react for 30 min, then slowly add the crude intermediate B-2-42a (15.00 g, 85.6 mmol). Continue to react for 4 h, TLC shows the reaction is complete, pour the reaction into saturated aqueous ammonium chloride solution (200 mL), and extract with ethyl acetate (100 mL x 3). Combine the organic phases, wash with saturated brine (200 mL), dry over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography to get the intermediate B-2-42b (10.0 g, 40.9 mmol, yield 47.8%).

[0890] Third Step

[0891] Dissolve intermediate B-2-42b (10.0 g, 40.9 mmol) in methanol (200 mL), and add 4M hydrogen chloride dioxane solution (200 mL) at 0°C. Continue to react for 2 h, TLC shows the reaction is complete. Adjust the pH to 7 with ammonia water, concentrate. Dissolve the obtained residue in methanol (50 mL), add to stir for 1 h at 60°C, add acetonitrile (500 mL) and continue to stir for 1 h, slowly reduce to room temperature and precipitate the solid. Filter, dry to get the intermediate B-2-42c (5.00 g, 35.7 mmol, yield 87.2%). 1 H NMR (400 MHz, MeOD) δ 8.01 (s, 1H), 7.26 (s, 1H), 4.42 (d, J = 6.4 Hz, 1H), 2.45 - 2.22 (m, 1H), 1.09 (d, J = 6.8 Hz, 3H), 0.96 (d, J = 6.8 Hz, 3H).

[0892] Fourth Step

[0893] Intermediate A-2-6b (1.43 g, 7.11 mmol) was dissolved in dichloromethane (40 mL) and cooled to -10 °C. l-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.05 g, 10.70 mmol) and l-hydroxybenzotriazole (1.45 g, 10.70 mmol) were added. After 10 min of continued reaction, Intermediate B-2-42c (1.00 g, 7.14 mmol) and N,N-diisopropylethylamine (2.76 g, 21.39 mmol) were added. The reaction was continued for 3 h and TLC showed that the reaction was complete. Dichloromethane (50 mL) was added and washed with water (100 mL). The organic phase was collected and dried over anhydrous sodium sulfate, concentrated and purified by column chromatography on silica gel to give Intermediate B-2-42d (0.60 g, 1.86 mmol, 26.1% yield).

[0894] Fifth Step

[0895] Intermediate B-2-42d (0.60 g, 1.86 mmol) was dissolved in dichloromethane (25 mL) and 4 M hydrogen chloride in dioxane (5 mL) was added. After 1 h of reaction, TLC showed that the reaction was complete and it was concentrated to give crude Intermediate B-2-42e which was used directly in the next step without purification.

[0896] Sixth Step

[0897] Deuterated acetic acid (0.57 g, 8.96 mmol) was dissolved in N,N-dimethylformamide (20 mL) and 2-(7-azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.41 g, 8.96 mmol) was added. After 30 min of reaction, crude Intermediate B-2-42e and N,N-diisopropylethylamine (1.29 g, 10 mmol) were added sequentially. The reaction was continued for 16 h and TLC showed that the reaction was complete. Water (80 mL) was added and extracted with ethyl acetate (60 mL x 3). The organic phases were combined, washed with water (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography on silica gel to give product B-2-42 (45.1 mg, 0.17 mmol, 9.0% overall yield for two steps). MS-ESI Calcd for [M+H] + : 269.2; Found: 269.4. 1H NMR (400 MHz, CDC13) δ 8.37 (d, J = 8.2 Hz, 1H), 7.53 (s, 1H), 7.00 (s, 1H), 4.99 (dt, J = 19.6, 9.8 Hz, 1H), 4.87 (dd, J = 9.4, 6.6 Hz, 1H), 4.09 - 3.93 (m, 1H), 3.72 - 3.54 (m, 1H), 2.63 - 2.50 (m, 1H), 2.45 - 2.30 (m, 1H), 2.19 (td, J = 13.4, 6.8 Hz, 1H), 0.93 - 0.81 (m, 6H).

[0898] Preparation of compound B-2-43

[0899]

[0900] First step

[0901] Intermediate A-2-7b (1.43 g, 7.13 mmol) was dissolved in dichloromethane (40 mL) and cooled to -10 °C. l-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.05 g, 10.70 mmol) and l-hydroxybenzotriazole (1.45 g, 10.70 mmol) were added. After 10 min of reaction, intermediate B-2-42c (1.00 g, 7.14 mmol) and N,N diisopropylethylamine (2.76 g, 21.39 mmol) were added and the reaction was continued for 3 h. TLC showed that the reaction was complete. Dichloromethane (50 mL) was added and washed with water (100 mL). The organic phase was collected and dried over anhydrous sodium sulfate, concentrated and purified by column chromatography on silica gel to give intermediate B-2-43a (0.40 g, 1.24 mmol, 17.3% yield).

[0902] Second step

[0903] Intermediate B-2-43a (0.40 g, 1.24 mmol) was dissolved in dichloromethane (25 mL) and 4 M hydrogen chloride solution in dioxane (5 mL) was added. After 1 h of reaction, TLC showed that the reaction was complete and it was concentrated to give crude intermediate B-2-43b which was used directly in the next step without purification.

[0904] Third step

[0905] Dissolve deuterated acetic acid (0.20 g, 3.14 mmol) in N,N dimethylformamide (10 mL), add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.19 g, 3.14 mmol). After 30 min, add crude intermediate B-2-43b and N,N-diisopropylethylamine (0.61 g, 4.71 mmol) sequentially. Continue to react for 16 h, TLC shows the reaction is complete. Add water (80 mL), extract with ethyl acetate (60 mL x 3). Combine the organic phase, wash with water (100 mL), dry over anhydrous sodium sulfate, filter, concentrate, and purify by silica gel column chromatography to give product B-2-43 (78.2 mg, 0.29 mmol, 23.4% overall yield for two steps). MS-ESI calc. for [M+H] + : 269.2; found: 269.4. 1 H NMR (400 MHz, CDCl3) δ 8.50 (d, J = 7.7 Hz, 1H), 7.54 (d, J = 8.6 Hz, 1H), 7.01 (s, 1H), 4.98 (dd, J = 8.4, 5.9 Hz, 1H), 4.87 (dd, J = 9.3, 6.4 Hz, 1H), 4.10 - 3.93 (m, 2H), 2.79 - 2.58 (m, 1H), 2.43 - 2.29 (m, 1H), 2.18 (tt, J = 11.6, 5.8 Hz, 1H), 0.92 - 0.69 (m, 6H).

[0906] Preparation of compound B-2-44 of Preparation Example 71

[0907]

[0908] First step

[0909] A-2-38a (3.00 g, 13.81 mmol) was dissolved in methanol, 2-bromoethylamine hydrobromide (2.83 g, 13.81 mmol), 4-(4,6-dimethoxy-l,3,5-triazin-2-yl)-4-methyl morpholinium chloride (3.82 g, 13.81 mmol) and 4-methylmorpholine (1.40 g, 13.81 mmol) were added successively. After addition, the reaction was stirred at room temperature for 1 h, sodium hydroxide in methanol (2.71 g, 48.24 mmol, dissolved in 125 mL of methanol) was added, and the reaction was refluxed for 2.5 h. The reaction mixture was concentrated. Water (100 mL) was added to the residue, and the mixture was extracted with dichloromethane (100 mL x 3). The organic phases were combined and washed with saturated brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-44a (1.80 g, 7.43 mmol, 53.8% yield). MS-ESI [M+H] calc'd for C12H20N6O4: 243.2; found: 243.2. + : 243.2; found: 243.2.

[0910] Second Step

[0911] Intermediate B-2-44a (1.80 g, 7.43 mmol) was dissolved in dichloromethane (10 mL), and 4 M hydrogen chloride in dioxane (20 mL) was added under ice bath. After addition, the reaction was stirred for 2 h, and concentrated to give crude intermediate B-2-44b, which was used directly in the next step without purification. MS-ESI [M+H] calc'd for C7H10N4O2: 179.1; found: 179.1. + : 179.1; found: 179.1.

[0912] Third Step

[0913] Intermediate B-2-44b was dissolved in dichloromethane (10 mL), and intermediate A-2-7b (1.13 g, 5.60 mmol) was added. After cooling to -10 °C, N,N-diisopropylethylamine (2.17 g, 16.80 mmol), N-(3-dimethylaminopropyl)-N'- ethylcarbodiimide hydrochloride (1.40 g, 7.28 mmol) and 1-hydroxybenzotriazole (0.98 g, 7.28 mmol) were added, and the reaction was stirred for 3 h. The reaction mixture was concentrated, water (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give intermediate B-2-44c (1.80 g, 5.24 mmol, 70.5% yield for two steps). MS-ESI [M+H] calc'd for C21H26N8O4: 362.2; found: 362.4. + : 362.2; found: 362.4.

[0914] Fourth Step

[0915] Under ice-bath, intermediate B-2-44c (1.20 g, 3.32 mmol) was dissolved in dichloromethane (10 mL), 4M hydrogen chloride in dioxane (20 mL) was added, after 2 h, the reaction was concentrated to give crude intermediate B-2-44d, which was used in the next step without purification. MS-ESI [M+H] calculated: 262.1; found: 262.1. +

[0916] Fifth step

[0917] The crude intermediate B-2-44d was dissolved in dichloromethane (10 mL), cooled to -10 °C, N,N-diisopropylethylamine (1.48 g, 11.46 mmol) was added. Acetyl chloride (0.36 g, 4.58 mmol) was added slowly. The reaction was continued for 4 h, water (20 mL) was added, extracted with ethyl acetate (20 mL x 3). The organic phase was combined and washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give intermediate B-2-44e (0.80 g, 2.63 mmol, 79.3% total yield over two steps). MS-ESI [M+H] calculated: 304.1; found: 304.1. +

[0918] Sixth step

[0919] Intermediate B-2-44e (0.46 g, 1.5 mmol) was dissolved in methanol (10 mL), potassium hydroxide (0.35 g, 6.30 mmol) was added, heated to reflux for 2.5 h. After the reaction was completed, the solvent was rotary evaporated, water (10 mL) was added, extracted with ethyl acetate (10 mL x 3). The organic phase was combined and washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography to give intermediate B-2-44 (0.20 g, 0.75 mmol, 49.8% yield). MS-ESI [M+H] calculated: 268.2; found: 268.2. + 1 ​​​H NMR (400 MHz, Chloroform-d) δ 8.13 (dd, J = 47.5, 8.8 Hz, 1H), 4.84 (dt, J = 9.6, 6.6 Hz, 1H), 4.58 - 4.46 (m, 1H), 4.27 - 4.13 (m, 2H), 4.04 - 3.91 (m, 2H), 3.84 - 3.69 (m, 2H), 2.73 - 2.48 (m, 1H), 2.43 - 2.23 (m, 1H), 2.09 (qd, J = 6.9, 5.3 Hz, 1H), 1.86 (d, J = 4.0 Hz, 3H), 0.98 - 0.68 (m, 6H).

[0920] Comparative Example 1: Compound D4-pEL

[0921]

[0922] First Step

[0923] In a 100 mL three-necked flask, D4-pEL-1 (1.00 g, 4.36 mmol), L-leucine tert-butyl ester hydrochloride (0.98 g, 5.23 mmol), triethylamine (1.32 g, 13.08 mmol), N,N-dimethylformamide (20 mL), 1-hydroxybenzotriazole (1.18 g, 8.72 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.25 g, 6.54 mmol) were added successively. After the addition was completed, the reaction was allowed to proceed at room temperature for 5 hours. After the reaction was completed as indicated by LCMS and TLC monitoring, the reaction solution was diluted with saturated brine (100 mL), extracted with ethyl acetate (100 mL x 3), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by normal phase column chromatography. D4-pEL-3 (1.3 g, 3.26 mmol, 74.8% yield) was obtained. ESI-LCMS: m / z 243.2 [M+H]. t Bu-Boc + H + .

[0924] Second Step

[0925] In a 100 mL three-necked flask, D4-pEL-3 (1.3 g, 3.26 mmol) was added with a 1,4-dioxane solution of hydrochloric acid (4.0 M, 20 mL). The reaction solution was stirred at room temperature for 2 hours. After the reaction was completed as indicated by LCMS and TLC monitoring, the reaction solution was concentrated, diluted with dichloromethane (50 mL). Subsequently, the residual dioxane solution of hydrochloric acid was removed by concentration under reduced pressure. The residue was purified by multiple normal phase column chromatography. After lyophilization, D4-pEL (200 mg, 0.83 mmol, 25.5% yield) was obtained. ESI-LCMS: m / z 243.3 [M+H].+ .

[0926] 1 H NMR (400 MHz, DMSO-d6) δ 7.90 (s, 1H), 7.69 (d, J = 8.0 Hz, 1H), 4.08 - 3.91 (m, 2H), 2.29 - 1.97 (m, 3H), 1.94 - 1.83 (m, 1H), 1.69 - 1.55 (m, 1H), 1.55 - 1.46 (m, 1H), 1.46 - 1.35 (m, 1H), 0.84 (t, J = 6.4 Hz, 6H).

[0927] Comparative Example 2: Compound D5-PA

[0928]

[0929] D5-PA was purchased from Shanghai Bide Pharmaceutical Technology Co., Ltd., item number BD133842, batch number DSY151, purity 97%.

[0930] Experimental Example 1: Plasma stability determination

[0931] Mouse plasma (purchased from IPHASE, batch number 23B007) was thawed at 37°C water bath, and the supernatant was removed after centrifugation. The pH was recorded as 7-8. A certain volume of acetonitrile stock solution (concentration of 10 mM) of the test compound was diluted with acetonitrile to 1 mM as the test drug solution. A certain volume of the test drug solution was added to the corresponding volume of plasma and mixed uniformly, so that the concentration of the test compound in the incubation system was 5 μM, and the content of organic solvent acetonitrile was 0.5%. 50 μL of mixed plasma was taken in a 96-well plate (N = 2), and after incubation at 37°C water bath for a certain time, 300 μL of methanol solution containing internal standard was added to terminate the reaction, and the supernatant was vortexed and centrifuged for LC-MS / MS analysis. Propantheline and ZZL-7 were used as positive controls, and were treated and tested under the same conditions as the test compound. The percentage of compound remaining and incubation time were plotted to obtain k value, and the half-life of each compound was calculated. The formula is as follows:

[0932]

[0933] Table 1: Plasma stability data of some compounds

[0934]

[0935]

[0936]

[0937] Note: If the percentage of compound remaining is still greater than 75% after 120 minutes, T1 / 2 The retention time is > 289.1 min.

[0938] Table 1 shows the plasma stability data of some compounds, the results show that the compounds of the present application exhibit excellent plasma stability, which is significantly better than ZZL-7 and D5-PA.

[0939] Compound permeability detection of experimental example 2

[0940] MDR1-MDCK II cells (from the Netherlands Cancer Institute) were seeded in 96-well cell plates at a density of 3.33 x 10 5 Hank's balanced salt buffer (HBSS, pH 7.40 ± 0.05) containing 10.0 mM 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES) was used as the transport buffer. Compound DMSO stock solution was diluted to 2 μM (DMSO < 1.0%) with the transport buffer and added to the basal (B) and apical (A) ends (N = 2), respectively. The P-gp substrate digoxin was incubated at a concentration of 10 μM in both directions (A-B and B-A), and the low permeability control nado1ol and the high permeability control metoprolol were incubated at a concentration of 2 μM in one direction (A-B). After incubation in a cell incubator at 37 °C, 5% CO2 and saturated humidity for 2.5 hours, the final samples were collected from the receiving end and the dosing end, and the appropriate proportion of transport buffer and precipitant was added for LC-MS / MS analysis to calculate the apparent permeability coefficient, efflux ratio and solution recovery.

[0941] After the end of the transport experiment, the integrity of the MDR1-MDCK II cell layer was detected by using the fluorescent yellow detection experiment. The remaining solution in the apical and basal end wells was removed, 75 μL of transport buffer containing 100 μM fluorescent yellow was added to the apical well, and 250 μL of transport buffer was added to the basal end well. The cell plate was incubated in a cell incubator at 37 °C, 5% CO2 and saturated humidity for 30 minutes, then 20 μL of sample was taken from the apical end and mixed with 60 μL of transport buffer, and 80 μL of sample was taken from the basal end. The relative fluorescence intensity was detected by an enzyme-labeled instrument at 425 / 528 nm (excitation / emission) spectrum, and the fluorescent yellow penetration rate was calculated.

[0942] The apparent permeability coefficient (P app ), efflux ratio (Efflux Ratio, ER), and solution recovery (% Solution Recovery) were calculated using the following formulas, respectively:

[0943]

[0944] VR V is the volume of solution in the receiving end (0.075 and 0.25 mL for the apical and basal end sides, respectively), V D C is the volume of solution in the dosing end (0.075 and 0.25 mL for the apical and basal end sides, respectively), C R C is the peak area ratio of the compound in the receiving end, C D C is the peak area ratio of the compound in the dosing end, C0is the initial peak area ratio of the compound in the dosing end, Area is the surface area of the cell monolayer (0.143 cm 2 ), and Time is the incubation time 9000 s (150 min). P app (B-A) and P app (A-B) are the basal-to-apical apparent permeability coefficient and apical-to-basal apparent permeability coefficient, respectively.

[0945] Lucifer Yellow permeability (%Lucifer Yellow) was calculated using the following formula:

[0946]

[0947] RFU Apical and RFU Basolateral are the relative fluorescence intensity of Lucifer Yellow at the apical and basal end, respectively. V Apical and V Basolateral are the loading volume at the apical and basal end (0.075 and 0.25 mL, respectively).

[0948] Permeability data of some compounds in Table 2

[0949]

[0950] The compounds of the present patent are moderately permeable, better than the controls D4-pEL and D5-PA.

[0951] Experimental Example 3 Mouse pharmacokinetic study

[0952] The study was performed in 6-8 week old CD1 mice (purchased from: Sibeifu (Beijing) Biotechnology Co., Ltd.). The test compound was dissolved in 10% hydroxypropyl-β-cyclodextrin aqueous solution containing 10% DMSO, and administered by intravenous injection (10 mg / kg) or gavage (10 mg / kg). The compound concentration in the plasma sample was analyzed by LC-MS / MS method. Pharmacokinetic calculations were performed using WinNonlin (Phoenix™, version 8.3) or other similar software. The following pharmacokinetic parameters were calculated based on the plasma concentration versus time data: oral administration: AUC last , bioavailability (F). The above parameter data were statistically calculated.

[0953] Table 3 Mouse oral pharmacokinetic data of some compounds

[0954]

[0955]

[0956] Note: ZZL-7 dose is 100 mg / kg, PO.

[0957] Experimental Example 4 Rat pharmacokinetic test

[0958] The test was performed in 6-8 week old male SD rats (purchased from: Sibeifu (Beijing) Biotechnology Co., Ltd.). The test compound was dissolved in 10% DMSO + 15% Solutol HS + 75% (10% hydroxypropyl-β-cyclodextrin aqueous solution), and administered by intravenous injection (10 mg / kg) or gavage (15 mg / kg). The compound concentration in the plasma sample was analyzed by LC-MS / MS method. WinNonlin (Phoenix™, version 8.3) was used for pharmacokinetic calculation. The following pharmacokinetic parameters were calculated according to the plasma concentration and time data: oral administration: AUC last , bioavailability (F). The above parameter data were statistically calculated.

[0959] Table 4 Rat oral pharmacokinetic data of some compounds

[0960] Compound AUC last (h*ng / mL) F(％) B-2-3 24315 65.97 D4-pEL 98.6 1.04 D5-PA 11.5 0.71

[0961] Tables 3 and 4 show the area under the curve (AUC) and bioavailability (F) data of some compounds after oral administration, and the results show that the compounds of the present application have good pharmacokinetic properties, which are significantly better than ZZL-7, D4-pEL and D5-PA.

[0962] Experimental Example 5 In vivo pharmacodynamic test (1)

[0963] A 28-day animal chronic unpredictable mild stress model (CUMS, Chronic Unpredictable Mild Stress) was established in 6-8 week old C57 / B6 mice (purchased from: Jinan Pengyue Experimental Animal Breeding Co., Ltd.), and the experimental procedure was described in the literature [1,2] . The animals were adapted for one week, and seven experiments of fasting, foot electrostimulation, wet bedding, cage tilting, behavior restraint, shaking cage and olfactory stimulation were randomly performed within one week to achieve unpredictability, and the continuous stimulation lasted for 4 weeks. The open field test (Open Field Test, OFT) [3]After the success of the evaluation model, the efficacy is evaluated. The experiment is divided into a blank control group (Control), a model group (Model), a solvent control group (Vehicle), a fluoxetine control group, a ZZL-7 experimental group, and a compound group. The test compound is administered orally at a dose of 25 mg / kg, and the tail suspension test (TST) is performed 2 hours later [4,5] The depression of the mice is evaluated, and the prolonged immobility time of the mice in the TST indicates that the mice are in despair. After the data is collected and statistically analyzed, the SPSS data statistical software is used for analysis, and the Graph Pad software is used to draw images according to the SPSS analysis results.

[0964] The results of the tail suspension test are shown in Figure 1 and 2 The test results show that compounds B-2-3 and B-2-41 can quickly reverse the prolonged immobility time of mice in the tail suspension test in the chronic unpredictable stress model, proving that the compounds of the embodiments have a rapid antidepressant effect.

[0965] Example 6: In vivo efficacy detection (2)

[0966] 6-8-week-old C57 / B6 mice (purchased from: Jinan Pengyue Experimental Animal Breeding Co., Ltd.) are selected to establish a 28-day chronic unpredictable mild stress model (CUMS, Chronic Unpredictable Mild Stress), and the experimental procedure is described in the literature [1,2] The animals are adapted for one week, and seven experiments of fasting, water deprivation, foot electrostimulation, wet bedding, cage tilting, behavior restraint, shaking basket, and olfactory stimulation are randomly performed within one week to achieve unpredictability, and the continuous stimulation is performed for 4 weeks. The open field test (OFT) [3] After the success of the evaluation model, the efficacy is evaluated. The experiment is divided into a blank control group (Control), a model group (Model), a solvent control group (Vehicle), a fluoxetine control group, and a compound group. The test compound is administered orally at a dose of 25 mg / kg, and the tail suspension test (TST) is performed 2 hours later [4,5] The depression of the mice is evaluated, and the prolonged immobility time of the mice in the TST and FST indicates that the mice are in despair. After the data is collected and statistically analyzed, the SPSS data statistical software is used for analysis, and the Graph Pad software is used to draw images according to the SPSS analysis results.

[0967] The results of the forced swimming test are shown in Figure 3 , and the results of the tail suspension test are shown inFigure 4 The test results show that the compounds A-1-7, A-1-1 and A-2-1 can quickly reverse the prolongation of the immobile time of mice in the forced swimming and tail suspension tests in the chronic unpredictable stress model, proving that the embodiments of the present application have a rapid antidepressant effect.

[0968] Reference:

[0969] [1] Willner P, Towell A, Sampson D, Sophokleous S, Muscat R. Reduction of sucrose preference by chronic unpredictable mild stress, and its restoration by a tricyclic antidepressant. Psychopharmacology (Berl) 1987; 93: 358-64.

[0970] [2] Chen Z, Gu J, Lin S, et al. Saffron essential oil ameliorates CUMS-induced depression-like behavior in mice via the MAPK-CREB1-BDNF signaling pathway [J]. Journal of Ethnopharmacology, 2023, 300: 115719.

[0971] [3] Pentkowski, Nathan S., et al. "Anxiety and Alzheimer's disease: Behavioral analysis and neural basis in rodent models of Alzheimer's-related neuropathology." Neuroscience & Biobehavioral Reviews 127 (2021): 647-658.

[0972] [4] Can, Adem, et al. "The mouse forced swim test." JoVE (Journal of Visualized Experiments) 59 (2012): e3638.

[0973] [5] Can, Adem, et al. "The tail suspension test." JoVE (Journal of Visualized Experiments) 59 (2012): e3769.

Claims

1. A compound represented by formula (Ia-1’) or a pharmaceutically acceptable salt thereof, wherein, R1is selected from R6-(CO)-; R2is selected from 5-membered heteroaryl or phenyl, wherein the 5-membered heteroaryl or phenyl is optionally substituted with 0-8 R6; wherein the 5-membered heteroaryl is selected from: R3, R4are each independently selected from hydrogen or C1-C 15 alkyl; R3and R4are not simultaneously hydrogen; In case R1is selected from R6-(CO)-, R6is selected from hydrogen, C1-C 15 alkyl or C3-C6cycloalkyl, wherein C1-C 15 alkyl or C3-C6cycloalkyl is optionally substituted with 0 to 8 R 10 substituents; When R2 is selected from a 5-membered heteroaryl or phenyl group, wherein the 5-membered heteroaryl or phenyl group is optionally substituted by 0 to 8 R6 groups, R6 is selected from hydrogen, halogen, C1-C. 15 Alkyl or C1-C 15 alkoxy groups, where C1-C 15 Alkyl, C1-C 15 Alkyl groups are optionally surrounded by 0 to 8 R groups. 10 replace; R 10 selected from hydrogen, halogen, cyano or nitro.

2. The compound represented by formula (Ia-1’) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound represented by formula (Ia-1’) is a compound represented by formula (Ia-2) wherein R1, R2are as defined in claim 1.

3. The compound represented by formula (Ia-1’) or a pharmaceutically acceptable salt thereof according to claim 2, wherein the compound represented by formula (Ia-1’) is a compound represented by formula (Ia-3), wherein R2is as defined in claim 2.

4. The compound represented by formula (Ia-1’) or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein R2is selected from:

5. A compound or a pharmaceutically acceptable salt thereof, selected from any one of the following compounds:

6. A compound or a pharmaceutically acceptable salt thereof, selected from any one of the following compounds:

7. A pharmaceutical composition comprising a therapeutically effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, and a pharmaceutically acceptable adjuvant.

8. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, or a pharmaceutical composition according to claim 7, in the manufacture of a medicament for the prevention and / or treatment of depression.

9. Use according to claim 8, characterized in that, The medicament is a fast-acting medicament.

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