CN117751108A - Pyridazinone compounds - Google Patents

Abstract

A series of pyridazinone compounds and pharmaceutically acceptable salts thereof, and application thereof in preparing medicaments for treating related diseases, and particularly discloses a compound shown in a formula (IV) and pharmaceutically acceptable salts thereof.

Description

Pyridazinone compounds

The present application claims priority as follows:

CN202110827385.9, application date 2021, month 07, 21;

CN202110872908.1, application date 2021, month 07, 30;

CN202111509969.8, application day 2021, 12 and 10;

CN202210399423.X, application day 2022, 04, 15;

CN202210730508.1, application date 2022, month 06 and 24.

Technical Field

The invention discloses a series of pyridazinone compounds and application thereof in preparing medicaments for treating related diseases.

Background

Adenosine diphosphate ribosylation (ADP-ribosylation) is a reversible post-transcriptional modification process of proteins, and is involved in regulating various biological processes in vivo, including transcription regulation, mRNA stability, cell division, protein degradation, etc., and plays an important role in maintaining gene stability, apoptosis, etc. The family of poly (adenosine diphosphate) ribose polymerases (PARPs), also known as adenosine diphosphate ribose transferases (ART), is the most studied adenosine diphosphate ribosyl transferase at present. It has now been found that humans express 17 PARP family proteins, PARP1-17. PARP family members can be divided into three different types, depending on their structure and function: polyprps capable of transferring multiple ADP-ribose, monoprps capable of transferring only one single ADP-ribose and lack of activity.

PARP7 belongs to the family of monoprops, and the substrates of PARP7 have been identified to comprise Aryl hydrocarbon receptor (AhR) and TANK Binding Kinase (TBK 1); PARP7 inhibits these substrate activities by ADP ribosylation. Studies have shown that PARP7 is highly expressed in many cancers, and survival of many cancer cells is dependent on PARP7, and inhibition of PARP7 can inhibit growth of cancer cells. On the other hand, PARP7 is involved in immune escape of cancer cells, and inhibition of PARP7 can reactivate TBK1 and restore intracellular nucleic acid induction pathway activity, thereby inducing expression and synthesis of type I IFN. Elevated type I IFN levels can prevent tumor cell proliferation and induce a stronger anti-tumor immune response.

RBN-2397 developed by Ribon company is a PARP7 small molecule inhibitor in clinical phase I at present, and preclinical research shows that RBN-2397 has obvious inhibition on proliferation of various tumor cells, and meanwhile, has definite drug effect in a mouse immune model.

In conclusion, small molecule inhibitors targeting PARP7 could be an advanced cancer treatment regimen.

Disclosure of Invention

The present invention provides a compound of formula (IV) or a pharmaceutically acceptable salt thereof

Wherein,

ring A is selected from 4-10 membered heterocycloalkyl and C _4-10 Cycloalkyl, the 4-10 membered heterocycloalkyl and C _4-10 Cycloalkyl groups are each independently optionally substituted with 1, 2 or 3R _a Substitution;

R _a are each independently selected from H, CN, OH, F, cl, br, -COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy, said-COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3R; ring B is selected from 5-6 membered heteroaryl;

L ₅ selected from single bonds, -O-, -S-, and-NR ₁ -；

R ₁ Are independently selected from H, D, -C _1-3 alkyl-OH, -C _1-3 alkyl-NH ₂ And C _1-3 Alkyl, said-C _1-3 alkyl-OH, -C _1-3 alkyl-NH ₂ And C _1-3 Alkyl is optionally substituted with 1, 2 or 3R _b Substitution;

R _b are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1,2 or 3R substitutions;

L ₂ and L ₄ Are independently selected from single bond, -CH ₂ -、-CH ₂ -CH ₂ -and-CH ₂ -CH ₂ -CH ₂ -, the-CH ₂ -、-CH ₂ -CH ₂ -and-CH ₂ -CH ₂ -CH ₂ -each independently optionally being substituted with 1 or 2R _c Substitution;

R _c are respectively and independently selected from H, F, cl, br, D, C _1-3 Alkyl, -O-C _1-3 Alkyl and-C _1-3 alkyl-O-C _1-3 Alkyl, said C _1-3 Alkyl is each independently optionally substituted with 1, 2 or 3R;

R _2a and R is _2b Are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

L ₁ selected from single bond, -CH ₂ -、-O-、-C(＝O)-、-C(＝O)-NR ₄ -and-NR ₄ -；

L ₃ Selected from single bond, -CH ₂ -、-O-、-C(＝O)-、-C(＝O)-NR ₄ -and-NR ₄ -；

R ₄ Selected from H, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

R ₃ are each independently selected from H, CN, OH, F, cl, br, -COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino, C _1-3 Alkoxy and C _3-6 Cycloalkyl, said-COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino, C _1-3 Alkoxy and C _3-6 Cycloalkyl groups are each independently optionally substituted with 1, 2 or 3R;

each R is independently selected from D, OH, CN, F, cl and Br;

n is selected from 1, 2 and 3;

the 4-10 membered heterocycloalkyl groups each independently contain 1, 2 or 3 heteroatoms or groups of heteroatoms selected from N, O, S and NH.

In some embodiments of the invention, the above-mentioned compounds are selected from

Wherein,

ring A is selected from 4-10 membered heterocycloalkyl and C _4-10 Cycloalkyl, the 4-10 membered heterocycloalkyl and C _4-10 Cycloalkyl groups are each independently optionally substituted with 1, 2 or 3R _a Substitution;

R _a are each independently selected from H, CN, OH, F, cl, br, -COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy, said-COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3R; ring B is selected from 5-6 membered heteroaryl;

L ₅ selected from single bonds, -O-, -S-, and-NR ₁ -；

R ₁ Are independently selected from H, D, -C _1-3 alkyl-OH, -C _1-3 alkyl-NH ₂ And C _1-3 Alkyl, said-C _1-3 alkyl-OH, -C _1-3 alkyl-NH ₂ And C _1-3 Alkyl is optionally substituted with 1, 2 or 3R _b Substitution;

R _b are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

L ₂ and L ₄ Independently selected from a listBond, -CH ₂ -、-CH ₂ -CH ₂ -and-CH ₂ -CH ₂ -CH ₂ -, the-CH ₂ -each independently optionally being substituted with 1 or 2R _c Substitution;

R _c are respectively and independently selected from H, F, cl, br, D, C _1-3 Alkyl, -O-C _1-3 Alkyl and-C _1-3 alkyl-O-C _1-3 Alkyl, said C _1-3 Alkyl is each independently optionally substituted with 1, 2 or 3R;

R _2a and R is _2b Are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

L ₁ selected from single bond, -O-, -C (=O) -NH-, and-NR ₄ -；

L ₃ Selected from single bond, -O-, -C (=O) -NH-, and-NR ₄ -；

R ₄ Selected from H, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

R ₃ are each independently selected from H, CN, OH, F, cl, br, -COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy, said-COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3R; each R is independently selected from D, OH, CN, F, cl and Br;

n is selected from 1, 2 and 3;

the 4-10 membered heterocycloalkyl groups each independently contain 1, 2 or 3 heteroatoms or groups of heteroatoms selected from N, O, S and NH.

In some embodiments of the invention, the above-mentioned compounds are selected from

Wherein n, ring A, ring B, L ₁ 、L ₂ 、L ₃ 、L ₄ 、L ₅ 、R _2a 、R _2b And R is ₃ As defined herein.

In some embodiments of the invention, the above-mentioned compounds are selected from

Wherein n, ring A, ring B, L ₁ 、L ₂ 、L ₃ 、R _2a 、R _2b 、R ₃ As defined herein.

The carbon atoms with "×" are chiral carbon atoms, either in the form of (R) or (S) single enantiomers or enriched in one enantiomer.

In some embodiments of the invention, the above-mentioned compounds are selected from

Wherein n, ring A, ring B, L ₁ 、L ₂ 、L ₃ 、L ₄ 、L ₅ 、R _a 、R _2a 、R _2b And R is ₃ As defined in claim 1.

In some embodiments of the invention, the above-mentioned compounds are selected from

Wherein,

x is-O-, -CH ₂ -, -NH-or-N (CH) ₃ )-；

Y is N, Z is CH; or Y is CH, Z is N;

ring C is selected from 4-10 membered heterocycloalkyl, said 4-10 membered heterocycloalkyl optionally being interrupted by 1, 2 or 3R _a Substitution;

p is 1 or 2;

n, ring A, ring B, L ₁ 、L ₂ 、L ₃ 、R _2a 、R _2b 、Rc、R ₃ As defined herein;

the carbon atoms with "×" are chiral carbon atoms, either in the form of (R) or (S) single enantiomers or enriched in one enantiomer.

In some embodiments of the invention, the above-mentioned compounds are selected from

Wherein,

ring A is selected from 4-10 membered heterocycloalkyl and C _4-10 Cycloalkyl, the 4-10 membered heterocycloalkyl and C _4-10 Cycloalkyl groups are each independently optionally substituted with 1, 2 or 3R _a Substitution;

R _a are each independently selected from H, CN, OH, F, cl, br, -COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy, said-COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3R; ring B is selected from 5-6 membered heteroaryl;

L ₅ selected from single bonds, -O-, -S-, and-NR ₁ -；

R ₁ Are independently selected from H, D, -C _1-3 alkyl-OH, -C _1-3 alkyl-NH ₂ And C _1-3 Alkyl, said-C _1-3 alkyl-OH, -C _1-3 alkyl-NH ₂ And C _1-3 Alkyl is optionally substituted with 1, 2 or 3R _b Substitution;

R _b are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

L ₂ and L ₄ Are independently selected from single bond, -CH ₂ -、-CH ₂ -CH ₂ -and-CH ₂ -CH ₂ -CH ₂ -, the-CH ₂ -each independently optionally being substituted with 1 or 2R _c Substitution;

R _c are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

R _2a and R is _2b Are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

L ₁ selected from single bonds, -O-, -C (=O) -and-NR ₄ -；

L ₃ Selected from single bonds, -O-, -C (=O) -and-NR ₄ -；

R ₄ Selected from H, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

R ₃ are each independently selected from H, CN, OH, F, cl, br, -COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy, said-COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3R; each R is independently selected from D, OH, CN, F, cl and Br;

the 4-10 membered heterocycloalkyl groups each independently contain 1, 2 or 3 heteroatoms or groups of heteroatoms selected from N, O, S and NH.

In some embodiments of the invention, the compound described above, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from

Wherein, ring A, ring B, L ₁ 、L ₂ 、L ₃ 、L ₄ 、L ₅ 、R _2a 、R _2b And R is ₃ As defined herein.

In some embodiments of the invention, the compound described above, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from

Wherein, ring A, ring B, L ₁ 、L ₂ 、L ₃ 、R _2a 、R _2b 、R ₃ As defined herein;

the carbon atoms with "×" are chiral carbon atoms, either in the form of (R) or (S) single enantiomers or enriched in one enantiomer.

In some embodiments of the invention, the ring A is selected from the group consisting of azetidinyl, piperidinyl, piperazinyl, 1, 4-diazacyclohexenyl, 3, 8-diazabicyclo [3.2.1 ]]Octyl and 2, 5-diazabicyclo [2.2.2]Octyl said azetidinyl, piperidinyl, piperazinyl, 1, 4-diazacyclohexenyl, 3, 8-diazabicyclo [3.2.1 ]Octyl and 2, 5-diazabicyclo [2.2.2]Octyl radicals are each independently optionally substituted with 1, 2 or 3R _a Instead, the other variables are as defined herein.

In some embodiments of the invention, the ring A is selected from the group consisting of piperidinyl, piperazinyl, 1, 4-diazacyclohexanyl, 3, 8-diazabicyclo [3.2.1] octanyl, and 2, 5-diazabicyclo [2.2.2] octanyl, with the other variables being as defined herein.

In some embodiments of the invention, ring B is selected from pyrimidinyl, pyridinyl, pyridazinyl, pyrazolyl, thiophenyl, and thiazolyl, and the other variables are as defined herein.

In some embodiments of the invention, ring B is selected from pyrimidinyl, pyridinyl, pyridazinyl, pyrazolyl, and thiazolyl, and the other variables are as defined herein.

In some embodiments of the invention, the compounds described above, or pharmaceutically acceptable salts thereof, wherein ring B is selected from pyrimidinyl and thiazolyl, and the other variables are as defined herein.

In some aspects of the invention, L as described above ₅ Selected from-NH-, and other variables are as defined herein.

In some aspects of the invention, L as described above ₄ Selected from-CH (CH) ₃ )-、-CH(CH ₂ OCH ₃ )-、-CH(CH ₃ )CH ₂ CH ₂ -、-CH(CH ₃ )CH ₂ -、-CH(CH ₂ CH ₂ CH ₃ )-、-CH(CH ₂ CH ₂ OCH ₃ ) -and-CH (CH) ₂ OH) -, and other variables are as defined herein.

In some aspects of the invention, L as described above ₄ Selected from-CH (CH) ₃ )-、-CH(CH ₂ OCH ₃ ) -and-CH (CH) ₂ OH) -, and other variables are as defined herein. In some embodiments of the invention, the above compound or a pharmaceutically acceptable salt thereof, wherein L ₄ Selected from-CH (CH) ₃ ) -, the other variables are as defined herein.

In some aspects of the invention, R is as defined above _2a And R is _2b Independently selected from H and F, and the other variables are as defined herein.

In some aspects of the invention, L as described above ₁ selecting-C (=O) -CH ₂ -, -C (=O) -NH-and-C (=O) -N (CH) ₃ ) -, the other variables are as defined herein.

In some aspects of the invention, L as described above ₁ selecting-C (=O) -CH ₂ -and-C (=o) -NH-, the other variables being as defined herein.

In some embodiments of the invention, the above compound or a pharmaceutically acceptable salt thereof, wherein L ₁ select-C (=o) and the other variables are as defined herein.

In some aspects of the invention, L as described above ₃ Selected from single bond, -CH ₂ -, -O-, -NH-and-N (CH) ₃ ) -, the other variables are as defined herein.

In some aspects of the invention, L as described above ₃ Selected from the group consisting of-O-, -NH-and-N (CH) ₃ ) -, the other variables are as defined herein. In some embodiments of the invention, the above compound or a pharmaceutically acceptable salt thereof, wherein L ₃ Selected from-O-and-NH-, and the other variables are as defined herein.

In some aspects of the invention, L as described above ₂ Selected from-CH ₂ -and single bonds, the other variables being as defined herein.

In some embodiments of the invention, the above compound or a pharmaceutically acceptable salt thereof, wherein L ₂ Selected from-CH ₂ -, the other variables are as defined herein.

In some aspects of the invention, the structural units described aboveSelected from the group consisting of The other variables are as defined herein.

In some aspects of the invention, the structural units described aboveSelected from the group consisting of The other variables are as defined herein. In some embodiments of the invention, the above-described compounds, or pharmaceutically acceptable salts thereof, wherein the structural unitsSelected from the group consisting ofThe other variables are as defined herein.

In some aspects of the invention, the structural units described aboveSelected from the group consisting of The other variables are as defined herein.

In some aspects of the invention, the structural units described aboveSelected from the group consisting of The other variables are as defined herein.

In some embodiments of the invention, the above-described compounds, or pharmaceutically acceptable salts thereof, wherein the structural unitsSelected from the group consisting of The other variables are as defined herein.

The present invention provides a compound of the formula:

In some embodiments of the invention, the above-mentioned compounds are selected from

Still other embodiments of the present invention are derived from any combination of the variables described above.

The invention also provides application of the compound or pharmaceutically acceptable salt thereof in preparing PARP7 inhibitor.

Technical effects

The present invention provides a novel PARP7 inhibitor. The molecule of the invention has better in vitro inhibition activity and selectivity to PARP7, and is expected to provide a new and more effective scheme for treating cancers.

Definition and description

The following terms and phrases used herein are intended to have the following meanings unless otherwise indicated. A particular term or phrase, unless otherwise specifically defined, should not be construed as being ambiguous or otherwise clear, but rather should be construed in a generic sense. When trade names are presented herein, it is intended to refer to their corresponding commercial products or active ingredients thereof.

The term "pharmaceutically acceptable" as used herein is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention prepared from the compounds of the present invention which have the specified substituents found herein with relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting such compounds with a sufficient amount of base in pure solution or in a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, the acid addition salts may be obtained by contacting such compounds with a sufficient amount of acid in pure solution or in a suitable inert solvent. Certain specific compounds of the invention contain basic and acidic functionalities that can be converted to either base or acid addition salts.

Pharmaceutically acceptable salts of the invention can be synthesized from the parent compound containing an acid or base by conventional chemical methods. In general, the preparation of such salts is as follows: prepared via reaction of these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both.

The term "effective amount" or "therapeutically effective amount" refers to an amount that is non-toxic but which achieves the desired effect, unless otherwise specified. Determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, and also on the particular active substance, a suitable effective amount in an individual case can be determined by one skilled in the art according to routine experimentation.

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

Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of each other.

Unless otherwise indicated, the term "cis-trans isomer" or "geometric isomer" is caused by the inability of a double bond or a single bond of a ring-forming carbon atom to rotate freely.

Unless otherwise indicated, the term "diastereoisomer" refers to stereoisomers of a molecule having two or more chiral centers and having a non-mirror relationship between the molecules.

Unless otherwise indicated, "(+)" means dextrorotation, "(-)" means levorotatory, "(±)" means racemization.

Unless otherwise indicated, with solid wedge bondsAnd a wedge-shaped dotted bondRepresenting the absolute configuration of a solid centre by straight solid keysAnd straight dotted line keyRepresenting the relative configuration of the three-dimensional center by wavy linesSolid key representing wedge shapeOr wedge-shaped dotted bondOr by wave linesRepresenting straight solid keysOr straight dotted line key

Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to the fact that at room temperature, different functional group isomers are in dynamic equilibrium and are capable of rapid interconversion. If tautomers are possible (e.g., in solution), chemical equilibrium of the tautomers can be reached. For example, proton tautomers (also known as proton tautomers) (prototropic tautomer) include interconversions by proton transfer, such as keto-enol isomerisation and imine-enamine isomerisation. Valence isomer (valance tautomer) includes the interconversion by recombination of some of the bond-forming electrons. A specific example of where keto-enol tautomerization is the interconversion between two tautomers of pentane-2, 4-dione and 4-hydroxypent-3-en-2-one.

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

Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or enantiomers. For example, where one isomer or enantiomer is present in an amount of 90% and the other isomer or enantiomer is present in an amount of 10%, the isomer or enantiomer excess (ee value) is 80%.

Optically active (R) -and (S) -isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or derivatization with chiral auxiliary wherein the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereomeric resolution is carried out by conventional methods well known in the art, and then the pure enantiomer is recovered. Furthermore, separation of enantiomers and diastereomers is typically accomplished by the use of chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amine).

The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more of the atoms comprising the compounds. For example, compounds can be labeled with radioisotopes, such as tritium @, for example ³ H) Iodine-125% ¹²⁵ I) Or C-14% ¹⁴ C) A. The invention relates to a method for producing a fibre-reinforced plastic composite For example, deuterium can be substituted for hydrogen to form a deuterated drug, and the bond between deuterium and carbon is stronger than the bond between normal hydrogen and carbon, so that the deuterated drug has the advantages of reducing toxic and side effects, increasing the stability of the drug, enhancing the curative effect, prolonging the biological half-life of the drug and the like compared with the non-deuterated drug. All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.

The term "optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

The term "substituted" means that any one or more hydrogen atoms on a particular atom is substituted with a substituent, which may include deuterium and variants of hydrogen, provided that the valence of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =o), it means that two hydrogen atoms are substituted.

The term "optionally substituted" means that the substituents may or may not be substituted, and the types and numbers of substituents may be arbitrary on the basis that they can be chemically achieved unless otherwise specified.

When any variable (e.g., R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0 to 2R, the group may optionally be substituted with up to two R's, and R's in each case have independent options. Furthermore, combinations of substituents and/or variants thereof are only permissible if such combinations result in stable compounds.

When the number of one linking group is 0, such as- (CRR) ₀ -represents that the linking group is a single bond, -C ₀ alkyl-A means that the structure is actually-A.

When the number of a substituent is 0, this indicates that the substituent is absent, such as-A- (R) ₀ Indicating that the structure is actually-a.

When a substituent is absent, it is meant that the substituent is absent, e.g., X in A-X is absent, meaning that the structure is actually A.

When one of the variables is selected from a single bond, the two groups to which it is attached are indicated as being directly linked, e.g., when L in A-L-Z represents a single bond, it is indicated that the structure is actually A-Z.

When a substituent is absent, it is meant that the substituent is absent, e.g., X in A-X is absent, meaning that the structure is actually A.

When the listed substituents do not indicate which atom is attached to the substituted group, such substituents may be bonded through any atom thereof, for example, a pyridyl group may be attached to the substituted group as a substituent through any carbon atom on the pyridine ring.

Where a bond of a substituent may be cross-linked to more than one atom of a ring, such substituent may be bonded to any atom of the ring, e.g. a building blockIt means that the substituent R may be substituted at any position on the cyclohexyl or cyclohexadiene.

When the exemplified linking group does not indicate its linking direction, its linking direction is arbitrary, for example,the linking group L is-M-W-, in which case-M-W-may be a group formed by linking the rings A and B in the same direction as the reading order from left to rightThe ring A and the ring B may be connected in a direction opposite to the reading order from left to rightCombinations of such linking groups, substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, when a group has one or more bondable sites, any one or more of the sites of the group may be bonded to other groups by chemical bonds. When the connection mode of the chemical bond is not positioned and the H atoms exist in the connectable site, the number of the H atoms of the site can be correspondingly reduced to be changed into the corresponding valence group along with the number of the connected chemical bond when the chemical bond is connected. The chemical bond of the site and other groups can be a straight solid line bondStraight dotted line keyOr wave linesAnd (3) representing. For example-OCH ₃ The straight solid line bond in (a) represents the connection to other groups through the oxygen atom in the group;the straight dashed bonds in (a) represent the attachment to other groups through both ends of the nitrogen atom in the group;the wavy line means that the carbon atoms at positions 1 and 2 in the phenyl group are attached to other groups;it means that any of the ligatable sites on the piperidinyl group may be attached to other groups by 1 chemical bond, including at leastThese 4 connection modes, even though H atom is drawn on-N-, areStill includeThe group of this linkage is only when 1 chemical bond is linked, the H at this site will be correspondingly reduced by 1 to the corresponding monovalent piperidinyl group.

Unless otherwise specified, the term "C _1-3 For "alkyl groupsIn the expression linear or branched saturated hydrocarbon radicals consisting of 1 to 3 carbon atoms. The C is _1-3 Alkyl includes C _1-2 And C _2-3 Alkyl groups, etc.; it may be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). C (C) _1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

Unless otherwise specified, "C _4-8 Cycloalkyl "means a saturated cyclic hydrocarbon group consisting of 4 to 8 carbon atoms, which includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spiro, fused and bridged rings. The C is _4-8 Cycloalkyl includes C _4-5 、C _4-6 、C _4-7 、C _5-6 、C _5-7 、C _5-8、 C _{6- 7} 、C _6-8 、C _7-8 、C ₄ 、C ₅ 、C ₆ 、C ₇ 、C ₈ Cycloalkyl groups, and the like; it may be monovalent, divalent or multivalent. C (C) _4-8 Examples of cycloalkyl groups include, but are not limited to, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [ 2.2.2.2]Bicyclooctane, and the like.

Unless otherwise specified, the term "4-10 membered heterocycloalkyl" alone or in combination with other terms, respectively, denotes a saturated cyclic group consisting of 4 to 10 ring atoms, 1,2, 3 or 4 of which are heteroatoms independently selected from O, S and N, the remainder being carbon atoms, wherein the nitrogen atoms are optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms may optionally be oxidized (i.e. C (=o), NO and S (O) p, p being 1 or 2). It includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spiro, fused and bridged rings, as well as those wherein one ring is aromatic. In addition, in the case of the "4-8 membered heterocycloalkyl" group, the heteroatom may occupy the position of attachment of the heterocycloalkyl group to the remainder of the molecule. The 4-8 membered heterocycloalkyl group includes 4-6 membered, 4-7 membered, 4-8 membered, 5-6 membered, 5-7 membered, 5-8 membered, 6-7 membered, 6-8 membered, 7-8 membered, 4 membered, 5 membered, 6 membered, 7 membered, 8 membered, 9 membered, 10 membered heterocycloalkyl group and the like. Examples of 4-10 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl, 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl, 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1, 2-oxazinyl, 1, 2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, or dioxacycloheptyl, etc.

The terms "5-6 membered heteroaryl ring" and "5-6 membered heteroaryl" are used interchangeably herein unless otherwise specified, the term "5-6 membered heteroaryl" meaning a monocyclic group of 5 to 6 ring atoms having a conjugated pi-electron system, 1,2,3 or 4 of which are heteroatoms independently selected from O, S and N, the remainder being carbon atoms, wherein the nitrogen atoms are optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms are optionally oxidized (i.e., C (=o), NO and S (O) p, p being 1 or 2). The 5-6 membered heteroaryl group may be attached to the remainder of the molecule through a heteroatom or carbon atom. The 5-6 membered heteroaryl groups include 5-and 6-membered heteroaryl groups. Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl, 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, etc.), triazolyl (1H-1, 2, 3-triazolyl, 2H-1,2, 3-triazolyl, 1H-1,2, 4-triazolyl, 4H-1,2, 4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, etc.), furanyl (including 2-furanyl, 3-furanyl, etc.), thienyl (including 2-thienyl, 3-thienyl, etc.), pyridyl (including 2-pyridyl, 4-pyrimidyl, etc.), pyrimidyl (including 2-pyridyl, 4-pyrimidyl, etc.), pyrimidyl, etc.

Unless otherwise specified, "C _3-10 Cycloalkyl "means a group of 4 to 10 carbon atomsSaturated cyclic hydrocarbon groups are formed, which include monocyclic, bicyclic and tricyclic ring systems, wherein bicyclic and tricyclic ring systems include spiro, fused and bridged rings. The C is _3-10 Cycloalkyl includes C _4-8 、C _4-6 、C _4-5 、C _4-10 、C _4-6 、C _4-5 、C _5-8 Or C _5-6 Etc.; it may be monovalent, divalent or multivalent. C (C) _4-10 Examples of cycloalkyl groups include, but are not limited to, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [ 2.2.2.2]Bicyclo octane, [4.4.0 ]]Bicyclodecane, etc.

Unless otherwise specified, the term "C _1-3 Alkoxy "means those alkyl groups containing 1 to 3 carbon atoms that are attached to the remainder of the molecule through one oxygen atom. The C is _1-3 Alkoxy includes C _1-2 、C _2-3 、C ₃ And C ₂ Alkoxy groups, and the like. C (C) _1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

Unless otherwise specified, the term "C _1-3 Alkylamino "means those alkyl groups containing 1 to 3 carbon atoms attached to the remainder of the molecule through an amino group. The C is _1-3 Alkylamino includes C _1-2 、C ₃ And C ₂ Alkylamino, and the like. C (C) _1-3 Examples of alkylamino groups include, but are not limited to, -NHCH ₃ 、-N(CH ₃ ) ₂ 、-NHCH ₂ CH ₃ 、-N(CH ₃ )CH ₂ CH ₃ 、-NHCH ₂ CH ₂ CH ₃ 、-NHCH ₂ (CH ₃ ) ₂ Etc.

Unless otherwise specified, the term "C _1-3 Alkylthio "means those alkyl groups containing 1 to 3 carbon atoms which are attached to the remainder of the molecule through a sulfur atom. The C is _1-3 Alkylthio includes C _1-3 、C _1-2 And C ₃ Alkylthio, and the like. C (C) _1-3 Examples of alkylthio groups include, but are not limited to, -SCH ₃ 、-SCH ₂ CH ₃ 、-SCH ₂ CH ₂ CH ₃ 、-SCH ₂ (CH ₃ ) ₂ Etc.

Unless otherwise specified, the term "halo" or "halogen" by itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.

The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by combining with other chemical synthetic methods, and equivalent alternatives well known to those skilled in the art, preferred embodiments including but not limited to the examples of the present invention.

The compounds of the present invention may be structured by conventional methods well known to those skilled in the art, and if the present invention relates to the absolute configuration of a compound, the absolute configuration may be confirmed by conventional means in the art. For example, single crystal X-ray diffraction (SXRD), the grown single crystal is collected from diffraction intensity data using a Bruker D8 vent diffractometer, and the light source is cukα radiation, scanning:after scanning and collecting the relevant data, the absolute configuration can be confirmed by further analyzing the crystal structure by a direct method (Shellxs 97).

The solvent used in the present invention is commercially available.

The invention adopts the following abbreviations: aq represents water; HATU represents O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate; EDC represents N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride; m-CPBA represents 3-chloroperoxybenzoic acid; eq represents equivalent, equivalent; CDI represents carbonyldiimidazole; DCM represents dichloromethane; PE represents petroleum ether; DIAD represents azodicarboxylic acid diisoPropyl esters; DMF represents N, N-dimethylformamide; DMSO represents dimethylsulfoxide; etOAc represents ethyl acetate; etOH stands for ethanol; meOH represents methanol; CBz represents benzyloxycarbonyl, an amine protecting group; BOC represents that tert-butoxycarbonyl is an amine protecting group; HOAc stands for acetic acid; naCNBH ₃ Represents sodium cyanoborohydride; r.t. stands for room temperature; O/N stands for overnight; THF represents tetrahydrofuran; boc ₂ O represents di-tert-butyl dicarbonate; TFA represents trifluoroacetic acid; DIPEA stands for diisopropylethylamine; SOCl ₂ Represents thionyl chloride; CS (circuit switching) ₂ Represents carbon disulphide; tsOH represents p-toluenesulfonic acid; NFSI represents N-fluoro-N- (benzenesulfonyl) benzenesulfonamide; NCS represents 1-chloropyrrolidine-2, 5-dione; n-Bu ₄ NF represents tetrabutylammonium fluoride; iPrOH stands for 2-propanol; mp represents the melting point; LDA represents lithium diisopropylamide.

Compounds are either prepared according to the general nomenclature of the art or are usedSoftware naming, commercial compounds are referred to by vendor catalog names.

Detailed Description

The present invention is described in detail below by way of examples, but is not meant to be limiting in any way. The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by combining with other chemical synthetic methods, and equivalent alternatives well known to those skilled in the art, preferred embodiments including but not limited to the examples of the present invention. Various changes and modifications to the specific embodiments of the invention will be apparent to those skilled in the art without departing from the spirit and scope of the invention.

Example 1

Step A: sodium hydrogen (11.34 g, 283.60 mmol, 60% purity, 1.2 eq.) is slowly added in portions to a solution of compound 1-1 (60 g, 236.33 mmol, 1 eq.) in N, N-dimethylformamide (500 ml) at 0-10 degrees celsius, then p-methoxybenzyl chloride (44.41 g, 283.60 mmol, 38.62 ml, 1.2 eq.) is added and the mixture stirred at 25 degrees celsius for 6 hours. And (3) slowly adding a saturated ammonium chloride aqueous solution into the reaction solution under stirring until no gas is discharged, filtering the mixed solution, washing a filter cake with water (3 ml multiplied by 2), and drying the filter cake under reduced pressure to obtain the compound 1-2. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 8.15(s,1H),7.27-7.25(m,2H),6.90-6.87(m,2H),5.18(s,2H),3.72(s,3H)。MS(ESI)m/z:374.8[M+H ⁺ ]。

And (B) step (B): to a solution of compounds 1-2 (75 g, 200.52 mmol, 1 eq.) in methanol (750 ml) was added potassium hydroxide (33.75 g, 601.56 mmol, 3 eq.) and the mixture was stirred at 25 degrees celsius for 6 hours. The reaction solution was concentrated to 100 to 150 ml, and the above-mentioned liquid was filtered to collect a cake. The above cake was added to water (500 ml), stirred at 25 ℃ for 30 minutes, filtered, and the cake was dried under reduced pressure to give compounds 1-3. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 8.15(s,1H),7.24(d,J＝8.8Hz,2H),6.89-6.86(m,2H),5.21(s,2H),4.05(s,3H),3.72(s,3H)。

Step C: to N-methylpyrrolidone (500 ml) of compounds 1 to 3 (55 g, 169.15 mmol, 1 eq.) and compounds 1 to 4 (97.49 g, 507.45 mmol, 64.56 ml, 3 eq.) was added cuprous iodide (16.11 g, 84.57 mmol, 0.5 eq.) and the mixture was heated to 100 degrees celsius and stirred for 3 hours. After cooling to room temperature, water (150 ml) was added to the reaction solution, extracted with methylene chloride (200 ml. Times.3), the organic phases were combined, washed with saturated brine (200 ml. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a residue which was purified by column chromatography1000 gSilica gel column, eluent gradient 0-100% ethyl acetate/petroleum ether 66 ml/min) to obtain compound 1-5.MS (ESI) m/z 315.3[ M+H ] ⁺ ]。

Step D: to a solution of compounds 1-5 (31.11 g, 98.99 mmol, 1 eq.) in N, N-dimethylformamide (250 ml) was added trimethyliodosilane (29.71 g, 148.49 mmol, 20.21 ml, 1.5 eq.) and the mixture was heated to 85 degrees celsius and stirred for 12 hours. Cooling to room temperature, adding saturated ammonium chloride (150 ml) aqueous solution into the reaction solution, filtering the above mixture with diatomite, extracting the filtrate with ethyl acetate (200 ml×3), mixing the organic phases, washing with saturated saline (100 ml×2), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the obtained residue to column chromatography1000 gSilica gel column, eluent gradient 0-100% ethyl acetate/petroleum ether 44 ml/min) to obtain compound 1-6. ¹ H NMR(400MHz,DMSO-d ₆ )δppm7.78(s,1H),7.24(d,J＝8.8Hz,2H),6.89(d,J＝8.8Hz,2H),5.09(s,2H),3.72(s,3H)。MS(ESI)m/z:323.0,325.0[M+Na ⁺ ]。

Step E: oxalyl chloride (10.15 g, 79.94 mmol, 7.00 ml, 2 eq.) was added to a solution of compounds 1-6 (12 g, 39.97 mmol, 1 eq.) in N, N-dimethylformamide (120 ml) at 0 degrees celsius and the mixture stirred at 25 degrees celsius for 8 hours. The reaction mixture was quenched with water (200 ml), extracted with ethyl acetate (150 ml. Times.3), and the organic phases were combined and washed with saturated brine (100 ml. Times.2)Washing with water, drying with anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the residue to column chromatography 120 gSilica gel column, eluent gradient 0-100% ethyl acetate/petroleum ether 66 ml/min) to obtain compound 1-7. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 7.78(s,1H),7.24(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.19(s,2H),3.72(s,3H)。

Step F: to a solution of compounds 1-8 (4.3 g, 23.56 mmol, 1 eq.) in N-methylpyrrolidone (50 ml) under nitrogen protection, compounds 1-9 (4.39 g, 23.56 mmol, 1 eq.) and potassium carbonate (6.51 g, 47.12 mmol, 2 eq.) were added and the reaction stirred at 80℃for 1 hour. After the reaction solution was cooled to room temperature, water (200 ml) was added thereto, and a white solid was precipitated, filtered and the cake was collected, and dried under reduced pressure to obtain compounds 1 to 10.MS (ESI) m/z 277.1[ M+H ] ⁺ -56]。

Step G: to a solution of 1-10 (7.8 g, 23.47 mmol, 1 eq.) in anhydrous dioxane (80 ml) was added dioxane hydrochloride (4 mol per liter, 78.00 ml, 13.29 eq.) and the reaction stirred at 25 degrees celsius for 1 hour. Filtering at room temperature, collecting filter cake, and drying under reduced pressure to obtain hydrochloride of the compound 1-11. MS (ESI) m/z 233.1[ M+H ] ⁺ ]。

Step H: to a solution of compounds 1-12 (3 g, 17.32 mmol, 1 eq.) and compounds 1-13 (3.88 g, 17.32 mmol, 3.44 ml, 1 eq.) in tetrahydrofuran (10 ml) was added potassium tert-butoxide (2.92 g, 25.98 mmol, 1.5 eq.) and the reaction stirred at 25 degrees celsius for 12 hours. To the reaction solution was added water (100 ml), extracted with ethyl acetate (100 ml×2), and the organic phases were combined, washed with saturated brine (100 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1) to give compounds 1 to 14.MS (ESI) m/z 144.2[ M+H ] ⁺ -100]。

Step I: compounds 1-14 (900 mg, 3.70 mmol, 1 eq.) were dissolved in dioxane hydrochloride (10 ml) and stirred at 25℃for 15 min. The reaction solution was concentrated to obtain hydrochloride salts of the compounds 1 to 15. MS (ESI) m/z 144.1[ M+H ] ⁺ ]。

Step J: to a solution of compounds 1-15 (700 mg, 3.90 mmol, 1 eq, hydrochloride) in N, N-dimethylformamide (20 ml) was added triethylamine (1.18 g, 11.69 mmol, 1.63 ml, 3 eq) and compounds 1-7 (1.24 g, 3.90 mmol, 1 eq). The reaction solution was stirred at 25 degrees celsius for 12 hours. Water (60 ml) was added to the reaction solution, ethyl acetate (60 ml. Times.2) was extracted, the organic phases were combined, washed with saturated brine (60 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1 to 2/1) to give compounds 1 to 16.MS (ESI) m/z 426.3[ M+H ] ⁺ ]。

Step K: to a solution of compounds 1-16 (1 g, 2.35 mmol, 1 eq.) in trifluoroacetic acid (10 ml) was added trifluoromethanesulfonic acid (1.76 g, 11.75 mmol, 1.04 ml, 5 eq.) and the reaction was stirred at 25 degrees celsius for 1 hour. The reaction solution was poured into water (100 ml), pH was adjusted to 7 to 8 with sodium bicarbonate, ethyl acetate (100 ml×2) was extracted, the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1 to 1/1) to give compounds 1 to 17.MS (ESI) m/z 306.1[ M+H ] ⁺ ]。

Step L: to a solution of compounds 1-17 (460 mg, 1.51 mmol, 1 eq.) in tetrahydrofuran (10 ml) at 0 degrees celsius was added diisobutylaluminum hydride (1 mol per liter, 4.52 ml, 3 eq.) and the reaction was stirred for 30 minutes at 0 degrees celsius. To the reaction solution was added saturated aqueous sodium sulfate (5 ml), followed by ethyl acetate (100 ml), stirring was continued at 25 ℃ for 10 minutes, the above mixture was filtered, the cake was washed with ethyl acetate (30 ml), the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1 to 1/1), to obtain compounds 1 to 18.MS (ESI) m/z 264.2[ M+H ] ⁺ ]。

Step M: to a solution of compounds 1-18 (230 mg, 873.81 mmol, 1 eq.) in dichloromethane (5 ml) was added N, N-carbonyldiimidazole (170.03 mg, 1.05 mmol, 1.2 eq.) and the mixture was stirred at 25 ℃ for 30 min, then 1-11 (352.14 mg, 1.31 mmol, 1.5 eq.) and triethylamine (265.26 mg, 2.62 mmol, 364.87 μl, 3 eq.) in dichloromethane (5 ml) were added. The reaction solution was stirred at 25 degrees celsius for 12 hours. The reaction solution was poured into water (80 ml), extracted with ethyl acetate (80 ml. Times.2), the organic phases were combined, washed with saturated brine (80 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by high performance liquid chromatography (column: waters Xbridge 150 x 25 mm. Times.5 μm; mobile phase: [ water (10 mmol/l ammonium bicarbonate) -acetonitrile ]The method comprises the steps of carrying out a first treatment on the surface of the Ammonium bicarbonate%: 40% -70%,8 min) to obtain the compounds 1-19.MS (ESI) m/z 522.3[ M+H ] ⁺ ]。 ¹ H NMR (400 MHz, chloroform-d) delta ppm 10.22 (s, 1H), 8.51 (s, 2H), 7.58 (s, 1H), 5.84-5.75 (m, 2H), 5.47-5.44 (m, 1H), 4.67-4.62 (m, 2H), 4.29-4.26- (m, 1H), 3.93-3.91 (m, 4H), 3.58-3.56 (m, 4H), 1.46 (d, J=6.4 Hz, 3H).

Step N: compounds 1-19 (5.9 g, 11.32 mmol, 1 eq.) were isolated by chiral resolution (column: DAICEL CHIRALCEL OJ (250 mm. Times.30 mm, 10. Mu.m.) mobile phase: [0.1% aqueous ammonia methanol)]The method comprises the steps of carrying out a first treatment on the surface of the 25% -25% methanol for 3 min) to give compound 1a.MS (ESI) m/z 522.2[ M+H ] ⁺ ]。 ¹ H NMR (400 MHz, chloroform-d)δppm 10.91(s,1H),8.51(s,2H),7.59(s,1H),5.88-5.72(m,2H),5.54-5.39(m,1H),4.75-4.58(m,2H),4.35-4.24(m,1H),4.00–3.88(m,4H),3.70-3.52(m,4H),1.45(d,J＝6.8Hz,3H).

Compound 1a was tested by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 1a was 1.162min and the e.e. value was 100%.

Example 2

Step A: to a solution of compounds 1-17 (420 mg, 1.38 mmol, 1 eq.) in tetrahydrofuran (10 ml) at 0 degrees celsius was added diisobutylaluminum hydride (1 mol per liter, 4.13 ml, 3 eq.) and the reaction was stirred for 30 minutes at 0 degrees celsius. Saturated aqueous sodium sulfate (10 ml) was added to the reaction solution, followed by ethyl acetate (100 ml) and stirring was continued at 25 ℃ for 10 minutes, the above mixture was filtered, the cake was washed with ethyl acetate (100 ml), the filtrate was concentrated, and the crude product was purified by silica gel thin layer chromatography (petroleum ether/ethyl acetate=1/2) to give a product by chiral resolution (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm, 10 μm); mobile phase: [0.1% aqueous ammonia isopropanol ]The method comprises the steps of carrying out a first treatment on the surface of the Isopropyl alcohol%: 20% -20%,5.3 min) to give compounds 1-18a. MS (ESI) m/z 264.1[ M+H ] ⁺ ]。

Compounds 1-18a were tested by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is isopropanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compounds 1-18a was 1.146min and the e.e. value was 99.122%.

And (B) step (B): to a solution of compounds 1-18a (200 mg, 759.83 μmol, 1 eq.) in toluene (10 ml) was added 1, 8-diazacycloundec-7-ene (173.51 mg, 1.14 mmol, 171.79 μl, 1.5 eq.) and diphenyl azide phosphate (313.66 mg, 1.14 mmol, 246.98 μl, 1.5 eq.) and the mixture was stirred at 80 degrees celsius for 2 hours. The reaction mixture was concentrated and the crude product was purified by column chromatography (silica, petroleum ether/ethyl acetate=10/1, 1/1) to give compound 2-1.MS (ESI) m/z 289.1[ M+H ] ⁺ ].

Step C: triphenylphosphine (245.70 mg, 936.76 μmol, 1.5 eq.) was added to a solution of compound 2-1 (180 mg, 624.50 μmol, 1 eq.) in tetrahydrofuran (5 ml) and the mixture was stirred at 25 degrees celsius for 30 minutes. Water (0.5 ml) was further added to the reaction mixture, and the mixture was stirred at 40℃for 12 hours. To the reaction solution was added water (30 ml) for dilution, which was extracted with ethyl acetate (30 ml), and the aqueous phase was concentrated to give compound 2-2.MS (ESI) m/z 263.1[ M+H ] ⁺ ].

Step D: to a solution of compound 2-2 (120 mg, 457.61 μmol, 1 eq) in N, N-dimethylformamide (5 ml) was added N, N-carbonyldiimidazole (96.46 mg, 594.89 μmol, 1.3 eq) and diisopropylethylamine (295.71 mg, 2.29 mmol, 398.53 μmol, 5 eq) and the mixture was stirred at 25 degrees celsius for 2 hours. Compounds 1 to 11 (212.52 mg, 915.22. Mu. Mol, 2 eq.) were then added to the reaction mixture and the mixture was stirred at 60℃for 12 hours. The reaction solution was subjected to high performance liquid chromatography (column: unisil 3-100 C18 Ultra 150*50 mm. 3 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 39% -69%,7 min) to give compound 2.MS (ESI) m/z 521.3[ M+H ] ⁺ ]. ¹ H NMR (400 MHz, chloroform-d) delta ppm 11.05 (br s, 1H), 8.51 (s, 2H), 7.61 (s, 1H) 5.59-5.80 (m, 2H), 5.50-5.39 (m, 1H), 4.85-4.82 (m, 1H), 4.32-4.19 (m, 1H), 4.02-3.82 (m, 6H), 3.61-3.45 (m, 4H), 1.42 (d, J=6.8 Hz, 3H).

Compound 2 was tested by SFC [ column model: chiralpak AD-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 2 was 1.702min and the e.e. value was 99.390%.

Example 3

Step A: compounds 1-18 (516 mg, 1.96 mmol, 1 eq.) were dissolved in toluene (6 ml) and 1.8-diazabicyclo [5.4.0] undec-7-ene (358.13 mg, 2.35 mmol, 354.58. Mu.l, 1.2 eq.) and diphenyl azide phosphate (647.39 mg, 2.35 mmol, 509.76. Mu.l, 1.2 eq.) were added. The reaction solution was stirred at 80 degrees celsius for 2 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, diluted with water (50 ml), extracted with ethyl acetate (50 ml×3), and the combined organic phases were washed with saturated brine (50 ml×1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product by column chromatography (silica gel, petroleum ether/ethyl acetate=3/1 to 1/1) to give compound 3-1.

And (B) step (B): compound 3-1 (280 mg, 971.45 μmol, 1 eq) was dissolved in tetrahydrofuran (3 ml) and triphenylphosphine (382.20 mg, 1.46 mmol, 1.5 eq) was added. The reaction solution was stirred at 25 degrees celsius for 0.5 hours, and water (0.5 ml) was added to the reaction solution. The reaction solution was stirred at 25 degrees celsius for 1 hour. After completion of the reaction, ethyl acetate (50 ml) was added to dilute the mixture, and the mixture was washed with water (50 ml. Times.4). The combined aqueous phases are directly concentrated to obtain the compound 3-2.MS (ESI) m/z 263.0[ M+H ] ⁺ ]。

And (B) step (B): compound 1-8 (500 mg, 2.74 mmol, 1 eq.) was dissolved in N, N-dimethylformamide (5 ml), and compound 3-3 (430.64 mg, 2.74 mmol, 422.19 μl, 1 eq.) and potassium carbonate (757.16 mg, 5.48 mmol, 2 eq.) were added. The reaction solution was stirred at 90 degrees celsius for 1 hour. After completion of the reaction, the reaction mixture was cooled to room temperature, diluted with water (10 ml), and extracted with ethyl acetate (10 ml×3). The combined organic phases were washed with saturated brine (10 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compounds 3-4.

Step C: compound 3-4 (0.75 g, 2.47 mmol, 1 eq.) was dissolved in methanol (10 ml) and water (10 ml), lithium hydroxide monohydrate (311.32 mg, 7.42 mmol, 3 eq.) was added and the reaction stirred at 25 degrees celsius for 1 hour. After completion of the reaction, hydrochloric acid (1 mol/l) was added to the reaction mixture to adjust the pH to 5, extraction was performed with ethyl acetate (20 ml. Times.3), and the combined organic phases were washed with saturated brine (20 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give Compound 3-5.MS (ESI) m/z 276.0[ M+H ] ⁺ ].

Step D: compound 3-5 (83.96 mg, 305.07 μmol, 1 eq) was dissolved in tetrahydrofuran (1 ml), triethylamine (61.74 mg, 610.15 μmol, 84.93 μmol, 2 eq) and O- (7-azabenzotriazol-1-yl) N, N-tetramethylurea hexafluorophosphate (174.00 mg, 457.61 μmol, 1.5 eq) were added, and the reaction mixture was stirred at 25 degrees celsius for 30 minutes, and compound 3-2 (80 mg, 305.07 μmol, 1 eq) was added. The reaction solution was stirred at 25 degrees celsius for 1 hour. After completion of the reaction, the reaction mixture was diluted with water (50 ml), extracted with ethyl acetate (50 ml×3), and the combined organic phases were washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Separating the crude product by thin layer chromatography (silica gel, petroleum ether/ethyl acetate=0/1), subjecting the obtained compound to high performance liquid chromatography (column: phenomenex Synergi C, 150×25 mm, 10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the 36% -56% acetonitrile, 9 min) to give compound 3.MS (ESI) m/z 520.2[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 8.67(s,2H),8.02(t,J＝5.2Hz,1H),7.80(s,1H),6.47-6.38(m,1H),5.62-5.57(m,2H),4.72-4.65(m,2H),4.54-4.46(m,1H),3.65(br s,2H),3.09–2.98(m,2H),2.46-2.42(m,1H),1.80-1.71(m,2H),1.55-1.42(m,2H),1.29(d,J＝5.6Hz,3H).

Example 4

And (B) step (B): to a solution of compound 4-1 (100 mg, 471.06 micromolar, 1 eq.) and compound 1-8 (85.98 mg, 471.06 micromolar, 1 eq.) in N, N-dimethylformamide (5 ml) was added potassium carbonate (130.21 mg, 942.12 micromolar, 2 eq.) and the reaction was heated to 90 degrees celsius and stirred for 2 hours. The reaction solution was poured into water (50 ml), extracted with ethyl acetate (50 ml), and the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 4-2.MS (ESI) m/z 303.1[ M+H ] ⁺ -56]。

Step C: compound 4-2 (160 mg, 446.48 μmol, 1 eq.) was added to ethyl acetate hydrochloride (5 ml, 4 mol/l) and the reaction stirred at 25 degrees celsius for 15 minutes. The reaction solution was concentrated to obtain hydrochloride of Compound 4-3. MS (ESI) m/z 259.1[ M+H ] ⁺ ]。

Step D: to a solution of compound 1-18a (20 mg, 75.98. Mu. Mol, 1 eq.) in N, N-dimethylformamide (1 ml) were added N, N-carbonyldiimidazole (24.64 mg, 151.96. Mu. Mol, 2 eq.) and triethylamine (30.75 mg, 303.92. Mu. Mol, 42.30. Mu.l, 4 eq.) and, after stirring at 25℃for 2 hours, compound 4-3 (33.59 mg, 113.97. Mu. Mol, 1.5 eq., hydrochloride) was added, and the reaction mixture was stirred at 25℃for 12 hours and then heated to 50℃and stirred for 24 hours. The reaction solution was poured into water (30 ml), extracted with ethyl acetate (30 ml. Times.2), the organic phases were combined, washed with saturated brine (30 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a solution prepared by HPLC (column: phenomenex luna C: 150 x 25 mm. Times.10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]: acetonitrile%: 41% -71%,10 min) to give compound 4.MS (ESI) m/z 550.2[ M+H ] ⁺ ]。 ¹ HNMR(400MHz,DMSO-d ₆ )δppm 12.52(s,1H),8.72(s,2H),7.81(s,1H),6.50-6.47(m,1H),5.65-5.90(m,2H),4.79(br s,2H),4.45-4.66(m,3H),3.73-3.88(m,2H),2.97-3.18(m,2H),1.87-1.98(m,2H),1.68-1.67(m,2H),1.31(d,J＝6.80Hz,3H).

Compound 4 was tested by SFC [ column model: chiralpak AD-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 4 was 1.853min and the e.e. value was 100%.

Example 5

Step A: to a solution of compound 5-1 (100 mg, 471.06 micromolar, 1 eq.) in N-methylpyrrolidone (2 ml) was added potassium carbonate (130.21 mg, 942.12 micromolar, 2 eq.) and compound 1-8 (85.98 mg, 471.06 micromolar, 1 eq.) and the mixture was stirred at 80 degrees celsius for 2 hours. After the reaction solution was cooled to 25 ℃, water (10 ml) was added, a yellow solid was precipitated, and the filter cake was filtered and collected, and dried under reduced pressure to obtain compound 5-2.MS (ESI) m/z 303.0[ M+H ] ⁺ -56].

And (B) step (B): to a solution of 5-2 (110 mg, 306.96 μmol, 1 eq.) in ethyl acetate (1 ml) was added ethyl acetate hydrochloride (4 mol, 1 ml, 13.03 eq.) and the mixture was stirred at 25 degrees celsius for 0.5 hours. The reaction solution was directly dried under reduced pressure to obtain hydrochloride of the compound 5-3. MS (ESI) m/z 259.2[ M+H ] ⁺ ].

Step C: to a solution of compounds 1-18a (30 mg, 113.97. Mu. Mol, 1 eq.) in N, N-dimethylformamide (2 ml) were added N, N-carbonyldiimidazole (36.96 mg, 227.95. Mu. Mol, 2 eq.) and triethylamine (34.60 mg, 341.92. Mu. Mol, 47.59. Mu.l, 3 eq.) and the mixture was stirred at 25℃for 12 hours, then compounds 5-3 (44.15 mg, 170.96. Mu. Mol, 1.5 eq.) were added and the mixture was stirred at 50℃24 hours. Diluting with water (10 ml), extracting with ethyl acetate (10 ml. Times.2), mixing the organic phases, washing with saturated saline (10 ml. Times.2), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to high performance liquid chromatography (column: phenomenex Synergi C, 150X 25 mm. Times.10 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 41% -71%,10 min) to give compound 5.MS (ESI) m/z 548.2[ M+H ] ⁺ ].1H NMR(400MHz,DMSO-d ₆ )δppm 12.51(br s,1H),8.70-8.69(m,2H),7.82-7.81(m,1H),6.49-6.48(m,1H),5.81-5.76(m,2H),4.98-4.96(m,1H),4.56-4.52(m,3H),4.31-4.28(m,1H),3.67(br s,2H),3.56-3.50(m,2H),1.90-1.78(m,4H),1.32-1.28(m,3H).

Compound 5 was detected by SFC [ column model: chiralpak AD-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 40% -40%) to obtain: the retention times for compound 5 were 0.651min and 1.124min, and the e.e. value was 1.394%.

Example 6

Step A: compound 6-1 (1.68 g, 6.93 mmol, 1.41 ml, 1.2 eq.) was dissolved in tetrahydrofuran (10 ml), and potassium hydroxide (323.92 mg, 5.77 mmol, 1 eq.) and compound 1-12 (1 g, 5.77 mmol, 1 eq.) were added. The reaction mixture was stirred at 25 degrees celsius for 12 hours. After the reaction was completed, water (50 ml) was added to the reaction solution, extraction was performed with ethyl acetate (50 ml. Times.3), the combined organic phase was washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was subjected to column chromatography25g Silica gel column, eluent gradient 0-6% petroleum ether: ethyl acetate 80 ml/min) to give compound 6-2.MS (ESI) m/z:206.0[ M+H ] ⁺ -56]. ¹ H NMR (400 MHz, chloroform-d) delta ppm 6.08-5.90 (m, 1H) 5.06-4.93 (m, 1H) 4.42-4.50 (m, 1H) 4.29-4.23 (m, 2H) 1.37 (s, 9H) 1.31-1.28 (m, 3H) 1-25-1.23 (m, 3H)

And (B) step (B): compound 6-2 (840 mg, 3.21 mmol, 1 eq) was dissolved in ethyl acetate (8 ml) and ethyl acetate hydrochloride (4 mol, 8 ml, 9.95 eq) was added. The reaction mixture was stirred at 25 degrees celsius for 1 hour. After the reaction was completed, the reaction solution was concentrated in vacuo to give hydrochloride of Compound 6-3. MS (ESI) m/z:162.1[ M+H ] ⁺ ]

Step C: compound 6-3 (724 mg, 4.49 mmol, 1 eq, hydrochloride) was dissolved in N, N-dimethylformamide (10 ml), and triethylamine (1.36 g, 13.48 mmol, 1.88 ml, 3 eq) and compound 1-7 (1.43 g, 4.49 mmol, 1 eq) were added. The reaction solution was stirred at 25 degrees celsius for 12 hours. After the reaction was completed, ethyl acetate (50 ml) was added to the reaction mixture, which was washed with water (50 ml. Times.3) and saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was subjected to column chromatography25g Silica gel column, eluent gradient 0-6% ethyl acetate: petroleum ether 80 ml/min) to give compound 6-4.MS (ESI) m/z:444.2[ M+H ] ⁺ ]

Step D: compound 6-4 (1.24 g, 2.80 mmol, 1 eq.) was dissolved in trifluoroacetic acid (10 ml) and trifluoromethanesulfonic acid (1.70 g, 11.33 mmol, 1 ml, 4.05 eq.) was added and the reaction mixture stirred at 25 degrees celsius for 10 minutes. After the reaction is completed, the reaction solution is slowly dropped into the saturated sodium carbonate solutionTo (200 ml) was added sodium carbonate solid to adjust pH to 9, followed by extraction with ethyl acetate (200 ml×3). The combined organic phases were washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was subjected to column chromatography 12g Silica gel column, eluent gradient 0-16% ethyl acetate: petroleum ether (1% methylene chloride) 60 ml/min) gives compound 6-5.MS (ESI) m/z:324.0[ M+H ] ⁺ ]

Step E: compound 6-5 (0.5 g, 1.55 mmol, 1 eq.) was dissolved in tetrahydrofuran (6 ml) and diisobutylaluminum hydride (1 mol, 9.28 ml, 6 eq.) was added at 0 degrees celsius. The reaction mixture was stirred at 25 degrees celsius for 1 hour. After completion of the reaction, a saturated ammonium chloride solution (5 ml) was added thereto to quench the reaction, and ethyl acetate (5 ml. Times.4) was extracted. The combined organic phases were washed with saturated brine (5 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give crude product which was plate separated (petroleum ether/ethyl acetate=1/2) to give the product which was then subjected to chiral resolution (column: DAICEL CHIRALCEL OJ (250 mm. Times.30 mm, 10 μm); mobile phase: [0.1% aqueous ammonia isopropanol]The method comprises the steps of carrying out a first treatment on the surface of the Isopropyl alcohol%: 15% -15%,5.6 minutes) to give compound 6-6.MS (ESI) m/z:282.0[ M+H ] ⁺ ]

Step F: compound 6-6 (92 mg, 327.16 μmol, 1 eq) was dissolved in tetrahydrofuran (4 ml), N-carbonyldiimidazole (106.10 mg, 654.32 μmol, 2 eq) and triethylamine (99.32 mg, 981.49 μmol, 136.61 μmol, 3 eq) were added, and the reaction mixture was stirred at 25 degrees celsius for 1 hour, and compound 1-11 (87.90 mg, 327.16 μmol, 1 eq, hydrochloride) was added to the reaction solution. The reaction mixture was stirred at 50 degrees celsius for 17 hours. After the reaction was completed, the reaction solution was cooled to 25℃and water (20 ml) and acetic acid were added to the reaction solution Ethyl ester (20 ml×4) extraction. The combined organic phases were washed with saturated brine (20 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the crude product which was subjected to high performance liquid chromatography (column: phenomenex Synergi C: 150 x 25 mm x 10 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 42% -72%,10 min) to give compound 6.MS (ESI) m/z:540.2[ M+H ] ⁺ ] ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.60(s,1H),8.73(s,2H),7.77(s,1H),6.66-6.58(m,1H),5.39(dd,J＝36.0Hz,1H),4.89-4.77(m,1H),4.69(d,J＝16.0Hz,2H),3.89-3.80(m,4H),3.52-3.44(m,4H),1.34(d,J＝8.0Hz,3H).

Compound 6 was tested by SFC [ column model: OD 50X 4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 6 was 1.189min and the e.e. value was 100%.

Example 7

Step A: compound 7-1 (0.5 g, 3.58 mmol, 1 eq.) and compound 1-9 (667.36 mg, 3.58 mmol, 1 eq.) were dissolved in N-methylpyrrolidone (10 ml) and potassium carbonate (990.45 mg, 7.17 mmol, 2 eq.) was added. The reaction mixture was stirred at 100 degrees celsius for 2 hours. After the reaction was completed, the reaction solution was cooled to 25 degrees celsius, the reaction solution was poured into water (30 ml), filtered, and the filter cake was spin-dried under vacuum to obtain compound 7-2. ¹ H NMR (400 MHz, chloroform-d) delta ppm 8.50-8.47 (m, 2H), 3.90-3.51 (m, 8H), 1.50 (s, 9H).

And (B) step (B): compound 7-2 (400 mg, 1.38 mmol, 1 eq.) was dissolved in ethyl acetate (2 ml) and ethyl acetate hydrochloride (4 mol per liter, 2 ml, 5.79 eq.) was added. The reaction mixture was stirred at 25 degrees celsius for 0.5 hours. After the reaction is completed, the reaction solution is directly concentrated to obtain the hydrochloride of the compound 7-3.

Step C: the compound is prepared1-18a (10 mg, 37.99. Mu. Mol, 1 eq.) was dissolved in tetrahydrofuran (1 ml), and N, N-carbonyldiimidazole (7.39 mg, 45.59. Mu. Mol, 1 eq.) and triethylamine (11.53 mg, 113.79. Mu. Mol, 15.86. Mu.l, 3 eq.) were added. The reaction solution was stirred at 25 degrees celsius for 3 hours. Carbonyl diimidazole (6.16 mg, 37.99 μmol, 1 eq.) was added and the reaction mixture stirred at 25 degrees celsius for 2 hours. Compound 7-3 (17.15 mg 75.98 μmol, 2 eq, hydrochloride) was added and the reaction mixture stirred at 50 degrees celsius for 24 hours. After completion of the reaction, the reaction mixture was diluted with water (5 ml) and extracted with ethyl acetate (5 ml×2). The combined organic phases were washed with saturated brine (5 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the crude product which was subjected to high performance liquid chromatography (column; phenomenex Synergi C; 150 x 25 mm x 10 μm; mobile phase [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile: %28% -58%,10 min) to give compound 7.MS (ESI) m/z:479.2[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.53-12.49(m,1H),8.79(s,2H),7.82(s,1H),6.52-6.49(m,1H),5.86–5.73(m,2H),4.57-4.54(m,3H),3.85-3.83(m,4H),3.47(br s,4H),1.35(d,J＝6.8Hz,3H)

Compound 7 was tested by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 7 was 1.689min and the e.e. value was 99.104%.

Example 8

Step A: compound 8-1 (1.17 g, 8.86 mmol, 1.1 eq) was dissolved in isopropanol (10 ml) and compound 1-9 (1.5 g, 8.05 mmol, 1 eq) and N, N-diisopropylethylamine (2.08 g, 16.11 mmol, 2.81 ml, 2 eq) were added. The reaction mixture was stirred at 120 degrees celsius for 0.5 hours. After the reaction was completed, the reaction mixture was cooled to 25℃and water (50 mlLiter), stirring for 0.5 hour at 25 ℃, filtering the reaction solution, and spin-drying the filter cake in vacuum to obtain the compound 8-2.MS (ESI) m/z:183.0[ M+H ] ⁺ -100]. ¹ H NMR (400 MHz, chloroform-d) delta ppm 8.22 (s, 2H) 3.80-3.73 (m, 4H) 3.54-3.48 (m, 4H) 1.51 (s, 9H).

And (B) step (B): compound 8-2 (0.5 g, 1.77 mmol, 1 eq.) was dissolved in ethyl acetate (5 ml) and ethyl acetate hydrochloride (4 mol, 5 ml, 11.29 eq.) was added. The reaction mixture was stirred at 25 degrees celsius for 0.5 hours. After the reaction is completed, the reaction solution is filtered, and the filter cake is dried in vacuum to obtain the hydrochloride of the compound 8-3. MS (ESI) m/z:183.0[ M+H ] ⁺ ].

Step C: compound 1-18a (20 mg, 75.98 micromolar, 1 eq) was dissolved in tetrahydrofuran (1 ml), N-carbonyldiimidazole (12.32 mg, 75.98 micromolar, 1 eq) and triethylamine (23.07 mg, 227.95 micromolar, 31.73 micromolar, 3 eq) were added, and the reaction mixture was stirred at 25 degrees celsius for 1 hour, and compound 8-3 (16.61 mg, 75.98 micromolar, 1 eq, hydrochloride) was added to the reaction solution. The reaction mixture was stirred at 25 degrees celsius for 12 hours. After completion of the reaction, water (10 ml) was added to the reaction mixture, followed by extraction with ethyl acetate (10 ml. Times.3). The combined organic phases were washed with saturated brine (10 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated, and the crude product was purified by high performance liquid chromatography (column: phenomenex Synergi C, 150 x 25 mm x 10 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 34% -61%,9 min) to give compound 8.MS (ESI) m/z:472.1[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.55(s,1H),8.47(s,2H),7.82(s,1H),6.56-6.49(m,1H),5.88-5.70(m,2H),4.60-4.50(m,3H),3.71-3.64(m,4H),3.49-3.41(m,4H),1.32(d,J＝8.0Hz,3H).

Compound 8 was detected by SFC [ column number: chiralpak AD-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 8 was 2.069min and the e.e. value was 100%.

Example 9

Step A: compound 9-1 (100 mg, 701.54 μmol, 1 eq) and compound 1-9 (130.66 mg, 701.54 mmol, 1 eq) were dissolved in tetrahydrofuran (5 ml) and N, N-diisopropylethylamine (181.34 mg, 1.40 mmol, 244.39 μmol, 2 eq) was added. The reaction mixture was stirred at 25 degrees celsius for 1 hour. After completion of the reaction, water (50 ml) was added to the reaction mixture, followed by extraction with ethyl acetate (50 ml. Times.2). The combined organic phases were washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give compound 9-2.MS (ESI) m/z:193.1[ M+H ] ⁺ -100].

And (B) step (B): compound 9-2 (100 mg, 342.08 micromol, 1 eq) was dissolved in dichloromethane (5 ml) and diethylaminosulfur trifluoride (110.28 mg, 684.15 micromol, 90.39 μl, 2 eq) was added. The reaction mixture was stirred at 25 degrees celsius for 13 hours. After completion of the reaction, water (30 ml) was added to the reaction mixture, followed by extraction with ethyl acetate (30 ml. Times.2). The combined organic phases were washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated in vacuo to give crude product which was purified by thin layer chromatography (silica gel, petroleum ether/ethyl acetate=4/1) to give compound 9-3.MS (ESI) m/z:259.0[ M+H ] ⁺ -56].

Step C Compound 9-3 (60 mg, 190.88. Mu. Mol, 1 eq.) was dissolved in dioxane hydrochloride (4 mol, 5 ml). The reaction mixture was stirred at 25 degrees celsius for 15 minutes. After the reaction is completed, the reaction solution is directly concentrated to obtain the hydrochloride of the compound 9-4. MS (ESI) m/z:215.0[ M+H ] ⁺ ].

Step D: compounds 1-18a (15 mg, 56.99. Mu. Mol, 1 eq) were dissolved in N, N-dimethylformamide (1 ml), and N, N-carbonyldiimidazole (18.48 mg, 113.97. Mu. Mol, 2 eq) and triethylamine (28.83 mg, 284.98. Mu. Mol, 39.66. Mu.l, 5 eq) were added. The reaction solution was taken at 25Stirred at reflux for 2 hours. Compound 9-4 (28.57 mg, 113.97 μmol, 2 eq, hydrochloride) was added and the reaction mixture stirred at 25 degrees celsius for 24 hours. After completion of the reaction, water (30 ml) was added to the reaction mixture, followed by extraction with ethyl acetate (30 ml. Times.2). The combined organic phases were washed with saturated brine (30 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the crude product which was subjected to high performance liquid chromatography (column; unisil 3-100 C18 Ultra 150*50 mm. Times.3 μm; mobile phase [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 28% -58%,10 min) to give compound 9.MS (ESI) m/z:504.3[ M+H ] ⁺ ]. ¹ H NMR (400 MHz, chloroform-d) delta ppm 9.99 (brs, 1H), 8.45 (s, 2H), 7.58 (s, 1H), 6.59 (t, J=56.0 Hz, 1H) 5.84-5.73 (m, 2H), 5.55-5.40 (m, 1H), 4.71-4.61 (m, 2H), 4.35-4.22 (m, 1H), 3.93-3.90 (m, 4H), 3.61-3.57 (m, 4H), 1.46 (d, J=6.8 Hz, 3H).

Compound 9 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 9 was 1.313 min and the e.e. value was 100%.

Example 10

Step A: compound 10-1 (100 mg, 430.99 μmol, 1 eq) was dissolved in N-methylpyrrolidone (2 ml) and cesium carbonate (280.85 mg, 861.99 mmol, 2 eq) compound 1-9 (80.27 mg, 430.99 mmol, 1 eq) was added. The reaction mixture was stirred at 110 degrees celsius for 1 hour. After the completion of the reaction, the reaction mixture was cooled to 25℃and was then washed with ethyl acetate (10 ml), water (10 ml. Times.3) and saturated brine (10 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give Compound 10-2.MS (ESI) m/z:282.0[ M+H ] ⁺ -56]

And (B) step (B): compound 10-2 (190 mg, 536.20 micromolar, 1 eq.) was dissolved in ethyl acetate (2 Ml) ethyl acetate hydrochloride (4 mol/l, 2 ml) was added. The reaction mixture was stirred at 25 degrees celsius for 0.5 hours. After the reaction is completed, the reaction solution is directly concentrated to obtain the hydrochloride of the compound 10-3. MS (ESI) m/z:238.0[ M+H ] ⁺ ]

Step C: compounds 1-18a (15 mg, 56.99 micromolar, 1 eq) were dissolved in tetrahydrofuran (2 ml) and N, N-carbonyldiimidazole (18.48 mg, 113.97 micromolar, 2 eq) and triethylamine (23.07 mg, 227.95 micromolar, 31.73 μl, 4 eq) were added. The reaction was stirred at 50 degrees celsius for 4 hours. Carbonyl diimidazole (13.86 mg, 85.48. Mu. Mol, 1.5 eq.) was added and the reaction mixture stirred at 50℃for 12 hours, carbonyl diimidazole (13.86 mg, 85.48. Mu. Mol, 1.5 eq.) was added and the reaction mixture stirred at 25℃for 3 hours. Compound 10-3 (18.72 mg 68.38 micromolar, 1.2 eq, hydrochloride) was then added and the reaction mixture stirred at 50 degrees celsius for 24 hours. After the reaction was completed, the reaction solution was cooled to 25 degrees celsius, diluted with water (5 ml), and extracted with ethyl acetate (5 ml×3). The combined organic phases were washed with saturated brine (5 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the crude product which was subjected to high performance liquid chromatography (column; phenomenex Synergi C; 150×25 mm; 10 μm; mobile phase [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 40% -70%,10 min) to give compound 10.MS (ESI) m/z:527.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.55(brs,1H),7.82-7.75(m,2H),6.52-6.50(m,1H),5.86–5.74(m,2H),4.55-4.54(m,3H),3.52(s,8H),1.31(d,J＝6.8Hz,3H).

Compound 10 was tested by SFC [ column model: chiralcel OD-3X 4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 10 was 1.540min and the e.e. value was 100%.

Example 11

Step A: to a solution of compounds 1-9 (0.5 g, 3.89 mmol, 1 eq.) in N-methylpyrrolidone (10 ml) was added potassium carbonate (1.08 g, 7.78 mmol, 2 eq.) and compound 11-1 (724.38 mg, 3.89 mmol, 1 eq.) and the mixture was stirred at 100 degrees celsius for 12 hours. After the reaction solution was cooled to room temperature, water (50 ml) was added thereto, and a white solid was precipitated, filtered and the cake was collected, and dried under reduced pressure to obtain compound 11-2.MS (ESI) m/z 223.2[ M+H ] ⁺ -56].

And (B) step (B): to a solution of 11-2 (1 g, 3.59 mmol, 1 eq.) in ethyl acetate (10 ml) was added ethyl acetate hydrochloride (4 mol per liter, 10 ml, 11.13 eq.) and the mixture was stirred at 25 degrees celsius for 0.5 hours. The reaction solution was directly dried under reduced pressure to obtain hydrochloride of Compound 11-3. MS (ESI) m/z 179.2[ M+H ] ⁺ ].

Step C: to a solution of compounds 1-13 (12.94 g, 57.73 mmol, 11.45 ml, 1 eq.) in tetrahydrofuran (30 ml) was added lithium chloride (2.94 g, 69.28 mmol, 1.42 ml, 1.2 eq.) and triethylamine (7.01 g, 69.28 mmol, 9.64 ml, 1.2 eq.) and the mixture was stirred at 0 ℃ for 0.5 hours, then a solution of compounds 1-12 (10 g, 57.73 mmol, 1 eq.) in tetrahydrofuran (50 ml) was added and the mixture was stirred at 25 ℃ for 6 hours. The reaction solution was added to water (300 ml), extracted with ethyl acetate (300 ml. Times.2), and the organic phases were combined, washed with saturated brine (300 ml. Times.2), dried over anhydrous sodium sulfate, and concentrated to give compound 11-4.

Step D: to a solution of compound 11-4 (14 g, 57.54 mmol, 1 eq.) in ethyl acetate (80 ml) was added ethyl acetate hydrochloride (4 mol per liter, 80 ml, 5.56 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour. The reaction solution was concentrated directly to obtain hydrochloride of Compound 11-5.

Step E: to a solution of compounds 1-7 (36.72 g, 115.23 mmol, 1 eq.) in N, N-dimethylformamide (250 ml) was added triethylamine (34.98 g, 345.68 mmol, 48.11 ml, 3 eq.) and compounds 11-5 (20.7 g, 115.23 mmol, 1 eq., hydrochloride) and the mixture was stirred at 25 degrees celsius for 12 hours. The reaction mixture was diluted with ethyl acetate (500 ml), washed with water (500 ml. Times.3), then with saturated brine (500 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated, and the crude product was purified by high performance liquid chromatography (column: phenomenex luna C, 250, 80 mm, 10 μm; mobile phase: [ water (formic acid) -acetonitrile ]: acetonitrile%: 50% -80%,20 min) to give compound 11-6.MS (ESI) m/z 426.2[ M+H ] ⁺ ].

Step F: to a solution of compound 11-6 (10 g, 23.51 mmol, 1 eq.) in trifluoroacetic acid (100 ml) was added trifluoromethanesulfonic acid (17.00 g, 113.28 mmol, 10 ml, 4.82 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour. To the reaction solution was added saturated aqueous sodium carbonate solution to adjust the pH to 5, extracted with ethyl acetate (300 ml. Times.5), the organic phases were combined, washed with saturated brine (500 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by column chromatography (silica gel column, eluent gradient 0 to 50% petroleum ether/ethyl acetate, 100 ml/min) to give compound 11-7.MS (ESI) m/z 305.9[ M+H ] ⁺ ].

Step G: to a solution of compounds 11-7 (5 g, 16.38 mmol, 1 eq.) in tetrahydrofuran (50 ml) at 0 degrees celsius was added diisobutylaluminum hydride (1 mol per liter, 49.14 ml, 3 eq.) and the mixture was stirred at 10 degrees celsius for 1 hour. To the reaction solution was added a saturated solution of potassium sodium tartrate (50 ml), stirred at 25℃for 1 hour, extracted with ethyl acetate (100 ml. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 11-8.

Compounds 11-8 were tested by SFC [ column type number: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is isopropanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 11-8 was 1.146min and the e.e. value was 95.652%.

Step H: to a solution of compound 11-8 (3 g, 11.40 mmol, 1 eq.) in toluene (30 ml) was added 1, 8-diazacycloundec-7-ene (2.08 g, 13.68 mmol, 2.06 ml, 1.2 eq.) and diphenyl azide phosphate (3.76 g, 13.68 mmol, 2.96 ml, 1.2 eq.) and the mixture was stirred at 80 degrees celsius for 1 hour. The reaction solution was diluted with water (100 ml), extracted with ethyl acetate (100 ml×2), and the organic phases were combined, washed with saturated brine (200 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by column chromatography (silica, petroleum ether/ethyl acetate=3/1, 1/1) to give compounds 11-9.

Step I: to a solution of compounds 11-9 (2 g, 6.94 mmol, 1 eq.) in tetrahydrofuran (20 ml) was added triphenylphosphine (2.73 g, 10.41 mmol, 1.5 eq.) and the mixture was stirred at 40 degrees celsius for 0.5 hours. Water (3 ml) was further added to the reaction mixture, and the mixture was stirred at 40℃for 1.5 hours. Water (50 ml) was added for dilution, extraction with ethyl acetate (50 ml×2), the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated, and the crude product was purified by column chromatography (silica, petroleum ether/ethyl acetate=1/1, ethyl acetate/methanol=5/1) to give compounds 11-10.MS (ESI) m/z 263.15[ M+H ] ⁺ ]Step H: to a solution of compound 11-10 (50 mg, 190.67. Mu. Mol, 1 eq.) in N, N-dimethylformamide (1 ml) were added N, N-carbonyldiimidazole (37.10 mg, 228.81. Mu. Mol, 1.2 eq.) and triethylamine (77.18 mg, 762.69. Mu. Mol, 106.16. Mu. Mol, 4 eq.) and the mixture was stirred at 25℃for 1 hour, then compound 11-3 (40.94 mg, 190.67. Mu. Mol, 989.50. Mu. Mol, 1 eq., hydrochloride) was added and the mixture was stirred at 25℃for 1 hour. Diluting with water (30 ml), extracting with ethyl acetate (30 ml. Times.2), mixing the organic phases, washing with saturated saline (30 ml. Times.2), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to high performance liquid chromatography (column: phenomenex Synergi C, 150X 25 ml)Meter 10 microns; mobile phase [ water (formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 23% -41%,9 min) to give compound 11.MS (ESI) m/z 467.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.50(s,1H),8.23(s,2H),7.82(s,1H),6.77-6.46(m,1H),6.44(br dd,J＝3.2Hz,1H),5.65-5.60(m,2H),4.49(br d,J＝5.6Hz,1H),3.67-3.50(m,6H),3.36(br s,4H),2.08(s,3H),1.29(d,J＝6.4Hz,3H).

Compound 11 was detected by SFC [ column model: OD-3 50X 4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 11 was 1.641min and the e.e. value was 96.060%.

Example 12

Step A: to a solution of compound 12-1 (200 mg, 942.12 μmol, 1 eq.) in N-methylpyrrolidone (2 ml) was added potassium carbonate (260.41 mg, 1.88 mmol, 2 eq.) and compound 1-8 (171.97 mg, 942.12 μmol, 1 eq.) and the mixture was stirred at 80 degrees celsius for 2 hours. After the reaction solution was cooled to room temperature, water (10 ml) was added thereto, and solids were precipitated, filtered and the cake was collected, and dried under reduced pressure to give compound 12-2.MS (ESI) m/z 303.1[ M+H ] ⁺ -56].

And (B) step (B): to a solution of 12-2 (310 mg, 865.06. Mu. Mol, 1 eq.) in ethyl acetate (3 ml) was added ethyl acetate hydrochloride (4 mol per liter, 3 ml, 13.87 eq.) and the mixture was stirred at 25℃for 1 hour. The reaction solution was directly dried under reduced pressure to obtain hydrochloride of Compound 12-3. MS (ESI) m/z 259.0[ M+H ] ⁺ -56].

Step C:to a solution of compound 11-10 (80 mg, 305.07 micromol, 1 eq.) in dichloromethane (2 ml) was added N-methylmorpholine (46.29 mg, 457.61 micromol, 50.31 μl, 1.5 eq.) and compound 12-4 (73.79 mg, 366.08 micromol, 1.2 eq.) and the mixture was stirred at 25 degrees celsius for 12 hours. Water (20 ml) was added to dilute, and dichloromethane (20 ml. Times.2) was used to extract, and the organic phases were combined, washed with saturated brine (20 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 12-5.MS (ESI) m/z 428.1[ M+H ] ⁺ ].

Step D: to a solution of compound 12-5 (110 mg, 257.41. Mu. Mol, 1 eq.) in N, N-dimethylformamide (3 ml) were added compound 12-3 (91.03 mg, 308.89. Mu. Mol, 1.2 eq. Hydrochloride) and triethylamine (31.26 mg, 308.89. Mu. Mol, 42.99. Mu. L, 1.2 eq.) and the mixture was stirred at 25℃for 12 hours. Ethyl acetate (20 ml) was added to dilute, the organic phases were combined, washed with water (20 ml x 2), washed with saturated brine (20 ml x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated, the crude product was purified by high performance liquid chromatography (column: phenomenex Synergi C, 150 x 25 mm x 10 μm; mobile phase: [ water (formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 37% -67%,10 min) and then chiral resolved (column: DAICEL CHIRALCEL OD (250 mm x 30 mm, 10 μm); mobile phase: [0.1% Ammonia isopropyl alcohol ]]The method comprises the steps of carrying out a first treatment on the surface of the Isopropyl alcohol%: 30% -30%,3.3 minutes) to give compound 12.MS (ESI) m/z 547.3[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.48(br s,1H),8.70(s,2H),7.82(s,1H),6.93–6.90(m,1H),6.45-6.44(m,1H),5.64-5.60(m,2H),4.51-4.35(m,1H),4.37(br d,J＝10.4Hz,4H),3.69-3.66(t,2H),3.10(br d,J＝12.4Hz,2H),1.77-1.75(m,2H),1.52-1.50(m,2H),1.25(d,J＝6.8Hz,3H).

Compound 12 was detected by SFC [ column model: chiralcel OD-3X 4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is isopropanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 12 was 1.675min and the e.e. value was 97.136%.

Example 13

Step A: to a solution of compound 1-9 (1 g, 5.48 mmol, 1 eq.) and compound 13-1 (1.02 g, 5.48 mmol, 1 eq.) in N-methylpyrrolidone (20 ml) was added potassium carbonate (1.51 g, 10.96 mmol, 2 eq.) and the mixture was stirred at 100 degrees celsius for 12 hours. After the reaction solution was cooled to room temperature, water (100 ml) was added thereto, and the mixture was stirred at 25℃for 10 minutes, and a white solid was precipitated, filtered and a cake was collected and dried under reduced pressure to obtain Compound 13-2.MS (ESI) m/z 277.1[ M+H ] ⁺ -56].

And (B) step (B): to a solution of 13-2 (1.6 g, 4.81 mmol, 1 eq.) in ethyl acetate (10 ml) was added ethyl acetate hydrochloride (4 mol per liter, 10 ml, 8.31 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour. The reaction solution was directly dried under reduced pressure to obtain hydrochloride of Compound 13-3. MS (ESI) m/z 233.1[ M+H ] ⁺ ].

Step C: to a solution of compound 11-10 (50 mg, 190.67. Mu. Mol, 1 eq.) in N, N-dimethylformamide (3 ml) were added N, N-carbonyldiimidazole (37.10 mg, 228.81. Mu. Mol, 1.2 eq.) and triethylamine (77.18 mg, 762.69. Mu. Mol, 106.16. Mu. L, 4 eq.) and the mixture was stirred at 25℃for 1 hour, then compound 13-3 (51.23 mg, 190.67. Mu. Mol, 1 eq., hydrochloride) was added and the mixture was stirred at 40℃for 3 hours. Diluting with water (30 ml), extracting with ethyl acetate (30 ml. Times.2), mixing the organic phases, washing with saturated saline (30 ml. Times.2), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to high performance liquid chromatography (column: phenomenex Synergi C, 150X 25 mm. Times.10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the 25% -55% acetonitrile, 10 min) to give compound 13.MS (ESI) m/z 521.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.50(s,1H),7.83-7.81(m,2H),7.42(d,J＝9.6Hz,1H),6.82–6.80(m,1H),6.44–6.43(br dd,J＝3.6Hz,1H),5.65-5.60(m,2H),4.51-4.50(m,1H),3.72-3.66(m,4H),3.46–3.45(m,2H),3.45-3.40(m,4H),1.29(d,J＝6.4Hz,3H).

Compound 13 was detected by SFC [ column number: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 13 was 1.475min and the e.e. value was 96.192%.

Example 14

Step A: to a solution of compound 11-8 (2.15 g, 8.17 mmol, 1 eq.) in dimethyl sulfoxide (20 ml) was added 2-iodoxybenzoic acid (4.57 g, 16.34 mmol, 2 eq.) and the reaction stirred at 25 degrees celsius for 24 hours. The reaction solution was poured into water (80 ml), extracted with ethyl acetate (80 ml×2), and the organic phases were combined, washed with saturated brine (80 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1 to 1/1) to give compound 14-1.MS (ESI) m/z 262.1[ M+H ] ⁺ ].

And (B) step (B): to a solution of compound 14-1 (230 mg, 880.55. Mu. Mol, 1 eq.) in 1, 2-dichloroethane (6 ml) and tetrahydrofuran (6 ml) was added methylamine hydrochloride (594.53 mg, 8.81 mmol, 10 eq.) and the reaction was stirred at 25℃for 12 hours, then sodium borohydride acetate (466.56 mg, 2.20 mmol, 2.5 eq.) was added and the reaction was stirred at 25℃for a further 12 hours. The reaction solution was concentrated and the crude product was purified by reverse preparation (column: 330g Flash Column Welch Ultimate XB_C18 20-40 μm;120A, sample dissolution solvent: approximately 2.00 g sample dissolved in 10 ml methanol, flow rate: 100 ml/min, mobile phase: acetonitrile/water, gradient water%: 0-30%30 min; 30-100%25 min; instrument: TELEDYNE ISCO CombiFlashRf) to give compound 14-2.MS (ESI) m/z 277.1[ M+H ] ⁺ ]。

Step C: to a solution of compound 14-2 (80 mg, 289.58. Mu. Mol, 1 eq.) in dichloromethane (2 ml) were added N-methylmorpholine (58.58 mg, 579.17. Mu. Mol, 63.68. Mu. Mol,2 eq.) and compound 12-4 (87.55 mg, 434.38. Mu. Mol, 1.5 eq.) and the reaction was stirred at 25℃for 12 hours. The reaction solution was poured into water (80 ml), extracted with ethyl acetate (80 ml×2), the organic phases were combined, washed with saturated brine (80 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by thin layer chromatography (petroleum ether/ethyl acetate=1/1) to give compound 14-3.MS (ESI) m/z 442.1[ M+H ] ⁺ ].

Step D: to a solution of compound 14-3 (40 mg, 90.63 μmol, 1 eq.) in N, N-dimethylformamide (1 ml) were added N, N-diisopropylethylamine (35.14 mg, 271.89 μmol, 47.36 μmol, 3 eq.) and compound 1-11 (48.70 mg, 181.26umol,2 eq., hydrochloride) and the reaction stirred at 100 degrees celsius for 12 hours. The reaction mixture was poured into water (30 ml), extracted with ethyl acetate (30 ml. Times.2), the organic phases were combined, washed with saturated brine (30 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by high performance liquid chromatography (column: phenomenex Synergi C, 150 x 25 mm. Times.10 μm; mobile phase: [ water (formic acid) -acetonitrile ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 40% -70%,10 min) and then chiral separation (column: DAICEL CHIRALCEL OJ (250 mm. Times.30 mm, 10 μm); mobile phase: [0.1% Ammonia Water methanol ]]The method comprises the steps of carrying out a first treatment on the surface of the Methanol%: 15% -15%,4.5 min), t=1.882 min, compound 14 is obtained. MS (ESI) m/z 535.3[ M+H ] ⁺ ]. ¹ H NMR (400 MHz, chloroform-d) delta ppm 10.82 (s, 1H), 8.51 (s, 2H), 7.62 (s, 1H), 5.82-5.61 (m, 2H), 5.52-5.41 (m, 1H), 4.35-4.22 (m, 1H), 3.96-3.90 (m, 4H), 3.89-3.75 (m, 2H), 3.28-3.40 (m, 4H), 2.82 (s, 3H), 1.45 (d, J=6.4 Hz, 3H).

Compound 14 was detected by SFC [ column model: chiralpak AD-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 14 was 1.878min and the e.e. value was 99.036%.

Example 15

Step A: to a solution of compound 1-8 (0.5 g, 2.74 mmol, 1 eq.) and compound 15-1 (548.61 mg, 2.74 mmol, 1 eq.) in N, N-dimethylformamide (8 ml) was added potassium carbonate (757.16 mg, 5.48 mmol, 2 eq.) and the reaction was heated to 80 degrees celsius and stirred for 2 hours. The reaction solution was poured into water (50 ml), extracted with ethyl acetate (50 ml×2), the organic phases were combined, washed with saturated brine (50 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 15-2.MS (ESI) m/z 247.1[ M+H ] ⁺ -100].

And (B) step (B): to a solution of compound 15-2 (1 g, 2.89 mmol, 1 eq.) in methanol (10 ml) was added ethyl acetate hydrochloride (10 ml, 4 mol/l) and the reaction stirred at 25 degrees celsius for 30 minutes. The reaction solution was concentrated to obtain hydrochloride of Compound 15-3. MS (ESI) m/z 247.1[ M+H ] ⁺ ].

Step C: to a solution of compounds 11-10 (60 mg, 228.81. Mu. Mol, 1 eq.) in N, N-dimethylformamide (1.5 ml) were added carbonyldiimidazole (55.65 mg, 343.22. Mu. Mol, 1.5 eq.) and N, N-diisopropylethylamine (147.86 mg, 1.14. Mu. Mol, 199.27. Mu.l, 5 eq.) and the mixture was stirred at 25℃for 1 hour, then 15-3 (129.37 mg, 457.62. Mu. Mol, 2 eq., hydrochloride) was added to the reaction solution, and the reaction solution was heated to 60℃and stirred for 12 hours. After cooling, the reaction mixture was directly subjected to high performance liquid chromatography (column: unisil 3-100 C18 Ultra 150*50 mm. 3 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 34% -64%,7 min) and then purified via chiral column (column: DAICEL CHIRALPAK AD (250 mm x 30 mm, 10 microns); mobile phase: [0.1% Ammonia isopropyl alcohol ]]The method comprises the steps of carrying out a first treatment on the surface of the Isopropyl alcohol%: 40% -40%,4.9 min) to give compound 15.MS (ESI) m/z 535.2[ M+H ] ⁺ ]. ¹ H NMR(400MHz,CHLOROFORM-d)δppm 10.28(br s,1H),8.50(s,2H),7.59(s,1H),5.80-5.69(m,1H),5.68-5.59(m,1H),5.50-5.39(m,1H),4.72-4.56(m,3H),4.30-4.18(m,2H),4.02–3.78(m,3H),3.40-3.31(m,1H),3.28-3.14(m,2H),1.43(d,J＝6.4Hz,3H),1.17(d,J＝6.8Hz,3H).

Compound 15 was detected by SFC [ column model: chiralpak AD-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is ethanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 15 was 3.164min and 3.507min and the e.e. value was 3.696%.

Example 16

Step A: to a solution of compound 16-1 (500 mg, 2.20 mmol, 1 eq.) and compound 1-9 (410.28 mg, 2.20 mmol, 1 eq.) in 1, 4-dioxane (5 ml) was added tris (dibenzylideneacetone) dipalladium (100.86 mg, 110.14 μmol, 0.05 eq.), 2-dicyclohexylphosphine-2, 6-diisopropyloxy-, 1-biphenyl (51.40 mg, 110.14 μmol, 0.05 eq.) and cesium carbonate (1.44 g, 4.41 mmol, 2 eq.) and the reaction was replaced with nitrogen, and then the reaction mixture was heated to 80 degrees celsius and stirred for 12 hours. The reaction solution was poured into water (60 ml), extracted with ethyl acetate (60 ml×2), the organic phases were combined, washed with saturated brine (60 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1 to 3/1) to give compound 16-2.MS (ESI) m/z 333.2[ M+H ] ⁺ ].

And (B) step (B): to the reaction flask were added compound 16-2 (520 mg, 1.56 mmol, 1 eq.) and ethyl acetate hydrochloride (20 ml, 4 mol/l) and the reaction was stirred at 25 degrees celsius for 15 minutes. The reaction solution was concentrated to obtain hydrochloride of Compound 16-3. MS (ESI) m/z 233.1[ M+H ] ⁺ ].

Step C: to compound 11-10 (60 mg, 228.81 micromolar, 1 eq.) of N, N-dimethylformamideTo a solution of (1 ml) were added N, N-carbonyldiimidazole (55.65 mg, 343.22. Mu. Mol, 1.5 eq.) and triethylamine (115.76 mg, 1.14 mmol, 159.23. Mu.l, 5 eq.) and the mixture was stirred at 25 ℃ for 1 hour, then 16-3 (129.37 mg, 457.62. Mu. Mol, 2 eq., hydrochloride) was added to the reaction solution, and the reaction solution was heated to 60 ℃ and stirred for 12 hours. The reaction solution was poured into water (50 ml), extracted with ethyl acetate (50 ml. Times.2), the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was purified by high performance liquid chromatography (column: YMC Triart C18 150 x 25 mm. Times.5 μm; mobile phase: [ water (formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 23% -53%,10 min) to give compound 16.MS (ESI) m/z 521.3[ M+H ] ⁺ ]. ¹ H NMR (400 MHz, chloroform-d) delta ppm10.20 (br s, 1H), 8.42 (s, 2H), 7.59 (s, 1H), 5.80-5.59 (m, 2H), 5.49-5.39 (m, 1H), 4.72-4.56 (m, 1H), 4.18-4.33 (m, 1H), 4.03-3.87 (m, 2H), 3.68-3.58 (m, 4H), 3.46-3.37 (m, 4H), 1.43 (d, J=6.8 Hz, 3H).

Compound 16 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 16 was 1.578min and the e.e. value was 96.492%.

Example 17

Step A: to a solution of compound 1-8 (1 g, 5.48 mmol, 1 eq.) and compound 17-1 (1.10 g, 5.48 mmol, 1.08 ml, 1 eq.) in N, N-dimethylformamide (15 ml) was added potassium carbonate (1.51 g, 10.96 mmol, 2 eq.) and the reaction was heated to 80 degrees celsius and stirred for 2 hours. The reaction solution was poured into water (80 ml), extracted with ethyl acetate (80 ml×2), the organic phases were combined, washed with saturated brine (80 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 17-2.MS (ESI) m/z 291.1[ M+H ] ⁺ -56].

And (B) step (B): to the reaction flask were added compound 17-2 (1.9 g, 5.49 mmol, 1 eq) and ethyl acetate hydrochloride (20 ml, 4 mol/l) and the reaction was stirred at 25 degrees celsius for 15 minutes. The reaction solution was concentrated to obtain hydrochloride of compound 17-3. MS (ESI) m/z 247.1[ M+H ] ⁺ ].

Step C: to a solution of compounds 11-10 (60 mg, 228.81. Mu. Mol, 1 eq.) in N, N-dimethylformamide (2 ml) were added N, N-carbonyldiimidazole (44.52 mg, 274.57. Mu. Mol, 1.2 eq.) and triethylamine (115.76 mg, 1.14. Mu. Mol, 159.23. Mu. Mol, 5 eq.) and the mixture was stirred at 25℃for 1 hour, then 17-3 (97.02 mg, 343.21. Mu. Mol, 1.5 eq., hydrochloride) was added to the reaction solution, and the reaction solution was stirred at 25℃for 12 hours. The reaction solution was poured into water (50 ml), extracted with ethyl acetate (50 ml. Times.2), the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was purified by high performance liquid chromatography (column: unisil 3-100 C18 Ultra 150*50 mm. Times.3 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 26% -56%,7 min) to give compound 17.MS (ESI) m/z 535.3[ M+H ] ⁺ ]. ¹ H NMR (400 MHz, chloroform-d) delta ppm11.69 (br s, 1H), 8.48 (s, 2H), 7.60 (s, 1H), 5.75-5.53 (m, 2H), 5.48-5.38 (m, 1H), 4.80 (br s, 1H), 4.29-4.16 (m, 1H), 3.99-3.92 (m, 2H), 3.90-3.79 (m, 4H), 3.60 (t, J=5.6 Hz, 2H), 3.39 (t, J=6.0 Hz, 2H), 2.02-1.93 (m, 2H), 1.39 (d, J=6.4 Hz, 3H).

Compound 17 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 17 was 1.041min and the e.e. value was 96.008%.

Example 18

Step A: to chemical combinationTo a solution of 1-9 (1.33 g, 7.17 mmol, 1 eq.) and 18-1 (1 g, 7.17 mmol, 1 eq.) in N-methylpyrrolidone (20 ml) was added potassium carbonate (1.49 g, 10.75 mmol, 1.5 eq.) and the reaction was heated to 80℃and stirred for 2 hours. The reaction solution was poured into water (100 ml), extracted with ethyl acetate (100 ml×2), the organic phases were combined, washed with saturated brine (100 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 18-2.MS (ESI) m/z 234.1[ M+H ] ⁺ -56].

And (B) step (B): to the reaction flask were added compound 18-2 (1.9 g, 6.57 mmol, 1 eq) and methanol hydrochloride (20 ml, 4 mol/l) and the reaction was stirred at 25 degrees celsius for 30 minutes. The reaction solution was concentrated to obtain hydrochloride of Compound 18-3. MS (ESI) m/z 190.2[ M+H ] ⁺ ]Step C: to a solution of compounds 11-10 (100 mg, 381.34. Mu. Mol, 1 eq.) in N, N-dimethylformamide (2 ml) were added N, N-carbonyldiimidazole (80.38 mg, 495.75. Mu. Mol, 1.3 eq.) and triethylamine (192.94 mg, 1.91. Mu. Mol, 265.39. Mu.l, 5 eq.) and the mixture was stirred at 25℃for 1 hour, then 18-3 (86.06 mg, 381.34. Mu. Mol, 1 eq., hydrochloride) was added to the reaction solution, and the reaction solution was stirred at 25℃for 12 hours. The reaction mixture was poured into water (40 ml), extracted with ethyl acetate (40 ml. Times.2), the organic phases were combined, washed with saturated brine (40 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was purified by high performance liquid chromatography (column: unisil 3-100 C18 Ultra 150*50 mm. Times.3 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 19% -49%,7 min) to give compound 18.MS (ESI) m/z 478.2[ M+H ] ⁺ ]。 ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.49(br s,1H),7.89(d,J＝10.0Hz,1H),7.82(s,1H),7.36(d,J＝9.6Hz,1H),6.82-6.79(m,1H),6.45–6.43(m,1H),5.71-5.55(m,2H),4.56-4.44(m,1H),3.78-3.70(m,4H),3.67-3.65(m,2H),3.49-3.41(m,4H),1.29(d,J＝6.4Hz,3H).

Compound 18 was detected by SFC [ column number: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 18 was 1.552min and the e.e. value was 100%.

Example 19

Step A: to a solution of compound 19-1 (3 g, 12.98 mmol, 1 eq.) and compound 1-9 (2.90 g, 15.58 mmol, 1.2 eq.) in 1, 4-dioxane (30 ml) was added 2-dicyclohexylphosphine-2, 6-diisopropyloxy-1, 1-biphenyl (605.79 mg, 1.30 mmol, 0.1 eq.), sodium tert-butoxide (2.50 g, 25.96 mmol, 2 eq.), tris (dibenzylideneacetone) dipalladium (594.39 mg, 649.10 μmol, 0.05 eq.) and, after three nitrogen substitutions, stirred at 100 ℃ for 12 hours, after completion of the reaction, 10 ml ethyl acetate was added, filtered, and the crude product obtained by concentrating the filtrate under reduced pressure was purified by silica gel thin layer chromatography (petroleum ether/ethyl acetate=1:1) to give compound 19-2. MS (ESI) m/z:337.2[ M+H ] ⁺ ].

And (B) step (B): to a solution of compound 19-2 (3 g, 8.92 mmol, 1 eq.) in tetrahydrofuran (10 ml) was added glacial acetic acid (10 ml) and water (10 ml) and the mixture was stirred at 70 degrees celsius for 12 hours after three nitrogen substitutions. After completion of the reaction, the pH of the reaction solution was adjusted to 7 with a saturated sodium carbonate solution, the reaction solution was extracted with ethyl acetate (30 ml. Times.3), washed with saturated brine (30 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (silica gel column, gradient of eluent 0 to 30% petroleum ether/ethyl acetate, 100 ml/min) to obtain compound 19-3.MS (ESI) m/z:253.2[ M+H ] ⁺ ].

Step C: to acetonitrile (10 ml) of compound 19-3 (771 mg, 3.06 mmol, 1 eq.) and compound 19-4 (815.90 mg, 3.06 mmol, 1 eq.) was added potassium fluoride (355.06 mg, 6.11 mmol, 143.17 μl, 2 eq.) and the mixture was stirred at 25 degrees celsius for 4 hours. After completion of the reaction, acetonitrile (5 ml) was added to the reaction solution, followed byThe residue obtained by vacuum drying the filtrate was purified by column chromatography (silica gel column, gradient of eluent 0 to 15% petroleum ether/ethyl acetate 100 ml/min) to give compound 19-5.MS (ESI) m/z 303.1[ M+H ] ⁺ ].

Step D: to a solution of compound 19-5 (885 mg, 2.93 mmol, 1 eq.) in ethyl acetate (6 ml) was added ethyl acetate hydrochloride (4 mol per liter, 6 ml, 8.2 eq.) and the mixture was stirred at 25 degrees celsius for 12 hours. After the reaction, the reaction solution was directly concentrated under reduced pressure to obtain hydrochloride of compound 19-6. MS (ESI) m/z:203.2[ M+H ] ⁺ ].

Step E: to a solution of compounds 11-10 (100 mg, 381.34. Mu. Mol, 1 eq.) in N, N-dimethylformamide (2 ml) were added triethylamine (192.94 mg, 1.91. Mu. Mol, 265.39. Mu. L, 5 eq.) and N, N-carbonyldiimidazole (80.38 mg, 495.75. Mu. Mol, 1.3 eq.) and, after stirring at 25℃for 1 hour, compounds 19-6 (109.22 mg, 457.61. Mu. Mol, 1.2 eq., hydrochloride) were added to the reaction solution, and the mixture was stirred at 25℃for 12 hours. After completion of the reaction, water (10 ml) was added to the reaction mixture, extracted with ethyl acetate (10 ml. Times.3), the organic phases were combined, washed with saturated brine (10 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a residue, which was purified by high performance liquid chromatography (column: phenomenex Synergi C; 150X 25 mm. 10 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 22% -42%,10 min) to give compound 19.MS (ESI) m/z 491.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.57-12.45(m,1H),8.00-7.93(m,1H),7.82(s,1H),7.70-7.35(m,1H),6.86-6.72(m,1H),6.54-6.39(m,1H),6.21-6.06(m,1H),5.74-5.52(m,2H),4.56-4.43(m,1H),3.65(br t,J＝4.8Hz,2H),3.45(br d,J＝4.4Hz,1H),3.40-3.35(m,4H),3.11(br d,J＝4.8Hz,2H),2.85-2.78(m,1H),1.29(br d,J＝6.4Hz,3H).

Example 20

Step A: to a solution of compound 20-1 (500 mg, 2.50 mmol, 1 eq.) and compound 1-8 (455.70 mg, 2.50 mmol, 1 eq.) in N, N-dimethylformamide (5 ml) was added potassium carbonate (690.07 mg, 4.99 mmol, 2 eq.) and the reaction was heated to 80 degrees celsius and stirred for 1 hour. Water (15 ml) was added to the reaction solution, the reaction solution was stirred for 15 minutes at 25℃and filtered, and the cake was concentrated under reduced pressure to give compound 20-2.MS (ESI) m/z 291.1[ M+H ] ⁺ -56].

And (B) step (B): to the reaction flask were added compound 20-2 (460 mg, 1.33 mmol, 1 eq) and ethyl acetate (4 ml), ethyl acetate hydrochloride (4 ml, 4 mol/l, 12.05 eq) and the reaction was stirred at 25 degrees celsius for 12 hours. The reaction solution was concentrated to obtain hydrochloride of compound 20-3. MS (ESI) m/z 247.1[ M+H ] ⁺ ].

Step C: to a solution of compounds 11-10 (100 mg, 381.34. Mu. Mol, 1 eq.) in N, N-dimethylformamide (5 ml) were added N, N-carbonyldiimidazole (80.38 mg, 495.75. Mu. Mol, 1.3 eq.) and triethylamine (192.94 mg, 1.91. Mu. Mol, 265.39. Mu.l, 5 eq.) and the mixture was stirred at 25℃for 1 hour, then 20-3 (129.36 mg, 457.61. Mu. Mol, 1.2 eq., hydrochloride) was added to the reaction solution, and the reaction solution was stirred at 25℃for 1 hour. The reaction mixture was poured into water (10 ml), extracted with ethyl acetate (10 ml. Times.3), the organic phases were combined, washed with saturated brine (10 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product obtained was subjected to chiral separation (column: DAICEL CHIRALCEL OD (250 mm. Times.30 mm, 10 μm); mobile phase: [0.1% aqueous ammonia methanol ]The method comprises the steps of carrying out a first treatment on the surface of the Methanol%: 20% -20%,4.1 min), t= 1.318min, to give compound 20.MS (ESI) m/z 535.2[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.49(s,1H),8.72(s,2H),7.81(s,1H),6.76(t,J＝5.6Hz,1H),6.42(br dd,J＝7.2,3.6Hz,1H),5.74-5.49(m,2H),4.80(td,J＝6.4,3.2Hz,1H),4.55-4.47(m,1H),4.45-4.38(m,1H),3.98(br d,J＝12.8Hz,1H),3.86(br d,J＝13.6Hz,1H),3.67(br t,J＝4.4Hz,2H),3.24-3.16(m,1H),3.07(dd,J＝13.6,4.0Hz,1H),2.92-2.82(m,1H),1.29(d,J＝6.4Hz,3H),1.10(d,J＝6.8Hz,3H).

Compound 20 was detected by SFC [ column model: OD-3 50X 4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 20 was 1.318min and the e.e. value was 100%.

Example 21

Step A: to a solution of compound 21-1 (500 mg, 2.50 mmol, 1 eq.) and compound 1-8 (455.70 mg, 2.50 mmol, 1 eq.) in N, N-dimethylformamide (5 ml) was added potassium carbonate (690.07 mg, 4.99 mmol, 2 eq.) and the reaction was heated to 80 degrees celsius and stirred for 1 hour. Water (15 ml) was added to the reaction solution, the reaction solution was stirred at 25℃for 15 minutes, the reaction solution was filtered, and the cake was concentrated under reduced pressure to give Compound 21-2.MS (ESI) m/z 291.1[ M+H ] ⁺ -56].

And (B) step (B): to the reaction flask were added compound 21-2 (437 mg, 1.26 mmol, 1 eq) and ethyl acetate (4 ml), ethyl acetate hydrochloride (4 ml, 4 mol per liter, 12.68 eq) and the reaction solution was stirred at 25 degrees celsius for 12 hours. The reaction solution was concentrated to obtain hydrochloride of compound 21-3. MS (ESI) m/z 247.2[ M+H ] ⁺ ].

Step C: to a solution of compounds 11-10 (100 mg, 381.34. Mu. Mol, 1 eq.) in N, N-dimethylformamide (5 ml) were added N, N-carbonyldiimidazole (80.38 mg, 495.75. Mu. Mol, 1.3 eq.) and triethylamine (192.94 mg, 1.91. Mu. Mol, 265.39. Mu.l, 5 eq.) and the mixture was stirred at 25℃for 1 hour, then 21-3 (129.36 mg, 457.61. Mu. Mol, 1.2 eq., hydrochloride) was added to the reaction solution, and the reaction solution was stirred at 25℃for 1 hour. The reaction solution was poured into water (10 ml), extracted with ethyl acetate (10 ml×3), and the organic phases were combined, followed by extraction with saturated brine (10 mlX 2) washing, drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the obtained crude product to chiral separation (column: DAICEL CHIRAL Pak IG (250 mm x 30 mm, 10 μm); mobile phase: [0.1% Ammonia Water methanol ]]The method comprises the steps of carrying out a first treatment on the surface of the Methanol%: 30% -30%,5.7 min), t= 2.511min, to give compound 21.MS (ESI) m/z 535.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.49(s,1H),8.72(s,2H),7.81(s,1H),6.76(t,J＝5.6Hz,1H),6.42(br dd,J＝7.6,3.6Hz,1H),5.71-5.46(m,2H),4.80(td,J＝6.0,3.2Hz,1H),4.55-4.46(m,1H),4.44-4.38(m,1H),3.98(br d,J＝13.2Hz,1H),3.86(br d,J＝13.2Hz,1H),3.72-3.62(m,2H),3.26-3.15(m,1H),3.07(dd,J＝13.6,3.6Hz,1H),2.87(dt,J＝12.4,3.6Hz,1H),1.29(d,J＝6.4Hz,3H),1.10(d,J＝6.4Hz,3H).

Compound 21 was detected by SFC [ column model: chiralpak IG-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 20% -20%) to obtain: the retention time of compound 21 was 2.511min and the e.e. value was 100%.

Example 22

Step A: to a solution of compound 22-1 (500 mg, 1.62 mmol, 1 eq.) and compound 1-8 (354.19 mg, 1.94 mmol, 1.2 eq.) in 1, 4-dioxane (5 ml) was added bis (triphenylphosphine) palladium (II) dichloride (113.50 mg, 161.70 μmol, 0.1 eq.), cesium carbonate (2.11 g, 6.47 mmol, 4 eq.) in water (1 ml), after three nitrogen substitutions, stirred at 100 ℃ for 12 hours, 10 ml ethyl acetate was added after the reaction was completed, filtered, and the filtrate was concentrated under reduced pressure to give a crude product which was purified by silica gel column chromatography (silica gel column, eluent gradient 0-8% petroleum ether/ethyl acetate, 100 ml/min) to give compound 22-2.MS (ESI) m/z 274.1[ M+H ] ⁺ -56].

And (B) step (B): to the reaction flask were added compound 22-2 (420 mg, 1.28 mmol, 1 eq.) and ethanol (4 ml), palladium on carbon (80 mg, purity: 10%), and the reaction mixture was stirred for 1 hour at 25℃under the protection of hydrogen (15 pounds per square inch) after three hydrogen substitutions. The reaction solution was filtered and the filtrate was concentrated to obtain compound 22-3.MS (ESI) m/z 276.1[ M+H+ -56].

Step C: to the reaction flask were added compound 22-3 (375 mg, 1.13 mmol, 1 eq.) and ethyl acetate (2 ml), ethyl acetate hydrochloride (2 ml, 4 mol/l) and the reaction was stirred at 25 degrees celsius for 12 hours. The reaction mixture was concentrated to give hydrochloride of compound 22-4. MS (ESI) m/z 232.2[ M+H ] ⁺ ].

Step D: to a solution of compounds 11-10 (100 mg, 381.34. Mu. Mol, 1 eq.) in N, N-dimethylformamide (3 ml) were added N, N-carbonyldiimidazole (80.38 mg, 495.75. Mu. Mol, 1.3 eq.) and triethylamine (192.94 mg, 1.91. Mu. Mol, 265.39. Mu.l, 5 eq.) and the mixture was stirred at 25℃for 1 hour, then 22-4 (122.49 mg, 457.61. Mu. Mol, 1.2 eq., hydrochloride) was added to the reaction solution, and the reaction solution was stirred at 25℃for 1 hour. The reaction mixture was poured into water (10 ml), extracted with ethyl acetate (10 ml. Times.3), the organic phases were combined, washed with saturated brine (10 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the residue was purified by high performance liquid chromatography (column: phenomenex Synergi C, 150 x 25 mm. Times.10 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 28% -58%,10 min) to give compound 22.MS (ESI) m/z 520.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO- d ₆ )δppm 12.48(br s,1H),9.21(s,2H),7.82(s,1H),6.70(br s,1H),6.42(br d,J＝2.4Hz,1H),5.80-5.42(m,2H),4.50(br d,J＝5.6Hz,1H),4.03(br d,J＝12.8Hz,2H),3.65(br s,2H),3.16-3.08(m,1H),2.83(br t,J＝12.4Hz,2H),1.92(br d,J＝12.0Hz,2H),1.72-1.53(m,2H),1.29(br d,J＝6.4Hz,3H).

Compound 22 was detected by SFC [ column model: chiralpak IG-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 22 was 2.322min and the e.e. value was 96.406%.

Example 23

Step A: to a solution of compound 23-1 (1 g, 5.32 mmol, 1 eq.) and compound 1-9 (1.19 g, 6.38 mmol, 1.2 eq.) in 1, 4-dioxane (10 ml) was added (R) - (+) -2, 2-bis (diphenylphosphino) -1, 1-binaphthyl (331.13 mg, 531.79. Mu. Mol, 0.1 eq.), cesium carbonate (2.60 g, 7.98 mmol, 1.5 eq.) palladium acetate (119.39 mg, 531.79. Mu. Mol, 0.1 eq.) and 1-9 (1.19 g, 6.38 mmol, 1 eq.) under nitrogen were replaced three times and stirred at 100 ℃ for 12 hours, after completion of the reaction 10 ml of ethyl acetate was added, filtered, and the filtrate was concentrated under reduced pressure to give a crude product which was subjected to high performance liquid chromatography (column: phenomenex luna C: 18 x 70 x 10 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 35% -65%,20 min) to give compound 23-2.MS (ESI) m/z 294.1[ M+H ] ⁺ ].

And (B) step (B): to the reaction flask were added compound 23-2 (300 mg, 1.02 mmol, 1 eq) and ethyl acetate (3 ml), ethyl acetate hydrochloride (3 ml, 4 mol per liter, 11.74 eq) and the reaction was stirred at 25 degrees celsius for 12 hours. The reaction solution was concentrated to obtain hydrochloride of compound 23-3. MS (ESI) m/z 194.1[ M+H ] ⁺ ].

Step C: to a solution of compound 11-10 (100 mg, 381.34. Mu. Mol, 1 eq) in N, N-dimethylformamide (3 ml) were added N, N-carbonyldiimidazole (80.38 mg, 495.75. Mu. Mol, 1.3 eq) and triethylamine (192.94 mg, 1.91. Mu. Mol, 265.39. Mu.l, 5 eq) and the mixture was stirred at 25℃for 0.5 hours to give a solution of compound 23-4 in N, N-dimethylformamide which was used directly in the next step. MS (ESI) m/z 357.1[ M+H ] ⁺ ].

Step D: compound 23-3 (105.12 mg, 457.60 μmol, 1.2 eq, hydrochloride) was added to a solution of compound 23-4 (135.87 mg, 381.33 μmol, 1 eq) in N, N-dimethylformamide and the reaction stirred at 25 degrees celsius for 12 hours. The reaction mixture was poured into water (10 ml), extracted with ethyl acetate (10 ml. Times.3), the organic phases were combined, washed with saturated brine (10 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the residue was purified by high performance liquid chromatography (column: phenomenex Synergi C, 150 x 25 mm. Times.10 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 31% -51%,10 min) to give compound 23.MS (ESI) m/z 482.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.50(br s,1H),7.81(s,1H),7.62(d,J＝4.4Hz,1H),6.85(br t,J＝5.2Hz,1H),6.44(br d,J＝3.6Hz,1H),6.30(d,J＝4.4Hz,1H),5.70-5.51(m,2H),4.57-4.42(m,1H),3.65(br t,J＝4.4Hz,2H),3.48-3.43(m,4H),3.21(br t,J＝4.8Hz,4H),1.29(d,J＝6.4Hz,3H).

Compound 23 was detected by SFC [ column number: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 23 was 1.524min and the e.e. value was 96.872%.

Example 24

Step A: to a solution of compounds 1-13 (6.11 g, 27.26 mmol, 2.5 eq.) in acetonitrile (45 ml) at 0 degrees celsius were added lithium chloride (1.16 g, 27.26 mmol, 2.5 eq.) and N, N-diisopropylethylamine (3.52 g, 27.26 mmol, 2.5 eq.) and the mixture was stirred at 0 degrees celsius for 0.5 hours. A solution of compound 24-1 (2.5 g, 10.90 mmol, 1.0 eq.) in acetonitrile (30 ml) was slowly dropped into the system at 0 degrees celsius and the mixing system was stirred at 0 degrees celsius for a further 2.5 hours. Concentrating the reaction solution to dryness, and subjecting the residue to column chromatography (silica gel, petroleum ether/ethyl acetate=10/1, 5/1) to give compound 24-2.MS (ESI) m/z 322.1[ M+Na ⁺ ]. ¹ H NMR (400 MHz, chloroform-d) delta ppm 6.93-6.73 (m, 1H), 6.04-5.75 (m, 1H), 4.63-4.36 (m, 1H), 4.24-4.06 (m, 3H), 3.80 (dd, J=9.2 Hz, 1H), 1.66-1.58 (m, 3H), 1.57-1.41 (m, 12H), 1.32-1.25 (m, 3H).

And (B) step (B): to a solution of compound 24-2 (1.6 g, 5.34 mmol, 1 eq.) in dichloromethane (10 ml) at 25 degrees celsius was added trifluoroacetic acid (12.19 g, 206.89 mmol, 7.91 ml, 20 eq.) and water (2.21 g, 122.93 mmol, 2.21 ml, 23 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of Compound 24-3. MS (ESI) m/z 159.1[ M+H ] ⁺ ].

Step C: to a solution of compound 24-3 (2 g, 5.16 mmol, 1 eq, trifluoroacetate salt) in N, N-dimethylformamide (50 ml) at 25 degrees celsius were added compound 1-7 (1.65 g, 5.16 mmol, 1 eq) and triethylamine (2.61 g, 25.82 mmol, 3.59 ml, 5.0 eq) and the mixture was stirred at 25 degrees celsius for 1 hour. To the reaction solution was added water (200 ml), extracted with ethyl acetate (100 ml×2), and the organic phases were combined, washed with saturated brine (50 ml×4), dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by column chromatography (silica gel, petroleum ether/ethyl acetate=2/1, 1/1) to give compound 24-4.MS (ESI) m/z 442.0[ M+H ] ⁺ ].

Step D: to a solution of compound 24-4 (0.3 g, 675.38. Mu. Mol, 1 eq.) in trifluoroacetic acid (3 ml) at 0℃was added trifluoromethanesulfonic acid (510 mg, 3.4 mmol, 300. Mu. L, 5.03 eq.) and the mixture was stirred at 0℃for 0.5 hours. The reaction solution was slowly added dropwise to a saturated aqueous sodium hydrogencarbonate (50 ml) solution, stirred for 5 minutes, extracted with ethyl acetate (50 ml. Times.2), and the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 24-5.MS (ESI) M/z 322.2, [ M+H ] ⁺ ].

Step E: at 25 degrees celsius, to compound 2To a solution of 4-5 (0.2 g, 622.56. Mu. Mol, 1 eq.) in methylene chloride (5 ml) was added 3, 4-dihydropyran (78.55 mg, 933.85. Mu. Mol, 1.5 eq.) and trifluoroacetic acid (7.10 mg, 62.26. Mu. Mol, 0.1 eq.) and the mixture was stirred at 25℃for 2 hours. To the reaction solution was added water (50 ml), and after stirring for 5 minutes, extracted with ethyl acetate (50 ml. Times.2), and the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 24-6.MS (ESI) m/z 406.1[ M+H ] ⁺ ].

Step F: to a solution of compound 24-6 (240 mg, 592.05. Mu. Mol, 1 eq.) in tetrahydrofuran (5 ml) at 0℃under nitrogen, lithium diisobutylaluminum hydride (1 mol per liter, 1.78 ml, 3 eq.) was slowly added dropwise, and the mixture was stirred at 0℃under nitrogen for 0.5 hours. To the reaction solution was slowly added a saturated aqueous ammonium chloride (50 ml), stirred for 5 minutes, extracted with ethyl acetate (50 ml. Times.2), and the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 24-7.MS (ESI) m/z 364.0[ M+H ] ⁺ ].

Step G: to a solution of compound 24-7 (210 mg, 577.98. Mu. Mol, 1 eq.) in dichloromethane (5 ml) were added N-methylmorpholine (64.31 mg, 635.78. Mu. Mol, 69.90. Mu. Mol, 1.1 eq.) and compound 12-4 (128.15 mg, 635.78. Mu. Mol, 1.1 eq.) and the reaction was stirred at 25℃for 16 hours. To the reaction solution was added water (50 ml), and after stirring for 5 minutes, extracted with ethyl acetate (50 ml. Times.2), and the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 24-8.MS (ESI) m/z 529.2[ M+H ] ⁺ ].

Step H: to a solution of compound 24-8 (0.3 g, 567.71 micromol, 1 eq.) in N, N-dimethylformamide (5 ml) were added compound 1-11 (160.15 mg, 596.10 micromol, 1.05 eq.) and N, N-diisopropylethylamine (110.06 mg, 851.57 micromol, 148.33 μl, 1.5 eq.) and the reaction stirred at 25 ℃ for 3 hours. Reverse directionWater (50 ml) was added to the reaction solution, stirred for 5 minutes, extracted with ethyl acetate (50 ml. Times.2), and the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 24-9.MS (ESI) m/z 622.3[ M+H ] ⁺ ].

Step I: to a solution of compound 24-9 (0.3 g, 482.68. Mu. Mol, 1 eq.) in dichloromethane (5 ml) was added trifluoroacetic acid (1.54 g, 13.51 mmol, 1 ml, 27.98 eq.) and the mixture was stirred at 25℃for 1 hour. The reaction was concentrated and the residue was chromatographed using a high performance liquid chromatography (column: waters Xbridge C18 150 x 50 mm x 10 μm; mobile phase: [ water (ammonium bicarbonate) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 26% -56%,10 min) to give compound 24.MS (ESI) m/z 538.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 10.57–10.16(m,1H),8.52(s,2H),7.55(s,1H),6.43-6.12(m,1H),6.02–5.68(m,2H),4.69(d,J＝4.8Hz,2H),4.28(br s,1H),3.93(br t,J＝4.8Hz,5H),3.85-3.79(m,1H),3.61-3.55(m,4H),2.34-2.02(m,1H).

Compound 24 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 24 was 1.505min and the e.e. value was 100%.

Example 25

Step A: to a solution of compound 25-1 (1 g, 5.48 mmol, 662.25 μl, 1 eq.) in N-methylpyrrolidone (10 ml) was added compound 1-9 (1.02 g, 5.48 mmol, 1 eq.) and the mixture was stirred at 80 degrees celsius for 1 hour. Cooled to room temperature, water (50 ml) was added to the reaction mixture, and after stirring for 5 minutes, the above-mentioned mixture was filtered, and the cake was dried under reduced pressure to obtain compound 25-2.MS (ESI) m/z 333.1[ M+H ] ⁺ ].

And (B) step (B): will be converted intoCompound 25-2 (1 g, 3.01 mmol, 1 eq.) was added to a solution of ethyl acetate hydrochloride (4 mol per liter, 10 ml, 13.29 eq.) and the mixture stirred at 25 degrees celsius for 1 hour. Filtering and collecting filter cakes to obtain the hydrochloride of the compound 25-3. MS (ESI) m/z 233.2[ M+H ] ⁺ ].

Step D: to a solution of compound 23-4 (67.00 mg, 188.04 μmol, 1 eq.) in N, N-dimethylformamide (5 ml) was added compound 25-3 (60.40 mg, 225.65 μmol, 1.2 eq., hydrochloride) and triethylamine (57.08 mg, 564.13 μmol, 78.52 μmol, 3 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour. The reaction mixture was concentrated and the resulting residue was chromatographed using a high performance liquid chromatography (column: waters Xbridge C18 150 x 50 mm x 10 μm; mobile phase: [ water (ammonium bicarbonate) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Percent: 27% -27%,10 min), and chiral resolution (DAICEL CHIRALCEL OD (250 mm x 30 mm, 10 microns); mobile phase: [0.1% Ammonia Water methanol ]]The method comprises the steps of carrying out a first treatment on the surface of the Methanol%: 20% -20%,2.6 min) to give compound 25.MS (ESI) m/z 521.2[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.51(s,1H),8.71(d,J＝4.8Hz,1H),7.83(s,1H),7.05(d,J＝4.8Hz,1H),6.80(br s,1H),6.46(br s,1H),5.70-5.54(m,2H),4.51(br d,J＝5.6Hz,1H),3.76-3.71(m,4H),3.66(br d,J＝4.2Hz,2H),3.41-3.38(m,4H),1.30(br d,J＝6.4Hz,3H).

Compound 25 was detected by SFC [ column model: OD-3 50X 4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 25 was 1.413min and the e.e. value was 100%.

Example 26

Step A: to a solution of compound 1-9 (3 g, 16.11 mmol, 1 eq.) and compound 26-1 (2.36 g, 16.11 mmol, 1 eq., hydrochloride) in N, N-dimethylformamide (30 ml) was added N, N-diisopropylethylamine (208 g, 16.11 mmol, 2.81 ml, 1 eq.) and the mixture was stirred at 40 degrees celsius for 12 hours. Water (50 ml) was added to dilute, extraction was performed with ethyl acetate (50 ml. Times.2), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 26-2.MS (ESI) m/z 229.1[ M+H ] ⁺ ].

And (B) step (B): to a solution of 26-2 (4.8 g, 21.03 mmol, 1 eq.) in isopropanol (40 ml) under nitrogen, sodium tert-butoxide (6.06 g, 63.08 mmol, 3 eq.) and compound 26-3 (4.37 g, 21.03 mmol, 2.94 ml, 1 eq.) were added and the mixture stirred at 100 degrees celsius for 12 hours. The reaction solution was cooled to 25 degrees celsius, diluted with water (100 ml), extracted with ethyl acetate (100 ml×2), the organic phases combined, washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated, and the crude product purified by column chromatography (silica gel column, eluent gradient 0-30% petroleum ether/ethyl acetate, 80 ml/min) to give compound 26-4.MS (ESI) m/z 301.1[ M+H ] ⁺ -100].

Step C: to a solution of compound 26-4 (230 mg, 574.54. Mu. Mol, 1 eq.) in ethyl acetate (2 ml) was added ethyl acetate hydrochloride (4 mol per liter, 2 ml, 13.92 eq.) and the mixture was stirred at 25℃for 0.5 h. The reaction solution was directly dried under reduced pressure to obtain hydrochloride of Compound 26-5. MS (ESI) m/z 301.3[ M+H ] ⁺ ].

Step D: to a solution of compounds 11-10 (50 mg, 190.67 μmol, 1 eq) in N, N-dimethylformamide (3 ml) were added N, N-carbonyldiimidazole (37.10 mg, 228.81 μmol, 1.2 eq) and triethylamine (77.18 mg, 762.69 μmol, 106.16 μmol, 4 eq) and the mixture was stirred at 25 ℃ for 1 hour, then compound 26-5 (64.19 mg, 190.67 μmol, 1 eq, hydrochloride) was added and the mixture was stirred at 25 ℃ for 12 hours. Diluting with water (30 ml), extracting with ethyl acetate (30 ml. Times.2), mixing the organic phases, washing with saturated saline (30 ml. Times.2), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, and purifying the crude product by high performance liquid chromatography (column): phenomenex Synergi C18, 150 x 25 mm x 10 microns; mobile phase: [ Water (formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 45% -75%,2 min) to give compound 26.MS (ESI) m/z 589.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.50(s,1H),7.82(s,1H),7.46(s,1H),6.83–6.81(t,1H),6.45(br dd,J＝3.2Hz,1H),5.68-5.57(m,2H),4.52-4.48(m,1H),3.78-3.75(m,4H),3.67–3.66(m,2H),3.48-3.41(m,4H),1.29(d,J＝6.4Hz,3H).

Compound 26 was detected by SFC [ column number: chiralcel OD-3X 4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 26 was 1.118min and the e.e. value was 100%.

Example 27

Step A: to a solution of compound 11-6 (1.7 g, 4.00 mmol, 1 eq.) in ethanol (20 ml) was added palladium on carbon (0.1 g, 4.00 mmol, 10% purity), replaced three times with hydrogen and the mixture was stirred at 25 degrees celsius for 3 hours under protection of hydrogen. The reaction solution was filtered, and the filtrate was collected and concentrated to give compound 27-1.MS (ESI) m/z 428.2[ M+H ] ⁺ ].

And (B) step (B): to a solution of compound 27-1 (1.68 g, 3.93 mmol, 1 eq.) in tetrahydrofuran (20 ml) was added diisobutylaluminum hydride (1 mol per liter, 11.79 ml, 3 eq.) and the mixture was stirred at 0 ℃ for 1 hour. To the reaction solution was added a saturated solution of potassium sodium tartrate (60 ml), stirred at 25 ℃ for 1 hour, extracted with ethyl acetate (60 ml×2), the organic phases were combined, washed with saturated brine (60 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by column chromatography (silica, petroleum ether/ethyl acetate=3/1, 1/1) to give compound 27-2.

Step C: to compound 27-2 (0.9 g, 2.34 mmol)To a solution of dimethyl sulfoxide (10 ml) in 1 eq.) was added 2-iodoxybenzoic acid (784.73 mg, 2.80 mmol, 1.2 eq.) and the mixture was stirred at 60 ℃ for 2 hours, water (50 ml) was added to the reaction solution to dilute it, extraction was performed with ethyl acetate (50 ml×2), the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 27-3.MS (ESI) m/z 384.0[ M+H ] ⁺ ].

Step D: to a solution of compounds 1 to 13 (467.84 mg, 2.09 mmol, 414.02 μl, 1 eq.) in tetrahydrofuran (10 ml) were added lithium chloride (106.16 mg, 2.50 mmol, 51.29 μl, 1.2 eq.) and triethylamine (253.39 mg, 2.50 mmol, 348.54 μl, 1.2 eq.) and the mixture was stirred at 0 ℃ for 0.5 hours, then compound 27 to 3 (800 mg, 2.09 mmol, 1 eq.) was added to the reaction solution at 0 ℃ and the mixture was reacted at 25 ℃ for 1.5 hours. Water (50 ml) was added for dilution, extraction with ethyl acetate (50 ml×2), the organic phases were combined, washed with saturated brine (50 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by column chromatography (silica, petroleum ether/ethyl acetate=5/1, 3/1) to give compound 27-4.MS (ESI) m/z 454.1[ M+H ] ⁺ ].

Step E: to a solution of compound 27-4 (675 mg, 1.49 mmol, 1 eq.) in trifluoroacetic acid (10 ml) was added trifluoromethanesulfonic acid (1.70 g, 11.33 mmol, 1 ml, 7.61 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour. To the reaction solution was added water (50 ml) for dilution, saturated aqueous sodium carbonate solution was added for pH adjustment to 4, extracted with ethyl acetate (50 ml×2), the organic phases were combined, washed with saturated brine (50 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product by high performance liquid chromatography: (column: phenomenex luna C18150. Times.40 mm. Times.15. Mu.m; mobile phase: [ Water (formic acid) -acetonitrile)]: acetonitrile%: 28% -58%,10 min) to give compound 27-5.MS (ESI) m/z 334.2[ M+H ] ⁺ ].

Step F: to the formazan of compound 27-5 (150 mg, 450.04. Mu. Mol, 1 eq)To a solution of alcohol (2 ml) was added lithium hydroxide monohydrate (37.77 mg, 900.08 micromolar, 2 eq) and water (2 ml) and the mixture was stirred at 25 degrees celsius for 12 hours. Hydrochloric acid (1 mol/l) was added to the reaction solution to adjust the pH to 6, water (30 ml) was added to dilute, extraction was performed with ethyl acetate (30 ml. Times.2), the organic phases were combined, washed with saturated brine (30 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 27-6.MS (ESI) m/z 305.9[ M+H ] ⁺ ].

Step G: to a solution of compound 27-6 (50 mg, 163.80 μmol, 1 eq.) in tetrahydrofuran (2 ml) was added O- (7-azabenzotriazol-1-yl) -N, N-tetramethylurea hexafluorophosphine salt (93.42 mg, 245.70 μmol, 1.5 eq.) and triethylamine (66.30 mg, 655.20 μmol, 91.19 μmol, 5 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour. Compounds 1 to 11 (44.01 mg, 163.80. Mu. Mol, 1 eq. Hydrochloride) were then added to the reaction mixture and the mixture was stirred at 25℃for 2 hours. Diluting with water (30 ml), extracting with ethyl acetate (30 ml. Times.2), combining organic phases, washing with saturated brine (30 ml), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to HPLC (column: phenomenex Synergi C Ultra 150X 25 mm. 10 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 36% -66%,10 min) and then chiral resolved (column: REGIS (S, S) WHELK-O1 (250 mm. Times.25 mm, 10 μm); mobile phase: [0.1% Ammonia isopropyl alcohol ]]The method comprises the steps of carrying out a first treatment on the surface of the Isopropyl alcohol%: 40% -40%,5.2 min) to give compound 27.MS (ESI) m/z 520.2[ M+H ] ⁺ ].

Compound 27 was detected by SFC [ column model: (S, S) Whelk-Ol OD-3X 4.6mm I.D.,3.5 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is isopropanol (0.05% diethylamine); gradient (B%): 40% -40%) to obtain: the retention time of compound 27 was 2.457min and the e.e. value was 96.016%.

Example 28

Step A: to a solution of compound 28-1 (200 mg, 927.74. Mu. Mol, 1 eq.) in N-methylpyrrolidone (2 ml) was added compound 1-8 (168.79 mg, 927.74. Mu. Mol, 1 eq.) and potassium carbonate (255.61 mg, 1.85 mmol, 2 eq.) and the mixture was stirred at 100℃for 2 hours. After the reaction solution was cooled to 25 degrees centigrade, water (20 ml) was added, and the mixture was stirred at 25 degrees centigrade for 10 minutes, and the solid was precipitated, filtered and the filter cake was collected, and dried under reduced pressure to obtain compound 28-2.MS (ESI) m/z 307.1[ M+H ] ⁺ -56].

And (B) step (B): to a solution of 28-2 (360 mg, 993.52. Mu. Mol, 1 eq.) in ethyl acetate (2 ml) was added ethyl acetate hydrochloride (4 mol per liter, 2 ml) and the mixture was stirred at 25℃for 1 hour. The reaction solution was directly dried under reduced pressure to obtain hydrochloride of the compound 28-3. MS (ESI) m/z 263.1[ M+H ] ⁺ ].

Step C: to a solution of compound 11-10 (100 mg, 381.34. Mu. Mol, 1 eq.) in N, N-dimethylformamide (3 ml) were added N, N-carbonyldiimidazole (74.20 mg, 457.61. Mu. Mol, 1.2 eq.) and triethylamine (154.35 mg, 1.53. Mu. Mol, 212.31. Mu. L, 4 eq.) and the mixture was stirred at 25℃for 1 hour, then compound 28-3 (136.68 mg, 457.61. Mu. Mol, 1.2 eq., hydrochloride) was added and the mixture was stirred at 25℃for 11 hours. Diluting with water (20 ml), extracting with ethyl acetate (20 ml. Times.2), mixing the organic phases, washing with saturated saline (20 ml. Times.2), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to high performance liquid chromatography (column: phenomenex luna C, 150X 25 mm. Times.10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 28% -58%,2 min) to give compound 28-4.MS (ESI) m/z 551.1[ M+H ] ⁺ ].

Step D: compound 28-4 was isolated by chiral separation (column: phenomenex-Cellulose-2 (250 mm. Times.25 mm, 10 μm); mobile phase: [ acetonitrile/methanol (0.1% ammonia water) ]]The method comprises the steps of carrying out a first treatment on the surface of the Methanol%: 40% -40%,4.5 min) to give compound 28A and compound 28B.28A MS (ESI) m/z:551.4[M+H ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.49(br s,1H),8.71(s,2H),7.82(s,1H),6.70(t,J＝5.6Hz,1H),6.42(br dd,J＝7.2,3.2Hz,1H),5.77-5.50(m,2H),4.83(t,J＝5.6Hz,1H),4.71(br d,J＝2.0Hz,1H),4.54-4.32(m,2H),4.10-3.84(m,2H),3.66(br t,J＝4.8Hz,2H),3.56-3.44(m,2H),3.24-3.14(m,1H),3.04(dd,J＝4.0,13.6Hz,1H),2.96-2.84(m,1H),1.29(d,J＝6.4Hz,3H).

28B:MS(ESI)m/z:551.4[M+H ⁺ ].

Detection by SFC [ column model: uniChiral OZ-5H 50X 4.6mm I.D.,5 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is methanol+acetonitrile (0.05% diethylamine); gradient (B%): 40% -40%) to obtain: the retention time of compound 28A was 0.53 min, the e.e. value was 100%, the retention time of compound 28B was 0.769min, and the e.e. value was 99.756%.

Example 29

Step A: to a solution of compound 29-1 (0.75 g, 3.47 mmol, 1 eq.) in N-methylpyrrolidone (10 ml) was added compound 1-8 (632.98 mg, 3.47 mmol, 1 eq.) and potassium carbonate (958.56 mg, 6.94 mmol, 2 eq.) and the mixture was stirred at 100 degrees celsius for 2 hours. After the reaction solution was cooled to 25 ℃, water (50 ml) was added, and solids were precipitated, filtered and the cake was collected and dried under reduced pressure to give compound 29-2.MS (ESI) m/z 307.1[ M+H ] ⁺ -56].

And (B) step (B): to a solution of 29-2 (540 mg, 1.49 mmol, 1 eq.) in ethyl acetate (2 ml) was added ethyl acetate hydrochloride (4 mol per liter, 2 ml) and the mixture was stirred at 25 degrees celsius for 0.5 hours. The reaction solution was directly dried under reduced pressure to obtain hydrochloride of compound 29-3. MS (ESI) m/z 263.2[ M+H ] ⁺ ].

Step C: to compound 11-10 (150 mg, 572.01 micromolarTo a solution of N, N-dimethylformamide (5 ml) in 1 eq was added N, N-carbonyldiimidazole (111.30 mg, 686.41. Mu. Mol, 1.2 eq) and triethylamine (231.53 mg, 2.29. Mu. Mol, 318.47. Mu.l, 4 eq) and the mixture was stirred at 25℃for 1 hour, then compound 29-3 (170.85 mg, 572.01. Mu. Mol, 1 eq, hydrochloride) was added and the mixture was stirred at 25℃for 12 hours. Diluting with water (30 ml), extracting with ethyl acetate (30 ml. Times.2), mixing the organic phases, washing with saturated saline (30 ml), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to high performance liquid chromatography (column: phenomenex Synergi C, 150 x 25 mm. 10 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 25% -55%,10 min) to give compound 29-4.MS (ESI) m/z 551.2[ M+H ] ⁺ ].

Step D: compound 29-4 was isolated by chiral resolution (column: DAICEL CHIRALPAK AS (250 mm. Times.30 mm, 10 μm); mobile phase: [0.1% aqueous ammonia isopropanol)]The method comprises the steps of carrying out a first treatment on the surface of the Isopropyl alcohol%: 30% -30%,5.3 min) to give compound 29A and compound 29B.29A MS (ESI) m/z 551.3[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.49(br s,1H),8.71(s,2H),7.81(s,1H),6.65(br t,J＝5.6Hz,1H),6.42(br dd,J＝7.2,3.2Hz,1H),5.78-5.51(m,2H),4.76(t,J＝5.2Hz,1H),4.64(br d,J＝12.8Hz,1H),4.56-4.43(m,1H),4.33(br d,J＝13.2Hz,1H),4.09(br s,1H),3.85(br d,J＝13.2Hz,1H),3.66(br s,2H),3.35(br s,3H),3.22-3.02(m,2H),1.29(d,J＝6.4Hz,3H).

29B:MS(ESI)m/z:551.3[M+H ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.49(br s,1H),8.71(s,2H),7.82(s,1H),6.65(t,J＝5.6Hz,1H),6.42(br dd,J＝7.2,3.2Hz,1H),5.73-5.53(m,2H),4.75(t,J＝5.2Hz,1H),4.64(br d,J＝12.0Hz,1H),4.55-4.44(m,1H),4.40-4.28(m,1H),4.15-4.03(m,1H),3.91-3.80(m,1H),3.66(br s,2H),3.40-3.33(m,3H),3.22-3.02(m,2H),1.29(d,J＝6.4Hz,3H).

Detection by SFC [ column model: chiralpak AS-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is isopropanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 29A was 1.611 min and the e.e. value was 100%. The retention time of compound 29B was 1.893min and the e.e. value was 98.914%.

Example 30

Step A: to a solution of compound 30-1 (2 g, 14.43 mmol, 1 eq.) and compound 1-9 (2.69 g, 14.43 mmol, 1 eq.) in N, N-dimethylformamide (30 ml) was added potassium carbonate (1.99 g, 14.43 mmol, 1 eq.) and the mixture was stirred at 80 degrees celsius for 12 hours. After the reaction solution was cooled to room temperature, water (100 ml) was added thereto, and the mixture was stirred at 25℃for 10 minutes, and a solid was precipitated, filtered and a cake was collected, and dried under reduced pressure to obtain Compound 30-2.MS (ESI) m/z 233.1[ M+H ] ⁺ -56].

And (B) step (B): to a solution of 30-2 (1 g, 3.47 mmol, 1 eq.) in ethyl acetate (10 ml) was added ethyl acetate hydrochloride (4 mol, 10 ml, 11.53 eq.) and the mixture was stirred at 25 degrees celsius for 0.5 hours. The reaction solution was directly dried under reduced pressure to obtain hydrochloride of Compound 30-3. MS (ESI) m/z 189.2[ M+H ] ⁺ ].

Step C: to a solution of compound 11-8 (100 mg, 379.92. Mu. Mol, 1 eq.) in tetrahydrofuran (3 ml) were added N, N-carbonyldiimidazole (73.92 mg, 455.90. Mu. Mol, 1.2 eq.) and triethylamine (153.77 mg, 1.52. Mu. Mol, 211.52. Mu. L, 4 eq.) and the mixture was stirred at 40℃for 1 hour, then compound 30-3 (85.36 mg, 379.92. Mu. Mol, 1 eq., hydrochloride) was added and the mixture was stirred at 60℃for 24 hours. Diluting with water (30 ml), extracting with ethyl acetate (30 ml. Times.2), mixing the organic phases, washing with saturated saline (30 ml), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to high performance liquid chromatography (column: phenomenex Synergi C, 150 x 25 mm. 10 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 27% -57%,10 min) pureAnd (3) performing chemical reaction to obtain a compound 30.MS (ESI) m/z 478.1[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δppm 12.52(br s,1H),8.50(d,J＝2.4Hz,1H),7.92-7.85(m,1H),7.82(s,1H),6.93(d,J＝9.2Hz,1H),6.51(br dd,J＝3.6,7.2Hz,1H),5.90-5.69(m,2H),4.55(br d,J＝4.8Hz,3H),3.75-3.65(m,4H),3.47(br s,4H),1.31(d,J＝6.4Hz,3H).

Compound 30 was detected by SFC [ column model: chiralpak IC-3.50X4.6mm I.D,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is methanol+acetonitrile (0.05% diethylamine); gradient (B%): 40% -40%) to obtain: the retention time of compound 30 was 1.965min and the e.e. value was 100%.

Example 31

Step A: to a solution of compounds 1-7 (5 g, 15.69 mmol, 1 eq.) in trifluoroacetic acid (30 ml) was added trifluoromethanesulfonic acid (5.10 g, 33.98 mmol, 3 ml, 2.17 eq.) and the mixture was stirred at 25 degrees celsius for 0.5 hours. Diluting with water (100 ml), adding sodium carbonate to adjust pH to 7, extracting with ethyl acetate (100 ml. Times.2), combining organic phases, washing with saturated brine (100 ml), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, purifying the crude product by column chromatography (silica gel column, eluent gradient 0-100% petroleum ether/ethyl acetate, 80 ml/min), to give compound 31-1.

Step A: to a solution of compound 31-2 (10 g, 48.72 mmol, 1 eq.) in dimethyl sulfoxide (100 ml) was added 2-iodoacyl benzoic acid (27.29 g, 97.44 mmol, 2 eq.) and the reaction stirred at 25 degrees celsius for 12 hours. Ethyl acetate (200 ml) and water (200 ml) were added to the reaction solution and stirred for 5 minutes, the above mixed solution was filtered, the filtrate was extracted with ethyl acetate (100 ml), the organic phase was washed with water (200 ml×2), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 31-3.

And (B) step (B): to a solution of compounds 1 to 13 (9.49 g, 42.32 mmol, 8.40 ml, 1 eq.) in tetrahydrofuran (100 ml) were added triethylamine (5.14 g, 50.78 mmol, 7.07 ml, 1.2 eq.) and lithium chloride (2.15 g, 50.78 mmol, 1.04 ml, 1.2 eq.) and the mixture was stirred at 25 degrees celsius for 30 minutes, and a solution of compounds 31 to 3 (8.6 g, 42.32 mmol, 1 eq.) in tetrahydrofuran (80 ml) was added to the reaction solution which was stirred at 25 degrees celsius for 12 hours. The reaction solution was poured into water (200 ml), ethyl acetate (200 ml×2) was extracted, the organic phases were combined, washed with water (200 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=1/1, 3/1) to give compound 31-5.MS (ESI) m/z 174.2[ M+H ] ⁺ ]。 ¹ H NMR (400 MHz, chloroform-d) delta ppm 6.91 (dd, j=15.6, 4.8hz, 1H), 5.97 (dd, j=15.6, 1.6hz, 1H), 5.03-4.98 (m, 1H), 4.46 (br s, 1H), 4.24-4.15 (m, 2H), 3.50 (br d, j=4.03 hz, 2H), 3.35 (s, 3H), 1.45 (s, 9H), 1.29 (t, j=7.2 hz, 3H).

Step C: compound 31-5 (4.6 g, 16.83 mmol, 1 eq.) is dissolved in ethyl acetate hydrochloride (40 ml, 4 mol/l) and stirred for 15 minutes at 25 degrees celsius. The reaction solution was concentrated to obtain hydrochloride of Compound 31-6.

Step D: to a solution of compound 31-6 (1 g, 4.77 mmol, 1 eq. Hydrochloride) and compound 31-1 (946.87 mg, 4.77 mmol, 1 eq.) in acetonitrile (20 ml) was added potassium acetate (1.17 g, 11.92 mmol, 2.5 eq.) and the reaction was stirred at 25 ℃ for 48 hours. The reaction mixture was poured into water (100 ml) and extracted with ethyl acetate (100 ml. Times.2), the organic phases were combined, washed with saturated brine (100 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated, and the crude product was prepared by HPLC (column: phenomenex luna C, 150X 40 mm. 15. Mu.m; mobile phase: [ water (trifluoroacetic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile% :18% -48%,10 min)) to give compound 31-7.MS (ESI) m/z 336.1[ M+H ] ⁺ ]。

Step E: a solution of compounds 31-7 (6.67 g, 19.89 mmol, 1 eq.) in tetrahydrofuran (50 ml) was replaced three times with nitrogen at 0℃and diisobutylaluminum hydride (1 mol, 79.58 ml, 4 eq.) was added dropwise to the mixed system, and the mixed system was stirred at 0℃for 2 hours. A saturated solution of potassium sodium tartrate (50 ml) was added to the reaction solution, stirred at 25℃for 12 hours, diluted with water (150 ml), extracted with ethyl acetate (200 ml. Times.2), the organic phases were combined, washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, the filtrate concentrated, the crude product purified by column chromatography (silica gel column, gradient of eluent 0-100%, petroleum ether/ethyl acetate, 100 ml/min), followed by chiral resolution (column: DAICEL CHIRALPAK AD (250 mm. Times.30 mm, 10 μm); mobile phase: [ (0.1% ammonia) isopropanol)]The method comprises the steps of carrying out a first treatment on the surface of the Isopropyl alcohol%: 30% -30%,4.1 min) to give compound 31-8.MS (ESI) m/z 294.1[ M+H ] ⁺ ].

Step F: to a solution of compound 31-8 (0.4 g, 1.36 mmol, 1 eq.) in tetrahydrofuran (10 ml) and a solution of N, N-dimethylformamide (5 ml) were added N, N-carbonyldiimidazole (265.42 mg, 1.64 mmol, 1.2 eq.) and triethylamine (690.14 mg, 6.82 mmol, 949.30 μl, 5 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour, then compound 7-3 (369.41 mg, 1.64 mmol, 1.2 eq., hydrochloride) was added and the mixture was stirred at 70 degrees celsius for 6 hours. Diluting with water (80 ml), extracting with ethyl acetate (80 ml. Times.2), mixing the organic phases, washing with saturated saline (80 ml), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to high performance liquid chromatography (column: phenomenex luna C, 250X 25 mm, 10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 26% -56%,10 min) to give compound 31.MS (ESI) m/z 509.4[ M+H ] ⁺ ]. ¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.56(s,1H),8.77(s,2H),7.83(s,1H),6.47(br dd,J＝4.0,7.6Hz,1H),5.99-5.64(m,2H),4.75-4.63(m,1H),4.56(d,J＝4.8Hz,2H),3.84(dd,J＝4.4,6.4Hz,4H),3.64-3.42(m,6H),3.29(s,3H).

Compound 31 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 31 was 1.800min and the e.e. value was 100.000%.

Example 32

Step A: to a solution of compound 1-9 (500 mg, 2.68 mmol, 1 eq.) and compound 32-1 (449.53 mg, 3.22 mmol, 1.2 eq.) in N, N-dimethylformamide (5 ml) was added potassium carbonate (742.06 mg, 5.37 mmol, 2 eq.) and the reaction was heated to 80 degrees celsius and stirred for 1 hour. Water (15 ml) was added to the reaction solution, the reaction solution was stirred at 25℃for 15 minutes, the reaction solution was filtered, and the cake was concentrated under reduced pressure to give Compound 32-2.MS (ESI) m/z 234.1[ M+H ] ⁺ -56]。

And (B) step (B): to the reaction flask were added compound 32-2 (400 mg, 1.38 mmol, 1 eq) and ethyl acetate (2 ml), ethyl acetate hydrochloride (2 ml, 4 mol per liter, 5.79 eq) and the reaction was stirred at 25 degrees celsius for 12 hours. The reaction solution was concentrated to obtain hydrochloride of Compound 32-3. MS (ESI) m/z 190.1[ M+H ] ⁺ ]。

Step C: to a solution of compound 11-8 (100 mg, 379.92. Mu. Mol, 1 eq) in tetrahydrofuran (3 ml) were added N, N-carbonyldiimidazole (80.08 mg, 493.89. Mu. Mol, 1.3 eq) and triethylamine (192.22 mg, 1.90. Mu. Mol, 264.40. Mu.l, 5 eq), and the mixture was stirred at 60℃for 1 hour, after completion of the detection reaction, no work-up was required, to give compound 32-4 which was used directly in the next step.

Step D: compound 32-3 (102.89 mg, 455.90 micromolar, 1.2 eq, hydrochloride) was added to compound 32-4 (135.74 mg, 379.92 micromolar, 1 eq, tetrahydrofuran solution) and the reaction was stirred at 60 degrees celsius for 12 hours. The reaction mixture was poured into water (10 ml), extracted with ethyl acetate (10 ml. Times.3), the organic phases were combined, washed with saturated brine (10 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the residue was purified by high performance liquid chromatography (column: phenomenex Synergi C, 150 x 25 mm. Times.10 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the 31% -61% acetonitrile, 10 min) to give compound 32.MS (ESI) m/z 479.2[ M+H ] ⁺ ]。

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.51(br s,1H),8.66(d,J＝4.8Hz,1H),7.82(s,1H),7.18(d,J＝4.8Hz,1H),6.51(br dd,J＝7.2,3.6Hz,1H),5.93-5.66(m,2H),4.56(br s,2H),3.81-3.71(m,4H),3.45(br d,J＝6.4Hz,4H),1.31(d,J＝6.4Hz,3H),1.05(t,J＝7.2Hz,1H).

Compound 32 was detected by SFC [ column model: chiralcel OJ-3.50 x 4.6mm I.D.,3 microns; mobile phase: phase A is supercritical carbon dioxide and phase B is methanol (0.05% DEA); gradient (B%): 5% -40%) to obtain: the retention time of compound 32 was 1.748min and the e.e. value was 95.128%.

Example 33

Step A: to a solution of compound 1-8 (546.83 mg, 3.00 mmol, 1.2 eq.) and compound 33-1 (500 mg, 2.50 mmol, 1 eq.) in N, N-dimethylformamide (5 ml) was added potassium carbonate (690.07 mg, 4.99 mmol, 2 eq.) and the reaction was heated to 80 degrees celsius and stirred for 1 hour. Water (15 ml) was added to the reaction solution, the reaction solution was stirred at 25℃for 15 minutes, the reaction solution was filtered, and the cake was concentrated under reduced pressure to give Compound 33-2.MS (ESI) m/z 291.1[ M+H ] ⁺ -56]。

And (B) step (B): to the reaction flask were added compound 33-2 (786 mg, 2.27 mmol, 1 eq) and ethyl acetate (4 ml), ethyl acetate hydrochloride (4 ml, 4 mol per liter, 7.05 eq) and the reaction was stirred at 25 degrees celsius for 12 hours. The reaction solution was concentrated to obtain hydrochloride of compound 33-3. MS (ESI) m/z 247.1[ M+H ] ⁺ ]。

Step C: to a solution of compound 11-8 (200 mg, 759.83. Mu. Mol, 1 eq) in tetrahydrofuran (5 ml) were added N, N-carbonyldiimidazole (160.17 mg, 987.78. Mu. Mol, 1.3 eq) and triethylamine (384.44 mg, 3.80. Mu. Mol, 528.80. Mu.l, 5 eq), and the mixture was stirred at 60℃for 1 hour, after completion of the detection reaction, no work-up was required, to give compound 32-4 which was used directly in the next step.

Step D: compound 33-3 (257.76 mg, 911.80 μmol, 1.2 eq, hydrochloride) was added to compound 32-4 (271.48 mg, 759.84 μmol, 1 eq, tetrahydrofuran solution) and the reaction was stirred at 60 degrees celsius for 12 hours. The reaction mixture was poured into water (10 ml), extracted with ethyl acetate (10 ml. Times.3), the organic phases were combined, washed with saturated brine (10 ml. Times.2), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the residue was purified by high performance liquid chromatography (column: phenomenex C18X 30 mm. Times.3 μm; mobile phase: [ water (0.225% formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the B%:38% -68%,7 min) to give compound 33.MS (ESI) m/z 536.2[ M+H ] ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.52(s,1H),8.68(s,2H),7.81(s,1H),7.26(br d,J＝8.0Hz,1H),6.48(br d,J＝3.6Hz,1H),5.97-5.61(m,2H),4.54(br d,J＝13.2Hz,3H),4.45(br d,J＝4.8Hz,2H),3.72-3.55(m,1H),3.18(br t,J＝11.6Hz,2H),1.82(br dd,J＝3.6,13.0Hz,2H),1.30(br d,J＝6.4Hz,5H).

Compound 33 was detected by SFC [ column model: chiralpak AD-3.50 x 4.6mm i.d.,3 microns; mobile phase: phase A is supercritical carbon dioxide and phase B is methanol (0.05% DEA); gradient (B%): 5% -40%) to obtain: the retention time of compound 33 was 1.892min, the e.e. value was 97.612%;

example 34

Step A: to a solution of compound 31-8 (2 g, 6.82 mmol, 1 eq.) in tetrahydrofuran (20 ml) and N, N-dimethylformamide (10 ml) were added N, N' -carbonyldiimidazole (1.33 g, 8.18 mmol, 1.2 eq.) and triethylamine (3.45 g, 34.10 mmol, 4.75 ml, 5 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour, then compound 8-3 (1.79 g, 8.18 mmol, 1.2 eq., hydrochloride) was added and the mixture was stirred at 60 degrees celsius for 3 hours. Diluting with water (100 ml), extracting with ethyl acetate (100 ml. Times.2), mixing the organic phases, washing with saturated saline (100 ml), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate, and subjecting the crude product to high performance liquid chromatography (column: phenomenex luna C, 250X 70 mm, 10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 30% -60%,20 min) to give compound 34.MS (ESI) m/z 502.2[ M+H ] ⁺ ]. ¹ H NMR(400MHz, DMSO-d ₆ ) δ=12.56 (s, 1H), 8.48 (s, 2H), 7.83 (s, 1H), 6.48 (brdd, j=4.0, 7.6hz, 1H), 5.91-5.68 (m, 2H), 4.76-4.64 (m, 1H), 4.56 (d, j=4.8 hz, 2H), 3.69 (dd, j=4.4, 6.4hz, 4H), 3.61-3.40 (m, 6H), 3.28 (s, 3H) compound 34 is SFC tested [ column type number: chiralpak IG-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is methanol+acetonitrile (0.05% diethylamine); gradient (B%): 40% -40%) to obtain: the retention time of compound 34 was 1.316min and the e.e. value was 100.000%.

Example 35

Step A: to a solution of compound 31-8 (300 mg, 1.02 mmol, 1 eq.) in dichloromethane (10 ml) was added triethylamine (155.28 mg, 1.53 mmol, 213.59 μl, 1.5 eq.) and compound 12-4 (247.45 mg, 1.23 mmol, 1.2 eq.) and the mixture was stirred at 25 degrees celsius for 12 hours. The reaction solution was directly spin-dried to obtain compound 35-1.MS (ESI) m/z 459.1[ M+H ] ⁺ ].

And (B) step (B): to a solution of compound 35-2 (500.00 mg, 2.50 mmol, 1 eq.) in N, N-dimethylformamide (10 ml) was added potassium carbonate (345.04 mg, 2.50 mmol, 1 eq.) and compound 7-1 (348.37 mg, 2.50 mmol, 1 eq.) and the mixture was stirred at 80 degrees celsius for 12 hours. The reaction solution was cooled to 25 degrees celsius, water (50 ml) was added, a yellow solid precipitated, filtered, the filter cake was collected, and dried under reduced pressure to give compound 35-3.MS (ESI) m/z 204.2[ M+H ] ⁺ -100].

Step C: to a solution of compound 35-3 (490 mg, 1.62 mmol, 1 eq.) in ethyl acetate (5 ml) was added ethyl acetate hydrochloride (4 mol, 5 ml, 12.38 eq.) and the reaction was stirred at 25 degrees celsius for 12 hours. The reaction solution was concentrated to obtain hydrochloride of compound 35-4. MS (ESI) m/z 204.2[ M+H ] ⁺ ].

Step D: to a solution of compound 35-1 (200 mg, 436.35 micromol, 1 eq) and compound 35-4 (156.89 mg, 654.53 micromol, 1.5 eq, hydrochloride) in N, N-dimethylformamide (5 ml) was added triethylamine (88.31 mg, 872.70 micromol, 121.47 μl, 2 eq) and the mixture was stirred at 25 degrees celsius for 12 hours. The reaction solution was diluted with water (30 ml), extracted with ethyl acetate (30 ml×2), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was purified by high performance liquid chromatography (columnPhenomenex luna C18, 150 x 25 mm x 10 microns; mobile phase: [ Water (formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 36% -66%,10 min) to give compound 35.MS (ESI) m/z 523.4[ M+H ] ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.55(s,1H),8.90-8.51(m,2H),7.82(s,1H),6.58-6.29(m,1H),6.00-5.63(m,2H),4.75-4.64(m,1H),4.63-4.43(m,4H),4.36-4.23(m,1H),3.93-3.77(m,1H),3.59-3.45(m,2H),3.29(s,4H),3.19-3.05(m,2H),1.01(d,J＝6.8Hz,3H).

Compound 35 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is ethanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 35 was 1.681min and the e.e. value was 100.000%.

Example 36

Step A: to a solution of compound 36-2 (600.00 mg, 3.00 mmol, 1 eq.) in N, N-dimethylformamide (10 ml) was added potassium carbonate (414.04 mg, 3.00 mmol, 1 eq.) and compound 7-1 (418.05 mg, 3.00 mmol, 1 eq.) and the mixture was stirred at 80 degrees celsius for 12 hours. The reaction solution was cooled to 25℃and water (80 ml) was added, the solid precipitated, filtered, the cake was collected and dried under reduced pressure to give compound 36-3.MS (ESI) m/z 204.2[ M+H ] ⁺ -100].

And (B) step (B): to a solution of compound 36-3 (740 mg, 2.44 mmol, 1 eq.) in ethyl acetate (7 ml) was added ethyl acetate hydrochloride (4 mol, 7 ml, 11.48 eq.) and the reaction stirred at 25 degrees celsius for 12 hours. The reaction solution was concentrated to obtain hydrochloride of Compound 36-4. MS (ESI) m/z 204.3[ M+H ] ⁺ ].

Step C: to compound 35-1 (200 mg, 436.35. Mu. Mol, 1 eq.) and compound 36-4 (156.89 mg, 654.53. Mu. Mol, 1.5 eq., hydrochloride)To a solution of N, N-dimethylformamide (5 ml) was added triethylamine (88.31 mg, 872.71. Mu. Mol, 121.47. Mu.l, 2 eq.) and the mixture was stirred at 25℃for 12 hours. The reaction solution was diluted with water (30 ml), extracted with ethyl acetate (30 ml. Times.2), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was purified by high performance liquid chromatography (column: phenomenex luna C, 150 x 25 mm. Times.10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 36% -66%,10 min) to give compound 36.MS (ESI) m/z 523.4[ M+H ] ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.55(s,1H),8.77(s,2H),7.91-7.68(m,1H),6.58-6.37(m,1H),6.17-5.68(m,2H),4.75-4.63(m,1H),4.62-4.42(m,4H),4.36-4.24(m,1H),3.96-3.80(m,1H),3.60-3.45(m,2H),3.29(s,4H),3.20-3.01(m,2H),1.02(d,J＝6.8Hz,3H).

Compound 36 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is ethanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 36 was 1.744min and the e.e. value was 100.000%.

Example 37

Step A: lithium chloride (2.74 g, 64.59 mmol, 1.32 ml, 2.5 eq.) was dissolved in acetonitrile (40 ml), to the solution was added compound 37-2 (5.02 g, 25.84 mmol, 1 eq.) and N, N-diisopropylethylamine (8.35 g, 64.59 mmol, 2.5 eq.) at 25 degrees celsius, after stirring for 0.5 hours, a solution of compound 37-1 (3 g, 25.84 mmol, 1 eq.) in acetonitrile (40 ml) was added to the solution, and stirring was continued at 25 degrees celsius for 2.5 hours. After the completion of the reaction, water (50 ml) was added to the reaction mixture, stirred for 5 minutes, extracted with ethyl acetate (50 ml. Times.2), and the organic phase was washed with saturated brine (50 ml. Times.2),dried over anhydrous sodium sulfate, filtered and dried by spin-drying. The resulting residue was purified by silica gel column (petroleum ether: ethyl acetate=2:1 to 1:1) to give compound 37-3.MS (ESI) m/z 125.3[ M+H ] ⁺ -32].

¹ H NMR (400 MHz, chloroform-d) δ=6.90-6.72 (m, 1H), 6.19-5.99 (m, 1H), 3.70 (s, 3H), 2.62-2.45 (m, 4H), 2.24 (s, 3H).

And (B) step (B): compound 37-3 (1.7 g, 10.88 mmol, 1 eq.) was dissolved in tetrahydrofuran (20 ml) and water (20 ml), lithium hydroxide monohydrate (593.75 mg, 14.15 mmol, 1.3 eq.) was added and the reaction stirred at room temperature for 1 hour. After the completion of the reaction, the reaction mixture was adjusted to pH 5 with hydrochloric acid (2 mol/l), and the aqueous phase was extracted with ethyl acetate (50 ml. Times.4). The organic phases were combined, washed with saturated brine (60 ml. Times.2), dried over anhydrous sodium sulfate, and filtered and dried to give compound 37-4.MS (ESI) m/z 141.0[ M-H ] ⁺ ].

Step C: to a solution of compound 37-4 (750 mg, 5.28 mmol, 1 eq.) in tetrahydrofuran (30 ml) was added compound 7-3 (1.19 g, 5.28 mmol, 1 eq., hydrochloride), O- (7-azabenzotriazol-1-YL) -N, N-tetramethylurea hexafluorophosphine salt (2.21 g, 5.8 mmol, 1.1 eq.) and N, N-diisopropylethylamine (1.7 g, 13.19 mmol, 2.3 ml, 2.5 eq.) and the mixture was stirred at 25 degrees celsius for 16 hours. After completion of the reaction, water (50 ml) was added to the reaction mixture, followed by stirring for 5 minutes, filtration, extraction of the filtrate with ethyl acetate (50 ml. Times.2), washing of the combined organic phases with saturated brine (50 ml. Times.2), drying over anhydrous sodium sulfate, filtration, and spin-drying. The residue obtained was purified by silica gel column (petroleum ether: ethyl acetate=1:1 to 0:1) to give compound 37-5.MS (ESI) m/z 314.2[ M+H ] ⁺ ].

Step D: to a solution of compound 37-5 (0.5 g, 1.58 mmol, 1 eq.) in tetrahydrofuran (10 ml) was added tetraethyltitanate (1.81 g, 7.92 mmol, 1.64 ml, 5 eq.) and compound 37-6 (958.36 mg, 7.92 mmol, 5 eq.) and the mixture was stirred under nitrogen at 70 ℃ for 48 hours. After the reaction is finished, adding water into the reaction solution50 ml), stirred for 5 minutes, filtered, the filtrate extracted with ethyl acetate (50 ml×2), the organic phases combined, washed with saturated brine (50 ml×2), dried over anhydrous sodium sulfate, filtered, and spun-dried. Compound 37-7 is obtained. MS (ESI) m/z 417.3[ M+H ] ⁺ ].

Step E: to a solution of compounds 37-7 (0.5 g, 1.2 mmol, 1 eq.) in dichloromethane (10 ml) at 0 degrees celsius was slowly added sodium cyanoborohydride (75.43 mg, 1.2 mmol, 1 eq.) and acetic acid (36.04 mg, 600.18 μmol, 34.33 μl, 0.5 eq.) and the mixture was stirred at 0 degrees celsius for 1 hour. After the completion of the reaction, water (50 ml) was added to the reaction mixture, stirred for 5 minutes, the aqueous phase was extracted with ethyl acetate (50 ml. Times.2), and the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and dried by spinning. Compound 37-8 is obtained. MS (ESI) m/z 419.2[ M+H ] ⁺ ].

Step F: to a solution of compounds 37-8 (30 mg, 71.68. Mu. Mol, 1 eq.) in ethyl acetate (3 ml) was added ethyl acetate hydrochloride solution (4 mol per liter, 1.5 ml, 83.71 eq.) and the mixture was stirred at 25℃for 1 hour. After the reaction, concentrating and spin-drying to obtain the hydrochloride of the compound 37-9. MS (ESI) m/z 298.1[ M+H ] ⁺ -16].

Step G: to a solution of compound 31-1 (12.81 mg, 64.55 μmol, 1 eq.) in acetonitrile (3 ml) was added compound 37-9 (25 mg, 64.55 μmol, 1 eq., 2 hydrochloride) and sodium bicarbonate (16.27 mg, 193.65 μmol, 7.53 μmol, 3 eq.) and the mixture was stirred at 25 degrees celsius for 16 hours. After completion of the reaction, water (50 ml) was added to the reaction mixture, stirred for 5 minutes, the aqueous phase was extracted with ethyl acetate (50 ml. Times.2), and the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and dried by spinning. The residue was purified by high performance liquid chromatography (column: phenomenex luna C, 150 x 25 mm x 10 μm; mobile phase: [ water (formic acid) -acetonitrile ];% acetonitrile: 24% -54%,10 min) to give compound 37.

MS(ESI)m/z:477.2[M+H ⁺ ].

1H NMR(400MHz,DMSO-d ₆ )δ＝12.50(br s,1H),8.79(s,2H),7.81(s,1H),6.39(br d,J＝3.2Hz,1H),5.74-5.62(m,1H),5.61-5.50(m,1H),4.46(br d,J＝6.0Hz,1H),3.85(br d,J＝4.4Hz,2H),3.83-3.77(m,2H),3.55(br s,4H),2.43(br s,2H),2.25(q,J＝6.8Hz,2H),1.28(d,J＝6.8Hz,3H).

Example 38

Step A: to a solution of compound 38-1 (0.25 g, 1.62 mmol, 1 eq.) in N, N-dimethylformamide (20 ml) was added potassium carbonate (447.00 mg, 3.23 mmol, 2 eq.) and compound 1-9 (301.19 mg, 1.62 mmol, 1 eq.) and the mixture was stirred at 100 degrees celsius for 24 hours. The reaction solution was cooled to 25 degrees celsius, poured into water (100 ml), white solid precipitated, filtered, the filter cake was collected, and concentrated under reduced pressure to give compound 38-2.MS (ESI) m/z 249.2[ M+H ] ⁺ -56].

And (B) step (B): to a solution of compound 38-2 (360 mg, 1.18 mmol, 1 eq.) in ethyl acetate (5 ml) was added ethyl acetate hydrochloride (4 mol, 5 ml, 16.91 eq.) and the reaction stirred at 25 degrees celsius for 2 hours. The reaction mixture was concentrated to give hydrochloride of compound 38-3. MS (ESI) m/z 205.3[ M+H ] ⁺ ].

Step C: to a solution of compound 31-8 (80 mg, 272.81. Mu. Mol, 1 eq.) in tetrahydrofuran (2 ml) and a solution of N, N-dimethylformamide (2 ml) were added N, N-carbonyldiimidazole (53.08 mg, 327.37. Mu. Mol, 1.2 eq.) and triethylamine (138.03 mg, 1.36. Mu. Mol, 189.86. Mu.l, 5 eq.) and the mixture was stirred at 25℃for 2 hours, then compound 38-3 (78.81 mg, 327.37. Mu. Mol, 1.2 eq., hydrochloride) was added and the mixture was stirred at 60℃for 12 hours. The reaction solution was diluted with water (30 ml), extracted with ethyl acetate (30 ml×2), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude productBy high performance liquid chromatography (column: phenomenex luna C, 150, 25 mm. 10 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 42% -72%,10 min) to give compound 38.MS (ESI) m/z 524.3[ M+H ] ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.68-12.47(m,1H),8.22-8.10(m,2H),7.83(s,1H),6.53-6.41(m,1H),5.96-5.68(m,2H),4.73-4.64(m,1H),4.55(d,J＝4.8Hz,2H),3.72-3.64(m,4H),3.62-3.40(m,6H),3.28(s,3H),1.80-1.71(m,1H),0.91-0.83(m,2H),0.66-0.59(m,2H).

Compound 38 was detected by SFC [ column number: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is isopropanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 38 was 1.92min and the e.e. value was 100.000%.

Example 39

Step A: to a solution of compound 8-1 (0.5 g, 3.77 mmol, 1 eq.) in N, N-dimethylformamide (20 ml) was added potassium carbonate (1.04 g, 7.54 mmol, 1 eq.) and compound 35-2 (755.05 mg, 3.77 mmol, 1 eq.) and the mixture was stirred at 80 degrees celsius for 12 hours. The reaction solution was cooled to 25 degrees celsius, diluted with water (30 ml), extracted with ethyl acetate (30 ml×2), the organic phases combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated, and the crude product purified by column chromatography (silica gel column, eluent gradient 0-15%, petroleum ether/ethyl acetate, 100 ml/min) to give compound 39-1.MS (ESI) m/z 197.1[ M+H ] ⁺ -100].

And (B) step (B): to a solution of compound 39-1 (860 mg, 2.90 mmol, 1 eq.) in ethyl acetate (8 ml) was added ethyl acetate hydrochloride (4 mol, 8 ml, 11.03 eq.) and the reaction was stirred at 25 degrees celsius for 4 hours. Concentrating the reaction solution to obtain a salt of the compound 39-2 An acid salt. MS (ESI) m/z 197.2[ M+H ] ⁺ ].

Step C: to a solution of compound 35-1 (200 mg, 436.35 micromol, 1 eq) and compound 39-2 (152.30 mg, 654.53 micromol, 1.5 eq, hydrochloride) in N, N-dimethylformamide (5 ml) was added triethylamine (88.31 mg, 872.70 micromol, 121.47 μl, 2 eq) and the mixture was stirred at 25 degrees celsius for 12 hours. The reaction solution was diluted with water (30 ml), extracted with ethyl acetate (30 ml. Times.2), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was purified by high performance liquid chromatography (column: phenomenex luna C, 150 x 25 mm. Times.10 μm; mobile phase: [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 40% -70%,10 min) to give compound 39.MS (ESI) m/z 516.4[ M+H ] ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.54(br s,1H),8.45(s,2H),7.82(s,1H),6.54-6.38(m,1H),5.94-5.70(m,2H),4.74-4.64(m,1H),4.62-4.47(m,2H),4.46-4.31(m,2H),4.30-4.18(m,1H),3.85-3.80(m,1H),3.60-3.46(m,2H),3.28(s,3H),3.19-3.06(m,2H),2.99-2.92(m,1H),1.04(d,J＝6.4Hz,3H).

Compound 39 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is ethanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 39 was 1.420min and the e.e. value was 100.000%.

Example 40

Step A: to a solution of borane in tetrahydrofuran (1 mole per liter, 12.86 ml, 12.86 mmol, 3 eq) was added dropwise 40-1 (1 g, 4.29 mmol, 1 eq) in tetrahydrofuran (5 ml) at 0 degrees celsius and the mixture was stirred at 0 degrees celsius for 2 hours. After the detection reaction, methanol (10 ml) is slowly added into the reaction solution under stirring at 0 ℃ until no gas is discharged Ethyl acetate (100 ml) was added for dilution, and the mixture was washed with 1 mol/l hydrochloric acid (30 ml), followed by saturated brine (10 ml×2), dried over anhydrous sodium sulfate, filtered, and concentrated to give crude compound 40-2, which was used directly in the next reaction. ¹ H NMR (chloroform-d, 400 MHz): delta ppm 5.40-4.80 (m, 1H), 3.88-3.73 (m, 1H), 3.71-3.61 (m, 2H), 3.58-3.49 (m, 1H), 3.49-3.42 (m, 1H), 3.38 (s, 3H), 3.30-3.04 (m, 1H), 1.97-1.86 (m, 1H), 1.86-1.75 (m, 1H), 1.50-1.42 (m, 9H).

And (B) step (B): to a solution of 40-2 (0.68 g, 3.10 mmol, 1 eq.) in dichloromethane (20 ml) at 0 degrees celsius was added in portions dess-martin oxidant (1.32 g, 3.10 mmol, 1 eq.). The mixture was stirred at 25 degrees celsius for 12 hours. After the detection reaction is finished, the reaction solution is concentrated to be dry, and the residue is subjected to silica gel column chromatography (silica gel with 1000 meshes and eluent gradient ethyl acetate/petroleum ether 10/1 to 2/1) to obtain a compound 40-3. ¹ H NMR (chloroform-d, 400 MHz): delta (ppm) 9.56 (s, 1H), 5.66-5.32 (m, 1H), 4.19 (s, 1H), 3.55-3.38 (m, 2H), 3.28 (s, 3H), 2.20-1.95 (m, 2H), 1.46 (s, 9H).

Step C: to a solution of compounds 1-13 (846.15 mg, 3.77 mmol, 2 eq.) and diisopropylethylamine (731.69 mg, 5.66 mmol, 3 eq.) in acetonitrile (10 ml) at 0 degrees celsius was added lithium chloride (240.01 mg, 5.66 mmol, 3 eq.) and the mixture was stirred at 0 degrees celsius for half an hour. Subsequently, a solution of 40-3 (0.41 g, 1.89 mmol, 1 eq.) in acetonitrile (3 ml) was added dropwise to the above reaction solution. The reaction mixture was stirred at 25 degrees celsius for 12 hours. After completion of the reaction, 1 mol/l hydrochloric acid (50 ml) was added to the reaction mixture, which was extracted with ethyl acetate (80 ml. Times.2), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the obtained residue was purified by column chromatography (silica gel of 1000 mesh, eluent gradient ethyl acetate/petroleum ether 100/1 to 3/1) to give compound 40-4. ¹ H NMR (chloroform-d, 400 MHz): delta ppm 6.95-6.77 (m, 1H), 6.00-5.87 (m, 1H), 5.33-4.97 (m, 1H), 4.67-4.35 (m, 1H), 4.28-4.15 (m, 2H), 3.51-3.37 (m, 2H), 3.32 (s, 3H), 2.02-1.86 (m, 1H), 1.83-1.67 (m, 1H), 1.45 (s, 9H), 1.29 (s, 3H).

Step D: compound 40-4 (0.32 g, 1.11 mmol, 1 eq.) was stirred in a methanolic hydrogen chloride solution (4 mol per liter, 5 ml, 20 mmol, 17.96 eq.) at 0 degrees celsius for 2 hours. Concentrating to obtain crude compound 40-5.

Step E: compound 40-5 (0.208 g, 1.11 mmol, 1 eq.) compound 31-1 (0.15 g, 755.55 mmol, 0.68 eq.) and sodium bicarbonate (373.29 mg, 4.44 mmol, 4 eq.) in acetonitrile (8 ml) are stirred at 25 degrees celsius for 12 hours. After completion of the reaction, water (20 ml) was added to the reaction mixture, which was extracted with ethyl acetate (80 ml), washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the obtained residue was purified by thin layer chromatography (silica gel, eluent: ethyl acetate/petroleum ether=2/1) to give compound 40-6.MS (ESI) m/z 350.1[ M+H ] ⁺ ].

Step F: to a solution of compound 40-6 (0.155 g, 0.444 mmol, 1 eq.) in tetrahydrofuran (8 ml) was slowly added dropwise a solution of diisobutylaluminum hydride (1 mol per liter, 1.77 ml, 4 eq.) at 0 degrees celsius, and the mixed system was stirred at 0 degrees celsius for 3 hours. After completion of the detection reaction, a saturated aqueous solution (60 ml) of sodium potassium tartrate was added to the reaction mixture to quench it, stirring was continued for 12 hours, extraction was performed with ethyl acetate (100 ml. Times.2), the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the resulting residue was purified by column chromatography (silica 1000 mesh silica gel, eluent gradient ethyl acetate/petroleum ether 100/1 to 2/1) to give compound 40-7.MS (ESI) m/z 308.1[ M+H ] ⁺ ].

Step G: to a solution of compound 40-7 (36.00 mg, 117.16 μmol, 1 eq.) in tetrahydrofuran (5 ml) was added N, N' -carbonyldiimidazole (28.5 mg, 175.74 μmol, 1.5 eq.) and triethylamine (59.28 mg, 0.586 mmol, 5 eq.) and the mixture was stirred at 25 degrees celsius for 12 hours. Compound 7-3 (52.88 mg, 0.234 mmol, 2 eq, hydrochloride) was added to the reaction solution and the mixture was stirred at 25 degrees celsius for 12 hours. After completion of the detection reaction, water (40 ml) was added to the reaction mixture) Quench, extract with ethyl acetate (40 ml×2), combine the organic phases, wash with saturated brine (40 ml×2), dry over anhydrous sodium sulfate, filter, concentrate the filtrate and obtain a residue by high performance liquid chromatography (column: phenomenex luna C18, 150 x 25 mm x 10 microns; mobile phase: [ Water (formic acid) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 32% -62%,10 min) to give compound 40.MS (ESI) m/z 523.2[ M+H ] ⁺ ]. ¹ H NMR(CD ₃ OD,400 MHz): delta (ppm) 8.88-8.55 (m, 2H), 8.03-7.53 (m, 1H), 6.03-5.71 (m, 2H), 4.70-4.67 (m, 1H), 4.61 (br s, 2H), 3.95 (br s, 4H), 3.58 (br s, 6H), 3.35-3.45 (m, 3H), 2.25-2.04 (m, 1H), 1.80-2.01-1.80 (m, 1H). Compound 40 is tested by SFC [ column type number: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 40 was 1.741min and the e.e. value was 92.500%.

Example 41

Step A: to a solution of borane in tetrahydrofuran (1 mole per liter, 38.66 ml, 38.66 mmol, 3 eq.) was added dropwise a solution of 41-1 (2.8 g, 12.89 mmol, 1 eq.) in tetrahydrofuran (10 ml) at 0 degrees celsius and the mixture was stirred at 0-10 degrees celsius for 2 hours. After the completion of the detection reaction, methanol (30 ml) was slowly added to the reaction solution with stirring at 0℃until no gas was evolved, ethyl acetate (100 ml) was added for dilution, and the mixture was washed with 1 mol/l hydrochloric acid (50 ml), followed by washing with saturated sodium carbonate (50 ml) and brine (50 ml), drying over anhydrous sodium sulfate, and filtration and concentration to give a crude compound 41-2, which was directly used for the next reaction. ¹ H NMR(CD ₃ OD,400MHz):δ(ppm)3.58-3.51(m,1H),3.51-3.40(m,2H),1.66-1.49(m,2H),1.48-1.39(m,9H),1.39-1.28(m,2H),0.96(s,3H).

And (B) step (B): in a solution of 41-2 (2.1 g, 10.33 mmol, 1 eq.) in dichloromethane (20 ml) at 0deg.CThe dess-martin oxidizer (4.47 g, 10.54 mmol, 1.02 eq.) was added in portions. The mixture was stirred at 25 degrees celsius for 12 hours. After the detection reaction is finished, the reaction solution is concentrated to be dry, and the residue is subjected to silica gel column chromatography (silica gel with 1000 meshes and eluent gradient ethyl acetate/petroleum ether 10/1 to 2/1) to obtain the compound 41-3. ¹ H NMR (chloroform-d, 400 MHz) delta (ppm) 9.67-9.46 (m, 1H), 5.29 (br s, 1H), 4.35-4.15 (m, 1H), 1.58-1.53 (m, 2H), 1.46-1.43 (m, 9H), 1.43-1.32 (m, 2H), 0.96 (t, J=7.2 Hz, 3H).

Step C: to a solution of compounds 1-13 (3.41 g, 15.20 mmol, 2 eq.) and lithium chloride (966.75 mg, 22.81 mmol, 3 eq.) in acetonitrile (20 ml) at 0 degrees celsius was added N, N-diisopropylethylamine (2.95 g, 22.81 mmol, 3 eq.) and the mixture was stirred at 0 degrees celsius for half an hour. Subsequently, a solution of 41-3 (1.53 g, 7.60 mmol, 1 eq.) in acetonitrile (3 ml) was added dropwise to the above reaction solution. The reaction mixture was stirred at 25 degrees celsius for 12 hours. After completion of the reaction, 1 mol/l hydrochloric acid (50 ml) was added to the reaction mixture, which was extracted with ethyl acetate (80 ml. Times.2), the organic phases were combined, washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the obtained residue was purified by column chromatography (silica 1000 mesh silica gel, eluent gradient ethyl acetate/petroleum ether 100/1 to 3/1) to give compound 41-4. ¹ H NMR(CD ₃ OD,400 MHz): delta (ppm) 6.94-6.77 (m, 1H), 5.98-5.83 (m, 1H), 4.67-4.20 (m, 3H), 1.63-1.48 (m, 2H), 1.47 (s, 9H), 1.44 (s, 2H), 1.30 (t, j=7.2 hz, 3H), 0.96 (s, 3H). Step D: compound 41-4 (700 mg, 2.58 mmol, 1 eq.) was stirred in methanol solution of hydrogen chloride (4 mol per liter, 30 ml, 20 mmol, 46.52 eq.) at 25 degrees celsius for 2 hours. After the detection reaction is finished, concentrating to obtain a crude product of the compound 41-5.

Step E: compound 41-5 (0.44 g, 2.57 mmol, 1 eq.) compound 31-1 (0.46 g, 755.55 mmol, 0.9 eq.) and sodium bicarbonate (647.58 mg, 7.71 mmol, 3 eq.) in acetonitrile (10 ml) are stirred at 25 degrees celsius for 12 hours. After completion of the reaction, 1 mol/L hydrochloric acid (50 ml) was added to the reaction mixture, followed by ethyl acetate (80 ml. Times.2)Extraction, washing with saturated saline (30 ml. Times.2), drying over anhydrous sodium sulfate, filtering, concentrating the filtrate, and purifying the obtained residue by column chromatography (silica 1000 mesh silica gel, eluent gradient ethyl acetate/petroleum ether 100/1 to 2/1) to obtain compound 41-6.MS (ESI) m/z 334.1[ M+H ] ⁺ ].

Step F: to a solution of compound 41-6 (0.57 g, 1.71 mmol, 1 eq.) in tetrahydrofuran (15 ml) was slowly added dropwise a solution of diisobutylaluminum hydride (1 mol per liter, 6.84 ml, 4 eq.) at 0 degrees celsius and the mixture was stirred at 0 degrees celsius for 3 hours. After completion of the detection reaction, a saturated aqueous solution (60 ml) of sodium potassium tartrate was added to the reaction mixture to quench it, stirring was continued for 12 hours, extraction was performed with ethyl acetate (100 ml. Times.2), the organic phases were combined, washed with saturated brine (100 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the resulting residue was purified by column chromatography (silica 1000 mesh silica gel, eluent gradient ethyl acetate/petroleum ether 100/1 to 2/1) to give compound 41-7.MS (ESI) m/z 292.1[ M+H ] ⁺ ].

Step G: to a solution of compound 41-7 (0.288 g, 0.985 mmol, 1 eq.) in tetrahydrofuran (10 ml) was added N, N-carbonyldiimidazole (239.66 mg, 1.48 mmol, 1.5 eq.) and triethylamine (498.54 mg, 4.93 mmol, 5 eq.) and the mixture was stirred at 25 degrees celsius for 1 hour. Compound 7-3 (0.445 g, 1.97 mmol, 2 eq, hydrochloride) was added to the reaction solution and the mixture stirred at 25 degrees celsius for 12 hours. After completion of the reaction, the reaction mixture was quenched with water (30 ml), extracted with ethyl acetate (40 ml. Times.2), the organic phases were combined, washed with saturated brine (20 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a residue which was purified by high performance liquid chromatography (column: phenomenex luna C, 150 x 25 mm. 10 μm; mobile phase [ water (formic acid) -acetonitrile)]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 34% -64%,12 min) and then separated by SFC (column: DAICEL CHIRALCEL OJ (250 mm x 30 mm, 10 μm); mobile phase: [0.1% Ammonia Water methanol ]]The method comprises the steps of carrying out a first treatment on the surface of the Methanol%: 30% -30%,2.8 min) to give compound 41.MS (ESI) m/z 507.2[ M+H ] ⁺ ].

¹ H NMR(CD ₃ OD,400MHz):δppm 8.64(s,2H),7.91-7.80(m,1H),5.91-5.76(m,2H),4.62(br d,J＝4.8Hz,2H),4.44-4.28(m,1H),4.02-3.87(m,4H),3.58(br s,4H),1.84-1.62(m,2H),1.57-1.36(m,2H),1.00(t,J＝7.2Hz,3H).

Compound 41 was detected by SFC [ column model: chiralcel OJ-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, and phase B is methanol (0.05% diethylamine); gradient (B%): 5% -40%) to obtain: the retention time of compound 41 was 1.682min and the e.e. value was 100.000%.

Example 42

Step A: to a solution of compound 42-1 (10 g, 112.19 mmol, 1 eq.) in tetrahydrofuran (30 ml) and water (30 ml) was added potassium carbonate (31.01 g, 224.38 mmol, 2 eq.) and di-tert-butyl dicarbonate (25.71 g, 117.80 mmol, 27.06 ml, 1.05 eq.) and the mixture was stirred at 25 degrees celsius for 12 hours. The reaction solution was extracted with ethyl acetate (100 ml. Times.2), the organic phases were combined, washed with saturated brine (30 ml. Times.2), dried over anhydrous sodium sulfate, filtered and concentrated to give compound 42-2. ¹ H NMR (chloroform-d, 400 MHz): delta (ppm) 4.72-4.28 (m, 1H), 4.02-3.81 (m, 1H), 3.64 (dd, J=7.6, 2.8Hz, 2H), 3.05-2.57 (m, 1H), 1.94-1.72 (m, 1H), 1.46 (s, 9H), 1.40-1.29 (m, 1H), 1.20 (d, J=6.8 Hz, 3H).

And (B) step (B): to a solution of compound 42-2 (21.20 g, 112.02 mmol, 1 eq.) in dichloromethane (200 ml) at 0 degrees celsius was added dess-martin oxidant (46.56 g, 109.78 mmol, 33.99 ml, 0.98 eq.) and the mixture was stirred at 25 degrees celsius for 2 hours. The reaction mixture was quenched by adding a saturated sodium thiosulfate solution (100 ml), stirred for 10 minutes, extracted with ethyl acetate (300 ml), the organic phase was washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography (silica 1000 mesh silica gel, eluent gradient ethyl acetate 10/1 to 2/1) of ester/petroleum ether to give compound 42-3. ¹ H NMR (chloroform-d, 400 MHz): delta (ppm) 9.76 (s, 1H), 5.00-4.59 (m, 1H), 4.21-4.07 (m, 1H), 2.69-2.50 (m, 2H), 1.43 (s, 9H), 1.23 (d, J=6.8 Hz, 3H).

Step C: to a solution of compound 42-3 (19.45 g, 103.88 mmol, 1 eq.) and lithium chloride (13.21 g, 311.64 mmol, 6.38 ml, 3 eq.) in acetonitrile (100 ml) at 0 degrees celsius was added N, N-diisopropylethylamine (40.28 g, 311.64 mmol, 54.28 ml, 3 eq.) and the mixture was stirred at 0 degrees celsius for 30 minutes, then a solution of compound 1-13 (46.58 g, 207.76 mmol, 41.22 ml, 2 eq.) in acetonitrile (20 ml) was added at 0 degrees celsius and the mixture was naturally warmed to 25 degrees celsius and stirred for 12 hours. Hydrochloric acid (1 mol per liter, 250 ml) was added to the reaction solution, stirred at 25 ℃ for 2 minutes, extracted with ethyl acetate (380 ml), the organic phase was washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the residue was purified by column chromatography (silica 1000 mesh silica gel, eluent gradient ethyl acetate/petroleum ether 100/1 to 3/1) to give compound 42-4. ¹ H NMR (chloroform-d, 400 MHz): delta (ppm) 7.01-6.85 (m, 1H), 5.94-5.82 (m, 1H), 4.65-4.28 (m, 1H), 4.21 (d, J=6.4 Hz, 2H), 3.97-3.64 (m, 1H), 2.61-2.18 (m, 2H), 1.45 (s, 9H), 1.30 (s, 3H), 1.21-1.10 (m, 3H).

Step D: to a solution of compound 42-4 ((22.18 g, 86.19 mmol, 1 eq.) in tetrahydrofuran (200 ml) was added diisobutylaluminum hydride (1 mol per liter, 258.58 ml, 3 eq.) and the reaction solution was stirred at 0℃for 3 hours, aqueous solution of saturated sodium potassium tartrate (60 ml) was added dropwise thereto while keeping the temperature at 0℃and then naturally warmed to 25℃and stirred for 12 hours, the mixture was extracted with ethyl acetate (200 ml), the organic phase was washed with saturated brine (50 ml. Times.2), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography (silica 1000 mesh silica gel, eluent gradient ethyl acetate/petroleum ether 100/1 to 2/1) to give compound 42-5. ¹ H NMR (chloroform-d, 400 MHz) delta (ppm) 5.83-5.57 (m, 2H), 4.54-4.22 (m, 1H), 4.12 (d, J=4.4 Hz, 2H), 3.85-3.56 (m, 1H), 2.21 (s, 2H), 1.46(s,9H),1.14(d,J＝6.8Hz,3H).

Step E: to a solution of compound 42-5 (5.63 g, 26.15 mmol, 1 eq.) in dichloromethane (40 ml) at 0 degrees celsius was added triphenylphosphine (9.7 g, 36.98 mmol, 1.41 eq.) and carbon tetrabromide (12.23 g, 36.87 mmol, 1.41 eq.) and the mixture was reacted at 0 degrees celsius for 2 hours. The reaction solution was concentrated, and the residue was purified by column chromatography (1000 mesh silica gel, petroleum ether/ethyl acetate=1000/1, 10/1) to give compound 42-6. ¹ H NMR (chloroform-d, 400 MHz) delta (ppm) 5.87-5.69 (m, 2H), 4.58-4.20 (m, 1H), 4.03-3.93 (m, 2H), 3.86-3.62 (m, 1H), 2.39-2.13 (m, 2H), 1.46 (s, 9H), 1.14 (d, J=6.8 Hz, 3H).

Step F: to a solution of compound 42-6 (2 g, 7.19 mmol, 1 eq.) in dimethyl sulfoxide (20 ml) was added sodium cyanide (0.63 g, 12.86 mmol, 1.79 eq.) and the reaction was heated to 100 degrees celsius for 5 hours. The reaction solution was cooled to 25 degrees celsius, water (50 ml) was added, and stirred for 5 minutes. Extraction with ethyl acetate (80 ml×2), washing the combined organic phases with saturated brine (30 ml×2), drying over anhydrous sodium sulfate, filtration, concentration, and purification of the resulting residue by column chromatography (1000 mesh silica gel, petroleum ether/ethyl acetate=100/1, 5/1) afforded compound 42-7. ¹ H NMR (chloroform-d, 400 MHz): delta ppm 4.12-3.85 (m, 2H), 2.93 (br s, 2H), 2.14-1.89 (m, 2H), 1.80-1.61 (m, 1H), 1.50 (s, 9H), 1.12-1.32 (m, 3H).

Step G: compound 42-7 (350 mg, 1.56 mmol, 1 eq.) was added to methanol hydrochloride (4 mol per liter, 10 ml) and stirred at 25 degrees celsius for 2 hours. The reaction solution was concentrated to obtain compound 42-8.

Step H: compound 42-8 (0.302 g, 1.56 mmol, 1 eq, hydrochloride salt), 1-7 (496.93 mg, 1.56 mmol, 1 eq) and sodium bicarbonate (524.00 mg, 6.24 mmol, 242.59 μl, 4 eq) were added to acetonitrile (10 ml) and the mixture was stirred at 25 degrees celsius for 12 hours. Water (20 ml) was added to the reaction solution, stirred for 2 minutes, the aqueous phase was extracted with ethyl acetate (80 ml), washed with saturated brine (30 ml. Times.2), dried over anhydrous sodium sulfate, filtered, and concentrated to give a residue, which was passed through Purification by thin layer chromatography (petroleum ether/ethyl acetate=3/1) afforded compound 42-9.MS (ESI) m/z 440.1[ M+H ] ⁺ ].

Step I: to a solution of compound 42-9 (0.173 g, 393.69 micromoles, 1 eq.) in trifluoroacetic acid (3 ml) was added trifluoromethanesulfonic acid (0.6 ml) and the reaction stirred at 25 degrees celsius for 2 hours. The reaction solution was concentrated to obtain a crude product of compound 42-10.MS (ESI) m/z 320.0[ M+H ] ⁺ ].

Step J: to a solution of compounds 42-10 (125 mg, 391.51 μmol, 1 eq.) in methanol (5 ml) and water (5 ml) was added lithium hydroxide (93.76 mg, 3.92 mmol, 10 eq.) and the reaction stirred at 25 degrees celsius for 12 hours. The reaction solution was adjusted to pH 3 with hydrochloric acid, extracted with ethyl acetate (80 ml), washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 42-11.MS (ESI) m/z 306.1[ M+H ] ⁺ ].

Step K: to a solution of compound 42-11 (0.12 g, 393.121. Mu. Mol, 1 eq.) in N, N-dimethylformamide (3 ml) were added O- (7-azabenzotriazol-1-yl) N, N, N, N-tetramethylurea hexafluorophosphate (298.95 mg, 786.23. Mu. Mol, 2 eq.) and N, N-diisopropylethylamine (254.04 mg, 1.97. Mu. Mol, 342.37. Mu. L, 5 eq.) followed by compound 7-3 (148.77 mg, 786.23. Mu. Mol, 2 eq.) and the reaction was stirred at 25℃for 12 hours. Hydrochloric acid (20 ml, 1 mol/l) was added to the reaction solution, stirred for 5 minutes, the aqueous phase was extracted with ethyl acetate (80 ml. Times.2), the organic phases were combined, washed with saturated brine (20 ml. Times.2), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by high performance liquid chromatography (column: phenomenex luna C; 150 x 25 mm. 10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 27% -57%,10 min) to give compound 42.MS (ESI) m/z 477.2[ M+H ] ⁺ ]. ¹ H NMR (chloroform-d, 400 MHz) delta (ppm) 8.59-8.52 (m, 2H), 7.78-7.70 (m, 1H), 5.58-5.51 (m, 2H), 4.77-4.65 (m, 1H), 3.98-3.79 (m, 4H), 3.76-3.52 (m, 4H), 2.63-2.41 (m, 2H), 2.40-2.26 (m, 2H), 1.39-1.37 (m, 3H).

Example 43

Step A: to a solution of compound 11-8 (300 mg, 1.14 mmol, 1 eq.) in dichloromethane (6 ml) was added N-methylmorpholine (172.93 mg, 1.71 mmol, 187.97 μl, 1.5 eq.) and compound 12-4 (275.68 mg, 1.37 mmol, 1.2 eq.) and the mixture was stirred at 25 ℃ for 12 hours. The reaction solution was diluted with water (30 ml), extracted with ethyl acetate (30 ml×2), the organic phases were combined, washed with saturated brine (30 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the crude product was purified by column chromatography (silica gel column, petroleum ether/ethyl acetate=3/1, 1/1, 0/1) to give compound 43-1.MS (ESI) m/z 429.1[ M+H ] ⁺ ].

Step A: to a solution of compound 43-2 (2 g, 9.33 mmol, 1 eq.) in N, N-dimethylformamide (50 ml) was added potassium carbonate (1.93 g, 14.00 mmol, 1.5 eq.) and compound 7-1 (1.82 g, 13.07 mmol, 1.4 eq.) and the mixture was stirred at 80 degrees celsius for 12 hours. The reaction solution was cooled to 25℃and water (200 ml) was added, and a white solid precipitated, filtered, and the cake was collected and concentrated under reduced pressure to give Compound 43-3.MS (ESI) m/z 218.2[ M+H ] ⁺ -100].

And (B) step (B): to a solution of compound 43-3 (2.4 g, 7.56 mmol, 1 eq.) in ethyl acetate (25 ml) was added ethyl acetate hydrochloride (4 mol per liter, 25 ml, 13.22 eq.) and the reaction was stirred at 25 degrees celsius for 1 hour. The reaction mixture was concentrated to give the hydrochloride salt of compound 43-4. MS (ESI) m/z 218.1[ M+H ] ⁺ ].

Step C: to compound 43-1 (1.7 g, 3.97 mmol, 1 eq.) and compound 43-4 (1.31 g, 5.16 mmol, 1.3 eq., hydrochloride)To a solution of N, N-dimethylformamide (30 ml) was added triethylamine (602.43 mg, 5.95 mmol, 828.65 μl, 1.5 eq.) and the mixture was stirred at 25 degrees celsius for 12 hours. The reaction solution was diluted with water (100 ml), extracted with ethyl acetate (100 ml×2), the organic phases were combined, washed with saturated brine (100 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the resulting crude product was purified by column chromatography (silica gel column, eluent gradient 0 to 100% petroleum ether/ethyl acetate, 80 ml/min) to give compound 43.MS (ESI) m/z 507.2[ M+H ] ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.52(br s,1H),8.74(s,2H),7.82(s,1H),6.49(br d,J＝3.6Hz,1H),5.86-5.65(m,2H),4.82(br d,J＝12.8Hz,2H),4.52(br s,3H),4.14(br s,1H),3.01(br s,2H),2.68(s,3H),1.70-1.55(m,4H),1.31(br d,J＝6.4Hz,3H)

Compound 43 was detected by SFC [ column model: chiralpak IC-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is methanol+acetonitrile (0.05% diethylamine); gradient (B%): 50% -50%) to obtain: the retention time of compound 43 was 0.893min and the e.e. value was 95.318%.

Example 44

Step A: to a solution of compound 44-1 (300 mg, 1.61 mmol, 1 eq.) and compound 7-1 (269.72 mg, 1.93 mmol, 1.2 eq.) in N, N-dimethylformamide (5 ml) was added potassium carbonate (333.92 mg, 2.42 mmol, 1.5 eq.) and the mixture was stirred at 90 degrees celsius for 2 hours. The reaction solution was cooled to 25℃and water (50 ml) was added, and a white solid precipitated, filtered, and the cake was collected and concentrated under reduced pressure to give Compound 44-2.MS (ESI) m/z 234.1[ M+H ] ⁺ -56].

And (B) step (B): to a solution of compound 44-2 (155 mg, 535.72 micromoles, 1 eq.) in ethyl acetate (2 ml) was added ethyl acetate hydrochloride (4 moles per liter,2 ml, 14.93 eq.) and the reaction stirred at 25 degrees celsius for 2 hours. The reaction mixture was concentrated to give hydrochloride of compound 44-3. MS (ESI) m/z 190.1[ M+H ] ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δ＝9.86(br s,2H),8.79(s,2H),4.38(br s,2H),4.31-4.24(m,2H),4.16-4.06(m,1H),2.55(t,J＝5.2Hz,3H).

Step C: to a solution of compound 43-1 (158.16 mg, 369.26 μmol, 1 eq) and compound 44-3 (100 mg, 443.11 μmol, 1.2 eq, hydrochloride) in N, N-dimethylformamide (5 ml) was added triethylamine (56.05 mg, 553.89 μmol, 77.09 μmol, 1.5 eq) and the mixture was stirred at 25 degrees celsius for 12 hours. The reaction solution was diluted with water (50 ml), extracted with ethyl acetate (50 ml. Times.2), the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product which was purified by high performance liquid chromatography (column: phenomenex luna C, 150 x 25 mm. Times.10 μm; mobile phase: [ water (formic acid) -acetonitrile) ]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 22% -52%,2 min), high performance liquid chromatography (column: waters Xbridge 150 x 25 mm x 5 microns; mobile phase [ water (ammonium bicarbonate) -acetonitrile]The method comprises the steps of carrying out a first treatment on the surface of the Acetonitrile%: 25% -55%,8 min) to give compound 44.MS (ESI) m/z 479.2[ M+H ] ⁺ ].

¹ H NMR(400MHz,DMSO-d ₆ )δ＝12.51(br s,1H),8.74(s,2H),7.82(s,1H),6.50(br d,J＝3.6Hz,1H),5.92-5.68(m,2H),4.98(br s,1H),4.53(d,J＝5.2Hz,2H),4.37-4.17(m,4H),2.93(s,3H),2.07(s,1H),1.30(d,J＝6.4Hz,3H)

Compound 44 was detected by SFC [ column model: chiralpak IC-3.50X4.6mm I.D.,3 μm; mobile phase: phase A is supercritical carbon dioxide, phase B is methanol+acetonitrile (0.05% diethylamine); gradient (B%): 40% -40%) to obtain: the retention time of compound 44 was 1.340min and the e.e. value was 100.000%.

Experimental example 1: PARP7 enzyme Activity test

The purpose of the experiment is as follows:

this assay was used to examine the efficacy of compounds to inhibit PARP7 enzyme activity, with lower IC ₅₀ The values represent the high potency of the compounds as PARP7 inhibitors in the following assay settings.

Experimental materials:

PARP7 chemofluorescence detection kit was purchased from BPS Bioscience.

The experimental method comprises the following steps:

preparing PBST buffer solution: 1 times PBS containing 0.05% Tween-20, i.e., 10mL PBS with 5. Mu.L 100% Tween-20

Diluting the histone solution in the kit by 5 times with 1 time PBS, taking 25 mu L/hole diluent into a micro-pore plate, and standing at 4 ℃ for overnight incubation; after the incubation is finished, discarding the liquid in the wells, taking 100 mu L/well PBST wash plate for 3 times, discarding the residual liquid in the wells; taking 100 mu L/hole sealing liquid into a micro-pore plate, and placing the micro-pore plate at 25 ℃ for incubation for 90 minutes; after the incubation is finished, discarding the liquid in the wells, taking 100 mu L/well PBST wash plate for 3 times, discarding the residual liquid in the wells;

1 time test buffer preparation: 10 times of PARP test buffer solution is diluted by 10 times by double distilled water;

compound solution preparation: test compounds were diluted 5-fold to 8 th concentration, i.e. from 1000 μm to 12.8nM, with 100% dmso. And then 1-time test buffer solution is used for diluting each gradient of the compound to be tested into working solution with DMSO of 10%, 2.5 mu L/hole is taken and added into a micro-pore plate, and a double-multi-hole experiment is set.

12.5. Mu.L/well bottom mix (1.25. Mu.L 10 fold PARP test buffer; 1.25. Mu.L 10 fold PARP test mix; 10. Mu.L double distilled water) was applied to the microplate.

PARP7 enzyme was diluted to 6 ng/. Mu.L and 10. Mu.L/well was added to the microplate, at which point the final concentration gradient of the compound was 10. Mu.M to 0.128nM, PARP7 (60 ng/well) and the reaction system incubated at 25℃for 60 minutes;

after the incubation is finished, discarding the liquid in the wells, taking 100 mu L/well PBST wash plate for 3 times, discarding the residual liquid in the wells;

50-fold dilution of strepavidin-HRP with blocking solution, and then taking 25 mu L/well to the microplate, and incubating at 25 ℃ for 30 minutes;

after the incubation is finished, discarding the liquid in the wells, taking 100 mu L/well PBST wash plate for 3 times, discarding the residual liquid in the wells;

ELISA ECL substrate A and ELISA ECL substrate B were mixed 1:1 (v/v) and 50. Mu.L/well was applied to the microplate and chemiluminescent values were read.

Experimental results: see table 1.

TABLE 1 inhibition of PARP7 enzyme Activity

Sample of	PARP7 IC 50 (nM)
Compound 1	57.76
Compound 2	10.98
Compound 3	10.42
Compound 4	24.94
Compound 5	77.13
Compound 6	53.76
Compound 7	75.69
Compound 8	36.38
Compound 10	29.78
Compound 11	7.97

Compound 12	4.54
Compound 13	9.15
Compound 14	8.68
Compound 15	9.18
Compound 16	32.00
Compound 17	5.81
Compound 18	13.35
Compound 19	9.37
Compound 20	11.27
Compound 21	5.35
Compound 22	8.38
Compound 23	21.12
Compound 24	5.05
Compound 25	11.68
Compound 27	18.34
Compound 28A	7.32
Compound 28B	7.13
Compound 29A	7.41
Compound 29B	6.64
Compound 30	4.91
Compound 31	4.89
Compound 32	6.06
Compound 33	6.10
Compound 34	7.18
Compound 37	13.81
Compound 43	8.65

Conclusion: the compounds of the present invention have excellent in vitro inhibitory activity against PARP 7.

Experimental example 2: h1373 cell antiproliferative assay

Experimental materials:

RPMI-1640 medium, penicillin/streptomycin antibiotics were purchased from Vison's Tex, and fetal bovine serum from Biosera. CellTiter-Glo (cell viability chemiluminescent detection reagent) reagent was purchased from Promega. The NCI-H1373 cell line was purchased from Nanjac Bai Biotechnology Co. Nivo Multi-labelling Analyzer (Perkinelmer).

The experimental method comprises the following steps:

h1373 cells were seeded in white 96-well plates, 80 μl of cell suspension per well, containing 3000H 1373 cells. Cell plates were placed in a carbon dioxide incubator overnight for culture.

The test compounds were diluted 5-fold to the 9 th concentration, i.e. from 2mM to 5.12nM, using a row gun and a double multiplex well experiment was set up. 78. Mu.L of medium was added to the intermediate plate, and 2. Mu.L of the gradient diluted compound per well was transferred to the intermediate plate at the corresponding position, and 20. Mu.L of the gradient diluted compound per well was transferred to the cell plate after mixing. The concentration of compound transferred into the cell plate ranged from 10. Mu.M to 0.0256nM. The cell plates were placed in a carbon dioxide incubator for 6 days. A further cell plate was prepared and signal values read on the day of dosing as maximum values were used in the data analysis. To this plate, 25. Mu.L of cell viability chemiluminescent detection reagent was added per well and incubated at room temperature for 10 minutes to stabilize the luminescent signal. Multiple marker analyzer readings were used.

The luminescence signal was stabilized by adding 25. Mu.L of the cell viability chemiluminescent detection reagent per well to the cell plate 6 days after administration and incubating at room temperature for 10 minutes. Multiple marker analyzer readings were used.

Data analysis:

conversion of raw data to inhibition ratio, IC ₅₀ The values of (2) can be obtained by curve fitting with four parameters. Table 2 provides the inhibitory activity of the compounds of the invention on H1373 cell proliferation.

Experimental results: see table 2:

TABLE 2 inhibitory Activity of the Compounds of the invention on H1373 cell proliferation

Sample of	H1373 antiproliferative IC 50 (nanomoles per liter)
Compound 1	5.06
Compound 2	12.52
Compound 4	73.14
Compound 6	10.35
Compound 7	33.53
Compound 8	66.16
Compound 9	57.74
Compound 10	14.50
Compound 11	83.01
Compound 12	105.1
Compound 14	69.86
Compound 20	23.05
Compound 21	158.2
Compound 24	16.23
Compound 25	71.64

Compound 28B	72.95
Compound 31	11.81
Compound 34	22.81

Conclusion: the compounds of the present invention have excellent H1373 antiproliferative activity.

Experimental example 3: CT26 CXCL10 mRNA activation assay

Experimental materials:

RPMI-1640 medium was purchased from Viva cell, fetal bovine serum from Biosera, RNA extraction, reverse transcription and qPCR reagents from Norflua. CT26 cell line was purchased from cell bank of China academy of sciences. Quantitive fluorescence of quantsuio 6 PCR (Thermo fisher).

The experimental method comprises the following steps:

CT26 cells were seeded in 24-well plates, 400. Mu.L of cell suspension per well, containing 200000 CT26 cells. Cell plates were placed in a carbon dioxide incubator overnight for culture.

The test compounds were diluted 3-fold to the 8 th concentration, i.e., from 200. Mu.M to 914.5nM, using a row gun. 156. Mu.L of medium was added to the intermediate plate, and then 4. Mu.L of the gradient diluted compound per well was transferred to the intermediate plate at the corresponding position, and 100. Mu.L of the gradient diluted compound per well was transferred to the cell plate after mixing. The concentration of compound transferred into the cell plate ranged from 10. Mu.M to 4.57nM. The cell plates were placed in a carbon dioxide incubator for 24 hours.

The medium in the cell plate was removed, 500. Mu.L of RNA extraction reagent per well was added to the cell plate, and RNA was extracted according to the protocol. Reverse transcription was performed using 1. Mu.g RNA to obtain cDNA, cDNA solution according to 1:4 after dilution, 2. Mu.L of each well was added to the qPCR reaction.

Data analysis:

conversion of the original CT value data into relative expression data, EC ₅₀ The values of (2) can be obtained by curve fitting with four parameters. Table 3 providesThe induction activity of the compounds of the invention on CT26 CXCL10mRNA expression is shown.

Experimental results: see table 3:

TABLE 3 mRNA expression-inducing Activity of the compounds of the invention on CXCL10 in CT26 cells

Sample of	CXCL10 EC 50 (nanomole per liter)
Compound 2	299.9
Compound 7	14.75
Compound 8	22.33
Compound 14	162.7
Compound 22	329
Compound 24	97.71
Compound 25	389.1
Compound 30	114.1
Compound 31	28.97

Compound 34

75.15

Conclusion: the compounds of the invention are capable of inducing CXCL10mRNA expression in CT26 cells.

Test example 4: PARP1 enzyme Activity test experiment

Experimental materials:

PARP1 chemofluorescence detection kit was purchased from BPS Bioscience; enVision Multi-tag Analyzer (Perkinelmer).

The experimental method comprises the following steps:

1. and (3) preparation of a reagent:

preparing PBST buffer solution: 1 XPBS containing 0.05% Tween-20, i.e., 10mL PBS with 5. Mu.L 100% Tween-20

1X test buffer preparation: 10 XPARP test buffer was diluted 10-fold with double distilled water

2. Compound preparation:

compound solution preparation: test compounds were diluted 4-fold to 8 th concentration, i.e. from 200 μm to 120nM, with 100% dmso. The test compounds were then serially diluted with 1X assay buffer to working solution with 10% DMSO.

3. Experimental operation:

diluting the histone solution in the kit by 5 times with 1 XPBS, taking 25 mu L/hole diluent into a micro-pore plate, and standing at 4 ℃ for overnight incubation;

after the incubation is finished, discarding the liquid in the wells, taking 100 mu L/well PBST wash plate for 3 times, discarding the residual liquid in the wells;

taking 100 mu L/hole sealing liquid into a micro-pore plate, and placing the micro-pore plate at 25 ℃ for incubation for 90 minutes; after the incubation is finished, discarding the liquid in the wells, taking 100 mu L/well PBST wash plate for 3 times, discarding the residual liquid in the wells;

12.5. Mu.L/well bottom mix (1.25. Mu.L 10 XPARP test buffer; 1.25. Mu.L 10 XPARP test mix; 2.5. Mu.L Activated DNA; 7.5. Mu.L double distilled water) was applied to the microplate.

2.5 mu L/well of compound working solution was applied to the microplate and a double-well experiment was performed.

Diluting PARP1 enzyme to 2 ng/mu L, adding 10 mu L/hole to a micro-pore plate, wherein the final concentration gradient of the compound to be tested is 2 mu M to 0.12nM, and incubating the reaction system at 25 ℃ for 60 minutes;

After the incubation is finished, discarding the liquid in the wells, taking 100 mu L/well PBST wash plate for 3 times, discarding the residual liquid in the wells;

50-fold dilution of strepavidin-HRP with blocking solution, and then taking 25 mu L/well to the microplate, and incubating at 25 ℃ for 30 minutes;

after the incubation is finished, discarding the liquid in the wells, taking 100 mu L/well PBST wash plate for 3 times, discarding the residual liquid in the wells;

ELISA ECL substrate A and ELISA ECL substrate B were mixed 1:1 (v/v) and 50. Mu.L/well was applied to the microplate and chemiluminescent values were read.

Data analysis

Converting the raw data into enzyme activity, IC ₅₀ The values of (2) can be obtained by curve fitting with four parameters.

Experimental results: see table 4:

TABLE 4 inhibition of PARP1 enzyme Activity by the Compounds of the invention

Sample of	PARP1 IC 50 (nanomoles per liter)
Compound 31	3635
Compound 34	4308

Compound 39	6079
Compound 40	254.9
Compound 41	221.3

Conclusion: the compounds of the present invention have high selectivity for PARP 1.

Experimental example 5: MDA-MB-436 cell antiproliferative assay

Experimental materials:

RPMI-1640 medium, fetal bovine serum, penicillin/streptomycin antibiotics were purchased from Vison. MDA-MB-436 cell line was purchased from the cell bank of the national academy of sciences. Envision multi-label analyzer (PerkinElmer).

The experimental method comprises the following steps:

MDA-MB-436 cells were seeded in a white 96-well plate, 80. Mu.L of cell suspension per well, containing 3000 MDA-MB-436 cells. Cell plates were placed in a carbon dioxide incubator overnight for culture.

The test compounds were diluted 5-fold to the 8 th concentration, i.e. from 2mM to 26nM, using a row gun and a double multiplex well experiment was set up. 78. Mu.L of medium was added to the intermediate plate, and 2. Mu.L of the gradient diluted compound per well was transferred to the intermediate plate at the corresponding position, and 20. Mu.L of the gradient diluted compound per well was transferred to the cell plate after mixing. The cell plates were placed in a carbon dioxide incubator for 7 days. A cell plate was also prepared and the signal values read on the day of dosing as Max values to participate in the data analysis. To this plate, 25 mu L Promega CellTiter-Glo was added per well and incubated at room temperature for 10 minutes to stabilize the luminescence signal. A PerkinElmer Envision multi-label analyzer reading was used.

To the cell plate 25. Mu.L of Promega CellTiter-Glo reagent per well was added and incubated at room temperature for 10 minutes to stabilize the luminescence signal. A PerkinElmer Envision multi-label analyzer reading was used.

Data analysis:

conversion of raw data to inhibition ratio, IC ₅₀ The values of (2) can be obtained by curve fitting with four parameters. Table 6 provides the inhibitory activity of the compounds of the invention on MDA-MB-436 cell proliferation.

Experimental results: see table 5:

TABLE 5 inhibitory Activity of the Compounds of the invention against MDA-MB-436 cell proliferation

Sample of	MDA-MB-436 antiproliferative IC 50 (nanomoles per liter)
Compound 2	2096
Compound 31	2675
Compound 34	2739

Conclusion: the compounds of the invention have no obvious inhibition effect on MDA-MB-436 antiproliferative activity.

Experimental example 6: PARYlation cell experiment

Experimental materials:

DMEM medium was purchased from Viva cell. Hela cells were purchased from Nanjac, bai. Poly/Mono-ADP Ribose (E6F 6A) Rabbit mAb was purchased from CST. Fluorescein (FITC) -conjugated Affinipure Goat Anti-Rabbit IgG (H+L) was purchased from Wuhan Sanying. Hydrogen peroxide was purchased from Alatine. DAPI was purchased from aladine. PBS was purchased from vitamin. Methanol is purchased from chinese medicine. Acetone was purchased from chinese medicine. Tween-20 was purchased from a national drug. Skim milk powder was purchased from illi. Envision multi-label analyzer (PerkinElmer).

The experimental method comprises the following steps:

h1373 cells were seeded in white 96-well plates, 80 μl of cell suspension per well, containing 3000H 1373 cells. Cell plates were placed in a carbon dioxide incubator overnight for culture.

The first day:

hela cells were plated at 60000 cells/well, 37 degrees, 5% CO ₂ Overnight.

The following day:

and (3) preparation of a reagent:

washing liquid: tween-20 was added to 1 XPBS, and the final concentration of Tween-20 was 0.05%.

Sealing liquid: and adding skimmed milk powder into the washing liquid, wherein the final concentration of the skimmed milk powder is 5%.

Cell fixative: methanol and acetone were mixed in a 7:3 ratio.

Preparing a compound to be tested:

the test compounds were diluted 5-fold to the 8 th concentration, i.e. from 1mM to 13nM, with a DMSO concentration of 100% using a row gun.

mu.L of each concentration gradient of the compound was taken into the compound intermediate plate 1, and 99. Mu.L of PBS was added to mix well, at which time the final concentration of the compound was 10. Mu.M to 0.13nM and the DMSO concentration was 1%. Double complex holes are arranged.

mu.L of each concentration gradient of the compound is taken into a compound middle plate 2, 99 mu.L of PBS containing 50mM hydrogen peroxide is added and mixed for standby, and the final concentration of the compound is 10 mu M to 0.13nM at the moment, and the concentration of DMSO is 1%. Double complex holes are arranged.

The experimental steps are as follows:

1. the supernatant was removed from the cells and 40. Mu.L of compound per well was taken from the compound intermediate plate 1 to the cell plate and incubated at 37℃for 30 minutes. The requirements include:

compound wells: the corresponding compound was added, DMSO 1%

Negative control and positive control: 1% DMSO was added

Blank control: cell-free wells, PBS was added

2. After the incubation was completed, 40. Mu.L of compound per well was taken from compound intermediate plate 2 to the cell plate with a final H2O2 concentration of 0.5mM.

Compound wells: compound +1mM H ₂ O ₂

Positive control and negative control: 1% DMSO+1mM H ₂ O ₂

Blank control: cell-free wells, PBS was added

Incubate at 3.25 degrees for 10 minutes.

4. After the incubation was completed, the wells were washed 1 time with ice-chilled PBS, 100. Mu.L of pre-chilled cell fixative was added to each well, left at-20℃for 10 minutes, and then the cell fixative was thrown away.

5. After air-conditioning the stem cell plates, 200 μl of PBS per well was added for washing and the PBS was discarded.

6. 100. Mu.L of blocking solution per well was added and incubated at 25℃for 30 minutes, after which the blocking solution was thrown off.

7. mu.L Poly/Mono-ADP Ribose (E6F 6A) Rabbit mAb per well was added and diluted 1:500 in blocking solution and incubated at 25℃for 60 min.

Negative control wells: adding 25. Mu.L/well of blocking solution

Blank control wells: adding 25. Mu.L/well of blocking solution

8. After the incubation was completed, the cell plates were washed 4 times with 200. Mu.L of wash solution per well for 3 minutes, and the wash solution was thrown off.

9. Add 25. Mu.L of blocking solution per well containing 1:200 dilution Fluorescein (FITC) -conjugated Affinipure Goat Anti-Rabbit IgG (H+L) and 0.5. Mu.g/mL DAPI,25℃incubation for 60 min.

10. After the incubation was completed, the cell plates were washed 4 times with 200 μl of wash solution per well for 3 minutes each.

11. After removal of the liquid, the corresponding fluorescence values were read on Envision: FITC: accounting 480nm,Emission 530nm; DAPI: and accounting 360nm,Emission 460nm.

Data analysis:

carrying out data normalization processing on the original data, converting the normalized data into inhibition rate and IC ₅₀ The values of (2) can be obtained by curve fitting with four parameters. Table 6 provides the inhibitory activity of the compounds of the present invention.

Experimental results: see table 6:

TABLE 6 inhibition of Hela cell PARYstation by the compounds of the invention

Sample of	PARYLATION IC 50 (nanomole per liter)
Compound 2	8278
Compound 31	2818
Compound 34	>10000

Conclusion: the compound has weak inhibition effect on PARYstation in Hela cells and good selectivity.

Experimental example 7: in vivo pharmacokinetic studies in mice

The purpose of the experiment is as follows:

the aim of this experiment was to evaluate the pharmacokinetic behavior of the compounds following a single intravenous injection and intragastric administration and to investigate the bioavailability following intragastric administration.

Experimental operation:

CD-1 male mice 7 to 10 weeks old were selected. Mice were fasted for at least 12 hours prior to dosing and fed was resumed 4 hours after dosing, with free water throughout the trial period.

The animals of the intravenous group on the day of the experiment are given the corresponding compounds by single injection through the tail vein, and the administration volume is 5 milliliters per kilogram; animals in the oral group were given the corresponding compound by single gavage in a volume of 10 ml/kg. Animals were weighed prior to dosing and dosing volumes were calculated from body weight. Sample collection time: the injection groups were 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 24 hours, and the gavage groups were 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 24 hours. About 30 μl of whole blood was collected through saphenous vein at each time point for preparing plasma for concentration determination by high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). All animals were CO-processed after collection of PK samples at the last time point ₂ The anesthesia is euthanized. By WinNonlin ^TM Non-compartmental model of Version 6.3 (Pharsight, mountain View, CA) pharmacokinetic software plasma concentrations were processed and pharmacokinetic parameters were calculated using the linear log trapezium method.

Experimental results: the PK profile evaluation results are shown in table 7.

Conclusion of experiment:

the compounds of the present invention have excellent pharmacokinetic properties.

TABLE 7 evaluation results of PK Properties in vivo

Conclusion of experiment:

the compound of the invention has excellent AUC and bioavailability in vivo in mice and good pharmacokinetic property.

Claims (16)

1. A compound of formula (IV) or a pharmaceutically acceptable salt thereof

   Wherein,

   ring A is selected from 4-10 membered heterocycloalkyl and C _4-10 Cycloalkyl, the 4-10 membered heterocycloalkyl and C _4-10 Cycloalkyl groups are each independently optionally substituted with 1, 2 or 3R _a Substitution;

   R _a are each independently selected from H, CN, OH, F, cl, br, -COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy, said-COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino and C _1-3 Alkoxy groups are each independently optionally substituted with 1, 2 or 3R;

   ring B is selected from 5-6 membered heteroaryl;

   L ₅ selected from single bonds, -O-, -S-, and-NR ₁ -；

   R ₁ Are independently selected from H, D, -C _1-3 alkyl-OH, -C _1-3 alkyl-NH ₂ And C _1-3 Alkyl, said-C _1-3 alkyl-OH, -C _1-3 alkyl-NH ₂ And C _1-3 Alkyl is optionally substituted with 1, 2 or 3R _b Substitution;

   R _b are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

   L ₂ and L ₄ Are independently selected from single bond, -CH ₂ -、-CH ₂ -CH ₂ -and-CH ₂ -CH ₂ -CH ₂ -, the-CH ₂ -、-CH ₂ -CH ₂ -and-CH ₂ -CH ₂ -CH ₂ -each independently optionally being substituted with 1 or 2R _c Substitution;

   R _c are respectively and independently selected from H, F, cl, br, D, C _1-3 Alkyl, -O-C _1-3 Alkyl and-C _1-3 alkyl-O-C _1-3 Alkyl, said C _1-3 Alkyl is each independently optionally substituted with 1, 2 or 3R;

   R _2a and R is _2b Are independently selected from H, F, cl, br, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

   L ₁ selected from single bond, -CH ₂ -、-O-、-C(＝O)-、-C(＝O)-NR ₄ -and-NR ₄ -；

   L ₃ Selected from single bond, -CH ₂ -、-O-、-C(＝O)-、-C(＝O)-NR ₄ -and-NR ₄ -；

   R ₄ Selected from H, D and C _1-3 Alkyl, said C _1-3 Alkyl is optionally substituted with 1, 2 or 3R;

   R ₃ are each independently selected from H, CN, OH, F, cl, br, -COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino, C _1-3 Alkoxy and C _3-6 Cycloalkyl, said-COOH, -CONH ₂ 、C _1-3 Alkyl, C _1-3 Alkylamino, C _1-3 Alkoxy and C _3-6 Cycloalkyl groups are each independently optionally substituted with 1, 2 or 3R;

   each R is independently selected from D, OH, CN, F, cl and Br;

   n is selected from 1, 2 and 3;

   the 4-10 membered heterocycloalkyl groups each independently contain 1, 2 or 3 heteroatoms or groups of heteroatoms selected from N, O, S and NH.
2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of

   Wherein n, ring A, ring B, L ₁ 、L ₂ 、L ₃ 、L ₄ 、L ₅ 、R _a 、R _2a 、R _2b And R is ₃ As defined in claim 1.
3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of

   Wherein,

   x is-O-, -CH ₂ -, -NH-or-N (CH) ₃ )-；

   Y is N, Z is CH; or Y is CH, Z is N;

   ring C is selected from 4-10 membered heterocycloalkyl, said 4-10 membered heterocycloalkylHeterocycloalkyl is optionally substituted with 1, 2 or 3R _a Substitution;

   p is 1 or 2;

   n, ring A, ring B, L ₁ 、L ₂ 、L ₃ 、R _2a 、R _2b 、Rc、R ₃ As defined in claim 1;

   the carbon atoms with "×" are chiral carbon atoms, either in the form of (R) or (S) single enantiomers or enriched in one enantiomer.
4. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein ring a is selected from azetidinyl, piperidinyl, piperazinyl, 1, 4-diazacyclohexanyl, 3, 8-diazabicyclo [3.2.1]Octyl and 2, 5-diazabicyclo [2.2.2]Octyl said azetidinyl, piperidinyl, piperazinyl, 1, 4-diazacyclohexenyl, 3, 8-diazabicyclo [3.2.1]Octyl and 2, 5-diazabicyclo [2.2.2]Octyl radicals are each independently optionally substituted with 1, 2 or 3R _a And (3) substitution.
5. A compound according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from pyrimidinyl, pyridinyl, pyridazinyl, pyrazolyl, thiophenyl and thiazolyl.
6. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein L ₅ Selected from-NH-.
7. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein L ₄ Selected from-CH (CH) ₃ )-、-CH(CH ₂ OCH ₃ )-、-CH(CH ₃ )CH ₂ CH ₂ -、-CH(CH ₃ )CH ₂ -、-CH(CH ₂ CH ₂ CH ₃ )-、-CH(CH ₂ CH ₂ OCH ₃ ) -and-CH (CH) ₂ OH)-。
8. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein R _2a And R is _2b Each independently selected from H and F.
9. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein L ₁ selecting-C (=O) -CH ₂ -, -C (=O) -NH-and-C (=O) -N (CH) ₃ )-。
10. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein L ₃ Selected from single bond, -CH ₂ -, -O-, -NH-and-N (CH) ₃ )-。
11. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein L ₂ Selected from-CH ₂ -and a single bond.
12. A compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein the structural unitSelected from the group consisting of
13. A compound according to claim 1 or 3 or a pharmaceutical thereofSalts which are acceptable in the above, wherein the structural unitsSelected from the group consisting of
14. A compound of the formula:
15. the compound according to claim 14, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of
16. Use of a compound according to any one of claims 1-15, or a pharmaceutically acceptable salt thereof, for the preparation of a PARP7 inhibitor.