CN115260158A

CN115260158A - Compound of targeted protein regulator and application thereof

Info

Publication number: CN115260158A
Application number: CN202210476547.3A
Authority: CN
Inventors: 张朝欣; 毛海斌; 夏广新; 邓伟; 马星; 付建民; 王雪松; 付玉红; 柯樱; 王倩
Original assignee: Shanghai Pharmaceuticals Holding Co Ltd
Current assignee: Shanghai Pharmaceuticals Holding Co Ltd
Priority date: 2021-04-30
Filing date: 2022-04-29
Publication date: 2022-11-01
Also published as: WO2022228576A1

Abstract

The invention discloses a targetCompounds of protein modulators and uses thereof. The compound is shown as formula Ia, ib or Ic. The compound has good inhibition effect and protein degradation effect on KRAS _ G12D mutants.

Description

Compound of targeted protein regulator and application thereof

Technical Field

The invention relates to a compound of a targeted protein regulator and application thereof.

Background

RAS represents a group of closely related monomeric globular proteins of 189 amino acids (21 kDa molecular weight) that are associated with the plasma membrane and bind GDP or GTP. RAS acts as a molecular switch. When the RAS contains bound GDP, it is in the resting or closed position and is "inactive". In response to exposure of cells to certain growth-promoting stimuli, RAS is induced to exchange its bound GDP for GTP. In the case of GTP binding, RAS is "turned on" and is able to interact with other proteins (their "downstream targets") and activate the protein. The RAS protein itself has a very low intrinsic ability to hydrolyze GTP back to GDP, thereby turning itself into an off state. Turning RAS off requires an exogenous protein called Gtpase Activating Protein (GAP) that interacts with RAS and greatly accelerates the conversion of GTP to GDP. Any mutation in RAS that affects its ability to interact with GAPs or convert GTP back to GDP will result in prolonged protein activation and thus prolonged signaling to the cell telling it to continue growth and division. As these signals lead to cell growth and division, overactive RAS signaling can ultimately lead to cancer.

Structurally, the RAS protein contains a G domain responsible for the enzymatic activity of RAS, guanine nucleotide binding and hydrolysis (gtpase reaction). It also contains a C-terminal extension called CAAX box, which can be post-translationally modified and is responsible for targeting the protein to the membrane. The G domain is approximately 21-25kDa in size and contains a phosphate binding loop (P-loop). The P-loop represents the capsular bag of bound nucleotides in the protein, and this is a rigid part of the domain with conserved amino acid residues that are essential for nucleotide binding and hydrolysis (glycine 12, threonine 26 and lysine 16). The G domain also contains the so-called switch I (residues 30-40) and switch II (residues 60-76), which are both dynamic parts of the protein, often denoted as "spring-loaded" mechanisms due to the ability of the dynamic part to switch between resting and loaded states. The primary interaction is a hydrogen bond formed by threonine-35 and glycine-60 with the gamma-phosphate of GTP, which maintains the switch 1 and switch 2 regions, respectively, in their active conformations. After hydrolysis of GTP and release of phosphate, both relax into the inactive GDP conformation.

The most notable members of the RAS subfamily are HRAS, KRAS and NRAS, which are primarily implicated in many types of cancer. However, there are many other members, including DIRAS1; DIRAS2; DIRAS3; ERAS; a GEM; MRAS; NKIRAS1; NKIRAS2; NRAS; RALA; RALB; RAP1A; RAP1B; RAP2A; RAP2B; RAP2C; RASD1; RASD2; RASL10A; RASL10B; RASL11A; RASL11B; RASL12; REM1; REM2; RERG; RERGL; RRAD; RRAS and RRAS2.

Mutations in any of the three major isoforms of the RAS gene (HRAS, NRAS or KRAS) are among the most common events in human neoplasia. About 30% of all human tumors were found to carry some mutations in the RAS gene. Notably, KRAS mutations were detected in 25% -30% of tumors. In contrast, the rate of oncogenic mutations occurring in NRAS and HRAS family members is much lower (8% and 3%, respectively). The most common KRAS mutations were found at residues G12 and G13 and at residue Q61 in the P-loop. Among tumor-associated KRAS G12 mutations, KRAS G12D mutations occur with the highest probability of about 40%.

Based on the importance of aberrant activation of KRAS in cancer progression and the prevalence of KRAS gene mutations in human cancers, KRAS has been a target of interest to drug developers. Despite the advances made in this area, there remains a need in the art for improved inhibitors of KRAS G12D muteins.

In recent years, people utilize the functional characteristic that the ubiquitin proteasome pathway has specificity to degrade protein substrates to construct a target ubiquitination protein degradation chimeric molecule (PROTAC), wherein the PROTAC is a compound formed by connecting two ligands with different functions through a linker: one ligand targets the protein of interest (POI), while the other ligand specifically recruits E3 ligase. The ternary complex is formed when PROTAC binds to E3 ligase and the protein of interest, and by hijacking the E3 ligase, PROTAC presents a favorable spatial position to the POI to promote ubiquitination thereof, thereby selectively reducing the level of the target protein. The advantage of this approach is that ProTAC can catalyze multiple rounds of target protein degradation, which is the biggest difference between the PROTAC molecules and small molecule inhibitors.

CN110684015A discloses an ALK-targeting PROTAC molecule, and the ALK-targeting PROTAC molecule is successfully prepared, can effectively target a target protein, reduces the ALK content in cells, has better in vivo and in vitro antitumor activity, has lower toxicity to normal cells, and accords with the characteristics of high efficiency and low toxicity. However, no PROTAC molecules targeting KRAS G12D have been reported in the art.

Disclosure of Invention

The invention aims to overcome the defect that few types of compounds capable of performing targeted degradation on mutated KRAS in the prior art, and provides a PROTAC small molecular compound and application thereof. The compound has good inhibition effect on KRAS _ G12D mutant.

The present invention solves the above technical problems by the following technical solutions.

The invention provides a compound shown as formula Ia, ib or Ic, pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, metabolite or isotope compound thereof:

wherein R is ^1a 、R ^1b And R ^1c Independently is C ₆ -C ₁₈ Aryl radical, by R ^1a-1 Substituted C ₆ -C ₁₈ Aryl (R) ^1a-1 Is 1 or more, such as 1, 2 or 3) or 5 to 10 heteroaryl, the heteroatoms in said 5 to 10 heteroaryl being independently one or more of N, O or S, the number of heteroatoms being 1 to 4 (such as 1, 2, 3 or 4);

R ^1a-1 Independently is hydroxy, halogen, C ₁ -C ₁₂ Alkyl or C ₂ -C ₆ An alkynyl group;

R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c and R ^5c Independently is H orHalogen;

R ^3a is a 7-12 membered bridged heterocycloalkyl, wherein the heteroatoms in said bridged heterocycloalkyl are independently one or more of N, O, or S, and the number of heteroatoms is from 1 to 4 (e.g., 1, 2, 3, or 4);

R ^4c is H, 3-6 membered heterocycloalkyl or is-N (R) ^4c-1 R ^4c-2 ) Substituted 3-6 membered heterocycloalkyl (substituted with-N (R) ^4c- ¹ R ^4c-2 ) Number of substitution 1 or more, e.g. 1, 2 or 3), heteroatoms of said 3-to 6-membered heterocycloalkyl group and a-N (R) ^4c-1 R ^4c-2 ) The heteroatoms in the substituted 3-6 membered heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1-3 (e.g., 1, 2 or 3);

R ^4c-1 and R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

A ^a and A ^b Independently is CR ^5a Or N; r ^5a Is H, C ₁ -C ₄ Alkyl or halogen;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl (M) ^1-1 1 or more, such as 1, 2 or 3; m ^1-1 3-9 membered heterocycloalkyl which is substituted only), -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) -or absent (i.e., X) ^a Alkylene group of (1) and Y ¹ Directly connected); said 3-9 membered heterocycloalkyl group and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' the heteroatoms are independently one or more of N, O or S, and the number of heteroatoms is from 1 to 3 (e.g., 1, 2 or 3);

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ Alkyl or halogen；

Y ¹ Is O, -NR ⁸ Or is absent (i.e. M) ¹ And L ^a Directly connected);

the ring D is 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number of the heteroatoms is 1 or 2;

n2 is 0, 1, 2 or 3;

Y ² is O, -NR ⁹ Or is absent (i.e. X) ^a Alkylene group of (1) and L ^a Directly connected);

R ⁸ and R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 0, 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

the ring E is 3-6-membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number of the heteroatom is 1 or 2;

L ^a 、L ^b and L ^c Is a linking group;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand.

In a preferred embodiment, a compound represented by formula Ia, ib, or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof, or an isotopic compound thereof:

wherein R is ^1a 、R ^1b And R ^1c Independently is C ₆ -C ₁₈ Aryl radical, quilt R ^1a-1 Substituted C ₆ -C ₁₈ Aryl (R) ^1a-1 Is 1 or more, e.g. 1, 2 or 3) or 5-10 heteroaryl, wherein the heteroatoms in the 5-10 heteroaryl are independently one or more of N, O or S, and the number of heteroatoms is 1-4 (e.g., 1, 2, 3 or 4);

R ^1a-1 Independently is hydroxy, halogen or C ₁ -C ₁₂ An alkyl group;

R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c and R ^5c Independently is H or halogen;

R ^3a is a 7-12 membered bridged heterocycloalkyl group, wherein the heteroatoms in said bridged heterocycloalkyl group are independently one or more of N, O, or S, and the number of heteroatoms is 1-4 (e.g., 1, 2, 3, or 4);

R ^4c is H, 3-6 membered heterocycloalkyl or is-N (R) ^4c-1 R ^4c-2 ) Substituted 3-6 membered heterocycloalkyl (substituted with-N (R) ^4c- ¹ R ^4c-2 ) The number of substitution being 1 or more, e.g. 1, 2 or 3), the heteroatom of said 3-to 6-membered heterocycloalkyl group and the substituted radical-N (R) ^4c-1 R ^4c-2 ) The heteroatoms in the substituted 3-6 membered heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1-3 (e.g., 1, 2 or 3);

R ^4c-1 and R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl (M) ^1-1 1 or more, for example 1, 2 or 3; m is a group of ^1-1 3-9 membered heterocycloalkyl which is substituted only), -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) -or absence: (I.e. X ^a Alkylene group of (1) and Y ¹ Directly connected); said 3-9 membered heterocycloalkyl and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 The heteroatoms in' are independently one or more of N, O or S, and the number of heteroatoms is from 1 to 3 (e.g., 1, 2 or 3);

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ An alkyl group;

Y ¹ is O, -NR ⁸ Or is absent (i.e. M) ¹ And L ^a Directly connected);

the ring D is 3-6-membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number of the heteroatom is 1 or 2;

n2 is 0, 1, 2 or 3;

R ⁸ and R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 0, 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

L ^a 、L ^b and L ^c Is a linking group;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand.

In a preferred embodiment, the present invention provides a compound represented by formula Ia, ib or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof, or an isotopic compound thereof:

wherein R is ^1a 、R ^1b And R ^1c Independently is C ₆ -C ₁₈ Aryl radical, quilt R ^1a-1 Substituted C ₆ -C ₁₈ Aryl (R) ^1a-1 Is 1 or more, such as 1, 2 or 3) or 5 to 10 heteroaryl, the heteroatoms in said 5 to 10 heteroaryl being independently one or more of N, O or S, the number of heteroatoms being 1 to 4 (such as 1, 2, 3 or 4);

R ^1a-1 Independently is hydroxy, halogen, C ₁ -C ₁₂ Alkyl or C ₂ -C ₆ Alkynyl;

R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c and R ^5c Independently H or halogen;

R ^4c-1 and R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

A ^a and A ^b Independently of one another is CR ^5a Or N; r ^5a Is H, C ₁ -C ₄ Alkyl or halogen;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl (M) ^1-1 Is 1 or more, such as 1, 2 or 3; m is a group of ^1-1 3-9 membered heterocycloalkyl which is substituted only), -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) -or is absent (i.e. X) ^a Alkylene group of (1) and Y ¹ Directly connected); said 3-9 membered heterocycloalkyl group and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 The heteroatoms in' are independently one or more of N, O or S, and the number of heteroatoms is from 1 to 3 (e.g., 1, 2 or 3);

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ An alkyl group;

Y ¹ is O, -NR ⁸ Or is absent (i.e. M) ¹ And L ^a Directly connected);

n2 is 0, 1, 2 or 3;

R ⁸ and R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 0, 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

L ^a 、L ^b and L ^c Is a linking group;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand.

wherein R is ^1a 、R ^1b And R ^1c Independently is C ₆ -C ₁₈ Aryl radical, quilt R ^1a-1 Substituted C ₆ -C ₁₈ Aryl (R) ^1a-1 Is 1 or more, such as 1, 2 or 3) or 5 to 10 heteroaryl, the heteroatoms of said 5 to 10 heteroaryl being independently one or more of N, O or S, the number of heteroatoms being 1 to 4 (such as 1, 2, 3 or 4);

R ^1a-1 Independently is hydroxy, halogen or C ₁ -C ₁₂ An alkyl group;

R ^4c-1 and R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl (M) ^1-1 1 or more, for example 1, 2 or 3; m ^1-1 3-9 membered heterocycloalkyl which is substituted only), -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) -or absent (i.e., X) ^a Alkylene group of (1) and Y ¹ Directly connected); said 3-9 membered heterocycloalkyl group and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 The heteroatoms in' are independently one or more of N, O or S, and the number of heteroatoms is from 1 to 3 (e.g., 1, 2 or 3);

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ An alkyl group;

Y ¹ is O, -NR ⁸ Or is absent (i.e. M) ¹ And L ^a Directly connected);

n2 is 0, 1, 2 or 3;

R ⁸ and R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 0, 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

L ^a 、L ^b and L ^c Is a linking group;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand.

R ^1a-1 independently of one another is hydroxy, halogen or C ₁ -C ₁₂ An alkyl group;

R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c and R ^5c Independently H or halogen;

R ^4c is H, 3-6 membered heterocycloalkyl or is-N (R) ^4c-1 R ^4c-2 ) Substituted 3-6 membered heterocycloalkyl (substituted with-N (R) ^4c- ¹ R ^4c-2 ) Number of substitution is 1 or more, e.g. 1, 2 or 3), the heteroatom in said 3-6 membered heterocycloalkyl group and the substituted or unsubstituted-N (R) ^4c-1 R ^4c-2 ) The heteroatoms in the substituted 3-6 membered heterocycloalkyl are independently one or more of N, O or S, and the number of heteroatoms is 1-3 (e.g., 1, 2 or 3);

R ^4c-1 and R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

A ^a and A ^b Independently of one another is CR ^5a Or N; r is ^5a Is H, C ₁ -C ₄ Alkyl or halogen;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl (M) ^1-1 1 or more, for example 1, 2 or 3; m ^1-1 3-9 membered heterocycloalkyl which is substituted only), -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) -or is absent (i.e. X) ^a Alkylene group of (1) and Y ¹ Directly connected); said 3-9 membered heterocycloalkyl and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 The heteroatoms in' are independently one or more of N, O or S, and the number of heteroatoms is from 1 to 3 (e.g., 1, 2 or 3);

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ An alkyl group;

Y ¹ is O, -NR ⁸ Or is absent (i.e. M) ¹ And L ^a Directly connected);

n2 is 0, 1, 2 or 3;

R ⁸ and R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

the ring E is 3-6 membered heterocycloalkyl, the heteroatom in the heterocycloalkyl is N, and the number of the heteroatoms is 1 or 2;

L ^a 、L ^b and L ^c Is a linking group;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand.

In a preferred embodiment, R ^1a 、R ^1b And R ^1c In (b), the C ₆ -C ₁₈ Aryl or said aryl is substituted by R ^1a-1 Substituted C ₆ -C ₁₈ C in aryl ₆ -C ₁₈ Aryl is phenyl, naphthyl, phenanthryl or anthracyl, preferably naphthyl.

In a preferred embodiment, when R ^1a 、R ^1b And R ^1c Independently is a quilt R ^1a-1 Substituted C ₆ -C ₁₈ Aryl, said is R ^1a-1 Substituted C ₆ -C ₁₈ Aryl is

Wherein R is ⁶ And R ⁷ Independently H, halogen (e.g. F, cl, br or I), C ₁ -C ₁₂ Alkyl (e.g. methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl) or C ₂ -C ₆ Alkynyl (e.g. C) ₂ -C ₄ Alkynyl, for example ethynyl).

In a preferred embodiment, when R ^1a 、R ^1b And R ^1c Independently is

While said

Is composed of

In a preferred embodiment, when R ^1a 、R ^1b And R ^1c Independently is

When it is used, the

Is composed of

In a preferred embodiment, when R ^1a 、R ^1b And R ^1c Independently is

When it is used, the

Is composed of

In a preferred embodiment, R ⁶ 、R ⁷ 、R ^1a 、R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c 、R ^5a And R ^5c Wherein said halogen is F, cl, br or I, such as F or Cl.

In a preferred embodiment, R ⁶ 、R ⁷ And R ^1a In (b), the C ₁ -C ₁₂ Alkyl is C ₁ -C ₆ The alkyl group may be a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group or a tert-butyl group.

In a preferred embodiment, R ⁶ 、R ⁷ And R ^1a In (b), the C ₂ -C ₆ Alkynyl is C ₂ -C ₄ The alkynyl group may be an ethynyl group.

In a preferred embodiment, R ^3a Wherein the 7-12 membered bridged heterocycloalkyl is 7-or 8-membered bridged heterocycloalkyl, the heteroatom is N, the number of the heteroatoms is 2, and the heterocycle may be diazabicyclo [ 2.2.1%]Heptylalkyl, diazabicyclo [3.2.1]Octyl or diazabicyclo [2.2.2 ]]An octyl group.

In a preferred embodiment, R ^3a Wherein said 7-12 membered bridged heterocycloalkyl is attached to the pyrimidine ring through a heteroatom.

In a preferred embodiment, R ^3a Wherein said 7-12 membered bridged heterocycloalkyl group is

(e.g. in

)、

(e.g. in the case of

) Or

In a preferred embodiment, R ^4c Wherein said 3-to 6-membered heterocycloalkyl group is substituted with-N (C)R ^4c-1 R ^4c-2 ) The 3-6 membered heterocycloalkyl group of the substituted 3-6 membered heterocycloalkyl group is azetidinyl, pyrrolidinyl or piperidinyl.

In a preferred embodiment, R ^4c Wherein said 3-to 6-membered heterocycloalkyl group is substituted with said-N (R) ^4c-1 R ^4c-2 ) The 3-6 membered heterocycloalkyl group of the substituted 3-6 membered heterocycloalkyl group is attached to the pyrimidine ring through a heteroatom.

In a preferred embodiment, R ^4c-1 And R ^4c-2 In (b), the C ₁ -C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

In a preferred embodiment, R ^4c In (c), the quilt is-N (R) ^4c-1 R ^4c-2 ) Substituted 3-6 membered heterocycloalkyl is substituted by 1-N (CH) ₃ ) ₂ ) Substituted azetidinyl radicals, e.g.

In a preferred embodiment, M ¹ Wherein said 3-9 membered heterocycloalkyl is 3-6 membered monocyclic heterocycloalkyl, the heteroatoms are independently N and/or O, the number of heteroatoms is 1 or 2, and further is pyrrolidinyl (e.g., pyrrolidinyl)

The a terminal being linked to the alkylene radical and the b terminal being linked to Y ¹ Connected).

In a preferred embodiment, M ¹ In (1), the quilt M ^1-1 The 3-9 membered heterocycloalkyl group in the substituted 3-9 membered heterocycloalkyl group is an 8-9 membered fused ring hybrid alkyl group or a 3-6 membered monocyclic heterocycloalkyl group, the heteroatoms are independently N and/or O, the number of heteroatoms is 1 or 2, and further is a pyrrolidinyl group or a hexahydro-1H-pyrrolizinyl group.

In a preferred embodiment, M ^1-1 In (b), the C ₁ -C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl.

A certain oneIn a preferred embodiment, M ^1-1 In (1), the halogen is F, cl, br or I, such as F.

In a preferred embodiment, M ¹ In (2), the quilt M ^1-1 Substituted 3-9 membered heterocycloalkyl is 3-6 membered monocyclic heterocycloalkyl substituted by methyl, the heteroatoms are independently N and/or O, the number of heteroatoms is 1 or 2, further methyl substituted pyrrolidinyl (e.g. methyl substituted pyrrolidinyl)

The a terminal being linked to the alkylene group and the b terminal being linked to Y ¹ Connected).

In a preferred embodiment, M ¹ In (1), the quilt M ^1-1 Substituted 3-9 membered heterocycloalkyl is hexahydro-1H-pyrrolizinyl substituted by F, further

The a terminal being linked to the alkylene group and the b terminal being linked to Y ¹ Are connected.

In a preferred embodiment, M ¹ In (b), the-3-to 9-membered heterocycloalkyl- (CH) ₂ ) _n1’ The 3-9 membered heterocycloalkyl in OC (= O) -is an 8-9 membered fused ring hybrid alkyl group, the heteroatoms are independently N and/or O, the number of heteroatoms is 1 or 2, and further is

(a terminal and

alkylene group of (i.e., - (CH)) ₂ ) _n1 -) connected and the a' end is connected with a-3-9 membered heterocycloalkyl- (CH) ₂ ) _n1‘ Alkylene in OC (= O) - (CH) ₂ ) _n1’ -) are connected.

In a preferred embodiment, the 3-6 membered heterocycloalkyl group in ring D is azetidinyl (e.g., azetidinyl)

) Pyrrolidinyl (e.g. phenyl)

) Or piperazine (e.g. of the formula

)。

In a preferred embodiment, R ^5a 、R ⁸ 、R ⁹ 、R ¹⁰ 、R ¹¹ And M ^1-1 In (b), the C ₁ -C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl or ethyl.

In a preferred embodiment, the 3-6 membered heterocycloalkyl in ring E is azetidinyl (e.g., azetidinyl)

)。

In a preferred embodiment, L ^a 、L ^b And L ^c Independently is

e terminal and X ^a 、X ^b Or X ^c Connected with terminal f and Q ^a 、Q ^b Or Q ^c Connecting;

n4, n6 and n7 are independently 0, 1 or 2;

n5 and n8 are independently any integer from 1 to 5 (e.g., 1, 2, 3, 4, or 5);

Y ³ is NH, CH ₂ Or is absent;

n9 is any integer from 0 to 13 (e.g., 0, 1, 2, or 12);

n10 and n13 are independently 0, 1 or 2;

n11 and n12 are independently any integer from 1 to 7 (e.g., 1, 2, 3, 4, or 5);

z is CH ₂ Or N;

Y ⁴ is-C (= O) or absent;

Y ⁵ is 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl or 5-6 membered heteroaryl, the heteroatoms in the 3-6 membered heterocycloalkyl are independently one or more of N, O or S, the number of heteroatoms is 1, 2 or 3, the heteroatoms in the 5-6 membered heterocycloalkyl are independently one or more of N, O or S, the number of heteroatoms is 1, 2 or 3;

n14 and n15 are independently 0, 1,2,3, 4 or 5;

n16 and n19 are 0, 1 or 2;

n17, n18 and n20 are independently 0, 1,2,3, 4 or 5;

Z ¹ is CH or N.

Y ⁵ The 3-to 6-membered cycloalkyl group is preferably a cyclopropyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group, for example, a cyclohexyl group.

Y ⁵ The 3-to 6-membered heterocycloalkyl group in (1) is preferably a 5-membered heterocycloalkyl group having O or S as a hetero atom, for example, furyl

Y ⁵ The 5-to 6-membered heteroaryl group is preferably a 5-membered heteroaryl group having N as a heteroatom and 3 in number, e.g. 1-H,1,2, 3-triazolyl, e.g.

In a preferred embodiment, L ^a 、L ^b And L ^c Independently is

n4, n6 and n7 are independently 0, 1 or 2;

n5 and n8 are independently any integer from 1 to 5 (e.g., 1,2,3, 4, or 5);

Y ³ is NH, CH ₂ Or is absent;

n9 is any integer from 0 to 13 (e.g., 0, 1,2, or 12);

n10 and n13 are independently 0, 1 or 2;

n11 and n12 are independently any integer from 1 to 7 (e.g., 2,3, 4, or 5);

z is CH ₂ Or N;

Y ⁴ is-C (= O) or absent.

In a preferred embodiment, L ^a 、L ^b And L ^c Independently is

Wherein the left end is connected with X ^a (X ^b Or N in piperazinyl) and the right end is connected to Q ^a (Q ^b Or Q ^c ) Are connected.

In a preferred embodiment, L ^a 、L ^b And L ^c Independently is

In a preferred embodiment, Q ^a 、Q ^b And Q ^c Independently is

In a preferred embodiment, R ⁷ Is H or halogen (e.g., F).

In a preferred embodiment, R ^2a 、R ^2b 、R ^2c And R ^5c Independently a halogen (e.g., F).

In a preferred embodiment, when A ^a And A ^b Independently is CR ^5a When R is ^5a Independently a halogen (e.g., cl).

In a preferred embodiment, X ^a Is composed of

In a preferred embodiment, X ^a Is composed of

(e.g. in the case of

)、

(e.g. in

)、

(e.g. in

) Or

(e.g. in

)。

In a preferred embodiment, n1 is 1 or 3.

In a certain preferred directionIn the table, X ^b Is composed of

In a preferred embodiment, n2 is 0 or 1.

In a preferred embodiment of the present invention,

is composed of

M ¹ Is absent or 3-9 membered heterocycloalkyl (e.g.

)，Y ¹ is-NR ⁸ 。

In a preferred embodiment of the present invention,

is composed of

M ¹ Is as a quilt M ^1-1 Substituted 3-9 membered heterocycloalkyl (e.g. phenyl)

)，Y ¹ Is O.

In a preferred embodiment of the method of the present invention,

is composed of

M ¹ Is 3-9 membered heterocycloalkyl (e.g.

)，Y ¹ Is absent.

In a preferred embodiment of the method of the present invention,

is composed of

M ¹ Is-3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 OC (= O) -or absent, Y ¹ is-NR ⁸ ；M ¹ Is 3-9 membered heterocycloalkyl, Y ¹ Is absent; or, M ¹ Is as a quilt M ^1-1 Substituted 3-9 membered heterocycloalkyl, Y ¹ Is O.

In a preferred embodiment of the present invention,

is composed of

In a preferred embodiment of the present invention,

is composed of

In a preferred embodiment of the method of the present invention,

is composed of

In a preferred embodiment, the compound of formula Ia has the formula Ia-1:

Wherein R is ^2a Is halogen;

R ^3a is 7-12 membered bridged heterocycloalkyl, the 7-12 membered bridged heterocycloalkyl is 7-or 8-membered bridged heterocycloalkyl, the heteroatom is N, the number is 2,

R ^5a independently is halogen;

X ^a is composed of

M ¹ Is-3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 OC (= O) -or absent, Y ¹ is-NR ⁸ (ii) a Or, M ¹ Is 3-9 membered heterocycloalkyl or by M ^1-1 Substituted 3-to 9-membered heterocycloalkyl, Y ¹ Is O or absent;

L ^a is composed of

In a preferred embodiment, the compound of formula Ia has the formula Ia-1:

wherein R is ^2a Is halogen;

R ^5a is halogen;

X ^a is composed of

L ^a Is composed of

In a preferred embodiment, the compound of formula Ia has the formula Ia-3:

wherein R is ^2a Is halogen;

R ^5a is halogen;

X ^a is composed of

M ¹ Is 3-9 membered heterocycloalkyl or by M ^1-1 Substituted 3-9 membered heterocycloalkyl;

Y ¹ is absent;

L ^a is composed of

In a preferred embodiment, the compound represented by formula Ia-1 is Ia-1-1 or Ia-1-2:

R ^5a independently C ₁ -C ₄ Alkyl or halogen.

In a preferred embodiment, the compound represented by formula Ib has a general formula of formula Ib-1:

wherein R is ^5a Is halogen;

R ^2b is halogen;

X ^b is composed of

n3' is 0;

L ^b is composed of

n7 is 0, n8 is 3, 4 or 5 ³ Is CH ₂ Or is absent;

n10 and n13 are 0, n11 and n12 are independently 3, 4 or 5 ⁴ Is absent;

wherein R is ^5a Is halogen;

R ^2b is halogen;

X ^b is composed of

In a preferable embodiment, the compound shown in formula Ib-1 has a general formula Ib-1-1 or Ib-1-2:

in a preferred embodiment, the compound represented by formula Ic has the formula Ic' or Ic ":

preferably, in said formula Ic' or Ic ":

R ^2a and R ^5c Independently is halogen;

L ^c is composed of

n4 and n6 are 2, n5 is 4;

R ^4c is represented by-N (R) ^4c-1 R ^4c-2 ) Substituted 3-6 membered heterocycloalkyl.

In a preferred embodiment, the compound of formula Ia has the formula Ia-2:

wherein R is ^2a Is halogen; r ^5a Is halogen.

In a preferred embodiment, the compound represented by formula Ia, ib or Ic is any one of the following compounds:

Or

Represents a chemical bond in an axial chiral stereoconfiguration, and the 'x' represents that a chiral carbon atom is in an S configuration or an R configuration.

By passing

After the reaction, a compound was obtained, the retention time of the compound 40-5a was 8.91 minutes under the following chiral conditions: chiral column CHIRALPAK IC,2 × 25 cm, 5 microns; mobile phase A: n-hexane (10 mmol/l ammonia methanol), mobile phase B: ethanol; flow rate: 20 ml/min; elution was performed with 10% phase B in 21 minutes, detector UV 230/210 nm;

by passing

The reaction yielded compound 46-2a with a retention time of 4.85 minutes under chiral conditions as follows: CHIRAL column CHIRAL ART Cellulose-SC,3 × 25 cm, 5 μm; mobile phase A: supercritical carbon dioxide, mobile phase B: ethanol (0.5%, 2 mol/l ammonia methanol); flow rate: 60 ml/min; column temperature: 35 ℃ is carried out; eluting with 35% mobile phase B; detector UV222 nm;

by passing

The reaction yielded a compound with a retention time of 2.00 minutes for compound 47-1a under the following chiral resolution conditions: chiral column CHIRALPAK AD-H,3 × 25 cm, 5 microns; mobile phase A: supercritical carbon dioxide, mobile phase B: isopropanol (0.5%, 2 mol/l ammonia methanol); flow rate: 50 ml/min; column temperature: 35 ℃ is carried out; eluting with 30% mobile phase B; detector UV222 nm.

In a certain scheme, the salt form of the pharmaceutically acceptable salt of the compound shown in formula Ia, ib or Ic is formate, trifluoroacetate or hydrochloride.

The number of the salts in the pharmaceutically acceptable salts of the compounds shown in the formulas Ia, ib or Ic is preferably 1, 2, 3, 4, 5 or 6.

In a certain embodiment, the pharmaceutically acceptable salt of the compound represented by formula Ia, ib or Ic is any one of the following compounds:

or

Represents a chemical bond with a steric configuration, "+" represents that the chiral carbon atom is in an S configuration or an R configuration.

In a certain embodiment, the compound represented by formula Ia, ib or Ic, or a pharmaceutically acceptable salt thereof is any one of the following compounds:

means a chemical bond representing an axial chiral stereoconfiguration

Or

"x" indicates that the chiral carbon atom is in the R configuration or S configuration.

By passing

After the reaction, the formate of the compound was obtained, and the retention time of the compound 40-5a under the following chiral conditions was 8.91 minutes: chiral column CHIRALPAK IC,2 × 25 cm, 5 microns; mobile phase A: n-hexane (10 mmol/l ammonia methanol), mobile phase B: ethanol; flow rate: 20 ml/min; elution was performed with 10% phase B in 21 minutes, detector UV 230/210 nm;

by passing

The hydrochloride salt of the compound obtained from the reaction, compound 46-2a, had a retention time of 4.85 minutes under chiral conditions as follows: CHIRAL column CHIRAL ART Cellulose-SC,3 × 25 cm, 5 μm; mobile phase A: supercritical carbon dioxide, mobile phase B: ethanol (0.5%, 2 mol/l ammonia methanol); flow rate: 60 ml/min; column temperature: 35 ℃ is carried out; eluting with 35% mobile phase B; detector UV222 nm;

by passing

The reaction yielded the formate salt of compound, compound 47-1a, with a retention time of 2.00 minutes under the following chiral resolution conditions: chiral column CHIRALPAK AD-H,3 × 25 cm, 5 microns; a mobile phase A: supercritical carbon dioxide, mobile phase B: isopropanol (0.5%, 2 mol/l ammonia methanol); flow rate: 50 ml/min; column temperature: 35 ℃; elution with 30% mobile phase B; detector UV222 nm.

The above-mentioned compounds represented by formula Ia, ib or Ic, pharmaceutically acceptable salts thereof, solvates thereof, stereoisomers thereof, tautomers thereof, prodrugs thereof, metabolites thereof or isotopic compounds thereof of the present invention can be synthesized by methods comprising methods similar to those well known in the chemical art, and the steps and conditions thereof can be synthesized by referring to the steps and conditions of analogous reactions in the art, particularly according to the description herein. The starting materials are generally from commercial sources, such as Aldrich or can be readily prepared using methods well known to those skilled in the art (obtained via SciFinder, reaxys online databases).

The invention provides a compound shown in formula IIa or IIb:

wherein R is ^12a And R ^12b Independently a hydroxy protecting group (e.g.

(-MOM))；R ^13a And R ^13b Independently an amino protecting group (e.g.

(-Boc))；

R ^2a 、R ^4a 、R ⁶ 、R ⁷ 、R ^12a 、R ^13a 、A ^a 、X ^a 、L ^a 、Q ^a 、R ^2b 、R ^4b 、R ^12b 、R ^13b 、A ^b 、X ^b 、L ^b 、Q ^b The definitions of (A) and (B) are as described above.

The general formula of the compound shown in formula IIa or IIb is preferably as follows:

the compound shown in the formula IIa or IIb is preferably any one of the following compounds:

or

Indicates a chemical bond in an axial steric configuration, "-" indicates that the chiral carbon atom is in an S configuration or an R configuration.

The invention also provides a pharmaceutical composition, which comprises a substance X and one or more pharmaceutical excipients, wherein the substance X is the compound shown in the formula Ia, ib or Ic, pharmaceutically acceptable salt thereof, solvate thereof, stereoisomer thereof, tautomer thereof, prodrug thereof, metabolite thereof or isotope compound thereof.

The invention also provides the use of a substance Y in the manufacture of a medicament for the treatment or prevention of a cancer mediated by KRAS mutations; the substance Y is the compound shown in the formula Ia, ib or Ic, pharmaceutically acceptable salt, solvate, prodrug, metabolite or isotope compound thereof or the pharmaceutical composition.

In the application, the KRAS mutant protein is preferably KRAS _ G12D mutant protein.

In the application, the cancer is preferably hematological cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer and the like.

The invention also provides the use of a substance Y for the manufacture of a medicament for the treatment or prevention of cancer; the substance Y is the compound shown in the formula Ia, ib or Ic, pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, metabolite or isotope compound thereof or the pharmaceutical composition; the cancer is hematological cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer.

The present invention also provides a method of treating a cancer mediated by KRAS mutation comprising administering to a patient a therapeutically effective amount of substance Y; the substance Y is the compound shown in the formula Ia, ib or Ic, pharmaceutically acceptable salt, solvate, prodrug, metabolite or isotope compound thereof or the pharmaceutical composition.

Such as hematological cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer, and the like.

The KRAS mutein may be a KRAS _ G12D mutein.

The present invention also provides a method for the therapeutic prophylaxis or treatment of cancer, which comprises administering to a patient a therapeutically effective amount of substance Y; the substance Y is the compound shown in the formula Ia, ib or Ic, pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, metabolite or isotope compound thereof or the pharmaceutical composition; the cancer is hematological cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer.

The present disclosure also relates to a method of treating a hyperproliferative disease in a mammal comprising administering to said mammal a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof.

Ras mutations, including but not limited to K-Ras, H-Ras or N-Ras mutations that have been identified in hematological cancers or malignancies (e.g., cancers that affect blood, bone marrow and/or lymph nodes). Thus, certain embodiments relate to the administration of the disclosed compounds (e.g., in the form of a pharmaceutical composition) to a patient in need of treatment for a hematologic cancer or malignancy.

In certain particular embodiments, the present disclosure relates to methods for treating lung cancer, comprising administering to a subject in need thereof an effective amount of any of the above-described compounds (or a pharmaceutical composition comprising the same).

In the present invention, the cancer or malignant tumor includes but is not limited to leukemia and lymphoma. In certain embodiments, the leukemia is, for example, acute Lymphocytic Leukemia (ALL), acute Myelogenous Leukemia (AML), chronic Lymphocytic Leukemia (CLL), small Lymphocytic Lymphoma (SLL), chronic Myelogenous Leukemia (CML), acute monocytic leukemia (AMoL), and/or other leukemias. In certain embodiments, the lymphoma, e.g., hodgkin's lymphoma or all subtypes of non-hodgkin's lymphoma.

In certain embodiments of the invention, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung cancer or large cell lung cancer. In other embodiments, the lung cancer is small cell lung cancer. Other lung cancers include, but are not limited to, adenomas, carcinoids and undifferentiated carcinomas.

In some embodiments of the invention, the cancer, such as acute myelogenous leukemia, juvenile cancer, childhood adrenocortical carcinoma, cancers associated with AIDS (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendiceal cancer, astrocytoma, atypical malformations, basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumor, burkitt's lymphoma, carcinoid, atypical malformations, embryonic tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, cardiac tumor, chronic Lymphocytic Leukemia (CLL), chronic Myelogenous Leukemia (CML), chronic myeloproliferative disease, colon cancer, large intestine cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ carcinoma (DCIS), embryonic tumors, central nervous system cancers, endometrial cancers, ependymomas, esophageal cancers, granulomatous neuroblastoma, ewing's sarcoma, extracranial germ cell tumors, extragonadal germ cell tumors, eye cancers, bone fibrohistiocytomas, gallbladder cancers, stomach cancers, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST), germ cell tumors, gestational trophoblastic tumors, hairy cell leukemia, head and neck cancers, heart diseases, liver cancers, hodgkin's lymphoma, hypopharyngeal cancers, intraocular melanoma, islet cell tumors, pancreatic neuroendocrine tumors, kidney cancers, throat cancers, lip and oral cancers, liver cancers, lobular Carcinoma In Situ (LCIS), lung cancers, lymphomas, metastatic squamous cell cancers, occult primary, midline cancers, oral cancers, multiple endocrine tumor syndromes, multiple myeloma/plasmacytomas, mycoses, mycosis fungoides, myelodysplastic syndromes, myelodysplastic/myeloproliferative tumors, multiple myeloma, merkel cell carcinoma, malignant mesothelioma, malignant fibrous histiocytoma of bone and osteosarcoma, nasal and paranasal sinuses, nasal and sinus neuroblastoma, non-hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer, lip and oral cancer, oropharyngeal cancer, ovarian cancer, pancreatic cancer, papillomatosis, paragangliomas, paranasal and nasal cavity cancer, parathyroid cancer, penile cancer, laryngeal cancer, pleuropneumonias, primary Central Nervous System (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell cancer, retinoblastoma, rhabdomyosarcoma, salivary gland carcinoma, skin cancer, stomach (stomach) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, laryngeal and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic cell carcinoma, atypical carcinoma, pediatric sarcoma, cervical or cervical cancer. In some embodiments, the non-cancerous hyperproliferative disease, such as benign hyperplasia of the skin (e.g., psoriasis), restenosis, or prostate (e.g., benign Prostatic Hypertrophy (BPH)).

Definition of terms

The term "pharmaceutically acceptable" means that the salts, solvents, excipients, etc., are generally non-toxic, safe, and suitable for use by the patient. The "patient" is preferably a mammal, more preferably a human.

The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt as defined herein, and has all the effects of the parent compound. Pharmaceutically acceptable salts can be prepared by treating according to conventional methods with the corresponding acid in a suitable organic solvent which is an organic base.

Examples of salt formation include: for base addition salts, it is possible to prepare salts of an alkali metal (such as sodium, potassium or lithium) or of an alkaline earth metal (such as aluminum, magnesium, calcium, zinc or bismuth) by treating the compounds of the invention having suitably acidic protons in an aqueous medium with an alkali metal or alkaline earth metal hydroxide or alkoxide (such as ethoxide or methoxide) or a suitably basic organic amine (such as diethanolamine, choline or meglumine).

Alternatively, for acid addition salts, salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid; and organic acids such as acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, eurya japonica acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, glutamic acid, glycolic acid, hydroxynaphthoic acid, 2-hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, oxalic acid, pyruvic acid, malonic acid, mandelic acid, methanesulfonic acid, myfuroic acid, 2-naphthalenesulfonic acid, propionic acid, salicylic acid, succinic acid, tartaric acid, citric acid, cinnamic acid, p-toluenesulfonic acid or trimethylacetic acid.

The term "solvate" refers to a substance formed by combining a compound of the present invention with a stoichiometric or non-stoichiometric amount of a solvent. The solvent molecules in the solvate may be present in ordered or unordered arrangements. Such solvents include, but are not limited to: water, methanol, ethanol, and the like.

The term "prodrug" refers to a compound that is chemically modified to provide a drug that is inactive or less active in vitro and that is converted in vivo, enzymatically or non-enzymatically, to release the active drug and to exert its pharmacological effect.

The term "metabolite" refers to intermediate and final metabolites in metabolism.

The term "isotopic compound" means that one or more atoms in the compound can exist in its non-natural abundance. In the case of a hydrogen atom, its non-naturally abundant form refers to a form in which about 95% is deuterium.

The term "pharmaceutical excipients" may be those excipients which are widely used in the field of pharmaceutical production. The excipients are used primarily to provide a safe, stable and functional pharmaceutical composition and may also provide methods for dissolving the active ingredient at a desired rate or for promoting the effective absorption of the active ingredient after administration of the composition by a subject. The pharmaceutical excipients may be inert fillers or provide a function such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. The pharmaceutical excipients may include one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrating agents, lubricants, antiadherents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, reinforcing agents, adsorbents, buffering agents, chelating agents, preservatives, colorants, flavoring agents and sweeteners.

The pharmaceutical compositions of the present invention may be prepared according to the disclosure using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, levigating, encapsulating, entrapping or lyophilizing processes.

The pharmaceutical compositions of the invention may be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intraarterial, intramuscular). The pharmaceutical compositions of the present invention may also be in a controlled release or delayed release dosage form (e.g., liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs and solutions. Examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops or serum formulations. Examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry preparations which can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic formulations; aerosol preparation: such as nasal sprays or inhalants; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

By "treatment" is meant any treatment of a disease in a mammal, including: (1) Preventing disease, i.e., the symptoms that cause clinical disease do not develop; (2) inhibiting disease, i.e., arresting the development of clinical symptoms; (3) relieving the disease, i.e., causing regression of clinical symptoms.

By "effective amount" is meant an amount of a compound, when administered to a patient in need of treatment, that is sufficient to (i) treat the associated disease, (ii) attenuate, ameliorate, or eliminate one or more symptoms of a particular disease or condition, or (iii) delay the onset of one or more symptoms of a particular disease or condition described herein. The amount of said carbonyl heterocyclic compound of formula II or a pharmaceutically acceptable salt thereof or a pharmaceutical composition as described above that corresponds to this amount will vary depending on factors such as the particular compound, the disease condition and its severity, the characteristics of the patient in need of treatment (e.g. body weight), etc., but may nevertheless be routinely determined by one skilled in the art.

"prevention" as used herein refers to a reduction in the risk of acquiring or developing a disease or disorder.

The term "alkyl" refers to a straight or branched chain alkyl group having the indicated number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Unless a substituent is specified, an alkyl group is unsubstituted.

The term "heterocycloalkyl" means a stable 3-to 16-membered saturated cyclic group consisting of 2-11 carbon atoms and 1-5 heteroatoms selected from nitrogen, oxygen and sulfur. Unless otherwise specified in the specification, a heterocycloalkyl group can be monocyclic ("monocyclic heterocycloalkyl"), or a bicyclic, tricyclic, or higher ring system, which can include fused (fused rings), bridged (bridged rings), or spiro (spiro) ring systems (e.g., bicyclic ring systems ("bicyclic heterocycloalkyl"). Heterocycloalkyl bicyclic ring systems can include one or more heteroatoms in one or both rings, and are saturated.

The term "aryl" refers to a group having the indicated number of carbon atoms (e.g., C) ₆ ～C ₁₈ ) Cyclic, notSaturated monovalent hydrocarbon groups, which are monocyclic or polycyclic (e.g., 2 or 3), in which case the monocyclic rings share two atoms and a bond between them and (at least one ring per ring) have aromatic character, such as phenyl, naphthyl.

The term "heteroaryl" refers to an aromatic group containing a heteroatom, preferably an aromatic 5-6 membered monocyclic or 9-10 membered bicyclic ring containing 1, 2 or 3 members independently selected from nitrogen, oxygen and sulfur, for example, furyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, benzimidazolyl, indolyl, indazolyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, quinolinyl, isoquinolinyl and the like.

Used in the description structural formula of the invention

Means a chemical bond representing an axial chiral stereoconfiguration

Or

For example, compounds

Is shown as

It will be appreciated by those skilled in the art that, in accordance with common practice used in the art, the present invention describes the structural formulae used in the structural formulae of the radicals

Means that the corresponding group is linked to other fragments, groups in the compound through this site.

Used in the structural formulae of the radicals described in the invention

The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows: the compound of the invention has degradation effect on KRAS protein and has good inhibition effect on KRAS G12D mutant protein.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. Experimental procedures without specifying specific conditions in the following examples were selected in accordance with conventional procedures and conditions, or in accordance with commercial instructions.

In the present invention, the compounds and salts (or free bases) thereof finally obtained in the following examples have, if a stereoconfiguration due to axial chirality exists in these compounds, a stereoconfiguration due to axial chirality of these compounds and salts (or free bases) thereof, which is identical to that of the intermediate containing a chiral axis for the preparation of these compounds. For example compounds 1a and 1b in example 1 and their free bases: the compound 1a and its free base 1a-f (compound 1b and its free base 1 b-f) are prepared from intermediates 1-9a (intermediates 1-9 b) containing the chiral axis, so that the configuration resulting from the axis chirality in the compound 1a and its free base 1a-f (compound 1b and its free base 1 b-f) is identical to that of the intermediates 1-9a (intermediates 1-9 b). The configuration resulting from the axial chirality in other embodiments of the invention is the same as in example 1.

Free base:

in addition, the analytical parameters of intermediates 1-9a and 1-9b can be obtained by those skilled in the art by selecting and adjusting the column chromatography conditions according to the preparation parameters.

Example 1

4- ((4- (4- (3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione formate 1a;4- ((4- (4- (3- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione formate 1b.

Step 1

To a 500 ml round bottom flask were added N- (4-bromobutyl) phthalimide (10.0 g, 33.7 mmol, 1.0 equiv), acetonitrile (250.0 ml) and tert-butylpiperazine-1-carboxylic acid (7.3 g, 39.0 mmol, 1.1 equiv) in that order with stirring at 25 degrees celsius, followed by N, N-diisopropylethylamine (9.2 g, 70.9 mmol, 2.0 equiv) dropwise at 25 degrees celsius. The mixture was stirred at 80 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. And after the reaction is finished, cooling to 25 ℃, and carrying out reduced pressure concentration to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to reduced pressure rotary evaporation to remove the solvent, whereby Compound 1-1 (Red color)Oil, 13.5 g, 98% yield). MS (ESI, m/z) 388.2[ m ] +H] ⁺ 。

Step 2

Compound 1-1 (5.5 g, 13.5 mmol, 1.0 eq), ethanol (260.0 ml) and hydrazine hydrate (3.9 g, 74.2 mmol, 5.5 eq) were added sequentially to a 500 ml reaction flask at 25 ℃ with stirring under nitrogen. The resulting mixture was stirred at 80 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is cooled to room temperature, and the reaction liquid is concentrated under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 20% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 1-2 (red oil, 3.2 g, 88% yield). MS (ESI, m/z) 258.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ3.48–3.40(m,4H),2.72(t,J＝6.6Hz,2H),2.42–2.32(m,6H),1.60–1.48(m,4H),1.46(s,9H)。

Step 3

To a mixture of compound 1-2 (1.0 g, 3.5 mmol, 1.0 eq), ultra-dry N-methylpyrrolidone (10.0 ml) and tert-butyl 4- (4-aminobutyl) piperazine-1-carboxylate (1.1 g, 4.1 mmol, 1.2 eq) was added dropwise N, N-diisopropylethylamine (0.9 g, 7.2 mmol, 2.0 eq) at 25 ℃ with stirring under nitrogen. The resulting mixture was reacted at 90 ℃ for 2 hours, the course of the reaction being monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was directly purified by a reverse phase flash column (C18 column), and eluted with 5% → 60% acetonitrile/water mobile phase (0.1% formic acid) within 25 minutes; detector, UV254 nm; compound 1-3 (yellow-green oil, 374 mg, 17% yield) was obtained. MS (ESI, m/z): 514.2[M+H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ8.36(s,1H),8.24(s,1H),7.54–7.47(m,1H),7.15–7.09(m,1H),6.89(d,J＝8.5Hz,1H),6.25(s,1H),4.99–4.87(m,1H),3.66–3.55(m,4H),3.36–3.28(m,2H),2.95–2.63(m,10H),1.83–1.64(m,4H),1.46(s,9H)。

Step 4

To a solution of compound 1-3 (374.0 mg, 0.7 mmol, 1.0 eq) in dichloromethane (10.0 ml) was added trifluoroacetic acid (5.0 ml) with stirring at room temperature. The resulting mixture was stirred at 25 ℃ for an additional 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is complete. Concentration under reduced pressure removed excess reagent to give crude 1-4 (yellow solid, 300.0 mg, 88% yield). MS (ESI, m/z) 414.1[ 2 ], [ M ] +H ] ⁺ 。

Step 5

To a 500 ml round bottom flask were added 2-amino-4-bromo-5-chloro-3-fluorobenzoic acid (15.0 g, 55.9 mmol, 1.0 eq) and urea (33.6 g, 558.7 mmol, 10.0 eq) with stirring at 25 ℃. The mixture was warmed to 150 degrees celsius and stirred at that temperature for 6 hours. After the reaction is completed, the temperature is reduced to 25 ℃, 750 ml of water is added for dilution, the mixture is stirred for 30 minutes and then filtered, a filter cake is washed by water (50 ml of x 3), the solid is collected and dried under reduced pressure to obtain a compound 1-5 crude product (yellow solid, 15.5 g, purity 56%), and the compound is directly used for the next synthesis without further purification. MS (ESI, m/z): 290.9/292.9/294.8M-H] ^- ； ¹ H NMR(400MHz,DMSO-d ₆ )δ7.79(d,J＝1.8Hz,1H),6.86(s,1H),5.41(s,1H)。

Step 6

To a reaction flask were added sequentially compounds 1-5 (10.0 g, 56% pure, 19.0 mmol, 1.0 eq), N-diisopropylethylamine (15.0 ml, 81.8 mmol, 4.3 eq) and phosphorus oxychloride (150.0 ml) at 25 ℃ with stirring under nitrogen. The resulting mixture was stirred at 90 ℃ for 5 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction mixture was cooled to room temperature, and the excess reagent was removed by concentration under reduced pressure. Then 400 ml of water was added to the crude product and extracted with ethyl acetate (400 ml x 3). Combining the organic phases, washing the organic phases with 300 ml of saturated saline solution, drying the washed organic phases with anhydrous sodium sulfate, and filtering to remove the drying agent; and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 12% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 1-6 (yellow solid, 4.4 g, 70% yield). MS (ESI, m/z) 328.8/330.8/332.8[ 2 ] M + H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.21(d,J＝2.0Hz,1H)。

Step 7

To a mixture of compounds 1-6 (4.14 g, 12.5 mmol, 1.0 eq), extra dry 1, 4-dioxane (80.0 ml) and triethylamine (4.00 g, 37.5 mmol, 3 eq) was added dropwise a solution of piperazine-1-carboxylic acid tert-butyl ester (2.45 mg, 12.5 mmol, 1.0 eq) in extra dry 1, 4-dioxane (15.0 ml) with stirring at zero degrees centigrade under nitrogen protection. The mixture was reacted at 25 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, it was cooled to room temperature. The reaction was diluted with water (200 ml) and the mixture was extracted with ethyl acetate (3 × 100 ml). The organic layer was washed with saturated brine (1 × 200 ml), followed by drying over anhydrous sodium sulfate. Filtering to remove insoluble substances, and concentrating the organic phase under reduced pressure to obtain a crude product.The crude product was purified by silica gel column chromatography eluting with a 10% → 20% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 1-7 (yellow solid, 5.5 g, 84% yield). MS (ESI, m/z) 479.0/481.0/483.0[ m ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.76(d,J＝2.0Hz,1H),3.95–3.83(m,4H),3.71–3.64(m,4H),1.50(s,9H)。

Step 8

To a solution of compounds 1-7 (3.0 g, 6.0 mmol, 1.0 eq) in N-methylpyrrolidinone (30.0 ml) was added 3-buten-1-ol (1.8 g, 23.7 mmol, 4.0 eq), N-diisopropylethylamine (1.6 g, 11.9 mmol, 2.0 eq) and cesium fluoride (1.9 g, 11.9 mmol, 2.0 eq) at 25 ℃ with stirring under nitrogen. The reaction solution was stirred at 130 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the reaction was cooled to 25 ℃. The resulting mixture was extracted with dichloromethane (300 ml x 3). The combined organic layers were washed with 300 ml of water and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a 0 → 20% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compounds 1-8 (white solid, 900 mg, 29% yield). MS (ESI, m/z) 515.1/517.1/519.1M + H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.70(d,J＝2.0Hz,1H),5.97–5.87(m,1H),5.21–5.09(m,2H),4.50(t,J＝7.0Hz,2H),3.80–3.73(m,4H),3.68–3.61(m,4H),2.64–2.59(m,2H),1.49(s,9H)。

Step 9

Compounds 1-8 (620) were added at 25 c with stirring under nitrogen.0 mg, 1.2 mmol, 1.0 eq) of 1, 4-dioxane/water (5/1, 12.0 ml) was added to a solution of 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (389.7 mg, 1.4 mmol, 1.2 eq), sodium carbonate (382.2 mg, 3.6 mmol, 3.0 eq) and 1,1' -bis (diphenylphosphino) ferrocene dichloropalladium (II) (97.9 mg, 0.1 mmol, 0.1 eq). The reaction solution was stirred at 90 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the reaction solution was cooled to 25 ℃. The reaction mixture was concentrated and purified by silica gel column chromatography eluting with a 0% → 20% ethyl acetate/petroleum ether mobile phase gradient, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compounds 1 to 9 (two stereoisomer mixture, white oil, 475.0 mg, yield 67%) were obtained. MS (ESI, m/z) 579.2/581.1[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.81–7.76(m,2H),7.51–7.43(m,1H),7.37–7.31(m,2H),7.28–7.23(m,1H),7.12(d,J＝2.5Hz,1H),6.02–5.85(m,1H),5.23–5.07(m,2H),4.53(t,J＝7.0Hz,2H),3.92–3.82(m,4H),3.77–3.67(m,4H),2.68–2.58(m,2H),1.53(s,9H)。

Step 10

Chiral resolution of the compounds 1-9 (475.0 mg) obtained in step 9 by preparative chiral high performance liquid chromatography: chiral column CHIRALPAK IC,2 × 25 cm, 5 microns; a mobile phase A: n-hexane/dichloromethane =5/1 (0.1% diethylamine), mobile phase B: ethanol; flow rate: 15 ml/min; elution was performed with 50% phase B over 12 minutes with detector UV 220/235 nm to give two products. The shorter retention time (4.935 min) products were compounds 1-9a (white solid, 205.0 mg), compounds 1-9a: MS (ESI, m/z) 579.2/581.1[ M + H ] ] ⁺ (ii) a The product with longer retention time (8.413 min) was compounds 1-9b (white solid, 210.0 mg), compounds 1-9b: MS (ESI, m/z) 579.2/581.1[ 2 ], [ M ] +H] ⁺ 。

Step 11

To a solution of compounds 1-9a (180.0 mg, 0.3 mmol, 1.0 equiv) in acetone/water (4/1, 10.0. Ml) was added N-methylmorpholine oxide (72.8 mg, 0.6 mmol, 2.0 equiv) at 25 degrees celsius with stirring, followed by osmium tetroxide (39.5 mg, 0.2 mmol, 0.5 equiv) at 0 degrees celsius. The reaction solution was stirred at 25 ℃ for 3 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration was performed under reduced pressure, and the resulting mixture was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 60% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 25 minutes; detector, UV254 nm; compound 1-10a was obtained (white oil, 150.0 mg, 77% yield). MS (ESI, m/z) 613.3/615.2[ 2 ] M + H] ⁺ 。

Step 12

To a solution of compounds 1-10a (130.0 mg, 0.2 mmol, 1.0 equiv) in acetonitrile/water (4/1, 10.0 ml) was added sodium periodate (272.1 mg, 1.2 mmol, 6.0 equiv) in portions at 25 ℃ with stirring under nitrogen. The reaction solution was stirred at 25 ℃ for 0.5 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, it was extracted with dichloromethane (10 ml x 3). The combined organic layers were washed with 10 ml of saturated sodium chloride solution, and after washing, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 1-11a (oil, 123.0 mg, yield 100%). MS (ESI, m/z) 581.2/583.2[ M + H ] ] ⁺ 。

Step 13

At a temperature of 25 c, the temperature of the alloy is controlled,to a solution of compounds 1-11a (123.0 mg, 0.2 mmol, 1.0 eq) in methanol (5.0 ml) was added compounds 1-4 (105.0 mg, 0.3 mmol, 1.2 eq) with stirring, followed by glacial acetic acid (19.1 mg, 0.3 mmol, 1.5 eq) and sodium cyanoborohydride (20.0 mg, 0.3 mmol, 1.5 eq) at 25 ℃, and the reaction was stirred at 25 ℃ for 1 hour, the progress of which was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was concentrated under reduced pressure and purified by liquid chromatography on column XSelect CSH Prep C18 OBD,19X250 mm, 5 μm; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was performed with 20% → 50% B phase over 7 minutes, detector UV 220/235 nm. Compound 1-12a was obtained (yellow-green solid, 40.0 mg, 19% yield). MS (ESI, m/z) 978.4/980.4[ 2 ], [ M + H ]] ⁺ 。

Step 14

To a solution of compound 1-12a (40.0 mg, 0.04 mmol, 1 eq) in dichloromethane (5.0 ml) was added trifluoroacetic acid (2.0 ml) with stirring at room temperature. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the mixture was concentrated under reduced pressure, and the resulting mixture was purified by preparative high performance liquid chromatography on column xsselect CSH Prep C18 OBD,19 × 250 mm, 5 μm; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution with 10% → 35% B phase over 7 min, detector UV 220/235 nm gave compound 1a (yellow-green solid, 15.0 mg, 40% yield). MS (ESI, m/z) 878.4/880.4[ 2 ] M + H ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.10(s,1H),8.23–8.18(m,1H),7.97–7.93(m,1H),7.81(d,J＝8.3Hz,1H),7.60–7.54(m,1H),7.47–7.41(m,1H),7.29(d,J＝2.4Hz,1H),7.26–7.17(m,2H),7.12(d,J＝8.6Hz,1H),7.08–7.05(m,1H),7.01(d,J＝7.0Hz,1H),6.56(t,J＝6.0Hz,1H),5.08–5.01(m,1H),4.36(t,J＝6.5Hz,2H),3.86–3.77(m,4H),3.36–3.24(m,4H),3.01–2.94(m,4H),2.93–2.81(m,2H),2.64–2.51(m,2H),2.46–2.23(m,10H),2.07–1.97(m,1H),1.94–1.84(m,2H),1.62–1.42(m,4H)。

Step 11'

To a solution of compounds 1-9b (210.0 mg, 0.4 mmol, 1.0 eq) in acetone/water (4/1, 10 ml) was added N-methylmorpholine oxide (89.4 mg, 0.7 mmol, 2.0 eq) with stirring at 25 degrees celsius, followed by osmium tetroxide (46.1 mg, 0.2 mmol, 0.5 eq) at 0 degrees celsius. The reaction solution was stirred at 25 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration was carried out under reduced pressure, and the resulting mixture was directly purified by reverse phase flash chromatography (C18 column) eluting with 5% → 60% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 25 minutes; detector, UV254 nm; compounds 1-10b (white oil, 200.0 mg, 90% yield) were obtained. MS (ESI, m/z) 613.3/615.2[ 2 ] M + H] ⁺ 。

Step 12'

To a solution of compounds 1-10b (200.0 mg, 0.3 mmol, 1.0 equiv) in acetonitrile/water (4/1, 10 ml) was added sodium periodate (431.8 mg, 1.9 mmol, 6.0 equiv) in portions at 25 ℃ with stirring under nitrogen. The reaction solution was stirred at 25 ℃ for 0.5 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, it was extracted with dichloromethane (30 ml x 3). After the combination, the organic layers were washed with 30 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compounds 1-11b (oil, 189.0 mg, 100% yield). MS (ESI, m/z) 581.2/583.2[ m ] +H ] ⁺ 。

Step 13'

To a solution of compounds 1-11b (189.0 mg, 0.3 mmol, 1.0 eq) in methanol (5.0 ml) was added compounds 1-4 (146.0 mg, 0.3 mmol, 1.2 eq) at 25 deg.c with stirring, followed by glacial acetic acid (20.0 mg, 0.3 mmol, 1.2 eq) and sodium cyanoborohydride (37.0 mg, 0.6 mmol, 2.0 eq) at 25 deg.c, and the reaction was stirred at 25 deg.c for 1 hour, the course of which was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the mixture was concentrated under reduced pressure, and the resulting mixture was purified by preparative high performance liquid chromatography on column xsselect CSH Prep C18 OBD,19 × 250 mm, 5 μm; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution with 25% → 55% B phase over 7 minutes, detector UV 220/235 nm gave compounds 1-12B (yellow-green solid, 60.0 mg, 22% yield). MS (ESI, m/z) 978.4/980.4[ 2 ], [ M + H ]] ⁺ 。

Step 14'

To a solution of compound 1-12b (55.0 mg, 0.04 mmol, 1 eq) in dichloromethane (5.0 ml) was added trifluoroacetic acid (2.0 ml) with stirring at room temperature. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, concentrating under reduced pressure to remove redundant reagents, and purifying the obtained mixture by preparative high performance liquid chromatography with a chromatographic column XSelectric CSH Prep C18 OBD,19x250 mm and 5 microns; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was performed with 10% → 35% B phase over 7 minutes, detector UV 220/235 nm. Compound 1b was obtained (yellow-green solid, 20.0 mg, 36% yield). MS (Mass Spectrometry) (ESI,m/z):878.4/880.4[M+H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.10(s,1H),8.23–8.19(m,1H),7.96–7.93(m,1H),7.81(d,J＝8.3Hz,1H),7.60–7.55(m,1H),7.47–7.41(m,1H),7.29(d,J＝2.4Hz,1H),7.26–7.18(m,2H),7.12(d,J＝8.6Hz,1H),7.08–7.05(m,1H),7.01(d,J＝7.0Hz,1H),6.56(t,J＝6.0Hz,1H),5.08–5.01(m,1H),4.36(t,J＝6.5Hz,2H),3.84–3.79(m,4H),3.33–3.26(m,4H),3.01–2.94(m,4H),2.93–2.82(m,2H),2.63–2.52(m,2H),2.44–2.24(m,10H),2.06–1.98(m,1H),1.94–1.85(m,2H),1.61–1.42(m,4H)。

Example 2

4- ((5- (4- (5- ((3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) pentyl) piperazin-1-yl) pentyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione diformate 2

Step 1:

n- (5-bromopentyl) phthalimide (5.0 g, 16.8 mmol, 1.0 equivalent) and 1-tert-butoxycarbonylpiperazine (3.46 g, 1.86 mmol, 1.1 equivalent) were dissolved in 100 ml of acetonitrile with stirring at 25 degrees c, followed by addition of N, N-diisopropylethylamine (4.36 g, 33.8 mmol, 2.0 equivalents). The mixture was warmed to 80 ℃ and stirred at this temperature for 16 hours, the course of the reaction being monitored by liquid mass. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the fractions obtained were freed from the solvent by rotary evaporation under reduced pressure to give compound 2-1 (brown oil, 6.6 g, 97% yield). MS (ESI, m/z) 402.2[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.86–7.80(m,2H),7.74–7.69(m,2H),3.71–3.67(m,2H),3.45–3.42(m,4H),2.40–2.33(m,6H),1.74–1.66(m,2H),1.61–1.52(m,2H),1.46(s,9H),1.40–1.33(m,2H)。

And 2, step:

compound 2-1 (3 g, 7.1 mmol, 1.0 eq) was dissolved in 100 ml ethanol with stirring at 25 ℃, followed by the addition of hydrazine hydrate (2.57 g, 41.1 mmol, 5.5 eq). The reaction was warmed to 80 ℃ and stirred at this temperature for 16 hours, with the progress of the reaction being monitored by liquid mass. After the reaction is finished, cooling the reaction liquid to 25 ℃, then filtering the reaction liquid, washing a filter cake with 20 ml of ethanol for three times, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 20% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 2-2 (colorless oil, 1.79 g, 88% yield). MS (ESI, m/z) 272.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ3.46–3.42(m,4H),2.71–2.68(m,2H),2.38–2.32(m,6H),1.55–1.30(m,17H)。

And step 3:

2- (2, 6 oxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (700 mg, 2.5 mmol, 1.0 eq.) and compound 2-2 (756.6 g, 2.8 mmol, 1.1 eq.) were dissolved in 10 ml of N-methylpyrrolidinone with stirring at 25 deg.C under nitrogen protection, followed by the addition of N, N-diisopropylethylamine (655.1 mg, 5.1 mmol, 2.0 eq.). The reaction system was warmed to 90 ℃ and allowed to react at this temperature for 2 hours, with the progress of the reaction being monitored by liquid mass. After the reaction was completed, the reaction solution was cooled to 25 ℃. The reaction was purified by reverse phase flash chromatography (C18 column) eluting with 5 → 95% acetonitrile/water mobile phase (0.1% formic acid) over 30 minutes; detection ofUV254 nm; compound 2-3 was obtained (yellow oil, 502.3 mg, 37% yield). MS (ESI, m/z) 528.3[ 2 ], M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.29(s,1H),7.52–7.48(m,1H),7.10(d,J＝7.1Hz,1H),6.88(d,J＝8.5Hz,1H),6.24–6.22(m,1H),4.93–4.89(m,1H),3.60–3.59(m,4H),3.31–3.26(m,1H),2.91–2.59(m,15H),1.73–1.64(m,2H),1.46(s,9H)。

And 4, step 4:

compound 2-3 (200 mg, 0.36 mmol, 1.0 eq) was dissolved in 3 ml of dichloromethane with stirring at 25 ℃, the system was cooled to 0 ℃ and then 1 ml of trifluoroacetic acid was added dropwise to the mixture. After the addition, the temperature of the reaction system is raised to 25 ℃ and the reaction is carried out for 2 hours at the temperature, and the reaction process is monitored by liquid quality. After the reaction is finished, concentrating the reaction liquid under reduced pressure to obtain a crude product. The crude product 2-4 (220 mg) was obtained and used directly in the next step. MS (ESI, m/z) 428.2[ 2 ], [ M ] +H ] ⁺ 。

And 5:

5-hexenol (5.0 g, 49.9 mmol, 1.0 eq) was dissolved in 500 ml of dichloromethane with stirring at 25 ℃ under nitrogen protection, followed by addition of (1, 1-triacetoxy) -1, 1-dihydro-1, 2-phenyliodoxy-3 (1H) -one (42.4 g, 99.8 mmol, 2.0 eq). The mixture was allowed to react at 25 ℃ for 2 hours, and the progress of the reaction was monitored by thin layer chromatography. After the reaction was completed, the reaction solution was extracted with saturated sodium bicarbonate solution (500 ml × 2), the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (developer system: petroleum ether/methyl t-butyl ether = 20/1) to obtain compounds 2-5 (light yellow oil, 700 mg, yield 15%). ¹ H NMR(400MHz,DMSO-d ₆ )δ9.67(t,J＝1.5Hz,1H),5.83–5.73(m,1H),5.05–4.96(m,2H),2.45–2.41(m,2H),2.06–1.99(m,2H),1.65–1.58(m,2H)。

Step 6:

to a 500 ml reaction flask were added 3- (methylamino) propanol (10.0 g, 106.5 mmol, 1.0 eq), chloropropene (9.4 g, 123.4 mmol, 1.1 eq), N-diisopropylethylamine (29.0 g, 224.371 mmol, 2 eq) and acetonitrile (100 ml) in that order under nitrogen protection at 25 ℃. The resulting mixture was reacted at 80 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is cooled to room temperature, and the reaction liquid is concentrated under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 12% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 2-6 (yellow oil, 9.1 g, 63% yield). MS (ESI, m/z) 130.1[ m ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ6.10(s,1H),6.05–5.91(m,1H),5.42–5.36(m,2H),3.79–3.76(m,2H),3.42(d,J＝6.9,2H),2.99–2.94(m,2H),2.56(s,3H),1.94–1.86(m,2H)。

And 7:

compounds 1-7 (12.0 g, 23.7 mmol, 1.0 eq), compounds 2-6 (4.8 g, 35.6 mmol, 1.50 eq), cesium fluoride (7.2 g, 47.4 mmol, 2.0 eq) were added sequentially to a 500 ml three-neck flask under nitrogen protection at 25 ℃; n, N-diisopropylethylamine (6.1 g, 47.4 mmol, 2.0 eq) and N-methylpyrrolidinone (240 ml). The resulting mixture was reacted at 100 ℃ for 16 hours, the course of the reaction being monitored by liquid and thin layer chromatography. After the reaction is finished, the reaction solution is cooled to room temperature, filtered, and the filtrate directly passes through a reversed phase flash chromatography column (C1)8 column) and eluted with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 20 minutes; detector UV254 nm; compounds 2-7 were obtained (yellow oil, 4.5 g, 29% yield). MS (ESI, m/z) 572.1/574.1/576.1[ 2 ] M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.71(d,J＝1.96Hz,1H),6.02–5.97(m,1H),5.47–5.37(m,2H),4.53–4.50(m,2H),3.79–3.77(m,4H),3.66–3.64(m,4H),3.47(d,J＝7.04Hz,2H),3.05–3.01(m,2H),2.58(s,3H),2.28–2.21(m,2H),1.50(s,9H)。

And step 8:

to a 250 ml three-necked flask, the compounds 2-7 (4.40 g, 7.3 mmol, 1.0 eq), 4- (4, 5-tetramethyl-1, 3, 2-dioxaarene-2-yl) naphthalen-2-ol (2.70g, 9.485 mmol, 1.30 eq), potassium carbonate (2.12 g, 14.6 mmol, 2.0 eq), methanesulfonic acid (2-dicyclohexylphosphino-2 ',4',6' -tri-isopropyl-1, 1' -biphenyl) (2 ' -amino-1, 1' -biphenyl-2-yl) palladium (II) (650 mg, 0.73 mmol, 0.1 eq), 2-di-tert-butylphosphino-2 ',4',6' -triisopropylbiphenyl (366.1, 0.73 mmol, 0.1), 80 ml of 1, 4-dioxane and 20 ml of water were added in that order under nitrogen protection at 25 ℃. The resulting mixture was reacted at 60 ℃ for 3 hours, the course of the reaction being monitored by chromatography on a liquid and thin-layer chromatography. After the reaction, the reaction solution was cooled to room temperature, and the excess solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 12% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 2-8 (white solid, 2.4 g, 49% yield). MS (ESI, m/z) 636.3/638.3[ 2M + H ] ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.74–7.72(m,2H),7.43–7.39(m,1H),7.29–7.27(m,2H),7.22–7.19(m,1H),7.08(d,J＝2.4Hz,1H),5.91–5.81(m,1H),5.20–5.12(m,2H),4.49–4.42(m,2H),3.81–3.80(m,4H),3.67–3.65(m,4H),3.06(d,J＝6.64Hz,2H),2.63–2.59(m,2H),2.26(s,3H),2.09–2.02(m,2H),1.51(s,9H)。

And step 9:

chiral resolution of compounds 2-8 (2.4 g) by preparative chiral high performance liquid chromatography: chiral column CHIRALPAK IC,2 × 25 cm, 5 microns; mobile phase A: n-hexane (10 mmol/l ammonia), mobile phase B: ethanol; flow rate: 20 ml/min; elution was performed with 50% phase B over 25 minutes, detector UV 220/254 nm, to give two products. The product at the shorter retention time (5.92 min) was compound 2-8a (pale yellow oily liquid, 1.1 g, 44% recovery); the product with longer retention time (9.07 min) was compound 2-8b (pale yellow oily liquid, 0.95 g, 37% recovery).

Step 10:

to a 50 ml three-necked flask were added compounds 2-8a (1.1 g, 1.6 mmol, 1.0 eq), 1, 3-dimethyl barbituric acid (404.97 mg, 2.4 mmol, 1.5 eq), tetrakis (triphenylphosphine) palladium (99.9 mg, 0.082 mmol, 0.05 eq) and dichloromethane (20 ml) in that order under nitrogen protection at 25 ℃, and the reaction was monitored by liquid chromatography and thin layer chromatography at 25 ℃. After the reaction is finished, concentrating under reduced pressure to remove the redundant solvent to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% formic acid) over 20 minutes, detector UV254 nm to give compound 2-9a (light yellow solid, 970 mg, 94% yield). MS (ESI, m/z) 596.2/598.2[ M + H ] ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.61(s,1H),7.69–7.66(m,2H),7.35–7.31(m,1H),7.28–7.27(m,1H),7.18–7.08(m,3H),4.52–4.49(m,2H),3.82–3.64(m,8H),3.26(brs,2H),3.00–2.91(m,2H),2.50(s,3H),2.21–2.18(m,2H),1.49(s,9H)。

Step 11:

compound 2-9a (200.0 mg, 0.336 mmol, 1.0 eq), acetic acid (33.2 mg, 0.53 mmol, 1.5 eq), sodium cyanoborohydride (69.6 mg, 1.05 mmol, 3.0 eq) were dissolved in 4 ml of methanol with stirring at 25 degrees c, followed by slow dropwise addition of the compound 2-5 (54.3 mg, 0.53 mmol, 1.5 eq) and the resulting mixture was allowed to react for 2 hours with stirring at 25 degrees c, with the progress of the reaction being monitored by liquid mass. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and to obtain compound 2-10a (white solid, 190 mg, 76% yield). MS (ESI, m/z) 678.3/680.3[ M + H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.75–7.72(m,2H),7.42–7.38(t,J＝7.4Hz,1H),7.31–7.30(m,1H),7.27–7.25(m,1H),7.21–7.14(m,1H),7.15(d,J＝2.4Hz,1H),5.74–5.63(m,1H),4.97–4.90(m,2H),4.52–4.48(m,2H),3.87–3.84(m,4H),3.69–3.66(m,4H),2.99–2.94(m,2H),2.71–2.68(m,2H),2.52(s,3H),2.22–2.18(m,2H),2.06–196(m,2H),1.64–1.56(m,2H),1.51(s,9H),1.36–1.28(m,2H)。

Step 12:

compounds 2-10a (120 mg, 0.17 mmol, 1.0 equiv.) and N-methylmorpholine oxide (24.9 mg, 0.21 mmol, 1.2 equiv.) were dissolved in acetone/water (4/1, 5 mL) solution at 25 deg.C with nitrogen blanketed and stirred, followed by slow addition of potassium osmate (6.52 mg, 0.017 mmol, 0.1 equiv.) at 0 deg.C and reaction of the reaction solution at 0 deg.C with stirring for 2 hours, with the progress of the reaction monitored by liquid quality. After the completion of the reaction, the reaction mixture was purified by reverse phase flash chromatography (C18 column) at 25 minutes Elution was performed in seconds with a 5% → 95% acetonitrile/water (0.1% ammonia) mobile phase, detector UV254 nm, to give compound 2-11a (white solid, 80 mg, 63% yield). MS (ESI, m/z) 712.3/714.3[ m + H ]] ⁺ 。

Step 13:

compounds 2-11a (80.0 mg, 0.11 mmol, 1.0 equiv) were dissolved in a mixed solvent of acetonitrile/water (4/1,5 ml in volume) with stirring at 25 ℃, followed by the addition of sodium periodate (120.1 mg, 0.54 mmol, 5.0 equiv). The reaction solution reacts for 0.5 hour at 25 ℃ under stirring, and the reaction process is monitored by liquid quality. After the reaction, the reaction solution was diluted with 20 ml of water, extracted with dichloromethane (15 ml × 3), the organic phases were combined, the combined organic phases were washed with 50 ml of saturated brine, the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude product 2-12a (80 mg, yield 88%) which was used in the next reaction. MS (ESI, m/z) 680.3/682.3[ m + H ]] ⁺ 。

Step 14:

compound 2-4 (64.8 mg) and sodium cyanoborohydride (18.7 mg, 0.28 mmol, 3.0 equiv.) were dissolved in 3 ml of methanol with stirring at 25 ℃ under nitrogen, followed by addition of compound 2-12a (80.0 mg, 0.09 mmol, 1.0 equiv.). The mixture was reacted for 1 hour at 25 ℃ with stirring, and the progress of the reaction was monitored by liquid mass. After the reaction was completed, the reaction solution was purified by preparative silica gel thin layer chromatography (developing solvent system: dichloromethane/methanol = 7/1) to obtain compound 2-13a (yellow oil, 90.0 mg, yield 83%). MS (ESI, m/z) 1091.5/1093.5[ m ] +H ] ⁺ 。

Step 15:

compound 2-13a (90.0 mg, 0.082 mmol, 1.00 eq) was dissolved in 3 ml of dichloromethane with stirring at 25 degrees celsius, the reaction was brought to 0 degrees celsius, and 1 ml of trifluoroacetic acid was added dropwise. The reaction solution reacts for 1 hour under the condition of stirring at 25 ℃, and the reaction process is monitored by liquid quality. After the reaction is finished, concentrating the reaction solution under reduced pressure to obtain a crude product, and purifying the obtained crude product by preparative high performance liquid chromatography under the following preparation conditions: reversed phase Column XBridge Shield RP18 OBD Column,19 × 150 mm, 5 μm; a mobile phase A: water (0.1% formic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; eluting with 8-35% mobile phase B for 7 min; detector UV254/220 nm; compound 2 was obtained (yellow solid, 30 mg, 33.7% yield). MS (ESI, m/z) 991.5/993.5[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.50(s,2H),8.03(d,J＝1.6Hz,1H),7.79–7.76(m,1H),7.59–7.54(m,1H),7.46–7.41(m,1H),7.29(d,J＝2.4Hz,1H),7.26–7.18(m,2H),7.08–7.05(m,3H),5.09–5.03(m,1H),4.62–4.58(m,2H),4.10–4.06(m,4H),3.39–3.37(m,5H),3.29(s,4H),3.13–3.08(m,2H),2.94–2.57(m,17H),2.32–2.23(m,2H),2.18–2.06(m,1H),1.82–1.36(m,12H)； ¹⁹ F NMR(282MHz,CD ₃ OD)δ-123.15。

Example 3

4- ((5- (4- (5- ((3- ((S or R) -4- ((1r, 5s) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) pentyl) piperazin-1-yl) pentyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione diformate 3a;4- ((5- (4- (5- ((3- ((R or S) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) pentyl) piperazin-1-yl) pentyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione bistrifluoroacetate salt 3b

Step 1

To a 250 ml single neck flask were added 3- (methylamino) -1-propanol (3.0 g, 33.7 mmol, 1.0 eq) and acetonitrile (30.0 ml) with stirring at 25 ℃, followed by 6-chloro-1-hexene (4.0 g, 33.7 mmol, 1.0 eq) and potassium carbonate (9.3 g, 67.3 mmol, 2.0 eq). The resulting mixture was stirred at 70 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, cooling the mixture to 25 ℃, filtering, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 12% methanol (2.5% methanolic ammonia) in dichloromethane and the resulting fraction was stripped of solvent by rotary evaporation under reduced pressure to give compound 3-1 (yellow oil, 3.3 g, 54% yield). MS (ESI, m/z) 172.2[ 2 ], [ M ] +H] ⁺ ； ¹ HNMR(300MHz,CDCl ₃ )δ5.89–5.74(m,1H),5.07–4.93(m,2H),3.86–3.79(m,2H),2.65–2.59(m,2H),2.42–2.36(m,2H),2.26(s,3H),2.14–2.03(m,2H),1.77–1.67(m,2H),1.59–1.33(m,4H)。

Step 2

To a solution of compounds 1-6 (6.00 g, 17.3 mmol, 1.0 eq) in ultra dry dichloromethane (70.0 ml) was added N, N-diisopropylethylamine (7.04 g, 51.7 mmol, 3 eq) at 0 deg.C with stirring under nitrogen, followed by dropwise addition of 3, 8-diazabicyclo [3.2.1 ℃ ]]Octane-8-carboxylic acid tert-butyl ester (3.86 g, 17.3 mmol, 1.0 equiv.) in ultra dry dichloromethane (30.0 ml). The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, it was cooled to room temperature. The mixture is concentrated under reduced pressure to obtain And (4) obtaining a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 50% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 3-2 (yellow solid, 8.1 g, 88% yield). MS (ESI, m/z) 505.0/507.1/509.1[ 2 ] M + H +] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.75(d,J＝2.0Hz,1H),4.52–4.29(m,4H),3.79–3.50(m,2H),2.04–1.89(m,2H),1.80–1.66(m,2H),1.52(s,9H)。

Step 3

To a solution of compound 3-2 (2.0 g, 3.8 mmol, 1.00 eq) in N-methylpyrrolidinone (20.0 ml) was added, under nitrogen protection at 25 degrees celsius, 3-1 (1.4 g, 7.5 mmol, 2.0 eq), cesium fluoride (1.2 g, 7.5 mmol, 2.0 eq) and N-diisopropylethylamine (1.5 g, 11.2 mmol, 3.0 eq), in that order. The resulting mixture was reacted at 120 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was cooled to 25 ℃. The reaction was directly purified by reverse phase chromatography (C18 column) eluting with 5% → 95% methanol/water (0.1% ammonium bicarbonate) mobile phase over 25 minutes, detector UV254/220 nm to give compound 3-3 (red oil, 1.4 g, 52% yield). MS (ESI, m/z) 640.2/642.2/644.2[ 2 ] M + H +] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.71(d,J＝2.0Hz,1H),5.88–5.72(m,1H),5.08–4.87(m,2H),4.56–4.46(m,2H),4.44–4.26(m,4H),3.68–3.48(m,2H),2.76–2.64(m,2H),2.56–2.44(m,2H),2.37(s,3H),2.18–2.03(m,4H),2.02–1.92(m,2H),1.84–1.73(m,2H),1.66–1.56(m,2H),1.53(s,9H),1.48–1.36(m,2H)。

Step 4

Under the protection of nitrogen at 25 ℃, the volume is increased to 100 milliliters To a single-neck flask were added the compounds 3-3 (1.4 g, 2.2 mmol, 1.0 eq), 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (0.8 g, 2.8 mmol, 1.3 eq), chloro (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl ] in that order]Palladium (II) (166.4 mg, 0.2 mmol, 0.1 eq), potassium phosphate (0.9 g, 4.1 mmol, 2.0 eq), and tetrahydrofuran/water (10/1, 15.0 ml). The resulting mixture was stirred at 60 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the mixture is cooled to 25 ℃, and a crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 3-4 (red oil, 1.2 g, 78% yield). MS (ESI, m/z): 704.3/706.3[ M + H ]] ⁺ ； ¹ HNMR(400MHz,CDCl ₃ )δ7.75–7.67(m,2H),7.41–7.35(m,1H),7.32(d,J＝2.4Hz,1H),7.26–7.23(m,1H),7.20–7.14(m,2H),5.73–5.62(m,1H),4.97–4.88(m,2H),4.52–4.41(m,4H),4.40–4.31(m,2H),3.75–3.47(m,2H),2.97–2.87(m,2H),2.68–2.57(m,2H),2.46(s,3H),2.26–2.15(m,2H),2.04–1.95(m,4H),1.86–1.75(m,2H),1.66–1.58(m,2H),1.53(s,9H),1.34–1.27(m,2H)。

Step 5

Chiral resolution is carried out on the compound 3-4 (1.2 g) obtained in the step 4, and the resolution conditions are as follows: chiral column NB-Lux 5 μm i-Cellulose-5,2.12x25 cm, 5 μm); a mobile phase A: n-hexane/dichloromethane (5/1) (0.5% 2 mol/l ammonia methanol), mobile phase B ethanol; flow rate: 20 ml/min; column temperature: 35 ℃ is carried out; elution with 0% → 20% mobile phase B for 20 minutes; detector UV220/254 nm; two products were obtained. Short Retention Time (RT) ₁ Compound 3-4a (white solid, 520.0 mg, recovery 43%) MS (ESI, m/z) 704.3/706.3[ 2 ] M + H +] ⁺ (ii) a Longer Retention Time (RT) ₂ Compound of =10.5 min) is 3-4b (white solid, 480.0 mg, recovery 40%), MS (ESI, m/z): 704.3/706.3[ m + h ] +] ⁺ 。

Step 6

3-4a (114.0 mg, 0.14 mmol, 1.0 equiv.) was dissolved in the mixed solution of acetone/water (4/1, 2.0 mL) with stirring at 25 deg.C, followed by addition of potassium osmate (5.2 mg, 0.02 mmol, 0.1 equiv.) and then N-methylmorpholine oxide (25.0 mg, 0.2 mmol, 1.5 equiv.) with stirring at 0 deg.C. The reaction solution was reacted at 0 ℃ for 2 hours, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was returned to 25 ℃. The reaction solution was directly purified by a reverse phase chromatography column (C18 column) and eluted with a 5% → 95% methanol/water (0.1% ammonia) mobile phase in 25 minutes; detector UV254/220 nm; compound 3-5a was obtained (brown oil, 92.0 mg, 88% yield). MS (ESI, m/z) 738.4/740.4[ m + H ], [] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.75–7.67(m,2H),7.41–7.34(m,1H),7.29–7.27(m,1H),7.26–7.24(m,1H),7.21–7.15(m,1H),7.10–7.06(m,1H),4.73–4.65(m,1H),4.52–4.28(m,6H),3.66–3.57(m,2H),3.55–3.50(m,1H),3.38–3.30(m,1H),2.62–2.54(m,2H),2.40–2.36(m,2H),2.22(s,3H),2.04–1.77(m,6H),1.55–1.30(m,15H)。

Step 7

Compound 3-5a (92.0 mg, 0.14 mmol, 1.0 eq) was dissolved in acetonitrile/water (4/1, 4.0 ml) with stirring at 25 ℃, followed by the addition of sodium periodate (181.7 mg, 0.9 mmol, 6.0 eq). The resulting mixture was reacted at 25 ℃ for 0.5 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction is carried out The reaction solution was poured into 10 ml of water and then extracted with dichloromethane (10 ml x 3). The combined organic phases were washed with a further 20 ml of saturated brine. After washing, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product 3-6a (105.1 mg) which was used directly in the next step. MS (ESI, m/z) 706.4/708.4[ m + H ]] ⁺ 。

Step 8

To a 50 ml single-neck flask were added sequentially compounds 2-4 (80.6 mg, 0.15 mmol, 1.0 eq), sodium cyanoborohydride (29.5 mg, 0.5 mmol, 3.0 eq) and methanol (3.0 ml) with stirring at 25 ℃. Then, compound 3-6a (105.1 mg, 0.2 mmol, 1.0 eq) was dissolved in 2.0 ml of methanol and slowly added dropwise to the reaction solution. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Purifying the crude product by using a high performance liquid phase, wherein the preparation conditions are as follows: reversed phase Column XSelect CSH Prep C18 OBD Column,19X250 mm, 5 micron; a mobile phase A: water (0.1% formic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; elution with 17% → 43% mobile phase B for 7 minutes; detector UV254/220 nm; product 3-7a was obtained (yellow oil, 110.0 mg, 66% yield). MS (ESI, m/z) 1117.5/1119.5[ m + H ] ] ⁺ 。

Step 9

Compound 3-7a (110.0 mg, 0.1 mmol, 1.0 eq) was dissolved in dichloromethane (2.0 ml) with stirring at 25 degrees celsius and trifluoroacetic acid (1.0 ml) was added with stirring at 0 degrees celsius. After the reaction solution is added, the temperature is restored to 25 ℃, the reaction solution is stirred and reacted for 1 hour at the temperature of 25 ℃, and the reaction process is monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentratedAnd (4) condensing to obtain a crude product. Purifying the crude product by using a high performance liquid phase, wherein the preparation conditions are as follows: reversed phase Column XSelect CSH Prep C18 OBD Column,19X250 mm, 5 micron; mobile phase A: water (0.1% formic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; elution with 5% → 16% mobile phase B for 7 minutes; detector UV254/220 nm; product 3a was obtained (yellow solid, 27 mg, 26% yield). MS (ESI, m/z) 1017.5/1019.5[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.52(s,2H),8.02–7.98(m,1H),7.81–7.73(m,1H),7.60–7.52(m,1H),7.47–7.39(m,1H),7.29(d,J＝2.4Hz,1H),7.28–7.18(m,2H),7.09–7.03(m,3H),5.10–5.02(m,1H),4.70–4.54(m,4H),4.06–3.96(m,2H),3.82(d,J＝13.4Hz,2H),3.40–3.35(m,2H),3.32–3.25(m,2H),3.13–3.04(m,2H),2.94–2.52(m,18H),2.33–2.20(m,2H),2.16–1.98(m,5H),1.84–1.55(m,8H),1.54–1.36(m,4H)； ¹⁹ F NMR(282MHz,CD ₃ OD)δ-123.18。

Step 6'

Compounds 3-4b (120.0 mg, 0.17 mmol, 1.0 equiv.) are dissolved in acetone/water (4/1, 2.0 mL) with stirring at 25 deg.C, followed by the addition of potassium osmate (6.3 mg, 0.02 mmol, 0.1 equiv.) and then N-methylmorpholine oxide (30.0 mg, 0.3 mmol, 1.5 equiv.) with stirring at 0 deg.C. The reaction solution was reacted at 0 ℃ for 2 hours, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was returned to 25 ℃. The reaction solution was directly purified by a reverse phase chromatography column (C18 column), and eluted with a mobile phase of 5% → 95% methanol/water (0.1% ammonia) in 25 minutes; detector UV254/220 nm; compound 3-5b was obtained (brown oil, 100.0 mg, 80% yield). MS (ESI, m/z) 738.4/740.4[ m + H ], [ ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.74–7.68(m,2H),7.41–7.36(m,1H),7.29–7.24(m,2H),7.21–7.15(m,1H),7.10–7.07(m,1H),4.54–4.38(m,4H),4.36–4.27(m,2H),3.71–3.56(m,2H),3.55–3.50(m,1H),3.49(s,1H),3.38–3.30(m,1H),2.63–2.52(m,2H),2.43–2.33(m,2H),2.23(s,3H),2.08–1.90(m,4H),1.87–1.74(m,2H),1.52(s,9H),1.50–1.42(m,2H),1.40–1.28(m,4H)。

Step 7'

Compound 3-5b (100.0 mg, 0.14 mmol, 1.0 eq) was dissolved in acetonitrile/water (4, 1,4.0 ml) with stirring at 25 degrees celsius, and sodium periodate (165.1 mg, 0.8 mmol, 6.0 eq) was added. The reaction solution was reacted at 25 ℃ for 0.5 hour, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was poured into 10 ml of water, and then extracted with dichloromethane (10 ml × 3). The combined organic phases were washed with a further 20 ml of saturated brine. After washing, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude 3-6b (95.0 mg) which was used directly in the next step. MS (ESI, m/z) 706.4/708.4[ m + H ]] ⁺ 。

Step 8'

To a 50 ml single neck flask were added sequentially compounds 2-4 (72.8 mg, 0.13 mmol, 1.00 eq), sodium cyanoborohydride (25.4 mg, 0.4 mmol, 3.0 eq) and methanol (3.0 ml) with stirring at 25 ℃. Then, compound 3-6b (95.0 mg, 0.1 mmol, 1.0 eq) was dissolved in 2.0 ml of methanol and slowly added dropwise to the reaction solution. The mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. The crude product was purified by high performance liquid chromatography, conditions for preparation, reversed phase Column xsselect CSH Prep C18 OBD Column,19 × 250 mm, 5 μm, mobile phase a: water (0.1% formic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; elution with 17% → 43% mobile phase B for 7 minutes; detector UV254/220 Nano; product 3-7b was obtained (yellow oil, 120.0 mg, 76% yield). MS (ESI, m/z) 1117.5/1119.5[ 2 ], [ M + H ]] ⁺ 。

Step 9'

Compounds 3-7b (120.0 mg, 0.1 mmol, 1.0 eq) were dissolved in dichloromethane (2.0 ml) with stirring at 25 ℃ and trifluoroacetic acid (1.0 ml) was added with stirring at 0 ℃. After the reaction solution is added, the temperature is restored to 25 ℃, the reaction solution is stirred and reacted for 1 hour at the temperature of 25 ℃, and the reaction process is monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Purifying the crude product by using a high performance liquid phase, wherein the preparation conditions are as follows: reversed phase Column XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; elution with 5% → 16% mobile phase B for 7 minutes; detector UV 254/220 nm; product 3b was obtained (yellow solid, 30.0 mg, 23% yield). MS (ESI, m/z) 1017.5/1019.5[ m ] +H] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.03–7.98(m,1H),7.81–7.75(m,1H),7.62–7.52(m,1H),7.47–7.39(m,1H),7.29(d,J＝2.4Hz,1H),7.28–7.17(m,2H),7.10–7.03(m,3H),5.12–5.02(m,1H),4.74–4.54(m,4H),4.21–4.15(m,2H),3.94–3.83(m,2H),3.42–3.35(m,4H),3.22–3.13(m,2H),3.02–2.58(m,18H),2.36–2.23(m,2H),2.18–2.05(m,5H),1.86–1.56(m,8H),1.55–1.36(m,4H)； ¹⁹ F NMR(282MHz,CD ₃ OD)δ-123.07。

Example 4

4- ((4- (4- (4- ((3- ((4- ((1r, 5s) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- ((S or R) -3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione tetratrifluoroacetate 4a;4- ((4- (4- (4- ((3- ((4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- ((R or S) -3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione tetratrifluoroacetate 4b.

Step 1

To the reaction flask were added 3- (methylamino) -1-propanol (2.50 g, 26.6 mmol, 1.00 eq), acetic acid (2.40 g, 40.0 mmol, 1.50 eq), sodium cyanoborohydride (5.02 g, 79.9 mmol, 3.00 eq) and methanol (30 ml) in that order with stirring at 25 ℃. 4-pentenal (2.60 g, 29.4 mmol, 1.10 equiv) was then added dropwise thereto. The resulting mixture was stirred at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 11% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 4-1 (pale yellow oil, 4.4 g, 99% yield). MS (ESI, m/z) 158.2[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ5.84–5.72(m,1H),5.15–5.05(m,2H),3.84(t,J＝5.5Hz,2H),3.20(t,J＝6.8Hz,2H),3.05–2.97(m,2H),2.80(s,3H),2.25–2.14(m,2H),2.04–1.92(m,2H),1.91–1.78(m,2H)。

Step 2

To a solution of compound 3-2 (2.0 g, 3.8 mmol, 1.0 eq) in N-methylpyrrolidone (20.0 ml) was added compound 4-1 (1.2 g, 7.5 mmol, 2.0 eq), cesium fluoride (1.2 g, 7.5 mmol, 2.0 eq) and N-diisopropylethylamine (1.5 g, 11.3 mmol) in that order under nitrogen protection at 25 degrees celsius3.0 equivalents). The reaction solution was reacted at 120 ℃ for 2 hours, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was cooled to 25 ℃. Filtering, and purifying the filtrate directly through a reverse phase chromatographic column (C18 column) by eluting with 5% → 95% methanol/water (0.1% ammonium bicarbonate) mobile phase within 25 minutes; detector UV254/220 nm gave compound 4-2 (orange oil, 1.0 g, 43% yield). MS (ESI, m/z) 626.2/628.1/630.1M + H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.69(d,J＝2.0Hz,1H),5.86–5.72(m,1H),5.08–4.90(m,2H),4.48(t,J＝6.4Hz,2H),4.45–4.25(m,4H),3.70–3.45(m,2H),2.69(t,J＝7.2Hz,2H),2.50(t,J＝7.6Hz,2H),2.36(s,3H),2.17–2.03(m,4H),2.00–1.92(m,2H),1.82–1.74(m,2H),1.70–1.60(m,2H),1.52(s,9H)。

Step 3

The compounds 4-2 (1.0 g, 1.5 mmol, 1.0 eq.) and 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (646.3 mg, 2.4 mmol, 1.5 eq.), chlorine (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl ] were added sequentially to a 100 ml single neck flask under nitrogen protection at 25 deg.C]Palladium (II) (125.5 mg, 0.2 mmol, 0.1 eq), potassium phosphate (677.1 mg, 3.0 mmol, 2.0 eq) and tetrahydrofuran/water (10/1, 11.0 ml) mixed solvent. The resulting mixture was stirred at 60 ℃ for two hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the mixture is cooled to 25 ℃, and a crude product is obtained by decompression and concentration. The crude product was purified by column chromatography on silica gel eluting with a gradient of 0% → 10% methanol/dichloromethane and the fractions were freed from the solvent by rotary evaporation under reduced pressure to give compound 4-3 (red oil, 800.0 mg, 77% yield). MS (ESI, m/z) 690.3/692.3[ m ] +H] ⁺ ； ¹ HNMR(400MHz,CDCl ₃ )δ7.78–7.70(m,2H),7.44–7.37(m,1H),7.32–7.26(m,2H),7.23–7.17(m,1H),7.13–7.10(m,1H),5.78–5.65(m,1H),5.01–4.88(m,2H),4.55–4.25(m,6H),3.76–3.45(m,2H),2.86–2.72(m,2H),2.62–2.48(m,2H),2.38(s,3H),2.20–2.08(m,2H),2.06–1.92(m,4H),1.86–1.76(m,2H),1.73–1.60(m,2H),1.53(s,9H)。

Step 4

Carrying out chiral resolution on the compound 4-3 (800 mg) obtained in the step 3 under the resolution conditions that: chiral column CHIRALPAK IA,2 × 25 cm, 5 μm; mobile phase A: n-hexane (10 mmol/l ammonia methanol), mobile phase B isopropanol; flow rate: 20 ml/min; elution with 30% mobile phase B for 22 min; detector UV220/210 nm; two products were obtained. Short Retention Time (RT) ₁ Compound of =10 min) is 4-3a (red solid, 330.0 mg, recovery 41%), MS (ESI, m/z): 690.3/692.3[ M + H ])] ⁺ (ii) a Longer Retention Time (RT) ₂ Compound of =16 min) is 4-3b (red solid, 230.0 mg, 29% recovery), MS (ESI, m/z): 690.3/692.3[ M + H ])] ⁺ 。

Step 5

Compound 4-3a (200.0 mg, 0.3 mmol, 1.0 equiv.) is dissolved in acetone/water (4/1, 5.0 mL) with stirring at 25 deg.C, followed by the addition of potassium osmate (10.7 mg, 0.03 mmol, 0.1 equiv.) and then N-methylmorpholine oxide (50.9 mg, 0.4 mmol, 1.5 equiv.) with stirring at 0 deg.C. The resulting mixture was reacted at 0 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was returned to 25 ℃. The reaction solution was directly purified by a reverse phase chromatography column (C18 column) and eluted with a 5% → 95% methanol/water (0.1% ammonia) mobile phase in 25 minutes; detector UV254/220 nm; to obtain compound 4-4a (Red solid)180.0 mg, 86% yield). MS (ESI, m/z) 724.3/726.3[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.76–7.68(m,2H),7.42–7.36(m,1H),7.31–7.27(m,2H),7.22–7.16(m,1H),7.15–7.11(m,1H),4.56–4.26(m,6H),3.76–3.32(m,6H),2.74–2.62(m,1H),2.60–2.50(m,1H),2.48–2.34(m,2H),2.25(s,3H),2.10–1.93(m,4H),1.89–1.76(m,2H),1.72–1.60(m,3H),1.52(s,9H)。

Step 6

Compound 4-4a (75.0 mg, 0.1 mmol, 1.0 eq) was dissolved in acetonitrile/water (4, 2.0 ml) with stirring at 25 degrees celsius, and sodium periodate (132.9 mg, 0.6 mmol, 6.0 eq) was added. The resulting mixture was reacted at 25 ℃ for 0.5 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was poured into 10 ml of water, and then extracted with dichloromethane (15 ml × 3). The combined organic phases were washed with 15 ml of saturated brine. After the organic phase was dried over anhydrous sodium sulfate, it was filtered, and the filtrate was concentrated under reduced pressure to give crude 4-5a (60.0 mg) which was used directly in the next step. MS (ESI, m/z) 692.9/694.9[ 2 ] M + H ] ⁺ 。

Step 7

To a 50 ml single-neck flask were added sequentially compounds 1-4 (35.8 mg, 0.1 mmol, 1.0 eq), sodium cyanoborohydride (16.3 mg, 0.2 mmol, 3.0 eq) and methanol (3.0 ml) with stirring at 25 ℃. Then, compound 4-5a (60.0 mg, 0.1 mmol, 1.00 eq) was dissolved in 2.0 ml of methanol and slowly added dropwise to the reaction solution. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. The crude product obtained is passed through silica gelPurification by column chromatography was carried out with a mobile phase eluted with a gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, giving compound 4-6a (yellow solid, 62.0 mg, 66% yield). MS (ESI, m/z) 1089.5/1090.5[ M ] +H] ⁺ 。

Step 8

Compound 4-6a (62.0 mg, 0.1 mmol, 1.0 eq) was dissolved in dichloromethane (3.0 ml) with stirring at 25 ℃ and trifluoroacetic acid (1.0 ml) was added with stirring at 0 ℃. After the reaction solution is added, the temperature is restored to 25 ℃, the reaction solution is stirred and reacted for 1 hour at the temperature of 25 ℃, and the reaction process is monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. The crude product is purified by high performance liquid phase, and the preparation conditions are as follows: high-pressure reversed-phase Column XSelect CSH Prep C18 OBD Column,19x250 mm, 5 micron; mobile phase A: water (0.1% formic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; eluting with 25% -54% mobile phase B for 7 min; detector UV254/220 nm; product 4a was obtained (yellow solid, 38.0 mg, 46% yield). MS (ESI, m/z) 990.4/992.4[ m ] +H ] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.00(s,1H),7.78–7.73(m,1H),7.68–7.52(m,1H),7.45–7.38(m,1H),7.27(d,J＝2.4Hz,1H),7.24–7.16(m,2H),7.08–7.01(m,3H),5.08–5.01(m,1H),4.76–4.68(m,2H),4.64–4.54(m,2H),4.25(s,2H),3.96–3.86(m,2H),3.50–3.34(m,4H),3.28–3.10(m,6H),3.09–2.97(m,5H),2.96–2.63(m,9H),2.35–2.24(m,2H),2.19–2.04(m,5H),1.88–1.62(m,8H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-77.14,-123.09。

Step 5'

Compounds 4-3b (200.0 mg, 0.3 mmol, 1.0 equiv.) were dissolved with stirring at 25 deg.CTo acetone/water (4/1, 5.0 mL) was added potassium osmate (10.7 mg, 0.03 mmol, 0.1 equiv) followed by N-methylmorpholine oxide (50.9 mg, 0.4 mmol, 1.5 equiv) with stirring at 0 deg.C. The resulting mixture was reacted at 0 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was returned to 25 ℃. The reaction solution was directly purified by a reverse phase chromatography column (C18 column) and eluted with 5% → 95% methanol/water mobile phase (0.1% ammonia) within 25 minutes; detector UV254/220 nm; compound 4-4b was obtained (red solid, 170.0 mg, 81% yield). MS (ESI, m/z): 724.3/726.3[ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.75–7.69(m,2H),7.42–7.35(m,1H),7.31–7.27(m,2H),7.22–7.15(m,1H),7.14–7.11(m,1H),4.54–4.28(m,6H),3.74–3.32(m,6H),2.74–2.61(m,1H),2.59–2.47(m,1H),2.46–2.32(m,2H),2.26–2.20(m,3H),2.08–1.92(m,4H),1.88–1.74(m,2H),1.70–1.59(m,3H),1.52(s,9H)。

Step 6'

Compound 4-4b (80.0 mg, 0.1 mmol, 1.0 eq) was dissolved in acetonitrile/water (4, 1,2.0 ml) with stirring at 25 degrees celsius, and sodium periodate (141.8 mg, 0.6 mmol, 6.0 eq) was added. The resulting mixture was reacted at 25 ℃ for 0.5 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was poured into 10 ml of water, and then extracted with dichloromethane (15 ml × 3). The combined organic phases were washed with 20 ml of saturated brine, the washed organic phases were dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give crude 4-5b (70.0 mg) which was used directly in the next step. MS (ESI, m/z) 692.9/694.9[ M ] +H ] ⁺ 。

Step 7'

To a 50 ml single-neck flask were added sequentially compounds 1-4 (42.6 mg, 0.1 mmol, 1.0 eq), sodium cyanoborohydride (19.3 mg, 0.2 mmol, 3.0 eq) and methanol (3.0 ml) with stirring at 25 ℃. Then, compound 4-5b (70.0 mg, 0.1 mmol, 1.0 eq) was dissolved in 2.0 ml of methanol and slowly added dropwise to the reaction solution. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary distilled under reduced pressure to remove the solvent and afford compounds 4-6b (yellow solid, 73.0 mg, 65% yield). MS (ESI, m/z) 1089.5/1090.5[ M ] +H] ⁺ 。

Step 8'

Compound 4-6b (73.0 mg, 0.1 mmol, 1.0 eq) was dissolved in dichloromethane (3.0 ml) with stirring at 25 ℃ and trifluoroacetic acid (1.0 ml) was added with stirring at 0 ℃. After the addition reaction was completed, the temperature was returned to 25 ℃ and the reaction was stirred at 25 ℃ for 1 hour, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Purifying the crude product by using a high performance liquid phase, wherein the preparation conditions are as follows: high-pressure reversed-phase Column XSelect CSH Prep C18 OBD Column,19x250 mm, 5 micron; a mobile phase A: water (0.1% formic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; elution with 25% → 54% mobile phase B for 7 minutes; detector UV 254/220 nm; product 4b was obtained (yellow solid, 52.0 mg, 78% yield). MS (ESI, m/z) 990.4/992.4[ m + H ], [ solution ] M + ] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.02–7.98(m,1H),7.78–7.72(m,1H),7.59–7.52(m,1H),7.45–7.38(m,1H),7.27(d,J＝2.4Hz,1H),7.24–7.16(m,2H),7.09–7.01(m,3H),5.09–5.01(m,1H),4.76–4.68(m,2H),4.64–4.54(m,2H),4.25(s,2H),3.95–3.86(m,2H),3.51–3.34(m,4H),3.28–3.17(m,5H),3.17–2.97(m,6H),2.92(s,3H),2.91–2.62(m,6H),2.34–2.24(m,2H),2.20–2.04(m,5H),1.88–1.64(m,8H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-77.19,-123.09。

Example 5

4- ((4- (4- ((3- ((4- ((1s, 4s) -2, 5-diazabicyclo [2.2.2] octan-2-yl) -6-chloro-8-fluoro-7- ((S or R) -3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione formate 5a;4- ((4- (4- ((3- ((4- ((1s, 4s) -2, 5-diazabicyclo [2.2.2] octan-2-yl) -6-chloro-8-fluoro-7- ((R or S) -3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione diformate 5b

Step 1:

7-bromo-2, 4, 6-trichloro-8-fluoroquinazoline (780 mg, 2.2 mmol, 1.0 eq.) and triethylamine (716.8 mg, 7.1 mmol, 3.0 eq.) were dissolved in 15 ml of 1, 4-dioxane with stirring at 25 deg.C under nitrogen protection, the system was cooled to 0 deg.C, and (1S, 4S) -2, 5-diazabicyclo [2.2.2] was added]Octane-2-carboxylic acid tert-butyl ester (501.2 mg, 2.36 mmol, 1.0 eq). The mixture was reacted for 2 hours at 25 ℃ with stirring, the course of the reaction being monitored by liquid mass. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. The crude product obtained was purified by silica gel column chromatography (mobile phase eluted with a gradient of 0% → 20% ethyl acetate/dichloromethane) and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 5-1 (yellow solid, 1.16 g, 97% yield). MS (ESI, m/z) 505.1/507.1/509.1M + H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.98–7.88(m,1H),5.17(s,1H),4.53–4.20(m,2H),4.05–3.92(m,1H),3.83–3.75(m,1H),3.67–3.52(m,1H),2.36–2.08(m,2H),2.02–1.74(m,2H),1.48(s,9H)。

Step 2:

under the condition of nitrogen protection and stirring at 25 ℃, the compound 5-1 (1.1 g, 2.06 mmol, 1.0 equivalent) and 3- [ methyl (pent-4-en-1-yl) amino]Propanol (683.5 mg, 4.1 mmol, 2.0 equiv.) was dissolved in 10 ml of N-methylpyrrolidinone, followed by the addition of N, N-diisopropylethylamine (842.6 mg, 6.2 mmol, 3.0 equiv.) and cesium fluoride (660.2 mg, 4.1 mmol, 2.0 equiv.). After the addition, the temperature of the system is raised to 100 ℃ and the reaction is carried out for 4 hours at the temperature, and the reaction process is monitored by liquid quality. After the reaction, the reaction solution was cooled to 25 ℃, purified by a reverse phase flash column (C18 column), and eluted with a methanol/water (0.1 ammonia) mobile phase of 10% → 95% in 20 minutes; detector UV254 nm; compound 5-2 was obtained (white solid, 670 mg, 49% yield). MS (ESI, m/z) 626.2/628.2/630.2M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.87–7.83(m,1H),5.83–5.73(m,1H),5.08–4.95(m,3H),4.50–4.47(m,3H),4.31–4.21(m,1H),4.00–3.92(m,1H),3.81–3.77(m,1H),3.63–3.53(m,1H),2.79–2.76(m,2H),2.60–2.56(m,2H),2.42(s,3H),2.31–2.24(m,1H),2.17–2.06(m,5H),2.97–1.88(m,1H),1.86–1.76(d,J＝10.5Hz,1H),1.74–1.66(m,2H),1.47(s,9H)。

And 3, step 3:

to a 40 ml reaction flask was added the compound 5-2 (670 mg, 1.02 mmol, 1.0 eq), 4- (4, 5-tetramethyl-1, 3, 2-dioxaarene-2-yl) naphthalen-2-ol (411.4 mg, 1.6 mmol,1.5 equivalents), chlorine (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl ] ]Palladium (II) (79.7 mg, 0.1 mmol, 0.1 eq), potassium phosphate (431.0 mg, 2.03 mmol, 2.0 eq) and tetrahydrofuran/water (10/1, 7.7 ml), the resulting mixture was reacted at 60 ℃ for 3 hours, the course of the reaction being monitored by liquid quality. After the reaction is finished, cooling the reaction liquid to 25 ℃, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel (mobile phase eluting with a gradient of 0% → 10% methanol/dichloromethane) and the fractions were freed from the solvent by rotary evaporation under reduced pressure to give compounds 5-3 (white solid, 480 mg, 68% yield). MS (ESI, m/z) 690.3/692.3[ 2 ] M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.89(d,J＝11.6Hz,1H),7.72(d,J＝8.3Hz,1H),7.43–7.36(m,1H),7.30–7.27(m,2H),7.21–7.16(m,1H),7.14–7.09(m,1H),5.84–5.66(m,1H),5.14–5.06(m,1H),5.02–4.86(m,2H),4.53–4.39(m,3H),4.36–4.30(m,1H),4.10–3.98(m,1H),3.81(d,J＝11.9Hz,1H),3.68–3.53(m,1H),2.61(t,J＝7.4Hz,2H),2.40(t,J＝7.8Hz,2H),2.27(s,3H),2.14–1.79(m,9H),1.61–1.54(m,2H),1.51–1.46(m,9H)。

And 4, step 4:

chiral resolution of the compound 5-3 (480 mg) obtained in step 3 by preparative chiral high performance liquid chromatography: CHIRAL column CHIRAL ART amide-SA, 3 × 25 cm, 5 microns; mobile phase A: n-hexane (10 mmol/l ammonia), mobile phase B: isopropyl alcohol; flow rate: 40 ml/min; elution was performed with 30% phase B in 30 minutes, detector UV 220/254 nm. Two products were obtained, a shorter retention time (10.17 minutes) product being compound 5-3a (white solid, 180 mg, 38% recovery), compound 5-3a: MS (ESI, m/z) 690.3/692.3[ m ] +H ] ⁺ (ii) a The product at longer retention time (17.32 min) was compound 5-3b (white solid, 160 mg, 33.3% recovery), compound 5-3b: MS (ESI, m/z) 690.3/692.3[ m ] +H] ⁺ 。

And 5:

compound 5-3a (180 mg, 0.25 mmol, 1.0 equiv.) and N-methylmorpholine oxide (43.5 mg, 0.37 mmol, 1.5 equiv.) are dissolved in acetone/water (4/1, 5 mL) with stirring at 25 deg.C, followed by the slow addition of potassium osmate (18.26 mg, 0.05 mmol, 0.2 equiv.) at 0 deg.C. The reaction solution reacts for 3 hours under the condition of stirring at 0 ℃, and the reaction process is monitored by liquid quality. After the reaction, the reaction solution was purified by a reverse phase flash chromatography column (C18 column), and eluted with a mobile phase of 5% → 95% methanol/water (0.1% ammonia) over 25 minutes; detector UV254 nm; compound 5-4a was obtained (white solid, 155 mg, 82% yield). MS (ESI, m/z) 724.3/726.3[ 2 ], [ M + H ]] ⁺

Step 6:

compound 5-4a (75.0 mg, 0.098 mmol, 1.0 eq) was dissolved in acetonitrile/water (4/1,5 ml) with stirring at 25 degrees celsius, followed by the addition of sodium periodate (132.9 mg, 0.62 mmol, 6.0 eq). The reaction solution reacts for 1 hour under the stirring condition of 25 ℃, and the reaction process is monitored by liquid quality. After the reaction was completed, the reaction solution was poured into 20 ml of water, followed by extraction with dichloromethane (10 ml × 3 times), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product 5-5a (80 mg, yield 93%) which was directly used in the next reaction. MS (ESI, m/z) 692.3/694.3[ M ] +H ] ⁺ 。

And 7:

under the condition of stirring at 25 ℃ under the protection of nitrogen, the compound 1-4 (4)9.8 mg, 0.09 mmol, 1.0 eq) and sodium cyanoborohydride (18.4 mg, 0.28 mmol, 3.0 eq) were dissolved in 4 ml of methanol, followed by addition of compound 5-5a (80.0 mg, 0.09 mmol, 1.0 eq). The mixture was allowed to react for 2 hours at 25 ℃ with stirring, and the reaction was monitored by liquid mass. After the reaction was completed, the reaction solution was purified by preparative silica gel thin layer chromatography (developing solvent system: dichloromethane/methanol = 8/1) to obtain compound 5-6a (yellow oil, 80 mg, yield 75%). MS (ESI, m/z) 1089.5/1091.5[ 2 ], [ M + H ]] ⁺ 。

And 8:

compound 5-6a (80.0 mg, 0.073 mmol, 1.00 eq) was dissolved in 3 ml of dichloromethane with stirring at 25 ℃, the reaction was brought to 0 ℃ and 1 ml of trifluoroacetic acid was added dropwise. The reaction solution reacts for 1 hour under the stirring condition of 25 ℃, and the reaction process is monitored by liquid quality. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. The obtained crude product is purified by high performance liquid phase, and the preparation conditions are as follows: high-pressure reversed-phase Column XSelect CSH Prep C18 OBD Column,19 × 250 mm, 5 μm; a mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution with 5% → 20% mobile phase B for 10 minutes; detector UV254/220 nm; compound 5a was obtained (yellow solid, 20 mg, 25% yield). MS (ESI, m/z) 989.5/991.5[ m + H ] ] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.53(s,1H),8.15(d,J＝1.7Hz,1H),7.77(d,J＝8.3Hz,1H),7.57–7.52(m,1H),7.46–7.40(m,1H),7.29(d,J＝2.4Hz,1H),7.25–7.18(m,2H),7.06–7.03(m,3H),5.09–5.01(m,2H),4.57–4.53(m,2H),4.45(d,J＝11.9Hz,1H),4.29(d,J＝11.5Hz,1H),3.72–3.66(m,2H),3.46–3.33(m,4H),3.19(t,J＝7.6Hz,2H),3.00(t,J＝7.6Hz,2H),2.88–2.39(m,18H),2.21–19.7(m,6H),1.75–1.61(m,8H)； ¹⁹ F NMR(282MHz,CD ₃ OD)δ-123.20。

Step 5':

compounds 5-3b (160 mg, 0.22 mmol, 1.0 equiv.) and N-methylmorpholine oxide (40.73 mg, 0.35 mmol, 1.5 equiv.) were dissolved in acetone/water (4/1, 5 mL) with stirring at 25 deg.C, followed by slow addition of potassium osmate (17.1 mg, 0.05 mmol, 0.2 equiv.) at 0 deg.C. The reaction solution reacts for 3 hours under the condition of stirring at 0 ℃, and the reaction process is monitored by liquid quality. After the reaction, the reaction solution was purified by a reverse phase flash column (C18 column), and eluted with a 5% → 95% methanol/water (0.1% ammonia) mobile phase within 20 minutes; detector UV254 nm; compound 5-4b was obtained (white solid, 145 mg, 86% yield). MS (ESI, m/z): 724.3/726.3[ M ] +H] ⁺

Step 6':

compound 5-4b (75.0 mg, 0.098 mmol, 1.0 eq) was dissolved in acetonitrile/water (4/1, 5 ml) with stirring at 25 ℃, followed by the addition of sodium periodate (132.9 mg, 0.62 mmol, 6.0 eq). The reaction solution reacts for 1 hour under the stirring condition of 25 ℃, and the reaction process is monitored by liquid quality. After the reaction was completed, the reaction solution was poured into 20 ml of water, extracted with dichloromethane (10 ml × 3), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude product 5-5b (80 mg, yield 93%) which was directly used in the next reaction. MS (ESI, m/z) 692.3/694.3[ M ] +H ] ⁺ 。

Step 7':

under nitrogen protection stirring at 25 ℃, compounds 1-4 (49.8 mg, 0.09 mmol, 1.0 equivalent) and cyanoborohydride are addedSodium chloride (18.4 mg, 0.28 mmol, 3.0 equiv.) was dissolved in 4 ml of methanol, followed by the addition of compound 5-5b (80.0 mg, 0.09 mmol, 1.0 equiv.). The mixture was allowed to react for 2 hours at 25 ℃ with stirring, and the progress of the reaction was monitored by liquid mass. After the reaction was completed, the reaction solution was purified by preparative silica gel thin layer chromatography (developing solvent system: dichloromethane/methanol = 8/1) to obtain compound 5-6b (yellow oil, 80 mg, yield 75%). MS (ESI, m/z) 1089.5/1091.5[ 2 ], [ M + H ]] ⁺ 。

Step 8':

compound 5-6b (75.0 mg, 1.00 eq) was dissolved in 3 ml of dichloromethane with stirring at 25 degrees celsius, the reaction was brought to 0 degrees celsius, and 1 ml of trifluoroacetic acid was added dropwise. The reaction solution reacts for 1 hour under the condition of stirring at 25 ℃, and the reaction process is monitored by liquid quality. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. The obtained crude product is purified by high performance liquid phase, and the preparation conditions are as follows: high-pressure reversed-phase Column XSelect CSH Prep C18 OBD Column,19x250 mm, 5 micron; a mobile phase A: water (0.05% trifluoroacetic acid), mobile phase B methanol; flow rate: 25 ml/min; elution with 27% → 54% mobile phase B for 7 minutes; detector UV254/220 nm; compound 5b (yellow solid, 12.6 mg) was obtained. MS (ESI, m/z) 989.5/991.5[ m + H ] ] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.51(s,2H),8.15(d,J＝1.6Hz,1H),7.77(d,J＝8.3Hz,1H),7.59–7.51(m,1H),7.47–7.39(m,1H),7.29(d,J＝2.4Hz,1H),7.26–7.15(m,2H),7.09–7.01(m,3H),5.11–5.00(m,2H),4.60–4.51(m,2H),4.51–4.39(m,1H),4.36–4.27(m,1H),3.75–3.65(m,2H),3.49–3.40(m,1H),3.38–3.35(m,1H),3.25–3.17(m,2H),3.07–2.98(m,2H),2.88–2.36(m,20H),2.27–2.03(m,6H),1.78–1.60(m,8H)； ¹⁹ F NMR(282MHz,CD ₃ OD)δ-123.64。

Example 6 (Synthesis method I)

4- ((6- (4- (6- ((3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) hexyl) piperazin-1-yl) hexyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione tetra-trifluoroacetate 6

Step 1:

to a 250 ml single neck flask were added N- (6-bromohexyl) phthalimide acid (5.0 g, 16.1 mmol, 1.0 eq), 1-tert-butoxycarbonylpiperazine (3.3 g, 17.7 mmol, 1.1 eq), acetonitrile (100 ml) and N, N-diisopropylethylamine (4.17 g, 32.2 mmol, 2.0 eq) in that order with stirring at 25 degrees celsius under nitrogen. The resulting mixture was reacted at 80 ℃ for 16 hours. The course of the reaction is monitored by thin-layer chromatography (R) _f =0.7, dichloromethane/methanol = 10/1). After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 6-1 (brown oil, 4.4 g, 69% yield). MS (ESI, m/z) 416.3[ m + H ], [ M ], [ phi ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.88–7.83(m,2H),7.76–7.71(m,2H),3.72–3.67(m,2H),3.46–3.43(m,4H),2.40–2.32(m,6H),1.72–1.36(m,17H)。

Step 2:

compound 6-1 (4.4 g, 10.1 mmol, 1.0 eq) was dissolved in 100 ml ethanol with stirring at 25 ℃ and hydrazine hydrate (3.46 g, 55.3 mmol, 5.5 eq) was added. The obtained mixed system reacts for 16 hours at 80 ℃, and the reaction process is monitored by liquid quality. After the reaction is finished, the reaction is carried out The solution is cooled to 0 ℃, insoluble substances are removed by filtration, the filter cake is washed three times by 150 ml of ethanol, the filtrates are combined and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 20% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 6-2 (colorless oil, 2.3 g, 76% yield). MS (ESI, m/z) 286.2[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ3.44–3.42(m,4H),2.70–2.67(m,2H),2.38–2.31(m,6H),1.53–1.3 0(m,17H)。

And step 3:

to a 100 ml single neck flask were added 6-2 (1.1 g, 3.6 mmol, 1.05 eq), 2- (2, 6 oxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (1 g, 3.4 mmol, 1.0 eq), N-methylpyrrolidone (10 ml) and N, N-diisopropylethylamine (889 mg, 6.9 mmol, 2.0 eq) sequentially with stirring at 25 degrees celsius under nitrogen. The obtained mixture is heated to 90 ℃ to react for 2 hours, and the reaction process is monitored by liquid quality. After the reaction was completed, the reaction solution was cooled to 25 ℃. The reaction solution was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 50% methanol/water (0.1% trifluoroacetic acid) mobile phase over 25 minutes; detector UV254 nm; compound 6-3 was obtained (yellow solid, 360 mg, 18% yield). MS (ESI, m/z) 542.3[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.25(s,1H),7.51–7.47(m,1H),7.09(d,J＝7.1Hz,1H),6.88(d,J＝8.5Hz,1H),6.24–6.21(m,1H),4.93–4.89(m,1H),3.46–3.43(m,4H),3.29–3.24(m,2H),2.91–2.69(m,3H),2.39–2.33(m,6H),2.16–2.01(m,1H),1.71–1.64(m,3H),1.54–1.25(m,14H)。

And 4, step 4:

stirring at 0 deg.CTo a 25 ml single-neck flask were added 6-3 (360 mg, 0.6 mmol, 1.0 eq.), dichloromethane (3 ml) and trifluoroacetic acid (1 ml) in that order. The resulting mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid quality. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product, and the obtained crude product 6-4 (350 mg, 95%) was used directly in the next step. MS (ESI, m/z) 442.3[ 2 ], [ M ] +H] ⁺ 。

And 5:

6-hepten-1-ol (1.0 g, 8.3 mmol, 1.0 equiv.) is dissolved in 30 ml of dichloromethane with stirring at 0 deg.C, and then 1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy-3 (1H) -one (7.43 g, 17.5 mmol, 2.0 equiv.) is added in portions. The resulting mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by thin layer chromatography. After the reaction was over, the reaction was quenched with saturated sodium bisulfite/saturated sodium thiosulfate (30 ml/30 ml), the mixture was extracted with dichloromethane (50 ml x 3), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 20% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 6-5 (colorless oil, 680 mg, 46% yield). ¹ H NMR(400MHz,DMSO-d ₆ )δ9.67–9.66(m,1H),5.84–5.74(m,1H),5.04–4.93(m,2H),2.45–2.41(m,2H),2.05–1.99(m,2H),1.61–1.49(m,2H),1.39–1.31(m,2H)。

Step 6:

compounds 2-9a (150.0 mg.0.24 mmol, 1.0 eq.), acetic acid (21.53 mg, 0.36 mmol, 1.5 eq.), sodium cyanoborohydride (45.1 mg, 0.72 mmol) were stirred at 25 deg.C3.0 equivalents) was dissolved in 4 ml of methanol, and then compound 6-5 (63.9 mg, 0.36 mmol, 1.5 equivalents) was slowly added dropwise. The mixture was allowed to react for 2 hours at 25 ℃ with stirring, and the reaction was monitored by liquid mass. After the reaction was completed, the reaction solution was purified by preparative silica gel thin layer chromatography (developing solvent system: dichloromethane/methanol = 8/1) to obtain compound 6-6a (white solid, 150 mg, yield 86%). MS (ESI, m/z) 692.3/694.3[ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.73–7.69(m,2H),7.39–7.35(m,1H),7.30(d,J＝2.4Hz,1H),7.24(d,J＝8.3Hz,1H),7.18–7.14(m,2H),5.74–5.63(m,1H),4.95–4.87(m,2H),4.48–4.43(m,2H),3.86–3.84(m,4H),3.68–3.65(m,4H),3.01–2.97m,2H),2.69–2.65(m,2H),2.51(s,3H),2.19–2.15(m,2H),1.96–1.91(m,2H),1.60–1.54(m,2H),1.50(s,9H),1.32–1.25(m,2H),1.21–1.13(m,2H)。

And 7:

compounds 6-6a (100 mg, 0.137 mmol, 1.0 equiv.) and N-methylmorpholine oxide (25.4 mg, 0.21 mmol, 1.5 equiv.) were dissolved in acetone/water (4/1, 5 mL) with stirring at 25 deg.C under nitrogen, followed by slow addition of potassium osmate (5.3 mg, 0.014 mmol, 0.1 equiv.) at 0 deg.C. The obtained mixture reacts for 5 hours under the condition of stirring at the temperature of 0 ℃, and the reaction process is monitored by liquid quality. After the reaction, the reaction solution was purified by a reverse phase flash column (C18 column), and eluted with a 5% → 95% methanol/water (0.1% ammonia) mobile phase in 25 minutes; detector UV254 nm; compound 6-7a was obtained (yellow oil, 77 mg, 73% yield). MS (ESI, m/z) 726.3/728.3[ 2 ] M + H ] ⁺ 。

And 8:

stirring at 25 deg.CNext, compound 6-7a (75.0 mg, 0.098 mmol, 1.0 eq) was dissolved in acetonitrile/water (4/1, 5 ml) followed by the addition of sodium periodate (132.5 mg, 0.62 mmol, 6.0 eq). The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid mass. After the reaction was completed, the reaction solution was poured into 10 ml of water, extracted with dichloromethane (10 ml × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude product 6-8a (70 mg, yield 92%) which was directly used in the next reaction. MS (ESI, m/z) 694.3/696.3[ M ] +H] ⁺ 。

And step 9:

compound 6-4 (51.5 mg, 0.09 mmol, 1.0 eq), sodium cyanoborohydride (18.0 mg, 0.27 mmol, 3.0 eq) was dissolved in 3 ml of methanol at 25 ℃ with stirring under nitrogen protection, followed by the addition of compound 6-8a (70.0 mg, 0.09 mmol, 1.0 eq). The resulting mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid quality. After the reaction was completed, the reaction solution was directly purified by preparative silica gel thin layer chromatography (developing solvent system: dichloromethane/methanol = 6/1) to obtain compound 6-9a (yellow oil, 60 mg, yield 52%). MS (ESI, m/z) 1119.5/1121.5[ 2 ] M + H ] ⁺ 。

Step 10:

to a 25 ml single-neck flask were added 6-9a (50.0 mg, 0.042 mmol, 1.00 eq), dichloromethane (3 ml) and trifluoroacetic acid (1 ml) sequentially with stirring at 0 ℃. After the addition, the reaction was carried out for 1 hour under stirring at 25 ℃ and the reaction process was monitored by liquid quality. After the reaction is finished, concentrating the reaction liquid under reduced pressure to obtain a crude product. The obtained crude product is purified by high performance liquid phase, and the preparation conditions are as follows: high-pressure reversed-phase Column XSelect CSH Prep C18 OBD Column19x250 mm, 5 microns; a mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; elution with 5% → 40% mobile phase B for 7 minutes; detector UV254/220 nm; compound 6 was obtained (yellow solid, 20 mg, 31% yield). MS (ESI, m/z) 1019.5/1021.5[ M + H ]] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.07(d,J＝1.7Hz,1H),7.79(d,J＝8.3Hz,1H),7.62–7.53(m,1H),7.49–7.41(m,1H),7.30(d,J＝2.4Hz,1H),7.26–7.21(m,2H),7.11–7.03(m,3H),5.14–5.02(m,1H),4.70–4.56(m,2H),4.24–4.08(m,4H),3.56–3.35(m,11H),3.32–2.64(m,17H),2.41–2.25(m,2H),2.20–2.05(m,1H),1.84–1.61(m,8H),1.60–1.38(m,8H)。

Example 7

4- ((7- (4- (7- ((3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) heptyl) piperazin-1-yl) heptyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione tetra-trifluoroacetate 7

Step 1

To a 100 ml round bottom flask was added sequentially potassium phthalimide (1.79 g, 9.181 mmol, 1.0 eq), acetone (30.0 ml, 387.661 mmol, 42.23 eq) and 1, 7-dibromoheptane (4.99 g, 18.36 mmol, 2.0 eq) with stirring at 25 ℃. The resulting mixture was reacted at 60 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, cooling the reaction liquid to room temperature, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 40% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 7-1 (colorless oil, 2.3 g, 77% yield). MS (ESI, m/z) 324.0/326.0 [M+H] ⁺ ； ¹ H NMR(400MHz，CDCl ₃ )δ7.87–7.82(m，2H)，7.74–7.69(m，2H)，3.70–3.66(m，2H)，3.41–3.38(m，2H)，1.88–1.81(m，2H)，1.72–1.65(m，2H)，1.47–1.34(m，6H)。

Step 2

To a 100 ml round bottom flask were added 7-1 (2.3 g, 6.739 mmol, 1.0 eq), 1-tert-butoxycarbonylpiperazine (1.45 g, 7.413 mmol, 1.1 eq), acetonitrile (30.0 ml, 542.204 mmol, 80.45 eq) and N, N-diisopropylethylamine (1.83 g, 13.478 mmol, 2.00 eq) sequentially with stirring at 25 ℃. After the addition, the reaction was stirred for 4 hours while the temperature was raised to 80 ℃. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, cooling the reaction liquid to room temperature, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 7-2 (light yellow oil, 2.5 g, 86% yield). MS (ESI, m/z) 430.3[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz，CDCl ₃ )δ7.86–7.82(m，2H)，7.73–7.69(m，2H)，3.69–3.66(m，2H)，3.44–3.41(m，4H)，2.37–2.29(m，6H)，1.71–1.64(m，2H)，1.51–1.43(m，11H)，1.36–1.27(m，6H)。

Step 3

Compound 7-2 (2.5 g, 5.529 mmol, 1.0 eq), ethanol (100.0 ml) and hydrazine hydrate (80%) (1.17 g, 22.116 mmol, 4.00 eq) were added sequentially to a 500 ml round bottom flask with stirring at 25 ℃. The resulting mixture was reacted at 80 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is cooled to room temperature, Concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 7-3 (colorless oil, 1.2 g, 72% yield). MS (ESI, m/z) 300.6[ 2 ] M + H] ⁺ ； ¹ H NMR(400MHz，CD ₃ OD)δ3.44–3.41(m，4H)，2.64–2.61(m，2H)，2.42–2.34(m，6H)，1.56–1.45(m，13H)，1.37–1.32(m，6H)。

Step 4

To a 40 ml reaction flask were added 2- (2, 6-dioxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (500.0 mg, 1.720 mmol, 1.0 eq), N-methylpyrrolidone (5.00 ml), N-diisopropylethylamine (467.89 mg, 3.44 mmol, 2.0 eq) and compound 7-3 (569.16 mg, 1.806 mmol, 1.05 eq) with stirring at 25 ℃. The temperature was raised to 90 ℃ at the end of the addition and the reaction was carried out at this temperature for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is cooled to room temperature, and a crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 7-4 (yellow solid, 200 mg, 20% yield). MS (ESI, m/z) 556.1[ 2 ] M + H] ⁺ 。

And 5:

to a 50 ml round bottom flask were added sequentially compound 7-4 (200.0 mg, 0.342 mmol, 1.0 eq), dichloromethane (6.00 ml) and trifluoroacetic acid (2.00 ml) with stirring at 0 ℃. The resulting mixture was reacted at 25 ℃ for 1 hour. The course of the reaction is monitored by the liquid quality. After the reaction is finished, decompressing and concentrating to obtain a compound 7-5 crude product (yellow solid, 120 mg) and the crude product was used directly in the next step. MS (ESI, m/z) 456.8[ 2 ], [ M ] +H] ⁺ 。

Step 6

7-Octen-1-ol (2.0 g, 14.819 mmol, 1.0 equiv.) was dissolved in dichloromethane (200.00 ml) with stirring at 25 deg.C, to which was then added 1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy-3 (1H) -one (13.23 g, 29.638 mmol, 2.00 equiv.) in portions. The resulting mixture was reacted for 2 hours at 25 ℃ with stirring. The course of the reaction was monitored by thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 30% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 7-6 (bright yellow oil, 1.5 g, 80% yield). ¹ H NMR(400MHz，DMSO-d ₆ )δ9.66–9.65(m，1H)，5.84–5.74(m，1H)，5.03–4.92(m，2H)，2.44–2.40(m，2H)，2.04–1.98(m，2H)，1.56–1.49(m，2H)，1.39–1.22(m，4H)。

Step 7

To an 8 ml reaction flask were added, under stirring at 25 ℃, compounds 2-9a (120.0 mg, 0.191 mmol, 1.0 eq), methanol (2.00 ml), acetic acid (17.23 mg, 0.287 mmol, 1.50 eq) and sodium cyanoborohydride (36.05 mg, 0.573 mmol, 3.00 eq) in that order, and finally compounds 7-6 (38.11 mg, 0.287 mmol, 1.50 eq). The resulting mixture was reacted at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the fractions obtained were passed through The solvent was removed by rotary evaporation under reduced pressure to give compound 7-7a (brown solid, 110 mg, 81% yield). MS (ESI, m/z) 706.2/708.2[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(400MHz，CDCl ₃ )δ7.74–7.72(m，2H)，7.43–7.39(m，1H)，7.29–7.27(m，2H)，7.23–7.19(m，1H)，7.07(d，J＝2.4Hz，1H)，5.82–5.71(m，1H)，4.99–4.88(m，2H)，4.50–4.47(m，2H)，3.82–3.80(m，4H)，3.68–3.66(m，4H)，2.61–2.57(m，2H)，2.40–2.36(m，2H)，2.27(s，3H)，2.08–1.96(m，4H)，1.51(s，9H)，1.48–1.25(m，8H)。

Step 8

To a 50 ml round bottom flask were added sequentially compound 7-7a (99.00 mg, 0.133 mmol, 1.00 eq), acetone (3.20 ml)/water (0.80 ml), potassium osmate dihydrate (5.16 mg, 0.013 mmol, 0.10 eq) and N-methylmorpholine oxide (24.63 mg, 0.2 mmol, 1.50 eq) with stirring at 0 degrees celsius. The resulting mixture was reacted at 0 ℃ for 2 hours. The reaction process is monitored by liquid quality. After the reaction was completed, the reaction solution was heated to 25 ℃, concentrated, purified by a reverse phase flash column (C18 column), and eluted with 10% → 95% acetonitrile/water mobile phase (0.1% ammonia) within 20 minutes; detector UV254 nm; compound 7-8a was obtained (yellow oil, 80 mg, 81% yield). MS (ESI, m/z) 740.5/742.5[ M + H ]] ⁺ 。

Step 9

Compound 7-8a (80.0 mg, 0.103 mmol, 1.0 eq), acetonitrile (4.00 ml)/water (1.00 ml) was added to a 50 ml round bottom flask with stirring at 25 degrees celsius, then sodium periodate (138.68 mg, 0.618 mmol, 6.0 eq) was added thereto in portions. The resulting mixture was reacted at 25 deg.C It should be 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was poured into 10 ml of water and extracted with dichloromethane (10 ml × 3). The combined organic phases were washed with 20 ml of saturated brine, the washed organic phases were dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure to give 7-9a as a crude product (yellow solid, 60 mg, 82% yield) which was used directly in the next step. MS (ESI, m/z) 708.3/710.3[ m ] +H] ⁺ 。

Step 10

To a 50 ml round bottom flask, compound 7-5 (51.46 mg, 0.107 mmol, 1.00 eq), sodium cyanoborohydride (21.29 mg, 0.321 mmol, 3.00 eq) and methanol (3 ml) were added with stirring at 25 ℃, and finally compound 7-9a (80.0 mg, 0.107 mmol, 1.0 eq) was added slowly to the reaction system above. The resulting mixture was reacted at 25 ℃ for 2 hours. The reaction process is monitored by liquid chromatography and thin layer chromatography, and after the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 7-10a (yellow solid, 80.0 mg, 39% yield). MS (ESI, m/z) 1147.6/1149.6[ 2 ] M + H ] ⁺ 。

Step 11

To a 25 ml single-neck flask were added sequentially compound 7-10a (80.0 mg, 0.042 mmol, 1.00 eq), dichloromethane (3 ml) and trifluoroacetic acid (1 ml) with stirring at 0 ℃. The reaction solution reacts for 1 hour under the stirring condition of 25 ℃, and the reaction process is monitored by liquid quality. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. The obtained crude product is purified by high performance liquid phase, and the preparation conditions are as follows: height ofPressure reversed phase Column XSelect CSH Prep C18 OBD Column,19X250 mm, 5 micron; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B acetonitrile; flow rate: 25 ml/min; elution with 5% → 40% mobile phase B for 10 minutes; detector UV254/220 nm; compound 7 was obtained (yellow solid, 61.2 mg, 37% yield). MS (ESI, m/z) 1047.5/1049.5[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(400MHz，CD ₃ OD)δ8.04(d，J＝1.6Hz，1H)，7.77–7.75(m，1H)，7.56–7.52(m，1H)，7.44–7.40(m，1H)，7.28(d，J＝2.4Hz，1H)，7.22–7.17(m，2H)，7.05–7.03(m，3H)，5.07–5.02(m，1H)，4.62–4.58(m，2H)，4.14–4.12(m，4H)，3.52–3.34(m，12H)，3.29–2.64(m，16H)，2.30–2.28(m，2H)，2.12–2.06(m，1H)，1.74–1.66(m，8H)，1.43–1.29(m，12H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-77.18,-123.05。

Example 8

4- (4- (4- (4- ((3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butoxy) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione diformate salt 8

Step 1

To a solution of compound 1-tert-butoxycarbonylpiperazine (5.0 g, 25.5 mmol, 1.0 eq) and 4-chloro-1-butanol (3.5 g, 0.03 mmol, 1.2 eq) in acetonitrile (50.0 ml) was added sodium bicarbonate (9.0 g, 102.0 mmol, 4.0 eq) and sodium iodide (2.0 g, 12.8 mmol, 0.5 eq) with stirring at 25 ℃. The resulting mixture was reacted at 90 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the reaction liquid is cooled to room temperature, and a crude product is obtained by decompression and concentration. Adding 100.0 parts of the crude product Ml of water, extraction with ethyl acetate (100 ml x 3), combination of the organic phases, drying of the organic phases with anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure to give the crude product, purification of the crude product by silica gel column chromatography, elution with a mobile phase gradient of 0% → 10% methanol/dichloromethane, removal of the solvent from the fractions obtained by rotary evaporation under reduced pressure to give compound 8-1 (yellow liquid, 2.6 g, 36% yield). MS (ESI, m/z) 259.1[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ3.60–3.54(m,2H),3.50–3.42(m,4H),2.50–2.38(m,6H),1.74–1.65(m,4H),1.46(s,9H)。

Step 2

To a solution of compound 8-1 (819.0 mg, 3.2 mmol, 2.2 eq) in tetrahydrofuran (1.0 ml) was added dropwise lithium bis-trimethylsilylamido (1 mol/l tetrahydrofuran solution) (3.2 ml, 3.2 mmol, 2.2 eq) at 25 ℃ with stirring under nitrogen. After the addition of the resulting mixture, the reaction was continued for 1 hour at 60 ℃ and then a solution of 2- (2, 6-dioxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (398.0 mg, 1.4 mmol, 1.0 eq) in toluene (6.0 ml) was added dropwise to the reaction solution. The resulting mixture was stirred at 100 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the reaction liquid is cooled to room temperature, and a crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and give compound 8-2 (white solid, 80 mg, 11% yield). MS (ESI, m/z) 515.3[ 2 ], [ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ8.24(s,1H),7.72–7.67(m,1H),7.46(d,J＝7.2Hz,1H),7.21(d,J＝8.2Hz,1H),4.99–4.93(m,1H),4.28–4.17(m,2H),3.52–3.40(m,4H),2.95–2.72(m,3H),2.52–2.36(m,4H),2.17–2.12(m,1H),1.99–1.89(m,2H),1.81–1.65(m,4H),1.45(s,9H)。

Step 3

To a solution of compound 8-2 (110.0 mg, 0.20 mmol, 1.0 eq) in dichloromethane (4.0 ml) was added trifluoroacetic acid (1.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave crude product 8-3 (off-white solid, 150.0 mg). MS (ESI, m/z) 415.4[ 2 ], [ M ] +H] ⁺ 。

Step 4

To the reaction flask were added compound 2-9a (200.0 mg, 0.32 mmol, 1.0 equivalent), acetic acid (28.7 mg, 0.48 mmol, 1.5 equivalents), sodium cyanoborohydride (60.0 mg, 0.96 mmol, 3.0 equivalents) and methanol (3.0 ml) in this order with stirring at 25 ℃, and then a solution of 4-pentenal (40.2 mg, 0.48 mmol, 1.5 equivalents) in methanol (0.5 ml) was added dropwise. The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 8-4a (white solid, 150.0 mg, yield 71%) was obtained. MS (ESI, m/z) 664.4/666.4[ 2 ] M + H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.76–7.70(m,2H),7.43–7.38(m,1H),7.32(d,J＝2.4Hz,1H),7.25–7.24(m,1H),7.22–7.16(m,1H),7.14(d,J＝2.4Hz,1H),5.79–5.57(m,1H),5.01–4.92(m,2H),4.53–4.43(m,2H),3.90–3.76(m,4H),3.69–3.64(m,4H),2.96(s,2H),2.69(s,2H),2.52(s,3H),2.21(s,2H),2.06–1.98(m,2H),1.78–1.69(m,2H),1.51(s,9H)。

Step 5

To a solution of compounds 8-4a (65.0 mg, 0.09 mmol, 1.0 eq) in acetone/water (4/1, 2.0 mL) was added N-methylmorpholine oxide (16.3 mg, 0.14 mmol, 1.5 eq) and potassium osmate dihydrate (6.9 mg, 0.02 mmol, 0.2 eq) with stirring at 0 deg.C. The resulting mixture was stirred at 0 ℃ for 5 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was directly purified by a reverse phase flash column (C18 column), and eluted with 30% → 95% methanol/water mobile phase (0.1% ammonia) within 20 minutes; detector UV254 nm; compound 8-5a was obtained (pale yellow solid, 66.0 mg, 96% yield). MS (ESI, m/z) 698.4/700.3[ M + H ]] ⁺ 。

Step 6

To a solution of compound 8-5a (50.0 mg, 0.07 mmol, 1.0 equiv.) in acetonitrile/water (4/1, 1.0 ml) was added sodium periodate (91.9 mg, 0.4 mmol, 6.0 equiv) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, water (10.0 ml), dichloromethane (10.0 ml x 3) were added to the reaction solution for extraction, the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude 8-6a (pale yellow oily liquid, 55.0 mg) which was used in the next reaction without further purification. MS (ESI, m/z) 666.2/668.2[ 2 ] M + H ] ⁺ 。

Step 7

To compound 8-3 (36.6 mg, 0.07 mmol,1.0 eq) in methanol (2.0 ml) was added sodium cyanoborohydride (13.1 mg, 0.20 mmol, 3.0 eq) and then a solution of compound 8-6a (55.0 mg, 0.07 mmol, 1.0 eq) in methanol (0.5 ml) was added dropwise. The resulting mixture was stirred at 25 ℃ for an additional 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the obtained fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 8-7a (colorless oily liquid, 50.0 mg, yield 67%) was obtained. MS (ESI, m/z) 1064.4/1066.4[ m + H ]] ⁺ 。

Step 8

To a solution of compound 8-7a (65.0 mg, 0.06 mmol, 1.0 eq) in dichloromethane (4.0 ml) was added trifluoroacetic acid (1.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 19% b phase gradient over 10 minutes; detector UV 254/220 nm; compound 8 was obtained (white solid, 15.0 mg, 24% yield). MS (ESI, m/z) 964.4/966.4[ m ] +H ] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ11.09(s,1H),8.25–8.14(m,2H),,7.97(s,1H),7.80(s,2H),7.54–7.40(m,3H),7.29(d,J＝2.4Hz,1H),7.25–7.18(m,2H),7.07(d,J＝2.4Hz,1H),5.11–5.05(m,1H),4.43–4.36(s,2H),4.20(t,J＝6.2Hz,2H),3.89–3.81(m,5H),3.10–2.97(m,4H),2.93–2.87(m,1H),2.75–2.72(m,1H),2.63–2.60(m,1H),2.43–2.14(m,18H),2.09–1.86(m,4H),1.80–1.70(m,2H),1.63–1.54(m,2H),1.46–1.33(m,4H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-122.27。

Example 9

4- ((3- (4- (3- ((3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) propyl) piperazin-1-yl) propyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione diformate 9a;4- ((3- (4- (3- ((3- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) propyl) piperazin-1-yl) propyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione formate 9b

Step 1

To a mixture of compound 3- (methylamino) -1-propanol (5.0 g, 56.1 mmol, 1.0 eq), dichloromethane (50.0 ml), and N, N-diisopropylethylamine (21.7 g, 168.3 mmol, 3.0 eq) was added di-tert-butyl dicarbonate (15.9 g, 72.9 mmol, 1.3 eq) dropwise with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 40% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 9-1 (colorless oily liquid, 10.1 g, yield 95%). MS (ESI, m/z) 190.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ3.57(s,2H),3.40(t,J＝6.2Hz,2H),2.85(s,3H),1.78–1.64(m,2H),1.48(s,9H)。

Step 2

To a solution of compound 9-1 (3.0 g, 15.9 mmol, 1.0 eq) in dichloromethane (30.0 ml) was added 1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy-3 (1H) -one (13.44 g, 31.8 mmol, 2.0 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for an additional 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 30% ethyl acetate/petroleum ether mobile phase gradient, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 9-2 (colorless oily liquid, 1.4 g, yield 47%) was obtained. MS (ESI, m/z) 188.2[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ9.83(t,J＝1.7Hz,1H),3.57(t,J＝6.6Hz,2H),2.89(s,3H),2.70(m,2H),1.47(s,9H)。

Step 3

To a 50 ml three-necked flask was added tert-butyl 4- (3-aminopropyl) piperazine-1-carboxylate (1.0 g, 4.1 mmol, 1.0 eq), N-methylpyrrolidinone (10.0 ml), 2- (2, 6-dioxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (1.1 g, 4.1 mmol, 1.0 eq) and N, N-diisopropylethylamine (0.94 g, 7.2 mmol, 1.8 eq) at 25 degrees celsius with stirring under nitrogen. The resulting mixture was stirred at 90 ℃ for a further 3 hours, the course of the reaction being monitored by liquid chromatography and thin-layer chromatography. After completion of the reaction, the reaction mixture was purified by reverse phase chromatography (C18), eluted with a mobile phase of 5% → 95% acetonitrile/water (0.1% trifluoroacetic acid) within 35 minutes, and then subjected to a detector UV 254/220 nm. Compound 9-3 was obtained (yellow solid, 650.0 mg, 32% yield). MS (ESI, m/z) 500.4[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ8.23(s,1H),7.53–7.48(m,1H),7.12–7.10(m,1H),6.94(d,J＝8.5Hz,1H),6.60(t,J＝5.6Hz,1H),4.96–4.90(m,1H),3.52–3.49(m,4H),3.42–3.36(m,2H),2.97–2.68(m,3H),2.50(t,J＝6.6Hz,2H),2.45–2.41(m,4H),2.23–2.04(m,1H),1.89–1.83(m,2H),1.48(s,9H)。

Step 4

To a solution of compound 9-3 (600.0 mg, 1.1 mmol, 1.0 eq) in dichloromethane (6.0 ml) was added trifluoroacetic acid (1.5 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for an additional 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave crude product 9-4 (yellow solid, 700.0 mg). The compound was used in the next synthesis without further purification. MS (ESI, m/z) 400.1[ 2 ] M + H] ⁺ 。

Step 5

To a solution of compound 9-4 (600.0 mg, 1.1 mmol, 1.0 eq) in methanol (7.0 ml) was added compound 9-2 (355.3 mg, 1.8 mmol, 1.2 eq) and sodium cyanoborohydride (198.7 mg, 3.0 mmol, 2.0 eq) sequentially with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase chromatography (C18) eluting with 5% → 70% acetonitrile/water (0.1% trifluoroacetic acid) mobile phase over 20 minutes with detector UV 254/220 nm to give compound 9-5 (yellow solid, 435 mg, 90% yield). MS (ESI, m/z) 571.2[ 2 ], [ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ10.63(s,1H),7.55–7.50(m,2H),7.13(d,J＝7.0Hz,1H),6.90(d,J＝8.6Hz,1H),4.99–4.93(m,1H),3.70–3.12(m,8H),3.07–2.51(m,14H),2.20–1.92(m,3H),1.86(s,2H),1.49(s,9H)。

Step 6

To a solution of compound 9-5 (400.0 mg, 0.7 mmol, 1.0 eq) in dichloromethane (9.0 ml) was added trifluoroacetic acid (3.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave crude product 9-6 (yellow solid, 400.0 mg). The compound was used in the next synthesis without further purification. MS (ESI, m/z) 471.2[ 2 ], [ M ] +H] ⁺ 。

Step 7

To the reaction flask were added compounds 9-6 (116.6 mg, 0.2 mmol, 1.2 equivalents), sodium acetate (25.4 mg, 0.3 mmol, 1.5 equivalents), acetic acid (12.4 mg, 0.2 mmol, 1.0 equivalents), sodium cyanoborohydride (38.9 mg, 0.6 mmol, 3.0 equivalents) and methanol (2.0 ml) in that order with stirring at 25 degrees celsius, and a solution of compounds 1-11a (114.0 mg, 0.2 mmol, 1.0 equivalent) in methanol (0.5 ml) was added to completion. The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 9-7a (pale yellow solid, 100.0 mg, yield 49%) was obtained. MS (ESI, m/z) 1035.4/1037.4[ 2 ] M + H ] ⁺ 。

Step 8

To a solution of compound 9-7a (50.0 mg, 0.05 mmol, 1.0 eq) in dichloromethane (2.0 ml) was added trifluoroacetic acid (0.5 ml) with stirring at 25 deg.C. The resulting mixture was stirred at 25 ℃ for an additional 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; a mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 25% b phase gradient over 7 minutes; detector UV 254/220 nm; compound 9a was obtained (yellow solid, 25 mg, 50% yield). MS (ESI, m/z) 935.4/937.4[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ11.11(s,1H),8.19(s,2H),7.98(d,J＝1.7Hz,1H),7.81(d,J＝8.2Hz,1H),7.59–7.53(m,1H),7.46–7.41(m,1H),7.29(d,J＝2.4Hz,1H),7.21(d,J＝5.0Hz,2H),7.15–6.97(m,3H),6.75(d,J＝5.9Hz,1H),5.07–5.03(m,1H),4.39–4.34(m,2H),3.88–3.86(m,5H),3.37–3.30(m,3H),3.11–3.08(m,4H),2.95–2.81(m,2H),2.64–2.54(m,4H),2.32–2.28(m,15H),2.10–1.83(m,4H),1.75–1.54(m,4H)。

Step 7'

To a reaction flask were added compounds 9-6 (116.6 mg, 0.2 mmol, 1.2 equivalents), sodium acetate (25.4 mg, 0.3 mmol, 1.5 equivalents), acetic acid (12.4 mg, 0.2 mmol, 1.0 equivalents), sodium cyanoborohydride (38.9 mg, 0.6 mmol, 3.0 equivalents) and methanol (2.0 ml) in this order with stirring at 25 ℃, and then compounds 1-11b (114 mg, 0.2 mmol, 1.0 equivalents) were added thereto. The resulting mixture was stirred at 25 ℃ for an additional 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 9-7b (pale yellow solid, 106 mg, yield 52%) was obtained. MS (ESI, m/z) 1035.4/1037.4[ 2 ], [ M + H ] ] ⁺ 。

Step 8'

To a solution of compound 9-7b (100.0 mg) in dichloromethane (2.0 ml) was added trifluoroacetic acid (0.5 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for an additional 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 25% b phase gradient over 7 minutes; detector UV 254/220 nm; compound 9b was obtained (yellow solid, 55 mg, 58% yield). MS (ESI, m/z) 935.4/937.4[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.53(s,1H),8.00(s,1H),7.75–7.71(m,1H),7.54–7.50(m,1H),7.40–7.38(m,1H),7.30–7.16(m,3H),7.08–7.00(m,3H),5.04–4.87(m,1H),4.58–4.53(m,2H),4.04–4.01(m,4H),3.38(d,J＝6.2Hz,2H),3.32–3.24(m,4H),3.13–3.05(m,2H),2.99–2.94(m,2H),2.89–2.65(m,12H),2.65–2.43(m,6H),2.21–2.17(m,2H),2.12–2.08(m,1H),1.94–1.88(m,2H),1.82–1.79(m,2H)。

Example 10

4- ((4- (4- ((3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione diformate 10a;4- ((4- (4- ((3- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione diformate 10b

Step 1

Cesium fluoride (0.81 g, 5.072 mmol, 2.0 equiv.), compound 4-1 (0.63 g, 3.804 mmol, 1.5 equiv.) and N, N-diisopropylethylamine (0.69 g, 5.072 mmol, 2.0 equiv.) were added to a solution of compounds 1-7 (1.23 g, 2.536 mmol, 1.00 equiv.) in N-methylpyrrolidinone (50 ml) at 25 degrees celsius with stirring under nitrogen. The resulting mixture was stirred at 100 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature. The reaction solution was directly purified by reverse phase flash chromatography (C18 column) and eluted with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 30 minutes; detector UV254 nm; compound 10-1 was obtained (white solid, 760 mg, 48% yield). MS (ESI, m/z) 600.3/602.3/604.3[ 2 ] M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.68(d,J＝2.0Hz,1H),5.81–5.68(m,1H),5.07–4.96(m,2H),4.51(t,J＝6.0Hz,2H),3.86–3.71(m,4H),3.69–3.55(m,4H),3.13–3.01(m,2H),2.87–2.76(m,2H),2.64(s,3H),2.34–2.19(m,2H),2.15–2.07(m,2H),1.87–1.76(m,2H),1.48(s,9H)。

Step 2

To a solution of compound 10-1 (710.00 mg, 1.142 mmol, 1.00 eq, 96.7%) in tetrahydrofuran/water (10/1, 11 ml) was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (487.30 mg, 1.714 mmol, 1.5 eq), potassium phosphate (510.54 mg, 2.285 mmol, 2.0 eq) and chlorine (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl ] at 25 ℃ in that order with stirring under nitrogen ]Palladium (II) (94.50 mg, 0.114 mmol, 0.1 eq). The resulting mixtureThe reaction was stirred at 60 ℃ for 3 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, cooling the reaction liquid to 25 ℃, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 10-2 (yellow solid, 750 mg, 98% yield). MS (ESI, m/z) 664.3/666.3[ M + H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.76–7.70(m,2H),7.44–7.38(m,1H),7.32–7.28(m,1H),7.28–7.26(m,1H),7.23–7.18(m,1H),7.10(d,J＝2.5Hz,1H),5.80–5.66(m,1H),5.05–4.87(m,2H),4.55–4.44(m,2H),3.88–3.61(m,8H),2.82–2.68(m,2H),2.60–2.46(m,2H),2.44–2.32(m,3H),2.18–2.08(m,2H),2.06–1.99(m,2H),1.71–1.62(m,2H),1.51(s,9H)。

Step 3

Chiral resolution of compound 10-2 (710 mg) by preparative chiral high performance liquid chromatography: CHIRAL column CHIRAL ART Cellulose-SC,2 × 25 cm, 5 μm; mobile phase A: n-hexane/dichloromethane =5/1, mobile phase B: ethanol; flow rate: 20 ml/min; elution was performed with 50% of phase B in 3.3 minutes, detector UV 220/230 nm. Two products were obtained, the product with a shorter retention time (6.0 minutes) being Compound 10-2a (yellow green solid, 300 mg, 43% recovery), MS (ESI, m/z): 664.3/666.3[ M ] +H ],] ⁺ (ii) a The product with longer retention time (7.4 min) is compound 10-2b (yellow green solid, 210 mg, recovery 30%), MS (ESI, m/z): 664.3/666.3[ M + H ]) ] ⁺ 。

Step 4

To compound 10-2a (300.00 mg, 0.452 mmol, 1.00 eq) in water/tetrahydrofuran was added with stirring at 0 deg.CTo a solution of pyran (1/4, 10 ml) were added N-methylmorpholine oxide (113.4 mg, 0.9 mmol, 2 equiv.) and osmium tetroxide (6.0 mg, 0.024 mmol, 0.05 equiv.). The resulting mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 30 minutes; detector UV254 nm; to obtain compound 10-3a (yellow green solid, 240 mg, yield 73%) MS (ESI, m/z): 698.2/670.2[ M ] +H] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ7.98(d,J＝1.6Hz,1H),7.76–7.72(m,1H),7.43–7.37(m,1H),7.25(d,J＝2.5Hz,1H),7.23–7.17(m,2H),7.03(d,J＝2.4Hz,1H),4.54–4.46(m,2H),4.00–3.92(m,4H),3.74–3.65(m,4H),3.59–3.51(m,1H),3.41(d,J＝5.5Hz,2H),2.67–2.59(m,2H),2.48–2.41(m,2H),2.29(s,3H),2.07–1.98(m,2H),1.75–1.55(m,4H),1.50(s,9H)1.39–1.27,(m,2H)。

Step 5

To a solution of compound 10-3a (80.00 mg, 0.115 mmol, 1.00 eq) in water/tetrahydrofuran (1/4, 5 ml) was added sodium periodate (147.04 mg, 0.687 mmol, 6.0 eq) with stirring at 0 ℃. The resulting mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, water (50 ml) was added to the reaction solution, and extracted with dichloromethane (50 ml × 3). After combination, the organic phase is dried by anhydrous sodium sulfate, filtered, and the filtrate is concentrated to obtain a crude product 10-4a. MS (ESI, m/z) 666.3/668.3[ 2 ] M + H ] ⁺ 。

Step 6

Stirring at 25 deg.C, adding into a reaction flaskCompounds 1 to 4 (71.06 mg, 0.172 mmol, 1.50 equiv), acetic acid (13.76 mg, 0.229 mmol, 2.0 equiv), sodium cyanoborohydride (10.80 mg, 0.172 mmol, 1.5 equiv, 95%) and methanol (3 ml) were added in this order, and then a solution of the above 10-4a in methanol (1 ml) was added dropwise thereto. The resulting mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the mixture is purified by preparative high performance liquid chromatography, and the purification conditions are as follows: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a phase B gradient of 35% → 61% in 7 minutes; detector UV 254/220 nm; compound 10-5a was obtained (yellow-green solid, 50 mg, 41% yield). MS (ESI, m/z) 1063.3/1065.4[ m + H ]] ⁺ 。

Step 7

To a solution of compound 10-5a (50.00 mg, 0.047 mmol, 1.00 eq) in dichloromethane (2 ml) was added trifluoroacetic acid (1 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 35% → 60% b phase gradient over 7 minutes; detector UV 254/220 nm; yield 10a (yellow-green solid, 15 mg, 32% yield). MS (ESI, m/z) 963.5/965.5[ m ] +H ] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.35(s,2H),7.91(d,J＝1.6Hz,1H),7.65(d,J＝8.3Hz,1H),7.48–7.39(m,1H),7.36–7.27(m,1H),7.19–7.07(m,3H),6.99–6.88(m,3H),4.99–4.90(m,1H),4.48(t,J＝5.8Hz,2H),4.03–3.92(m,4H),3.35–3.26(m,4H),3.26–3.22(m,4H),3.05–2.96(m,2H),2.79–2.46(m,18H),2.20–2.11(m,2H),2.05–1.93(m,1H),1.70–1.50(m,8H)。

Step 4'

To a solution of compound 10-2b (210.00 mg, 0.316 mmol, 1.00 eq) in water/tetrahydrofuran (1/4, 10 ml) was added N-methylmorpholine oxide (77.97 mg, 0.632 mmol, 2.0 eq) and osmium tetroxide (4.23 mg, 0.016 mmol, 0.05 eq) with stirring at 0 degrees celsius. The resulting mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 30 minutes; detector UV254 nm; compound 10-3b was obtained as a yellow-green solid (150 mg, 65% yield). MS (ESI, m/z) 698.2/700.2[ M + H ]] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ7.99(d,J＝1.7Hz,1H),7.74(d,J＝8.3Hz,1H),7.44–7.37(m,1H),7.25(d,J＝2.4Hz,1H),7.23–7.17(m,2H),7.03(d,J＝2.4Hz,1H),4.50(t,J＝6.3Hz,2H),3.98–3.90(m,4H),3.75–3.64(m,4H),3.60–3.52(m,1H),3.44–3.39(m,2H),2.69–2.60(m,2H),2.52–2.40(m,2H),2.34–2.28(m,3H),2.09–1.99(m,2H),1.76–1.54(m,4H),1.50(s,9H),1.39–1.27(m,2H)。

Step 5'

To a solution of compound 10-3b (80.00 mg, 0.099 mmol, 1.00 eq) in water/tetrahydrofuran (1/4,5 ml) was added sodium periodate (133.82 mg, 0.594 mmol, 6.0 eq) with stirring at 0 ℃. The resulting mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, water (50 ml) was added to the reaction solution, followed by addition of dichloro-benzene Methane extraction (50 ml x 3). After combining, the organic phase is dried by anhydrous sodium sulfate, filtered, and the filtrate is concentrated to obtain a crude product 10-4b. MS (ESI, m/z) 666.3/668.3[ 2 ] M + H] ⁺ 。

Step 6'

To a reaction flask were added compounds 1-4 (64.67 mg, 0.149 mmol, 1.5 equivalents), acetic acid (12.52 mg, 0.198 mmol, 2.0 equivalents), sodium cyanoborohydride (9.83 mg, 0.149 mmol, 1.5 equivalents) and methanol (2 ml) in this order under stirring at 25 ℃, and then a solution of compound 10-4b in methanol (1 ml) was added dropwise. The resulting mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the mixture is purified by preparative high performance liquid chromatography, and the purification conditions are as follows: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: eluting with 35% → 61% B phase gradient in 7 min; detector UV 254/220 nm; yield 10-5b (yellow-green solid, 50 mg, 47% yield). MS (ESI, m/z) 1063.5/1065.4[ m + H ]] ⁺ 。

Step 7'

Trifluoroacetic acid (1 ml) was added to a solution of compound 10-5B ((50.00 mg, 0.047 mmol, 1.00 equiv.) in dichloromethane (2 ml) with stirring at 25 deg.C, the reaction was monitored by liquid chromatography and thin layer chromatography for 1 hour at 25 deg.C, after completion of the reaction, the mixture was concentrated under reduced pressure to give a crude product which was purified by preparative high performance liquid chromatography under conditions of XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm, mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile, flow rate: 25 ml/min (ii) a Gradient: eluting with 5% → 25% B phase gradient in 7 min; detector UV 254/220 nm; yield 10b (yellow-green solid, 17.6 mg, 38% yield). MS (ESI, m/z) 963.4/965.5[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.35(s,2H),7.92(d,J＝1.6Hz,1H),7.66(d,J＝8.3Hz,1H),7.48–7.40(m,1H),7.35–7.27(m,1H),7.17(d,J＝2.4Hz,1H),7.15–7.06(m,2H),6.97–6.90(m,3H),5.01–4.88(m,1H),4.49(t,J＝5.9Hz,2H),4.08–3.92(m,4H),3.36–3.28(m,4H),3.27–3.22(m,4H),3.07–2.98(m,2H),2.88–2.39(m,18H),2.22–2.10(m,2H),2.05–1.94(m,1H),1.74–1.45(m,8H)。

Example 11

5- ((4- (4- (4- ((3- (((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione trifluoroacetate 11

Step 1

To a solution of compound 1-2 (3.5 g, 12.9 mmol, 1.1 eq) and 2- (2, 6-dioxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (3.4 g, 11.7 mmol, 1.0 eq) in N-methylpyrrolidone (34.0 ml) at 25 ℃ with stirring under nitrogen atmosphere was added N, N-diisopropylethylamine (3.2 g, 23.4 mmol, 2.0 eq). The resulting mixture was stirred at 90 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature, purified by reverse phase chromatography (C18), eluted with a mobile phase of 5% → 95% acetonitrile/water (0.1% trifluoroacetic acid) over 25 minutes, and a detector: UV 254/220 nm. Compound 11-1 was obtained (yellow-green oil, 1.4 g, 22% yield). MS (ESI, m/z) 514.4[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ8.87(s,1H),7.62(d,J＝8.3Hz,1H),7.02(d,J＝2.1Hz,1H),6.75–6.71(m,1H),5.16(s,1H),4.97–4.91(m,1H),3.52–3.44(m,4H),3.24(t,J＝6.3Hz,2H),2.98–2.59(m,3H),2.46(d,J＝5.5Hz,6H),2.21–2.09(m,1H),1.82–1.59(m,4H),1.48(s,9H)。

Step 2

To a solution of compound 11-1 (350.0 mg, crude) in dichloromethane (4.0 ml) was added trifluoroacetic acid (1.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave crude product 11-2 (yellow solid, 350 mg). The compound was used in the next synthesis without further purification. MS (ESI, m/z) 414.1[ 2 ], [ M ] +H] ⁺ 。

Step 3

Compound 11-2 (37.4 mg, 0.07 mmol, 1.2 equivalents), sodium acetate (6.2 mg, 0.07 mmol, 1.0 equivalent), acetic acid (4.5 mg, 0.07 mmol, 1.0 equivalent), sodium cyanoborohydride (9.4 mg, 0.14 mmol, 2.0 equivalents) and methanol (2.0 ml) were added to the reaction flask with stirring at 25 ℃, and then a solution of compound 10-4a (50.0 mg, 0.07 mmol, 1.0 equivalent) in methanol (0.5 ml) was added dropwise thereto. The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, giving compound 11-3a (yellow solid, 35 mg, yield 44%). MS (ESI, m/z) 1063.4/1065.4[ 2 ], [ M + H ] ] ⁺ 。

Step 4

To a solution of compound 11-3a (35.0 mg, 0.03 mmol, 1.0 eq) in dichloromethane (0.9 ml) was added trifluoroacetic acid (0.3 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 24% b phase gradient over 7 minutes; detector UV 254/220 nm; compound 11 was obtained (yellow solid, 15.0 mg, 44% yield). MS (ESI, m/z) 963.4/965.4[ m ] +H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ11.07(s,1H),8.18(s,1H),8.01(d,J＝1.6Hz,1H),7.82(d,J＝8.3Hz,1H),7.56(d,J＝8.3Hz,1H),7.47–7.42(m,1H),7.30(d,J＝2.3Hz,1H),7.26–7.16(m,2H),7.14–7.11(m,1H),7.07(d,J＝2.4Hz,1H),6.94(d,J＝2.0Hz,1H),6.86–6.82(m,1H),5.06–5.00(m,1H),4.43–4.38(m,2H),3.91–3.89(m,6H),3.17–3.15(m,7H),2.94–2.80(m,1H),2.75–2.64(m,2H),2.64–2.55(m,2H),2.41–2.22(m,14H),2.02–1.96(m,3H),1.53–1.45(m,8H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-73.48,-122.22。

Example 12

4- ((4- (4- ((3- ((4- ((1s, 4s) -2, 5-diazabicyclo [2.2.1] heptan-2-yl) -6-chloro-8-fluoro-7- ((S or R) -3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione formate 12a;4- ((4- (4- ((3- ((4- ((1s, 4s) -2, 5-diazabicyclo [2.2.1] heptan-2-yl) -6-chloro-8-fluoro-7- ((R or S) -3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione dicarboxylate 12b

Step 1

To a solution of compounds 1-6 (1.5 g, 4.3 mmol, 1.0 eq) in ultra dry 1, 4-dioxane (15.0 ml) was added triethylamine (1.8 ml, 13.0 mmol, 3.0 eq) at 0 ℃ with stirring under nitrogen, followed by dropwise addition of (1S, 4S) -2, 5-diazabicyclo [2.2.1 ]]A super dry 1, 4-dioxane (3.0 ml) solution of tert-butyl heptane-2-carboxylate (0.9 g, 4.3 mmol, 1.0 eq). The resulting mixture was reacted at 25 ℃ for 2 hours, the course of which was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 35% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 12-1 (yellow solid, 1.9 g, 85% yield). MS (ESI, m/z) 491.0/493.0/495.0M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.88(d,J＝11.6Hz,1H),5.47–5.33(m,1H),4.86–4.66(m,1H),4.18–4.15(m,1H),4.05–3.77(m,1H),3.60–3.56(m,2H),2.08–2.05(m,2H),1.45(s,9H)。

Step 2

To a solution of compound 12-1 (1.5 g, 2.3 mmol, 1.0 eq) in N-methylpyrrolidone (15.0 ml) was added cesium fluoride (0.9 g, 5.8 mmol, 2.0 eq), N-diisopropylethylamine (1.5 ml, 8.7 mmol, 3.0 eq) and compound 4-1 (960.0 mg, 5.8 mmol, 2.0 eq) at 25 ℃ with stirring under nitrogen. Adding the resulting mixture completely The reaction was stirred for 4 hours with the temperature increasing to 130 ℃ and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the reaction solution was cooled to 25 ℃. The reaction solution was directly purified by reverse phase flash chromatography (C18 column) and eluted with 50% → 95% methanol/water mobile phase (0.1% ammonia) within 20 minutes; detector UV254 nm; compound 12-2 was obtained (pale yellow solid, 750.0 mg, 42% yield). MS (ESI, m/z): 612.2/614.2/616.2M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.80(d,J＝11.6Hz,1H),5.88–5.75(m,1H),5.37–5.24(m,1H),5.07–4.90(m,2H),4.81–4.61(m,1H),4.50–4.46(m,2H),4.15–4.12(m,1H),3.99–3.47(m,3H),2.61–2.57(m,2H),2.48–2.36(m,2H),2.29(s,3H),2.14–2.03(m,6H),1.65–1.55(m,2H),1.47–1.44(m,9H)。

Step 3

To a solution of compound 12-2 (750.0 mg, 1.2 mmol, 1.0 eq) in tetrahydrofuran/water (10/1, 8.3 ml) was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (495.8 mg, 1.7 mmol, 1.5 eq), potassium phosphate (519.4 mg, 2.3 mmol, 2.0 eq), and chlorine (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl ] at 25 ℃ with stirring under nitrogen protection]Palladium (II) (96.2 mg, 0.1 mmol, 0.1 eq). The resulting mixture was stirred at 60 ℃ for 1.5 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, cooling the reaction liquid to 25 ℃, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and to obtain compound 12-3 (pale yellow solid, 673.0 mg, 86% yield). MS (ESI, m/z) 676.4/678.4[ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.88–7.85(m,1H),7.74–7.71(m,1H),7.42–7.38(m,1H),7.29(s,1H),7.28–7.16(m,2H),7.13–7.09(m,1H),5.83–5.69(m,1H),5.37–5.29(m,1H),5.05–4.87(m,2H),4.72–4.70(m,1H),4.48(s,2H),4.22–3.97(m,2H),3.80–3.78(m,1H),3.60–3.57(m,1H),2.69–2.64(m,2H),2.52–2.38(m,2H),2.31(s,3H),2.07–2.00(m,6H),1.67–1.57(m,2H),1.45(s,9H)。

Step 4

Chiral resolution of the compound 12-3 (673.0 mg) obtained in step 3 by preparative chiral high performance liquid chromatography: chiral column CHIRALPAK IA,2 × 25 cm, 5 microns; mobile phase A: n-hexane (10 mmol/l ammonia-methanol), mobile phase B: isopropyl alcohol; flow rate: 20 ml/min; elution was performed with 50% phase B over 22 minutes; detector UV 220/254 nm, resulting in two products. The product of shorter retention time (4.75 min) was compound 12-3a (light yellow solid, 330.0 mg, 49% recovery), compound 12-3a: MS (ESI, m/z) 676.4/678.4[ 2 ], [ M + H ]] ⁺ (ii) a The product at longer retention time (10.76 min) was compound 12-3b (pale yellow solid, 260.0 mg, 39% recovery), compound 12-3b: MS (ESI, m/z) 676.4/678.4[ M ] +H] ⁺ 。

Step 5

To a solution of compound 12-3a (330.0 mg, 0.5 mmol, 1.0 equiv) in acetone/water (4/1, 6.3 mL) was added N-methylmorpholine oxide (85.8 mg, 0.7 mmol, 1.5 equiv) and potassium osmate dihydrate (18.0 mg, 0.05 mmol, 0.1 equiv) sequentially with stirring at 0 deg.C. The resulting mixture was stirred at 0 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was directly purified by a reverse phase flash column (C18 column), and eluted with a methanol/water mobile phase (0.1% ammonia) of 50% → 95% in 20 minutes; detector UV254 nm; compound 12-4a (Ash) is obtained Colored solid, 184 mg, 56% yield). MS (ESI, m/z) 710.4/712.4[ m ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.84(s,1H),7.75–7.72(m,1H),7.43–7.38(m,1H),7.31(s,2H),7.25–7.16(m,2H),5.36–5.32(m,1H),4.78–4.66(m,1H),4.48–4.45(m,2H),4.19–4.16(m,1H),4.04–4.01(m,1H),3.88–3.73(m,1H),3.67–3.49(m,4H),3.44–3.38(m,1H),2.70–2.54(m,2H),2.42(s,2H),2.26(s,3H),2.08–2.03(m,4H),1.68–1.65(m,3H),1.49(s,9H)。

Step 6

To a solution of compound 12-4a (75.0 mg, 0.1 mmol, 1.0 eq) in acetonitrile/water (4/1,1.5 ml) was added sodium periodate (128.7 mg, 0.6 mmol, 6.0 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, water (10 ml), dichloromethane (10 ml × 3) was added to the reaction solution for extraction, the organic phases were combined, the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product 12-5a (pale yellow oily liquid, 80.0 mg) which was used directly in the next reaction. MS (ESI, m/z) 678.3/680.3[ M + H ]] ⁺ 。

Step 7

To a reaction flask were added compounds 1 to 4 (61.6 mg, 0.14 mmol, 1.2 equivalents), sodium acetate (10.2 mg, 0.12 mmol, 1.0 equivalent), acetic acid (7.5 mg, 0.12 mmol, 1.0 equivalent), sodium cyanoborohydride (15.6 mg, 0.24 mmol, 2.0 equivalents) and methanol (2.0 ml) in this order under stirring at 25 ℃, and then a solution of compound 12 to 5a (80.0 mg, 0.12 mmol, 1.0 equivalent) in methanol (0.5 ml) was added dropwise. The obtained mixture is continuously stirred and reacted for 2 hours at the temperature of 25 ℃ and reacted The course is monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, giving compound 12-6a (yellow solid, 46 mg, yield 45%). MS (ESI, m/z) 1075.4/1077.4[ 2 ], [ M ] +H] ⁺ 。

Step 8

To a solution of compound 12-6a (46.0 mg, 0.04 mmol, 1.0 eq) in dichloromethane (2.0 ml) was added trifluoroacetic acid (0.5 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 10% → 95% methanol/water mobile phase (0.1% formic acid) over 25 minutes; detector UV254 nm; compound 12a was obtained (yellow solid, 20 mg, 47% yield). MS (ESI, m/z) 975.4/977.4[ m + H ]] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ11.11(s,1H),8.19(s,1H),8.05(d,J＝1.7Hz,1H),7.82(d,J＝8.3Hz,1H),7.57–7.55(m,1H),7.46–7.44(m,1H),7.30(d,J＝2.4Hz,1H),7.24–7.21(m,1H),7.18–7.07(m,3H),7.02(d,J＝7.0Hz,1H),6.57–6.53(m,1H),5.19(s,1H),5.06–5.02(m,1H),4.47–4.32(m,4H),4.27(s,2H),3.99(d,J＝10.0Hz,2H),3.44–3.19(m,5H),2.92–2.80(m,1H),2.70–2.56(m,4H),2.37–2.34(m,6H),2.30–2.24(m,8H),2.12–2.10(m,1H),2.08–1.87(m,4H),1.50–1.44(m,8H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-122.09。

Step 5'

To compound 12-3b (260.0 mg) was added with stirring at 0 deg.C0.4 mmol, 1.0 equiv) in acetone/water (4/1, 5.0 mL) was added N-methylmorpholine oxide (67.6 mg, 0.5 mmol, 1.5 equiv) and potassium osmate dihydrate (14.2 mg, 0.04 mmol, 0.1 equiv) sequentially. The resulting mixture was stirred at 0 ℃ for an additional 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was directly purified by a reverse phase flash column (C18 column), and eluted with a methanol/water mobile phase (0.1% ammonia) of 50% → 95% in 20 minutes; detector UV254 nm; compound 12-4b was obtained (grey solid, 200.0 mg, 77% yield). MS (ESI, m/z) 710.4/712.4[ m ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.84(s,1H),7.75–7.72(m,1H),7.43–7.38(m,1H),7.31(s,2H),7.25–7.16(m,2H),5.36–5.32(m,1H),4.78–4.66(m,1H),4.48–4.45(m,2H),4.19–4.16(m,1H),4.04–4.01(m,1H),3.88–3.73(m,1H),3.67–3.49(m,4H),3.44–3.38(m,1H),2.70–2.54(m,2H),2.42(s,2H),2.26(s,3H),2.08–2.03(m,4H),1.68–1.65(m,3H),1.49(s,9H)。

Step 6'

To a solution of compound 12-4b (75.0 mg, 0.1 mmol, 1.0 eq) in acetonitrile/water (4/1,1.5 ml) was added sodium periodate (128.7 mg, 0.6 mmol, 6.0 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, water (10.0 ml), dichloromethane (10.0 ml × 3) were added to the reaction solution for extraction, the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product 12-5b (pale yellow oily liquid, 80.0 mg) which was used directly in the next reaction. MS (ESI, m/z) 678.4/680.4[ M ] +H] ⁺ 。

Step 7'

To a reaction flask were added compounds 1-4 (65.5 mg, 0.12 mmol, 1.2 equivalents), sodium acetate (10.8 mg, 0.13 mmol, 1.0 equivalent), acetic acid (7.9 mg, 0.13 mmol, 1.0 equivalent), sodium cyanoborohydride (16.6 mg, 0.25 mmol, 2.0 equivalents) and methanol (2.0 ml) in this order with stirring at 25 ℃, and then 12-5b (85.0 mg, 0.13 mmol, 1.0 equivalent) in methanol (0.5 ml) was added dropwise. The resulting mixture was stirred at 25 ℃ for an additional 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 12-6b (yellow solid, 56 mg, yield 49%) was obtained. MS (ESI, m/z) 1075.4/1077.4[ 2 ], [ M ] +H ] ⁺ 。

Step 8'

To a solution of compound 12-6b (56.0 mg, 0.05 mmol, 1.0 eq) in dichloromethane (2.0 ml) was added trifluoroacetic acid (0.5 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 10% → 95% methanol/water mobile phase (0.1% formic acid) over 20 minutes; detector UV254 nm; compound 12b was obtained (yellow solid, 30 mg, 57% yield). MS (ESI, m/z) 975.4/977.4[ m + H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.10(s,1H),8.19(s,2H),8.07–8.02(m,1H),7.81(d,J＝8.3Hz,1H),7.58–7.54(m,1H),7.46–7.41(m,1H),7.29(d,J＝2.4Hz,1H),7.24–7.13(m,2H),7.10(d,J＝8.6Hz,1H),7.06–6.98(m,2H),6.55(t,J＝5.9Hz,1H),5.20–5.17(m,1H),5.08–5.02(m,1H),4.38–4.32(m,4H),4.16(s,2H),3.93(d,J＝9.9Hz,2H),3.31–3.25(m,3H),3.22–3.20(m,1H),2.94–2.83(m,1H),2.62–2.60(m,1H),2.58–2.55(m,3H),2.45–2.42(m,3H),2.39–2.31(m,4H),2.29–2.21(m,8H),2.10–2.00(m,2H),2.09–2.01(m,3H),1.61–1.50(m,2H),1.49–1.36(m,6H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-122.02。

Example 13

4- ((2- (2- (4- (2- (2- ((3- ((S or R) -4- ((1r, 5s) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) ethoxy) ethyl) piperazin-1-yl) ethoxy) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione pentatrifluoroacetate 13a;4- ((2- (2- (4- (2- (2- ((3- ((R or S) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) ethoxy) ethyl) piperazin-1-yl) ethoxy) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione pentatrifluoroacetate 13b.

Step 1

To a 250 ml round bottom flask was added potassium phthalimide salt (5.0 g, 25.6 mmol, 1.0 eq), acetone (50.0 ml) and 2,2' -dibromodiethyl ether (11.9 g, 51.3 mmol, 2.0 eq) with stirring at 25 degrees celsius. The resulting mixture was warmed to 60 degrees celsius and stirred at that temperature for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is cooled to room temperature, and a crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 10% → 30% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 13-1 (off-white solid, 6.4 g, 84% yield). MS (ESI, m/z) 298.0/300.0[ 2 ] M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.90–7.82(m,2H),7.76–7.69(m,2H),3.94-3.90(m,2H),3.82-3.76(m,4H),3.41(t,J＝6.2Hz,2H)。

Step 2

To a 500 ml round bottom flask was added compound 13-1 (6.4 g, 20.4 mmol, 1.0 eq), 1-tert-butoxycarbonylpiperazine (4.2 g, 22.4 mmol, 1.1 eq), acetonitrile (100.0 ml) and N, N-diisopropylethylamine (7.1 ml, 40.8 mmol, 2.0 eq) with stirring at 25 ℃. The mixture was warmed to 80 ℃ and stirred at this temperature for 4 hours, the course of the reaction being monitored by liquid and thin layer chromatography. After the reaction is finished, cooling the reaction liquid to room temperature, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 13-2 (dark yellow oily liquid, 7.0 g, 85% yield). MS (ESI, m/z) 404.3[ 2 ], [ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.87–7.81(m,2H),7.76–7.68(m,2H),3.90(t,J＝5.7Hz,2H),3.71(t,J＝5.6Hz,2H),3.61(t,J＝5.5Hz,2H),3.41–3.29(m,4H),2.53(t,J＝5.5Hz,2H),2.37(t,J＝5.1Hz,4H),1.45(s,9H)。

Step 3

To a solution of compound 13-2 (7.0 g, 16.5 mmol, 1.0 eq) in ethanol (300.0 ml) was added hydrazine hydrate (4.1 g, 66.0 mmol, 4.0 eq) with stirring at 25 ℃. The resulting mixture was stirred at 80 ℃ for 16 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the reaction was cooled to room temperature, the insoluble material was removed by filtration, the filter cake was washed with ethanol (20 ml x 2), and the filtrate was concentrated under reduced pressure to give a crude product. Dichloro-methane for crude productThe alkane (300 ml) was dissolved, the insolubles were removed by filtration, the filter cake was washed with dichloromethane (20 ml x 2) and the filtrate was concentrated under reduced pressure to give crude 13-3 (a pale yellow oily liquid, 4.1 g). The compound was used in the next synthesis without further purification. MS (ESI, m/z) 274.2[ 2 ], [ M + H ]] ⁺ 。

Step 4

To a 50 ml three-necked flask, compound 13-3 (2.0 g, 7.0 mmol, 1.0 eq), N-methylpyrrolidone (20.0 ml), N-diisopropylethylamine (1.81 g, 14 mmol, 2 eq) and 2- (2, 6-dioxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (2.2 g, 7.6 mmol, 1.1 eq) were added at 25 degrees celsius with stirring under nitrogen. The resulting mixture was stirred at 90 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was purified by reverse phase chromatography (C18), eluted with a mobile phase of 10% → 95% methanol/water (0.1% trifluoroacetic acid) within 30 minutes, with a detector UV 254/220 nm; compound 13-4 was obtained (yellow solid, 1.4 g, 38% yield). MS (ESI, m/z) 530.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ8.94(s,1H),7.54–7.48(m,1H),7.14–7.12(m,1H),6.92–6.89(m,1H),6.54–6.51(m,1H),4.97–4.84(m,1H),3.72–3.68(m,4H),3.55–3.41(m,6H),2.94–2.61(m,5H),2.53–2.49(m,4H),2.22–2.12(m,1H),1.48(s,9H)。

Step 5

To a solution of compound 13-4 (500.0 mg, 0.9 mmol, 1.0 eq) in dichloromethane (8.0 ml) was added trifluoroacetic acid (2.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, decreaseConcentration under pressure gave crude 13-5 (yellow solid, 740.0 mg). The compound was used directly in the subsequent synthesis without further purification. MS (ESI, m/z) 430.2[ 2 ], [ M ] +H] ⁺ 。

Step 6

To a solution of compound 2- (allyloxy) ethan-1-ol (8.0 g, 77.5 mmol, 1.0 equiv) and pyridine (18.6 g, 232.6 mmol, 3.0 equiv) in dichloromethane (50.0 ml) was added p-toluenesulfonyl chloride (22.4 g, 116.3 mmol, 1.5 equiv) in portions with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 30% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and to obtain compound 13-6 (colorless liquid, 20.0 g, yield 99%). MS (ESI, m/z) 257.3[ m + H ] ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.86–7.80(m,2H),7.39–7.33(m,2H),5.91–5.78(m,1H),5.35–5.15(m,2H),4.24–4.16(m,2H),3.98–3.95(m,2H),3.69–3.62(m,2H),2.47(s,3H)。

Step 7

To a solution of compound 13-6 (19.0 g, 73.4 mmol, 1.0 eq) in acetonitrile (100.0 ml) was added potassium carbonate (30.7 g, 220.15 mmol, 3.0 eq) and 3- (methylamino) -1-propanol (85.9 g, 95.4 mmol, 1.3 eq) with stirring at 25 ℃. The resulting mixture was stirred at 85 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product with silica gel column chromatography, eluting with 0% → 30% ethyl acetate/petroleum ether mobile phase gradient, and collecting the fractionsThe solvent was removed by rotary evaporation under reduced pressure to give compound 13-7 (pale yellow oily liquid, 11.0 g, yield 86%). MS (ESI, m/z) 174.2[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ5.99–5.87(m,1H),5.34–5.17(m,2H),4.02–3.99(m,2H),3.85–3.77(m,2H),3.58(t,J＝5.8Hz,2H),2.70–2.64(m,4H),2.34(s,3H),1.78–1.67(m,2H).

Step 8

To a solution of compound 3-2 (1.5 g, 2.8 mmol, 1.0 eq), cesium fluoride (0.9 g, 5.6 mmol, 2.0 eq) and N, N-diisopropylethylamine (1.6 ml, 8.4 mmol, 3.0 eq) in N-methylpyrrolidinone (15.0 ml) at 25 ℃ with stirring under nitrogen protection was added compound 13-7 (1.0 g, 5.6 mmol, 2.0 eq). The resulting mixture was stirred at 130 ℃ for 4 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature. The reaction solution was directly purified by reverse phase flash chromatography (C18 column) and eluted with 50% → 95% methanol/water mobile phase (0.1% ammonia) over 25 minutes; detector UV254 nm; compound 13-8 was obtained (yellow oily liquid, 1.1 g, 61% yield). MS (ESI, m/z) 642.2/644.2/646.2M + H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.70(d,J＝2.0Hz,1H),5.98–5.85(m,1H),5.33–5.11(m,2H),4.48(t,J＝6.6Hz,2H),4.33–4.29(m,4H),4.01–3.99(m,2H),3.57–3.53(m,4H),2.65–2.61(m,4H),2.32(s,3H),2.09–1.94(m,4H),1.85–1.81(m,2H),1.53(s,9H).

Step 9

To a solution of compound 13-8 (1.2 g, 1.8 mmol, 1.0 eq.) in tetrahydrofuran/water (10/1, 13.2 ml) was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaboroza at 25 ℃ with stirring under nitrogen protectionCyclopentan-2-yl) naphthalen-2-ol (0.7 g, 2.7 mmol, 1.5 equivalents), potassium phosphate (0.8 g, 3.5 mmol, 2.0 equivalents) and chlorine (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl)]Palladium (II) (139.3 mg, 0.2 mmol, 0.1 eq). The resulting mixture was stirred at 60 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, cooling the reaction liquid to room temperature, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 13-9 (pale yellow solid, 710.0 mg, 57% yield). MS (ESI, m/z) 706.4/708.4[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.78–7.71(m,2H),7.44–7.39(m,1H),7.34–7.31(m,2H),7.24–7.21(m,1H),7.18–7.15(m,1H),5.92–5.79(m,1H),5.26–5.10(m,2H),4.53–4.33(m,6H),3.96–3.93(m,2H),3.65–3.62(m,4H),2.83–2.81(m,4H),2.47(s,3H),2.23–2.09(m,2H),1.99–1.97(m,2H),1.85–1.83(m,2H),1.55(s,9H)。

Step 10

Chiral resolution of the compound 13-9 (710.0 mg) obtained in step 9 by preparative chiral high performance liquid chromatography: a chiral column NB-Lux 5 mi-Cellulose-5, 2.12x25 cm, 5 microns; a mobile phase A: n-hexane/dichloromethane =5:1 (0.5% 2 mol/l ammonia-methanol), mobile phase B: ethanol; flow rate: 20 ml/min; elution was performed with 20% phase B over 9 minutes, detector UV 220/254 nm. Two products were obtained, a shorter retention time (5 minutes) of the product being compound 13-9a (pale yellow solid, 310.0 mg, 44% recovery), compound 13-9a: MS (ESI, m/z) 706.4/708.4[ 2 ], [ M + H ] ] ⁺ (ii) a The product at longer retention time (7.2 min) was compound 13-9b (light yellow solid, 300.0 mg, 42% recovery), compound 13-9b: MS (ESI, m/z) 706.4/708.4[ m + H ]] ⁺ 。

Step 11

To a solution of compounds 13-9a (300.0 mg, 0.4 mmol, 1.0 eq) in acetone/water (4/1, 3.8 mL) was added N-methylmorpholine oxide (70.9 mg, 0.6 mmol, 1.5 eq) and potassium osmate dihydrate (14.9 mg, 0.04 mmol, 0.1 eq) with zero degrees Centigrade stirring. The resulting mixture was stirred at zero degrees centigrade for 4 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was directly purified by a reverse phase flash chromatography column (C18 column), and eluted with a methanol/water mobile phase (0.1% ammonia water) of 50% → 95% in 25 minutes; detector UV254 nm; compound 13-10a was obtained (brown solid, 160.0 mg, 54% yield). MS (ESI, m/z) 740.4/742.4[ 2 ] M + H] ⁺ 。

Step 12

To a solution of compound 13-10a (60.0 mg, 0.1 mmol, 1.0 equiv) in acetonitrile/water (4/1, 2.5 ml) was added sodium periodate (104.0 mg, 0.5 mmol, 6.0 equiv) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, water (10.0 ml) was added to dilute the reaction, the mixture was extracted with dichloromethane (10 ml x 3), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give crude product 13-11a (pale yellow oily liquid, 65.0 mg) which was used directly in the next reaction. MS (ESI, m/z) 708.4/710.4[ 2 ] M + H ] ⁺ 。

Step 13

To a solution of compound 13-5 (47.3 mg, 0.11 mmol, 1.2 equiv), sodium acetate (7.5 mg, 0.09 mmol, 1.0 equiv), acetic acid (0.01 ml, 0.09 mmol, 1.0 equiv) and sodium cyanoborohydride (11.5 mg, 0.18 mmol, 2.0 equiv) in methanol (2.0 ml) was added compound 13-11a (65.0 mg, 0.09 mmol, 1.0 equiv) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, giving compound 13-12a (yellow solid, 40 mg, yield 42%). MS (ESI, m/z) 1121.4/1123.4[ M ] +H] ⁺ 。

Step 14

To a solution of compound 13-12a (40.0 mg, 0.03 mmol, 1.0 eq) in dichloromethane (2.0 ml) was added trifluoroacetic acid (0.5 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 33% B phase gradient over 7 minutes; detector UV 254/220 nm; compound 13a was obtained (yellow solid, 32.0 mg, 59% yield). MS (ESI, m/z) 1021.4/1023.4[ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.01(d,J＝1.6Hz,1H),7.76(d,J＝8.3Hz,1H),7.58–7.54(m,1H),7.46–7.40(m,1H),7.31–7.17(m,3H),7.13–7.01(m,3H),5.12–4.99(m,1H),4.72(d,J＝14.3Hz,2H),4.63–4.57(m,2H),4.27(s,2H),3.99–3.82(m,4H),3.81–3.61(m,6H),3.51–3.37(m,10H),3.28–3.05(m,8H),2.99(s,3H),2.88–2.63(m,3H),2.38–2.29(m,2H),2.21–2.04(m,5H)； ¹⁹ F NMR(282MHz,CD ₃ OD)δ-77.08,-122.94,-122.97。

Step 11'

To a solution of compounds 13-9b (300.0 mg, 0.4 mmol, 1.0 eq) in acetone/water (4/1, 5.0 mL) was added N-methylmorpholine oxide (70.9 mg, 0.6 mmol, 1.5 eq) and potassium osmate dihydrate (14.9 mg, 0.04 mmol, 0.1 eq) with zero degrees Centigrade stirring. The resulting mixture was stirred at zero degrees centigrade for 4 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was directly purified by reverse phase flash chromatography (C18 column) and eluted with 50 → 95% methanol/water mobile phase (0.1% ammonia) in 25 minutes; detector, UV254 nm; compound 13-10b was obtained (brown solid, 136.0 mg, 45% yield). MS (ESI, m/z) 740.4/742.4[ M + H ]] ⁺ 。

Step 12'

To a solution of compound 13-10b (60.0 mg, 0.1 mmol, 1.0 eq) in acetonitrile/water (4/1,2.5 ml) was added sodium periodate (104.0 mg, 0.5 mmol, 6.0 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, water (10.0 ml) was added to dilute the reaction, the mixture was extracted with dichloromethane (10 ml x 3), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give crude product 13-11b (pale yellow oily liquid, 62.0 mg) which was used directly in the next reaction. MS (ESI, m/z) 708.2/710.2[ 2 ], [ M + H ] ] ⁺ 。

Step 13'

To a solution of compound 13-5 (45.1 mg, 0.10 mmol, 1.2 equiv.), sodium acetate (7.2 mg, 0.08 mmol, 1.0 equiv.), acetic acid (5.3 mg, 0.08 mmol, 1.0 equiv.) and sodium cyanoborohydride (11.0 mg, 0.17 mmol, 2.0 equiv.) in methanol (2.0 ml) was added compound 13-11b (62.0 mg, 0.08 mmol, 1.0 equiv.) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 13-12b (yellow solid, 45.0 mg, yield 45%) was obtained. MS (ESI, m/z) 1121.4/1123.4[ M ] +H] ⁺ 。

Step 14'

To a solution of compound 13-12b (45.0 mg, 0.04 mmol, 1.0 eq) in dichloromethane (2.0 ml) was added trifluoroacetic acid (0.5 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 33% b phase gradient over 7 minutes; detector UV 254/220 nm; compound 13b was obtained (yellow solid, 33.0 mg, 54% yield). MS (ESI, m/z) 1021.4/1023.4[ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.08(d,J＝1.7Hz,1H),7.83(d,J＝8.3Hz,1H),7.65–7.60(m,1H),7.52–7.47(m,1H),7.37–7.23(m,3H),7.17–7.09(m,3H),5.18–5.07(m,1H),4.79(d,J＝14.2Hz,2H),4.67(t,J＝5.8Hz,2H),4.34(s,2H),4.04–3.89(m,4H),3.87–3.82(m,4H),3.78–3.73(m,2H),3.65–3.50(m,6H),3.45(s,3H),3.36–3.22(m,6H),3.16(s,2H),3.06(s,3H),2.98–2.67(m,4H),2.44–2.33(m,2H),2.24(s,4H),2.20–2.09(m,1H)； ¹⁹ F NMR(282MHz,CD ₃ OD)δ-77.09,-122.95,-122.98。

Example 14

4- ((5- (1- (5- ((3- ((S or R) -4- ((1r, 5s) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) pentyl) piperidin-4-yl) pentyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione trihydrochloride 14.

Step 1

To a solution of compound 4- (3-hydroxypropyl) piperidine-1-carboxylic acid tert-butyl ester (15.0 g, 58.6 mmol, 1.0 eq) in dichloromethane (300.0 ml) was added pyridinium chlorochromate (25.8 g, 117.1 mmol, 2.0 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the insoluble material was removed by filtration, the filter cake was washed with dichloromethane (2 × 20 ml), and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography on silica gel eluting with a 0% → 50% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 14-1 (pale yellow oil, 6.0 g, 38% yield). MS (ESI, m/z) 242.3[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ9.68(t,J＝1.6Hz,1H),3.92(d,J＝13.1Hz,2H),2.65(s,2H),2.47–2.43(m,2H),1.66–1.56(m,2H),1.51–1.43(m,2H),1.39(s,9H),1.35–1.30(m,1H),1.02–0.86(m,2H).

Step 2

To a solution of diethyl cyanomethylphosphonate (4.4 g, 23.6 mmol, 1.2 equivalents) in tetrahydrofuran (6 ml) was added dropwise lithium bistrimethylsilyl amide (1 mol/l in tetrahydrofuran, 25.2 ml, 25.2 mmol, 1.2 equivalents) and a solution of compound 14-1 (5.5 g, 20.5 mmol, 1.0 equivalents) in tetrahydrofuran (2.0 ml) at-78 deg.c with stirring under nitrogen protection. The resulting mixture was stirred at-78 ℃ for 40 minutes and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, adding a saturated ammonium chloride solution (30 ml) into the reaction solution at zero centigrade to quench the reaction, then extracting the reaction system with dichloromethane (50 ml x 3), combining organic phases, drying the organic phases with anhydrous sodium sulfate, filtering to remove a drying agent, and concentrating the filtrate to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 30% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 14-2 (pale yellow liquid, 3.8 g, 67% yield). MS (ESI, m/z) 265.2[ 2 ], [ M ] +H ] ⁺ 。

Step 3

To a solution of compound 14-2 (3.8 g, 13.7 mmol, 1.0 eq) in methanol (35.0 ml) was added raney nickel (380.0 mg) and ammonia in methanol (7 mol/l, 10 ml) at 25 ℃ with stirring under nitrogen. The resulting mixture was stirred at 25 ℃ for 16 hours under 1 atmosphere of hydrogen and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, filtering by using kieselguhr to remove the nickel catalyst, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product is purified by silica gel column chromatography with 0% → 10% methanol-The dichloromethane mobile phase was subjected to gradient elution, and the obtained fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, thereby obtaining compound 14-3 (yellow liquid, 2.2 g, yield 57%). MS (ESI, m/z) 271.2, M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.08(d,J＝13.0Hz,2H),2.76–2.60(m,4H),1.65(d,J＝13.3Hz,2H),1.51–1.42(m,12H),1.37–1.30(m,6H),1.29–1.21(m,2H),1.15–1.02(m,2H)。

Step 4

To a 50 ml three-necked flask, compound 14-3 (1.1 g, 3.9 mmol, 1.1 eq), N-methylpyrrolidone (8.0 ml), N-diisopropylethylamine (928.8 mg, 7.2 mmol, 2.0 eq) and 2- (2, 6-dioxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (1.0 g, 3.6 mmol, 1.0 eq) were added at 25 degrees celsius with stirring under nitrogen. The mixture was stirred at 90 ℃ for 3 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was purified by reverse phase flash chromatography (C18) eluting with a mobile phase of acetonitrile/water (0.1% trifluoroacetic acid) 10% → 95% in 30 minutes with a detector UV 254/220 nm; compound 14-4 was obtained (yellow solid, 647.0 mg, 34% yield). MS (ESI, m/z) 527.2[ 2 ], [ M ] +H ] ⁺ 。

Step 5

To a solution of compound 14-4 (300.0 mg, 0.57 mmol, 1.0 eq) in dichloromethane (9.0 ml) was added trifluoroacetic acid (3.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave crude 14-5 (yellow solid, 364.0 mg). The crude product was used directly in the next synthesis. MS (ESI, m/z) 427.2[ 2 ], [ M ] +H] ⁺ 。

Step 6

To a solution of compound 14-5 (30.9 mg, 0.07 mmol, 1.2 equiv.), sodium acetate (7.4 mg, 0.09 mmol, 1.5 equiv.), acetic acid (5.2 mg, 0.09 mmol, 1.5 equiv.) and sodium cyanoborohydride (11.4 mg, 0.17 mmol, 3.0 equiv.) in methanol (2.0 ml) was added dropwise a solution of compound 3-6a (45.0 mg, 0.06 mmol, 1.0 equiv.) in methanol (0.5 ml) with stirring at 25 ℃. The resulting mixture was stirred at 50 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography, eluted with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the obtained fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 14-6a (yellow solid, 40 mg, yield 59%) was obtained. MS (ESI, m/z) 1116.6/1118.6[ M + H ] ] ⁺ 。

Step 7

To a solution of compound 14-6a (40.0 mg, 0.036 mmol, 1.0 eq) in methanol (1.5 ml) was added a solution of 1, 4-dioxane (4 mol/l, 1.5 ml) of hydrochloric acid with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X150 mm, 5 μm; mobile phase A: water (0.1% hydrochloric acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: eluting with 5% → 35% B phase gradient in 7 min; a detector: UV 254/220 nm; compound 14 was obtained (yellow solid, 8.0 mg, 19% yield). MS (ESI, m/z) 1016.4/1018.4[M+H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ Heavy water exchange) δ 7.99 (s, 1H), 7.82 (d, J =8.3hz, 1h), 7.59 (m, 1H), 7.46 (m, 1H), 7.31 (d, J =2.4hz, 1h), 7.27-7.17 (m, 2H), 7.12-7.01 (m, 3H), 5.07-4.98 (m, 1H), 4.58-4.39 (m, 4H), 4.19 (s, 2H), 3.87 (t, J =13.0h, 2h), 3.40 (s, 1H), 3.33-3.27 (m, 3H), 3.23-3.01 (m, 3H), 2.99-2.93 (m, 2H), 2.89-2.80 (m, 2H), 2.78 (s, 3H), 2.62 (s, 1H), 2.24-2.18 (m, 2H), 2.11-1.88 (m, 6H), 1.86-1.76 (m, 2H), 1.73-1.65 (m, 4H), 1.60-1.56 (m, 3H), 1.48-1.15 (m, 12H); ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-122.04。

Example 15

4- ((4- (4- (4- ((3- ((4- ((1r, 5s) -3, 8-diazabicyclo [3.2.1] octan-8-yl) -6-chloro-8-fluoro-7- ((S or R) -3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione tetratrifluoroacetate 15a 4- ((4- (4- ((3- ((4- ((1r, 5s) -3, 8-diazabicyclo [3.2.1] octan-8-yl) -6-chloro-8-fluoro-7- ((R or S) -3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1-dione tetraacetate.

Step 1

To a solution of compounds 1-6 (1.50 g, 4.313 mmol, 1.00 eq) and triethylamine (1.31 g, 12.940 mmol, 3.00 eq) in 1, 4-dioxane (15.0 ml) was added dropwise 3, 8-diazabicyclo [3.2.1] at zero degrees centigrade, with stirring under nitrogen protection]Octane-3-carboxylic acid tert-butyl ester (0.92 g, 4.313 mmol, 1.00 eq). The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Silica gel column chromatography of crude productPurification, eluting with a 0% → 28% ethyl acetate/dichloromethane mobile phase gradient, and removal of solvent from the resulting fraction by rotary evaporation under reduced pressure gave compound 15-1 (off-white solid, 2.2 g, 91% yield). MS (ESI, m/z) 505.0/507.0/509.0[ 2 ], [ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.78(d,J＝2.0Hz,1H),4.99–4.84(m,2H),4.18–3.90(m,2H),3.47–3.18(m,2H),2.13–1.82(m,4H),1.49(s,9H)。

Step 2

To a solution of compound 15-1 (2.10 g, 3.941 mmol, 1.00 eq) in N-methylpyrrolidone (42 ml) was added cesium fluoride (1.26 g, 7.882 mmol, 2.0 eq), 4-1 (0.98 g, 5.912 mmol, 1.5 eq) and N, N-diisopropylethylamine (1.61 g, 11.823 mmol, 3.0 eq) at 25 degrees celsius with stirring under nitrogen. The resulting mixture was stirred at 130 ℃ for 6 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature. The reaction solution was directly purified by reverse phase flash chromatography (C18 column) and eluted with 5% → 95% methanol/water mobile phase (0.1% ammonium bicarbonate) within 30 minutes; detector, UV254 nm; compound 15-2 was obtained (colorless oil, 960 mg, 38% yield). MS (ESI, m/z) 626.2/628.2/630.2M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.73(d,J＝2.3Hz,1H),5.86–5.73(m,1H),5.06–4.91(m,2H),4.90–4.75(m,2H),4.54–4.43(m,2H),4.08–3.86(m,2H),3.47–3.20(m,2H),2.73–2.57(m,2H),2.49–2.39(m,2H),2.31(s,3H),2.10–2.02(m,4H),2.01–1.93(m,2H),1.90–1.80(m,2H),1.66–1.56(m,2H),1.49(s,9H)。

Step 3

Compound 15-2 (860.00 mg, 1) was added at 25 ℃ with stirring under nitrogen atmosphere372 mmol, 1.00 eq) of a tetrahydrofuran/water (10/1, 8.8 ml) mixed solution was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (555.79 mg, 2.058 mmol, 1.50 eq), potassium phosphate (582.29 mg, 2.744 mmol, 2.00 eq) and chlorine (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl) ]Palladium (II) (107.78 mg, 0.137 mmol, 0.10 eq). The reaction solution was stirred at 130 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, cooling the reaction liquid to 25 ℃, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 15-3 (off-white solid, 840 mg, 88% yield). MS (ESI, m/z) 690.4/692.3[ 2 ] M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.81(d,J＝1.6Hz,1H),7.76(d,J＝8.1Hz,1H),7.47–7.40(m,1H),7.34–7.31(m,1H),7.30–7.28(m,1H),7.27–7.19(m,1H),7.14(d,J＝2.4Hz,1H),5.85–5.68(m,1H),5.04–4.88(m,4H),4.55–4.45(m,2H),4.15–3.91(m,2H),3.50–3.28(m,2H),2.78–2.64(m,2H),2.58–2.44(m,2H),2.35(s,3H),2.16–2.00(m,6H),1.97–1.86(m,2H),1.69–1.59(m,2H),1.56–1.50(s,9H)。

Step 4

Chiral resolution of the compound 15-3 (840 mg) obtained in step 3 by preparative chiral high performance liquid chromatography: a chiral column NB-Lux 5 mi-Cellulose-5, 2.12x25 cm, 5 microns; mobile phase A: n-hexane (10 mmol/l ammonia), mobile phase B: isopropyl alcohol; flow rate: 18 ml/min; elution was performed with 50% of phase B in 17 minutes, detector UV 230/210 nm. Two products were obtained, a shorter retention time (6.0 minutes) product being compound 15-3a (pink solid, 235 mg, 29% recovery), compound 15-3a: MS (ESI, m/z) 690.4/692.3[ M + H ]] ⁺ (ii) a Of longer retention time (13.2 minutes) The product was compound 15-3b (pink solid, 300 mg, 37% recovery), compound 15-3b: MS (ESI, m/z) 690.4/692.3[ M + H ]] ⁺ 。

Step 5

To a solution of compound 15-3a (235.00 mg, 0.340 mmol, 1.00 equiv) in water/tetrahydrofuran (1/4, 5 ml) was added N-methylmorpholine oxide (59.82 mg, 0.511 mmol, 1.50 equiv) and potassium osmate dihydrate (50.18 mg, 0.136 mmol, 0.40 equiv) with stirring at 0 deg.C. The mixture was reacted at 0 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonia) over 30 minutes; detector UV254 nm; compound 15-4a was obtained (pink solid, 170 mg, 68% yield).

MS(ESI,m/z):724.4/726.3[M+H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.79(d,J＝1.7Hz,1H),7.73(d,J＝8.3Hz,1H),7.45–7.36(m,1H),7.33–7.27(m,2H),7.24–7.17(m,1H),7.15–7.10(m,1H),5.00–4.84(m,2H),4.58–4.40(m,2H),4.10–4.01(m,1H),3.98–3.89(m,1H),3.64–3.48(m,2H),3.43–3.23(m,3H),2.78–2.53(m,2H),2.50–2.38(m,2H),2.31–2.23(m,3H),2.12–1.81(m,7H),1.73–1.57(m,3H),1.50(s,9H)。

Step 6

To a mixed solution of compound 15-4a (70.00 mg, 0.092 mmol, 1.00 equiv) in water/tetrahydrofuran (1/4,5 ml) was added sodium periodate (157.81 mg, 0.580 mmol, 6.00 equiv) with stirring at zero degrees centigrade. The mixture is reacted for 3 hours at zero degrees centigrade, and the reaction process is carried out through a liquid substance and a thin layer Chromatography was used for monitoring. After completion of the reaction, the reaction solution was diluted with water (50 ml) and extracted with dichloromethane (50 ml × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude compound 15-5a as a white-like solid (61 mg). The crude product was used directly in the next reaction without further purification. MS (ESI, m/z) 692.3/694.3[ M ] +H] ⁺ 。

Step 7

To a solution of compound 15-5a (61 mg, 0.084 mmol, 1.00 eq), acetic acid (5.28 mg, 0.088 mmol, 1.05 eq) and sodium cyanoborohydride (11.10 mg, 0.177 mmol, 2.11 eq) in methanol (2 ml) was added dropwise a solution of compound 1-4 (30.97 mg, 0.071 mmol, 0.85 eq) in methanol (1 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonia) over 30 minutes; detector UV254 nm; compound 15-6a was obtained (yellow solid, 61 mg, 62% yield). MS (ESI, M/z) 1089.6/1091.6.[ M + H ] ] ⁺ 。

Step 8

To a solution of compound 15-6a (61.00 mg, 0.053 mmol, 1.00 eq) in dichloromethane (3 ml) was added trifluoroacetic acid (1 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μmRice; a mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: methanol; flow rate: 25 ml/min; gradient eluted with 25% → 54% b phase gradient in 7 min; a detector: UV 254/220 nm; yield 15a (light yellow solid, 25 mg, 31% yield). MS (ESI, m/z) 989.6/991.5[ m + H ]] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.06(d,J＝1.6Hz,1H),7.76(d,J＝8.3Hz,1H),7.58–7.52(m,1H),7.45–7.38(m,1H),7.28(d,J＝2.4Hz,1H),7.25–7.16(m,2H),7.10–7.02(m,3H),5.22–5.14(m,2H),5.09–5.01(m,1H),4.66–4.55(m,2H),3.71(d,J＝12.8Hz,2H),3.49–3.42(m,3H),3.37(t,J＝6.4Hz,3H),3.30–3.20(m,7H),3.13–2.98(m,5H),2.92(s,3H),2.88–2.78(m,3H),2.77–2.66(m,2H),2.41–2.26(m,4H),2.21–2.05(m,3H),1.89–1.74(m,4H),1.74–1.63(m,4H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-77.15,-123.17。

Step 5'

To a mixed solution of compounds 15-3b (300.00 mg, 0.435 mmol, 1.00 eq) in water/tetrahydrofuran (1/4, 5 mL) was added N-methylmorpholine oxide (76.37 mg, 0.652 mmol, 1.5 eq) and potassium osmate dihydrate (64.05 mg, 0.174 mmol, 0.4 eq) with zero degrees Centigrade stirring. The mixture was reacted at zero degrees centigrade for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonia) over 30 minutes; detector UV254 nm; compound 15-4b was obtained (pink solid, 220 mg, 69% yield). MS (ESI, m/z) 724.4/726.3[ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.82(d,J＝1.7Hz,1H),7.77(d,J＝8.3Hz,1H),7.44(t,J＝7.6Hz,1H),7.33(d,J＝7.7Hz,2H),7.28–7.21(m,1H),7.17–7.13(m,1H),5.06–4.84(m,2H),4.60–4.40(m,2H),4.21–4.04(m,1H),4.02–3.88(m,1H),3.68–3.50(m,2H),3.48–3.25(m,3H),2.80–2.54(m,2H),2.53–2.40(m,2H),2.34–2.26(m,3H),2.16–1.85(m,7H),1.76–1.63(m,3H).1.52(s,9H)。

Step 6'

To a mixed solution of compound 15-4b (70.00 mg, 0.092 mmol, 1.00 equiv) in water/tetrahydrofuran (1/4,5 ml) was added sodium periodate (117.83 mg, 0.551 mmol, 6.00 equiv) with stirring at zero degrees centigrade. The mixture was reacted at 25 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was diluted with water (50 ml) and extracted with dichloromethane (50 ml × 3). The combined organic phases were dried over anhydrous sodium sulfate, the drying agent was removed by filtration and the filtrate was concentrated to give the crude compound 15-5b (yellow solid, 69 mg). The crude product was used directly in the next reaction without further purification. MS (ESI, m/z) 692.3/694.3[ M ] +H] ⁺ 。

Step 7'

To a solution of compound 15-5b (69.00 mg, 0.095 mmol, 1.00 eq), acetic acid (5.99 mg, 0.100 mmol, 1.0 eq) and sodium cyanoborohydride (12.53 mg, 0.199 mmol, 2.0 eq) in methanol (2 ml) was added dropwise a solution of compound 1-4 (37.09 mg, 0.090 mmol, 0.90 eq) in methanol (1 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonia) over 30 minutes; detector UV254 nm; compound 15-6b was obtained (yellow solid, 45 mg, 43% yield). MS (ESI, m/z) 1089.5/1091.5[ 2 ], [ M + H ] ] ⁺ 。

Step 8'

To a solution of compound 15-6b (45.00 mg, 0.039 mmol, 1.00 eq) in dichloromethane (3 ml) was added trifluoroacetic acid (1 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; a mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: methanol; flow rate: 25 ml/min; the gradient eluted with a 25% → 54% B phase gradient over 7 minutes; detector UV254/220 nm; yield 15b (light yellow solid, 14.8 mg, 25% yield). MS (ESI, M/z): [ M + H] ⁺ ＝989.6/991.5； ¹ H NMR(300MHz,CD ₃ OD)δ8.09(d,J＝1.7Hz,1H),7.78(d,J＝8.3Hz,1H),7.61–7.55(m,1H),7.48–7.40(m,1H),7.30(d,J＝2.5Hz,1H),7.22(d,J＝5.8Hz,2H),7.12–7.03(m,3H),5.27–5.16(m,2H),5.12–5.01(m,1H),4.70–4.57(m,2H),3.74(d,J＝12.7Hz,2H),3.53–3.44(m,3H),3.44–3.37(m,3H),3.30–3.21(m,5H),3.16–2.98(m,6H),2.95(s,3H),2.91–2.67(m,6H),2.46–2.27(m,4H),2.22–2.07(m,3H),1.93–1.68(m,8H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-77.20,-123.15。

Example 16

4- ((3- (4- (3- ((S or R) -4- ((1r, 5s) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) propyl) piperazin-1-yl) propyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione tetrahydride 16.

Step 1:

to a solution of 1-bromo-3-hydroxynaphthalene (10 g, 44.38 mmol, 1.0 eq) in dichloromethane (100 ml) was added N, N-diisopropylethylamine (8.69 g, 66.57 mmol, 1.5 eq) and chloromethyl methyl ether (4.69 g, 57.69 mmol, 1.3 eq) with stirring at 25 degrees celsius under nitrogen. The mixture was reacted at 25 ℃ for 3 hours, and the progress of the reaction was monitored by liquid mass. After the reaction is finished, concentrating the reaction solution to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 10% ethyl acetate/petroleum ether and the fractions were freed from the solvent by rotary evaporation under reduced pressure to give compound 16-1 (white solid, 10.5 g, 87% yield). ¹ H NMR(300MHz,CDCl ₃ )δ8.16–8.11(m,1H),7.75–7.71(m,1H),7.57(d,J＝2.4Hz,1H),7.50–7.43(m,2H),7.39(d,J＝2.4Hz,1H),5.28(s,2H),3.52(s,3H)。

And 2, step:

to a solution of 16-1 (10 g, 35.56 mmol, 1.0 eq) in 1, 4-dioxane (100 ml) was added potassium acetate (14.70 g, 142.26 mmol, 4.0 eq), pinacol diboride (12.36 g, 46.23 mmol, 1.3 eq) and [1,1' -bis (diphenylphosphino) ferrocene ] sequentially with stirring at 25 deg.C under nitrogen protection]Palladium dichloride (3.05 g, 3.55 mmol, 0.1 eq). The mixture was reacted at 100 ℃ for 1 hour under nitrogen atmosphere and the reaction process was monitored by liquid quality. After the reaction is finished, the mixture is cooled to 25 ℃, and a crude product is obtained by decompression and concentration. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 20% ethyl acetate/petroleum ether and the fractions were freed from the solvent by rotary evaporation under reduced pressure to give compound 16-2 (white solid, 10 g, 85% yield). MS (ESI, m/z) 315.2[ m ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ8.72–8.66(m,1H),7.82(d,J＝2.7Hz,1H),7.79–7.73(m,1H),7.51(d,J＝2.7Hz,1H),7.49–7.40(m,2H),5.33(s,2H),3.54(s,3H),1.44(s,12H)。

And step 3:

to a 250 ml three-necked flask, under nitrogen protection at 25 degrees celsius, were added compound 3-2 (9 g, 16.89 mmol, 1.0 eq), 16-2 (5.59 g, 16.89 mmol, 1.0 eq), 1, 4-dioxane (80 ml), water (20 ml), sodium carbonate (3.77 g, 33.78 mmol, 2.0 eq), tris-dibenzylideneacetone dipalladium (0) (775 mg, 0.85 mmol, 0.05 eq), and 4- (2, 6-dimethoxyphenyl) -3-tert-butyl-2, 3-dihydro-1, 3-benzoxap-hosterocycle (558 mg, 1.69 mmol, 0.1 eq), in that order. The mixture was reacted at 60 degrees celsius for 2 hours under nitrogen atmosphere. The reaction process is monitored by liquid quality. After the reaction is finished, concentrating the reaction solution to obtain a crude product. The crude product obtained is purified by chromatography on a silica gel column, the mobile phase is eluted with a gradient of 0% → 20% ethyl acetate/petroleum ether and the fractions obtained are freed from the solvent by rotary evaporation under reduced pressure to give the crude product. The crude product is then purified by means of a reverse phase chromatography column (C18 column) eluting within 20 minutes with 40% → 90% acetonitrile/water mobile phase (0.1% ammonium bicarbonate); detector UV254/220 nm; compound 16-3 was obtained (racemic mixture of two stereoisomers, yellow solid, 6 g, 55% yield). MS (ESI, m/z) 613.2/615.2[ M + H ] ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.88–7.85(m,2H),7.58(d,J＝2.5Hz,1H),7.53–7.47(m,1H),7.32–7.30(m,2H),7.21(d,J＝2.4Hz,1H),5.36(s,2H),4.56–4.41(m,4H),3.77–3.65(m,2H),3.58(s,3H),2.06–2.00(m,2H),1.87–1.79(m,2H),1.56(s,9H)。

And 4, step 4:

the compound 16-3 (6 g) from step 3 was subjected to preparative supercritical liquid chromatographySexual resolution: chiral column NB — CHIRALPAK IC,5 × 25 cm, 5 microns; a mobile phase A: supercritical carbon dioxide fluid, mobile phase B: ethanol; flow rate: 160 ml/min; column temperature: 35 ℃; elution with 45% mobile phase B for 15 min; detector UV225 nm. Two products were obtained, with a shorter retention time (9.03 minutes) of compound 16-3a (yellow solid, 2.5 g, 41% recovery), compound 16-3a MS (ESI, m/z): 613.2/615.2[ M ] +H ]] ⁺ (ii) a The product of longer retention time (10.78 minutes) was compound 16-3b (yellow solid, 2.9 g, recovery 48%), compound 16-3b] ⁺ 。

Step 5

To a solution of compound 16-3a (300.0 mg, 0.47 mmol, 1.0 equiv) and 3-butene-1-ol (42.3 ml, 0.56 mmol, 1.2 equiv) in tetrahydrofuran (5.0 ml) was added dropwise a 1 mol/l solution of potassium tert-butoxide in tetrahydrofuran (0.59 ml, 0.56 mmol, 1.2 equiv) at zero degrees centigrade, with stirring under nitrogen. The resulting mixture was stirred at zero degrees centigrade for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 16-4a (off-white solid, 230.0 mg, 72% yield). MS (ESI, m/z) 649.3/651.3[ m + H ], [ solution ] M + ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.89–7.79(m,2H),7.56(d,J＝2.4Hz,1H),7.52–7.45(m,1H),7.39–7.29(m,2H),7.22(d,J＝2.5Hz,1H),6.02–5.88(m,1H),5.40–5.33(m,2H),5.25–5.08(m,2H),4.56–4.36(m,6H),3.59–3.57(m,5H),2.71–2.57(m,2H),2.07–2.01(m,2H),1.91–1.87(m,2H),1.55(s,9H)。

Step 6

To a solution of compound 16-4a (225.0 mg, 0.33 mmol, 1.0 equiv) in acetone/water (4/1, 5.0 mL) was added N-methylmorpholine oxide (60.9 mg, 0.49 mmol, 1.5 equiv) and potassium osmate dihydrate (12.7 mg, 0.33 mmol, 0.1 equiv) with zero degrees Centigrade stirring. The resulting mixture was stirred at 25 ℃ for an additional 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was directly purified by a reverse phase flash column (C18 column), and eluted with 40% → 95% methanol/water mobile phase (0.1% ammonia) within 20 minutes; detector UV254 nm; compound 16-5a was obtained (off-white solid, 180 mg, 76% yield). MS (ESI, m/z) 683.3/685.3[ M ] +H] ⁺ 。

Step 7

To a solution of compound 16-5a (60.0 mg, 0.08 mmol, 1.0 eq) in acetonitrile/water (4/1,2.5 ml) was added sodium periodate (107.1 mg, 0.5 mmol, 6.0 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, water (10.0 ml) was added to the reaction solution, the mixture was extracted with dichloromethane (10.0 ml × 3), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give a crude product 16-6a (pale yellow oily liquid, 62.0 mg) which was used directly in the next reaction. MS (ESI, m/z) 651.3/653.3[ 2 ], [ M + H ] ] ⁺ 。

Step 8

To compound 9-6 (53.8 mg, 0.11 mmol, 1.2 eq), sodium acetate (15.6 mg, 0.19 mmol, 2.0) was added with stirring at 25 deg.cEq), acetic acid (17.2 mg, 0.29 mmol, 3.0 eq) and sodium cyanoborohydride (18.0 mg, 0.29 mmol, 3.0 eq) in methanol (2.0 ml) was added dropwise a solution of compound 16-6a (62.0 mg, 0.10 mmol, 1.0 eq) in methanol (0.5 ml). The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 16-7a (yellow solid, 37 mg, yield 35%) was obtained. MS (ESI, m/z) 1105.6/1107.6[ m + H ]] ⁺ 。

Step 9

To a solution of compound 16-7a (37.0 mg, 0.03 mmol, 1.0 eq) in methanol (2.0 ml) was added a solution of 4 mol/l hydrochloric acid in 1, 4-dioxane (2.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X150 mm, 5 μm; a mobile phase A: water (0.05% hydrochloric acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 30% b phase gradient over 7 minutes; detector UV 254/220 nm; compound 16 was obtained (yellow solid, 24.0 mg, 64% yield). MS (ESI, m/z) 961.4/963.4[ m ] +H ] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ Heavy water exchange) δ 8.05 (s, 1H), 7.89-7.86 (d, J =8.3hz, 1H), 7.70-7.66 (m, 1H), 7.55-7.50 (m, 1H), 7.38-7.37 (m, 1H), 7.33-7.21 (m, 3H), 7.14-7.12 (m, 2H), 5.13-5.07 (m, 1H), 4.65-4.47 (m, 4H), 4.25 (s, 2H), 3.96-3.88 (m, 4H), 3.52-3.48 (m, 4H), 3.39-3.24 (m, 7H), 3.18-3.08 (m, 1H), 2.99-2.77 (m, 6H), 2.75-2.70 (m, 1H), 2.39-2.32 (m, 2.31H), 2.93-2.93 (m, 1H); ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-122.00。

example 17

4- ((15- (((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -12-methyl-3, 6, 9-trioxa-12-azapentadecyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione formate 17a;4- ((15- (((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -12-methyl-3, 6, 9-trioxa-12-azapentadecyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindole-1, 3-dione formate 17b.

Step 1

The compound 2- (2, 6-dioxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (2.00 g, 7.241 mmol, 1.00 eq), 1-amino-3, 6, 9-trioxa-11-undecanol (1.54 g, 7.969 mmol, 1.10 eq), N-dimethylformamide (20 ml) and N, N-diisopropylethylamine (1.87 g, 14.481 mmol, 2.00 eq) were added to the reaction flask in that order with stirring at zero degrees celsius. The resulting mixture was stirred at 90 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature. The mixture was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 8% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 17-1 (yellow oil, 701 mg, yield 21%) was obtained. MS (ESI, m/z) 450.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.16(s,1H),7.52–7.47(m,1H),7.11(d,J＝7.1Hz,1H),6.93(d,J＝8.5Hz,1H),4.95–4.89(m,1H),3.76–3.66(m,12H),3.64–3.59(m,2H),3.48(t,J＝5.3Hz,2H),2.92–2.85(m,1H),2.82–2.71(m,2H),2.18–2.10(m,1H)。

Step 2

To a solution of compound 17-1 (170.00 mg, 0.367 mmol, 1.00 eq) in dichloromethane (150.0 ml) was added 1, 1-triacetoxy-1, 1-dihydro-1, 2-phenyliodoxy-3 (1H) -one (327.60 mg, mmol, 2.00 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the insoluble material was removed by filtration and the filter cake was washed with dichloromethane (3 × 20 ml). And concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 17-2 (pale yellow oil, 140 mg, 70% yield). MS (ESI, m/z) 448.3[ 2 ], [ M ] +H] ⁺ 。

Step 3

To a 25 ml single neck flask were added, in order, compound 2-9a (25 mg, 0.042 mmol, 1.0 eq), 17-2 (56.3 mg, 0.126 mmol, 3.0 eq), methanol (2.0 ml), acetic acid (3.78 mg, 0.063 mmol, 1.5 eq) and sodium cyanoborohydride (3.95 mg, 0.063 mmol, 1.5 eq) at 25 ℃. The resulting mixture was reacted at 40 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the excessive solvent is removed by decompression and concentration to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% trifluoroacetic acid) over 20 minutes; detector UV254 nm; compound 17-3a was obtained (yellow oily liquid, 23 mg, 48% yield). MS (ESI, m/z) 1027.4/1029.4[ 2 ], [ M ] +H ] ⁺ 。

Step 4

Compound 17-3a (20 mg, 0.018 mmol, 1.0 eq), dichloromethane (2.0 ml) and trifluoroacetic acid (1.0 ml) were added sequentially to a 25 ml single-neck flask at 0 ℃. The resulting mixture was reacted at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the excessive solvent is removed by decompression and concentration to obtain a crude product. Purifying the crude product by a preparative high performance liquid chromatograph under the following purification conditions: xsselect CSH Prep C18 OBD Column,19 × 250 mm, 5 μm, mobile phase a: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 31% b phase gradient over 7 minutes; detector UV 254/220 nm; compound 17a was obtained (yellow solid, 10 mg, 56% yield). MS (ESI, m/z) 927.4/929.4; ¹ H NMR(300MHz,DMSO-d ₆ )δ11.11(s,1H),10.30–9.80(m,1H),8.18(s,1H),7.94(d,J＝1.6Hz,1H),7.81(d,J＝8.3Hz,1H),7.59–7.52(m,1H),7.48–7.40(m,1H),7.28(d,J＝2.4Hz,1H),7.25–7.17(m,2H),7.13–6.99(m,3H),6.62–6.53(m,1H),5.11–4.99(m,1H),4.35(t,J＝6.6Hz,2H),3.85–3.77(m,4H),3.59–3.56(m,2H),3.55–3.46(m,9H),3.44–3.25(m,6H),3.03–2.96(m,4H),2.93–2.78(m,2H),2.63–2.53(m,2H),2.19(s,3H),2.08–1.96(m,1H),1.93–1.79(m,2H)。

step 3'

Compounds 2-8b (760 mg, 1.1 mmol, 1.0 eq), 1, 3-dimethyl barbituric acid (279.8 mg, 1.7 mmol, 1.5 eq), tetrakis (triphenylphosphine) palladium (69.0 mg, 0.057 mmol, 0.05 eq) and dichloromethane (10 ml) were added sequentially to a 50 ml three-neck flask under nitrogen protection at 25 ℃, and the reaction was monitored by liquid chromatography and thin layer chromatography at 25 ℃ for 5 hours. After the reaction is finished, concentrating under reduced pressure to remove the redundant solvent to obtain a crude product. Crude product The product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% formic acid) over 20 minutes, detector UV254 nm to give compounds 2-9b (light yellow solid, 640 mg, 89% yield). MS (ESI, m/z) 596.2/598.2[ M + H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.61(s,1H),7.69–7.66(m,2H),7.35–7.31(m,1H),7.28–7.27(m,1H),7.18–7.08(m,3H),4.52–4.49(m,2H),3.82–3.64(m,8H),3.26(brs,2H),3.00–2.91(m,2H),2.50(s,3H),2.21–2.18(m,2H),1.49(s,9H)。

Step 4'

To a 25 ml single neck flask were added, in order, compound 2-9b (25 mg, 0.04 mmol, 1.0 eq), 17-2 (51.8 mg, 0.12 mmol, 3.0 eq), methanol (2.0 ml), acetic acid (3.78 mg, 0.06 mmol, 1.5 eq), and sodium cyanoborohydride (3.95 mg, 0.063 mmol, 1.5 eq) at 25 ℃. The resulting mixture was reacted at 40 degrees celsius for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the excessive solvent is removed by decompression and concentration to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% trifluoroacetic acid) over 20 minutes; detector UV254 nm; compound 17-3b was obtained (yellow oily liquid, 25 mg, 54% yield). MS (ESI, m/z) 1027.4/1029.4[ 2 ], [ M ] +H] ⁺ 。

Step 5'

Compound 17-3b (20 mg, 0.018 mmol, 1.0 eq), dichloromethane (2.0 ml) and trifluoroacetic acid (1.0 ml) were added sequentially to a 25 ml single-neck flask at 0 degrees celsius under air. The resulting mixture was reacted at 25 ℃ for 2 hours. The reaction process is monitored by liquid quality. After the reaction is finished, decreasing Concentrating under reduced pressure to remove excess solvent to obtain crude product. Purifying the crude product by a preparative high performance liquid chromatograph under the following conditions: xsselect CSH Prep C18 OBD Column,19 × 250 mm, 5 μm, mobile phase a: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 31% b phase gradient over 7 minutes; detector UV 254/220 nm; compound 17b was obtained (yellow solid, 10 mg, 57% yield). MS (ESI, m/z) 927.4/929.4; ¹ H NMR(300MHz,DMSO-d ₆ )δ11.19(s,1H),10.30–9.80(m,1H),8.26(s,1H),8.02(d,J＝1.6Hz,1H),7.88(d,J＝8.3Hz,1H),7.67–7.58(m,1H),7.56–7.47(m,1H),7.36(d,J＝2.4Hz,1H),7.34–7.24(m,2H),7.20–7.06(m,3H),6.71–6.58(m,1H),5.18–5.06(m,1H),4.50–4.37(m,2H),3.92–3.84(m,4H),3.68–3.63(m,3H),3.61–3.54(m,5H),3.52–3.28(m,9H),3.16–3.04(m,4H),2.98–2.84(m,2H),2.69–2.60(m,2H),2.27(s,3H),2.15–2.02(m,1H),2.00–1.88(m,2H)。

example 18

(2s, 4r) -1- ((S) -2- (tert-butyl) -18- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -15-methyl-4-oxo-6, 9, 12-trioxy-3, 15-diazadecyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 18a; (2S, 4R) -1- ((S) -2- (tert-butyl) -18- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -15-methyl-4-oxo-6, 9, 12-trioxy-3, 15-diazacyclo-decyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 18b.

Step 1

To the compound triethylene glycol (20 g, 126.52 mm) was added under stirring at zero degrees centigrade and nitrogen blanket Mol, 1.00 eq) in dichloromethane (400 ml) was added dropwise a solution of boron trifluoride in diethyl ether (0.40 ml, 2.824 mmol, 0.05 eq) and ethyl diazoacetate (7.6 g, 63.26 mmol, 0.50 eq). The mixture was allowed to react at 25 ℃ for 2 hours with stirring under nitrogen, the course of the reaction being monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with 33% dichloromethane/ethyl acetate mobile phase and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 18-1 (pale yellow oil, 4 g, 14% yield). MS (ESI, m/z) 237.2[ deg. ] M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.14(q,J＝7.1Hz,2H),4.07(s,2H),3.71–3.61(m,6H),3.60(s,4H),3.55–3.49(m,2H),2.88(s,1H),1.21(t,J＝7.1Hz,3H)。

Step 2

To a solution of compound 18-1 (4.00 g, 16.94 mmol, 1.00 eq) and N, N-diisopropylethylamine (3.00 g, 22.02 mmol, 1.3 eq) in dichloromethane (20 ml) was added dropwise methanesulfonyl chloride (2.44 g, 20.33 mmol, 1.2 eq) at zero degrees centigrade, with stirring under nitrogen protection. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 18-2 (brown oil, 3.6 g, 67% yield). MS (ESI, m/z) 315.1[ m ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.37–4.32(m,2H),4.17(q,J＝7.1Hz,2H),4.09(s,2H),3.76–3.71(m,2H),3.71–3.57(m,8H),3.04(s,3H),1.25(t,J＝7.1Hz,3H)。

Step 3

To a solution of compound 18-2 (1.80 g, 5.726 mmol, 1.00 eq) and 3- (methylamino) -1-propanol (641.19 mg, 6.834 mmol, 1.30 eq) in acetonitrile (18 ml) was added potassium carbonate (1.53 g, 10.513 mmol, 2.00 eq) at 25 ℃ with stirring under nitrogen. The mixture was reacted at 90 ℃ for 16 hours with stirring under nitrogen protection, and the course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the reaction was cooled to 25 ℃. The reaction solution was directly purified by reverse phase flash chromatography (C18 column) and eluted with 5% → 45% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 25 minutes; detector UV200 nm; compound 18-3 was obtained (colorless oil, 800 mg, 39% yield). MS (ESI, m/z) 308.1[ 2 ], M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.23(q,J＝7.1Hz，2H),4.16(s,2H),3.83–3.58(m,12H),2.69–2.59(m,4H),2.31(s,3H),1.77–1.64(m,2H),1.30(t,J＝7.1Hz,3H)。

Step 4

Cesium fluoride (395.38 mg, 2.47 mmol, 1.50 eq), 18-3 (582.43 mg, 1.65 mmol, 1.0 eq), and N, N-diisopropylethylamine (336.40 mg, 2.47 mmol, 1.50 eq) were added to a solution of compounds 1-7 (800 mg, 1.65 mmol, 1.00 eq) in N-methylpyrrolidinone (10 ml) at 25 ℃ with stirring under nitrogen. The resulting mixture was stirred at 100 ℃ for 3 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature. The reaction solution was directly purified by reverse phase flash chromatography (C18 column) eluting with 20% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 20 minutes; detector UV254 nm; compound 18-4 was obtained (brown oil, 250 mg, 19% yield). MS (ESI, m/z) 750.2/752.1/754.1M + H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.72(d,J＝2.0Hz,1H),4.52(t,J＝6.5Hz,2H),4.23(q,J＝7.2Hz,2H),4.16(s,2H),3.83–3.59(m,18H),2.86–2.57(m,4H),2.51–2.27(m,3H),2.03(s,2H),1.52(s,9H),1.30(t,J＝7.1Hz,3H)。

Step 5

To a mixed solution of compound 18-4 (250 mg, 0.28 mmol, 1.00 eq) in water/methanol/tetrahydrofuran (1/1, 1.5 ml) was added lithium hydroxide (34.65 mg, 1.38 mmol, 5.0 eq) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was purified by reverse phase flash chromatography (C18 column) and eluted with acetonitrile/water mobile phase (0.1% ammonium bicarbonate) of 5% → 95% in 20 minutes; detector, UV254 nm; compound 18-5 was obtained (colorless oil, 210 mg, 89% yield). MS (ESI, m/z) 722.1/724.2/726.1M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.69(d,J＝2.0Hz,1H),4.57–4.49(m,2H),4.01(s,2H),3.86–3.73(m,6H),3.73–3.57(m,12H),3.11–2.96(m,4H),2.64(s,3H),2.29–2.15(m,2H),1.49(s,9H)。

Step 6

To a solution of compound 18-5 (210 mg, 0.27 mmol, 1.00 eq) and 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (131.41 mg, 0.33 mmol, 1.2 eq) in N, N-dimethylformamide (2.0 ml) was added (2s, 4r) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) with stirring at 25 degrees celsius]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] amide]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (138.54 mg, 0.27 mmol, 1.0 eq) and N, N-diisopropylethylamine (148.89 mg, 1.09 mmol, 4.0 eq). The mixture was reacted at 25 ℃ for 2 hours, the course of the reaction being monitored by liquid chromatography and thin-layer chromatography . After completion of the reaction, the reaction solution was purified by reverse phase flash chromatography (C18 column) and eluted with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 11 minutes; detector UV254 nm; compound 18-6 was obtained (off-white solid, 260 mg, 77% yield). MS (ESI, m/z) 1148.4/1150.3/1152.3[ m ] +H] ⁺ 。

Step 7

To a solution of compound 18-6 (230 mg, 0.18 mmol, 1.00 eq) in 1, 4-dioxane/water (1/1, 6 ml) was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (61.44 mg, 0.216 mmol, 1.2 eq), sodium carbonate (57.25 mg, 0.51 mmol, 2.85 eq) and [1,1' -bis (diphenylphosphino) ferrocene ] at 25 deg.C with stirring under nitrogen protection]Palladium dichloride dichloromethane complex (II) (15.44 mg, 0.02 mmol, 0.10 eq). The reaction solution was stirred at 90 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the reaction solution was cooled to 25 ℃. The reaction solution was concentrated and purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 30 minutes; detector UV254 nm; compound 18-7 was obtained (off-white solid, 140 mg, 57% yield). MS (ESI, m/z) 1212.4/1214.4[ M ] +H ] ⁺ 。

Step 8

Chiral resolution of the compound 18-7 (140 mg) obtained in step 7 by preparative chiral high performance liquid chromatography: chiral column CHIRALPAK IC,2 × 25 cm, 5 μm; a mobile phase A: n-hexane/dichloromethane =3/1 (0.1% diethylamine), mobile phase B: ethanol; flow rate: 15 ml/min; elution was performed with 50% of phase B in 26 minutes, detector UV220/235 nm. Two products were obtained, with a shorter retention time (8.11 minutes) being compound 18-7a (off-white solid, 52 mg, 37% recovery), compound 18-7a: MS (ESI, m/z) 1212.4/1214.4[ M ] +H] ⁺ (ii) a The product of longer retention time (18 min) was compound 18-7b (off-white solid, 57 mg, 40% recovery), compound 18-7b: MS (ESI, m/z) 1212.4/1214.4[ M ] +H] ⁺ 。

Step 9

To a solution of compound 18-7a (52.00 mg, 0.043 mmol, 1.00 eq) in dichloromethane (1.5 ml) was added trifluoroacetic acid (0.5 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 0.5 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: gemini-NX C18 AXAI Packed,21.2x150 mm, 5 microns; a mobile phase A: water (10 mmol/l ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: eluting with 10% B phase within 2 minutes, followed by 10% → 36% B phase gradient within 2.5 minutes, and finally 36% → 65% B phase gradient within 10.5 minutes; detector UV 220 nm; yield 18a (off-white solid, 24.3 mg, 49% yield). MS (ESI, M/z) 1112.6/1114.6.[ M + H ] ] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ ):δ10.02(s,1H),8.99(s,1H),8.44(d,J＝7.6Hz,1H),7.92(d,J＝1.6Hz,1H),7.81(d,J＝8.3Hz,1H),7.50–7.32(m,6H),7.29(d,J＝2.4Hz,1H),7.22(d,J＝4.0Hz,2H),7.07(d,J＝2.4Hz,1H),5.14(s,1H),4.90(t,J＝7.1Hz,1H),4.54(d,J＝9.5Hz,1H),4.44(t,J＝8.2Hz,1H),4.36(t,J＝6.5Hz,2H),4.28(s,1H),3.94(s,2H),3.78(d,J＝5.7Hz,4H),3.63–3.41(m,13H),2.90(d,J＝5.0Hz,4H),2.53–2.35(m,7H),2.45(s,3H),2.19(s,1H),2.12–1.97(m,3H),1.37(d,J＝7.0Hz,3H),0.93(s,9H)。

Step 10

Trifluoroacetic acid (0.5 ml) was added to a solution of compounds 18-7B ((57.00 mg, 0.047 mmol, 1.00 eq) in dichloromethane (1.5 ml) with stirring at 25 degrees celsius, the reaction was monitored at 25 degrees celsius for 0.5 hour, and the progress of the reaction was monitored by liquid and thin layer chromatography after completion of the reaction the crude product was purified by preparative high performance liquid chromatography under conditions Gemini-NX C18 AXAI Packed,21.2x150 mm, 5 μm, mobile phase a: water (10 mmol/l ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with 10% B phase in 2 minutes, followed by 10% B phase gradient in 2.5 minutes, elution with 36% B phase gradient, and finally elution with 36% B phase gradient in 10.5 minutes → 65%, UV detector 220 nm,: 18B (type white solid, 24.1112.45, 6.1114/z); ¹ H NMR(300MHz,DMSO-d ₆ ):δ10.02(s,1H),8.99(s,1H),8.44(d,J＝7.6Hz,1H),7.92(d,J＝1.6Hz,1H),7.81(d,J＝8.3Hz,1H),7.50–7.32(m,6H),7.29(d,J＝2.4Hz,1H),7.22(d,J＝4.0Hz,2H),7.07(d,J＝2.4Hz,1H),5.14(s,1H),4.90(t,J＝7.1Hz,1H),4.54(d,J＝9.5Hz,1H),4.44(t,J＝8.2Hz,1H),4.36(t,J＝6.5Hz,2H),4.28(s,1H),3.94(s,2H),3.78(d,J＝5.7Hz,4H),3.63–3.41(m,13H),2.90(d,J＝5.0Hz,4H),2.53–2.35(m,7H),2.45(s,3H),2.19(s,1H),2.12–1.97(m,3H),1.37(d,J＝7.0Hz,3H),0.93(s,9H)。

example 19

(2s, 4r) -1- ((S) -2- (tert-butyl) -21- ((6-chloro-8-fluoro-7- ((S or R) -3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -18-methyl-4-oxo-6, 9,12, 15-tetraoxo-3, 18-diazaspiro-lyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 19a; (2S, 4R) -1- ((S) -2- (tert-butyl) -21- ((6-chloro-8-fluoro-7- ((R or S) -3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -18-methyl-4-oxo-6, 9,12, 15-tetraoxo-3, 18-diazaspiro-lyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 19b.

Step 1

To a solution of the compound tetraethylene glycol (20 g, 97.82 mmol, 2 equiv) in dichloromethane (300 ml) at zero degrees centigrade under nitrogen protection with stirring was added dropwise an ether solution of boron trifluoride (9.46 mg, 0.06 mmol, 0.01 equiv) and ethyl diazoacetate (5.88 g, 48.96 mmol, 1.00 equiv). The mixture was allowed to react at 25 ℃ for 0.5 h with stirring under nitrogen, the course of the reaction being monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with 0% → 10% methanol/dichloromethane mobile phase and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 19-1 (yellow oil, 4.92 g, yield 35%). MS (ESI, m/z) 281.2[ m ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.19(q,J＝7.1Hz,2H),4.12(s,2H),3.75–3.61(m,14H),3.61–3.54(m,2H),1.26(t,J＝7.1Hz,3H)。

Step 2

To a solution of compound 19-1 (4.00 g, 13.556 mmol, 1.00 eq) and N, N-diisopropylethylamine (2.95 g, 21.690 mmol, 1.60 eq) in dichloromethane (30 ml) was added dropwise methanesulfonyl chloride (2.12 g, 17.623 mmol, 1.30 eq) at zero degrees centigrade, with stirring under nitrogen. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography with 0% → 10% methanol/dichloromethane The alkyl mobile phase was subjected to gradient elution, and the obtained fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby Compound 19-2 (pale yellow oil, 4.15 g, yield 81%) was obtained. MS (ESI, m/z) 359.1[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.41–4.34(m,2H),4.21(q,J＝7.2Hz,2H),4.14(s,2H),3.80–3.60(m,14H),3.08(s,3H),1.28(t,J＝7.1Hz,3H)。

Step 3

To a solution of compound 19-2 (4.15 g, 11.000 mmol, 1.00 eq) and 3- (methylamino) -1-propanol (1.24 g, 13.200 mmol, 1.20 eq) in acetonitrile (50 ml) was added potassium carbonate (3.20 g, 22.001 mmol, 2.00 eq) with stirring at 25 ℃. The mixture was reacted at 80 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature. The insolubles were removed by filtration and the filter cake was washed with dichloromethane (50 ml x 3). And concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 13% methanol (3% ammonia)/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, giving a crude product. The crude product was further purified by reverse phase flash chromatography (C18 column) eluting with 5% → 50% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 15 minutes; detector, UV205 nm; compound 19-3 was obtained (pale yellow oil, 1.3 g, 31% yield). MS (ESI, m/z) 352.2[ 2 ], [ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.21(q,J＝7.2Hz,2H),4.14(s,2H),3.82–3.75(m,2H),3.74–3.67(m,4H),3.67–3.57(m,10H),2.68–2.60(m,4H),2.32(s,3H),1.74–1.66(m,2H),1.28(t,J＝7.1Hz,3H)。

Step 4

Adding the mixture into a compound 1 at 25 ℃ under the stirring protection of nitrogen-7 (910.00 mg, 1.895 mmol, 1.00 eq) of N-methylpyrrolidinone (8 ml) cesium fluoride (431.83 mg, 2.843 mmol, 1.50 eq), 19-3 (582.43 mg, 1.65 mmol, 1.0 eq) and N, N-diisopropylethylamine (489.88 mg, 3.790 mmol, 2.00 eq) are added. The resulting mixture was stirred at 120 ℃ for 3 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature. The reaction solution was directly purified by reverse phase flash chromatography (C18 column) and eluted with 5% → 100% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 20 minutes; detector UV254 nm; compound 19-4 was obtained (brown oil, 350 mg, 22% yield). MS (ESI, m/z) 794.3/796.3/798.3[ 2 ] M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.70(d,J＝2.0Hz,1H),4.50(t,J＝6.5Hz,2H),4.21(q,J＝7.1Hz,2H),4.14(s,2H),3.81–3.59(m,22H),2.78–2.52(m,4H),2.42–2.26(m,3H),2.05(d,J＝6.3Hz,2H),1.50(s,9H),1.28(t,J＝7.2Hz,3H)。

Step 5

To a solution of compound 19-4 (310.00 mg, 0.370 mmol, 1.00 eq) in water/methanol/tetrahydrofuran (1/1, 3 ml) was added lithium hydroxide (18.67 mg, 0.741 mmol, 2 eq) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, diluted hydrochloric acid (1 mol/l, 88. Mu.l) was added to the reaction mixture to neutralize it. The resulting mixture was concentrated under reduced pressure to give a crude acid. The crude acid was dissolved in N, N-dimethylformamide (2.0 ml) and 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (177.89 mg, 0.444 mmol, 1.2 eq) was added. The mixture was reacted at 25 ℃ for 1 hour. (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) was added to the above-mentioned reaction mixture ]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (190.66 mg, 0.407 mmol)Mol, 1.1 eq) and N, N-diisopropylethylamine (201.55 mg, 1.482 mmol, 4.00 eq). The mixture was allowed to react at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was purified by reverse phase flash chromatography (C18 column) and eluted with 30% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 20 minutes; detector UV 254 nm; compound 19-5 was obtained (white solid, 300 mg, 67% yield). MS (ESI, m/z) 1192.3/1194.3/1196.3[ 2 ] M + H] ⁺ 。

Step 6

To a mixed solution of compound 19-5 (300.00 mg, 0.251 mmol, 1.00 eq.) in 1, 4-dioxane/water (5/1, 3.6 mL) was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (101.84 mg, 0.377 mmol, 1.50 eq.), sodium carbonate (53.28 mg, 0.503 mmol, 2.00 eq.) and [1,1' -bis (diphenylphosphino) ferrocene ] at 25 deg.C with stirring under nitrogen protection]Palladium dichloride dichloromethane complex (II) (20.47 mg, 0.025 mmol, 0.10 eq). The reaction solution was stirred at 80 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, cooling the reaction liquid to 25 ℃, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a methanol/dichloromethane mobile phase gradient of 0% → 10% and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 19-6 (white solid, 280 mg, 88% yield). MS (ESI, m/z) 1256.5/1258.6[ m ] +H ] ⁺ 。

Step 7

Chiral resolution of the compound 19-6 (280 mg) obtained in step 6 by preparative chiral high performance liquid chromatography: a chiral column CHIRALPAK IC,2x25 cm, 5 microns; mobile phase A: n-hexane/dichloromethane =3/1 (0.1% diethylamine), mobile phase B: ethanol; flow rate: 15 ml/min; elution was performed with 50% phase B over 32 minutes, detector UV220/254 nm. Two products were obtained, with a shorter retention time (12.06 minutes) being compound 19-6a (off-white solid, 130 mg, 48% recovery), compound 19-6a: MS (ESI, m/z) 1256.5/1258.6[ 2 ], [ M ] +H] ⁺ (ii) a The product of longer retention time (23.776 min) was compound 19-6b (off-white solid, 120 mg, 45% recovery), compound 19-6b: MS (ESI, m/z) 1256.5/1258.6[ m ] +H] ⁺ 。

Step 8

To a solution of compound 19-6a (60.00 mg, 0.048 mmol, 1.00 eq) in dichloromethane (1 ml) was added trifluoroacetic acid (0.3 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: gemini-NX C18 AXAI Packed,21.2x150 mm, 5 microns; a mobile phase A: water (10 mmol/l ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: eluting with 26% → 55% b phase gradient in 10 minutes, detector UV254/220 nm; yield 19a (white solid, 23.8 mg, 43% yield). MS (ESI, m/z) 1156.5/1158.5[ m ] +H ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.04(s,1H),8.98(s,1H),8.43(d,J＝7.6Hz,1H),7.91(d,J＝1.6Hz,1H),7.80(d,J＝8.4Hz,1H),7.47–7.40(m,3H),7.41–7.32(m,3H),7.28(d,J＝2.4Hz,1H),7.25–7.19(m,2H),7.06(d,J＝2.4Hz,1H),5.13(s,1H),4.94–4.87(m,1H),4.54(d,J＝9.6Hz,1H),4.44(t,J＝8.4Hz,1H),4.35(t,J＝6.4Hz,2H),4.30–4.25(m,1H),3.94(s,2H),3.79–3.74(m,4H),3.63–3.52(m,6H),3.51–3.49(m,4H),3.48–3.44(m,6H),2.91–2.85(m,4H),2.48–2.46(m,4H),2.45(s,3H),2.18(s,3H),2.09–2.00(m,1H),1.92–1.82(m,2H),1.81–1.73(m,1H),1.37(d,J＝7.0Hz,3H),0.93(s,9H)。

Step 8'

To a solution of compound 19-6b (60.00 mg, 0.048 mmol, 1.00 eq) in dichloromethane (1 ml) was added trifluoroacetic acid (0.3 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: gemini-NX C18 AXAI Packed,21.2x150 mm, 5 microns; a mobile phase A: water (10 mmol/l ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elute with a 31% → 65% B phase gradient over 10 minutes, detector: UV 254/220 nm; yield 19b (white solid, 23.1 mg, 41% yield). MS (ESI, m/z) 1156.5/1158.5[ m ] +H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.02(s,1H),8.98(s,1H),8.43(d,J＝7.6Hz,1H),7.91(d,J＝1.6Hz,1H),7.80(d,J＝8.4Hz,1H),7.47–7.40(m,3H),7.40–7.32(m,3H),7.28(d,J＝2.4Hz,1H),7.25–7.18(m,2H),7.06(d,J＝2.4Hz,1H),5.20–5.09(m,1H),4.93–4.87(m,1H),4.54(d,J＝9.6Hz,1H),4.48–4.40(m,1H),4.38–4.32(m,2H),4.30–4.26(m,1H),3.94(s,2H),3.84–3.70(m,4H),3.64–3.52(m,6H),3.52–3.49(m,4H),3.48–3.44(m,6H),2.92–2.84(m,4H),2.48–2.46(m,4H),2.45(s,3H),2.18(s,3H),2.06–2.02(m,1H),1.89–1.84(m,2H),1.81–1.73(m,1H),1.37(d,J＝7.2Hz,3H),0.93(s,9H)。

Example 20

(2s, 4r) -1- ((S) -2- (tert-butyl) -24- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -21-methyl-4-oxo-6, 9,12,15, 18-pentaoxo-3, 21-diazo-4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 20a; (2S, 4R) -1- ((S) -2- (tert-butyl) -24- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -21-methyl-4-oxo-6, 9,12,15, 18-pentaoxo-3, 21-diazocinoyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 20b.

Step 1

To a 250 ml single neck flask were added pentaethylene glycol (10.0 g, 41.5 mmol, 1.0 eq), boron trifluoride in ether (443.5 mg, 3.1 mmol, 0.1 eq) and dichloromethane (150.0 ml) with zero degrees centigrade stirring, followed by slow dropwise addition of ethyl diazoacetate (2.4 g, 20.7 mmol, 0.5 eq). The resulting mixture was stirred at 0 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the mixture is recovered to 25 ℃, and the mixture is concentrated under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 4% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 20-1 (yellow oil, 2.4 g, 18% yield). MS (ESI, m/z) 325.3[ 2 ], [ M ] +H] ⁺ ； ¹ HNMR(300MHz,CDCl ₃ )δ4.22(q,J＝7.1Hz,2H),4.15(s,2H),3.78–3.69(m,6H),3.68–3.65(m,12H),3.63–3.59(m,2H),1.29(t,J＝7.1Hz,3H)。

Step 2

To a solution of 20-1 (2.4 g, 7.4 mmol, 1.0 eq) in dichloromethane (50.0 ml) was added methanesulfonyl chloride (1.1 g, 9.3 mmol, 1.3 eq) and N, N-diisopropylethylamine (1.6 g, 11.4 mmol, 1.6 eq) with stirring at zero degrees centigrade.The reaction solution was reacted at 25 ℃ for 2 hours, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient, and the resulting fraction was rotary distilled under reduced pressure to remove the solvent, affording compound 20-2 (yellow oil, 2.0 g, 66% yield). MS (ESI, m/z) 403.1[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.44–4.34(m,2H),4.21(q,J＝7.1Hz,2H),4.15(s,2H),3.82–3.59(m,18H),3.09(s,3H),1.29(t,J＝7.1Hz,3H)。

Step 3

To a 100 ml single neck flask were added sequentially compound 20-2 (2.0 g, 4.7 mmol, 1.0 eq), 3- (methylamino) -1-propanol (0.5 g, 5.7 mmol, 1.2 eq), acetonitrile (50.0 ml) and potassium carbonate (1.4 g, 9.4 mmol, 2.0 eq) with stirring at 25 ℃. The mixture was stirred at 80 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the mixture is cooled to 25 ℃, and the mixture is concentrated under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a gradient of 0% → 10% methanol/dichloromethane and the fractions were stripped of solvent by rotary evaporation under reduced pressure to give compound 20-3 (yellow oil, 1.2 g, 61% yield). MS (ESI, m/z) 396.2[ m + H ], [ solution ] of] ⁺ ； ¹ HNMR(400MHz,CDCl ₃ )δ4.22(q,J＝7.1Hz,2H),4.15(s,2H),3.76–3.59(m,20H),2.70–2.60(m,4H),2.32(s,3H),1.75–1.66(m,2H),1.29(t,J＝7.1Hz,3H)。

Step 4

Under nitrogen protection at 25 degrees celsius, the nitrogen is added to a mixture of 1-7 (0.5 g, 0.9 mmol,1.0 equiv) of N-methylpyrrolidinone (20.0 ml) was added 20-3 (0.5 g, 1.2 mmol, 1.3 equiv), cesium fluoride (0.2 g, 1.4 mmol, 1.5 equiv) and N, N-diisopropylethylamine (0.3 g, 1.9 mmol, 2.0 equiv). The reaction solution was reacted at 120 ℃ for 2 hours, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was cooled to 25 ℃. Concentrating under reduced pressure to remove excessive reagent to obtain crude product. The crude product was purified by column chromatography on silica gel, the mobile phase was eluted with a gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 20-4 (yellow oil, 463 mg, 56% yield). MS (ESI, m/z) 838.1/840.1/842.1M + H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.72–7.69(m,1H),4.49(t,J＝6.5Hz,2H),4.21(q,J＝7.2Hz,2H),4.14(s,2H),3.80–3.71(m,6H),3.71–3.67(m,2H),3.66–3.58(m,18H),2.67(s,3H),2.50–2.25(m,4H),2.12–1.97(m,2H),1.49(s,9H),1.28(t,J＝7.1Hz,3H)。

Step 5

20-4 (160.0 mg, 0.2 mmol, 1.0 equiv.) was dissolved in a mixed solution of tetrahydrofuran/methanol/water (1/1, 5.0 ml) with stirring at 25 deg.C, and lithium hydroxide (16.0 mg, 0.4 mmol, 2.0 equiv.) was added. The mixture was allowed to react at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, 1.0 mol/l of dilute hydrochloric acid was added to the reaction solution to adjust the pH to 5. Concentrating under reduced pressure to obtain intermediate. The intermediate was then dissolved in N, N-dimethylformamide (5.0 ml) with stirring at 25 ℃ and 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (90.0 mg, 0.24 mmol, 1.2 eq) was added. The reaction mixture was reacted at 25 ℃ for 15 minutes, and N, N-diisopropylethylamine (91.8 mg, 0.7 mmol, 3.6 equivalents) and (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) -amide were added thereto]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thia-ne)Azol-5-yl) phenyl]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (105.2 mg, 0.2 mmol, 1.2 eq). The reaction solution was reacted at 25 ℃ for 2 hours, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished. Concentrating under reduced pressure to obtain crude product. The crude product was purified by reverse phase chromatography (C18 column) eluting with 30% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 25 minutes; detector, UV254/220 nm; compound 20-5 was obtained (white solid, 180.0 mg, 76% yield). MS (ESI, m/z) 1236.3/1238.3/1240.3[ 2 ] M + H ] ⁺ 。

Step 6

To a mixed solution of compound 20-5 (162.3 mg, 0.1 mmol, 1.0 eq) in 1, 4-dioxane/water (1/1, 6.0 ml) was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (53.1 mg, 0.2 mmol, 1.5 eq) and [1,1' -bis (diphenylphosphino) ferrocene ] under nitrogen protection at 25 deg.C]Palladium dichloride dichloromethane complex (10.7 mg, 0.01 mmol, 0.1 eq) and sodium carbonate (27.8 mg, 0.3 mmol, 2.0 eq). The mixture was stirred at 90 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the mixture was cooled to room temperature and concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 30 minutes; detector UV 254/220 nm; compound 20-6 was obtained (white solid, 140.0 mg, 81% yield). MS (ESI, m/z) 1300.5/1302.5[ m + H ]] ⁺ ； ¹ HNMR(400MHz,CDCl ₃ )δ8.67(s,1H),7.80–7.81(m,2H),7.46–7.27(m,9H),7.25–7.19(m,1H),7.17–7.13(m,1H),5.13–5.03(m,1H),4.79–4.68(m,1H),4.63–4.54(m,2H),4.12–3.94(m,3H),3.91–3.76(m,3H),3.76–3.52(m,22H),3.49(s,9H),2.96–2.66(m,3H),2.51(s,3H),2.49–2.36(m,2H),2.20–2.06(m,3H),1.51(s,9H),1.48–1.45(m,3H),1.08–1.02(m,8H)。

Step 7

Chiral resolution of the compound 20-6 (140 mg) obtained in step 6 by preparative chiral high performance liquid chromatography: a chiral column NB-Lux 5 mi-Cellulose-5, 2.12x25 cm, 5 microns; mobile phase A: n-hexane/dichloromethane =3/1 (0.1% diethylamine), mobile phase B: ethanol; flow rate: 15 ml/min; elution with 50% mobile phase B for 28.2 min; detector UV220/254 nm, resulting in two products. The product with shorter retention time (11.2 min) was compound 20-6a (white solid, 60.0 mg, 43% recovery), compound 20-6a: MS (ESI, m/z) 1300.5/1302.5[ 2 ] M + H ] ⁺ (ii) a The product at longer retention time (20.6 min) was compound 20-6b (white solid, 61.0 mg, 44% recovery), compound 20-6b: MS (ESI, m/z) 1300.5/1302.5[ m + H ]] ⁺ 。

Step 8

To a solution of compound 20-6a (60.0 mg, 0.05 mmol, 1.0 eq) in dichloromethane (2.0 ml) was added trifluoroacetic acid (1.0 ml) with stirring at zero degrees centigrade. The mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: gemini-NX C18 AXAI Packed,21.2x150 mm, 5 microns; a mobile phase A: water (0.1% ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with 10% B phase over 2 minutes, followed by 10% → 28% B phase gradient over 2.5 minutes, and finally 28% → 58% B phase gradient over 11.5 minutes; detector UV 254/220 nm; compound 20a was obtained (white solid, 23.0 mg, 42% yield). MS (ESI, m/z) 1200.4/1202.4[ m ] +H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.02(s,1H),8.98(s,1H),8.44(d,J＝7.7Hz,1H),7.94–7.90(m,1H),7.83–7.78(m,1H),7.49–7.40(m,3H),7.40–7.32(m,3H),7.28(d,J＝2.4Hz,1H),7.24–7.18(m,2H),7.08–7.04(m,1H),5.20–5.10(m,1H),4.96–4.86(m,1H),4.54(d,J＝9.5Hz,1H),4.48–4.40(m,1H),4.39–4.32(m,2H),4.31–4.25(m,1H),3.97–3.93(m,2H),3.81–3.73(m,4H),3.65–3.42(m,21H),2.94–2.84(m,4H),2.49–2.46(m,3H),2.45(s,3H),2.18(s,3H),2.09–2.00(m,1H),1.92–1.82(m,2H),1.81–1.72(m,1H),1.42–1.32(m,3H),0.93(s,9H)。

Step 8'

To a solution of compound 20-6b (61.0 mg, 0.05 mmol, 1.0 eq) in dichloromethane (2.0 ml) was added trifluoroacetic acid (1.0 ml) with stirring at zero degrees centigrade. The mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: gemini-NX C18 AXAI Packed,21.2x150 mm, 5 microns; a mobile phase A: water (0.1% ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a gradient of phase B of 26% → 55% over 10 minutes; detector UV 254/220 nm; yield 20b (white solid, 24.0 mg, 42% yield). MS (ESI, m/z) 1200.4/1202.4[ m ] +H ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.02(s,1H),8.98(s,1H),8.44(d,J＝7.7Hz,1H),7.94–7.90(m,1H),7.83–7.78(m,1H),7.49–7.40(m,3H),7.40–7.32(m,3H),7.28(d,J＝2.4Hz,1H),7.24–7.18(m,2H),7.08–7.04(m,1H),5.20–5.10(m,1H),4.96–4.86(m,1H),4.54(d,J＝9.5Hz,1H),4.48–4.40(m,1H),4.39–4.32(m,2H),4.30–4.25(m,1H),3.97–3.93(m,2H),3.82–3.74(m,4H),3.65–3.42(m,21H),2.94–2.86(m,4H),2.49–2.46(m,3H),2.45(s,3H),2.19(s,3H),2.09–2.00(m,1H),1.93–1.82(m,2H),1.81–1.72(m,1H),1.41–1.33(m,3H),0.93(s,9H)。

Example 21

(2s, 4r) -1- ((S) -18- (tert-butyl) -2- (1- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) azetidin-3-yl) -16-oxo-5, 8,11, 14-tetraoxo-2, 17-diazacyclodecane-19-acyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 21a; (2S, 4R) -1- ((S) -18- (tert-butyl) -2- (1- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) azetidin-3-yl) -16-oxo-5, 8,11, 14-tetraoxo-2, 17-diazacyclodecane-19-acyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 21b.

Step 1

To a solution of compound tert-butyl 3- (methylamino) azetidine-1-carboxylate (3.00 g, 15.302 mmol, 1.00 eq) in acetonitrile (30 ml) was added N, N-diisopropylethylamine (3.96 g, 30.604 mmol, 2.00 eq) and chloropropene (1.41 g, 18.362 mmol, 1.20 eq) with stirring at 25 ℃. The reaction solution was stirred at 80 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature. The solvent was removed by concentration under reduced pressure, water (30 ml) was added, and then extracted with ethyl acetate (30 ml × 3). After the combination, the organic layers were washed with 30 ml of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was eluted with a 0% → 25% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 21-1 (red oil, 3 g, 86% yield). MS (ESI, m/z) 227.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ6.37–6.23(m,1H),5.71–5.60(m,2H),4.43–4.34(m,2H),4.34–4.24(m,2H),3.74–3.62(m,1H),3.43–3.37(m,2H),2.61(s,3H),1.90(s,9H)。

Step 2

To a solution of compound 21-1 (424.21 mg, 1.781 mmol, 1.50 eq) in dichloromethane (3 ml) was added trifluoroacetic acid (1 ml) with stirring at 25 ℃. The reaction solution was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the reaction liquid is decompressed and concentrated to obtain an intermediate crude product. This intermediate was dissolved in N-methylpyrrolidone (5 ml), to which 1-7 (600.00 mg, 1.187 mmol, 1.00 eq) and N, N-diisopropylethylamine (1.62 g, 11.870 mmol, 10.00 eq) were subsequently added. The resulting mixture was stirred at 60 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was purified by reverse phase flash chromatography (C18 column) and eluted with 10% → 95% acetonitrile/water mobile phase (0.1% formic acid) within 20 minutes; detector UV254 nm; compound 21-2 was obtained as a pale yellow solid (650 mg, 96% yield). MS (ESI, m/z) 569.3/571.3/573.3[ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.56(d,J＝1.9Hz,1H),5.95–5.81(m,1H),5.29–5.15(m,2H),4.31–4.18(m,2H),4.16–4.03(m,2H),3.62(s,8H),3.49–3.37(m,1H),3.10–2.95(m,2H),2.24(s,3H),1.49(s,9H)。

Step 3

To a solution of compound 21-2 (570.0 mg, 0.95 mmol, 1.00 eq) in dichloromethane (10 ml) was added 1, 3-dimethylbarbituric acid (234.3 mg, 1.43 mmol, 1.50 eq), tetrakis (triphenylphosphine) palladium (57.8 mg, 0.048 mmol, 0.05 eq) with stirring at 25 ℃. The mixture is at 25 deg.C The reaction was stirred for 4 hours at ambient conditions and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was eluted with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 21-3 (light yellow solid, 350 mg, 88% yield). MS (ESI, m/z) 529.1/531.1/532 [ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.57–7.54(m,1H),4.42–4.33(m,2H),3.98–3.86(m,2H),3.74–3.68(m,1H),3.62(s,8H),2.46(s,3H),1.49(s,9H)。

Step 4

Trifluoroacetic acid (5.0 ml) was added dropwise to a solution of tert-butyl acetate-tetraethylene glycol (623 mg, 2.0 mmol, 1 eq) in dichloromethane (5.0 ml) with stirring at zero degrees centigrade. The resulting mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. Toluene (15.0 ml) was added to the crude product, and the residue of trifluoroacetic acid was further removed by concentration under reduced pressure, and this operation was repeated 3 times to obtain a crude product of compound 21-4 (pale yellow oil, 490 mg). The crude product was used directly in the next reaction. MS (ESI, m/z) 253.1[ 2 ], M + H] ⁺ 。

Step 5

To a solution of compound 21-4 (490.00 mg, 1.9 mmol, 1.00 eq) in N, N-dimethylformamide (5.0 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (887.2 mg, 2.3 mmol, 1.20 eq) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 0.5 hour. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) was added to the above-mentioned reaction mixture ]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] amide]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (1.03 g, 2.1 mmol, 1.10 equivalents) and N, N-diisopropylethylamine (1.23 g, 9.5 mmol, 5.00 equivalents). The mixture was allowed to react at 25 ℃ for a further 3 hours, the course of which was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was purified by reverse phase flash chromatography (C18 column) and eluted with acetonitrile/water mobile phase of 5% → 95% in 14 minutes; detector UV254 nm; compound 21-5 was obtained (light yellow solid, 785 mg, 60% yield). MS (ESI, m/z) 679.3[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.99(s,1H),8.45(d,J＝7.7Hz,1H),7.46–7.36(m,5H),4.96–4.85(m,1H),4.59–4.53(m,1H),4.49–4.42(m,1H),4.32–4.27(m,1H),3.99–3.94(m,2H),3.65–3.47(m,16H),3.44–3.40(m,2H),2.46(s,3H),2.11–2.01(m,1H),1.84–1.73(m,1H),1.46–1.33(m,3H),0.95(s,9H)。

Step 6

To a solution of compound 21-5 (785.00 mg, 1.16 mmol, 1.00 eq) in acetonitrile (8.0 ml) was added 2-iodoxybenzoic acid (356.6 mg, 1.27 mmol, 1.10 eq) with stirring at 25 ℃. The mixture was reacted at 60 ℃ for 2.5 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 8% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 21-6 (white solid, 615.3 mg, 78% yield). MS (ESI, m/z) 677.2[ 2 ], [ M ] +H ] ⁺ 。

Step 7

To compound 21-6 (332.13 mg, 0.49 mmol, 1.30 equiv.) and compound 21-2 (200.00 mg, 0) were added with stirring at 25 deg.C38 mmol, 1.00 eq) in methanol (8 ml) was added acetic acid (22.67 mg, 0.38 mmol, 1.00 eq) and sodium cyanoborohydride (35.58 mg, 0.57 mmol, 1.50 eq). The mixture was reacted at 25 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by preparative thin layer chromatography (dichloromethane/methanol = 12/1) to give compound 21-7 (white solid, 356 mg, 78% yield). MS (ESI, m/z) 1189.4/1191.3/1193.4[ 2 ] M + H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ8.98(s,1H),8.43(d,J＝7.8Hz,1H),7.77(s,1H),7.46–7.34(m,5H),5.13(d,J＝3.5Hz,1H),4.96–4.85(m,1H),4.54(d,J＝9.5Hz,1H),4.44(t,J＝8.1Hz,1H),4.33–4.24(m,1H),4.19–4.07(m,2H),3.99–3.93(m,2H),3.92–3.84(m,2H),3.71–3.46(m,26H),2.45(s,3H),2.26–2.15(m,3H),2.11–2.01(m,1H),1.84–1.71(m,1H),1.43(s,9H),1.37(d,J＝6.9Hz,3H),0.94(s,9H)。

Step 8

To a solution of compounds 21-7 (200.00 mg, 0.160 mmol, 1.00 eq) in tetrahydrofuran/water (10/1, 2.2 ml) was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (68.06 mg, 0.252 mmol, 1.5 eq), potassium phosphate (71.31 mg, 0.336 mmol, 2.0 eq) and chlorine (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl ] at 25 ℃ with stirring under nitrogen protection ]Palladium (II) (14.22 mg, 0.017 mmol, 0.1 eq). The reaction solution was stirred at 60 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, cooling the reaction liquid to 25 ℃, and concentrating under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 21-8 (off-white solid, 170 mg, 84% yield). MS (ESI, m/z) 1253.7/1255.7[ m ] +H] ⁺ 。

Step 9

Chiral resolution of the compound 20-8 (170 mg) obtained in step 8 by preparative chiral high performance liquid chromatography: a chiral column NB-Lux 5 mi-Cellulose-5, 2.12x25 cm, 5 microns; mobile phase A: n-hexane/methyl tert-butyl ether =1/1 (0.5%, 2 mol/l methanolic ammonia), mobile phase B: ethanol; flow rate: 20 ml/min; elution was performed with 50% phase B over 8 minutes with detector UV225/250 nm to give two products. The product with shorter retention time (8.645 minutes) was compound 21-8a (off-white solid, 50 mg, 29% recovery), compound 21-8a: MS (ESI, m/z) 1253.7/1255.7[ 2 ], [ M ] +H] ⁺ (ii) a The product with longer retention time (15.475 min) was compound 21-8b (off-white solid, 50 mg, 29% recovery), compound 21-8b: MS (ESI, m/z) 1253.7/1255.7[ 2 ], [ M ] +H ] ⁺ 。

Step 10

To a solution of compound 21-8a (50.00 mg, 0.040 mmol, 1.00 eq) in dichloromethane (3 ml) was added trifluoroacetic acid (1 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 0.5 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; the gradient eluted with a 5% → 25% B phase gradient over 7 minutes; detector UV254/220 nm; yield 21a (off-white solid, 10 mg, 21% yield). MS (ESI, m/z) 1153.5/1155.4[ m + H ], [] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.90–8.86(m,1H),7.81–7.73(m,2H),7.47–7.38(m,5H),7.30–7.17(m,3H),7.03(d,J＝2.4Hz,1H),5.06–4.97(m,1H),4.69(s,1H),4.63–4.54(m,1H),4.44(s,1H),4.34–4.22(m,2H),4.14–4.02(m,4H),3.91–3.43(m,22H),3.14–3.04(m,4H),2.71–2.62(m,2H),2.48(s,3H),2.32(s,3H),2.26–2.14(m,1H),2.02–1.91(m,1H),1.60–1.48(m,3H),1.40–1.26(m,2H),1.04(s,9H),0.96–0.84(m,1H)。

Step 10'

To a solution of compound 21-8b (50.00 mg, 0.040 mmol, 1.00 eq.) in dichloromethane (3 ml) was added trifluoroacetic acid (1 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 0.5 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X250 mm, 5 μm; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient eluted with 5% → 25% b phase gradient in 7 min; detector UV254/220 nm; yield 21b (off-white solid, 12 mg, 25% yield). MS (ESI, m/z) 1153.5/1155.4[ m + H ], [ ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.00(s,1H),8.98(s,1H),8.44(d,J＝7.7Hz,1H),7.79(d,J＝8.3Hz,1H),7.74(d,J＝1.6Hz,1H),7.48–7.40(m,3H),7.40–7.32(m,3H),7.26(d,J＝2.4Hz,1H),7.21(d,J＝4.0Hz,2H),7.03(d,J＝2.3Hz,1H),5.22–5.08(m,1H),4.97–4.83(m,1H),4.54(d,J＝9.5Hz,1H),4.50–4.39(m,1H),4.34–4.22(m,1H),4.18–4.05(m,2H),3.98–3.83(m,4H),3.72–3.63(m,4H),3.62–3.44(m,18H),2.96–2.88(m,4H),2.45(s,3H),2.18(s,3H),2.09–1.99(m,1H),1.82–1.73(m,1H),1.37(d,J＝7.0Hz,3H),1.29–1.21(m,1H),0.93(s,9H)。

Example 22

(2S, 4R) -1- ((S) -2- (5- (4- (5- ((3- ((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) pentyl) piperazin-1-yl) pentanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide trihydrochloride 22.

Step 1

To a 100 ml round bottom flask was added methyl 5-bromovalerate (2.0 g, 9.7 mmol, 1.0 eq), acetonitrile (30.0 ml), 1-tert-butoxycarbonylpiperazine (2.1 g, 10.7 mmol, 1.1 eq) and potassium carbonate (2.8 g, 19.4 mmol, 2.0 eq) sequentially with stirring at 25 ℃. The resulting mixture was stirred at 85 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, cooling the reaction liquid to room temperature, filtering, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and give compound 22-1 (yellow liquid, 2.8 g, 91% yield). MS (ESI, m/z) 301.2[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ3.68(s,3H),3.47–3.40(m,4H),2.40–2.31(m,8H),1.73–1.60(m,2H),1.56–1.52(m,2H),1.47(s,9H)。

Step 2

To a solution of compound 22-1 (1.4 g, 4.7 mmol, 1.0 eq) in methanol/tetrahydrofuran/water (1/1/1, 13.5 ml) was added lithium hydroxide monohydrate (0.3 g, 7.0 mmol, 1.5 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour. The reaction process is carried out by liquid chromatography and thin layer chromatographyAnd (5) monitoring. After completion of the reaction, a hydrochloric acid solution (1 mol per liter, 7 ml) was added dropwise to the reaction solution at zero degrees centigrade. The reaction was concentrated to give 22-2 (white solid, 1.8 g) as a crude product. The crude product was used directly in the next synthesis. MS (ESI, m/z) 287.2, [ 2 ], [ M ] +H] ⁺ 。

Step 3

To a solution of compound 22-2 (472.2 mg) in N, N-dimethylformamide (5.0 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (513.1 mg, 1.3 mmol, 1.2 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 15 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) was added to the reaction system]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] amide]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (500.0 mg, 1.1 mmol, 1.0 eq) and N, N-diisopropylethylamine (0.6 ml, 3.3 mmol, 3 eq). The resulting mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was directly purified by a reverse phase flash column (C18 column), and eluted with 25% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 25 minutes; detector UV254 nm; compound 22-3 was obtained (white solid, 640 mg, 83% yield). MS (ESI, m/z) 713.4[ 2 ], [ M ] +H ] ⁺ 。

Step 4

To a solution of compound 22-3 (300.0 mg, 0.4 mmol, 1.0 eq) in dichloromethane (6.0 ml) was added trifluoroacetic acid (2.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product 22-4 is obtained by decompression and concentration(yellow solid, 410.0 mg). The compound was used in the next synthesis without further purification. MS (ESI, m/z) 613.2[ 2 ], [ M ] +H] ⁺ 。

Step 5

To a solution of compound 22-4 (72.9 mg, 0.1 mmol, 1.2 eq), sodium acetate (12.2 mg, 0.15 mmol, 1.5 eq), acetic acid (8.9 mg, 0.15 mmol, 1.5 eq) and sodium cyanoborohydride (18.7 mg, 0.3 mmol, 3.0 eq) in methanol (2.0 ml) was added dropwise compound 3-6a (70.0 mg, 0.1 mmol, 1.0 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 22-5a (pale yellow solid, 70 mg, yield 54%) was obtained. MS (ESI, m/z) 1302.8/1304.8[ 2[ M ] +H ] ⁺ 。

Step 6

To a solution of compound 22-5a (50.0 mg, 0.04 mmol, 1.0 eq) in methanol (2.0 ml) was added a solution of 1, 4-dioxane hydrochloric acid (4 mol/l, 2.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X150 mm, 5 μm; mobile phase A: water (0.05% hydrochloric acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 28% B phase gradient over 7 minutes; detector UV 254/220 nm; compound 22 was obtained (yellow solid, 21.5 mg, 45% yield). MS (ESI, m/z): 1202.6/1208.6[ M ] +H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ12.00(s,2H),10.79(s,1H),10.08(d,J＝10.0Hz,1H),9.84(s,1H),9.10(s,1H),8.46(d,J＝7.8Hz,1H),8.00(d,J＝1.7Hz,1H),7.91(d,J＝9.0Hz,1H),7.82(d,J＝8.3Hz,1H),7.54–7.35(m,5H),7.32(d,J＝2.4Hz,1H),7.22(d,J＝6.1Hz,2H),7.12(d,J＝2.3Hz,1H),4.92(t,J＝7.2Hz,2H),4.52–4.40(m,5H),4.29(s,1H),4.17(s,2H),3.99–3.83(m,2H),3.75–3.72(m,3H),3.63–3.58(m,3H),3.49(d,J＝9.6Hz,4H),3.27–3.23(m,2H),3.15–3.05(m,6H),2.75(d,J＝4.8Hz,3H),2.47(s,3H),2.25(d,J＝9.1Hz,5H),1.98–1.92(m,5H),1.79–1.71(m,7H),1.56(d,J＝7.3Hz,2H),1.39–1.30(m,5H),0.95(s,9H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-122.00。

Example 23

(2s, 4r) -1- ((S) -2- (14- ((3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) tetradecanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 23a; (2S, 4R) -1- ((S) -2- (14- ((3- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) tetradecanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 23b.

Step 1

To a solution of 14-hydroxymyristic acid (200.0 mg, 0.818 mmol, 1.0 eq) in N, N-dimethylformamide (5 ml) was added 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (373.42 mg, 0.9 eq) with stirring at 25 deg.C82 mmol, 1.2 equivalents). The resulting mixture was reacted at 25 ℃ for 30 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) was added to the reaction mixture]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (363.86 mg, 0.818 mmol, 1.0 eq) and N, N-diisopropylethylamine (528.42 mg, 4.090 mmol, 5.0 eq). The resulting mixture was allowed to continue to react at 25 deg.C for 2.5 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was directly purified by reverse phase flash chromatography (C18 column) and eluted with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) within 20 minutes; detector UV254 nm; compound 23-1 was obtained (white solid, 320 mg, 58% yield). MS (ESI, m/z) 669.4[ M-H ], [ m-H ]] ^- ； ¹ H NMR(300MHz,DMSO-d ₆ )δ8.99(s,1H),8.38(d,J＝7.8Hz,1H),7.79(d,J＝9.3Hz,1H),7.47–7.35(m,4H),5.15–5.08(m,1H),5.00–4.85(m,1H),4.52(d,J＝9.3Hz,1H),4.48–4.38(m,1H),4.38–4.22(m,2H),3.66–3.55(m,2H),2.46(s,3H),2.31–2.19(m,1H),2.15–1.95(m,2H),1.85–1.74(m,1H),1.51–1.35(m,7H),1.29–1.20(m,19H),0.93(s,9H)。

Step 2

Compound 23-1 (300 mg, 0.425 mmol, 1.0 eq), 2-iodoxybenzoic acid (125.2 mg, 0.425 mmol, 1.0 eq) and acetonitrile (6 ml) were added sequentially to the reaction flask at 25 ℃ with stirring under nitrogen. The mixture was stirred at 60 ℃ under nitrogen for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is cooled to room temperature, and the reaction liquid is concentrated under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 12% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and give compound 23-2 (white solid, 174 mg, 58% yield). MS (ESI, m/z) 669.4[ 2 ] M + H ] ⁺ 。

Step 3

To a 25 ml single neck flask were added 2-9a (30 mg, 0.048 mmol, 1.0 eq), 23-2 (37 mg, 0.055 mmol, 1.1 eq), acetic acid (4.53 mg, 0.075 mmol, 1.5 eq), methanol (4 ml) and sodium cyanoborohydride (4.74 mg, 0.075 mmol, 1.5 eq) sequentially with stirring at zero degrees centigrade under nitrogen protection. The resulting mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 23-3a (white solid, 44.5 mg, 74% yield). MS (ESI, m/z) 1248.6/1250.6[ 2 ], [ M + H ]] ⁺ 。

Step 4

Compound 23-3a (44.5 mg, 0.035 mmol, 1.0 eq), dichloromethane (4 ml) and trifluoroacetic acid (2 ml) were added sequentially to a 25 ml single-neck flask with stirring at zero degrees celsius. The reaction solution was stirred at 20 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, concentrating the reaction solution to obtain a crude product, purifying the crude product by a reverse phase flash chromatography column (C18 column), and eluting the crude product by using a 5% → 95% acetonitrile/water mobile phase (0.05% ammonia water) within 20 minutes; detector UV 254 nm; compound 23a was obtained (white solid, 5 mg, 12% yield). MS (ESI, m/z) 1148.6/1150.6[ M + H ] ] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.87(s,1H),7.95–7.92(m,1H),7.74(d,J＝8.2Hz,1H),7.45–7.36(m,5H),7.27–7.16(m,3H),7.04–7.01(m,1H),5.04–4.95(m,1H),4.61(s,1H),4.59–4.53(m,1H),4.53–4.47(m,2H),4.44–4.39(m,1H),3.96–3.85(m,5H),3.77–3.71(m,1H),3.10–3.02(m,4H),2.77–2.68(m,2H),2.54–2.45(m,5H),2.37(s,3H),2.32–2.15(m,3H),2.09–2.01(m,2H),1.99–1.90(m,1H),1.62–1.47(m,7H),1.37–1.33(m,1H),1.30–1.21(m,16H),1.06–0.99(m,9H),0.92–0.81(m,1H)。

Step 3'

To a 25 ml single neck flask were added 2-9b (40 mg, 0.067 mmol, 1.0 eq), 23-2 (58 mg, 0.087 mmol, 1.3 eq), acetic acid (6.04 mg, 0.101 mmol, 1.5 eq), methanol (5 ml) and sodium cyanoborohydride (6.33 mg, 0.101 mmol, 1.5 eq) sequentially with stirring at zero degrees centigrade under nitrogen protection. The resulting mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 23-3b (white solid, 71 mg, 85% yield). MS (ESI, m/z) 1248.6/1250.6[ 2 ], [ M + H ]] ⁺ 。

Step 4'

To a 25 ml single-neck flask were added 23-3b (71 mg, 0.058 mmol, 1.0 eq), dichloromethane (4 ml) and trifluoroacetic acid (2 ml) sequentially with stirring at zero degrees centigrade. The reaction solution was stirred at 20 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by a reverse phase flash column (C18 column) and eluted with 5% → 95% acetonitrile/water mobile phase (0.05% ammonia) within 20 minutes; detector UV 254 nm; compound 23b was obtained (white solid, 15 mg, 23% yield). MS (ESI) ,m/z):1148.6/1150.6[M+H] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.86(s,1H),7.95–7.90(m,1H),7.74(d,J＝8.3Hz,1H),7.47–7.33(m,5H),7.26–7.15(m,3H),7.05–7.01(m,1H),5.03–4.95(m,1H),4.61(s,1H),4.58–4.53(m,1H),4.53–4.47(m,2H),4.44–4.39(m,1H),3.95–3.84(m,5H),3.77–3.70(m,1H),3.09–3.01(m,4H),2.73–2.63(m,2H),2.51–2.42(m,5H),2.34–2.30(m,3H),2.29–2.15(m,3H),2.08–1.99(m,2H),1.98–1.89(m,1H),1.64–1.45(m,7H),1.37–1.33(m,1H),1.30–1.21(m,16H),1.02(d,J＝11.0Hz,9H),0.94–0.83(m,1H)。

Example 24

(2s, 4r) -1- ((S) -2- (4- (4- (4- ((3- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyl) piperazin-1-yl) butyramido) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide dicarboxylate 24.

Step 1

To a solution of methyl 4-chlorobutyrate (1.10 g, 8.1 mmol, 1.00 eq) in acetonitrile (10 ml) was added 1-tert-butoxycarbonylpiperazine (1.50 g, 8.1 mmol, 1.00 eq) and potassium carbonate (2.23 g, 16.2 mmol, 2.00 eq) with stirring at 25 ℃. The reaction solution was stirred at 80 ℃ for 4 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the reaction solution is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 5% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and give compound 24-1 (white solid, 623.4 mg, 26% yield). MS (ESI, m/z) 287.1[ deg. ] M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ3.69(s,3H),3.43(t,J＝5.1Hz,4H),2.43–2.34(m,8H),1.90–1.78(m,2H),1.48(s,9H)。

Step 2

To a solution of compound 24-1 (619.0 mg, 2.16 mmol, 1.0 equiv) in methanol/tetrahydrofuran/water (1/1, 7.5 ml) was added lithium hydroxide monohydrate (136.0 mg, 3.24 mmol, 1.5 equiv) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, a hydrochloric acid solution (1 mol per liter, 3.3 ml) was added dropwise to the reaction solution at zero degrees centigrade. The reaction was concentrated to give crude 24-2 (white solid, 938 mg, crude). The crude product was used directly in the next synthesis. MS (ESI, m/z) 273.2[ 2 ], [ M ] +H ] ⁺ 。

Step 3

To a solution of 24-2 (320.00 mg, crude) in N, N-dimethylformamide was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (491.44 mg, 1.228 mmol, 1.10 equiv) with stirring at 25 ℃. The mixture was stirred at 25 ℃ for 1 hour. N, N-diisopropylethylamine (607.43 mg, 4.465 mmol, 4.00 eq.) and (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) were then added to the reaction mixture]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (10.45 mg, 0.022 mmol, 1.1 eq). The mixture was stirred at 25 ℃ for an additional 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was directly purified by a reverse phase flash column (C18 column), and eluted with 10% → 70% acetonitrile/water mobile phase (0.1% ammonia) within 20 minutes; detector UV254 nm; compound 24-3 was obtained (a bright yellow solid,560 mg, 71% yield). MS (ESI, m/z) 699.5[ 2 ], [ M + H ]] ⁺ 。

Step 4

To an 8 ml reaction flask were added 24-3 (50.0 mg, 0.068 mmol, 1.00 eq), ethyl acetate (1.00 ml), and a 1, 4-dioxane solution of hydrochloric acid (4 mol/l, 1.00 ml) sequentially with stirring at zero degrees centigrade. The resulting mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, concentration was performed under reduced pressure to obtain a crude product of compound 24-4 (bright yellow solid, 40 mg, crude product). The crude product was used directly in the next reaction. MS (ESI, m/z) 599.3[ m + H ] ] ⁺ 。

Step 5

To an 8 ml reaction flask were added, under stirring at 25 ℃, compound 24-4 (30.56 mg, 0.048 mmol, 0.85 eq), methanol (2.00 ml, 62.418 mmol, 822.71 eq), acetic acid (7.21 mg, 0.114 mmol, 2.00 eq), sodium cyanoborohydride (7.55 mg, 0.114 mmol, 2.00 eq), and compound 8-6a (40.00 mg, 0.057 mmol, 1.00 eq), in that order. The mixture was reacted for 2 hours at 25 ℃ with stirring. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% methanol/water mobile phase (0.1% ammonia) over 20 minutes; detector UV254 nm; compound 24-5a was obtained (white solid, 50 mg, 70% yield). MS (ESI, m/z) 1148.6/1150.6[ 2 ] M + H] ⁺ 。

Step 6

To an 8 ml reaction flask were added sequentially compound 24-5a (25.0 mg, 0.019 mmol, 1.0 eq), dichloromethane (4.0 ml) and trifluoroacetic acid (1.0 ml) with stirring at zero degrees centigrade. The resulting mixture was stirred at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% formic acid) over 20 minutes; detector UV254 nm; compound 24 was obtained (white solid, 7.2 mg, 30% yield). MS (ESI, m/z) 1149.6[ m + H ] ] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.77(s,1H),8.33(s,2H),7.92(d,J＝1.7Hz,1H),7.69–7.64(m,1H),7.35–7.24(m,5H),7.18(d,J＝2.4Hz,1H),7.13–7.07(m,2H),6.93(d,J＝2.4Hz,1H),4.94–4.86(m,1H),4.73–4.65(m,1H),4.55–4.41(m,4H),4.37–4.31(m,1H),4.05–3.93(m,4H),3.81–3.75(m,1H),3.68–3.61(m,1H),3.35–3.26(m,4H),3.24–3.22(m,1H),3.19–3.14(m,1H),3.06–2.98(m,2H),2.81–2.63(m,10H),2.61–2.55(m,2H),2.53–2.47(m,2H),2.39–2.35(m,3H),2.31–2.25(m,2H),2.22–2.07(m,3H),1.90–1.81(m,1H),1.81–1.60(m,4H),1.57–1.50(m,2H),1.47–1.37(m,3H),0.93(d,J＝9.5Hz,9H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-123.10。

Example 25

(2S, 4R) -1- ((S) -2- (3- (3- ((S) -2- ((((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) propoxy) propylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide dihydrochloride 25.

Step 1

To a solution of 3-buten-1-ol (10.0 g, 131.7 mmol, 1.0 eq), triethylamine (28.0 g, 263.55 mmol, 2.0 eq) and 4-dimethylaminopyridine (2.5 g, 19.7 mmol, 0.15 eq) in dichloromethane (100 ml) was added dropwise p-toluenesulfonyl chloride (29.1 g, 144.9 mmol, 1.1 eq) at zero degrees centigrade, with stirring under nitrogen. After the dropwise addition, the mixture was reacted for 16 hours at 25 ℃ under nitrogen protection with stirring. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, 200 ml of water was added to the system to quench the reaction. The mixture was extracted with dichloromethane (200 ml x 3), the organic phases were combined, the organic phases were dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 25% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 25-1 (colorless oil, 29.00 g, 92.4% yield). MS (ESI, m/z) 227.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.82–7.76(m,2H),7.35(d,J＝8.0Hz,2H),5.72–5.62(m,1H),5.10–5.05(m,2H),4.08–4.05(m,2H),2.45(s,3H),2.43–2.37(m,2H)。

Step 2

Compound 25-1 (10.0 g, 132.6 mmol, 2.0 eq) and potassium hydroxide (7.44 g, 132.6 mmol, 2.0 eq) were sequentially added to a 100 ml one-neck flask at 25 ℃ with stirring under nitrogen, and the mixture was heated to 75 ℃ for 30 minutes. Then, 1, 3-propanediol (15.0 g, 62.9 mmol, 1.0 equivalent) was slowly added dropwise to the system at 75 ℃ for not less than 30 minutes, and after the addition was completed, the reaction system was heated to 80 ℃ for 30 minutes to react. The reaction was monitored by thin layer chromatography and after the reaction was complete the reaction system was cooled to 25 ℃. The reaction was quenched with 200 ml of water,extraction was performed with ethyl acetate (200 ml × 3), the organic phases were combined, dried over anhydrous sodium sulfate, and filtered, and the obtained filtrate was subjected to distillation under reduced pressure to remove the solvent. The mobile phase was eluted with a gradient of 0% → 25% ethyl acetate/petroleum ether and the resulting fraction was freed of the solvent by rotary evaporation under reduced pressure, giving compound 25-2 (colorless oil, 2.15 g, 25% yield). ¹ H NMR(400MHz,CDCl ₃ )δ5.86–5.76(m,1H),5.13–5.04(m,2H),3.78–3.75(m,2H),3.65–3.62(m,2H),3.52–3.48(m,2H),2.56(s,1H),2.37–2.31(m,2H),1.86–1.80(m,2H)。

Step 3

To a solution of compound 25-2 (2.15 g, 16.5 mmol, 1.0 eq), triethylamine (3.5 g, 33.1 mmol, 2.0 eq) and 4-dimethylaminopyridine (320 mg, 2.5 mmol, 0.15 eq) in dichloromethane (20 ml) was added p-toluenesulfonyl chloride (3.46 g, 18.1 mmol, 1.1 eq) dropwise at zero degrees centigrade, with stirring under nitrogen. After the dropwise addition, the mixture was reacted for 16 hours at 25 ℃ under nitrogen protection with stirring. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, 20 ml of water was added to the system to quench the reaction. The mixture was extracted with dichloromethane (20 ml x 3), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 25% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 25-3 (colorless oil, 4.0 g, 89% yield). MS (ESI, m/z) 285.1[ m ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.81–7.78(m,2H),7.35(d,J＝8.0Hz,2H),5.79–5.69(m,1H),5.07–4.99(m,2H),4.15–4.11(m,2H),3.45–3.42(m,2H),3.38–3.35(m,2H),2.45(s,3H),2.26–2.20(m,2H),1.92–1.86(m,2H)。

Step 4

To a 100 ml single-neck flask were added sequentially compound 25-3,l-prolinol (0.85 g, 7.9 mmol, 1.2 equivalents), potassium carbonate (1.94 g, 13.3 mmol, 2.0 equivalents) and acetonitrile (20.0 ml) with stirring at 25 ℃. The mixture was reacted at 50 ℃ for 4 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature, insoluble matter was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 5% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 25-4 (white solid, 1.17 g, 77% yield). MS (ESI, m/z) 214.2[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ5.90–5.77(m,1H),5.14–5.02(m,2H),3.66–3.61(m,1H),3.54–3.45(m,4H),3.41–3.37(m,1H),3.22–3.16(m,1H),3.05(s,1H),2.93–2.83(m,1H),2.64–2.58(m,1H),2.39–2.22(m,4H),1.93–1.70(m,6H)。

Step 5

To a solution of compound 16-3a (300.0 mg, 0.465 mmol, 1.0 eq) and 25-4 (125.1 mg, 0.557 mmol, 1.2 eq) in tetrahydrofuran (5.0 ml) was slowly added potassium tert-butoxide (65.8 mg, 0.557 mmol, 1.2 eq) dropwise at zero degrees centigrade, with stirring under nitrogen protection. The mixture was reacted at zero degrees centigrade for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is complete, the mixture is quenched by pouring into 20 ml of water. The mixture was extracted with ethyl acetate (20 ml x 3), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography eluting with a methanol/dichloromethane mobile phase gradient of 0% → 10% and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 25-5a (white solid, 240 mg, 62% yield). MS (ESI, m- z):790.4/792.4[M+H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.84–7.80(m,2H),7.54–7.46(m,2H),7.35–7.30(m,2H),7.19(d,J＝2.4Hz,1H),5.71(s,1H),5.36–5.32(m,2H),5.03–4.96(m,2H),4.41(s,4H),3.95–2.92(m,15H),2.31–1.85(m,13H),1.53(s,9H)。

Step 6

To a 25 ml single-neck flask were added, in order, 25-5a (150.0 mg, 0.18 mmol, 1.0 eq), sodium periodate (166.43 mg, 0.739 mmol, 4.1 eq), ruthenium trichloride (4.28 mg, 0.018 mmol, 0.1 eq), carbon tetrachloride (1.0 ml), acetonitrile (1 ml) and water (1.5 ml) with stirring at 25 ℃. The mixture was allowed to react at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, 10 ml of water is added into the reaction solution for dilution, chloroform/isopropanol (3/1, 10 ml of x 3) is used for extraction, organic phases are combined, the organic phases are dried by anhydrous sodium sulfate, a drying agent is removed by filtration, and the filtrate is concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 20 minutes; detector UV220 nm; compound 25-6a was obtained (white solid, 78 mg, 42% yield). MS (ESI, m/z) 808.4/810.4[ 2 ], [ M + H ]] ⁺ 。

Step 7

To an 8 ml reaction flask were added 25-6a (65.0 mg, 0.064 mmol, 1.0 eq), 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (23.3 mg, 0.077 mmol, 1.2 eq) and N, N-dimethylformamide (1 ml) sequentially at 25 ℃ with stirring. The mixture was reacted at 25 ℃ for 15 minutes. Then (2S,4R) -1- [ (2S) -2- Amino-3, 3-dimethylbutyryl group]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] carbonyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (36.1 mg, 0.077 mmol, 1.2 equivalents) and N, N-diisopropylethylamine (41.5 mg, 0.32 mmol, 5.0 equivalents). The mixture was allowed to react at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, purifying by preparative high performance liquid chromatography, wherein the purification conditions are as follows: XBridge Shield RP18 OBD Column,19x150 mm, 5 microns; mobile phase A: water (0.05% ammonia), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 56% → 79% B phase gradient over 8 minutes, detector UV 254/220 nm; compound 25-7a was obtained (white solid, 60 mg, 72% yield). MS (ESI, m/z) 1220.7/1222.7[ 2 ], [ M + H ]] ⁺ 。

Step 8

To a solution of compound 25-7a (60.0 mg, 0.047 mmol, 1.0 eq) in methanol (2.0 ml) was added a solution of 1, 4-dioxane (4 mol/l, 2.0 ml) of hydrochloric acid with stirring at zero degrees centigrade. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X150 mm, 5 μm; mobile phase A: water (0.05% hydrochloric acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a 5% → 35% B phase gradient over 7 minutes, detector UV 254/220 nm; compound 25 was obtained (light yellow solid, 40 mg, 73% yield). MS (ESI, m/z) 1076.4/1078.5[ m + H ] ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.61(s,1H),10.27–9.60(m,3H),9.06(s,1H),8.67–8.59(m,1H),8.01(d,J＝1.6Hz,1H),7.94(d,J＝9.3Hz,1H),7.81(d,J＝8.3Hz,1H),7.47–7.35(m,5H),7.31(d,J＝2.4Hz,1H),7.26–7.18(m,2H),7.10(d,J＝2.3Hz,1H),4.86–4.75(m,1H),4.74–4.64(m,1H),4.61–4.47(m,3H),4.48–4.30(m,3H),4.29–4.20(m,1H),4.20–4.14(m,2H),4.01–3.88(m,3H),3.67–3.53(m,5H),3.44(t,J＝6.0Hz,3H),3.21–3.08(m,2H),2.44(s,3H),2.41–2.32(m,1H),2.30–2.20(m,1H),2.10–1.84(m,12H),1.01–0.85(m,10H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-121.83。

Example 26

(2s, 4s) -4- (4- ((15- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -12-methyl-3, 6, 9-trioxa-12-azapentadecyl) oxy) phenoxy) -1- ((S) -2-cyclohexyl-2- ((S) -2- (methylamino) propylamino) acetyl) -N- ((R) -1,2,3, 4-tetrahydronaphthalen-1-yl) pyrrolidine-2-carboxamide diformate 26a; (2S, 4S) -4- (4- ((15- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -12-methyl-3, 6, 9-trioxa-12-azapentadecyl) oxy) phenoxy) -1- ((S) -2-cyclohexyl-2- ((S) -2- (methylamino) propylamino) acetyl) -N- ((R) -1,2,3, 4-tetrahydronaphthalen-1-yl) pyrrolidine-2-carboxamide diformate 26b.

Step 1

To a solution of the compound L-cyclohexylglycine methyl ester hydrochloride (10.00 g, 45.739 mmol, 1.00 equiv.) and N-tert-butoxycarbonyl-N-methyl-alanine (9.30 g, 45.739 mmol, 1.00 equiv.) in dichloromethane (100 ml) was added 3- (diethoxyphenoxy) -1,2, 3-benzotriazin-4-one (16.42 g, 54.887 mmol, 1.20 equiv.) and N, N-diisopropylethylamine (20.69 g, 160.087 mmol, 3.50 equiv.) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by reverse phase flash chromatography (C18 column) using 30% → 80% in 20 minutes Eluting with acetonitrile/water mobile phase (0.1% ammonium bicarbonate); detector UV220 nm; compound 26-1 was obtained (pale yellow oil, 9.0 g, 55% yield). MS (ESI, m/z) 357.2[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ6.65(s,1H),4.88–4.60(m,1H),4.54–4.40(m,1H),3.73(s,3H),2.79(s,3H),1.83–1.61(m,5H),1.58–1.52(m,1H),1.50(s,9H),1.33(d,J＝7.1Hz,3H),1.28–1.13(m,2H),1.13–0.92(m,3H)。

Step 2

To a solution of compound 26-1 (9.00 g, 23.986 mmol, 1.00 eq) in tetrahydrofuran/methanol/water (1/1/1, 90 ml) was added lithium hydroxide (906.96 mg, 35.978 mmol, 1.50 eq) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, tetrahydrofuran and methanol were removed by concentration under reduced pressure. Then, 1 mol/l of dilute hydrochloric acid was added to the obtained crude product to adjust the solution pH to 5, and compound 26-2 (white solid, 8.00 g, 77%) was precipitated. MS (ESI, m/z) 343.2[ m ] +H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ12.72(s,1H),7.95–7.39(m,1H),4.82–4.35(m,1H),4.27–4.03(m,1H),2.75(s,3H),1.81–1.50(m,6H),1.38(s,9H),1.30–0.89(m,8H)。

Step 3

To a solution of the compound tetraethylene glycol (5.00 g, 25.743 mmol, 1.00 eq) in tetrahydrofuran (70 ml) was added imidazole (1.31 g, 19.307 mmol, 0.75 eq) and triisopropylchlorosilane (2.48 g, 12.872 mmol, 0.50 eq) with stirring at 25 degrees celsius. The resulting mixture was stirred at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, concentrating under reduced pressure to obtain crude product And (5) preparing the product. The crude product was purified by silica gel column chromatography eluting with a 0% → 55% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 26-3 (red oil, 3 g, 33% yield). MS (ESI, m/z) 351.3[ 2 ] M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ3.83(t,J＝5.6Hz,2H),3.73–3.69(m,2H),3.69–3.62(m,8H),3.61–3.56(m,4H),1.14–1.00(m,21H)。

Step 4

To a solution of (2s, 4r) -1- (tert-butoxycarbonyl) -4-hydroxypyrrolidine-2-carboxylic acid (10.00 g, 41.081 mmol, 1.00 eq) in dichloromethane (150 ml) was added 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (9.12 g, 45.190 mmol, 1.10 eq), 1-hydroxybenzotriazole (7.01 g, 49.298 mmol, 1.20 eq), triethylamine (6.56 g, 61.622 mmol, 1.50 eq) and (R) - (-) -1,2,3, 4-tetrahydro-1-naphthylamine (7.00 g, 45.190 mmol, 1.1 eq) with stirring at zero degrees celsius. The resulting mixture was stirred at 25 ℃ for 5 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 26-4 (white solid, 14 g, 89% yield). MS (ESI, m/z) 361.2[ m + H ] ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.22–6.99(m,4H),5.18–5.01(m,1H),4.54–4.32(m,2H),3.72–3.32(m,2H),2.96–2.69(m,3H),2.50–1.92(m,3H),1.90–1.64(m,3H),1.40(s,9H)。

Step 5

26-4 (4.30 g, 11.333 mmol, 1.00 eq.) was stirred at 25 deg.C under nitrogen protectionTo a solution of 4-benzyloxyphenol (3.11 g, 14.733 mmol, 1.30 eq), tributylphosphine (3.62 g, 16.999 mmol, 1.50 eq) in toluene (50 ml) was added azodicarbonamide (3.08 g, 16.999 mmol, 1.50 eq). The resulting mixture was stirred at 60 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a 0% → 10% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 26-5 (white solid, 4 g, 65% yield). MS (ESI, m/z) 543.3[ m ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.46–7.27(m,6H),7.19–7.12(m,1H),7.12–7.06(m,1H),7.05–6.97(m,1H),6.86–6.80(m,2H),6.61–6.56(m,2H),5.26–5.16(m,1H),5.01(s,2H),4.76–4.70(m,1H),4.46–4.41(m,1H),3.68–3.63(m,2H),2.87–2.71(m,2H),2.71–2.64(m,1H),2.12–2.01(m,1H),1.90–1.74(m,3H),1.67(s,1H),1.45(s,9H)。

Step 6

To a solution of 26-5 (4.00 g, 7.002 mmol, 1.00 eq) in ethyl acetate (40 ml) was added 1, 4-dioxane (4 mol/l, 40.00 ml) of hydrochloric acid with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, concentration under reduced pressure gave compound 26-6 (white solid, 3 g, crude). The crude product was used directly in the next reaction. MS (ESI, m/z) 443.3[ deg. ] M + H ] ⁺ 。

Step 7

26-6 (3.00 g, 6.440 mmol, 1.00 eq.) and 26-2 (2.55 g, 7.084 mmol, 1.10 eq.) of dichloro were added with stirring at 25 deg.CTo a solution of methane (40 ml) was added 3- (diethoxyarthooxyacyloxy) -1,2, 3-benzotriazin-4-one (2.43 g, 7.728 mmol, 1.20 equivalents) and N, N-diisopropylethylamine (3.07 g, 22.539 mmol, 3.50 equivalents). The resulting mixture was stirred at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 8% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 26-7 (yellow solid, 3 g, 60% yield). MS (ESI, m/z) 767.4[ 2 ], M + H] ⁺ 。

Step 8

To a solution of 26-7 (3.00 g, 3.716 mmol, 1.00 eq) in methanol (40 ml) at 25 c under nitrogen with stirring was added acetic acid (1.17 g, 18.580 mmol, 5.00 eq) and palladium hydroxide on carbon (20% palladium content, 500 mg). The nitrogen gas is replaced by hydrogen gas by a replacement gas. The resulting mixture was stirred to react at 25 ℃ for 4 hours under a hydrogen atmosphere (1.5 atm). The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, insoluble matter was removed by filtration, the cake was washed with methanol (20 ml), and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 10% → 95% acetonitrile/water mobile phase (0.1% formic acid) over 20 minutes; detector UV220 nm; compound 26-8 was obtained (white solid, 2.0 mg, 79% yield). MS (ESI, m/z) 677.5[ 2 ] M + H ] ⁺ 。

Step 9

26-8 (500.0 mg, 0.702 mmol, 1.00 eq), 26-3 (336.66 mg, 0.912 mmol, 1.30 eq), and three were stirred at 25 deg.C under nitrogen protectionTo a solution of butylphosphine (298.91 mg, 1.404 mmol, 2.00 eq) in toluene (10 ml) was added azodicarbonamide (254.39 mg, 1.404 mmol, 2.00 eq). The resulting mixture was stirred at 60 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compounds 26-9 (white solid, 400 mg, 56% yield). MS (ESI, m/z) 1009.8[ m ] +H] ⁺ 。

Step 10

Tetrabutylammonium fluoride (124.33 mg, 0.452 mmol, 1.20 equivalents) was added to a solution of compound 26-9 (400.00 mg, 0.376 mmol, 1.00 equivalent) in tetrahydrofuran (4.0 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compounds 26-10 (colorless oil, 300 mg, yield 93%). MS (ESI, m/z) 853.6[ m ] +H ] ⁺ 。

Step 11

To a solution of compound 26-10 (300.00 mg, 0.334 mmol, 1.00 eq) in acetonitrile (6.0 ml) was added 2-iodoxybenzoic acid (147.71 mg, 0.501 mmol, 1.50 eq) with stirring at 25 ℃. The mixture was reacted at 60 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature. And concentrating the reaction solution under reduced pressure to obtain a crude product. Crude productThe product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 26-11 (white solid, 250 mg, 87% yield). MS (ESI, m/z) 851.5[ 2 ], [ M + H ]] ⁺ 。

Step 12

To a solution of compounds 26-11 (70.00 mg, 0.078 mmol, 1.00 eq) and 2-9a (39.22 mg, 0.063 mmol, 0.80 eq) in methanol (1.5 ml) was added acetic acid (4.94 mg, 0.078 mmol, 1.00 eq) and sodium cyanoborohydride (10.34 mg, 0.156 mmol, 2.00 eq) with stirring at zero degrees centigrade. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent to give an intermediate. To this intermediate was added dichloromethane (1.5 ml) and trifluoroacetic acid (0.5 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X150 mm, 5 μm; mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a phase b gradient of 85% → 95% in 7 minutes; detector UV 254/220 nm; yield 28a (white solid, 14.1 mg, 13% yield). MS (ESI, m/z) 1230.7/1232.7[ M + H ] ] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.51(s,2H),7.96(d,J＝1.6Hz,1H),7.75(d,J＝8.4Hz,1H),7.50–7.39(m,1H),7.29–7.16(m,4H),7.14–7.03(m,4H),6.82–6.76(m,4H),5.01–4.92(m,3H),4.59–4.52(m,3H),4.46–4.41(m,1H),4.24–4.20(m,1H),4.02–3.97(m,6H),3.82–3.74(m,5H),3.66–3.52(m,9H),3.28–3.19(m,7H),2.81(s,3H),2.74–2.67(m,2H),2.54(s,1H),2.49–2.34(m,3H),2.38–2.33(m,1H),2.22–2.19(m,2H),1.93–1.59(m,10H),1.44–1.34(m,3H),1.20–0.99(m,5H)。

Step 12'

To a solution of compounds 26-11 (50.00 mg, 0.056 mmol, 1.00 equiv.) and 2-9b (28.02 mg, 0.045 mmol, 0.80 equiv.) in methanol (1.0 ml) was added acetic acid (3.35 mg, 0.056 mmol, 1.00 equiv.) and sodium cyanoborohydride (7.38 mg, 0.112 mmol, 2.00 equiv.) with stirring at zero degrees centigrade. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent to give an intermediate. To this intermediate was added dichloromethane (1.0 ml) and trifluoroacetic acid (0.3 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18 OBD Column,19X150 mm, 5 μm; a mobile phase A: water (0.1% trifluoroacetic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; gradient: elution with a phase B gradient of 85% → 95% in 7 minutes; detector UV 254/220 nm; yield 28b (white solid, 5.8 mg, 8% yield). MS (ESI, m/z) 1230.7/1232.7[ m + H ] ] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.37(s,2H),7.88(d,J＝1.6Hz,1H),7.66(d,J＝8.4Hz,1H),7.34–7.29(m,1H),7.19–7.07(m,4H),7.05–6.94(m,4H),6.74–6.63(m,4H),4.92–4.82(m,3H),4.49–4.44(m,3H),4.38–4.32(m,1H),4.14–4.10(m,1H),3.94–3.89(m,6H),3.81–3.59(m,6H),3.58–3.51(m,8H),3.25–3.22(m,7H),2.77(s,3H),2.69–2.59(m,2H),2.50–2.44(m,3H),2.39–2.24(m,2H),2.17–2.10(m,2H),1.87–1.77(m,1H),1.72–1.57(m,7H),1.52–1.50(m,2H),1.39–1.29(m,3H),1.19–0.87(m,5H)。

Example 27

(2r, 3s,4r, 5s) -3- (3-chloro-2-fluorophenyl) -4- (4-chloro-2-fluorophenyl) -N- (4- ((15- ((S or R) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -12-methyl-3, 6, 9-trioxy-12-azapentadecyl) carbamoyl) -2-methoxyphenyl) -4-cyano-5-neopentylpyrrolidine-2-carboxamide 27a; (2R, 3S,4R, 5S) -3- (3-chloro-2-fluorophenyl) -4- (4-chloro-2-fluorophenyl) -N- (4- ((15- ((R or S) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) -4- (piperazin-1-yl) quinazolin-2-yl) oxy) -12-methyl-3, 6, 9-trioxy-12-azapentadecyl) carbamoyl) -2-methoxyphenyl) -4-cyano-5-neopentylpyrrolidine-2-carboxamide 27b.

Step 1

To a 50 ml round bottom flask were added, with stirring at 25 degrees celsius, 4- ((2r, 3s,4r, 5s) -3- (3-chloro-2-fluorophenyl) -4- (4-chloro-2-fluorophenyl) -4-cyano-5-neopentylpyrrolidine-2-carboxamide) -3-methoxybenzoic acid (400.0 mg, 0.6 mmol, 1.0 eq), dichloromethane (10.0 ml), 2- (7-azobenzotriazol) -N, N' -tetramethylurea hexafluorophosphate (296.1 mg, 0.7 mmol, 1.2 eq), 1-amino-3, 6, 9-trioxa-11-undecanol (150.5 mg, 0.7 mmol, 1.2 eq) and N, N-diisopropylethylamine (251.6 g, 1.9 mmol, 3.0 eq) in that order. The mixture was stirred at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the crude product was obtained by concentration under reduced pressure, purified by reverse phase flash chromatography (C18 column) eluting with 5% → 60% acetonitrile/water (0.1% ammonium bicarbonate) mobile phase over 25 minutes; detection of UV254 nm; compound 27-1 was obtained (white solid, 490.0 mg, 90% yield). MS (ESI, m/z) 791.2/793.2[ m + H ]] ⁺ ； ¹ HNMR(400MHz,DMSO-d ₆ )δ10.42(s,1H),8.52(t,J＝5.6Hz,1H),8.32(d,J＝8.4Hz,1H),7.78–7.71(m,1H),7.62–7.47(m,4H),7.44–7.32(m,3H),4.67–4.56(m,3H),4.43–4.34(m,1H),4.00–3.90(m,4H),3.67–3.48(m,11H),3.42–3.38(m,4H),1.68–1.62(m,1H),1.27(d,J＝13.9Hz,1H),0.98(s,9H)。

Step 2

Compound 27-1 (300 mg, 0.4 mmol, 1.0 eq), acetonitrile (10.0 ml) and 2-iodoxybenzoic acid (159.2 mg, 0.5 mmol, 1.5 eq) were added sequentially to a 50 ml reaction flask at 25 ℃ with stirring under nitrogen. The resulting mixture was stirred at 60 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is cooled to room temperature, and the reaction liquid is concentrated under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, yielding compound 27-2 (white solid, 290.0 mg, 92% yield). MS (ESI, m/z) 789.2/791.2[ 2 ], [ M + H ]] ⁺ 。

Step 3

To a solution of compound 2-9a (30.0 mg, 0.05 mmol, 1.0 eq) in methanol (5.0 ml) was added 27-2 (48.0 mg, 0.06 mmol, 1.3 eq), acetic acid (4.2 mg, 0.07 mmol, 1.5 eq) and sodium cyanoborohydride (8.8 mg, 0.14 mmol, 3.0 eq) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by reduced pressure concentration It is prepared by pulverizing the above materials. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 27-3a (white solid, 52.0 mg, 86% yield). MS (ESI, m/z) 1368.4/1370.4[ m ] +H] ⁺ 。

Step 4

To a solution of compound 27-3a (52.0 mg, 0.04 mmol, 1.0 eq) in dichloromethane (2.0 ml) was added trifluoroacetic acid (2.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by decompression and concentration. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: column xselectricity CSH Prep C18 OBD,19 × 250 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was performed with 20% → 50% B phase in 7 minutes, detector UV220/235 nm; compound 27a was obtained (white solid, 28.4 mg, 59% yield). MS (ESI, m/z) 1268.6/1270.6[ 2 ], [ M + H ]] ⁺ ； ¹ HNMR(400MHz,DMSO-d ₆ )δ10.41(s,1H),8.49–8.46(m,1H),8.32(d,J＝8.4Hz,1H),8.20(s,1H),7.95(d,J＝1.5Hz,1H),7.81(d,J＝8.3Hz,1H),7.76–7.70(m,1H),7.61–7.51(m,3H),7.50–7.47(m,1H),7.46–7.41(m,1H),7.39–7.33(m,3H),7.29(d,J＝2.4Hz,1H),7.24–7.18(m,2H),7.06(d,J＝2.3Hz,1H),4.62–4.55(m,2H),4.41–4.32(m,3H),3.98–3.93(m,1H),3.91(s,3H),3.86–3.79(m,4H),3.50–3.42(m,12H),3.41–3.36(m,2H),3.04–2.98(m,4H),2.49–2.46(m,2H),2.19(s,3H),1.92–1.83(m,2H),1.68–1.60(m,1H),1.34–1.20(m,2H),0.97(s,9H)。

Step 3'

Take a photograph at 25To a solution of compound 2-9b (40.0 mg, 0.06 mmol, 1.0 eq) in methanol (5.0 ml) was added 27-2 (64.0 mg, 0.08 mmol, 1.3 eq), acetic acid (5.6 mg, 0.09 mmol, 1.5 eq) and sodium cyanoborohydride (11.7 mg, 0.2 mmol, 3.0 eq) with stirring. The mixture was reacted at 25 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the reaction solution is concentrated under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 27-3b (white solid, 52.0 mg, 61% yield). MS (ESI, m/z) 1368.4/1370.4[ 2 ], [ M + H ] ] ⁺ 。

Step 4'

To a solution of compound 27-3b (52.0 mg, 0.04 mmol, 1 eq) in dichloromethane (4.0 ml) was added trifluoroacetic acid (2.0 ml) with stirring at 25 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the mixture is purified by preparative high performance liquid chromatography, wherein the purification conditions are as follows: column xselectricity CSH Prep C18 OBD,19 × 250 mm, 5 μm; a mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was performed with 20% → 50% B phase in 7 minutes, detector UV 220/235 nm; compound 27b was obtained (white solid, 19.1 mg, 40% yield). MS (ESI, m/z) 1268.6/1270.6[ m ] +H] ⁺ ； ¹ HNMR(400MHz,DMSO-d ₆ )δ10.41(s,1H),8.52–8.44(m,1H),8.32(d,J＝8.4Hz,1H),8.19(s,1H),7.96(d,J＝1.6Hz,1H),7.84–7.69(m,2H),7.62–7.32(m,8H),7.29(d,J＝2.4Hz,1H),7.26–7.17(m,2H),7.07(d,J＝2.3Hz,1H),4.62–4.55(m,2H),4.42–4.31(m,3H),4.01–3.94(m,1H),3.91(s,3H),3.87–3.80(m,4H),3.52–3.44(m,12H),3.43–3.36(m,2H),3.07–3.00(m,4H),2.49–2.46(m,2H),2.20(s,3H),1.93–1.81(m,2H),1.70–1.58(m,1H),1.32–1.22(m,2H),0.97(s,9H)。

Example 28

(2s, 4r) -1- ((S) -2- (tert-butyl) -17- (4- ((S or R) -6-chloro-2- (3- (dimethylamino) azetidin-1-yl) -8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-4-yl) piperazin-1-yl) -4-oxo-6, 9,12, 15-tetraoxo-3-azaheptanoyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 28a; (2S, 4R) -1- ((S) -2- (tert-butyl) -17- (4- ((R or S) -6-chloro-2- (3- (dimethylamino) azetidin-1-yl) -8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-4-yl) piperazin-1-yl) -4-oxo-6, 9,12, 15-tetraoxo-3-azaheptanoyl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 28b.

Step 1

To a solution of the compound tert-butyl acetate-tetraethylene glycol (1.0 g, 3.2 mmol, 1.0 eq.) in dichloromethane (10.0 ml) was added trifluoroacetic acid (3.0 ml) with stirring at 25 ℃. The mixture was stirred at 25 ℃ for 4 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration under reduced pressure gave crude 28-1 (bright yellow oil, 820.0 mg). The crude product was used directly in the next reaction. MS (ESI, m/z) 253.1[ 2 ], [ M ] +H] ⁺ 。

Step 2

To a solution of compound 28-1 (818.5 mg, 1.0 eq) in N, N-dimethylformamide (8.0 ml) was added 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexakis (7-azobenzotriazol) with stirring at 25 deg.CFluorophosphate (1.4 g, 3.6 mmol, 1.1 eq). The mixture was reacted at 25 ℃ for 30 minutes. Then N, N-diisopropylethylamine (2.1 g, 16.3 mmol, 5.0 equivalents) and (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) were added to the reaction mixture]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (1.3 g, 2.6 mmol, 0.8 eq). The mixture was reacted at 25 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 25 minutes; detector UV254/220 nm; compound 28-2 was obtained (bright yellow solid, 948.5 mg, 40% yield). MS (ESI, m/z) 679.4[ m + H ], [ M ], [ phi ] ] ⁺ 。

Step 3

Compound 28-2 (785.0 mg, 1.2 mmol, 1.0 eq), acetonitrile (8.0 ml) and 2-iodoxybenzoic acid (356.4 mg, 1.3 mmol, 1.5 eq) were added sequentially to a 100 ml reaction flask at 25 degrees c with stirring under nitrogen. The resulting mixture was stirred at 60 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is cooled to room temperature, and a crude product is obtained by decompression and concentration. The crude product was purified by silica gel column chromatography eluting with a methanol/dichloromethane mobile phase gradient of 0% → 10% and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 28-3 (white solid, 615.3 mg, 79% yield). MS (ESI, m/z) 677.2[ m + H ], [ M ], [ phi ]] ⁺ 。

Step 4

Under the condition of stirring at 25 ℃ under the protection of nitrogen gas, the mixture is transformed into the liquidTo a solution of compounds 1-7 (500.0 mg, 1.0 mmol, 1.0 eq) in N-methylpyrrolidinone (5.0 ml) was added 3- (dimethylamino) azetidine dihydrochloride (135.6 mg, 1.3 mmol, 1.3 eq), N-diisopropylethylamine (722.4 mg, 5.6 mmol, 4.0 eq). The mixture was stirred at 60 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was cooled to room temperature. The reaction solution was directly purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water (0.1% ammonium bicarbonate) mobile phase over 25 minutes; detector UV254/220 nm; compound 28-4 was obtained (white solid, 900 mg, 29% yield). MS (ESI, m/z) 543.1/545.0/547.0M + H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.62–7.62(m,1H),4.36–4.16(m,2H),4.15–3.96(m,2H),3.75–3.50(m,8H),3.28–3.14(m,1H),2.24(s,6H),1.49(s,9H)。

Step 5

To a solution of compound 28-4 (900.0 mg, 1.7 mmol, 1.0 eq) in tetrahydrofuran/water (10/1, 11.0 ml) was added 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (535.0 mg, 2.0 mmol, 1.2 eq), potassium carbonate (684.7 mg, 5.0 mmol, 3.0 eq), and chlorine (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1, 1' -biphenyl) [2- (2 '-amino-1, 1' -biphenyl ] at 25 deg.C with stirring under nitrogen protection]Palladium (II) (129.7 mg, 0.2 mmol, 0.1 eq). The reaction solution was stirred at 60 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was cooled to room temperature. The reaction solution was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 28-5 (mixture of two stereoisomers, white solid, 600.0 mg, yield 60%) was obtained. MS (ESI, m/z) 607.2/609.2[ M + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.70–7.62(m,2H),7.60–7.55(m,1H),7.38–7.27(m,2H),7.23–7.20(m,1H),7.19–7.08(m,1H),4.20–4.40(m,2H),4.16–3.98(m,2H),3.82–3.50(m,8H),3.22–3.08(m,1H),2.18(s,6H),1.530(s,9H)。

Step 6

Chiral resolution of the compound 28-5 (600.0 mg) obtained in step 5 by preparative chiral high performance liquid chromatography: chiral column CHIRALPAK IC,2 × 25 cm, 5 microns; mobile phase A: n-hexane/methyl tert-butyl ether =1/1 (0.5% 2 mol/l ammonia methanol), mobile phase B: ethanol; flow rate: 20 ml/min; elution was performed with 10% phase B in 11.4 minutes, detector UV225/254 nm. Two products were obtained, a shorter retention time (6.8 minutes) product being compound 28-5a (white solid, 225.0 mg, 37% recovery), compound 28-5a: MS (ESI, m/z) 607.2/609.2[ M + H ] ] ⁺ (ii) a The product at longer retention time (9 min) was compound 28-5b (white solid, 230.0 mg, 38% recovery), compound 28-5b: MS (ESI, m/z) 607.2/609.2[ M + H ]] ⁺ 。

Step 7

To a solution of compound 28-5a (225.0 mg, 0.4 mmol, 1.0 eq) in dichloromethane (4.0 ml) was added trifluoroacetic acid (1.0 ml) with stirring at zero degrees centigrade. The mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Purifying the crude product by using a high performance liquid chromatography, wherein the purification conditions are as follows: chromatographic column Gemini-NX C18 AXAI Packed,21.2x150 mm, 5 microns, mobile phase a: water (10 mol/l ammonium bicarbonate), mobile phase B acetonitrile; flow rate: 25 ml/min; elution with 10% mobile phase B for 2 minutes, followed by 10% → 26% streamMobile phase B eluted for 2.5 minutes and finally for 2 minutes with 26% → 55% mobile phase B. Detector UV220 nm; product 28-6a was obtained (white solid, 150.0 mg, 80% yield). MS (ESI, m/z): 507.2/509.1[ M ] +H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ10.00(s,1H),7.83–7.72(m,2H),7.49–7.38(m,1H),7.30–7.18(m,3H),7.04(d,J＝2.4Hz,1H),4.16–4.05(m,2H),3.92–3.81(m,2H),3.70–3.60(m,4H),3.18–3.06(m,1H),2.96–2.82(m,4H),2.11(s,6H)。

Step 7'

To a solution of compound 28-5b (230.0 mg, 0.4 mmol, 1.0 eq) in dichloromethane (4.0 ml) was added trifluoroacetic acid (1.0 ml) with stirring at zero degrees centigrade. The mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. Purifying the crude product by high performance liquid chromatography, wherein the purification conditions are as follows: chromatographic column Gemini-NX C18 AXAI Packed, 21.2xc150 mm, 5 μm, mobile phase a: water (10 mol/l ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; elute with 10% mobile phase B for 2 minutes, then elute with 10% → 26% mobile phase B for 2.5 minutes, and finally elute with 26% → 55% mobile phase B for 2 minutes. Detector UV220 nm; product 28-6b was obtained (white solid, 145.0 mg, 76% yield). MS (ESI, m/z): 507.2/509.1[ M ] +H ] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ9.99(s,1H),7.84–7.72(m,2H),7.49–7.38(m,1H),7.30–7.17(m,3H),7.04(d,J＝2.4Hz,1H),4.17–4.04(m,2H),3.93–3.81(m,2H),3.76–3.63(m,4H),3.18–3.07(m,1H),3.03–2.89(m,4H),2.12(s,6H)。

Step 8

Under the stirring condition of 25 ℃, the mixture is converted into the alcoholTo a solution of compound 28-6a (40.0 mg, 0.08 mmol, 1.0 eq) in methanol (2.0 ml) was added sodium cyanoborohydride (7.8 mg, 0.1 mmol, 1.5 eq) and acetic acid (7.5 mg, 0.1 mmol, 1.5 eq). Compound 28-3 (64.8 mg, 0.1 mmol, 1.2 eq) was then dissolved in methanol (2.0 ml) and slowly added dropwise to the reaction. The mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Purifying the crude product by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18OBD Column,9X250 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution with mobile phase B32% → 61% for 7 minutes; detector UV254/220 nm; product 28a was obtained (white solid, 31.0 mg, 34% yield). MS (ESI, m/z) 1167.4/1169.4[ M + H ]] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.90–8.86(m,1H),7.82–7.72(m,2H),7.48–7.34(m,5H),7.31–7.13(m,3H),7.04(d,J＝2.4Hz,1H),5.07–4.97(m,1H),4.70(s,1H),4.63–4.53(m,1H),4.48–4.41(m,1H),4.32–4.22(m,2H),4.09–3.99(m,4H),3.89–3.80(m,5H),3.79–3.63(m,15H),3.30–3.20(m,1H),2.86–2.65(m,6H),2.48(s,3H),2.26(s,6H),2.24–2.16(m,1H),2.10–1.90(m,1H),1.56–1.48(m,3H),1.11–1.01(m,9H)。

Step 8'

To a solution of compound 28-6b (50.0 mg, 0.1 mmol, 1.0 eq) in methanol (2.0 ml) was added sodium cyanoborohydride (9.3 mg, 0.1 mmol, 1.5 eq) and acetic acid (8.9 mg, 0.2 mmol, 1.5 eq) with stirring at 25 ℃. Compound 28-3 (80.1 mg, 0.1 mmol, 1.2 eq) was then dissolved in methanol (2.0 ml) and slowly added dropwise to the reaction. The mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Crude product tong Purifying by preparative high performance liquid chromatography under the following conditions: XSelect CSH Prep C18OBD Column,19X250 mm, 5 μm; a mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution with mobile phase B32% → 61% for 7 minutes; detector UV254/220 nm; product 28b was obtained (white solid, 31.0 mg, 28% yield). MS (ESI, m/z) 1167.4/1169.4[ 2 ] M + H] ⁺ ； ¹ H NMR(300MHz,CD ₃ OD)δ8.90–8.86(m,1H),7.82–7.72(m,2H),7.48–7.34(m,5H),7.31–7.14(m,3H),7.04(d,J＝2.4Hz,1H),5.08–4.96(m,1H),4.70(s,1H),4.63–4.53(m,1H),4.48–4.41(m,1H),4.32–4.22(m,2H),4.09–3.99(m,4H),3.90–3.80(m,5H),3.79–3.62(m,15H),3.30–3.20(m,1H),2.86–2.65(m,6H),2.48(s,3H),2.26(s,6H),2.24–2.16(m,1H),2.10–1.90(m,1H),1.56–1.48(m,3H),1.11–1.01(m,9H)。

Example 29

(2S, 4R) -1- ((S) -2- (2- (2- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) ethoxy) acetamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide dihydrochloride 29

Compound 29 was synthesized according to example 25. Compound 29 (light yellow solid). MS (ESI, m/z) 1048.5/1050.5[ M + H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ10.86–9.78(m,3H),9.09(s,1H),8.70–8.61(m,1H),8.01(d,J＝1.7Hz,1H),7.81(d,J＝8.4Hz,1H),7.73(d,J＝9.3Hz,1H),7.48–7.36(m,5H),7.32(d,J＝2.4Hz,1H),7.27–7.17(m,2H),7.12(d,J＝2.4Hz,1H),4.86–4.66(m,2H),4.59–4.32(m,6H),4.29–3.82(m,10H),3.80–3.56(m,4H),3.49–3.38(m,1H),3.31–3.20(m,1H),2.45(s,3H),2.31–2.16(m,1H),2.11–1.81(m,9H),0.92(s,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-121.87。

Example 30

(2S, 4R) -1- ((S) -2- (3- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) propanamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide dihydrochloride 30

Compound 30 was synthesized according to example 25. Compound 30 (light yellow solid). MS (ESI, m/z) 1018.6/1020.5[ m + H ]] ⁺ 。 ¹ H NMR(300MHz,DMSO-d ₆ )δ10.74(s,1H),9.99(s,1H),9.74(s,1H),9.05(s,1H),8.68–8.58(m,1H),8.27(d,J＝9.2Hz,1H),8.01(d,J＝1.6Hz,1H),7.82(d,J＝8.3Hz,1H),7.49–7.28(m,6H),7.27–7.15(m,2H),7.11(d,J＝2.4Hz,1H),4.88–4.64(m,3H),4.60–4.50(m,4H),4.27–4.23(m,1H),4.21–4.09(m,3H),4.07–3.88(m,3H),3.76–3.47(m,4H),3.44–3.28(m,1H),3.24–3.09(m,1H),2.95–2.73(m,2H),2.45(s,3H),2.34–2.15(m,1H),2.12–1.79(m,9H),0.91(s,9H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-121.79。

Example 31

(2S, 4R) -1- ((S) -2- (4- ((S) -2- ((((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) butanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide dihydrochloride 31

Compound 31 was synthesized according to example 25. Compound 31 (off-white solid). MS (ESI, m/z) 1032.6/1034.6[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ10.71(s,1H),10.09–9.99(m,1H),9.84–9.69(m,1H),9.12(s,1H),8.71–8.61(m,1H),8.16–8.05(m,2H),7.88(d,J＝8.3Hz,1H),7.55–7.42(m,5H),7.38(d,J＝2.4Hz,1H),7.34–7.24(m,2H),7.17(d,J＝2.4Hz,1H),4.90–4.72(m,2H),4.68–4.36(m,6H),4.32(d,J＝5.6Hz,1H),4.27–4.21(m,2H),4.09–3.95(m,3H),3.77–3.61(m,3H),3.55–3.40(m,1H),3.28–3.08(m,2H),2.52(s,3H),2.46–2.26(m,3H),2.01(d,J＝11.3Hz,11H),0.98(s,9H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-121.80。

Example 32

(2S, 4R) -1- ((2S) -2- (4- (4- ((2S) -2- (((7S or 7R) -4- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) butoxy) butyramido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide dihydrochloride 32

Compound 32 was synthesized according to example 25. Compound 32 (light yellow solid). MS (ESI, m/z): 1104.5/1106.5[ m ] +H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.47–10.34(m,1H),9.95–9.82(m,1H),9.66–9.59(m,1H),9.02(s,1H),8.64–8.54(m,1H),8.01(d,J＝1.6Hz,1H),7.94–7.76(m,2H),7.49–7.35(m,5H),7.31(d,J＝2.4Hz,1H),7.26–7.14(m,2H),7.09(d,J＝2.4Hz,1H),4.80–4.65(m,2H),4.59–4.49(m,3H),4.47–4.38(m,2H),4.37–4.31(m,1H),4.26–4.14(m,3H),4.00–3.85(m,4H),3.52–3.38(m,3H),3.34–3.26(m,4H),3.17–3.06(m,2H),2.44(s,3H),2.29–2.13(m,3H),2.08–1.86(m,9H),1.82–1.63(m,4H),1.57–1.46(m,2H),0.92(s,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-121.88。

Example 33

(2S, 4R) -1- ((S) -2- (7- ((S) -2- ((((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) heptanamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide dihydrochloride 33

Compound 33 was synthesized according to example 25. Compound 33 (light yellow solid). MS (ESI, m/z) 1174.6/1176.6[ 2 [ M ] +H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ9.01(s,1H),8.00(s,1H),7.80(d,J＝8.3Hz,1H),7.49–7.33(m,5H),7.31(d,J＝2.4Hz,1H),7.27–7.14(m,2H),7.06(d,J＝2.4Hz,1H),4.75–4.62(m,2H),4.60–4.47(m,3H),4.45–4.37(m,2H),4.37–4.31(m,1H),4.26–4.15(m,3H),3.93–3.87(m,3H),3.66–3.62(m,2H),3.45–3.34(m,1H),3.21–3.00(m,2H),2.42(s,3H),2.31–2.16(m,2H),2.14–1.79(m,10H),1.71–1.59(m,2H),1.51–1.36(m,2H),1.35–1.14(m,5H),0.89(s,9H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-121.87。

Example 34

(2s, 4r) -1- ((2S) -2- (2- (5- (2- ((2S) -2- (((7S or 7R) -4- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) ethyl) tetrahydrofuran-2-yl) acetamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide diformate 34; (2S, 4R) -1- ((2S) -2- (2- ((2S or R,5S or R) -5- (2- ((2S) -2- (((7S or 7R) -4- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) ethyl) tetrahydrofuran-2-yl) acetamide-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide formate 34aa; (2s, 4r) -1- ((2S) -2- (2- ((2S or R,5R or S) -5- (2- ((2S) -2- (((7S or 7R) -4- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) ethyl) tetrahydrofuran-2-yl) acetamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide formate 34ab; (2s, 4r) -1- ((2S) -2- (2- ((2R or S,5S or R) -5- (2- ((2S) -2- (((7S or 7R) -4- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) ethyl) tetrahydrofuran-2-yl) acetamide) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide diformate 34ba; (2S, 4R) -1- ((2S) -2- (2- ((2R or S,5R or S) -5- (2- ((2S) -2- (((7S or 7R) -4- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) ethyl) tetrahydrofuran-2-yl) acetamide-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide formate bb34

Step 1:

to a solution of ethyl 2-iodoacetate (20.0 g, 88.8 mmol, 1.0 equiv.), furan (114.5 g, 1598.1 mmol, 18.0 equiv.) and iron sulfate heptahydrate (13.0 g, 44.4 mmol, 0.5 equiv.) in dimethyl sulfoxide (200 ml) was added hydrogen peroxide (30%, 22.2 g, 195.3 mmol, 2.2 equiv.) with zero degrees centigrade stirring. The mixture was allowed to react for 4 hours at 20 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, water (1200 ml) was added to the reaction solution, followed by extraction with ethyl acetate (800 ml × 3), and the organic phases were combined; the organic phase was washed with saturated brine (800 ml x 3), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 30% methyl tert-butyl ether/petroleum ether and the resulting fraction was rotary distilled under reduced pressure to remove the solvent and give compound 34-1 (colorless oil, 10 g, 69% yield). MS (ESI, m/z) 155.0[ m + H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.40–7.32(m,1H),6.35–6.32(m,1H),6.24–6.21(m,1H),4.23–4.15(m,2H),3.68(d,J＝2.1Hz,2H),1.31–1.25(m,3H)。

And 2, step:

to a solution of compound 34-1 (10.0 g, 61.6 mmol, 1.0 eq) in anhydrous tetrahydrofuran (100 ml) was slowly added a solution of lithium aluminum hydride in tetrahydrofuran (2.5 mol/l, 49 ml, 123.2 mmol, 2.0 eq) dropwise with stirring at zero degrees centigrade under nitrogen protection. The mixture was reacted at 0 ℃ for 1.5 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, water (4.9 g), a 10% aqueous solution of sodium hydroxide (4.9 g), and water (14.7 g) were gradually added dropwise to the reaction mixture at 0 ℃. The insoluble material was removed by filtration, the filter cake was washed with tetrahydrofuran (60 ml x 3), and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 30% ethyl acetate/petroleum ether and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 34-2 (yellow oil, 4.86 g, 66% yield). ¹ H NMR(400MHz,CDCl ₃ )δ7.39–7.29(m,1H),6.35–6.30(m,1H),6.14–6.06(m,1H),3.88(t,J＝6.2Hz,2H),2.90(t,J＝6.2Hz,2H)。

And step 3:

to a solution of compound 34-2 (4.45 g, 37.7 mmol, 1.0 eq), ethyl 2-iodoacetate (8.5 g, 37.7 mmol, 1.0 eq) and iron sulfate heptahydrate (5.5 g, 18.9 mmol, 0.5 eq) in dimethyl sulfoxide (80 ml) was slowly added hydrogen peroxide (30%, 9.4 g, 83.0 mmol, 2.2 eq) with stirring at zero degrees centigrade. The mixture was reacted at 20 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction is carried outWater (500 ml) was added to the reaction solution, followed by extraction with ethyl acetate (400 ml × 3), and the organic phases were combined; the organic phase was washed with saturated brine (800 ml x 3), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 50% ethyl acetate/petroleum ether and the resulting fraction was rotary distilled under reduced pressure to remove the solvent and afford compound 34-3 (yellow oil, 3.8 g, 45% yield). MS (ESI, m/z) 199.0[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ6.13(d,J＝3.1Hz,1H),6.05(d,J＝3.1Hz,1H),4.22–4.15(m,2H),3.87–3.81(m,2H),3.64(s,2H),2.90–2.80(m,2H),1.84(s,1H),1.30–1.23(m,3H)。

And 4, step 4:

rhodium/alumina (5%, 1.9 g, 0.9 mmol, 0.05 eq.) was added to a solution of compound 34-3 (3.8 g, 18.2 mmol, 1.0 eq.) in ethanol (50 ml) at 25 deg.c with stirring under nitrogen. The nitrogen was then replaced by hydrogen. The mixture was reacted at 25 ℃ for 40 hours in a hydrogen atmosphere of 5 atmospheres, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, insoluble matter was removed by filtration, the filter cake was washed with ethanol (20 × 3 ml), and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 70% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 34-4 (yellow oil, 2.05 g, 52% yield). MS (ESI, m/z) 203.0[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.33–4.21(m,1H),4.20–4.11(m,2H),4.11–4.02(m,1H),3.81–3.75(m,2H),2.66–2.55(m,1H),2.54–2.43(m,2H),2.16–1.98(m,2H),1.90–1.73(m,2H),1.72–1.55(m,2H),1.27(t,J＝7.1Hz,3H)。

And 5:

to a solution of compound 34-4 (2.0 g, 9.4 mmol, 1.0 eq), 4-dimethylaminopyridine (0.12 g, 0.94 mmol, 0.1 eq) and triethylamine (2.0 g, 18.8 mmol, 2.0 eq) in dichloromethane (20 ml) was slowly added p-toluenesulfonyl chloride (2.1 g, 10.3 mmol, 1.1 eq) with zero degrees centigrade stirring. The reaction solution was reacted at 25 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, adding water (20 ml) into the system to quench the reaction, extracting the water phase with dichloromethane (20 ml × 3), and combining the organic phases; the organic phase is dried by anhydrous sodium sulfate, the drying agent is removed by filtration, and the filtrate is concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a gradient of 0% → 50% ethyl acetate/petroleum ether and the fractions were freed from the solvent by rotary evaporation under reduced pressure to give compound 34-5 (colorless oil, 3.2 g, 90% yield). MS (ESI, m/z) 357.1[ 2 ] M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.83–7.76(m,2H),7.40–7.32(m,2H),4.21–4.07(m,5H),3.94–3.80(m,1H),2.58–2.34(m,5H),2.12–1.80(m,4H),1.63–1.46(m,2H),1.30–1.22(m,3H)。

Step 6:

compound 34-5 (3.0 g, 8.0 mmol, 1.0 eq) was dissolved in 30 ml of acetonitrile with stirring at 25 ℃, followed by addition of L-prolinol (1.02 g, 9.6 mmol, 1.2 eq) and potassium carbonate (2.33 g, 16 mmol, 2.0 eq). The mixture was reacted at 60 ℃ for 4 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, insoluble material was removed by filtration, the filter cake was washed with acetonitrile (15 × 2 ml), and the filtrate was concentrated under reduced pressure to give a crude product. Purifying the crude product by silica gel column chromatography, eluting mobile phase with 0% → 10% ammonia methanol/dichloromethane gradient, removing solvent from the obtained fraction by rotary evaporation under reduced pressure to obtain To compound 34-6 (yellow oil, 2.27 g, 94% yield). MS (ESI, m/z) 286.1[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ4.29–4.19(m,1H),4.19–4.08(m,2H),4.02–3.85(m,1H),3.68–3.59(m,1H),3.45–3.33(m,1H),3.27–3.14(m,1H),2.95–2.80(m,1H),2.66–2.56(m,2H),2.49–2.21(m,3H),2.15–1.97(m,2H),1.91–1.49(m,9H),1.29–1.24(m,3H)。

And 7:

compound 34-6 (1.2 g, 4.0 mmol, 1.0 eq) was dissolved in a tetrahydrofuran/methanol/water mixture (1/1/1, 12 ml) with stirring at 25 ℃, followed by addition of lithium hydroxide (0.2 g, 7.9 mmol, 2.0 eq). The mixture was allowed to react at 25 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove the solvent, and then 12 ml of water was added to dilute the mixture. Dilute hydrochloric acid (1 mol/l, 8 ml) was added dropwise at 0 ℃ to adjust the pH to 5, followed by concentration under reduced pressure to give crude compound 34-7 (1.4 g). The crude product was used directly in the next reaction without purification. MS (ESI, m/z) 258.1[ 2 ] M + H] ⁺ 。

And 8:

to a solution of compound 16-3a (1.0 g, 1.5 mmol, 1.0 equiv) and compound 34-7 (0.71 g, 1.8 mmol, 1.2 equiv, 67% pure) in anhydrous tetrahydrofuran (10 ml) was added a solution of potassium tert-butoxide in tetrahydrofuran (1 mol/l, 3.9 ml, 3.9 mmol, 2.5 equiv) under zero degrees centigrade nitrogen blanket. The mixture was reacted at zero degrees centigrade for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the crude product was purified by reverse phase chromatography (C18 column) using a mobile phase of 5% → 95% methanol/water over a period of 25 minutes Eluting with 0.1% ammonia water; detector, UV254/220 nm; compound 34-8 was obtained (yellow solid, 700 mg, 51% yield). MS (ESI, m/z) 834.4/836.4[ M + H ]] ⁺ 。

And step 9:

to a solution of compounds 34-8 (700 mg, 0.84 mmol, 1.0 eq) in N, N-dimethylformamide (8 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (383 mg, 1.01 mmol, 1.2 eq) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 15 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group was added to the reaction mixture]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (472 mg, 1.01 mmol, 1.2 eq) and N, N-diisopropylethylamine (433 mg, 3.36 mmol, 4 eq). The reaction solution was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, water (60 ml) was added to the reaction flask, followed by extraction with ethyl acetate (60 ml × 3). The organic phases were combined, washed with saturated brine (30 ml x 3), dried over anhydrous sodium sulfate and filtered to remove the drying agent; concentrating the filtrate under reduced pressure to obtain a crude product. The mixture was purified by reverse phase chromatography (C18 column) eluting with 40% → 75% methanol/water mobile phase (0.1% ammonium bicarbonate) over 25 minutes; detector, UV254/220 nm; compound 34-9 was obtained (yellow solid, 750 mg, 72% yield). MS (ESI, m/z) 1246.5/1248.5[ 2 ], [ M ] +H ] ⁺ 。

Step 10:

to a solution of compound 34-9 (50 mg, 0.038 mmol, 1.0 eq) in methanol (2 ml) was added dropwise a solution of hydrochloric acid in 1, 4-dioxane (4) with stirring at 0 deg.CMol/l, 2 ml). The mixture was allowed to react for 2 hours at 25 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is decompressed and concentrated to obtain a crude product. The crude product was dissolved in methanol (2 ml) and methanolic ammonia (7 mol/l, 2 ml) was added with stirring at 0 ℃. After the mixture was stirred at 25 ℃ for 20 minutes, it was then concentrated under reduced pressure. The crude product was dissolved in methanol (2 ml) and formic acid (2 ml) was added with stirring at 0 ℃. After the mixture was stirred at 25 ℃ for 10 minutes, it was then concentrated under reduced pressure to give a crude product. Purifying the obtained crude product by high performance liquid chromatography with a chromatographic column Sunfire prep C18 column,30x150 mm, 5 microns; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution was performed with 32% → 51% B phase over 7 minutes, detector UV 220/254 nm. Compound 34 was obtained (white solid, 14 mg, 30% yield). MS (ESI, m/z) 1102.1/1104.1[ 2 ], [ M + H ] ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.97–8.94(m,1H),8.63–8.57(m,1H),8.29(s,2H),7.98–7.75(m,3H),7.48–7.34(m,5H),7.30(d,J＝2.4Hz,1H),7.27–7.17(m,2H),7.07(d,J＝2.4Hz,1H),4.58–3.96(m,11H),3.81–3.63(m,7H),3.09–2.81(m,3H),2.48–2.34(m,5H),2.28–2.16(m,2H),2.08–1.99(m,1H),1.96–1.82(m,4H),1.81–1.58(m,9H),1.53–1.38(m,2H),0.96–0.84(m,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-122.27。

Step 11

Chiral resolution of the compound 34-9 (700.0 mg) obtained in step 9 by preparative supercritical liquid chromatography: chiral column CHIRALPAK IA,3 × 25 cm, 5 microns; mobile phase A: supercritical carbon dioxide, mobile phase B: methanol/dichloromethane =2/1 (0.1% 2 moles/liter ammonia methanol); flow rate: 70 ml/min; elution was performed with 40% phase B over 12 minutes with detector UV220/235 nm to give two products. The product with shorter retention time (6.68 min) was compound 34-9a (C.) (Mixture of two stereoisomers, yellow oil, 340 mg, 48% recovery), compound 34-9a: MS (ESI, m/z) 1246.5/1248.5[ 2 ], [ M ] +H] ⁺ (ii) a The product of longer retention time (10.32 min) was compound 34-9b (mixture of two stereoisomers, yellow oil, 280 mg, 40% recovery), compound 34-9b: MS (ESI, m/z) 1246.5/1248.5[ m + H ], [] ⁺ 。

Step 12

Chiral resolution of compound 34-9a (340 mg) from step 11 by preparative supercritical liquid chromatography: chiral column CHIRALPAK IA,3 × 25 cm, 5 microns; mobile phase A: supercritical carbon dioxide, mobile phase B: methanol (0.5% by 2 mol/l ammonia methanol); flow rate: 70 ml/min; elution was performed with 30% phase B over 10 minutes, detector UV 222 nm, yielding two products. The product of shorter retention time (6.12 min) was compound 34-9aa (yellow oil, 12 mg, 3% recovery), compound 34-9aa: MS (ESI, m/z) 1246.5/1248.5[ m + H ], [ ] ⁺ (ii) a The product at longer retention time (6.78 min) was compound 34-9ab (yellow oil, 290 mg, 85% recovery), compound 34-9ab: MS (ESI, m/z) 1246.5/1248.5[ 2 ], [ M ] +H] ⁺ 。

Step 13

Chiral resolution of compound 34-9b (280 mg) from step 11 by preparative supercritical liquid chromatography: chiral column CHIRALPAK IA,3 × 25 cm, 5 micron; a mobile phase A: supercritical carbon dioxide, mobile phase B: methanol (0.5% 2 mol/l ammonia methanol); flow rate: 70 ml/min; elution was performed with 30% phase B over 10 minutes, detector UV 222 nm, yielding two products. The product of shorter retention time (4.8 min) was compound 34-9ba (yellow oil, 160 mg, 57% recovery), compound 34-9ba: m is a group ofS(ESI,m/z):1246.5/1248.5[M+H] ⁺ (ii) a The product at longer retention time (6.03 min) was compound 34-9bb (yellow oil, 50 mg, 17% recovery), compound 34-9bb: MS (ESI, m/z) 1246.5/1248.5[ m + H ], [] ⁺ 。

Step 14

Using the procedure in step 10, starting from compounds 34-9aa, compound 34aa was obtained (white solid, 6 mg, 53% yield). MS (ESI, m/z) 1102.1/1104.1[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.84(s,1H),8.72–8.40(m,1H),7.98–7.92(m,1H),7.74(d,1H),7.47–7.32(m,5H),7.26(d,J＝2.4Hz,1H),7.24–7.15(m,2H),7.04–7.01(m,1H),4.68–4.43(m,8H),4.41–4.34(m,1H),4.30–4.23(m,1H),4.12–4.01(m,1H),3.88–3.80(m,1H),3.80–3.61(m,5H),3.60–3.35(m,4H),3.07–2.96(m,1H),2.86–2.73(m,1H),2.48–2.32(m,5H),2.22–2.13(m,2H),2.10–1.99(m,3H),1.99–1.75(m,9H),1.66–1.52(m,2H),0.98(s,9H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-122.92。

Step 15

Using the procedure in step 10, starting from compound 34-9ab, 34ab was obtained (white solid, 41.2 mg, 49% yield). MS (ESI, m/z) 1102.1/1104.1[ 2 ], [ M + H ] ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.96(s,1H),8.32–8.19(m,1H),7.97–7.89(m,1H),7.80(d,J＝8.7Hz,2H),7.48–7.34(m,5H),7.29(d,J＝2.4Hz,1H),7.26–7.17(m,2H),7.06(d,J＝2.3Hz,1H),4.55–4.48(m,1H),4.48–4.29(m,6H),4.26–4.19(m,1H),4.13–3.95(m,2H),3.81–3.69(m,3H),3.68–3.57(m,5H),3.08–2.89(m,2H),2.88–2.79(m,1H),2.49–2.32(m,5H),2.28–2.17(m,2H),2.09–1.95(m,1H),1.95–1.82(m,4H),1.79–1.60(m,9H),1.50–1.42(m,2H),0.89(s,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-122.21,-122.25。

Step 16

Using the procedure in step 10, compound 34-9ba was used as the starting material to give 34ba (white solid, 37.2 mg, 50% yield). MS (ESI, m/z) 1102.1/1104.1[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.97(s,1H),8.62–8.56(m,1H),8.24–8.19(m,2H),7.95–7.87(m,2H),7.80(d,J＝8.3Hz,1H),7.49–7.35(m,5H),7.28(d,J＝2.4Hz,1H),7.24–7.19(m,2H),7.06(d,J＝2.4Hz,1H),4.57–4.30(m,8H),4.25–4.18(m,1H),4.11–4.05(m,1H),4.01–3.95(m,1H),3.74–3.70(m,2H),3.59–3.55(m,5H),3.06–3.01(m,1H),2.90–2.78(m,2H),2.43–2.37(m,6H),2.23–2.14(m,1H),2.07–1.98(m,1H),1.93–1.78(m,4H),1.77–1.42(m,11H),0.91(s,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-122.29。

Step 17

Using the procedure in step 10, starting from compounds 34-9bb, gave 34bb (white solid, 18 mg, 38% yield). MS (ESI, m/z): 1102.1/1104.1[ m ] +H] ⁺ ； ¹ H NMR(400MHz,CD ₃ OD)δ8.85(s,1H),8.53(s,1H),7.95(d,J＝1.7Hz,1H),7.74(d,J＝8.3Hz,1H),7.48–7.33(m,5H),7.26(d,J＝2.4Hz,1H),7.22–7.14(m,2H),7.07–6.98(m,1H),4.71–4.59(m,3H),4.57–4.43(m,5H),4.39–4.32(m,1H),4.27–4.18(m,1H),4.16–4.07(m,1H),3.88–3.46(m,9H),3.07–2.87(m,2H),2.49–2.33(m,5H),2.28–2.14(m,2H),2.13–1.94(m,6H),1.94–1.75(m,6H),1.65–1.46(m,2H),1.36–1.22(m,1H),1.07–0.93(m,8H),0.92–0.82(m,1H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-122.84。

Example 35

(2S, 4R) -1- ((S) -2- (2- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) acetamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide dihydrochloride 35

Step 1:

to a solution of L-prolinol (3.00 g, 29.6 mmol, 1.00 eq) and allyl chloride (2.27 g, 29.6 mmol, 1.00 eq) in acetonitrile (30 ml) was added potassium carbonate (8.20 g, 59.3 mmol, 2.00 eq) with stirring at 25 ℃. The mixture was reacted at 80 ℃ for 16 hours with stirring under nitrogen protection, and the course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the temperature was reduced to 25 ℃, insoluble materials were removed by filtration, the filter cake was washed with acetonitrile (10 ml x 3), and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 35-1 (colorless oil, 2.84 g, 64% yield). MS (ESI, m/z) 142.1[ m ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ5.96–5.82(m,1H),5.24–5.14(m,1H),5.14–5.06(m,1H),3.67–3.59(m,1H),3.46–3.36(m,2H),3.16–3.07(m,1H),2.98–2.88(m,1H),2.81(s,1H),2.69–2.59(m,1H),2.37–2.26(m,1H),1.97–1.83(m,1H),1.83–1.73(m,1H),1.77–1.66(m,2H)。

Step 2

At zero degrees centigrade, nitrogen protectionTo a solution of compound 16-3a (250 mg, 0.387 mmol, 1.0 eq) and compound 35-1 (57.5 mg, 0.387 mmol, 1 eq) in tetrahydrofuran (3 ml) was added dropwise a solution of sodium tert-butoxide in tetrahydrofuran (2 mol/l, 194 ml, 0.387 mmol, 1 eq) with stirring. The resulting mixture was stirred at zero degrees centigrade for an additional 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, 10 ml of water is added into the reaction liquid for quenching reaction, then the mixed liquid is extracted by ethyl acetate (10 ml x 3), organic phases are combined, the organic phases are dried by anhydrous sodium sulfate, drying agents are removed by filtration, and the filtrate is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 5% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and give compound 35-2 (yellow oil, 175 mg, 59% yield). MS (ESI, m/z) 718.2/720.2[ 2 ], [ M + H ]] ⁺ 。

Step 3

Compound 35-2 (90 mg, 0.12 mmol, 1.0 eq), 1, 3-dimethylbarbituric acid (29 mg, 0.178 mmol, 1.5 eq), tetrakis (triphenylphosphine) palladium (7.24 mg, 0.006 mmol, 0.05 eq) and dichloromethane (2 ml) were added sequentially to a 50 ml three-necked flask under nitrogen protection at 25 ℃. The resulting mixture was reacted at 25 ℃ for 5 hours, the course of the reaction being monitored by chromatography on liquid and thin-layer chromatography. After the reaction is finished, the excessive solvent is removed by decompression and concentration to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 10% → 50% acetonitrile/water mobile phase (0.1% formic acid) over 20 minutes, detector UV254 nm to give compound 35-3 (orange solid, 74 mg, 85% yield). MS (ESI, m/z) 678.3/680.3[ M + H ] ] ⁺ 。

Step 4

To a solution of compound 35-3 (74.0 mg, 0.10 mmol, 1.00 eq), acetic acid (13.1 mg, 0.21 mmol, 2.00 eq) and ethyl glyoxylate (16.7 mg, 0.15 mmol, 1.50 eq) in methanol (4.0 ml) was added sodium cyanoborohydride (10.3 mg, 0.15 mmol, 1.50 eq) with stirring at 0 ℃. The mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was concentrated under reduced pressure to give a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 5% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 35-4 (white solid, 40 mg, 51% yield). MS (ESI, m/z) 764.3/766.3[ M ] +H] ⁺ 。

Step 5

To a mixture solution of compound 35-4 (40 mg, 0.031 mmol, 1.00 eq) in tetrahydrofuran/methanol/water (3/1, 1 ml) was added lithium hydroxide dihydrate (2.77 mg, 0.062 mmol, 2.00 eq) with stirring at 25 ℃. The mixture was stirred at 25 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was concentrated under reduced pressure to give a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 10% → 95% acetonitrile/water mobile phase (0.1% trifluoroacetic acid) over 20 minutes; a detector: UV254/220 nm; compound 35-5 was obtained (white solid, 15 mg, 61% yield). MS (ESI, m/z) 736.3/738.3[ 2 ] M + H ] ⁺ 。

Step 6

To compound 35-5 (14 mg, 0.015 mmol, 1.00 eq) was added N, N with stirring at 25 degrees celsiusTo a solution of dimethylformamide (1.00 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (9.13 mg, 0.022 mmol, 1.50 eq). The mixture was stirred at 25 ℃ for 20 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group was added to the reaction mixture]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (8.27 mg, 0.018 mmol, 1.20 equivalents) and N, N-diisopropylethylamine (8.28 mg, 0.060 mmol, 4.00 equivalents). The mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was purified by preparative high performance liquid chromatography: column XBridge Prep C18 OBD Column,19x150 mm, 5 micron; mobile phase A: water (10 mmol/l ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was carried out with 54% → 80% B phase in 8 minutes, detector UV 225/278 nm. Compound 35-6 was obtained (white solid, 14 mg, 76% yield). MS (ESI, m/z) 1148.6/1150.6[ 2 ] M + H ] ⁺ 。

Step 7

To a solution of compound 35-6 (14 mg, 0.012 mmol, 1.00 eq) in methanol (1 ml) was added dropwise a solution of hydrochloric acid in 1, 4-dioxane (4 mol/l, 1 ml) with stirring at 25 ℃. The reaction solution was reacted at 25 ℃ for 2 hours, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. The crude product obtained is purified by reverse phase chromatography (C18 column) eluting within 10 minutes with a mobile phase of 10% → 60% acetonitrile/water (0.1% hydrochloric acid); a detector: UV 254/220 nm; compound 35 (yellow solid, 9 mg, 69% yield) was obtained. MS (ESI, m/z): 1004.6/1006.1[ m ] +H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ9.04–8.98(m,1H),8.78–8.71(m,1H),8.03–7.96(m,1H),7.81(d,J＝8.5Hz,1H),7.48–7.34(m,5H),7.31(d,J＝2.4Hz,1H),7.26–7.14(m,2H),7.09–7.04(m,1H),4.79–4.62(m,2H),4.61–4.47(m,3H),4.45–4.28(m,4H),4.27–4.11(m,4H),4.09–3.98(m,1H),3.96–3.85(m,2H),3.79–3.64(m,2H),3.30–3.17(m,1H),2.43(s,3H),2.32–2.19(m,1H),2.10–1.82(m,9H),1.30–1.13(m,2H),0.98–0.72(m,10H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-121.85。

Example 36

((3S or 3R,7aS or 7 aR) -7a- (((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) hexahydro-1H-pyrrolin-3-yl) methyl (2- (3- ((S) -1- ((2S, 4R) -4-hydroxy-2- ((4- (4-methylthiazol-5-yl) benzyl) carbamoyl) pyrrolidin-1-yl) -3, 3-dimethyl-1-oxobutan-2-yl) amino) -3-oxopropoxy) ethyl) (methyl) carbamate diformate 36

Step 1

To a 500 ml round bottom flask were added N-methyl-2-hydroxyethylamine (10.0 g, 126.48 mmol, 1.0 eq), dichloromethane (60.0 ml), triethylamine (55.52 ml, 379.44 mmol, 3.0 eq) and di-tert-butyl dicarbonate (30.51 g, 132.80 mmol, 1.05 eq) sequentially with stirring at 25 ℃. The mixture was stirred at 25 ℃ for 3 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 36-1 (yellow oil, 17.0 g, 73% yield). MS (ESI, m/z) 176.1[ m + H ] ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ3.77–3.73(m,2H),3.42–3.38(m,2H),2.93(s,3H),1.47(s,9H)。

Step 2

To a solution of compound 36-1 (17.0 g, 92.2 mmol, 1.0 eq), 4-dimethylaminopyridine (1.2 g, 9.2 mmol, 0.1 eq) and triethylamine (19.6 g, 184.3 mmol, 2.0 eq) in dichloromethane (200.0 ml) was added p-toluenesulfonyl chloride (20.3 g, 101.4 mmol, 1.1 eq) at 0 ℃ with stirring under nitrogen. The resulting mixture was stirred at 25 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 60% methyl tert-butyl ether/petroleum ether mobile phase gradient and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 36-2 (yellow solid, 9.9 g, 26% yield). MS (ESI, m/z) 330.0[ m ] +H] ⁺ 。

Step 3

To a solution of 3-buten-1-ol (1.8 g, 23.8 mmol, 1.1 eq) in N-N dimethylformamide (10.0 ml) was added sodium hydride (60%) (1.1 g, 28.56 mmol, 1.2 eq) at 0 ℃ with stirring under nitrogen protection. The resulting mixture was stirred at 0 ℃ for 30 minutes under nitrogen protection. Then 36-2 (8.9 g, 21.6 mmol, 1.0 equiv) was added to the reaction. The mixture was allowed to react at 25 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, 300 ml of water was added to quench the reaction, and the aqueous layer was extracted with ethyl acetate (300 ml. Times.3). Combining the organic phases, drying the organic phases with anhydrous sodium sulfate, and filtering to remove the drying agent; and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 40% methyl t-butyl ether/petroleum ether mobile phase gradient, the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure, Compound 36-3 (colorless oil, 570 mg, 10.9% yield) was obtained. MS (ESI, m/z) 230.1[ m ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ5.86–5.76(m,1H),5.13–5.01(m,2H),3.55–3.47(m,4H),3.39–3.36(m,2H),2.91(s,3H),2.36–2.29(m,2H),1.46(s,9H)。

Step 4

To a solution of compound 36-3 (500.0 mg, 2.1 mmol, 1.0 eq) in methanol (3.0 ml) was added dropwise a solution of 1, 4-dioxane (4 mol/l, 3.0 ml) of hydrochloric acid with stirring at 0 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, excess solvent was removed by concentration under reduced pressure to give compound 36-4 (yellow liquid, 330.0 mg, 96% yield). MS (ESI, m/z) 130.2[ m ] +H] ⁺ 。

Step 5

Thionyl chloride (27.48 g, 228.67 mmol, 3.0 equiv.) is added dropwise to a solution of N-benzyloxycarbonyl-L-proline (20 g, 76.22 mmol, 1.0 equiv.) in methanol (200.0 ml) at 0 degrees celsius with stirring under nitrogen protection. The mixture was stirred at 0 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the filtrate is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 30% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 36-5 (colorless oil, 20 g, 94% yield). MS (ESI, m/z) 264.3[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.40–7.28(m,5H),5.22–5.00(m,2H),4.41–4.33(m,1H),3.80–3.41(m,5H),2.31–2.13(m,1H),2.09–1.73(m,3H)。

Step 6

To a solution of compound 36-5 (20 g, 72.17 mmol, 1.0 eq) in tetrahydrofuran (200.0 ml) was added slowly, under stirring at-78 ℃ under nitrogen protection, in succession lithium bistrimethylsilylamide (1 mol/l in tetrahydrofuran, 144 ml, 144 mmol, 2.0 eq) and 4-bromo-1-butene (12.3 g, 80.604 mmol, 1.2 eq). The mixture was stirred at 25 ℃ for 16 h and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was quenched by adding water (300 ml), followed by extraction with ethyl acetate (300 ml × 3), the organic phases were combined, washed with saturated brine (80 ml × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 30% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 36-6 (colorless oil, 17 g, yield 69%). MS (ESI, m/z) 318.1[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.42–7.29(m,5H),5.90–5.67(m,1H),5.21–4.90(m,4H),3.88–3.67(m,3H),3.57–3.44(m,2H),2.53–2.19(m,1H),2.18–1.79(m,7H)。

Step 7

To a solution of compound 36-6 (17 g, 50.884 mmol, 1.0 eq) in dichloromethane (200.0 ml) was slowly added m-chloroperoxybenzoic acid (23.12 g, 127.21 mmol, 2.5 eq) with stirring at 25 degrees celsius under nitrogen. The mixture was stirred at 25 ℃ for 5 hours and the progress of the reaction was monitored by thin layer chromatography (ethyl acetate/petroleum ether =1/4, r) _f = 0.2). After completion of the reaction, the reaction was quenched by adding saturated aqueous sodium thiosulfate (300.0 ml), followed by extraction with dichloromethane (300 ml. Times.3), and the reaction mixture was combinedAnd drying the organic phase with anhydrous sodium sulfate, filtering, and concentrating the filtrate to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 50% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 36-7 (tan oil, 12 g, 66% yield). MS (ESI, m/z) 334.1[ 2 ] M + H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.40–7.31(m,5H),5.20–5.06(m,2H),3.86–3.71(m,3H),3.55–3.44(m,2H),2.98–2.68(m,2H),2.56–2.23(m,2H),2.19–1.81(m,5H),1.72–1.35(m,2H)。

Step 8

To a solution of compound 36-7 (12 g, 34.195 mmol, 1.0 eq) in methanol (120.0 ml) was added palladium on carbon (10% palladium content, 1.2 g) with stirring at 25 ℃ under nitrogen. The nitrogen gas was replaced with hydrogen gas (1.5 atm) by a replacement gas operation. The mixture was stirred for 5 hours at 25 ℃ under hydrogen atmosphere and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, insoluble matter was removed by filtration, the cake was washed with methanol (100 ml), and the filtrate was concentrated under reduced pressure to obtain a crude product of compound 36-8. (colorless oil, 7 g, 97% yield). MS (ESI, m/z) 200.1[ 2 ] M + H] ⁺ 。

Step 9

To a solution of compound 36-8 (7 g, 33.375 mmol, 1.0 eq) and imidazole (9.09 g, 126.845 mmol, 3.8 eq) in N, N-dimethylformamide (70.0 ml) was slowly added tert-butyldiphenylchlorosilane (19.31 g, 66.75 mmol, 2.0 eq) with stirring at 0 degrees celsius under nitrogen protection. The mixture was stirred at 25 ℃ for 16 h and the reaction was carried out by thin layer chromatography (ethyl acetate/petroleum ether =1/3, r) _f =0.5,0.2). After the reaction is completed, the reaction is carried out in the reverse directionThe reaction was quenched by the addition of water (500 ml) and extracted with ethyl acetate (500 ml x 3), the organic phases were combined, washed with saturated brine (80 ml x 3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 40% ethyl acetate/petroleum ether mobile phase gradient and the fractions obtained were rotary evaporated under reduced pressure to remove the solvent and give the two compounds respectively. The relatively less polar compound was 36-8a (colorless oil, 6 g, 38% yield) and the relatively more polar compound was 36-8b (colorless oil, 5.7 g, 37% yield).

Compound 36-8b] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.74–7.63(m,4H),7.51–7.35(m,6H),4.03–3.96(m,1H),3.88–3.81(m,1H),3.74(s,3H),3.46–3.35(m,1H),3.01–2.79(m,2H),2.55–2.45(m,1H),2.30–2.15(m,1H),2.04–1.75(m,5H),1.70–1.53(m,1H),1.07(s,9H)。

Step 10

Compound 36-8b (5.6 g, 12.156 mmol, 1.0 eq) was added slowly to a solution of lithium aluminum hydride (1.46 g, 36.468 mmol, 3.0 eq) in tetrahydrofuran (56 ml) with stirring at-20 degrees celsius under nitrogen. The mixture was stirred at-20 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, water (1.46 ml), a 15% aqueous solution of sodium hydroxide (1.46 ml) and water (5.38 ml) were added to the reaction mixture in this order under stirring at zero degrees centigrade. The insoluble material was removed by filtration, the filter cake was washed with tetrahydrofuran (20 ml x 3), and the filtrate was concentrated under reduced pressure to give compound 36-9b (off-white solid, 5.0 g, 95% yield). MS (ESI, m/z) 410.3[ m + H ] ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.74–7.64(m,4H),7.49–7.33(m,6H),3.97–3.89(m,1H),3.79–3.72(m,1H),3.32–3.22(m,2H),3.22–3.13(m,1H),2.86–2.79(m,1H),2.74–2.65(m,1H),1.99–1.91(m,1H),1.82–1.57(m,6H),1.56–1.47(m,1H),1.06(s,9H)。

Step 11

Chiral resolution of compound 36-9b (5 g) from step 10 by supercritical liquid chromatography: chiral column (R, R) -WHELK-O1-Kromasil,3 × 25 cm, 5 μm; a mobile phase A: supercritical carbon dioxide, mobile phase B: isopropanol/dichloromethane =2/1 (2 moles/liter ammonia methanol); flow rate: 70 ml/min; elution was performed with 55% phase B in 6.5 min, detector UV 204 nm. Two products were obtained. The product of shorter residence time (3.27 min) was compound 36-9ba, ((3R or 3S,7aR or 7 aS) -3- ((tert-butyldiphenylsiloxy) methyl) tetrahydro-1H-pyrrolin-7 a (5H) -yl) methanol (colorless oil, 2.2 g, recovery 44%), MS (ESI, m/z): 410.3 2 [ +H ] +] ⁺ (ii) a The product with longer retention time (5.87 min) was compound 36-9bb, ((3S or 3R,7aS or 7 aR) -3- ((tert-butyldiphenylsiloxy) methyl) tetrahydro-1H-pyrrolin-7 a (5H) -yl) methanol (colorless oil, 1.9 g, recovery 38%), MS (ESI, m/z): 410.3[ M ] +H ])] ⁺ 。

Step 12

To a solution of compound 16-3a (1.36 g, 2.106 mmol, 1.00 equiv) in anhydrous tetrahydrofuran (10 ml) was added dropwise, under stirring at 0 ℃ under nitrogen protection, compound 36-9ba (1.00 g, 2.317 mmol, 1.10 equiv) and potassium tert-butoxide in tetrahydrofuran (1.0 mol/l, 2.5 ml, 2.527 mmol, 1.20 equiv) in that order. The reaction solution reacts for 1 hour under the protection of nitrogen at 0 ℃, and the reaction process is monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. The crude product was purified by silica gel chromatography eluting with 0% → 10% methanol/dichloromethane mobile phase and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford 36-10ba (yellow solid, 1.09 g, 50% yield) 。MS(ESI,m/z):986.3/988.3[M+H] ⁺ 。

Step 13

Tetrabutylammonium fluoride (1.59 g, 5.778 mmol, 6.00 equiv.) is added dropwise to a solution of compound 36-10ba (1.00 g, 0.963 mmol, 1.00 equiv.) in tetrahydrofuran (8 ml) with stirring at 25 ℃. The reaction solution was reacted at 25 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. The crude product was purified by silica gel chromatography eluting with 0% → 10% methanol/dichloromethane mobile phase and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 36-11ba (white solid, 750 mg, 98% yield). MS (ESI, m/z) 748.2/750.2[ m ] +H] ⁺ 。

Step 14

To a schlenk tube were added sequentially compound 36-11ba (200.0 mg, 0.25 mmol, 1.0 eq), tetrahydrofuran (3.0 ml), p-nitrophenyl chloroformate (107.7 g, 0.5 mmol, 2.0 eq), compound 36-4 (103.6 mg, 0.7 mg, 3.0 eq) and N-diisopropylethylamine (138.1 mg, 1.0 mmol, 4.0 eq) at 25 degrees celsius with stirring under nitrogen protection. The resulting mixture was stirred at 0 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating under reduced pressure to remove redundant reagents to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fractions were freed of solvent by rotary evaporation under reduced pressure to give compounds 36-12ba (white solid, 120.0 mg, 49% yield). MS (ESI, m/z) 903.1/905.1[ m + H ] ] ⁺ 。

Step 15

To a mixed solution of compounds 36-12ba (110.0 mg, 0.1 mmol, 1.0 eq) and sodium periodate (100.1 mg, 0.5 mmol, 4.0 eq) in carbon tetrachloride/acetonitrile/water (2/2/3, 4.5 ml) was added ruthenium trichloride (1.3 mg, 0.005 mmol, 0.05 eq) at 0 ℃ with stirring under nitrogen protection. The mixture was stirred at 0 ℃ for 3 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The resulting mixture was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 90% acetonitrile/water mobile phase (0.1% ammonium bicarbonate) over 25 minutes; detector, UV254 nm; compound 36-13ba (white solid, 55 mg, 49% yield) MS (ESI, m/z): 921.5/923.5[ 2 ] M + H] ⁺ 。

Step 16

To a solution of compound 36-13ba (50 mg, 0.05 mmol, 1.0 eq) in N, N-dimethylformamide (2.00 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethylurea hexafluorophosphate (23.7 mg, 0.06 mmol, 1.2 eq) with stirring at 25 ℃. The mixture was stirred at 25 ℃ for 20 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group was added to the reaction mixture ]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] carbonyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (26.9 mg, 0.06 mmol, 1.2 eq) and N, N-diisopropylethylamine (26.9 mg, 0.2 mmol, 4.0 eq). The mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the mixture was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 90% acetonitrile/water mobile phase (0.1% trifluoroacetic acid) over 25 minutes; detector UV254 nm; to give compound 36-14ba (white solid)Body, 46 mg, 63% yield). MS (ESI, m/z) 1333.0/1335.0[ m + H ]] ⁺ 。

Step 17

To a solution of compound 36-14ba (45.0 mg, 0.03 mmol, 1.0 eq) in methanol (1 ml) was added dropwise a solution of hydrochloric acid in 1, 4-dioxane (4 mol/l, 1 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Purifying the obtained crude product by high performance liquid chromatography with a chromatographic column Sunfire prep C18 column,30x150 mm, 5 micron; mobile phase A: water (0.05% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution with 10% → 35% B phase over 7 minutes, detector UV 220/235 nm gave compound 36 (white solid, 35 mg, 79% yield). MS (ESI, m/z) 1189.1/1191.1[ 2 ] M + H ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.98–8.91(m,1H),8.48–8.34(m,2H),8.03–7.97(m,1H),7.95–7.81(m,2H),7.51–7.34(m,6H),7.32–7.18(m,2H),7.09(d,J＝2.4Hz,1H),4.56–4.40(m,7H),4.33–4.21(m,3H),4.02–3.90(m,2H),3.86–3.58(m,7H),3.55–3.46(m,2H),3.43–3.29(m,2H),3.28–3.18(m,1H),3.15–3.05(m,2H),2.91–2.81(m,3H),2.48–2.35(m,5H),2.24–1.78(m,14H),1.26–1.16(m,2H),1.00–0.83(m,9H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-122.36。

Example 37

(2S, 4R) -1- ((S) -2- (3- (3- ((3R, 5S) -5- (((S or R) -4- ((1R, 5S) -3, 8-diazabicyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) -1-methylpyrrolidin-3-yl) oxy) propionamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide dihydrochloride 37

Step 1

To a solution of (2s, 4r) -1- (tert-butoxycarbonyl) -4-hydroxy-2- (hydroxymethyl) pyrrolidine (10.0 g, 43.7 mmol, 1.0 eq) and imidazole (3.3 g, 45.9 mmol, 1.1 eq) in N, N-dimethylformamide (100 ml) was slowly added tert-butyldiphenylchlorosilane (13.3 g, 45.9 mmol, 1.1 eq) with stirring at 0 degrees celsius under nitrogen protection. The mixture was allowed to react for 16 hours at 25 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction was diluted with 600 ml of water at 0 ℃ and extracted with ethyl acetate (600 ml × 3), and the organic phases were combined, washed with saturated brine (200 ml × 3), and dried over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 40% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 37-1 (white solid, 10.5 g, 52% yield). MS (ESI, m/z) 456.2[ 2 ], [ M + H ] ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.69–7.62(m,4H),7.48–7.34(m,6H),4.59–4.51(m,1H),4.12–4.01(m,1H),4.01–3.77(m,1H),3.73–3.64(m,1H),3.63–3.47(m,2H),2.40–2.29(m,1H),2.13–2.00(m,1H),1.87–1.69(m,1H),1.41(s,9H),1.06(s,9H)。

Step 2

To a solution of compound 37-1 (5.65 g, 11.8 mmol, 1.5 eq) in N, N-dimethylformamide (30 ml) was added sodium hydride (60%, 0.5 g, 11.8 mmol, 1.5 eq) with stirring at 0 degrees celsius under nitrogen. After the mixture was stirred at 0 ℃ for 0.5 hour, it was addedCompound 25-3 (2.4 g, 7.9 mmol, 1.0 eq). The reaction solution was reacted at 25 ℃ for 16 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, 200 ml of water was added to the reaction solution at 0 ℃ to quench the reaction. The aqueous phase was extracted with ethyl acetate (200 ml x 3), the combined organic phases were washed with saturated brine (80 ml x 3), dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% methanol/water mobile phase (0.1% trifluoroacetic acid) over 25 minutes; detector UV254 nm; compound 37-2 was obtained (colorless oil, 1.7 g, 36% yield). MS (ESI, m/z) 568.3[ m + H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.67–7.60(m,4H),7.45–7.31(m,6H),5.92–5.76(m,1H),5.15–4.92(m,2H),4.15–3.90(m,2H),3.73–3.34(m,10H),2.39–2.20(m,3H),2.15–1.95(m,1H),1.88–1.76(m,2H),1.48–1.30(m,9H),1.05(s,9H)。

Step 3

Compound 37-2 (1.7 g, 2.8 mmol, 1.0 eq) was dissolved in tetrahydrofuran (17 ml) with stirring at 25 ℃, followed by the addition of tetrabutylammonium fluoride (1.5 g, 5.5 mmol, 2.0 eq). The reaction solution was reacted for 1.5 hours at 25 ℃ with stirring, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the reaction was diluted with 50 ml of water, the aqueous phase was extracted with ethyl acetate (50 ml x 3), and the organic phases were combined and dried over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 37-3 (colorless oil, 900 mg, 94% yield). MS (ESI, m/z) 330.1[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ5.88–5.76(m,1H),5.13–4.97(m,2H),4.10–4.01(m,1H),3.96–3.88(m,1H),3.73–3.67(m,1H),3.61–3.54(m,2H),3.53–3.44(m,6H),3.41–3.35(m,1H),2.37–2.28(m,2H),2.14–2.06(m,1H),1.85–1.77(m,2H),1.74–1.60(m,1H),1.47(s,9H)。

Step 4

To a solution of compound 37-3 (850 mg, 2.5 mmol, 1.0 eq) in anhydrous tetrahydrofuran (9 ml) was slowly added lithium aluminum hydride (195.9 mg, 4.9 mmol, 2.0 eq) with stirring at 0 degrees celsius under nitrogen blanket. The reaction solution was reacted at 55 ℃ for 2 hours, and the reaction process was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, water (195.9 mg), a 15% aqueous solution of sodium hydroxide (195.9 mg), and water (588 mg) were added to the reaction solution at 0 ℃. The insoluble material was removed by filtration, the filter cake was washed with tetrahydrofuran (15 ml x 3), and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and give compound 37-4 (colorless oil, 430 mg, yield 72%). MS (ESI, m/z) 244.2[ 2 ], [ M ] +H] ⁺ 。

Step 5

To a solution of compound 16-3a (250 mg, 0.39 mmol, 1.0 equiv.) and compound 37-4 (119.0 mg, 0.46 mmol, 1.2 equiv.) in anhydrous tetrahydrofuran (3 ml) was added a solution of potassium tert-butoxide in tetrahydrofuran (1 mol/l, 0.46 ml, 0.46 mmol, 1.2 equiv) under-20 ℃ nitrogen protection. The mixture was reacted at-20 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, 10 ml of water was added to the reaction solution to quench the reaction, followed by extraction with ethyl acetate (15 ml. Times.3), organic phases were combined and dried over anhydrous sodium sulfate, and filtered to remove impurities Dissolving, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 37-5 (white solid, 150 mg, 45% yield). MS (ESI, m/z): 820.3/822.3[ m ] +H] ⁺ 。

Step 6

Compound 37-5 (160 mg, 0.19 mmol, 1.0 eq) was dissolved in 1.7 ml of carbon tetrachloride and 1.7 ml of acetonitrile with stirring at 25 ℃, followed by the addition of sodium periodate (250.3 mg, 1.1 mmol, 6.0 eq). The system was cooled to 0 ℃ and then an aqueous solution (2.5 ml) of ruthenium trichloride (2.2 mg, 0.01 mmol, 0.05 eq) was slowly added dropwise. The reaction solution was reacted at 0 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was poured into 15 ml of water, the aqueous phase was extracted with a mixed solution of chloroform/isopropanol (3, 1, 20 ml × 3), and the organic phases were combined and dried over anhydrous sodium sulfate. Insoluble substances are removed by filtration, and the organic phase is concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% methanol/water mobile phase (0.1% ammonia) over 25 minutes; detector UV254 nm; compound 37-6 was obtained (white solid, 60 mg, 38% yield). MS (ESI, m/z) 838.4/840.4[ m ] +H ] ⁺ 。

Step 7

To a solution of compounds 37-6 (60 mg, 0.07 mmol, 1.0 eq) in N, N-dimethylformamide (3 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (32.7 mg, 0.08 mmol, 1.2 eq) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 15 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) was added to the reaction mixture]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (44.4 mg, 0.08 mmol, 1.2 eq) and N, N-diisopropylethylamine (46.3 mg, 0.34 mmol, 5.0 eq). The reaction solution was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was purified by high performance liquid chromatography, column xsselect CSH Prep C18 OBD,19 × 150 mm, 5 μm; mobile phase A: water (10 mmol/l ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was performed with 56% → 80% B phase over 8 minutes, detector UV 220/235 nm. Compound 37-7 was obtained (colorless oil, 53 mg, 59% yield). MS (ESI, m/z) 1264.5/1266.5[ m + H ] ] ⁺ 。

Step 8

To a solution of compound 37-7 (53 mg, 0.04 mmol, 1.0 eq) in methanol (2 ml) was added dropwise a solution of hydrochloric acid in 1, 4-dioxane (4 mol/l, 2 ml) with stirring at 0 ℃. The mixture was allowed to react for 2 hours at 25 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is decompressed and concentrated to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% methanol/water mobile phase (0.1% hydrochloric acid) over 25 minutes; detector, UV220 nm; compound 37 was obtained (yellow solid, 32.3 mg, yield 64%). MS (ESI, m/z) 1120.3/1122.3[ m ] +H] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ11.36–11.14(m,1H),10.17–9.97(m,1H),9.92–9.73(m,1H),9.14–9.06(m,1H),8.45(d,J＝7.8Hz,1H),8.04–7.98(m,1H),7.91–7.78(m,2H),7.48–7.41(m,3H),7.41–7.35(m,2H),7.33–7.28(m,1H),7.26–7.16(m,2H),7.14–7.09(m,1H),4.97–4.84(m,1H),4.84–4.74(m,2H),4.62–4.39(m,4H),4.30–4.25(m,1H),4.24–4.11(m,3H),4.04–3.91(m,3H),3.87–3.77(m,1H),3.66–3.51(m,4H),3.49–3.34(m,5H),3.22–3.13(m,1H),3.03–2.92(m,3H),2.46(s,3H),2.41–2.28(m,2H),2.09–1.85(m,6H),1.82–1.63(m,3H),1.51–1.31(m,3H),1.01–0.80(m,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-121.95。

Example 38

(2S, 4R) -1- ((S) -2- (3- (3- ((S) -2- ((4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -8-fluoro-7- (3-hydroxynaphthalen-1-yl) pyridine [4,3-d ] pyrimidin-2-yl) oxy) methyl) pyrrolidin-1-yl) propoxy) propionamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4-methylthiazole-5-benzyl) pyrrolidine-2-carboxamide diformate 38

Step 1:

to a solution of 2-chloro-3-fluoro-4-aminopyridine (1 g, 6.82 mmol, 1.0 eq) in acetonitrile (10 ml) was added N-iodosuccinimide (1.84 g, 7.77 mmol, 1.2 eq) and p-toluenesulfonic acid (130 mg, 0.65 mmol, 0.1 eq) with stirring at 25 ℃. The mixture was reacted at 70 ℃ for 16 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the mixture was cooled to 25 ℃, diluted with 30 ml of water, extracted with ethyl acetate (50 ml x 3) and the organic phases combined. The organic phase is washed successively with 50 ml of saturated sodium carbonate solution, 50 ml of saturated sodium sulfite solution and 50 ml of saturated brine, the organic phase is dried over anhydrous sodium sulfate, filtered and the filtrate is concentrated to give a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 45% ethyl acetate/petroleum ether and the fractions were freed of solvent by rotary evaporation under reduced pressure to give compound 38-1 (yellow solid, 1.72 g, 95% yield). MS (ESI, m/z) 272.9/274.9[ m + H ] ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.17(s,1H),4.84(s,2H)。

Step 2:

to a solution of compound 38-1 (1 g, 3.67 mmol, 1.0 eq) in ethanol (10 ml) was added triethylamine (1.38 g, 12.96 mmol, 3.6 eq) and bis (triphenylphosphine) palladium dichloride (266 mg, 0.36 mmol, 0.1 eq) with stirring at 25 degrees celsius under nitrogen protection. The mixture was reacted for 15 hours at 80 ℃ under an atmosphere of carbon monoxide, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, cooling the reaction liquid to 25 ℃, filtering the reaction liquid by using kieselguhr, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 31% ethyl acetate/petroleum ether and the fractions were freed of solvent by rotary evaporation under reduced pressure to give compound 38-2 (yellow solid, 660 mg, 74% yield). MS (ESI, m/z) 219.3/221.3[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ8.56(s,1H),4.41(q,J＝7.1Hz,2H),1.43(t,J＝7.1,3H)。

And step 3:

trichloroacetyl isocyanate (853 mg, 4.30 mmol, 1.5 equivalents) was added dropwise to a solution of compound 38-2 (660 mg, 2.87 mmol, 1.0 equivalent) in anhydrous tetrahydrofuran (6 ml) at 25 deg.c with stirring under nitrogen. The mixture was reacted at 25 ℃ for 20 minutes, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction solution to obtain a crude product. The crude product was slurried with methyl tert-butyl ether (10 ml), filtered, the filter cake washed with methyl tert-butyl ether (2 ml x 3), and the filter cake dried to give compound 38-3 (white solid, 1.0 g, 77% yield). MS (ESI, m/z): 406.0/408.0/410.0M + H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ11.19(s,1H),8.91(s,1H),8.78(d,J＝0.8Hz,1H),4.48(q,J＝7.1Hz,2H),1.43(t,J＝7.1,3H)。

And 4, step 4:

to a solution of compound 38-3 (1 g, 2.334 mmol, 1.00 eq) in methanol (10 ml) was added dropwise a solution of ammonia in methanol (7 mol per liter, 1 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction solution to obtain a crude product. The crude product was slurried with methyl tert-butyl ether (10 ml), filtered, the filter cake washed with methyl tert-butyl ether (2 ml x 3), and the filter cake dried to give compound 38-4 (white solid, 594 mg, 94% yield). MS (ESI, m/z) 216.1/218.1[ m ] +H] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ8.34(d,J＝1.2Hz,1H)。

And 5:

to a dry 100 ml single neck flask were added sequentially compound 38-4 (500 mg, 2.20 mmol, 1.0 eq), phosphorus oxychloride (9 ml) and N, N-diisopropylethylamine (0.9 ml) at 0 degrees celsius under nitrogen protection. The mixture was stirred at 0 ℃ for 10 minutes and then reacted at 90 ℃ for 12 hours, the course of the reaction being monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is decompressed and concentrated to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a gradient of 0% → 30% ethyl acetate/petroleum ether and the fractions were freed from the solvent by rotary evaporation under reduced pressure to give compound 38-5 as a yellow solid (425 mg, 72% yield).

Step 6:

compound 38-5 (425 mg, 1.68 mmol, 1.0 eq) was dissolved in 5 ml of dichloromethane with stirring at 25 degrees c under nitrogen. To this solution were added N, N-diisopropylethylamine (652 mg, 4.80 mmol, 3.0 eq.) and (1R, 5S) -3, 8-diazabicyclo [3.2.1]Octane-8-carboxylic acid tert-butyl ester (357 mg, 1.60 mmol, 1.0 eq) was then reacted at 25 ℃ for 1 hour, the course of which was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a gradient of 0% → 30% ethyl acetate/dichloromethane and the fractions were stripped of solvent by rotary evaporation under reduced pressure to give compound 38-6 (yellow solid, 700 mg, 97% yield). MS (ESI, m/z) 428.2/430.2[ 2 ], [ M + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ8.86(d,J＝0.6Hz,1H),4.59–4.41(m,4H),3.78–3.71(m,2H),2.04–1.96(m,2H),1.75–1.65(m,2H),1.54(s,9H)。

Step 7

To a solution of compound 38-6 (700 mg, 1.64 mol, 1.0 eq) and compound 25-4 (419 mg, 1.96 mmol, 1.2 eq) in anhydrous tetrahydrofuran (10.0 ml) was added a solution of potassium tert-butoxide in tetrahydrofuran (1 mol/l, 1.96 ml, 1.96 mmol, 1.2 eq) with stirring at 0 ℃. The resulting mixture was stirred at 0 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, 50 ml of water is added to quench the reaction, the aqueous phase is extracted by ethyl acetate (50 ml x 3), the organic phases are combined, the organic phases are washed by 50 ml of saturated saline solution, the washed organic phases are dried by anhydrous sodium sulfate, the drying agent is removed by filtration, and the filtrate is concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and give compound 38-7 (colorless oil, 570) Mg, 57% yield). MS (ESI, m/z) 605.4/607.4[ 2 ] M + H] ⁺ 。

Step 8

To a reaction flask were added sequentially the compounds 38-7 (550 mg, 0.86 mmol, 1.0 eq), 4- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) naphthalen-2-ol (368.28 mg, 1.29 mmol, 1.5 eq), cesium carbonate (592.25 mg, 1.73 mmol, 2.0 eq), tetrakistriphenylphosphine palladium (105.02 mg, 0.086 mmol, 0.1 eq), 1, 4-dioxane (5.0 ml) and water (1.0 ml) at 25 ℃ with stirring under nitrogen. The mixture was stirred at 100 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, 50 ml of dilution was added to the reaction solution, and the aqueous phase was extracted with ethyl acetate (50 ml × 3). The organic phases are combined, washed with 50 ml of saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate is concentrated under reduced pressure to give a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 60% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 38-8 (white solid, 550 mg, 85% yield). MS (ESI, m/z) 713.1[ 2 ] M + H ] ⁺ 。

Step 9

To a solution of compound 38-8 (480 mg, 0.64 mmol, 1.0 eq) and imidazole (137.52 mg, 1.92 mmol, 3.0 eq) in dichloromethane (5 ml) was added tert-butyldimethylchlorosilane (152.23 mg, 0.96 mmol, 1.5 eq) at 0 ℃ with stirring under nitrogen. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, 50 ml of water was added to the reaction solution, and extracted with dichloromethane (50 ml × 3). After combination haveThe organic phase was washed with 50 ml of saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to give a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 80% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 38-9 (white solid, 420 mg, 75% yield). MS (ESI, m/z) 827.4[ m + H ]] ⁺ 。

Step 10

To a mixed solution of compounds 38-9 (180 mg, 0.21 mmol, 1.0 eq) and sodium periodate (279.28 mg, 1.24 mmol, 6.0 eq) in acetonitrile (1.0 ml), water (1.5 ml) and carbon tetrachloride (1.0 ml) was added ruthenium chloride monohydrate (2.45 mg, 0.01 mmol, 0.05 eq) with stirring at 0 degrees celsius. The resulting mixture was stirred at 0 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the excessive solvent is removed by decompression and concentration to obtain a crude product. The resulting mixture was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% ammonia, 25% to 28% ammonia in aqueous ammonia) over 25 minutes; detector, UV254 nm; compound 38-10 was obtained (yellow oil, 86 mg, 47% yield). MS (ESI, m/z) 845.6[ m + H ] ] ⁺ 。

Step 11

(2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) -amide was added to a solution of 38-10 (81 mg, 0.091 mmol, 1.0 eq) and 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (43.73 mg, 0.11 mmol, 1.2 eq) in N, N-dimethylformamide (1 ml) with stirring at 25 deg.C]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Methyl radical]Pyrrolidine-2-carboxylic acid estersAmine hydrochloride (53.72 mg, 0.11 mmol, 1.2 eq) and N, N-diisopropylethylamine (49.55 mg, 0.364 mmol, 4.0 eq). The resulting mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the resulting mixture was purified by reverse phase flash chromatography (C18 column) using 5% → 95% of [ acetonitrile/methanol (3/1) ] in 30 minutes]Elution is carried out on water mobile phase (0.1 percent of ammonia water, the ammonia content of the ammonia water is 25 to 28 percent); detector UV254 nm; compound 38-11 was obtained (yellow solid, 82 mg, 68% yield). MS (ESI, m/z) 1257.8[ 2 ], [ M ] +H] ⁺ 。

Step 12

Tetrabutylammonium fluoride (51.14 mg, 0.186 mmol, 3.0 equiv.) is added to a solution of compounds 38-11 (82 mg, 0.062 mmol, 1.0 equiv.) in tetrahydrofuran (3 ml) at 25 ℃ with stirring under nitrogen. The mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the crude product was obtained by concentration under reduced pressure. The crude product was purified by reverse phase flash chromatography (C18 column) over 30 minutes with 5% → 95% of [ acetonitrile/methanol (1/1) ] ]Eluting water mobile phase (0.1% ammonia water, ammonia content of 25% -28%) in ammonia water; detector UV254 nm; compound 38-12 was obtained (yellow solid, 42 mg, 59% yield). MS (ESI, m/z) 1143.7[ m + H ]] ⁺ 。

Step 13

To a solution of compound 38-12 (40 mg, 0.33 mmol, 1.0 eq) in dichloromethane (2 ml) was added trifluoroacetic acid (1 ml) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the crude product was obtained by concentration under reduced pressure. CoarseThe product was purified by reverse phase flash chromatography (C18 column) in 30 minutes with 5% → 95% of [ acetonitrile/methanol (1/1)]Elution with aqueous mobile phase (0.1% formic acid); detector UV254 nm; compound 38 was obtained (white solid, 25 mg, 63% yield). MS (ESI, m/z) 1043.5[ m + H ]] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ9.16(s,1H),8.97(s,1H),8.59–8.53(m,1H),8.20(s,2H),7.86(d,J＝9.4Hz,1H),7.79(d,J＝8.3Hz,1H),7.55(d,J＝8.4Hz,1H),7.46–7.34(m,5H),7.30–7.21(m,3H),4.55–4.32(m,7H),4.25–4.18(m,1H),4.15–4.09(m,1H),3.71–3.47(m,9H),3.42–3.34(m,2H),3.07–3.01(m,1H),2.96–2.78(m,2H),2.59–2.52(m,1H),2.43(s,3H),2.40–2.25(m,2H),2.22–2.14(m,1H),2.07–1.99(m,1H),1.93–1.85(m,2H),1.74–1.61(m,9H),0.89(s,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-139.66。

Example 39

(2S, 4R) -1- ((S) -2- (2- (4- (2- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-ylethyl) -1H-1,2, 3-triazol-1-yl) acetamido-3-dimethylbutyryl) -4-hydroxy-4-benzyl-4-thiazole-2-carboxamide-2-benzyl-carboxamide dihydrochloride 39

Step 1

To a mixed solution of ethyl 2-azidoacetate (5 g, 36.788 mmol, 1 eq) in water (25.0 ml)/tert-butanol (25.0 ml) was added sodium L-ascorbate (1.70 g, 8.093 mmol, 0.22 eq), copper sulfate (0.68 g, 4.047 mmol, 0.11 eq) and 3-butyn-1-ol (4.07 g, 55.182 mmol, 1.5 eq) at 25 ℃ with stirring. The mixture is stirred and reacted for 2 hours at the temperature of 25 ℃, and the reaction process is carried out by liquid chromatography and thin layer chromatographyTo monitor. After completion of the reaction, the reaction was quenched by addition of 100 ml of water at 25 ℃ and the aqueous phase was extracted with chloroform/isopropanol (3/1,100 ml. Times.3). The combined organic layers are washed by 400 ml of saturated sodium chloride solution, the organic phase after washing is dried by anhydrous sodium sulfate, the drying agent is removed by filtration, and the filtrate is decompressed and concentrated to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 7% methanol/dichloromethane and the resulting fraction was freed of solvent by rotary evaporation under reduced pressure to give compound 39-1 (white oil, 5.04 g, 65%). MS (ESI, m/z) 200.1[ 2 ] M + H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.56(s,1H),5.14(s,2H),4.26(q,J＝7.1Hz,2H),3.92(t,J＝6.0Hz,2H),3.07(s,1H),2.97(t,J＝6.0Hz,2H),1.30(t,J＝7.1Hz,3H)。

Step 2

To a solution of compound 39-1 (2 g, 9.538 mmol, 1.00 eq), triethylamine (3.05 g, 28.614 mmol, 3 eq) and 4-dimethylaminopyridine (0.12 g, 0.954 mmol, 0.1 eq) in dichloromethane (30.0 ml) was added p-toluenesulfonyl chloride (2.23 g, 11.446 mmol, 1.2 eq) at 0 ℃ with stirring under nitrogen. The reaction solution was stirred at 25 ℃ for 16 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction was quenched by the addition of 50 ml of water at 0 ℃, the aqueous phase was extracted with dichloromethane (50 ml x 3), and the combined organic phases were dried over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a 0% → 30% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 39-2 (yellow oil, 2.98 g, 83% yield). MS (ESI, m/z) 354.0[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.79–7.71(m,2H),7.57(s,1H),7.33(d,J＝8.1Hz,2H),5.13(s,2H),4.33–4.23(m,4H),3.13(t,J＝6.5Hz,2H),2.45(s,3H),1.31(t,J＝7.1Hz,3H)。

Step 3

To a solution of compound 39-2 (1 g, 2.68 mmol, 1.0 eq) and L-prolinol (314.85 mg, 2.957 mmol, 1.10 eq) in acetonitrile (5.0 ml) was added potassium carbonate (782.17 mg, 5.376 mmol, 2.0 eq) with stirring at 25 ℃. The mixture was stirred at 60 ℃ for 3 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, insoluble matter was removed by filtration, the cake was washed with acetonitrile (50 ml. Times.3), and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 7% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 39-3 (white solid, 320 mg, 40% yield). MS (ESI, m/z) 283.2[ m ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.51(s,1H),5.13(s,2H),4.31–4.21(m,2H),3.70–3.58(m,1H),3.46–3.36(m,1H),3.35–3.24(m,1H),3.23–3.12(m,1H),3.09–2.94(m,2H),2.80–2.67(m,2H),2.46–2.32(m,1H),1.97–1.85(m,1H),1.83–1.73(m,3H),1.30(t,J＝7.1Hz,3H)。

Step 4

To a solution of compound 16-3a (300 mg, 0.464 mmol, 1.0 eq) and compound 39-3 (151.9 mg, 0.511 mmol, 1.1 eq) in dichloromethane (5.0 ml) was added a solution of potassium tert-butoxide in tetrahydrofuran (1 mol per liter, 1.16 ml, 1.16 mmol, 2.5 eq) at-15 ℃ with stirring under nitrogen. The reaction solution was stirred at-15 ℃ for 4 hours and the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, 2 ml of water is added into the system for quenching reaction, the reaction is concentrated under reduced pressure to obtain a crude product, and the obtained crude product is directly purified by a reversed phase flash chromatography column (C18 column) and is used for 30% → 80% in 30 minutes (acetonitrile/methanol = 1/1)/aqueous mobile phase (0.1% ammonium bicarbonate) was eluted; detector UV220/254 nm; compound 39-4 was obtained (white solid, 112.0 mg, 27% yield). MS (ESI, m/z) 831.4/833.4[ 2 ] M + H] ⁺ 。

Step 5

To a solution of compound 39-4 (100.0 mg, 0.11 mmol, 1.0 eq) in N, N-dimethylformamide (1.5 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (54.88 mg, 0.137 mmol, 1.2 eq) with stirring at room temperature. The mixture was further stirred at 25 ℃ for 10 minutes, and (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group was added to the reaction mixture]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] carbonyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (61.80 mg, 0.125 mmol, 1.1 eq) and N, N-diisopropylethylamine (62.19 mg, 0.456 mmol, 4 eq). The mixture was stirred at 25 ℃ for a further 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin-layer chromatography. After the reaction was complete, the resulting mixture was purified by preparative high performance liquid chromatography on Column XBridge Prep C18 OBD Column 19x150 mm, 5 μm; mobile phase A: water (0.05% ammonium bicarbonate), mobile phase B: methanol; flow rate: 25 ml/min; elution was performed with 83% → 84% B phase in 6 minutes, detector UV220 nm. Compound 39-5 was obtained (white solid, 85 mg, 55% yield). MS (ESI, m/z) 1243.1/1245.1[ m ] +H ] ⁺ 。

Step 6

To a solution of compound 39-5 (75.0 mg, 0.06 mmol, 1 eq) in methanol (2.0 ml) was added a solution of 1, 4-dioxane (4 mol per liter, 2.0 ml) of hydrochloric acid with stirring at 0 ℃. The resulting mixture was at 25 deg.CThe reaction was stirred for 1.5 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove excess reagents, and the resulting mixture was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 95% acetonitrile/water mobile phase (0.1% hydrochloric acid) over 30 minutes; detector UV220/254 nm; compound 39 was obtained (yellow solid, 54.8 mg, 77% yield). MS (ESI, m/z): 1099.1/1101.1[ M ] +H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ11.20–10.98(m,1H),10.14–9.93(m,1H),9.86–9.66(m,1H),9.19–9.09(m,1H),8.72–8.61(m,1H),8.55(d,J＝9.1Hz,1H),8.04–7.97(m,1H),7.97–7.90(m,1H),7.81(d,J＝8.3Hz,1H),7.50–7.36(m,5H),7.34–7.28(m,1H),7.28–7.17(m,2H),7.12(d,J＝2.3Hz,1H),5.25–5.19(m,2H),4.79–4.70(m,2H),4.62–4.39(m,6H),4.37–4.30(m,1H),4.29–4.09(m,4H),4.07–3.91(m,3H),3.85–3.61(m,3H),3.60–3.53(m,1H),3.52–3.41(m,1H),3.31–3.10(m,3H),2.47(s,3H),2.33–2.20(m,1H),2.13–1.84(m,9H),0.96(s,9H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-121.84。

Example 40

(2S, 4R) -1- ((S) -2- (3- (3- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl) -6, 8-difluoroquinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) propoxy) propanamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide diformate 40

Step 1

Reference was made to the synthesis of compound 38-6 using 3-chloro-2, 4-difluoroaniline instead of 2-chloro-3-fluoro-4-aminopyridine to give compound 40-1 (a pale yellow solid). MS (ESI, m/z) 489.1/491.0[ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.44–7.36(m,1H),4.48–4.31(m,4H),3.75–3.54(m,2H),2.08–1.92(m,2H),1.79–1.70(m,2H),1.54(s,9H)。

Step 2

To a solution of compound 40-1 (2.5 g, 4.849 mmol, 1 eq), cesium carbonate (4.99 g, 14.547 mmol, 3 eq), and triethylenediamine (0.11 g, 0.970 mmol, 0.2 eq) in N, N-dimethylformamide (25 ml) at 25 degrees celsius under stirring under nitrogen, compound 25-4 (1.31 g, 5.819 mmol, 1.2 eq) was added. The mixture was stirred at 90 ℃ for 3 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, 200 ml of water was added to dilute the reaction solution, the aqueous layer was extracted with ethyl acetate (300 ml × 3), and after combining, the organic layer was washed with saturated brine (100 ml × 3) and dried over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 40-2 (yellow oil, 1.13 g, 29% yield). MS (ESI, m/z) 666.1/667.9[ M + H ]] ⁺ 。

Step 3

To a mixed solution of compound 40-2 (800 mg, 1.14 mmol, 1.0 equivalent), compound 40-3 (518.78 mg, 1.368 mmol, 1.2 equivalent, synthesized in reference to patent (WO 2021041671 (A1)), 2-dicyclohexylphosphine-2 ',4',6' -triisopropylbiphenyl (114.42 mg, 0.228 mmol, 0.2 equivalent), and tris (dibenzylideneacetone) dipalladium (109.90 mg, 0.114 mmol, 0.1 equivalent) in 1, 4-dioxane/water (20 ml/5 ml) was added cesium carbonate (782.04 mg, 2.280 mmol) in portions under stirring at 25 degrees celsius under nitrogen protection Moles, 2 equivalents). The mixture was reacted at 80 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature, and the mixture was concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography, eluting with a methanol/dichloromethane mobile phase gradient of 0% → 6%, removing the solvent from the resulting fraction by rotary evaporation under reduced pressure, and purifying the crude product by high performance liquid chromatography, mobile phase a: water (0.05% trifluoroacetic acid), mobile phase B: methanol; flow rate: 100 ml/min; elution was performed with 50% → 90% B phase in 30 minutes, detector UV 220/254 nm. Compound 40-4 was obtained (yellow oil, 320 mg, 33% yield). MS (ESI, m/z) 820.2/822.2[ 2 ], [ M + H ]] ⁺ 。

Step 4

Chiral resolution of compound 40-4 (320 mg) from step 3 by supercritical liquid chromatography: CHIRAL column CHIRAL ART amide-SA, 3 × 25 cm, 5 microns; mobile phase A: supercritical carbon dioxide, mobile phase B: isopropanol (0.5%, 2 mol/l ammonia methanol); flow rate: 60 ml/min; column temperature: 35 ℃; eluting with 35% mobile phase B; detector UV222 nm, resulting in two products. The product of shorter retention time (5.15 minutes) was compound 40-4a (white solid, 123 mg, recovery 38%), compound 40-4a ] ⁺ (ii) a The product of shorter retention time (6.32 min) was compound 40-4b (white solid, 106 mg, 33% yield), compound 40-4b] ⁺ 。

Step 5

To a reaction flask were added sequentially compound 40-4a (100 mg, 0.116 mmol, 1 eq.) and sodium periodate (130.43 mg, 0.580 mmol, 5 mm at 0 deg.C with stirringAmount), carbon tetrachloride (2.0 ml), acetonitrile (2 ml), to which was slowly added a solution of ruthenium trichloride (2.53 mg, 0.012 mmol, 0.1 eq) in water (3 ml). The mixture was reacted at 0 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. The crude product obtained was purified by reverse phase chromatography (C18 column), mobile phase a: water (0.1% ammonia water, ammonia content in ammonia water 25% -28%); mobile phase B, methanol, eluted with 30% → 95% of phase B within 20 minutes; detector UV254/220 nm; compound 40-5a was obtained (yellow solid, 60.0 mg, 59% yield). MS (ESI, m/z) 838.5/840.5[ 2 ], [ M + H ]] ⁺ 。

Step 6

To a solution of compound 40-5a (50.00 mg, 0.057 mmol, 1.00 eq) in N, N-dimethylformamide (1 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (27.23 mg, 0.068 mmol, 1.2 eq) with stirring at room temperature. The mixture was stirred at 25 ℃ for 10 minutes, and (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group was added to the reaction mixture ]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (30.65 mg, 0.063 mmol, 1.1 eq) and N, N-diisopropylethylamine (29.31 mg, 0.228 mmol, 4 eq). The mixture was stirred at 25 ℃ for a further 2 hours, the progress of the reaction being monitored by liquid chromatography and thin-layer chromatography. After the reaction was complete, the resulting mixture was purified by high performance liquid chromatography using Column XBridge Prep C18 OBD Column 30x50 mm, 5 μm; a mobile phase A: water (0.05% ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution was performed with 55% → 80% B phase over 8 minutes, detector UV 220 nm. Compound 40-6a was obtained (white solid, 30 mg, 40% yield). MS (ESI, m/z) 1250.0/1252.1[ m ] +H] ⁺ 。

Step 7

To a solution of compound 40-6a (30.00 mg, 0.114 mmol, 1.00 eq) in methanol (2 ml) was added dropwise a solution of 1, 4-dioxane (4 mol/l, 2 ml) of hydrochloric acid with stirring at 0 ℃. The mixture was allowed to react at room temperature for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. The crude product was purified by high performance liquid chromatography using a Column XBridge Shield RP18 OBD Column,19 × 150 mm, 5 μm; a mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was performed with 5% → 5% B in 1.5 minutes, 5% → 13% B in 2 minutes, and 13% → 39% B phase in 9 minutes, with detector UV 220 nm. Compound 40 was obtained (white solid, 14.2 mg, 51% yield). MS (ESI, m/z): 1106.1/1108.1[ m ] +H ] ⁺ 。 ¹ H NMR(300MHz,CD ₃ OD)δ8.89(s,1H),8.73–8.34(m,2H),7.76–7.61(m,2H),7.52–7.37(m,4H),7.34–7.19(m,2H),7.00(d,J＝2.6Hz,1H),4.79–4.62(m,3H),4.61–4.44(m,5H),4.44–4.33(m,1H),4.02–3.51(m,14H),3.25–3.01(m,2H),2.66–2.39(m,7H),2.37–2.16(m,2H),2.16–1.90(m,10H),1.09–0.95(m,9H),0.89–0.75(m,3H)； ¹⁹ F NMR(282MHz,CD ₃ OD)δ-117.19,-120.99,-124.04。

EXAMPLE 41

(2S, 4R) -1- ((S) -2- (4- ((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) propyl) (methyl) amino) butyrylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide dihydrochloride 41

Step 1

To a solution of compound 16-3a (300 mg, 0.465 mmol, 1.00 eq) and compound 4-1 (93.81 mg, 0.567 mmol, 1.2 eq) in anhydrous tetrahydrofuran (3 ml) was slowly added a solution of potassium tert-butoxide in tetrahydrofuran (1 mol per liter, 0.56 ml, 0.56 mmol, 1.2 eq) under nitrogen protection at 0 ℃. The resulting mixture was reacted at 0 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, 10 ml of water is added into the system to quench the reaction, the water phase is extracted by ethyl acetate (10 ml x 3), the organic phase is dried by anhydrous sodium sulfate after combination, the drying agent is removed by filtration, and the filtrate is concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, giving compound 41-1 (yellow solid, 231 mg, 64% yield). MS (ESI, m/z) 734.5/736.5[ 2 ] M + H ] ⁺ 。

Step 2

41-1 (160 mg, 0.207 mmol, 1.0 equiv.), sodium periodate (186.42 mg, 0.828 mmol, 4.0 equiv.), carbon tetrachloride (1.0 mL), acetonitrile (1 mL) were added to the flask sequentially with stirring at 0 deg.C. Then a solution of ruthenium trichloride (2.25 mg, 0.009 mmol, 0.05 eq) in water (1.5 ml) was slowly added thereto. The mixture was reacted at 0 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, 15 ml of water is added into the system to dilute the reaction solution, chloroform/isopropanol (3/1, 20 ml of x 3) is used for extraction, organic phases are combined, the organic phases are dried by anhydrous sodium sulfate, drying agents are removed through filtration, and the filtrate is concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) in 20 minutes with 5% → 95% acetonitrile/water mobile phase (0.1% ammonium bicarbonate)Line elution; detector UV254 nm; compound 41-2 was obtained (white solid, 70 mg, 40% yield). MS (ESI, m/z) 752.4/754.4[ M + H ]] ⁺ 。

Step 3

41-2 (70 mg, 0.088 mmol, 1.0 eq), 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (53.07 mg, 0.132 mmol, 1.5 eq) and N, N-dimethylformamide (2 ml) were added to the flask, in that order, with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 15 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group was added to the above reaction system ]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] carbonyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (41.37 mg, 0.088 mmol, 1 eq) and N, N-diisopropylethylamine (48.1 mg, 0.354 mmol, 4 eq). The mixture was allowed to react at 25 ℃ for a further 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, the crude product was purified by reverse phase flash chromatography (C18 column) eluting with 10% → 95% methanol/water mobile phase over 10 minutes; detector UV220 nm; compound 41-3 was obtained (yellow solid, 54 mg, 49% yield). MS (ESI, m/z) 1164.7/1166.7[ M + H ]] ⁺ 。

Step 4

To a solution of compound 41-3 (54 mg, 0.044 mmol, 1.00 eq) in methanol (2.0 ml) was added a solution of 1, 4-dioxane (4 mol per liter, 2.0 ml) of hydrochloric acid with stirring at 0 ℃. The resulting mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, the crude product is obtained by vacuum concentration, and the obtained crude product is purified by a reversed phase flash chromatography column (C18 column) at 30 minutesEluting with 5% → 95% methanol/water mobile phase (0.1% hydrochloric acid) in a short time; detector UV220/254 nm; compound 41 was obtained (yellow solid, 22.5 mg, 45% yield). MS (ESI, m/z) 1020.5/1022.5[ m + H ] ] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ10.71–10.52(m,1H),10.10–9.96(m,1H),9.88–9.68(m,1H),9.12(s,1H),8.72–8.60(m,1H),8.19–8.09(m,1H),8.06(d,J＝1.6Hz,1H),7.88(d,J＝8.3Hz,1H),7.55–7.41(m,5H),7.38(d,J＝2.4Hz,1H),7.34–7.23(m,2H),7.17(d,J＝2.4Hz,1H),4.67–4.38(m,8H),4.35–4.17(m,3H),4.09–3.93(m,2H),3.78–3.65(m,2H),3.40–3.02(m,4H),2.83(d,J＝4.7Hz,3H),2.52(s,3H),2.44–2.33(m,2H),2.32–2.21(m,2H),2.17–1.90(m,8H),1.01(s,9H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-121.99。

Example 42

(2S, 4R) -1- ((S) -2- ((1R, 4R or 1S, 4S) -4- (2- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) ethyl) cyclohexane-1-carboxamide-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide formate 42a; (2S, 4R) -1- ((S) -2- ((1S, 4S or 1R, 4R) -4- (2- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) ethyl) cyclohexane-1-carboxamide-3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide formate 42b

Step 1

To 2- (4- (ethoxycarbonyl) cyclohexyl) acetic acid (1.0 g, 4.4 mmol, 1.0 eq.) in anhydrous tetrahydrofuran (1 ml) was added with stirring at 0 deg.CTo the solution was slowly added dropwise a solution of borane in tetrahydrofuran (1 mol/l, 6.6 ml, 6.6 mmol, 1.0 eq). The mixture was stirred at 25 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction solution was slowly poured into ice water, extracted with ethyl acetate (30 ml × 3), and the combined organic phases were dried over anhydrous sodium sulfate. Filtering to remove insoluble substances, and concentrating the organic phase under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 9% methanol/dichloromethane and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and give compound 42-1 (yellow oil, 698 mg, 74% yield). MS (ESI, m/z) 201.1[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ4.19–4.06(m,2H),3.69(t,J＝6.6Hz,2H),2.28–2.15(m,1H),2.03–1.92(m,2H),1.88–1.78(m,2H),1.64–1.35(m,5H),1.34–1.21(m,3H),1.05–0.90(m,2H)。

Step 2

To a solution of compound 42-1 (690 mg, 3.2 mmol, 1.0 eq), N-dimethylaminopyridine (42 mg, 0.3 mmol, 0.1 eq) and triethylamine (1.44 ml, 9.8 mmol, 3.0 eq) in dichloromethane (6 ml) was slowly added p-toluenesulfonyl chloride (788 mg, mmol, 1.2 eq) with stirring at 0 degrees celsius under nitrogen protection. The resulting mixture was stirred at 20 ℃ for 2 hours. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was slowly poured into ice water, followed by extraction with dichloromethane (20 ml × 3), and the combined organic phases were dried over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the organic phase under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 50% methyl t-butyl ether/petroleum ether mobile phase gradient and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent and give compound 42-2 (pale yellow oil, 1.1 g, yield 80%). MS (ESI, m/z) 355.1[ 2 ], [ M ] +H] ⁺ 。 ¹ H NMR(400MHz,CDCl ₃ )δ7.79(d,J＝8.1Hz,2H),7.35(d,J＝8.0Hz,2H),4.15–4.02(m,4H),2.45(s,3H),2.20–2.10(m,1H),1.96–1.89(m,2H),1.73–1.64(m,2H),1.57–1.51(m,2H),1.41–1.28(m,3H),1.27–1.21(m,3H),0.95–0.82(m,2H)。

Step 3

To the reaction flask were added, under stirring at 25 ℃, compound 42-2 (979 mg, 2.6 mmol, 1.0 eq), L-prolinol (307 mg, 2.8 mmol, 1.1 eq), potassium carbonate (763 mg, 5.2 mmol, 2 eq) and acetonitrile (10 ml) in that order. The resulting mixture was reacted at 60 ℃ for 3 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is completed, insoluble substances are removed by filtration, and the filtrate is concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 9% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 42-3 (pale yellow oil, 600 mg, 76% yield). MS (ESI, m/z) 284.1[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.17–4.08(m,2H),3.80–3.69(m,1H),3.63–3.50(m,1H),3.47–3.31(m,1H),3.03–2.78(m,1H),2.57–2.36(m,2H),2.28–2.16(m,1H),2.06–1.73(m,9H),1.66–1.23(m,8H),1.09–0.89(m,2H)。

Step 4

To a solution of compound 16-3a (1.18 g, 1.92 mmol, 1.0 eq) and compound 42-3 (600 mg, 2.12 mmol, 1.1 eq) in anhydrous tetrahydrofuran (10 ml) was added dropwise a solution of potassium tert-butoxide in tetrahydrofuran (1 mol/l, 2.88 ml, 2.88 mmol, 1.5 eq) with stirring at-20 ℃ under nitrogen blanket. The resulting mixture was stirred at-20 ℃ for an additional 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction mixture was slowly poured into ice water, followed by ethyl acetate (30 ml. Times.3)) Extraction, combination and drying of the organic phase over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 9% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 42-4 (white solid, 791 mg, 48% yield). MS (ESI, m/z) 860.2/862.2[ M + H ]] ⁺ 。

Step 5

Chiral resolution of the compound 42-4 (576 mg) obtained in step 4 by preparative chiral high performance liquid chromatography: chiral column Lux5m Cellulose-2,3x25 cm, 5 micron; mobile phase A: supercritical carbon dioxide, mobile phase B: (ethanol: acetonitrile = 1; flow rate: 70 ml/min; elution was performed with 35% phase B over 25 minutes, detector UV 225 nm, to give two products. The product with a shorter retention time (8.68 min) was compound 42-4a (white solid, 49 mg, 8% recovery), compound 42-4a: MS (ESI, m/z) 860.2/862.2[ 2 ], [ M + H ] ] ⁺ (ii) a The product at longer retention time (11.33 min) was compound 42-4b (white solid, 268 mg, 46% recovery), compound 42-4b: MS (ESI, m/z) 860.2/862.2[ 2 ], [ M + H ]] ⁺ 。

Step 6

To a mixed solution of compound 42-4b (130 mg, 19.0 mmol, 1.0 equiv) in water/methanol/tetrahydrofuran (2/1/2, 5 ml) was added lithium hydroxide (36 mg, 1.4 mmol, 10.0 equiv) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction mixture was concentrated under reduced pressure to remove excess reagents. The mixture was diluted by adding 6 ml of water to the system, and then 1 mol/l of dilute hydrochloric acid was added dropwise to adjust the pH to 5. The aqueous phase was washed with dichloromethane (10 mm)L x 3), the combined organic phases are dried over anhydrous sodium sulfate and the drying agent is removed by filtration; and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 9% methanol/dichloromethane and the resulting fraction was concentrated under reduced pressure to remove the solvent and afford compound 42-5b (white solid, 51 mg, 40% yield). MS (ESI, m/z) 832.2/834.2[ M + H ]] ⁺ 。

Step 7

To a solution of compound 42-5b (50 mg, 0.05 mmol, 1.0 eq) in N, N-dimethylformamide (1.00 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethylurea hexafluorophosphate (27 mg, 0.06 mmol, 1.2 eq) with stirring at 25 ℃. The mixture was stirred at 25 ℃ for 30 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group was added to the reaction mixture ]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (33 mg, 0.06 mmol, 1.2 eq) and N, N-diisopropylethylamine (41 μ l, 0.17 mmol, 4 eq). The mixture was stirred at 25 ℃ for 1 hour. The course of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the mixture was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 70% acetonitrile/water mobile phase (0.05% trifluoroacetic acid) over 25 minutes; detector UV254 nm; compound 42-6b was obtained (white solid, 84 mg, 43% yield). MS (ESI, m/z) 1244.2/1246.2[ m + H ], [] ⁺ 。

Step 8

To a solution of compound 42-6b (80.0 mg, 0.06 mmol, 1 eq) in methanol (2.0 ml) was added a solution of 1, 4-dioxane hydrochloric acid (4 mol/l, 2.0 ml) with stirring at 0 ℃. What is neededThe mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the excess reagent was removed by concentration under reduced pressure, and the resulting mixture was purified by preparative high performance liquid chromatography on column Xselect CSH F-Phenyl OBD,19 × 250 mm, 5 μm; a mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was performed with 15% → 35% B phase over 8 minutes, detector UV 254/220 nm. Compound 42b was obtained (white solid, 15 mg, 21% yield). MS (ESI, m/z) 1100.1/1102.1[ m + H ] ] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ10.08(s,1H),9.50(s,1H),8.99(s,1H),8.62–8.51(m,1H),8.04–7.95(m,1H),7.82(d,J＝8.3Hz,1H),7.66(d,J＝9.2Hz,1H),7.49–7.36(m,5H),7.31(d,J＝2.4Hz,1H),7.25–7.16(m,2H),7.08(d,J＝2.4Hz,1H),5.23–5.08(m,1H),4.80–4.39(m,7H),4.39–4.18(m,3H),4.18–4.09(m,2H),3.94–3.79(m,2H),3.72–3.58(m,2H),3.12–2.87(m,1H),2.55(s,1H),2.45(s,3H),2.36–2.23(m,1H),2.22–2.10(m,1H),2.09–1.78(m,9H),1.77–1.62(m,4H),1.62–1.56(m,1H),1.56–1.42(m,2H),1.39–1.13(m,4H),1.05–0.70(m,11H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-121.90。

Following the procedure described in step 6, step 7, step 8, starting from compound 42-4a, compound 42a (white solid, 2.3 mg), MS (ESI, m/z): 1100.1/1102.1[ M ] +H ] +] ⁺ 。

Example 43

4- ((3- (3- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octyl-3-yl) -6-chloro-8-fluoro-7- (3-hydroxynaphthalen-1-yl) quinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) propoxy) propyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione Tritrifluoroacetate 43

Step 1

To a solution of N- (3-hydroxypropyl) carbamic acid tert-butyl ester (10 g, 54.215 mmol, 1.00 eq) and N, N-diisopropylethylamine (11.80 g, 86.744 mmol, 1.6 eq) in dichloromethane (100.0 ml) was slowly added methanesulfonyl chloride (8.50 g, 70.480 mmol, 1.3 eq) with stirring at 0 degrees celsius. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by thin layer chromatography. After completion of the reaction, the reaction was quenched by pouring the reaction solution into ice water and the aqueous phase was extracted with dichloromethane (200 ml × 3). After combination, the organic phase is dried by anhydrous sodium sulfate, the drying agent is removed by filtration, and the filtrate is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 50% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 43-1 (yellow oil, 14 g, 96% yield). ¹ H NMR(300MHz,CDCl ₃ )δ4.33–4.29(m,2H),3.30–3.26(m,2H),3.05(s,3H),2.00–1.91(m,2H),1.46(s,9H)。

Step 2:

to a solution of 1, 3-propanediol (26.64 g, 332.639 mmol, 6.57 eq) in N, N-dimethylformamide (253 ml) was added sodium hydride (60%, 4.05 g, 101.260 mmol, 2.00 eq) with stirring at 0 degrees celsius under nitrogen. After the mixture was stirred at 0 ℃ for 0.5 h, compound 43-1 (13.5 g, 50.630 mmol, 1.00 eq) was added. The reaction solution was reacted at 25 ℃ for 16 hours, and the progress of the reaction was monitored by thin layer chromatography. After the reaction was complete, the reaction solution was washed with saturated brine (1000 ml × 3), the organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give a crude product. Crude productPurification by silica gel column chromatography eluting with a 0% → 10% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 43-2 (yellow oil, 4.5 g, 36% yield). ¹ H NMR(400MHz,CDCl ₃ )δ3.78(t,J＝5.8Hz,2H)3.59(t,J＝5.8Hz,2H),3.50(t,J＝5.9Hz,2H),3.23(t,J＝6.5Hz,2H),1.86–1.79(m,2H),1.78–1.70(m,2H),1.44(s,9H)。

And 3, step 3:

to a solution of compound 43-2 (400 mg, 1.629 mmol, 1.00 eq) in methanol (2 ml) was added dropwise a solution of hydrochloric acid in 1, 4-dioxane (4 mol/l, 2 ml) with stirring at 25 ℃. The mixture was allowed to react for 2 hours at 25 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give crude product 43-3 (520 mg). The crude product was used directly in the next reaction. MS (ESI, m/z) 134.2[ m ] +H ] ⁺ 。

And 4, step 4:

to a mixture of compound 43-3 (297.99 mg, 2.125 mmol, 1.20 eq), N-methylpyrrolidone (5.0 ml) and 2- (2, 6 oxo-piperidin-3-yl) -4-fluoro-isoindole-1, 3-dione (515 mg, 1.771 mmol, 1.00 eq) was added dropwise N, N-diisopropylethylamine (722.90 mg, 5.313 mmol, 3.00 eq) at 25 degrees celsius with stirring under nitrogen. The resulting mixture was reacted at 90 ℃ for 2 hours, the course of the reaction being monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the reaction solution was cooled to 25 ℃. The mixture was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 60% acetonitrile/water mobile phase over 20 minutes; detector UV254 nm; a crude product of the compound is obtained. Purifying the crude product by silica gel column chromatographyElution with a 0% → 5% methanol/dichloromethane mobile phase gradient and the resulting fraction removed the solvent by rotary evaporation under reduced pressure to give compound 43-4 (yellow solid, 300 mg, 41% yield). MS (ESI, m/z) 390.1[ 2 ], M + H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ11.10(s,1H),7.64–7.49(m,1H),7.17–6.91(m,2H),6.80–6.54(m,1H),5.76(s,1H),5.15–4.93(m,1H),4.38(s,1H),3.49–3.34(m,8H),2.96–2.79(m,1H),2.64–2.57(m,1H),2.11–1.96(m,1H),1.86–1.74(m,2H),1.73–1.59(m,2H)。

And 5:

to a solution of compound 43-4 (300 mg, 0.732 mmol, 1.00 eq) in dichloromethane (6 ml) was added 1, 1-triacetoxy-1, 1-dihydro-1, 2-benziodo-3 (1H) -one (392.11 mg, 0.878 mmol, 1.20 eq) with stirring at 0 degrees celsius under nitrogen protection. The resulting mixture was reacted at 25 ℃ for 2 hours, the course of which was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 5% methanol/dichloromethane mobile phase gradient and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compound 43-5 (yellow solid, 130 mg, 43% yield). MS (ESI, m/z) 388.2[ m ] +H ] ⁺ 。

Step 6

To a solution of compound 35-2 (240 mg, 0.33 mmol, 1 eq) in methanol (3.0 ml) was added a solution of 1, 4-dioxane (4 mol/l, 3.0 ml) of hydrochloric acid with stirring at 0 ℃. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, concentration was carried out under reduced pressure to obtain crude product 43-6 (310 mg). MS (ESI, m/z): 574.2/576.2[ m ] +H] ⁺ 。

Step 7

To a solution of compound 43-6 (300 mg, 0.318 mmol, 1.00 eq) and N, N-diisopropylethylamine (151.28 mg, 1.113 mmol, 3.50 eq) in dichloromethane (6.0 ml) was added di-tert-butyl dicarbonate (72.99 mg, 0.318 mmol, 1.00 eq) with stirring at 0 ℃. The resulting mixture was stirred at 0 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 6% methanol/dichloromethane mobile phase gradient, and the resulting fraction was subjected to rotary evaporation under reduced pressure to remove the solvent, whereby compound 43-7 (yellow solid, 210 mg) was obtained. MS (ESI, m/z) 674.3/676.3[ M ] +H ] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ10.02(s,1H),8.00(d,J＝1.6Hz,1H),7.81(d,J＝8.3Hz,1H),7.49–7.37(m,1H),7.29(d,J＝2.4Hz,1H),7.25–7.18(m,2H),7.07(d,J＝2.4Hz,1H),5.93–5.79(m,1H),5.16(d,J＝16.9Hz,1H),5.03(d,J＝10.3Hz,1H),4.50–4.33(m,3H),4.31–4.20(m,2H),4.21–4.09(m,1H),3.65–3.49(m,3H),3.04–2.91(m,2H),2.90–2.79(m,1H),2.31–2.16(m,1H),1.98–1.59(m,8H),1.47(s,9H)。

Step 8

To a three-necked flask were added sequentially compounds 43-7 (200 mg, 0.282 mmol, 1.00 eq), 1, 3-dimethylbarbituric acid (69.48 mg, 0.423 mmol, 1.50 eq), tetrakis (triphenylphosphine) palladium (17.14 mg, 0.014 mmol, 0.05 eq) and dichloromethane (6 ml) under nitrogen protection at 25 ℃. The resulting mixture was reacted at 25 ℃ for 5 hours, the course of the reaction being monitored by chromatography on liquid and thin layer chromatography. After the reaction is finished, the excessive solvent is removed by decompression and concentration to obtain a crude product. Crude productPurification was performed by reverse phase flash chromatography (C18 column) eluting with 20% → 60% acetonitrile/water mobile phase (0.1% formic acid) over 20 minutes, detector UV254 nm to give compound 43-8 (light yellow solid, 120 mg, 63% yield). MS (ESI, m/z) 634.2/636.2[ 2 ], [ M + H ]] ⁺ 。

Step 9

To a solution of compound 43-8 (120 mg, 0.180 mmol, 1.00 eq), compound 43-5 (109.96 mg, 0.270 mmol, 1.50 eq) and acetic acid (22.73 mg, 0.360 mmol, 2.00 eq) in methanol (7 ml) was added sodium cyanoborohydride (17.84 mg, 0.270 mmol, 1.50 eq) with stirring at 25 ℃. The resulting mixture was reacted at 25 ℃ for 1 hour, and the course of the reaction was monitored by chromatography on liquid and thin layer chromatography. After the reaction is finished, the reaction liquid is decompressed and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 5% methanol/dichloromethane and the resulting fraction was rotary evaporated under reduced pressure to remove the solvent and afford compounds 43-9 (yellow solid, 75 mg, 37% yield). MS (ESI, m/z) 1005.4/1007.4[ 2 ], [ M ] +H ] ⁺ 。

Step 10

To a solution of compound 43-9 (70 mg, 0.069 mmol, 1.00 eq) in dichloromethane (4.0 ml) was added trifluoroacetic acid (1 ml) with stirring at room temperature. The resulting mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction liquid is decompressed and concentrated to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 5% → 50% acetonitrile/water mobile phase (0.1% trifluoroacetic acid) over 10 minutes with detector UV254 nm to give compound 43 (yellow-green solid, 40 mg, 43% yield)。MS(ESI,m/z):905.5/907.5[M+H] ⁺ ； ¹ H NMR(300MHz,DMSO-d ₆ )δ7.92(d,J＝14.9Hz,1H),7.81(d,J＝8.3Hz,1H),7.53–7.42(m,2H),7.31(d,J＝2.4Hz,1H),7.27–7.13(m,2H),7.10–7.06(m,1H),6.98–6.91(m,2H),5.10–4.98(m,1H),4.70–4.40(m,4H),4.25–4.12(m,2H),4.00–3.90(m,1H),3.88–3.71(m,2H),3.69–3.56(m,2H),3.47–3.37(m,4H),3.32–3.15(m,4H),2.94–2.78(m,1H),2.67–2.54(m,2H),2.33–2.20(m,1H),2.08–1.88(m,10H),1.81–1.68(m,2H)； ¹⁹ F NMR(282MHz,DMSO-d ₆ )δ-74.13,-121.86,-121.92。

Example 44

(2S, 4R) -1- ((2S) -2- (3- (3- ((2R, 7aS) -7a- ((4- (3, 8-diazabicyclo [3.2.1] octan-3-yl) -7- (8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl) -8-fluoropyridine [4,3-d ] pyrimidin-2-yl) oxy) methyl) hexahydro-1H-pyrrolizin-2-yl) oxy) propoxy) propionamido) -3, 3-dimethylbutyryl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide trifluoroacetate (44)

Step 1:

to a solution of compound 2, 5-dioxo-1H-tetrahydropyrrolazine-7 a (5H) -carboxylic acid ethyl ester (10.5 g, 47.226 mmol, 1 eq) in anhydrous tetrahydrofuran (150 ml) was added lithium aluminum hydride (9.43 g, 236.13 mmol, 5 eq) portionwise with stirring at 0 degrees celsius under nitrogen protection. The mixture was allowed to react for 16 hours at 60 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was completed, the mixture system was cooled to room temperature, and water (9.5 ml), a 15% aqueous solution of sodium hydroxide (9.5 ml) and water (28.5 ml) were added in this order. Filtering to remove insoluble substances, and concentrating the filtrate under reduced pressure to obtain crude product. Purifying the obtained crude product by high performance liquid chromatography, wherein the column XB is a chromatographic column ridge Prep Phenyl OBD Column,19 × 150 mm, 5 μm; mobile phase A: water (0.05% ammonia), mobile phase B: methanol; flow rate: 25 ml/min; elution was performed with 3% → 30% B phase in 7 minutes, detector UV200/220 nm. Compound 44-1 was obtained (pale yellow oily liquid, 2.66 g, 34% yield). MS (ESI, m/z) 158.2[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ4.54(s,2H),4.47–4.35(m,1H),3.30–3.13(m,3H),2.99–2.78(m,2H),2.61–2.52(m,1H),2.25–2.16(m,1H),2.04–1.86(m,1H),1.83–1.55(m,4H)。

Step 2:

to a solution of compound 44-1 (2.6 g, 15.711 mmol, 1 eq) in N, N-dimethylformamide (30 ml) was added tert-butyldiphenylchlorosilane (4.91 g, 16.97 mmol, 1.08 eq) in portions with stirring at-40 ℃ under nitrogen protection. The mixture was allowed to react for 5 hours at 25 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the mixture system was purified by reverse phase chromatography (C18 column) to obtain a crude product, which was eluted with 5% → 95% methanol/water mobile phase (0.1% formic acid) within 25 minutes; detector, UV254/220 nm. The crude product is further purified by preparative supercritical liquid chromatography under the following conditions: chiral column YMC-PACK CN,3x25 cm, 5 micron; mobile phase A: supercritical carbon dioxide fluid, mobile phase B: methanol (0.1-2 mol/l ammonia methanol); flow rate: 180 ml/min; elution with 30% phase B over 12 min, detector UV 220/204 nm gave compound 44-2 (pale yellow oily liquid, 2.4 g, 36% yield). MS (ESI, m/z) 396.1[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ9.14–9.06(m,4H),8.88–8.77(m,6H),5.89–5.80(m,1H),4.83–4.75(m,2H),4.72–4.65(m,1H),4.52–4.43(m,1H),4.30–4.21(m,1H),4.09–4.00(m,1H),3.80–3.72(m,1H),3.40–3.30(m,2H),3.21–3.13(m,2H),3.08–3.02(m,1H),2.50(s,9H)。

And 3, step 3:

chiral resolution of compound 44-2 (2.4 g) from step 2 by preparative supercritical liquid chromatography: chiral column Lux 5m Cellulose-4,3x25 cm, 5 micron; a mobile phase A: supercritical carbon dioxide fluid, mobile phase B: methanol (0.1% 2 mol/l ammonia methanol); flow rate: 20 ml/min; elution was performed with 35% phase B over 12 minutes, detector: UV 204 nm, resulting in two products. The product with shorter retention time (5.83 min) is compound 44-3 (yellow oil, 1.05 g, recovery 44%), MS (ESI, m/z) 396.1[ M ] +H] ⁺ ；[α] ²⁵ _D ＝+11.6(c＝0.33g/100mL,MeOH)。

And 4, step 4:

to a solution of compound 44-3 (800 mg, 1.921 mmol, 1.0 eq) and sodium iodide (333.42 mg, 2.113 mmol, 1.1 eq) in anhydrous N, N-dimethylformamide (8 ml) was added dropwise a solution of 1 mol/l potassium tert-butoxide in tetrahydrofuran (3.84 ml, 3.84 mmol, 2.0 eq) at 0 ℃ with stirring under nitrogen protection. The mixture was allowed to react for 16 hours at 25 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was purified by a reverse phase chromatography column (C18 column) to obtain a crude product, which was eluted with a methanol/water mobile phase (0.1% formic acid) of 10% → 85% in 25 minutes; a detector: UV254/220 nm. Further purifying the obtained crude product by high performance liquid chromatography with chromatographic column YMC-Actus Triart C18 ExRS,30x150 mm, 5 μm; a mobile phase A: water (10 mmol/l ammonium bicarbonate), mobile phase B: methanol; flow rate: 60 ml/min; elution was performed with 85% → 95% B phase in 10 minutes, detector: UV 220/254 nm. Compound 44-4 was obtained (yellow oily liquid, 380 mg, 37% yield). MS (ESI, m/z) 508.3[ m + H ] ] ⁺ 。

And 5:

to a solution of compound 44-4 (350 mg, 0.655 mmol, 1.0 eq) in tetrahydrofuran (4 ml) was added dropwise a 1 mol/l solution of tetrabutylammonium fluoride in tetrahydrofuran (1.64 ml, 1.64 mmol, 2.5 eq) with stirring at 25 ℃. The mixture was allowed to react for 18 hours at 40 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, decompressing and concentrating to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a gradient of 0% → 10% methanol (1%, 7 mol/l methanolic ammonia)/dichloromethane and the resulting fraction was rotary distilled under reduced pressure to remove the solvent to give compound 44-5 (yellow oily liquid, 140 mg, 78% yield). MS (ESI, m/z) 270.2[ 2 ], [ M ] +H] ⁺ ， ¹ H NMR(400MHz,CDCl ₃ )δ5.87–5.77(m,1H),5.15–5.00(m,2H),4.05–4.00(m,1H),3.50–3.43(m,6H),3.25(s,2H),3.16–3.12(m,1H),3.10–3.04(m,1H),2.90–2.85(m,1H),2.82–2.78(m,1H),2.35–2.30(m,2H),2.10–2.05(m,1H),1.99–1.88(m,1H),1.84–1.72(m,6H)。

Step 6:

to a solution of compound 44-5 (110 mg, 0.388 mmol, 1.0 eq) and compound 38-6 (210 mg, 0.166 mmol, 1.2 eq) in tetrahydrofuran (3 ml) was added dropwise a solution of potassium tert-butoxide in tetrahydrofuran (0.58 ml, 0.58 mmol, 1.5 eq) at-10 ℃ with stirring under nitrogen protection. The mixture was allowed to react for 2 hours at-10 ℃ with stirring, the course of the reaction being monitored by liquid chromatography and thin layer chromatography. After the reaction, water (25 ml) was added to the reaction solution to quench the reaction, the resulting mixture was extracted with ethyl acetate (25 ml × 3), and the organic phases were combined; the organic phase was washed with saturated brine (30 ml. Times.1) and then washed with brine Drying with sodium sulfate, filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a gradient of 0% → 8% methanol/dichloromethane and the fractions were stripped of solvent by rotary evaporation under reduced pressure to give compound 44-6 (yellow solid, 190 mg, 68% yield). MS (ESI, m/z) 661.2/663.2[ M + H ]] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ8.73(s,1H),5.86–5.76(m,1H),5.15–5.01(m,2H),4.65–4.29(m,6H),4.20(s,1H),3.65(d,J＝7.5Hz,2H),3.55–3.43(m,7H),3.05(s,2H),2.35–2.29(m,3H),2.18(s,1H),2.11–1.91(m,6H),1.86–1.80(m,2H),1.78–1.65(m,3H),1.52(s,9H)。

And 7:

to a reaction flask were added, in order, compounds 44-6 (110 mg, 0.158 mmol, 1.0 equivalent) ((2-fluoro-6- (methoxymethoxy) -8- (4, 5-tetramethyl-1, 3, 2-dioxabenzaldehyde-2-yl) naphthalen-1-yl) ethynyl) triisopropylsilane (prepared with reference to WO2021041671, 110.8 mg, 0.205 mmol, 1.3 equivalents), tris (dibenzylideneacetone) dipalladium (15.2 mg, 0.016 mmol, 0.1 equivalent), n-butyl bis (1-adamantyl) phosphine (11.9 mg, 0.032 mmol, 0.2 equivalent), toluene (2 ml) and water (0.4 ml) at 25 degrees celsius with stirring under nitrogen protection. The mixture was reacted for 8 hours at 80 ℃ with stirring, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction mixture was cooled to room temperature, water (10 ml) was added to the reaction mixture to dilute the reaction mixture, the resulting mixture was extracted with ethyl acetate (10 ml × 3), and the organic phases were combined; the organic phase was washed with 30 ml of saturated brine, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography on silica gel eluting with a gradient of 0% → 8% methanol/dichloromethane and the fractions were stripped of solvent by rotary evaporation under reduced pressure to give compound 44-7 (yellow solid, 120 mg, 71% yield). MS (ESI, m/z) 1011.5[ 2 ] M + H ] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ9.06(s,1H),7.80–7.76(m,1H),7.51(d,J＝2.6Hz,1H),7.34–7.28(m,2H),5.8–5.76(m,1H),5.30(s,2H),5.14–5.02(m,2H),4.83–4.68(m,2H),4.54–3.99(m,6H),3.96–3.71(m,2H),3.61–3.45(m,11H),2.51–2.12(m,7H),2.10–1.93(m,5H),1.90–1.80(m,3H),1.52(s,9H),0.88–0.84(m,18H),0.63–0.46(m,3H)。

And 8:

to a mixed solution of compounds 44-7 (110 mg, 0.103 mmol, 1.0 equiv), ruthenium trichloride monohydrate (2.5 mg, 0.01 mmol, 0.1 equiv), water (3 ml), carbon tetrachloride (2 ml) and acetonitrile (2 ml) was added sodium periodate (116.3 mg, 0.515 mmol, 5.0 equiv) in portions under stirring at 0 ℃. The mixture was reacted for 1 hour at 0 ℃ with stirring, and the reaction was monitored by liquid mass. After the reaction was completed, water (10 ml) was added to the reaction solution, the resulting mixture was extracted with chloroform/isopropanol (3/1, 10 ml × 3), the organic phases were combined, dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by reverse phase chromatography (C18 column) eluting with 5% → 95% methanol/water mobile phase (0.1% ammonia) over 15 minutes; a detector: UV254/220 nm; compound 44-8 was obtained (white solid, 70 mg, 62% yield). MS (ESI, m/z) 1029.5[ m + H ], [ solution of calcium ] C] ⁺ 。

And step 9:

to a solution of compound 44-8 (70 mg, 0.065 mmol, 1.0 eq) in N, N-dimethylformamide (2 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (31.0 mg, 0.078 mmol, 1.2 eq) with stirring at 25 ℃. The mixture was reacted at 25 ℃ for 15 minutes. Then, (2S, 4R) -1- [ (2S) -2-amino group was added to the reaction solution -3, 3-dimethylbutyryl group]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (38.1 mg, 0.078 mmol, 1.2 equivalents) and N, N-diisopropylethylamine (35.1 mg, 0.26 mmol, 4 equivalents). The reaction solution was reacted at 25 ℃ for 1 hour, and the reaction process was monitored by liquid mass. After the reaction was completed, the mixture was purified by a reverse phase chromatography column (C18 column) eluting with 5% → 95% methanol/water mobile phase (0.1% trifluoroacetic acid) over 15 minutes; detector UV254/220 nm; compound 44-9 was obtained (yellow solid, 68 mg, 69% yield). MS (ESI, m/z) 721.7[ m/2 ] +H] ⁺ 。

Step 10:

to a solution of compound 44-9 (68 mg, 0.047 mmol, 1.0 eq) in acetonitrile (2.5 ml) was added 4 mol per liter of 1, 4-dioxane hydrochloride solution (0.5 ml) with stirring at 0 ℃. The mixture was reacted for 2 hours at 0 ℃ with stirring, the course of the reaction being monitored by liquid mass. After the reaction was completed, the solvent was removed by concentration under reduced pressure. To the mixture was added saturated sodium bicarbonate solution (10 ml), the resulting mixture was extracted with chloroform/isopropanol (3/1, 10 ml x 3), the organic phases were combined, dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the filtrate was concentrated under reduced pressure to give crude compound 44-10 (yellow solid, 60 mg, 95% yield). MS (ESI, m/z) 1297.6[ m ] +H ] ⁺ 。

Step 11:

cesium fluoride (76 mg, 0.46 mmol, 10 equiv.) is added to a solution of compound 44-10 (60 mg, 0.046 mmol, 1.0 equiv.) in N, N-dimethylformamide (2 ml) with stirring at 25 ℃. The mixture was reacted for 4 hours at 25 ℃ with stirring, the course of the reaction being monitored by liquid mass. After the reaction is finishedAnd filtering to remove excessive cesium fluoride, and concentrating the filtrate under reduced pressure to obtain a crude product. Purifying the obtained crude product by high performance liquid chromatography with Column XBridge Prep OBD C18 Column of 30x150 mm, 5 μm; mobile phase A: water (10 mmol/l ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution was carried out with 27% → 50% B phase in 7 minutes, detector UV 220/254 nm. Compound 44 was obtained (yellow solid, 30 mg, 49% yield). MS (ESI, m/z) 571.9[ m/2 ] +H] ⁺ ，1141.6[M+H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ10.20(s,1H),9.08(s,1H),8.98(s,1H),8.62–8.59(m,1H),8.00–7.90(m,2H),7.54–7.35(m,6H),7.17(d,J＝2.5Hz,1H),5.22–5.05(m,1H),4.62–4.52(m,2H),4.46–4.34(m,4H),4.29–4.01(m,4H),3.97–3.85(m,3H),3.78–3.47(m,6H),3.44–3.37(m,4H),3.21–2.99(m,2H),2.92–2.69(m,2H),2.58–2.53(m,1H),2.43(s,3H),2.38–2.26(m,1H),2.19–1.99(m,2H),1.98–1.61(m,12H),0.92(s,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-73.43,-110.72,-139.93。

Example 45

(2S, 4R) -1- ((S) -2- (3- (3- ((S) -2- ((((R or S) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl) -6, 8-difluoroquinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) propoxy) propylamino) -3, 3-dimethylbutyryl-4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide monoformate 45

Step 1

To a solution of compound 40-5a (65.00 mg, 0.074 mmol, 1.00 eq) in N, N-dimethylformamide (1.5 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (35.39 mg) with stirring at room temperature0.089 mmol, 1.2 equivalents). The mixture was stirred at 25 ℃ for 10 minutes, and (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) was added to the reaction mixture]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] amide]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (41.38 mg, 0.089 mmol, 1.2 eq) and N, N-diisopropylethylamine (40.10 mg, 0.296 mmol, 4 eq). The mixture was stirred at 25 ℃ for a further 2 hours, the progress of the reaction being monitored by liquid chromatography and thin-layer chromatography. After the reaction is completed, the obtained mixture is purified by high performance liquid chromatography, and a chromatographic Column Kinetex EVO C18 Column 21.2x150 mm is 5 microns; mobile phase A: water (0.05% ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 25 ml/min; elution was carried out with 80% → 95% B phase in 8 minutes, detector UV 220 nm. Compound 45-1 was obtained (white solid, 42 mg, 42% yield). MS (ESI, m/z) 1264.6[ m + H ], [ M ], [ solution of calcium ] and [ sic ], [ solution of calcium ] in (ESI, m/z) ] ⁺ 。

Step 2

To a solution of compound 45-1 (42.00 mg, 0.032 mmol, 1.00 eq) in methanol (1 ml) was added dropwise a solution of 1, 4-dioxane (4 mol/l, 1 ml) of hydrochloric acid with stirring at 0 ℃. The mixture was allowed to react at room temperature for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Purifying the crude product by high performance liquid chromatography with a chromatographic Column Sunfire prep C18 Column,30x150 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution was carried out with 18% → 35% B phase in 7 minutes, detector UV 220 nm. Compound 45 was obtained (white solid, 25.8 mg, 67% yield). MS (ESI, m/z) 1120.4[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ8.98(s,1H),8.38(d,J＝7.8Hz,1H),8.29(s,1H),7.89–7.74(m,2H),7.73–7.63(m,1H),7.49–7.29(m,6H),7.02(d,J＝2.6Hz,1H),5.01–4.81(m,1H),4.55–4.47(m,1H),4.47–4.37(m,1H),4.37–4.19(m,4H),4.12–4.02(m,1H),3.69–3.48(m,10H),3.43–3.30(m,3H),3.09–2.98(m,1H),2.93–2.76(m,2H),2.45(s,3H),2.40–2.15(m,4H),2.07–1.96(m,1H),1.95–1.86(m,1H),1.83–1.59(m,10H),1.36(d,J＝7.0Hz,3H),0.89(s,9H),0.79–0.65(m,3H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-118.286,-118.294,-119.111,-123.736,-123.742。

Example 46

4- ((3- (3- ((S) -2- ((((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl) -6, 8-difluoroquinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) propoxy) propyl) amino) -2- (2, 6-dioxopiperidin-3-yl) isoindoline-1, 3-dione dihydrochloride 46

Step 1

To compound 40-1 (650 mg, 1.261 mmol, 1.0 equiv.), compound 40-3 (525.91 mg, 1.387 mmol, 1.1 equiv.), 3- (tert-butyl) -4- (2, 6-dimethoxyphenyl) -2, 3-dihydrobenzo [ D ] was added under stirring at 25 ℃ under nitrogen protection ][1,3]To a mixed solution of oxygen, phosphine-pentyl-yoke (87.69 mg, 0.252 mmol, 0.2 eq) and tris (dibenzylideneacetone) dipalladium (121.54 mg, 0.126 mmol, 0.1 eq) in toluene/water (5.5 ml/1.1 ml) was added potassium phosphate (563.44 mg, 2.522 mmol, 2 eq) in portions. The mixture was reacted at 80 ℃ for 4 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature, and the mixture was concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a 0% → 20% ethyl acetate/petroleum ether mobile phase gradient, and the resulting fraction was concentrated under reduced pressure to remove the solvent and give compound 46-1 (white solid, 550 mg, 64% yield). MS (ESI, m/z) 643.2/644.2[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.74–7.69(m,1H),7.56(d,J＝2.7Hz,1H),7.45–7.41(m,1H),7.32–7.26(m,1H),7.11(d,J＝2.7Hz,1H),5.31(s,2H),4.52–4.41(m,4H),3.71–3.65(m,2H),3.55(s,3H),2.67–2.54(m,1H),2.47–2.33(m,1H),2.06–1.96(m,2H),1.91–1.76(m,2H),1.55(s,9H),0.88–0.83(m,3H)。

Step 2

Compound 35-1 (118.36 g, 0.796 mmol, 1.1 eq) was added to a solution of compound 46-1 (490 mg, 0.724 mmol, 1 eq), cesium carbonate (496.51 mg, 1.448 mmol, 2 eq) and triethylene diamine (17.09 mg, 0.145 mmol, 0.2 eq) in N, N-dimethylformamide (5 ml) at 25 degrees c with stirring under nitrogen. The mixture was stirred at 90 ℃ for 2 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, the reaction mixture was diluted with 200 ml of water, the aqueous layer was extracted with ethyl acetate (300 ml × 3), and the combined organic layers were washed with saturated brine (100 ml × 3) and dried over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was concentrated under reduced pressure to remove the solvent, yielding compound 46-2 (white solid, 360 mg, 63% yield). MS (ESI, m/z) 748.4[ m ] +H ] ⁺ 。

Step 3

Chiral resolution of compound 46-2 (360 mg) from step 2 by supercritical liquid chromatography: CHIRAL column chiralart cell-SC, 3 × 25 cm, 5 μm; mobile phase A: supercritical carbon dioxide, mobile phase B: ethanol (0.5%, 2 mol/l ammonia methanol); flow rate: 60 ml/min; column temperature: 35 ℃ is carried out; eluting with 35% mobile phase B; detector UV222 nm to obtain two products. The product, with shorter retention time (4.85 minutes), is compound 46-2a (white solid, 152 mg, recovery 42%), MS (ESI, m/z) 748.4[ 2 [ M + H ])] ⁺ (ii) a The product of longer retention time (5.16 minutes) is compound 46-2b (white solid, 153 mg, 42% yield), MS (ESI, m/z) 748.4[ 2 [ M + H ])] ⁺ 。

Step 4

Compound 46-2a (60 mg, 0.08 mmol, 1.0 eq), 1, 3-dimethylbarbituric acid (18.79 mg, 0.12 mmol, 1.5 eq), tetrakis (triphenylphosphine) palladium (4.64 mg, 0.004 mmol, 0.05 eq) and dichloromethane (0.8 ml) were added sequentially to a 50 ml three-necked flask under nitrogen protection at 25 ℃. The resulting mixture was reacted at 0 ℃ for 2 hours, the course of the reaction was monitored by chromatography on liquid and thin layer chromatography. After the reaction is finished, concentrating under reduced pressure to remove the redundant solvent to obtain a crude product. The crude product was purified by reverse phase flash chromatography (C18 column) eluting with 10% → 50% acetonitrile/water mobile phase (0.1% formic acid) over 20 minutes, detector UV254 nm to give compound 46-3a (yellow solid, 50 mg, 70% yield). MS (ESI, m/z) 708.3[ 2 ], [ M ] +H ] ⁺ 。

Step 5

To a solution of compound 46-3a (44 mg, 0.058 mmol, 1.00 eq), compound 43-5 (27.13 mg, 0.07 mmol, 1.20 eq) and acetic acid (5.26 mg, 0.087 mmol, 1.5 eq) in methanol (0.5 ml) was added sodium cyanoborohydride (11 mg, 0.174 mmol, 3.0 eq) with stirring at 25 ℃. The resulting mixture was reacted at 25 ℃ for 1 hour, and the course of the reaction was monitored by chromatography on liquid and thin layer chromatography. After the reaction is finished, the reaction liquid is decompressed and concentrated to obtain a crude product. Purifying the crude product by silica gel column chromatographyElution with a 0% → 5% methanol/dichloromethane mobile phase gradient gave a fraction which was rotary evaporated under reduced pressure to remove the solvent and afford compound 46-4a (white solid, 38 mg, 56% yield). MS (ESI, m/z) 1079.5[ 2 ], [ M ] +H] ⁺ 。

Step 6

To a solution of compound 46-4a (38 mg, 0.033 mmol, 1.00 eq) in methanol (2 ml) was added dropwise a solution of 1, 4-dioxane (4 mol/l, 2 ml) of hydrochloric acid, with stirring at 0 ℃. The mixture was allowed to react at room temperature for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Purifying the crude product by high performance liquid chromatography with chromatographic column YMC-actual Triart C18 of 30x150 mm and 5 μm; mobile phase A: water (0.05% hydrochloric acid), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution was performed with 5% → 5% B in 1.5 minutes and 20% → 45% B phase in 7 minutes, detector UV 220 nm. Compound 46 was obtained (yellow solid, 19.7 mg, 58% yield). MS (ESI, m/z) 935.4[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(400MHz,DMSO-d ₆ )δ11.10(s,1H),10.45(s,1H),10.12(s,1H),9.73(s,1H),9.47(s,1H),7.80–7.67(m,2H),7.53–7.50(m,1H),7.41–7.32(m,2H),7.08–6.97(m,2H),6.94–6.91(m,1H),6.62(s,1H),5.12–5.02(m,1H),4.80–4.73(m,1H),4.69–4.63(m,1H),4.43–4.35(m,2H),4.16(s,2H),3.92(s,1H),3.84–3.76(m,2H),3.70–3.54(m,3H),3.50–3.46(m,3H),3.32–3.25(m,2H),3.22–3.10(m,2H),2.96–2.82(m,1H),2.60–2.55(m,1H),2.47–2.39(m,1H),2.33–2.21(m,3H),2.09–1.86(m,10H),1.81–1.70(m,2H),0.74–0.70(m,3H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-116.31,-119.15,-123.10。

Example 47

(2S, 4R) -1- ((S) -2- (3- (3- ((S) -2- (((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -7- (8-ethynyl-7-fluoro-3-hydroxynaphthalen-1-yl) -6, 8-difluoroquinazolin-2-yl) oxy) methyl) pyrrolidin-1-yl) propoxy) propylamino) -3, 3-dimethylbutanoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) phenyl) pyrrolidine-2-carboxamide monoformate 47

Step 1

To compound 40-2 (800 mg, 1.14 mmol, 1.0 eq), compound 44-7 (676.61 mg, 1.254 mmol, 1.1 eq), 3- (tert-butyl) -4- (2, 6-dimethoxyphenyl) -2, 3-dihydrobenzo [ D ] was added under stirring at 25 deg.C under nitrogen protection][1,3]To a mixed solution of oxygen, phosphine-pentalene (76.87 mg, 0.228 mmol, 0.2 eq) and tris (dibenzylideneacetone) dipalladium (106.53 mg, 0.114 mmol, 0.1 eq) in toluene/water (8 ml/1.6 ml) was added potassium phosphate (493.88 mg, 2.280 mmol, 2 eq) in portions. The mixture was reacted at 80 ℃ for 2 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to room temperature, and the mixture was concentrated to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 8% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 47-1 (yellow semisolid, 720 mg, 65% yield). MS (ESI, m/z) 972.5[ m + H ] ] ⁺ 。

Step 2

Chiral resolution of compound 47-1 (720 mg) from step 3 by supercritical liquid chromatography: chiral column CHIRALPAK AD-H,3 × 25 cm, 5 microns; a mobile phase A: supercritical carbon dioxide, mobile phase B: isopropanol (0.5%, 2 mol/l ammonia methanol); flow rate: 50 ml/min; column temperature: 35 ℃ is carried out; elution with 30% mobile phase B(ii) a Detector UV222 nm, resulting in two products. The product with shorter retention time (2.00 min) is compound 47-1a (yellow solid, 310 mg, 43% recovery), MS (ESI, m/z): 972.5[ m ] +H] ⁺ (ii) a The product of shorter retention time (2.4 minutes) is compound 47-1b (yellow solid, 300 mg, 41% yield), MS (ESI, m/z): 972.5[ M ] +H] ⁺ 。

Step 3

To a reaction flask were added sequentially the compound 47-1a (280 mg, 0.274 mmol, 1 eq), sodium periodate (298.56 mg, 1.370 mmol, 5 eq), carbon tetrachloride (1.5 ml), acetonitrile (1.5 ml) at 0 ℃ with stirring, and a solution of ruthenium trichloride (6.29 mg, 0.027 mmol, 0.1 eq) in water (2.25 ml) was slowly added dropwise. The mixture was reacted at 0 ℃ for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. The crude product obtained was purified by reverse phase chromatography (C18 column), mobile phase a: water (0.1% ammonium bicarbonate); mobile phase B, methanol, eluting with 5% → 95% of phase B in 30 minutes; detector UV254/220 nm; compound 47-2a was obtained (white solid, 90 mg, 33% yield). MS (ESI, m/z) 990.5[ m ] +H ] ⁺ 。

Step 4

To a solution of compound 47-2a (80 mg, 0.077 mmol, 1.00 eq) in N, N-dimethylformamide (1 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (35.73 mg, 0.092 mmol, 1.2 eq) with stirring at room temperature. The mixture was stirred at 25 ℃ for 10 minutes, and (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group was added to the reaction mixture]-4-hydroxy-N- [ [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Methyl radical]Pyrrolidine-2-carboxamide hydrochloride (43.89 mg, 0.092 mmol, 1.2 equivalents) and N, N-diisopropylethylamine (40.08 mg, 0.308 mmol, 4 equivalents). The mixture was stirred at 25 ℃ for a further 2 hours, the progress of the reaction being monitored by liquid chromatography and thin-layer chromatography. After completion of the reaction, the resulting mixture was purified by preparative thin layer chromatography eluting with a 0% → 8% methanol/dichloromethane mobile phase gradient to give compound 47-3a (white solid, 80 mg, 74% yield). MS (ESI, m/z) 1402.7[ m ] +H] ⁺ 。

Step 5

Cesium fluoride (42.23 mg, 0.27 mmol, 5.00 equiv) was added to a solution of compound 47-3a (78 mg, 0.054 mmol, 1.00 equiv) in N, N-dimethylformamide (1 ml) with stirring at 0 ℃. The mixture was allowed to react at room temperature for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After completion of the reaction, 20 ml of water was added to quench the reaction, and the resulting mixture was extracted with ethyl acetate (20 ml × 3). Combining the organic phases, drying the organic phases with anhydrous sodium sulfate, and filtering to remove the drying agent; concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by preparative thin layer chromatography eluting with a 0% → 8% methanol/dichloromethane mobile phase gradient to give compound 47-4a (white solid, 46 mg, 62% yield). MS (ESI, m/z) 1246.6[ m + H ], [ M ], [ phi ], [ H ] ] ⁺ 。

Step 6

To a solution of compound 47-4a (46 mg, 0.035 mmol, 1.00 eq) in methanol (1 ml) was added dropwise a solution of hydrochloric acid in 1, 4-dioxane (4 mol/l, 1 ml) with stirring at 0 ℃. The mixture was allowed to react at room temperature for 1 hour, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. Crude product is passed throughPurifying by high performance liquid chromatography with Sunfire per C18 Column,30 × 150 mm, 5 μm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml/min; elution was performed with 5% → 5% B in 1.5 minutes, 5% → 16% B in 2 minutes, and 16% → 30% B in 7 minutes; detector UV 220 nm; compound 47 (white solid, 16.8 mg, 41% yield) was obtained. MS (ESI, m/z) 1102.5[ 2 ], [ M + H ]] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ8.98(s,1H),8.58–8.55(m,1H),8.24(s,1H),7.98–7.96(m,1H),7.87(d,J＝9.6Hz,1H),7.55–7.45(m,2H),7.42–7.36(m,5H),7.15(d,J＝2.4Hz,1H),4.53(d,J＝9.6Hz,1H),4.45–4.39(m,2H),4.35–4.30(m,2H),4.27–4.19(m,3H),4.04–3.99(m,1H),3.96(s,1H),3.67–3.63(m,1H),3.62–3.58(m,4H),3.56–3.52(m,2H),3.50–3.46(m,3H),3.39–3.36(m,2H),3.04–3.00(m,1H),2.91–2.84(m,1H),2.82–2.75(m,1H),2.56–2.52(m,1H),2.43(s,3H),2.37–2.23(m,2H),2.19–2.13(m,1H),2.05–2.00(m,1H),1.92–1.85(m,2H),1.79–1.58(m,9H),0.89(s,9H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-110.19,-118.96,-124.73。

Example 48

(2S, 4R) -1- ((S) -2- (3- (2- ((2R, 6R, 7aS) -7a- ((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl) -6, 8-difluoroquinazolin-2-yl) oxy) methyl) -6-fluorohexahydro-1H-pyrrolizin-2-yl) ethoxy) propylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 48

Step 1

Compound 3-buten-1-ol (17.13 g, 225.62 mmol, 1.1 equiv) was dissolved in 300 ml of tetrahydrofuran under stirring at 0 ℃ under nitrogen protection, sodium hydride (60%, 24.61 g, 615.33 mmol, 3.0 equiv) was added thereto in portions, and after the mixture was reacted at 0 ℃ for 30 minutes, bromoacetic acid (30 g, 205.11 mmol, 1.0 equiv) was slowly added dropwise to the above mixed system. The mixture was allowed to react at 70 ℃ for 2 hours, and the progress of the reaction was monitored by thin layer chromatography. After the reaction is finished, the reaction liquid is cooled to room temperature, poured into an aqueous solution for quenching, the pH value is adjusted to 1-3 by using 2 mol of hydrochloric acid, ethyl acetate is used for extraction, organic phases are combined and dried, drying agents are removed by filtration, filtrate is concentrated to obtain a crude product, namely a compound 48-1 (yellow oil, 30 g, the yield is 56%), and the crude product is directly used for the next synthesis.

Step 2

Compound 48-1 (30 g, 218.99 mmol, 1.0 eq) was dissolved in 300 ml of anhydrous tetrahydrofuran under stirring at 0 deg.C under nitrogen, a solution of lithium aluminum hydride in tetrahydrofuran (2.5 mol per liter, 87.60 ml, 218.99 mmol, 1.0 eq) was slowly added dropwise thereto, and the resulting mixture was reacted at 25 deg.C for 2 hours, the progress of the reaction being monitored by thin layer chromatography. After the reaction is finished, cooling the reaction liquid to 0 ℃, slowly dripping water (20 ml), 20% sodium hydroxide (20 ml) and water (60 ml) into the reaction liquid in sequence, continuously stirring for 30 minutes after the water is added, filtering the mixture, and concentrating the filtrate to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a 0% → 50% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 48-2 (colorless oil, 5 g, yield 30%). ¹ H NMR(400MHz,CDCl ₃ )δ5.88-5.77(m,1H),5.20–4.99(m,2H),3.74-3.71(m,2H),3.63–3.48(m,4H),2.38-2.32(m,2H)。

Step 3

Compound 48-2 (5 g, 43.04 mmol, 1.0 eq) and carbon tetrabromide (15.66 g, 47.34 mmol, 1.1 eq) were dissolved in 50 ml of dichloromethane with stirring at 0 ℃ under nitrogen protection, triphenylphosphine (12.40 g, 47.34 mmol, 1.1 eq) was added slowly and the mixture was reacted at 0 ℃ for 1 hour with the progress of the reaction monitored by thin layer chromatography. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a 0% → 10% ethyl acetate/petroleum ether mobile phase gradient and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 48-3 (colorless oil, 3 g, 50% yield). ¹ H NMR(400MHz,CDCl ₃ )δ5.90–5.75(m,1H),5.14–5.00(m,2H),3.80–3.69(m,2H),3.60–3.51(m,2H),3.50–3.42(m,2H),2.41–2.31(m,2H)。

Step 4

Compound 48-4 (prepared in reference to WO2021041671, 2 g, 8.82 mmol, 1.0 eq), anhydrous methanol (0.57 g, 17.65 mmol, 2 eq) and 20 ml of anhydrous tetrahydrofuran were added sequentially to the reaction flask at 0 ℃ under nitrogen protection with stirring, followed by addition of lithium borohydride (0.40 g, 17.65 mmol, 2 eq) in portions to the above solution, and the resulting mixture was reacted at 25 ℃ for 2 hours, the progress of the reaction being monitored by thin layer chromatography. After the reaction was completed, the reaction solution was quenched by pouring into a saturated sodium bicarbonate solution, extracted with a mixed solvent of chloroform and isopropanol (3/1), and the organic phase was concentrated to obtain a crude product, compound 48-5 (colorless oil, 1.5 g, yield 90%). MS (ESI, m/z) 174.1[ 2 ], [ M ] +H ] ⁺ . The crude product was used directly in the next synthesis.

Step 5

Under the condition of stirring at 0 ℃ under the protection of nitrogen, the compound 48-5 (1) is added into the reaction bottle in sequence.5 g, 8.22 mmol, 1.0 eq), imidazole (0.88 g, 12.34 mmol, 1.5 eq) and 20 ml of dichloromethane, to which tert-butyldiphenylchlorosilane (3.09 g, 10.69 mmol, 1.3 eq) is then slowly added, and the resulting mixture is reacted at 25 ℃ for 2 hours, the course of the reaction being monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was quenched by pouring into saturated sodium bicarbonate solution, extracted with dichloromethane (20 ml x 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 5% methanol/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 48-5 (white solid, 3 g, 84% yield). MS (ESI, m/z) 412.2[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(400MHz,CDCl ₃ )δ7.64–7.61(m,4H),7.46–7.37(m,6H),5.33-5.18(m,1H),4.18–4.09(m,1H),3.57(d,J＝10.0Hz,1H),3.43(d,J＝10.0Hz,1H),3.15–3.02(m,1H),2.76–2.67(m,1H),2.42–2.12(m,3H),2.04–1.92(m,2H),1.05(s,9H)。

Step 6

After the mixture was reacted at-78 ℃ for 30 minutes under nitrogen protection with stirring, compound 48-6 (2.6 g, 6.32 mmol, 1.0 eq) was dissolved in 30 ml of tetrahydrofuran and a solution of lithium diisopropylamide in tetrahydrofuran (1 mol per liter, 8.2 ml, 8.20 mmol, 1.3 eq) was slowly added dropwise, a solution of 48-3 (1.46 g, 8.20 mmol, 1.3 eq) in anhydrous tetrahydrofuran (2 ml) was slowly added dropwise to the above system and the reaction was slowly warmed to room temperature after completion of the addition, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction is finished, pouring the reaction liquid into saturated ammonium chloride solution for quenching, extracting with ethyl acetate for three times, combining organic phases, drying the organic phases with anhydrous sodium sulfate, filtering to remove a drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel using 0% → 30% ethyl acetate Petroleum ether mobile phase gradient elution, the resulting fraction was subjected to reduced pressure rotary evaporation to remove the solvent, yielding compound 48-7 (white solid, 1.9 g, 55% yield). MS (ESI, m/z) 510.2[ 2 ], [ M ] +H] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.65–7.61(m,4H),7.45–7.39(m,6H),5.87–5.69(m,1H),5.36–5.18(m,1H),5.10–4.93(m,2H),4.25–4.07(m,1H),3.52–3.39(m,5H),3.19–3.01(m,1H),2.97–2.82(m,1H),2.41–2.14(m,5H),2.03–1.87(m,1H),1.77–1.68(m,1H),1.59–1.44(m,2H),1.04(s,9H)。

Step 7

The compound 48-7 (1.9 g) obtained in step 6 of this example was subjected to isomer separation by supercritical liquid chromatography: chiral column CHIRALPAK IA,3 × 25 cm, 5 microns; a mobile phase A: supercritical carbon dioxide, mobile phase B: methanol; flow rate: 50 ml/min; column temperature: 35 ℃; eluting with 15% mobile phase B; detector UV225 nm, resulting in two products. The product of shorter residence time (6.83 minutes) was compound 48-7a (colorless oil, 1.4 g, 80% recovery), compound 48-7a] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.69–7.57(m,4H),7.51–7.32(m,6H),5.86–5.69(m,1H),5.38–5.13(m,1H),5.09–4.89(m,2H),4.24–4.05(m,1H),3.59(d,J＝10.3Hz,1H),3.54–3.38(m,5H),3.22–3.00(m,1H),2.97–2.81(m,1H),2.45–2.11(m,5H),2.05–1.91(m,1H),1.76–1.66(m,1H),1.57–1.43(m,1H),1.04(s,9H)； ¹⁹ F NMR(282MHz,CDCl ₃ ) Delta-174.00. The product with longer retention time (10.02 min) was compound 48-7b (colorless oil, 270 mg, recovery 16%), compound 48-7b] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.67–7.56(m,4H),7.48–7.35(m,6H),5.86–5.67(m,1H),5.47–5.21(m,1H),5.10–4.93(m,2H),4.21–4.01(m,1H),3.59–3.53(m,1H),3.52–3.36(m,5H),3.36–3.20(m,1H),2.75–2.63(m,1H),2.58–2.40(m,1H),2.32–2.12(m,3H),2.08–1.89(m,2H),1.81–1.74(m,1H),1.65–1.51(m,1H),1.04(s,9H)； ¹⁹ F NMR(282MHz,CDCl ₃ )δ-170.22。

Step 8

Compound 48-7a (600 mg, 1.11 mmol, 1.0 eq) was dissolved in 6 ml of tetrahydrofuran at 0 ℃ with stirring under nitrogen protection, and a solution of lithium aluminum hydride in tetrahydrofuran (1 mol per liter, 1.67 ml, 1.67 mmol, 1.5 eq) was added slowly dropwise thereto, and the mixture was reacted at 60 ℃ for 2 hours, the course of the reaction being monitored by liquid chromatography and thin layer chromatography. After the reaction, the reaction solution was cooled to 0 ℃, water (1 ml), a 20% aqueous solution of sodium hydroxide (1 ml) and water (3 ml) were slowly added thereto in this order to quench, the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 10% methanolic ammonia/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 48-8a (colorless oil, 210 mg, 69% yield). MS (ESI, m/z) 258.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ5.83–5.66(m,1H),5.27–4.91(m,3H),3.42–3.17(m,7H),3.13–3.03(m,1H),2.99–2.79(m,1H),2.60–2.53(m,1H),2.31–2.08(m,3H),2.06–1.79(m,3H),1.66–1.50(m,3H)。

Step 9

Compound 46-1 (600 mg, 0.739 mmol, 1 eq), compound 48-8a (200 mg, 0.739 mmol, 1 eq), cesium carbonate (506.64 mg, 1.478 mmol, 2 eq), triethylenediamine (17.44 mg, 0.148 mmol, 0.2 eq), and anhydrous N, N-dimethylformamide (5 ml) were added to the flask, in that order, with stirring, at 25 degrees celsius under nitrogen protection. The resulting mixture was reacted at 80 ℃ for 2.5 hours, and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. Reaction(s) ofAfter completion, the reaction mixture was diluted with 50 ml of water, the aqueous layer was extracted with ethyl acetate (50 ml × 3), and after combining, the organic layer was washed with saturated brine (50 ml × 3) and dried over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane and the resulting fraction was concentrated under reduced pressure to remove the solvent and afford compound 48-9a (white solid, 580 mg, 63% yield). MS (ESI, m/z) 864.4[ m + H ]] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ7.71–7.65(m,1H),7.52(d,J＝3.0Hz,1H),7.40–7.26(m,1H),7.26–7.22(m,1H),7.11(d,J＝3.0Hz,1H),5.84–5.56(m,3H),5.29(s,2H),5.10–4.94(m,2H),4.50-4.0(m,6H),3.70–3.50(m,5H),3.48–3.34(m,5H),2.77–2.53(m,2H),2.46–2.24(m,6H),2.14–1.83(m,7H),1.52(s,9H),1.32–1.22(m,2H),0.86–0.80(m,3H)。

Step 10

The compound 48-9a (580 mg) obtained in step 9 of this example was subjected to chiral resolution by supercritical liquid chromatography: CHIRAL column CHIRAL ART amide-SA, 3 × 25 cm, 5 microns; mobile phase A: supercritical carbon dioxide, mobile phase B: isopropanol (0.5%, 2 mol/l ammonia methanol); flow rate: 100 ml/min; column temperature: 35 ℃; eluting with 40% mobile phase B; detector UV222 nm, resulting in two products. The product at shorter retention time (2.75 min) was compound 48-9aa (white solid, 220 mg, 38% recovery); MS (ESI, m/z) 864.4[ m + H ] ] ⁺ (ii) a The product of longer retention time (4.37 min) was compound 48-9ab (white solid, 230 mg, 39% yield); MS (ESI, m/z) 864.4[ m + H ]] ⁺ 。

Step 11

Stirring at 0 deg.C, adding into a reaction flaskCompound 48-9aa (120 mg, 0.132 mmol, 1 eq), sodium periodate (178.24 mg, 0.792 mmol, 6 eq), carbon tetrachloride (0.6 ml), acetonitrile (0.6 ml) were added successively, and then a solution of ruthenium trichloride (3.13 mg, 0.013 mmol, 0.1 eq) in water (0.9 ml) was slowly added dropwise thereto. The mixture was reacted at 0 ℃ for 1 hour, and the reaction was monitored by liquid mass. After the reaction is finished, directly concentrating the reaction solution to obtain a crude product. The crude product obtained was purified by reverse phase chromatography (C18 column), mobile phase a: water (0.1% ammonium bicarbonate); mobile phase B, methanol, eluting with 5% → 95% of phase B in 25 minutes; detector UV254/220 nm; compound 48-10aa (white solid, 70 mg, 54% yield) was obtained. MS (ESI, m/z) 882.4[ m + H ]] ⁺ 。

Step 12

To a solution of compound 48-11aa (70 mg, 0.071 mmol, 1.00 eq) in N, N-dimethylformamide (1.5 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethyluronium hexafluorophosphate (37.17 mg, 0.092 mmol, 1.3 eq) with stirring at room temperature. After the mixture was stirred at 25 ℃ for 10 minutes, the reaction mixture was added with (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl group ]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (40.11 mg, 0.085 mmol, 1.2 eq) and N, N-diisopropylethylamine (38.87 mg, 0.284 mmol, 4 eq). The mixture was stirred at 25 ℃ for 1 hour and the progress of the reaction was monitored by liquid mass. After completion of the reaction, the crude product obtained was purified by reverse phase chromatography (C18 column), mobile phase a: water (0.1% ammonium bicarbonate); mobile phase B, methanol, eluting with 10% → 95% of phase B in 25 minutes; detector UV254/220 nm; compound 48-11aa (white solid, 80 mg, 81% yield) was obtained. MS (ESI, m/z) 1308.6[ 2 ], [ M ] +H] ⁺ 。

Step 13

To a solution of compound 48-11aa (80 mg, 0.058 mmol, 1.00 eq) in methanol (2 ml) was added dropwise a solution of 1, 4-dioxane (4 mol/l, 2 ml) of hydrochloric acid with stirring at 0 ℃. The mixture was allowed to react at room temperature for 2 hours, and the progress of the reaction was monitored by liquid phase. After the reaction is finished, the reaction solution is concentrated to obtain a crude product. The crude product obtained was purified by reverse phase chromatography (C18 column), mobile phase a: water (0.1% ammonium bicarbonate); mobile phase B, methanol, eluted with 70% → 95% of phase B in 30 minutes; detector UV254/220 nm. Compound 48 was obtained (white solid, 50 mg, 70% yield). MS (ESI, m/z) 1164.5[ m + H ] ] ⁺ 。 ¹ H NMR(400MHz,DMSO-d ₆ )δ10.02(s,1H),8.98(s,1H),8.36(d,J＝7.8Hz,1H),7.89–7.74(m,2H),7.70–7.62(m,1H),7.45–7.30(m,6H),7.01(d,J＝2.6Hz,1H),5.36–5.22(m,1H),5.11(s,1H),4.97–4.84(m,1H),4.51(d,J＝9.3Hz,1H),4.47–4.37(m,1H),4.33–4.19(m,3H),4.12–4.01(m,1H),4.00–3.91(m,1H),3.64–3.43(m,8H),3.42–3.34(m,2H),3.30–3.25(m,1H),3.24–3.17(m,1H),3.12–2.87(m,2H),2.62–2.54(m,1H),2.45(s,3H),2.39–1.94(m,8H),1.82–1.73(m,1H),1.65(s,4H),1.58–1.50(m,2H),1.50–1.41(m,1H),1.37(d,J＝7.0Hz,3H),0.90(s,9H),0.79–0.70(m,3H)； ¹⁹ F NMR(377MHz,DMSO-d ₆ )δ-118.64,-119.23,-123.86,-171.35。

Example 49

(2S, 4R) -1- ((S) -2- (3- (2- ((2S, 6R, 7aS) -7a- ((S or R) -4- ((1R, 5S) -3, 8-diazacyclo [3.2.1] octan-3-yl) -7- (8-ethyl-7-fluoro-3-hydroxynaphthalen-1-yl) -6, 8-difluoroquinazolin-2-yl) oxy) methyl) -6-fluorohexahydro-1H-pyrrolizin-2-yl) ethoxy) propylamino) -3, 3-dimethylbutyryl) -4-hydroxy-N- ((S) -1- (4- (4-methylthiazol-5-yl) phenyl) ethyl) pyrrolidine-2-carboxamide 49

Step 1

Compound 48-7b (270 mg, 0.503 mmol, 1.0 eq) was dissolved in 3 ml of tetrahydrofuran at 0 ℃ with stirring under nitrogen, and then a solution of lithium aluminum hydride in tetrahydrofuran (1 mol per liter, 0.75 ml, 0.75 mmol, 1.5 eq) was slowly added dropwise thereto, and the resulting mixture was reacted at 60 ℃ for 16 hours, the course of the reaction being monitored by liquid mass and thin layer chromatography. After the reaction, the reaction solution was cooled to 0 ℃, water (1 ml), a 20% aqueous solution of sodium hydroxide (1 ml) and water (3 ml) were slowly added thereto in this order to quench, the mixture was filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by column chromatography on silica gel eluting with a mobile phase gradient of 0% → 10% methanolic ammonia/dichloromethane and the resulting fraction was freed from the solvent by rotary evaporation under reduced pressure to give compound 49-1 (colorless oil, 60 mg, 44% yield). MS (ESI, m/z) 258.2[ 2 ], [ M ] +H ] ⁺ ； ¹ H NMR(300MHz,CDCl ₃ )δ5.97–5.69(m,1H),5.39–4.95(m,3H),3.53–3.30(m,5H),3.29–3.19(m,1H),3.15–2.99(m,2H),2.81–2.69(m,1H),2.63–2.45(m,1H),2.41–2.20(m,3H),2.18–1.89(m,3H),1.70–1.56(m,2H),1.55–1.43(m,1H)。

Step 2

Compound 46-1 (120 mg, 0.177 mmol, 1 eq), compound 49-1 (48.02 mg, 0.177 mmol, 1 eq), cesium carbonate (121.59 mg, 0.354 mmol, 2 eq), triethylenediamine (4.19 mg, 0.035 mmol, 0.2 eq), and anhydrous N, N-dimethylformamide (2 ml) were added to the flask sequentially with stirring at 25 degrees celsius under nitrogen. The resulting mixture was stirred at 80 ℃ for 3 hours and the progress of the reaction was monitored by liquid chromatography and thin layer chromatography. After the reaction was complete, 20 ml of water was addedThe reaction solution was diluted, and the resulting mixture was extracted with ethyl acetate (20 ml × 3), and after combining, the organic layer was washed with saturated brine (20 ml × 3) and dried over anhydrous sodium sulfate. Filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography eluting with a mobile phase gradient of 0% → 10% methanol/dichloromethane, and the resulting fraction was concentrated under reduced pressure to remove the solvent, yielding compound 49-2 (white solid, 120 mg, 73% yield). MS (ESI, m/z) 864.4[ 2 ], [ M ] +H] ⁺ 。

Step 3

Chiral resolution of compound 49-2 (120 mg) obtained in step 2 of this example was performed by supercritical liquid chromatography: CHIRAL column CHIRAL ART amide-SA, 3 × 25 cm, 5 microns; mobile phase A: n-hexane (10 mmol/l ammonia methanol), mobile phase B: ethanol; flow rate: 20 ml/min; elution with 30% mobile phase B over 11 minutes; detector UV230/220 nm, resulting in two products. The product with shorter retention time (5.39 min) is compound 49-2a (white solid, 58 mg, recovery 48%), MS (ESI, m/z): 864.4[ m + H ] ] ⁺ (ii) a The product with longer retention time (7.33 min) is compound 49-2b (white solid, 50 mg, 41% recovery), MS (ESI, m/z) 864.4[ m + h ]] ⁺ 。

Step 4

Compound 49-2a (60 mg, 0.064 mmol, 1 eq), sodium periodate (86.15 mg, 0.384 mmol, 6 eq), carbon tetrachloride (0.5 ml), acetonitrile (0.5 ml) were added to the flask in succession with stirring at 0 ℃, to which was slowly added dropwise a solution of ruthenium trichloride (1.51 mg, 0.006 mmol, 0.1 eq) in water (0.75 ml). The mixture was reacted at 0 ℃ for 1 hour, and the progress of the reaction was monitored by liquid mass. After the reaction is finished, the reaction liquid is poured into waterQuenching, extracting with chloroform and isopropanol mixed solvent (3/1) for three times, combining, drying the organic phase with anhydrous sodium sulfate, filtering to remove the drying agent, and concentrating the filtrate under reduced pressure to obtain a crude product. The crude product obtained was purified by reverse phase chromatography (C18 column), mobile phase a: water (0.1% ammonium bicarbonate); mobile phase B, methanol, eluting with 5% → 95% of phase B in 25 minutes; detector UV254/220 nm; compound 49-3a was obtained (white solid, 20 mg, 34% yield). MS (ESI, m/z) 882.4[ m + H ]] ⁺ 。

Step 5

To a solution of compound 49-3a (70 mg, 0.022 mmol, 1.00 eq) in N, N-dimethylformamide (1 ml) was added 2- (7-azobenzotriazol) -N, N' -tetramethylurea hexafluorophosphate (10.35 mg, 0.026 mmol, 1.3 eq) with stirring at room temperature. After the mixture was stirred at 25 ℃ for 10 minutes to react, to the reaction solution were added (2S, 4R) -1- [ (2S) -2-amino-3, 3-dimethylbutyryl) in this order ]-4-hydroxy-N- [ (1S) -1- [4- (4-methyl-1, 3-thiazol-5-yl) phenyl ] methyl]Ethyl radical]Pyrrolidine-2-carboxamide hydrochloride (11.09 mg, 0.024 mmol, 1.1 eq) and N, N-diisopropylethylamine (11.72 mg, 0.088 mmol, 4 eq). The resulting mixture was stirred at 25 ℃ for 1 hour, and the progress of the reaction was monitored by liquid phase. After completion of the reaction, the crude product obtained was purified by reverse phase chromatography (C18 column), mobile phase a: water (0.1% ammonium bicarbonate); mobile phase B, methanol, eluting with 10% → 95% of phase B in 25 minutes; detector UV254/220 nm; the resulting fraction was concentrated under reduced pressure to give compound 49-4a (white solid, 20 mg, 84% yield). MS (ESI, m/z) 1308.6[ 2 ], [ M ] +H] ⁺ 。

Step 6

Adding to compound 49 under stirring at 0 deg.C4a (20 mg, 0.015 mmol, 1.00 eq) in methanol (1 ml) was added dropwise a solution of 1, 4-dioxane (4 mol/l, 1 ml) in hydrochloric acid. The mixture was allowed to react at room temperature for 2 hours, and the progress of the reaction was monitored by liquid mass. After the reaction is finished, concentrating the reaction liquid to obtain a crude product. The crude product obtained was purified by reverse phase chromatography (C18 column), mobile phase a: water (0.1% ammonium bicarbonate); mobile phase B, methanol, eluted with 70% → 95% of phase B in 30 minutes; detector UV254/220 nm; the resulting fraction was concentrated under reduced pressure to give compound 49 (white solid, 5.5 mg, 30% yield). MS (ESI, m/z) 1164.5[ m + H ] ] ⁺ 。 ¹ H NMR(400MHz,CD ₃ OD)δ8.86(s,1H),7.69–7.55(m,2H),7.47–7.33(m,4H),7.31–7.17(m,2H),6.96(d,J＝2.7Hz,1H),5.33–5.20(m,1H),5.04–4.96(m,1H),4.64(s,1H),4.60–4.53(m,1H),4.50–4.38(m,3H),4.30–4.20(m,2H),3.85(d,J＝11.0Hz,1H),3.77–3.70(m,1H),3.70–3.64(m,2H),3.64–3.54(m,4H),3.53–3.43(m,3H),3.39–3.34(m,1H),3.17–3.04(m,2H),2.92–2.78(m,1H),2.68–2.49(m,4H),2.48–2.41(m,4H),2.26–2.17(m,1H),2.17–2.03(m,2H),1.97–1.89(m,1H),1.88–1.81(m,3H),1.75–1.63(m,3H),1.59–1.44(m,3H),1.36–1.30(m,1H),1.03(s,9H),0.84–0.69(m,3H)； ¹⁹ F NMR(377MHz,CD ₃ OD)δ-118.75,-121.17,-124.29,-174.68。

Effect example 1

1. Purpose of the experiment

The inhibitory capacity of the small-molecule compound on the binding activity of KRAS-G12D and SOS1 is detected by a drug screening system based on the binding of KRAS _ G12D and SOS 1.

2. Experimental material and instrument and equipment

3. Experimental methods

3.1 Experimental procedure:

a) BI-2852 as a positive control, stock solution was diluted first, 3-fold, diluted 10+0 spots. The first point of dilution of the test compound is also its stock solution, 3-fold diluted, diluted 11+0 points. Transfer 0.2 μ L of the compound solution diluted in a gradient to 384-well plates with 2 replicates of each compound in an Echo, with a final DMSO concentration of 1%. Centrifuge at 1000rpm/min for 1min. The final Reference concentration is 100, 33.33, 11.11,3.70,1.23,0.412,0.137,0.046,0.015,0.005,0 μ M. The final concentrations of test compounds are 200, 66.67, 22.22,7.41,2.47,0.27,0.091,0.03,0.0152,0.01, 0. Mu.M.

b) KRAS _ G12D in the kit and GTP with the final concentration of 10 μ M are prepared together in a diluent, 5 μ L is transferred to a 384 reaction plate, 1000rpm/min and centrifuged for 1min,

c) Transfer 5. Mu.L of SOS1 mixture to 384 reaction plates, 1000rpm/min, centrifuge for 1min, incubate for 15min at 25 ℃.

d) Transfer 10. Mu.L of assay mix to 384 reaction plates, 1000rpm/min, centrifuge for 1min, and incubate overnight at 4 ℃.

e) The excitation wavelength of 665nm and the emission wavelength of 615nm were read using an Envision multifunction plate reader. 665/615Ratio signal intensity is used to characterize the degree of activity of the enzyme.

f) The raw data was analyzed.

3.2 Experimental data processing method:

compound IC50 was fitted by Graphpad Prism 8 nonlinear regression equation:

negative control: DMSO (dimethylsulfoxide)

Positive control: 100 μ M BI-2852

The IC of the compound was obtained using the following non-linear fit equation ₅₀ (median inhibitory concentration):

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*HillSlope))

x log value of Compound concentration

Y:665/615Ratio

Effect example B

1. Purpose of the experiment

The inhibition ability of the small molecule compound on the binding activity of KRAS _ G12D and cRAF is detected by a drug screening system based on the binding of KRAS _ G12D and cRAF.

2. Experimental material and instrument and equipment

Reagent	Brand	Goods number
			KRAS-G12D/cRAF binding kits	Cisbio	63ADK000CB21PEG
GTP	Sigma	V900868
			Consumable material	Brand	Goods number
Topseal A	PerkinElmer	E5341
			384-Well Polypropylene microplate	labcyte	PP-0200
96Well Plates	Nunc	249944
			384-well plates	Corning	CLS4514
Instrument for measuring the position of a moving object	Brand	Goods number
			Envision	Perkin Elmer	2104
Centrifuge	Eppendorf	5810R
			Multi-channel pipettes	Eppendorf/Sartorius	/
Echo	Labcyte	/

3. Experimental methods

3.1 Experimental procedure:

a) BI-2852 as a positive control, stock solution was diluted first, 3-fold, diluted 10+0 spots. The first spot of the same dilution of test compound is its stock solution, 3-fold diluted, diluted 11+0 spots. Transfer 0.2 μ L of the compound solution diluted in a gradient to 384-well plates with 2 replicates of each compound in an Echo, with a final DMSO concentration of 1%. Centrifuge at 1000rpm/min for 1min. The final concentration of the positive control was 100, 33.33, 11.11,3.70,1.23,0.412,0.137,0.046,0.015,0.005, 0. Mu.M. The final concentrations of test compounds are 200, 66.67, 22.22,7.41,2.47,0.27,0.091,0.03,0.0152,0.01, 0. Mu.M.

c) Transfer 5. Mu.L of the cRAF mixture to 384 reaction plates at 1000rpm/min, centrifuge for 1min and incubate for 15min at 25 ℃.

f) The raw data was analyzed.

3.2 Experimental data processing method:

fitting Compound IC by Graphpad Prism 8 nonlinear regression equation ₅₀ ：

Negative control: DMSO (dimethylsulfoxide)

Positive control: 100 μ M BI-2852

The IC50 (half maximal inhibitory concentration) of the compound was obtained using the following non-linear fit equation:

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope))

x log value of Compound concentration

Y:665/615Ratio

Effect example C

Purpose of the experiment

The inhibitory ability of small molecule compounds on the binding activity of KRAS _ WT to SOS1 was tested by a drug screening system based on KRAS _ WT binding to SOS 1.

2 Experimental materials and instruments and equipment

Reagent	Brand	Goods number
			KRAS-WT/SOS1 binding kits	Cisbio	63ADK000CB15PEH
GTP	Sigma	V900868
			Consumable material	Brand	Goods number
Topseal A	PerkinElmer	E5341
			384-Well Polypropylene microplate	labcyte	PP-0200
96Well Plates	Nunc	249944
			384-well plates	Corning	CLS4514
Instrument for measuring the position of a moving object	Brand	Goods number
			Envision	Perkin Elmer	2104
Centrifuge	Eppendorf	5810R
			Multi-channel pipettes	Eppendorf/Sartorius	/
Echo	Labcyte	/

3 Experimental methods

3.1 Experimental procedure:

a) BI-2852 as a positive control, stock solution was the first point of dilution, 3-fold dilution, dilution 10+0 point. The first point of dilution of the test compound is also its stock solution, 3-fold diluted, diluted 11+0 points. Transfer 0.2 μ L of the compound solution diluted in a gradient to 384-well plates with 2 replicates of each compound in an Echo, with a final DMSO concentration of 1%. Centrifuge at 1000rpm/min for 1min. The final concentration of the positive control is 100, 33.33, 11.11,3.70,1.23,0.412,0.137,0.046,0.015,0.005,0 μ M. The final concentrations of test compounds are 200, 66.67, 22.22,7.41,2.47,0.27,0.091,0.03,0.0152,0.01, 0. Mu.M.

b) KRAS _ WT and GTP with the final concentration of 10 μ M in the kit are prepared together in diluent, transferred into a 384 reaction plate with 5 μ L, centrifuged at 1000rpm/min for 1min,

e) The excitation wavelength 665nm and the emission wavelength 615nm were read using an Envision multifunction plate reader. 665/615Ratio signal intensity is used to characterize the degree of activity of the enzyme.

f) The raw data was analyzed.

3.2 Experimental data processing method:

Negative control: DMSO (dimethylsulfoxide)

Positive control: 100 μ M BI-2852

IC of the compound was obtained using the following non-linear fit equation ₅₀ (median inhibitory concentration):

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*HillSlope))

x log value of Compound concentration

Y:665/615Ratio

The experimental results are as follows: the results of the tests of examples A-C are shown in Table 1:

the structural formula of BI-2852 is

BI-2852 was prepared from Supplementary Information of the reference (doi: 10.1073/pnas.1904529116).

Effect example 2KRAS-G12D PROTAC molecular Degradation test (Degradation Assay)

2.1 purpose of the experiment

The KRAS protein degradation targeting ability of KRAS-G12D molecules was tested by Jess WB assay on the KRAS protein degradation in PANC-1 and GP2D cells (KRAS-G12D mutants).

2.2 Experimental materials and instruments and Equipment

2.2 Experimental methods

1. After adherent culture of the recovered PANC-1 and GP2D cells for 1-2 passages, the cells were inoculated into 12-well plates, and the plates were placed at 37 ℃ for 5% CO ₂ Incubate overnight. The compound of example 25 is added at a concentration of 30uM,1/3 dilution, 7+0dose,48h.

2. The medium was aspirated off, cells were washed once with 1mL 1X PBS, 300. Mu.L of Tryple (Gibco) was added to each well, digestion was stopped at 37 ℃ for 5min, 900. Mu.L of medium was added to each well, cells were transferred to a 1.5mL centrifuge tube and centrifuged at 2000rpm at 4 ℃ for 5min to collect cells. Cells were washed once with 1mL 1X PBS and harvested by centrifugation at 2000rpm for 5min at 4 ℃.

3. Adding a proper amount of lysis solution containing 1x PMSF according to the cell amount, uniformly mixing, placing on ice for lysis for 30min, centrifuging at 13000rpm at 4 ℃ for 20min, and transferring the centrifuged supernatant to a new centrifuge tube.

4. Taking 20 μ L of Bovine Serum Albumin (BSA) with different known concentrations to prepare a protein quantitative standard curve; diluting the sample protein to be detected by 10 times, and adding 20 mu L of the diluted sample protein into a corresponding detection hole; get Pierce ^TM BCA Protein Assay Kit BCA quantitative reagent, 200 mu L/well, avoids generating air bubbles, and detects the Optical Density (OD) value under the condition of 562nm wavelength after incubation for 30min at 37 ℃ in the dark.

5. Adding a proper amount of 0.1 Xloading buffer (sample buffer) into a protein sample, heating in a boiling water bath for 5min to fully denature the protein, and quantitatively analyzing the degradation rate of the KRAS-G12D protein by a Jess WB system, wherein the primary Antibody is KRAS mouse Antibody (Lsbio) or GAPDH mouse mAb (CST).

2.3 Experimental results:

the compounds of the invention were tested for their degradation activity in cells based on the same WB degradation assay, with the results being determined by DC ₅₀ And Dmax, where Dmax is the maximum level of protein degradation observed, DC ₅₀ Is the compound concentration required to achieve 50% of Dmax; the results are shown in Table 2:

TABLE 2 degradation Activity data for Compounds

Example 25 molecules showed about 50% degradation of KRAS G12D protein on PANC-1 cells and about 40% degradation of KRAS G12D protein on GP2D cells at a concentration of 30 μ M.

Example 45 molecules DC50 to KRAS protein in PANC-1 cells was 53.02nM, dmax was 89%; DC50 to KRAS protein in GP2D cells was 807nM, dmax was 83%.

Claims

1. A compound of formula Ia, ib, or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof, or an isotopic compound thereof:

wherein R is ^1a 、R ^1b And R ^1c Independently is C ₆ -C ₁₈ Aryl radical, by R ^1a-1 Substituted C ₆ -C ₁₈ Aryl or 5-10 heteroaryl, wherein the heteroatom in the 5-10 heteroaryl is one or more of N, O or S independently, and the number of the heteroatoms is 1-4;

R ^1a-1 independently of one another is hydroxy, halogen, C ₁ -C ₁₂ Alkyl or C ₂ -C ₆ An alkynyl group;

R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c and R ^5c Independently isH or halogen;

R ^3a is 7-12 membered bridged heterocycloalkyl, the heteroatoms in the bridged heterocycloalkyl are independently one or more of N, O or S, and the number of the heteroatoms is 1-4;

R ^4c is H, 3-6 membered heterocycloalkyl or is-N (R) ^4c-1 R ^4c-2 ) Substituted 3-6 membered heterocycloalkyl, the heteroatom of said 3-6 membered heterocycloalkyl and the substituted-N (R) ^4c-1 R ^4c-2 ) The heteroatom in the substituted 3-6 membered heterocycloalkyl is independently one or more of N, O or S, and the number of the heteroatoms is 1-3;

R ^4c-1 and R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

A ^a and A ^b Independently of one another is CR ^5a Or N; r is ^5a Independently of each other H, C ₁ -C ₄ Alkyl or halogen;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl, -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) — or absent; said 3-9 membered heterocycloalkyl group and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 The heteroatom in the' is one or more of N, O or S independently, and the number of the heteroatom is 1-3;

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ Alkyl or halogen;

Y ¹ is O, -NR ⁸ Or is absent;

n2 is 0, 1, 2 or 3;

Y ² is O, -NR ⁹ Or is absent;

R ⁸ and R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 0, 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

L ^a 、L ^b and L ^c Is a linking group;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand.

2. The compound of formula Ia, ib, or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof, or an isotopic compound thereof, of claim 1, wherein the compound of formula Ia, ib, or Ic, the pharmaceutically acceptable salt thereof, the solvate thereof, the stereoisomer thereof, the tautomer thereof, the prodrug thereof, the metabolite thereof, or the isotopic compound thereof, satisfies one or more of the following conditions:

(1)R ^1a 、R ^1b And R ^1c In (b), the C ₆ -C ₁₈ Aryl or said aryl is substituted by R ^1a-1 Substituted C ₆ -C ₁₈ C in aryl ₆ -C ₁₈ Aryl is phenyl, naphthyl, phenanthryl or anthracyl, preferably naphthyl;

(2)R ^1a 、R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c 、R ^5a and R ^5c Wherein said halogen is F, cl, br or I, e.g. F or Cl;

(3)R ^1a in (b), the C ₁ -C ₁₂ Alkyl is C ₁ -C ₆ Alkyl, which can also be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl;

(4)R ^1a in (b), the C ₂ -C ₆ Alkynyl is C ₂ -C ₄ Alkynyl, which can also be ethynyl;

(5)R ^3a wherein the 7-12 membered bridged heterocycloalkyl is a 7-or 8-membered bridged heterocycloalkyl group containing 2 heteroatoms N, and may be diazabicyclo [2.2.1 ]]Heptylalkyl, diazabicyclo [3.2.1]Octyl or diazabicyclo [2.2.2 ]]An octyl group;

(6)R ^3a wherein said 7-12 membered bridged heterocycloalkyl is attached to the pyrimidine ring through a heteroatom;

(7)R ^4c wherein said 3-to 6-membered heterocycloalkyl group is substituted with-N (R) ^4c-1 R ^4c-2 ) The 3-6 membered heterocycloalkyl group of the substituted 3-6 membered heterocycloalkyl group is azetidinyl, pyrrolidinyl or piperidinyl;

(8)R ^4c wherein said 3-to 6-membered heterocycloalkyl group is substituted with-N (R) ^4c-1 R ^4c-2 ) The 3-6 membered heterocycloalkyl of the substituted 3-6 membered heterocycloalkyl is attached to the pyrimidine ring through a heteroatom;

(9)R ^4c-1 and R ^4c-2 In (b), the C ₁ -C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl;

(10)M ¹ wherein the 3-9 membered heterocycloalkyl group is a 3-6 membered monocyclic heterocycloalkyl group, the heteroatoms are independently N and/or O, the number of the heteroatoms is 1 or 2, and the heteroatoms are further pyrrolidinyl;

(11)M ¹ in (2), the quilt M ^1-1 3-9 membered heterocycloalkyl of substituted 3-9 membered heterocycloalkyl is 3-6 membered monocyclic heterocycloalkyl or 8-9 membered fused ring hybrid alkyl, the heteroatoms are independently N and/or O, the number of heteroatoms is 1 or 2, further is pyrrolidinyl or hexahydro-1H-pyrrolizinyl;

(12)M ¹ in (b), the-3-to 9-membered heterocycloalkyl- (CH) ₂ ) _n1’ OC(＝O) The 3-to 9-membered heterocycloalkyl group in (E) is an 8-to 9-membered fused-ring hybrid alkyl group, the hetero atoms are independently N and/or O, the number of hetero atoms is 1 or 2, and further

(13)M ^1-1 In (b), the C ₁ -C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl;

(14)M ^1-1 wherein said halogen is F, cl, br or I, e.g., F;

(15) In the ring D, the 3-6 membered heterocyclic alkyl is azetidinyl, pyrrolidinyl or piperazine;

(16)R ^5a 、R ⁸ 、R ⁹ 、R ¹⁰ 、R ¹¹ and M ^1-1 In (b), the C ₁ -C ₄ Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, for example methyl or ethyl;

(17) In the ring E, the 3-6 membered heterocyclic alkyl is azetidinyl;

(18)L ^a 、L ^b and L ^c Independently is

e terminal and X ^a 、X ^b Or X ^c Is connected with terminal f and Q ^a 、Q ^b Or Q ^c Connecting;

n4, n6 and n7 are independently 0, 1 or 2;

n5 and n8 are independently any integer from 1 to 5, such as 1, 2, 3, 4 or 5;

Y ³ is NH, CH ₂ Or is absent;

n9 is any integer of 0-13;

n10 and n13 are independently 0, 1 or 2;

n11 and n12 are independently any integer from 1 to 7, for example 1, 2, 3, 4 or 5;

z is CH ₂ Or N;

Y ⁴ is-C (= O) or absent;

n14 and n15 are independently 0, 1, 2, 3, 4 or 5;

n16 and n19 are 0, 1 or 2;

n17, n18 and n20 are independently 0, 1, 2, 3, 4 or 5;

Z ¹ is CH or N;

(19)Q ^a 、Q ^b and Q ^c Independently is

3. The compound of formula Ia, ib, or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof, or an isotopic compound thereof of claim 2, wherein the compound of formula Ia, ib, or Ic, the pharmaceutically acceptable salt thereof, the solvate thereof, the stereoisomer thereof, the tautomer thereof, the prodrug thereof, the metabolite thereof, or the isotopic compound thereof, satisfies one or more of the following conditions:

(1) When R is ^1a 、R ^1b And R ^1c Independently is

When it is used, the

Is composed of

And/or

Wherein R is ⁶ And R ⁷ Independently of one another H, halogen, C ₁ -C ₁₂ Alkyl or C ₂ -C ₆ An alkynyl group;

(3)R ^3a wherein said 7-12 membered bridged heterocycloalkyl group is

(4)R ^4c In (b), the quilt is-N (R) ^4c-1 R ^4c-2 ) Substituted 3-6 membered heterocycloalkyl is substituted with 1-N (CH) ₃ ) ₂ ) Substituted azetidinyl;

(5)M ¹ in (1), the quilt M ^1-1 Substituted 3-9 membered heterocycloalkyl is pyrrolidinyl substituted by methyl or hexahydro-1H-pyrrolizinyl substituted by F, e.g.

The a terminal being linked to the alkylene group and the b terminal being linked to Y ¹ Connecting;

(6)Y ⁵ the 3-to 6-membered cycloalkyl group is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, for example cyclohexyl;

(7)Y ⁵ wherein the 3-to 6-membered heterocycloalkyl group is a 5-membered heterocycloalkyl group having 1 heteroatom of O or S, such as furyl;

(8)Y ⁵ in the 5-6 membered heteroaryl, the heteroatom is N, and the number is 3And 5-membered heteroaryl of (a), for example, 1-H,1,2, 3-triazolyl.

4. A compound of formula Ia, ib or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof or an isotopic compound thereof according to claim 3, wherein the compound of formula Ia, ib or Ic, the pharmaceutically acceptable salt thereof, the solvate thereof, the stereoisomer thereof, the tautomer thereof, the prodrug thereof, the metabolite thereof or the isotopic compound thereof satisfies one or more of the following conditions:

(1) Said

Is composed of

For example

(2)X ^a Is composed of

For example, in

(3)X ^b Is composed of

(4)L ^a 、L ^b And L ^c Independently is L ^a 、L ^b And L ^c Independently is

5. The compound of formula Ia, ib or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof or an isotopic compound thereof according to claim 1, wherein the compound of formula Ia, ib or Ic, the pharmaceutically acceptable salt thereof, the solvate thereof, the stereoisomer thereof, the tautomer thereof, the prodrug thereof, the metabolite thereof or the isotopic compound thereof satisfies one or more of the following conditions:

(1)R ⁷ is H or halogen;

(2)R ^2a 、R ^2b and R ^2c And R ^5c Independently is halogen;

(3) When A is ^a And A ^b Independently of one another is CR ^5a When R is ^5a Independently is halogen;

(4)X ^a is composed of

(5) n1 is 1 or 3;

(6) n2 is 0 or 1.

6. The compound of formula Ia, ib or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof or an isotopic compound thereof according to claim 1, wherein the compound of formula Ia, ib or Ic, the pharmaceutically acceptable salt thereof, the solvate thereof, the stereoisomer thereof, the tautomer thereof, the prodrug thereof, the metabolite thereof or the isotopic compound thereof satisfies any one of the following conditions:

(1)

Is composed of

M ¹ In the absence or 3-to 9-membered heterocycloalkyl, Y ¹ is-NR ⁸ ；

(2)

Is composed of

M ¹ Is as a quilt M ^1-1 Substituted 3-to 9-membered heterocycloalkyl, Y ¹ Is O;

(3)

is composed of

M ¹ Is 3-9 membered heterocycloalkyl, Y ¹ Is absent;

(4)

is composed of

(5)

Is composed of

(6)

Is composed of

(7)

Is composed of

7. A compound of formula Ia, ib or Ic, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof or an isotopic compound thereof according to claim 1, which is any one of schemes:

scheme 1:

the compound shown in formula Ia, ib or Ic, pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, metabolite or isotope compound thereof:

R ^1a-1 independently of one another, hydroxy, halogen, C ₁ -C ₁₂ Alkyl or C ₂ -C ₆ An alkynyl group;

R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c and R ^5c Independently H or halogen;

R ^3a Is 7-12 membered bridged heterocycloalkyl, wherein the heteroatom in the bridged heterocycloalkyl is independently one or more of N, O or S, and the number of the heteroatoms is 1-4;

R ^4c-1 and R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl, -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) — or absent; said 3-9 membered heterocycloalkyl and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 The heteroatom in the' is one or more of N, O or S independently, and the number of the heteroatom is 1 to 3;

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ An alkyl group;

Y ¹ is O, -NR ⁸ Or is absent;

n2 is 0, 1, 2 or 3;

Y ² is O, -NR ⁹ Or is absent;

R ⁸ and R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 0, 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

L ^a 、L ^b and L ^c Is a linking group;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand;

scheme 2:

R ^1a-1 independently is hydroxy, halogen or C ₁ -C ₁₂ An alkyl group;

R ^2a 、R ^4a 、R ^2b 、R ^4b 、R ^2c and R ^5c Independently H or halogen;

R ^4c is H, 3-6 membered heterocycloalkyl or is-N (R) ^4c-1 R ^4c-2 ) Substituted 3-6 membered heterocycloalkyl, the heteroatom in said 3-6 membered heterocycloalkyl and the nitrogen atom in said 3-6 membered heterocycloalkyl ^4c-1 R ^4c-2 ) The heteroatom in the substituted 3-6 membered heterocycloalkyl is independently one or more of N, O or S, and the number of the heteroatoms is 1-3;

R ^4c-1 And R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl, -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) — or absent; said 3-9 membered heterocycloalkyl and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 The heteroatom in the' is one or more of N, O or S independently, and the number of the heteroatom is 1-3;

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ An alkyl group;

Y ¹ is O, -NR ⁸ Or is absent (i.e. M) ¹ And L ^a Directly connected);

n2 is 0, 1, 2 or 3;

R ⁸ and R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 0, 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

L ^a 、L ^b and L ^c Connecting groups for connection;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand;

scheme 3:

R ^1a-1 is hydroxy, halogen or C ₁ -C ₁₂ An alkyl group;

R ^4c-1 and R ^4c-2 Independently is C ₁ -C ₄ An alkyl group;

X ^a is composed of

Y ¹ And L ^a Are connected to each other, Y ² And L ^b Connecting;

n1 is 1, 2 or 3;

M ¹ is 3-9 membered heterocycloalkyl, substituted by M ^1-1 Substituted 3-9 membered heterocycloalkyl, -3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 ' OC (= O) — or absent; said 3-9 membered heterocycloalkyl group and said 3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 The heteroatom in the' is one or more of N, O or S independently, and the number of the heteroatom is 1 to 3;

n1' is 1, 2 or 3;

M ^1-1 is C ₁ -C ₄ An alkyl group;

Y ¹ is O, -NR ⁸ Or is absent;

n2 is 0, 1, 2 or 3;

Y ² is O, -NR ⁹ Or is absent；

R ⁸ And R ⁹ Independently is C ₁ -C ₄ An alkyl group;

X ^b is composed of

n3 is 1, 2 or 3;

n3' is 1, 2 or 3;

R ¹⁰ and R ¹¹ Independently is C ₁ -C ₄ An alkyl group;

L ^a 、L ^b and L ^c Connecting the connecting ball;

Q ^a 、Q ^b and Q ^c Is an E3 ubiquitin ligase ligand;

scheme 4:

the general formula of the compound shown in the formula Ia is shown in formula Ia-1:

wherein R is ^2a Is halogen;

R ^5a independently is halogen;

X ^a is composed of

M ¹ Is-3-9 membered heterocycloalkyl- (CH) ₂ ) _n1 OC (= O) -or absent, Y ¹ is-NR ⁸ (ii) a Or, M ¹ Is 3-9 membered heterocycloalkyl or by M ^1-1 Substituted 3-9 membered heterocycloalkyl, Y ¹ Is O or absent;

L ^a is composed of

Scheme 5:

wherein R is ^2a Is halogen;

R ^5a Is halogen;

X ^a is composed of

L ^a Is composed of

Scheme 6:

the general formula of the compound shown in the formula Ia is Ia-2:

wherein R is ^2a Is halogen; r ^5a Is halogen;

scheme 7:

the general formula of the compound shown in the formula Ia is shown in formula Ia-3:

wherein R is ^2a Is halogen;

R ^5a is halogen;

X ^a is composed of

Y ¹ is absent;

L ^a is composed of

Scheme 8:

the general formula of the compound shown in the formula Ib is shown in a formula Ib-1:

wherein R is ^5a Is halogen;

R ^2b is halogen;

X ^b is composed of

Scheme 9:

the general formula of the compound shown in the formula Ib is shown in formula Ib-1:

wherein R is ^5a Is halogen;

R ^2b is halogen;

X ^b is composed of

n3' is 0;

L ^b is composed of

n7 is 0, n8 is 3, 4 or 5 ³ Is CH ₂ Or is absent;

n10 and n13 are 0, n11 and n12 are independently 3, 4 or 5 ⁴ Is absent;

scheme 10:

the general formula of the compound shown in the formula Ic is formula Ic' or Ic ":

preferably, in said formula Ic' or Ic ":

R ^2a and R ^5c Independently is halogen;

L ^c is composed of

n4 and n6 are 2, n5 is 4;

8. The compound of formula Ia, ib, or Ic, or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, metabolite, or isotopic compound thereof according to claim 1, wherein the compound of formula Ia, ib, or Ic is any one of:

For example

9. The compound of formula Ia, ib or Ic, or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, metabolite or isotopic compound thereof according to claim 1, wherein the pharmaceutically acceptable salt of the compound of formula Ia, ib or Ic is a formate, trifluoroacetate or hydrochloride;

and/or the number of the salts in the pharmaceutically acceptable salts of the compounds shown in the formulas Ia, ib or Ic is 1, 2, 3, 4, 5 or 6.

10. The compound of formula Ia, ib, or Ic, or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, metabolite, or isotopic compound thereof of claim 1, wherein the pharmaceutically acceptable salt of the compound of formula Ia, ib, or Ic is any one of the following compounds:

for example

11. The compound of formula Ia, ib, or Ic, or a pharmaceutically acceptable salt, solvate, stereoisomer, tautomer, prodrug, metabolite, or isotopic compound thereof according to claim 1, wherein the compound of formula Ia, ib, or Ic is any one of:

The pharmaceutically acceptable salt of the compound shown in the formula Ia, ib or Ic is any one of the following compounds:

12. a compound of formula IIa or IIb:

wherein R is ^12a And R ^12b Independently a hydroxyl protecting group; r ^13a And R ^13b Independently an amino protecting group;

R ^2a 、R ^4a 、R ⁶ 、R ⁷ 、R ^12a 、R ^13a 、A ^a 、X ^a 、L ^a 、Q ^a 、R ^2b 、R ^4b 、R ^12b 、R ^13b 、A ^b 、X ^b 、L ^b 、Q ^b are as defined in any one of claims 1 to 10;

preferably, the compound of formula IIa or IIb has the formula:

more preferably, the compound shown in formula IIa or IIb is any one of the following compounds:

for example

13. A pharmaceutical composition comprising substance X and one or more pharmaceutical excipients, wherein substance X is a compound of formula Ia, ib or Ic according to any one of claims 1 to 11, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof or an isotopic compound thereof.

14. Use of a substance Y for the manufacture of a medicament for the treatment or prevention of a cancer mediated by KRAS mutations; the substance Y is a compound represented by formula Ia, ib or Ic, or a pharmaceutically acceptable salt, solvate, prodrug, metabolite or isotopic compound thereof according to any one of claims 1 to 11, or a pharmaceutical composition according to claim 13.

15. Use of a substance Y for the manufacture of a medicament for the treatment or prevention of cancer; substance Y is a compound of formula Ia, ib or Ic according to any one of claims 1-11, a pharmaceutically acceptable salt thereof, a solvate thereof, a stereoisomer thereof, a tautomer thereof, a prodrug thereof, a metabolite thereof or an isotopic compound thereof, or a pharmaceutical composition according to claim 13; the cancer is hematological cancer, pancreatic cancer, MYH-related polyposis, colorectal cancer or lung cancer.