CN117285517A

CN117285517A - Pyrimidinone derivative and pharmaceutical composition, preparation method and application thereof

Info

Publication number: CN117285517A
Application number: CN202310743290.8A
Authority: CN
Inventors: 栾林波; 姚元山; 陈永凯
Original assignee: Shanghai Meiyue Biotech Development Co Ltd
Current assignee: Shanghai Meiyue Biotech Development Co Ltd
Priority date: 2022-06-24
Filing date: 2023-06-21
Publication date: 2023-12-26

Abstract

The present disclosure relates to pyrimidinone derivatives and uses thereof, and in particular, provides pyrimidinone compounds represented by formula (I) or pharmaceutically acceptable salts thereof, which are useful for preparing medicines, particularly medicines for preventing and/or treating FXIa mediated diseases or conditions. The groups in the formula (I) are defined in the specification.

Description

Pyrimidinone derivative and pharmaceutical composition, preparation method and application thereof

The present disclosure claims priority from a prior application filed 24-6-2022 to the intellectual property office of China, having patent application number 202210738748.6, entitled "pyrimidinone derivatives and pharmaceutical compositions, methods of preparation and uses thereof". The entirety of this prior application is incorporated by reference into this disclosure.

Technical Field

The present disclosure belongs to the field of medicine, and in particular relates to pyrimidinone derivatives serving as FXIa inhibitors, and a pharmaceutical composition, a preparation method and application thereof.

Background

Blood clotting is the result of the coordinated activation of various plasma proteins, cofactors and platelets. This cascade is divided into the endogenous (contact activation) pathway, the exogenous (tissue factor activation) pathway, and the common (prothrombin and thrombin generation) pathway. The most important physiological process in blood coagulation is the activation of tissue factors. Tissue factor forms a complex with factor VIIa, catalyzes the activation of Factor X (FX), which in turn cleaves prothrombin to produce activated thrombin (FIIa). Activated thrombin (FIIa) acts as a central catalytic enzyme in the clotting process, catalyzing the cleavage of fibrinogen to fibrin, and acting as a clotting function. The exogenous approach has less enzyme quantity and quick effect. The intrinsic pathway is the body's intrinsic coagulation pathway, activating factor twelve (FXIIa), factor XIa (FXIa), factor IXa (FIXa) and factor VIIIa (FVIIIa) by a cascade of reactions, which in turn activates factor Xa (FXa) and downstream central thrombin (FIIa). Thrombin in turn activates factor XIa (FXIa), producing an amplifying effect that accelerates clotting. The intrinsic pathway is involved in more thrombin and is entirely from blood, generally with slower onset of action.

FXa plays a very critical role in the whole coagulation process. Antagonists thereof are widely used for the prevention and treatment of various thrombosis as downstream co-modulators of the extrinsic and intrinsic coagulation pathways. Various FXa antagonists are marketed, and occupy the cardiovascular medicine market due to their remarkable therapeutic effects. However, their occurrence probability of side effects is also relatively large, and most prominent is bleeding risk. To solve the bleeding problem, FXIa on the endogenous pathway is a research hotspot for various companies and institutions.

The potential of FXIa as a safer anticoagulation target is demonstrated in hemophilia C patients. FXIa-deficient hemophilia C patients have no active bleeding, which is evident in comparison to the ease of bleeding in factor VIIIa-deficient hemophilia A and factor IXa-deficient hemophilia B patients. While limited sample number (115 patients) studies indicate that FXIa deficiency does not protect patients from acute myocardial ischemia, such patients are found to have a lower incidence of ischemic stroke and deep vein thrombosis.

Knockout mice experiments have found that selective knockout of common pathway factors (FX, FV and FII) and exogenous factors (tissue factor and FVII) in mice results in prenatal or perinatal mortality. FVIII and FIX knockout mice, while viable, are often accompanied by severe bleeding, similar to haemophilia a and B, where deficiency of FVIII and FIX in humans poses a serious bleeding risk. Mice with selective FXI knockout can normally reproduce. Furthermore, FXI deficiency protects mice against ferric chloride-induced arterial thrombosis. Meanwhile, the absence of FXI does not affect the bleeding and hemostatic functions of the mice. Thus, this experiment shows that inhibition of FXI not only prevents thrombosis, but is also safely tolerated.

Many antibodies, small molecules and antisense nucleotides to FXIa have also been shown to be effective in preventing thrombosis in animals or clinically by inhibiting FXIa. But the risk of bleeding is greatly reduced compared to existing antithrombotic agents (e.g. enoxaparin). The above shows that FXIa is closely related to human thrombotic diseases, and inhibiting FXIa has remarkable anticoagulation effect, but has no obvious bleeding tendency, and can greatly reduce the bleeding risk in the clinical anticoagulation process. Therefore, the development of the medicine with good anticoagulation effect and small side effect has important research significance.

Patents for FXIa inhibitors which have been disclosed so far are WO2014154794, WO2014160592, WO2017005725, etc.

Disclosure of Invention

The present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1a is-C (O) R ⁶ Or a 5-membered heteroaryl, wherein said 5-membered heteroaryl is optionally substituted with one or more R ^A Substituted;

each R is ^A Identical or different and are each independently selected from halogen, C _1-6 Alkyl, C _1-6 Haloalkyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy, C _1-6 Hydroxyalkyl, cyano, carboxyl, 3 to 8 membered cycloalkyl and 3 to 8 membered heterocyclyl;

R ⁶ is H or C _1-6 An alkyl group;

R ^1b is H or halogen;

R ^1c And R is ^1d Identical or different and are each independently selected from H, halogen, C _1-6 Alkyl, C _1-6 Haloalkyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy, C _1-6 Hydroxyalkyl, cyano, amino, nitro, hydroxy and 3 to 8 membered cycloalkyl;

R ^2a is C _1-6 Alkoxy or halogen;

R ^2b selected from H, halogen, C _1-6 Alkyl, C _1-6 Haloalkyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy, C _1-6 Hydroxyalkyl, cyano and 3 to 8 membered cycloalkyl;

R ³ selected from C _1-6 Alkyl, C _1-6 Alkoxy, 3 to 8 membered cycloalkyl, 3 to 8 membered heterocyclyl, 6 to 10 membered aryl and 5 to 10 membered heteroaryl, wherein said C _1-6 Alkyl, C _1-6 Alkoxy, 3 to 8 membered cycloalkyl, 3 to 8 membered heterocyclyl, 6 to 10 membered aryl and 5 to 10 membered heteroaryl optionally being substituted by one or more groupsMultiple R' s ^3A Substitution;

each R is ^3A Identical or different and are each independently selected from halogen, C _1-6 Alkoxy, C _1-6 Haloalkoxy, hydroxy, cyano, amino, 3 to 8 membered cycloalkyl, 3 to 8 membered heterocyclyl, 3 to 8 membered cycloalkyloxy, 3 to 8 membered heterocyclyloxy, 6 to 10 membered aryl and 5 to 10 membered heteroaryl, wherein said 3 to 8 membered cycloalkyl, 3 to 8 membered heterocyclyl, 3 to 8 membered cycloalkyloxy, 3 to 8 membered heterocyclyloxy, 6 to 10 membered aryl and 5 to 10 membered heteroaryl are optionally selected from halogen, C _1-6 One or more substituents of alkyl, hydroxy and cyano are substituted;

ring a is a 6 to 10 membered aryl or a 5 to 10 membered heteroaryl;

each R is ⁴ Identical or different and are each independently selected from H, halogen, C _1-6 Alkyl, C _1-6 Haloalkyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy, C _1-6 Hydroxyalkyl, cyano, amino, nitro, hydroxy, 3 to 8 membered cycloalkyl and 3 to 8 membered heterocyclyl;

R ⁵ selected from H, C (O) OH and C (O) NH ₂ ；

n is selected from 0, 1, 2, 3 and 4.

In some embodiments, the present disclosure provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein R ^1a Selected from-C (O) R ⁶ 、R ⁶ Is H or C _1-6 An alkyl group; r is R ⁷ And R is ⁸ Identical or different and are each independently selected from H, halogen, C _1-6 Alkyl, C _1-6 Haloalkyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy, C _1-6 Hydroxyalkyl and cyano.

In some embodiments, the present disclosure provides compounds of formula (I), or a pharmaceutically acceptable salt thereof, wherein ring a is phenyl.

In some embodiments, the present disclosure provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereofWherein R is ^1a Selected from acetyl, propionyl,

in some embodiments, the present disclosure provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein R ^1c And R is ^1d Identical or different and are each independently selected from H, halogen, C _1-6 Alkyl and C _1-6 A haloalkyl group.

In some embodiments, the present disclosure provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein R ^1c And R is ^1d All are H.

In some embodiments, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a compound of formula (II) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ and n is as defined in formula (I).

In some embodiments, the present disclosure provides a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, which is a compound of formula (II-1) or formula (II-2) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ and n is as defined in formula (II).

In some embodiments, the present disclosure provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereofR in (B) ⁶ Is methyl or ethyl.

In some embodiments, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a compound of formula (III) or formula (IV) or a pharmaceutically acceptable salt thereof,

wherein:

R ⁷ and R is ⁸ Identical or different and are each independently selected from H, halogen, C _1-6 Alkyl, C _1-6 Haloalkyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy, C _1-6 Hydroxyalkyl and cyano;

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ and n is as defined in formula (I).

In some embodiments, the present disclosure provides a compound of formula (I) or formula (III) or a pharmaceutically acceptable salt thereof, which is a compound of formula (III-1) or formula (III-2) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ 、R ⁷ 、R ⁸ and n is as defined in formula (III).

In some embodiments, the present disclosure provides a compound of formula (I) or formula (IV) or a pharmaceutically acceptable salt thereof, which is a compound of formula (IV-1) or formula (IV-2) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ 、R ⁸ and n is as defined in formula (IV).

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ^2a Methoxy or F.

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ^2a Is methoxy.

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ^2a F.

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ^2b Selected from H, halogen and C _1-6 An alkyl group.

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ^2b H.

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ^1b H or F.

In some embodiments, the present disclosure provides a compound of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (I) V-2) or a pharmaceutically acceptable salt thereof, wherein R ⁵ is-COOH.

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ⁵ is-CONH ₂ 。

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ³ Is C _1-6 Alkyl, wherein said C _1-6 Alkyl is optionally substituted with one or more R ^3A Substitution;

R ^3A identical or different and are each independently selected from halogen, C _1-6 Alkoxy, C _1-6 Haloalkoxy, 3-to 8-membered cycloalkyl, 3-to 8-membered heterocyclyl and phenyl, said 3-to 8-membered cycloalkyl, 3-to 8-membered heterocyclyl and phenyl being optionally selected from halogen, C _1-6 One or more substituents in the alkyl and cyano groups.

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2), or pharmaceutically acceptable salts thereof, wherein R ³ Selected from methyl, ethyl, propyl, -CH ₂ CHF ₂ 、-CH ₂ CH ₂ OCH ₃ 、-CH ₂ CH ₂ OCF ₃ 、-CH ₂ CH ₂ OCHF ₂ 、

In some embodiments, the present disclosure provides compounds of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), and formula (IV-2)Or a pharmaceutically acceptable salt thereof, wherein R ³ Is C _1-6 Alkyl, wherein each R ⁴ Is H or halogen; n is selected from 0, 1 and 2.

Exemplary specific compounds of the present disclosure have structures including, but not limited to, the structures in table a below:

table A

Exemplary specific compounds of the present disclosure also include, but are not limited to, the structures in table B below:

table B

In another aspect of the present disclosure, there is provided an isotopic label for a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B.

In another aspect of the present disclosure, there is provided formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table AOr an isotopic label of a compound shown in table B, wherein said isotopic label is deuterium (D or ² H) Replaced hydrogen ¹ H)。

In another aspect of the present disclosure, there is provided a compound represented by formula (IA) or a salt thereof,

Wherein:

R ^w is C _1-6 An alkyl group;

ring A, R ^1a 、R ^1b 、R ^1c 、R ^1d 、R ^2a 、R ^2b 、R ³ 、R ⁴ And n is as defined in formula (I). In another aspect of the present disclosure, there is provided a compound represented by formula (IIA) or a salt thereof,

wherein:

R ^w is C _1-6 An alkyl group;

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁶ and n is as defined in formula (II).

In another aspect of the present disclosure, there is provided a compound represented by formula (IIIA) or a salt thereof,

wherein:

R ^w is C _1-6 An alkyl group;

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁷ 、R ⁸ and n is as defined in formula (III).

In another aspect of the present disclosure, there is provided a compound represented by formula (IVA) or a salt thereof,

wherein:

R ^w is C _1-6 An alkyl group;

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁸ and n is as defined in formula (IV).

In another aspect of the present disclosure, there is provided a compound represented by formula (IB) or a salt thereof,

wherein:

R ^1a 、R ^1b 、R ^1c 、R ^1d 、R ^2a 、R ^2b and R is ³ And n is as defined in formula (I).

In another aspect of the present disclosure, there is provided a compound represented by formula (IIB) or a salt thereof,

wherein: r is R ^1b 、R ^2a 、R ^2b 、R ³ And R is ⁶ As defined in formula (II).

In another aspect of the present disclosure, there is provided a compound represented by formula (IIIB) or a salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁷ and R is ⁸ As defined in formula (III).

In another aspect of the present disclosure, there is provided a compound represented by formula (IVB) or a salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ and R is ⁸ As defined in formula (IV).

In another aspect of the present disclosure, there is provided a method for preparing a compound represented by formula (I) or a pharmaceutically acceptable salt thereof, comprising the steps of,

Subjecting a compound represented by the formula (IA) or a salt thereof to hydrolysis reaction to obtain a compound represented by the formula (I) or a pharmaceutically acceptable salt thereof,

wherein:

R ^w is-C _1-6 An alkyl group; r is R ⁵ -COOH;

ring A, R ^1a 、R ^1b 、R ^1c 、R ^1d 、R ^2a 、R ^2b 、R ³ 、R ⁴ And n is as defined in formula (I).

Referring to the preparation method of the formula (I), the compound shown in the formula (IA) is replaced by a compound shown in the formula (IIA), a compound shown in the formula (IIIA) or a compound shown in the formula (IIVA), and the compound shown in the formula (II), the formula (III) or the formula (IV) can be obtained through hydrolysis reaction respectively; wherein R is ^w is-C _1-6 An alkyl group; r is R ⁵ is-COOH.

the compound shown in the formula (IB) or salt thereof and the compound shown in the formula (IC) or salt thereof undergo condensation reaction to obtain the compound shown in the formula (I) or pharmaceutically acceptable salt thereof,

wherein:

ring A, R ^1a 、R ^1b 、R ^1c 、R ^1d 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ And n is as defined in formula (I).

Referring to the preparation method of the compound shown in the formula (I) or the pharmaceutically acceptable salt thereof, the compound shown in the formula (IB) is replaced by a compound shown in the formula (IIB), a compound shown in the formula (IIIB) and a compound shown in the formula (IIVB), and a compound shown in the formula (IIC)The compound shown in the formula (II), the formula (III) or the formula (IIV) can be obtained by condensation reaction of the compound shown in the formula (III) or the salt thereof.

In another aspect of the present disclosure, a pharmaceutical composition is provided that includes at least one compound provided herein, or a pharmaceutically acceptable salt thereof, in a therapeutically effective amount, and one or more pharmaceutically acceptable excipients.

In another aspect of the present disclosure, there is also provided the use of a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, for the preparation of a medicament for inhibiting factor XIa.

The present disclosure also provides the use of a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table a or table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the foregoing, in the manufacture of a medicament for the prevention and/or treatment of a factor XIa-mediated disease or condition.

The present disclosure also provides the use of a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table a or table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, as described above, in the manufacture of a medicament for the prevention and/or treatment of a blood coagulation-related disease; preferably, the blood coagulation related disease is thrombosis or thromboembolic disease or cardiovascular and cerebrovascular disease.

The present disclosure also provides a method of inhibiting factor XIa comprising administering to a patient in need thereof a therapeutically effective amount of a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table a or table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

The present disclosure also provides a method of preventing and/or treating a factor XIa-mediated disease or condition comprising administering to a patient in need thereof a therapeutically effective amount of a compound as shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table a or table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

The present disclosure also provides a method for preventing and/or treating a blood coagulation-related disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table a or table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same; preferably, the blood coagulation related disease is thrombosis or thromboembolic disease or cardiovascular and cerebrovascular disease.

The present disclosure also provides a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the foregoing, for use as a medicament.

The present disclosure also provides a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the foregoing, for use as an FXIa inhibitor.

The present disclosure also provides a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, as a medicament for preventing and/or treating a factor XIa-mediated disease or disorder.

The present disclosure also provides a use of a compound shown in formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table a or table B, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same, as a medicament for preventing and/or treating a blood coagulation-related disease; preferably, the blood coagulation related disease is thrombosis or thromboembolic disease or cardiovascular and cerebrovascular disease.

Factor XIa mediated diseases or conditions described in the present disclosure are blood coagulation related diseases, preferably thrombosis or thromboembolic diseases or cardiovascular and cerebrovascular diseases.

The present disclosure "thrombotic or thromboembolic disorders" includes disorders that occur in the arterial and venous vascular system and that can be treated with the compounds of the present disclosure, particularly in the coronary arteries of the heart, such as Acute Coronary Syndrome (ACS), myocardial infarction with ST elevation (STEMI) and without ST elevation (non-STEMI), stable angina, unstable angina, reocclusion and restenosis following coronary interventions such as angioplasty, stent implantation or aortic coronary bypass surgery, and other vascular thrombotic or thromboembolic disorders that result in peripheral arterial occlusive disorders, pulmonary embolism, venous thromboembolism, venous thrombosis, particularly in the deep veins of the lower extremities, transient ischemic attacks, and thrombotic and thromboembolic strokes.

In some embodiments, the pharmaceutical composition is in a unit dose of 0.001mg to 1000mg.

In some embodiments, the pharmaceutical composition contains 0.01-99.99% of the aforementioned compound of formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B, or a pharmaceutically acceptable salt or isotopic label thereof, based on the total weight of the composition. In certain embodiments, the pharmaceutical compositions comprise 0.1-99.9% of a compound as set forth in the foregoing formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B, or a pharmaceutically acceptable salt or isotopic label thereof. In certain embodiments, the pharmaceutical compositions comprise 0.5% to 99.5% of a compound as set forth in the foregoing formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B, or a pharmaceutically acceptable salt or isotopic label thereof. In certain embodiments, the pharmaceutical compositions comprise 1% to 99% of a compound as set forth in the foregoing formula (I), formula (II-1), formula (II-2), formula (III-1), formula (III-2), formula (IV-1), formula (IV-2), table A or Table B, or a pharmaceutically acceptable salt or isotopic label thereof.

In certain embodiments, the pharmaceutical composition contains 0.01% to 99.99% of one or more pharmaceutically acceptable excipients, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1% to 99.9% of one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition contains 1% to 99% of one or more pharmaceutically acceptable excipients.

When used as a medicament, the compounds of the present disclosure may be administered in the form of a pharmaceutical composition. These compositions may be prepared in a manner well known in the pharmaceutical arts and may be administered by a variety of routes, depending upon whether local or systemic treatment and the area being treated is desired. Topical (e.g., transdermal, dermal, ocular, and mucosal including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal), oral, or parenteral administration. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration. Parenteral administration may be in the form of a single bolus dose or may be administered by, for example, a continuous infusion pump.

In preparing the compositions of the present disclosure, the active ingredient is typically admixed with excipients, which may be in the form of: tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or in liquid vehicles), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions and sterile packaged powders.

The "excipient" described in the present disclosure refers to ingredients other than the active ingredient, and includes, for example, diluents, fillers, absorbents, wetting agents, binders, disintegrants, lubricants, and the like.

On the other hand, pharmaceutically acceptable salts of the compounds described in this disclosure may be inorganic salts or organic salts, which may form acid addition salts if the compounds have a basic center; if these compounds have an acidic center, they may form base addition salts; these compounds may also form internal salts if they contain both acidic (e.g., carboxyl) and basic (e.g., amino) centers.

In another aspect, the compounds of the present disclosure may exist in particular geometric or stereoisomeric forms. For example, cis and trans isomers, (D) or (+) isomers ("(D)" or "(+)" means dextro), (L) or (-) isomers ("(L)" or "(-)" means levorotatory), (R) -and (S) -enantiomers, diastereomers, racemic and other mixtures, and enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the disclosure. 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 disclosure.

In the chemical structure of the compounds of the present disclosure, the bondIndicating unspecified configuration, ++>Or->Represents the absolute configuration of a stereocenter, i.e., the bond +.>Can be +.>Or->Or at the same time contain->And->Two configurations.

Key with a keyIndicating unspecified configurations including cis (E) or trans (Z) configurations.

In addition, the compounds and intermediates of the present disclosure may also exist in different tautomeric forms, and all such forms are included within the scope of the disclosure. "tautomer" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also known as proton transfer tautomers) include tautomers via proton transfer, such as keto-enol isomerisation, imine-enamine isomerisation and lactam-lactam isomerisation. All tautomeric forms of the compounds in the present disclosure are within the scope of the present disclosure. The names of compounds named in a singular manner do not exclude any tautomers. For exampleIf such is the case, it is understood that either individual forms of both tautomers or mixtures of both tautomers are within the scope of the disclosure.

The present disclosure also includes some isotopically-labeled compounds of the present disclosure having the same structure as recited herein, but wherein one or more atoms are replaced by an atom having an atomic weight or mass number different from the atomic weight or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of H, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as respectively ² H、 ³ H、 ¹¹ C、 ¹³ C、 ¹⁴ C、 ¹³ N、 ¹⁵ N、 ¹⁵ O、 ¹⁷ O、 ¹⁸ O、 ³¹ P、 ³² P、 ³⁵ S、 ¹⁸ F、 ¹²³ I、 ¹²⁵ I and ³⁶ cl, and the like. All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are intended to be encompassed within the scope of the present disclosure.

The term "therapeutically effective amount" of the present disclosure refers to that amount of active compound or drug substance that elicits the biological or medicinal response that is being sought in a tissue, system, animal, individual or human by a researcher, veterinarian, medical doctor or other clinician, which includes one or more of the following: (1) prevention of disease: for example, preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but has not experienced or developed a pathology or symptomatology of the disease. (2) inhibition of disease: for example, inhibiting a disease, disorder or condition (i.e., preventing further development of pathology and/or symptoms) in an individual experiencing or presenting with the pathology or symptoms of the disease, disorder or condition. (3) alleviation of disease: for example, alleviating a disease, disorder or condition (i.e., reversing the pathology and/or symptoms) in an individual experiencing or presenting with the pathology or symptoms of the disease, disorder or condition. For a drug or pharmacologically active agent, a "therapeutically effective amount" refers to a sufficient amount of the drug or agent that is non-toxic but achieves the desired effect. 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 present disclosure "pharmaceutically acceptable" refers 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 patients without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio, and are effective for the intended use.

The term "patient" in this disclosure refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses or primates, most preferably humans.

Description of the terms

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing from 1 to 20 carbon atoms, preferably an alkyl group containing from 1 to 12 (e.g., 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) carbon atoms, more preferably an alkyl group containing from 1 to 6 carbon atoms (C _1-6 Alkyl). Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2, 3-dimethylbutyl, and various branched isomers thereof, and the like. Alkyl groups may be substituted or unsubstituted.

The term "alkoxy" refers to-O- (alkyl), wherein alkyl is as defined herein. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy and butoxy. Alkoxy groups may be substituted or unsubstituted.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring comprising 3 to 20 carbon atoms, preferably comprising 3 to 12 carbon atoms or 3 to 8 (e.g. 3, 4, 5, 6, 7 and 8) carbon atoms, more preferably comprising 3 to 6 carbon atoms, said ring atoms optionally being oxo, the oxo (= O) group on the ring being part of a ring. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyl groups include spiro, fused and bridged cycloalkyl groups.

The term "spirocycloalkyl" refers to a 5 to 20 membered, polycyclic group sharing one carbon atom (referred to as a spiro atom) between each single ring in the system, which may contain one or more double bonds. Preferably 6 to 14 membered, more preferably 7 to 10 membered (e.g. 7, 8, 9 or 10 membered). The spirocycloalkyl group is classified into a single spirocycloalkyl group, a double spirocycloalkyl group or a multiple spirocycloalkyl group according to the number of common spiro atoms between rings, and preferably a single spirocycloalkyl group and a double spirocycloalkyl group. More preferably 3-membered/5-membered, 3-membered/6-membered, 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered, mono-spirocycloalkyl. Non-limiting examples of spirocycloalkyl groups include:

The term "fused ring alkyl" refers to 5 to 20 membered, all carbon polycyclic groups in which each ring in the system shares an adjacent pair of carbon atoms with the other rings in the system, wherein one or more of the rings may contain one or more double bonds. Preferably 6 to 14 membered, more preferably 7 to 10 membered (e.g. 7, 8, 9 or 10 membered). The number of constituent rings may be classified into a bicyclic, tricyclic, tetra-cyclic or polycyclic condensed ring alkyl group, preferably a bicyclic or tricyclic, more preferably a 3-membered/4-membered, 3-membered/5-membered, 3-membered/6-membered, 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/4-membered, 5-membered/5-membered, 5-membered/6-membered, 6-membered/3-membered, 6-membered/4-membered, 6-membered/5-membered and 6-membered/6-membered, and the like. Non-limiting examples of fused ring alkyl groups include:

the term "bridged cycloalkyl" refers to an all-carbon polycyclic group of 5 to 20 members, any two rings sharing two carbon atoms that are not directly attached, which may contain one or more double bonds. Preferably 6 to 14 membered, more preferably 7 to 10 membered (e.g. 7, 8, 9 or 10 membered). Cycloalkyl groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings are preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl groups include:

the cycloalkyl ring includes cycloalkyl (including monocyclic, spiro, fused, and bridged rings) fused to an aryl, heteroaryl, or heterocycloalkyl ring as described herein, wherein the ring attached to the parent structure is cycloalkyl, non-limiting examples include Etc.; preferably->The cycloalkyl group may be substituted or unsubstituted.

The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic substituent comprising 3 to 20 ring atoms, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, which may optionally be oxo (i.e. form a sulfoxide or sulfone), but excluding the ring portion of-O-, -O-S-or-S-, the remaining ring atoms being carbon, which ring carbon atoms may optionally be oxo, the oxo (=o) on the ring being part of a ring. Preferably from 3 to 12 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12) ring atoms, of which 1 to 4 (e.g., 1,2,3, and 4) are heteroatoms; more preferably 3 to 8 ring atoms (e.g., 3, 4, 5, 6, 7, and 8), of which 1-3 (e.g., 1,2, and 3) are heteroatoms; more preferably 3 to 6 ring atoms, of which 1-3 are heteroatoms; most preferably 5 or 6 ring atoms, of which 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, tetrahydropyranyl, 1,2,3, 6-tetrahydropyridinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclyl groups include spiro, fused and bridged heterocyclic groups.

The term "spiroheterocyclyl" refers to a 5 to 20 membered, polycyclic heterocyclic group in which each single ring in the system shares one atom (referred to as the spiro atom) and in which one or more of the ring atoms is a heteroatom selected from nitrogen, oxygen and sulfur, which sulfur may optionally be oxo (i.e. form a sulfoxide or sulfone), the remaining ring atoms being carbon. Which may contain one or more double bonds. Preferably 6 to 14 membered, more preferably 7 to 10 membered (e.g. 7, 8, 9 or 10 membered). The spiroheterocyclyl groups are classified into a single spiroheterocyclyl group, a double spiroheterocyclyl group or a multiple spiroheterocyclyl group according to the number of common spiro atoms between rings, and preferably a single spiroheterocyclyl group and a double spiroheterocyclyl group. More preferably 3-membered/5-membered, 3-membered/6-membered, 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered single spiro heterocyclyl. Non-limiting examples of spiroheterocyclyl groups include:

the term "fused heterocyclyl" refers to a 5 to 20 membered, polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with the other rings in the system, one or more of which may contain one or more double bonds, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, which may optionally be oxo (i.e., form sulfoxides or sulfones), and the remaining ring atoms are carbon. Preferably 6 to 14 membered, more preferably 7 to 10 membered (e.g. 7, 8, 9 or 10 membered). The number of constituent rings may be classified into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 3-membered/4-membered, 3-membered/5-membered, 3-membered/6-membered, 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/4-membered, 5-membered/5-membered, 5-membered/6-membered, 6-membered/3-membered, 6-membered/4-membered, 6-membered/5-membered and 6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclyl groups include:

The term "bridged heterocyclyl" refers to a 5 to 14 membered, polycyclic heterocyclic group in which any two rings share two atoms which are not directly connected, which may contain one or more double bonds, wherein one or more of the ring atoms is a heteroatom selected from nitrogen, oxygen and sulfur, which may optionally be oxo (i.e., form sulfoxides or sulfones), the remaining ring atoms being carbon. Preferably 6 to 14 membered, more preferably 7 to 10 membered (e.g. 7, 8, 9 or 10 membered). Heterocyclic groups which may be classified as bicyclic, tricyclic, tetracyclic or polycyclic bridged according to the number of constituent rings are preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclyl groups include:

such heterocyclyl rings include heterocyclyl (including monocyclic, spiro, fused and bridged heterocycles) fused to an aryl, heteroaryl or cycloalkyl ring as described herein, wherein the ring attached to the parent structure is heterocyclyl, non-limiting examples of which include:

etc. The heterocyclic group may be substituted or unsubstituted.

The term "aryl" refers to a 6 to 14 membered all-carbon monocyclic or fused polycyclic (fused polycyclic being a ring sharing adjacent pairs of carbon atoms) group having a conjugated pi-electron system, preferably 6 to 10 membered, such as phenyl and naphthyl. Such aryl groups include aryl rings fused to heteroaryl, heterocyclyl, or cycloalkyl rings as described herein, wherein the ring attached to the parent structure is an aryl ring, non-limiting examples of which include:

Aryl groups may be substituted or unsubstituted.

The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 (e.g., 1, 2, 3, and 4) heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaryl is preferably 5 to 10 membered (e.g., 5, 6, 7, 8, 9, or 10 membered), more preferably 5 or 6 membered, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, and the like. Such heteroaryl rings include heteroaryl fused to an aryl, heterocyclyl, or cycloalkyl ring as described herein, wherein the ring attached to the parent structure is a heteroaryl ring, non-limiting examples of which include:

heteroaryl groups may be substituted or unsubstituted.

The terms "alkyl", "alkoxy", "cycloalkyl", "heterocyclyl", "aryl" and "heteroaryl" and the like herein may be substituted or unsubstituted; when substituted, it may be substituted at any useful point of attachment, preferably the substituents are independently selected from the same or different substituents of one or more of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

The cycloalkyl, heterocyclyl, aryl and heteroaryl groups described above include residues derived from the removal of one H from the parent ring atom, or residues derived from the removal of two H from the same or two different ring atoms of the parent, i.e. "divalent cycloalkyl", "divalent heterocyclyl", "arylene", "heteroarylene".

The term "cycloalkyloxy" refers to a cycloalkyl-O-, wherein cycloalkyl is as defined herein.

The term "heterocyclyloxy" refers to heterocyclyl-O-, wherein heterocyclyl is as defined herein.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined herein.

The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein the alkoxy group is as defined herein.

The term "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxyl groups, wherein alkyl is as defined herein.

The term "halogen" refers to F, cl, br or I.

The term "hydroxy" refers to-OH.

The term "amino" refers to-NH ₂ 。

The term "cyano" refers to-CN.

The term "nitro" refers to-NO ₂ 。

The term "oxo" or "oxo" refers to "=o".

The term "carbonyl" refers to c=o.

The term "carboxy" refers to-C (O) OH.

The term "carboxylate" refers to-C (O) O (alkyl), -C (O) O (cycloalkyl), (alkyl) C (O) O-or (cycloalkyl) C (O) O-, wherein alkyl and cycloalkyl are as defined herein.

"optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "heterocycloalkyl group optionally substituted with an alkyl group" means that an alkyl group may be, but is not necessarily, present, and the description includes cases where the heterocycloalkyl group is substituted with an alkyl group and cases where the heterocycloalkyl group is not substituted with an alkyl group.

"substituted" means that one or more H in the group, preferably up to 5, more preferably 1 to 3 atoms, are independently substituted with a corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free H may be unstable when bound to carbon atoms having unsaturated (e.g., olefinic) bonds.

Advantageous effects of the present disclosure

The present disclosure provides a compound shown in formula (I), formula (II-1), formula (II-2), formula (IG), formula (IIG-1), formula (IIG-2), formula (IIIG-1), formula (IIIG-2), table A or Table B, which has a good FXIa inhibition effect, and can be used for effectively treating or preventing cardiovascular and cerebrovascular diseases and thromboembolic diseases. And compared with the disclosed FXIa inhibitor patent compound, the compound has excellent anticoagulation effect on human blood and good drug-substituted absorption activity.

Detailed Description

The technical scheme of the present disclosure will be described in further detail below with reference to specific embodiments. It should be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the present disclosure. All techniques implemented based on the foregoing disclosure are intended to be within the scope of the disclosure.

Unless otherwise indicated, the starting materials and reagents used in the following examples were either commercially available or may be prepared by known methods.

The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.

The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) or/and Mass Spectrometry (MS). NMR shift (. Delta.) of 10 ^-6 Units of (ppm) are given. NMR was performed using a Bruker Avance III-400 MHz nuclear magnetic resonance apparatus with deuterated dimethyl sulfoxide (DMSO-d) ₆ ) Deuterated chloroform (CDCl) ₃ ) Deuterated methanol (CD) ₃ OD), internal standard is Tetramethylsilane (TMS).

Mass Spectra (MS) were determined by Waters 2767HPLC/Waters SQD, waters H-class UPLC-SQD2, agilent HPLC/Waters liquid phase Mass Spectrometry.

Chiral HPLC analysis was performed using Shimadzu LC-20AD.

The thin layer chromatography silica gel plate is used for forming GF254 silica gel plate of chemical industry (Shanghai) limited company, the specification of the silica gel plate used by the Thin Layer Chromatography (TLC) is 0.2-0.25 mm, and the specification of the thin layer chromatography separation and purification product is 0.4-0.5 mm.

Column chromatography is generally used for 100-200 mesh silica gel of chemical industry (Shanghai) limited company as a carrier.

High performance liquid phase preparation used Waters HPLC, gilson HPLC and Biotage MPLC preparative chromatography.

Chiral separation column chromatography was performed using GilsonGX-281 preparative HPLC.

In the examples, unless otherwise specified, the reactions were carried out under nitrogen atmosphere.

The nitrogen atmosphere is defined as the reaction flask being connected to a nitrogen balloon of about 1 liter volume.

The hydrogen atmosphere is defined as the reaction flask being connected to a hydrogen balloon of about 1 liter volume.

In the examples, the reaction temperature is room temperature and the temperature range is 20-30 ℃ unless otherwise specified.

It will be appreciated by those skilled in the art that resolved chiral compounds can be distinguished by the order of retention times in the chiral chromatographic column, and therefore chiral compounds resolved sequentially for retention times are correspondingly distinguished by the numbered suffixes P1, P2. Namely, the suffix P1 corresponds to the chiral structure which is split first, and the suffix P2 corresponds to the chiral structure which is split later. The absolute configuration of the listed compounds in the chemical formula does not mean that the compounds with the numbers P1 and P2 are in one-to-one correspondence, but only the different existence forms of the absolute configuration are shown. The absolute configuration of the compounds numbered suffixes P1, P2 is subject to the absolute configuration that corresponds objectively to a particular retention time.

Detailed Description

Synthesis of intermediate Compound A1

Pinacol biborate (129.20 g,0.51 mmol), potassium acetate (83 g,0.85 mmol) and Pd (dppf) Cl at room temperature ₂ (7.38 g,0.01 mmol) was added to a solution of compound A1-1 (70 g,0.34 mmol) in DMSO (700 mL) and the reaction mixture was heated to 80℃and stirred at that temperature for 16 hours. After the completion of the reaction, the reaction mixture was diluted with water (1000 mL), extracted with ethyl acetate (1000 ml×2), the combined organic phases were washed with water (500 ml×3), concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=100/1 to 30/1) to give compound A1 (61 g, yield 75.2%). ¹ H NMR(400MHz,CDCl ₃ ):δ7.54(d,J＝2.8Hz,1H),7.14-7.11(m,1H),6.53(d,J＝8.8Hz,1H),4.70(brs,2H),1.33(s,12H)。

Synthesis of intermediate compound A2

The first step: synthesis of Compound A2-2

POCl at room temperature ₃ (9.20 g,0.030 mmol) was slowly added to a solution of compound A2-1 (3.12 g, 0.020mmol) in toluene (6 mL) and after heating the reaction mixture to 80℃and stirring at that temperature for one hour, triethylamine (4.04 g,0.040 mmol) was slowly added and stirring continued for 30 minutes. After the completion of the reaction, the reaction mixture was poured into 120mL of ice water, stirred for 12 hours, ph=5 was adjusted with aqueous sodium hydroxide solution, extracted with ethyl acetate (80 ml×3), the combined organic phases were washed with saturated brine (80 ml×1), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give compound A2-2 (2.0 g, yield 57.5%). ¹ H NMR(400MHz,CDCl ₃ ):δ8.12(s,1H),4.02(s,3H),3.94(s,3H)。MS m/z(ESI):174.9[M+1] ⁺ 。

And a second step of: synthesis of Compound A2-3

At room temperature, compound A1 (7 g,0.028 mmol) and Na were sequentially added ₂ CO ₃ (4.90 g,0.046 mmol) and Pd (dppf) Cl ₂ (0.84 g,1.15 mmol) DME/EtOH/H added to Compound A2-2 (4 g,0.023 mmol) ₂ O (100 mL/20mL/20 mL) and the reaction mixture was stirred at 90℃for 16 hours. After the completion of the reaction, the reaction mixture was diluted with water (500 mL), extracted with ethyl acetate (80 ml×3), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=5/1 to 2/1) to give compound A2-3 (4.70 g, yield 77.2%). ¹ H NMR(400MHz,CDCl ₃ ):δ8.29(s,1H),7.66(d,J＝2.4Hz,1H),7.16(d,J＝8.4Hz,1H),6.70(d,J＝8.8Hz,1H),3.99(s,3H),3.87(s,3H)。MS m/z(ESI):266.1[M+1] ⁺ 。

And a third step of: synthesis of Compound A2-4

An aqueous solution (10 mL) of sodium nitrite (1.47 g,0.021 mmol) was added to a hydrochloric acid solution (4M, 100 mL) of Compound A2-3 (4.70 g, 0.020mmol) at 0deg.C, and after stirring the reaction mixture for 30 minutes, an aqueous solution (10 mL) of sodium azide (1.70 g,0.027 mmol) was added and the reaction mixture was stirred at room temperature for 16 hours. After the reaction, the pH of the reaction mixture was adjusted with a saturated aqueous sodium bicarbonate solution>7 ethyl acetate extraction (100 ml×4), combining the organic phases and concentrating under reduced pressure, and purifying the residue by silica gel column chromatography (PE/ea=5/1 to 3/1) to give compound A2-4 (3.70 g, yield 71.7%). ¹ H NMR(400MHz,CDCl ₃ ):δ8.25(s,1H),7.45-7.39(m,2H),7.18(d,J＝8.4Hz,1H),3.99(s,3H),3.86(s,3H)。

Fourth step: synthesis of Compound A2-5

Trimethylsilylacetylene (3.40 g,0.012 mmol) and cuprous oxide (0.33 g,2.34 mmol) were added to a solution of compound A2-4 (3.40 g,0.012 mmol) in acetonitrile (100 mL) at room temperature, and the reaction mixture was heated to 90℃under nitrogen and stirred at that temperature for 16 hours. After the completion of the reaction, the reaction mixture was cooled and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=5/1 to 3/1) to give compound A2-5 (4.50 g, yield 99.1%). MS m/z (E) SI):390.2[M+1] ⁺ 。

Fifth step: synthesis of Compound A2-6

NCS (2.06 g,15.40 mmol) and silica gel (0.70 g) were added to a solution of Compound A2-5 (2.00 g,5.14 mmol) in acetonitrile (100 mL) at room temperature, the reaction mixture was heated to 55℃under nitrogen atmosphere and stirred at that temperature for 48 hours, after the completion of the reaction, the reaction mixture was cooled and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/EA=5/1 to 3/1) to give Compound A2-6 (1 g, yield 55.6%). MS m/z (ESI): 352.1[ M+1 ]] ⁺ 。

Sixth step: synthesis of Compound A2

LiCl (1.08 g,26 mmol) and TsOH (1.76 g,10 mmol) were added to a solution of compound A2-6 (1.0 g,2.85 mmol) in isopropanol (30 mL) at room temperature and the reaction mixture was stirred at 85℃for 16 h. After the reaction was completed, the reaction mixture was cooled and diluted with water (150 mL), extracted with ethyl acetate (100 ml×4), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by beating with ethyl acetate (20 mL) to give compound A2 (0.63 g, yield 65.6%). ¹ H NMR(400MHz,DMSO-d ₆ ):δ11.66(s,1H),8.81(s,1H),7.90-7.76(m,4H),3.38(s,3H)。MS m/z(ESI):338.0[M+1] ⁺ 。

Synthesis of intermediate Compound A3

The first step: synthesis of Compound A3-2

Compound A1 (2.74 g,0.01 mol), sodium carbonate (1.92 g,0.018 mol) and Pd (PPh) were successively added at room temperature ₃ ) ₄ (0.52 g,0.45 mmol) was added to a mixed solution of compound A3-1 (1.50 g,9.04 mmol) in ethylene glycol dimethyl ether/ethanol/water (80 mL/10mL/10 mL), and the reaction mixture was heated to 90℃under a nitrogen atmosphere and stirred at that temperature overnight. After the reaction, the reaction mixture was cooled and diluted with water (100 mL), extracted with ethyl acetate (100 ml×3), the combined organic phases were washed with saturated brine (100 ml×1), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=10/1 to 5/1) Compound A3-2 (2.10 g, yield: 90.5%) was obtained. MS m/z (ESI): 258.0[ M+1 ]] ⁺ 。

And a second step of: synthesis of Compound A3-3

Isoamyl nitrite (0.89 g,7.59 mmol) was added to a solution of compound A3-2 (1.30 g,5.06 mmol) in acetonitrile (50 mL) at 0deg.C, and the reaction mixture was stirred at 0deg.C for 30 min before adding TMSN ₃ (0.87 g,7.59 mmol) and the reaction mixture was slowly warmed to room temperature and stirred for 16 hours. After the reaction was received, the reaction mixture was diluted with water (50 mL), extracted with ethyl acetate (30 ml×3), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=10/1 to 5/1) to give compound A3-3 (1.25 g, yield: 87.4%). MS m/z (ESI) 284.0[ M+1 ]] ⁺ 。

And a third step of: synthesis of Compound A3-4

Compound A3-3 (1.68 g,5.94 mmol) was added to a solution of trimethylethynyl silicon (2.90 g,0.030 mol) in toluene (30 mL), and after the reaction mixture was stirred at 120℃overnight, the reaction mixture was cooled and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/EA=10/1 to 5/1) to give compound A3-4 (1.80 g, yield: 79.6%). MS m/z (ESI): 382.0[ M+1 ]] ⁺

Fourth step: synthesis of Compound A3-5

TsOH.H at room temperature ₂ O (0.18 g,0.94 mmol) was added to a solution of Compound A3-4 (1.80 g,4.72 mmol) and NCS (3.79 g,0.028 mol) in acetonitrile (80 mL) and the reaction mixture was stirred at 50℃for 16 h. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=10/1 to 5/1) to give compound A3-5 (0.96 g, yield: 59.3%). MS m/z (ESI): 344.1[ M+1 ] ] ⁺ 。

Fifth step: synthesis of Compound A3

Aqueous sodium hydroxide (4M, 15 mL) was added to a solution of compound A3-5 (0.91 g,2.65 mmol) in 1, 4-dioxane (15 mL), and the reaction mixture was stirred at 80℃for 16 hours. After the reaction was completed, the reaction mixture was diluted with water (50 mL), extracted with ethyl acetate (30 ml×3), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/meoh=50/1 to 15/1) to give compound A3 (0.77 g, yield: 89.3%). MS m/z (ESI): 326.0[M+1] ⁺

Synthesis of intermediate Compound A4

The first step: synthesis of Compound A4-1

LiCl (210 mg,5.09 mmol) and TsOH (1.76 g,2.04 mmol) were added to a solution of compound A2-3 (1.0 g,1.02 mmol) in isopropanol (5 mL) at room temperature and the reaction mixture stirred at 90℃for 20 h. After the completion of the reaction, the reaction mixture was cooled and diluted with water (50 mL), extracted with ethyl acetate (30 mL. Times.4), the organic phase was concentrated under reduced pressure, and the residue was purified by beating with ethyl acetate (20 mL) to give Compound A4-1 (220 mg, yield: 84.6%). MS m/z (ESI): 252.1[ M+1 ]] ⁺ 。

And a second step of: synthesis of Compound A4

Trimethyl orthoformate (10 mL) and sodium azide (1.14 g, 0.020mol) were added sequentially to a solution of compound A4-1 (640 mg,2.55 mmol) in acetic acid (20 mL), and the reaction mixture was heated to 80℃under nitrogen atmosphere and stirred at that temperature for 16 hours. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: xbridge-C18; 19X 150mm,5 μm; mobile phase: acetonitrile-water (0.1% ammonium bicarbonate), gradient: 10-40%, column temperature: 25 ℃ C.; flow rate: 15mL/min; wavelength: 214nm; column pressure: 80 bar) to give Compound A4 (490 mg, yield: 54.7%). MS m/z (ESI) 305.1[ M+1 ] ] ⁺ 。

Synthesis of intermediate Compound A5

The first step: synthesis of Compound A5-2

Aqueous hydrobromic acid (40%, 8 mL) was added to a solution of Compound A5-1 (2 g,15.50 mmol) in water (40 mL) at 0deg.C, and after stirring the reaction mixture at 0deg.C for 10 minutes, a solution of sodium nitrite (1.30 g,18.60 mmol) in water (40 mL) was slowly added dropwise, and the reaction mixture was stirred at 0deg.C for 30 minutes and at 5deg.C for 16 hours. After completion of the reaction, ethyl acetate (100 mL. Times.3) was added to the reaction solution to extract, and the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give compound A5-2 (3 g, yield 100%).

And a second step of: synthesis of Compound A5

Parmethoxybenzyl chloride (3.50 g,21.99 mmol) was slowly added to a solution of compound A5-2 (2.10 g,10.99 mmol), potassium carbonate (4.55 g,32.97 mmol) and lithium bromide (2.87 g,32.97 mmol) in DMF (50 mL) at 0deg.C. The reaction mixture was slowly heated to 35 ℃ and stirred at that temperature for 16 hours. After the reaction was completed. The reaction solution was diluted with water (400 mL), extracted with ethyl acetate (200 ml×3), and the combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=40/1 to 10/1) to give compound A5 (1.10 g, yield 32.0%). ¹ H NMR(400MHz,CDCl3):7.53(d,J＝11.6Hz,2H),7.93(d,J＝11.6Hz,2H),5.19(s,2H),4.44-4.30(m,1H),3.86(s,3H),2.03-1.84(m,2H),0.88-0.75(m,1H),0.58-0.43(m,2H),0.21-0.02(m,2H)。

Synthesis of intermediate compound A6

Perchloric acid (0.75 mL) was added to a solution of compound A6-1 (15 g,0.066 mol) in t-butyl acetate (225 mL) at 0deg.C, and the reaction mixture was stirred at room temperature for 16 hours. After the completion of the reaction, saturated aqueous sodium hydrogencarbonate (500 mL) was added to the reaction solution, extracted with ethyl acetate (500 ml×3), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=100/1 to 20/1) to give a colorless oily product A6 (14.92 g, yield 80.1%). ¹ H NMR(400MHz,CDCl ₃ ):δ7.39-7.24(m,5H),4.38-4.31(m,1H),3.51-3.41(m,1H),3.30-3.20(m,1H),1.45(s,9H)。

Example 1 Synthesis of Compound 2

The first step: synthesis of Compound 2a

Methyl 2-bromobutyrate (331 mg,1.48 mmol) and potassium carbonate (204 mg,1.48 mmol) were successively added to a mixed solution of compound A2 (250 mg,0.74 mmol) in DMF and THF (8 mL/16 mL), and the reaction mixture was stirred at 60℃for 16 hours. After the reaction was completed, the reaction solution was cooled and diluted with water (10 mL), extracted with ethyl acetate (15 ml×3), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=5/1 to 1/1) to give compound 2a (200 mg, yield 56.2%). ¹ H NMR(400MHz,CDCl ₃ ):δ7.75(d,J＝2.0Hz,1H),7.64-7.62(m,2H),7.54(d,J＝8.8Hz,1H),7.19(s,1H),5.35-5.31(m,1H),3.37(s,3H),2.23-2.16(m,1H),1.93-1.85(m,1H),1.51(s,9H),1.00(t,J＝7.2Hz,3H)。

And a second step of: synthesis of Compound 2b

TFA (8 mL) was added to a solution of compound 2a (180 mg,0.37 mmol) in DCM (8 mL) at 0deg.C, and the reaction mixture was stirred at room temperature for 5 hours, after the completion of the reaction, the reaction mixture was concentrated under reduced pressure to give compound 2b (156 mg, yield 100%). MS m/z (ESI) 424.0[ M+1 ] ] ⁺ 。

And a third step of: synthesis of Compound 2c

Tert-butyl 4-aminobenzoate (85 mg,0.44 mmol), T were added sequentially at 0deg.C ₃ P (470.64 mg,0.74 mmol) and triethylamine (112 mg,1.11 mmol) were added to a solution of compound 2b (156 mg,0.37 mmol) in dichloromethane (10 mL) and the reaction mixture was stirred at room temperature for 16 h. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=10/1 to 3/1) to give compound 2c (150 mg, yield 67.8%). MS m/z (ESI): 599.1.1[ M+1 ]] ⁺ 。

Fourth step: synthesis of Compounds 2-P1 and 2-P2

TFA (8 mL) was added to a solution of compound 2c (150 mg,0.25 mmol) in DCM (8 mL) at 0deg.C and the reaction mixture was stirred at room temperature for 4 h. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: xbridge-C18; 19X 150mm,5 μm; mobile phase: acetonitrile-water (0.1% ammonium bicarbonate), gradient: 20-70%, column temperature: 25 ℃ C.; flow rate: 15mL/min; wavelength: 214nm; column pressure: 80 bar) to give Compound 2 (100 mg, yield 74.1%). Compound 2 (80 mg) was purified by chiral preparative chromatography [ apparatus: gilsonGX-281, column: chiralpark ID 250mm by 30mm 10 μm; mobile phase: hex/EtOH/tfa=40/60/0.2; flow rate: resolution of 25mL/min gave compound 2-P1 (31.3 mg, yield 39.1%) and compound 2-P2 (29.9 mg, yield 37.4%).

Compound 2-P1:

MS m/z(ESI):542.9[M+1] ⁺ chiral HPLC with retention time 4.52min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):δ10.79(s,1H),8.89(s,1H),7.93-7.87(m,3H),7.83-7.81(m,3H),7.73(d,J＝8.8Hz,2H),5.38(t,J＝8.0Hz,1H),3.38(s,3H),2.18-2.14(m,2H),0.90-0.85(m,3H)。

Compound 2-P2:

MS m/z(ESI):542.9[M+1] ⁺ chiral HPLC: retention time 6.80min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):δ10.79(s,1H),8.89(s,1H),7.93-7.87(m,3H),7.83-7.81(m,3H),7.73(d,J＝8.8Hz,2H),5.38(t,J＝8.0Hz,1H),3.38(s,3H),2.18-2.14(m,2H),0.90-0.85(m,3H)。

Example 2 Synthesis of Compound 4

The first step: synthesis of Compound 4a

Compound A5 (447 mg,1.42 mmol) was added to a mixed solution of compound A2 (240 mg,0.71 mmol) and potassium carbonate (196 mg,1.42 mmol) in DMF and THF (3 mL/6 mL) and the reaction mixture was stirred at 60℃for 16 h. After the reaction was completed, the reaction solution was cooled and diluted with water (10 mL), extracted with ethyl acetate (15 ml×3), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=5/1 to 1/1) to give compound 4a (160 mg, yield 39.5%). ¹ H NMR(400MHz,CDCl3):7.75(d,J＝2.4Hz,1H),7.65-7.62(m,2H),7.53(d,J＝8.8Hz,1H),7.30-7.26(m,2H),7.15(s,1H),6.90-6.87(m,2H),5.54-5.50(m,1H),5.15(s,2H),3.81(s,3H),3.30(s,3H),2.06-1.89(m,2H),0.65-0.60(m,1H),0.53-0.46(m,2H),0.11-0.02(m,2H)。

And a second step of: synthesis of Compound 4b

Concentrated hydrochloric acid (3 mL) was added to a solution of compound 4a (150 mg,0.26 mmol) in acetonitrile (6 mL), and the reaction mixture was stirred at room temperature for 16 hours. After the completion of the reaction, the reaction mixture was diluted with water (10 mL), extracted with ethyl acetate (15 ml×3), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=5/1 to 1/1) to give compound 4b (110 mg, yield 93.2%). MS m/z (ESI) 450.1[ M+1 ]] ⁺ 。

And a third step of: synthesis of Compound 4c

Tert-butyl 4-aminobenzoate (56 mg,0.29 mmol) and T were added sequentially at 0deg.C ₃ P (284 mg,0.45 mmol) and triethylamine (68 mg,0.67 mmol) were added to a solution of compound 4b (100 mg,0.22 mmol) in dichloromethane (5 mL) and the reaction mixture was stirred at room temperature for 16 h. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=10/1 to 1/1) to give compound 4c (110 mg, yield 79.1%). MS m/z (ESI) 625.0[ M+1 ]] ⁺ 。

Fourth step: synthesis of Compounds 4-P1 and 4-P2

TFA (4 mL) was added to a solution of compound 4c (110 mg,0.18 mmol) in DCM (4 mL) at 0deg.C and the reaction mixture was stirred at room temperature for 2 hours. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: xbridge-C18; 19X 150mm,5 μm; mobile phase: acetonitrile-water (0.1% ammonium bicarbonate), gradient: 10-45%, column temperature: 25 ℃ C.; flow rate: 15mL/min; wavelength: 214nm; column pressure: 80 bar) to give Compound 4 (70 mg, yield 70.0%). Compound 4 (70 mg) was purified by chiral preparative chromatography [ apparatus: gilsonGX-281, column: chiralpark ID 250mm by 30mm 10um; mobile phase: hex/EtOH/tfa=40/60/0.2; flow rate: resolution of 25mL/min gave compound 4-P1 (26.10 mg, 37.3%) and compound 4-P2 (24.20 mg, 34.6%).

Compound 4-P1:

MS m/z(ESI):569.0[M+1] ⁺ . Chiral HPLC: retention time 4.68min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):10.75(s,1H),8.87(s,1H),7.93-7.87(m,4H),7.82-7.80(m,2H),7.73(d,J＝8.8Hz,2H),5.58-5.54(m,1H),3.38(s,3H),2.27-2.18(m,1H),1.93-1.86(m,1H),0.62-0.58(m,1H),0.48-0.36(m,2H),0.18-0.06(m,2H)。

Compound 4-P2:

MS m/z(ESI):569.0[M+1] ⁺ . Chiral HPLC: retention time 7.28min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):10.75(s,1H),8.87(s,1H),7.93-7.87(m,4H),7.82-7.80(m,2H),7.73(d,J＝8.8Hz,2H),5.58-5.54(m,1H),3.38(s,3H),2.27-2.19(m,1H),1.93-1.87(m,1H),0.63-0.58(m,1H),0.50-0.36(m,2H),0.18-0.08(m,2H)。

Example 3 Synthesis of Compound 13

The first step: synthesis of Compound 13a

Methyl 2-bromo-3-phenylpropionate (0.72 g,2.97 mmol) and potassium carbonate (0.41 g,2.97 mmol) were added to a mixed solution of compound A2 (0.50 g,1.48 mmol) in DMF and THF (10 mL/20 mL), and the reaction mixture was stirred at 60℃for 16 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/CH ₃ Oh=100/1 to 50/1) to give compound 13a (0.18 g, yield 24.3%). MS m/z (ESI) 500.5[ M+1 ]] ⁺ 。

And a second step of: synthesis of Compound 13b

LiOH.H ₂ O (0.21 g,4.92 mmol) was added to Compound 13a (0.73 g,1.64 mmol) in THF and H ₂ O (20 mL/5 mL) and the reaction mixture was stirred at room temperature for 16 hours. After the reaction, the pH of the reaction solution is adjusted by dilute hydrochloric acid<7, extracted with ethyl acetate (50 mL. Times.3), and the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give compound 13b (0.20 g, yield 100%). MS m/z (ESI): 486.0[ M+1 ]] ⁺ 。

And a third step of: synthesis of Compound 13c

Sequentially adding 4-aminobenzene Methyl formate (0.088 g,0.45 mmol), T ₃ P (0.52 g,0.82 mmol) and triethylamine (0.13 g,1.24 mmol) were added to a solution of compound 13b (0.20 g,0.41 mmol) in dichloromethane (20 mL) and the reaction mixture was stirred at room temperature for 16 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/CH ₃ Oh=100/1 to 50/1) to give compound 13c (0.23 g, yield 76.9%). MS m/z (ESI): 661.1[ M+1 ]] ⁺ 。

Fourth step: synthesis of Compounds 13-P1 and 13-P2

TFA (2 mL) was added to a solution of compound 13c (0.20 g,0.30 mmol) in dichloromethane (8 mL) and the reaction mixture was stirred at room temperature for 16 h. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: xbridge-C18; 19X 150mm,5 μm; mobile phase: acetonitrile-water (0.1% ammonium bicarbonate), gradient: 15-50%, column temperature: 25 ℃ C.; flow rate: 15mL/min; wavelength: 214nm; column pressure: 80 bar) to give Compound 13 (0.16 g, yield 79.6%). Compound 13 (0.16 g) was purified by chiral preparative chromatography [ apparatus: gilsonGX-281, column: chiralpark ID 250mm by 30mm 10 μm; mobile phase: hex/EtOH/tfa=50/50/0.2; flow rate: resolution of 25mL/min gave compound 13-P1 (50 mg, 31.2%) and compound 13-P2 (54 mg, 33.8%).

Compound 13-P1:

MS m/z(ESI):605.2[M+1] ⁺ . Chiral HPLC: retention time 5.84min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):10.85(s,1H),8.84(s,1H),7.94(d,J＝8.8Hz,3H),7.88-7.84(m,1H),7.78(d,J＝8.4Hz,1H),7.73-7.69(m,3H),7.32(t,J＝7.2Hz,2H),7.21(d,J＝6.0Hz,3H),5.81-5.76(m,1H),3.57-3.42(m,2H),3.33(s,3H)。

Compound 13-P2:

MS m/z(ESI):605.2[M+1] ⁺ chiral HPLC: retention time 9.48min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):10.84(s,1H),8.84(s,1H),7.92(t,J＝8.8Hz,3H),7.87-7.84(m,1H),7.78(d,J＝8.8Hz,1H),7.72-7.69(m,3H),7.32(t,J＝8.0Hz,2H),7.21(d,J＝6.0Hz,3H),5.81-5.77(m,1H),3.57-3.42(m,2H),3.33(s,3H)。

Example 4 Synthesis of Compound 29

The first step: synthesis of Compound 29a

Potassium carbonate (314 mg,2.28 mmol) was added to a mixed solution of compound A3 (370 mg,1.14 mmol) and compound A6 (389 mg,1.37 mmol) in THF and DMF (20 mL/10 mL), and the reaction mixture was stirred at 65℃for 16 hours. After the completion of the reaction, the reaction mixture was diluted with water (50 mL), extracted with ethyl acetate (30 ml×3), the organic phases were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=20/1 to 2/1) to give compound 29a (100 mg, yield: 16.6%). MS m/z (ESI) 530.0[ M+1 ]] ⁺ 。

And a second step of: synthesis of Compound 29b

TFA (3 mL) was added to a solution of compound 29a (100 mg,0.19 mmol) in DCM (9 mL) at 0deg.C, and the reaction mixture was stirred at room temperature for 2 hours, after the completion of the reaction, the reaction solution was concentrated under reduced pressure to give compound 29b (90 mg, crude). MS m/z (ESI) 474.1[ M+1 ]] ⁺ 。

And a third step of: synthesis of Compound 29c

T is carried out at 0 DEG C ₃ P (288 mg,0.45 mmol) and triethylamine (69 mg,0.68 mmol) were added to a solution of compound 29b (107 mg,0.23 mmol) and tert-butyl 4-aminobenzoate (48 mg,0.25 mmol) in dichloromethane (5 mL), and the reaction mixture was stirred at room temperature for 16 h. After the completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE/ea=3/1 to 2/1) to give compound 29c (110 mg, yield: 74.8%). MS m/z (ESI): 649.0[ M+1 ] ] ⁺ 。

Fourth step: synthesis of Compounds 29-P1 and 29-P2

TFA (3 mL) was added to a solution of compound 29c (110 mg,0.17 mmol) in DCM (9 mL) at 0deg.C and the reaction mixture was stirred at room temperature for 2 h. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: xb ridge-C18; 19X 150mm,5 μm; mobile phase: acetonitrile-water (0.1% TFA); gradient: 30-70%; column temperature: 25 ℃ C.; flow rate: 15mL/min; wavelength: 214nm; column pressure: 80 bar) to give compound 29 (95 mg, yield 95.0%).

Compound 29 (95 mg) was purified by chiral preparative chromatography [ apparatus: gilsonGX-281, column: chiralpark ID 250mm by 30mm 10um; mobile phase: hex/EtOH/tfa=50/50/0.2; flow rate: resolution of 25mL/min gave compound 29-P1 (20.1 mg, 21.0%) and compound 29-P2 (17.3 mg, 18.2%).

Compound 29-P1:

MS m/z(ESI):593.0[M+1] ⁺ . Chiral HPLC: retention time 5.43min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d6):δ12.71(br s,1H),10.93(s,1H),8.93(s,1H),8.81(d,J＝6.0Hz,1H),7.96-7.93(m,3H),7.86-7.84(m,2H),7.72(d,J＝8.8Hz,2H),7.35-7.31(m,2H),7.25-7.22(m,3H),5.81-5.76(m,1H),3.50-3.44(m,2H)。

Compound 29-P2:

MS m/z(ESI):593.0[M+1] ⁺ . Chiral HPLC: retention time 8.74min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d6)):δ12.71(br s,1H),10.93(s,1H),8.96(s,1H),8.81(d,J＝6.0Hz,1H),7.96-7.93(m,3H),7.86-7.84(m,2H),7.72(d,J＝8.8Hz,2H),7.35-7.31(m,2H),7.25-7.22(m,3H),5.81-5.76(m,1H),3.50-3.37(m,2H)。

Example 5 Synthesis of Compound 36

The first step: synthesis of Compound 36a

Compound A5 (733 mg,2.35 mmol) and potassium carbonate (324 mg,2.35 mmol) were added sequentially to a mixed solution of compound A4 (356 mg,1.17 mmol) in DMF and THF (20 mL/40 mL), and the reaction mixture was stirred at 60℃for 16 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/CH ₃ Oh=100/1 to 50/1) to give compound 36a (290 mg, yield: 47.0%). MS m/z (ESI): 537.1[ M+1 ]] ⁺ 。

And a second step of: synthesis of Compound 36b

Concentrated hydrochloric acid (6 mL) was added to a solution of compound 36a (270 mg,0.50 mmol) in acetonitrile (12 mL)In solution, the reaction mixture was stirred at room temperature for 16 hours. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by a C18 reverse column (mobile phase: acetonitrile-water (0.1% ammonium bicarbonate); gradient: 5-95%) to give compound 36b (160 mg, yield 76.2%). MS m/z (ESI) 417.1[ M+1 ]] ⁺ 。

And a third step of: synthesis of Compound 36c

Will T ₃ P (4819 mg,0.77 mmol) and triethylamine (117 mg,1.15 mmol) were added to a solution of compound 36b (160 mg,0.38 mmol) and tert-butyl 4-aminobenzoate (82 mg,0.42 mmol) in dichloromethane (10 mL), and the reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/CH ₃ Oh=50/1 to 20/1) to give compound 36c (97 mg, yield: 42.9%). MS m/z (ESI): 592.1[ M+1 ]] ⁺ 。

Fourth step: synthesis of Compound 36-P1 and Compound 36-P2

TFA (3 mL) was added to a solution of compound 36c (100 mg,0.17 mmol) in dichloromethane (9 mL) and the reaction mixture was stirred at room temperature for 2 hours. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: xbridge-C18; 19X 150mm,5 μm; mobile phase: acetonitrile-water (0.1% ammonium bicarbonate), gradient: 5-50%, column temperature: 25 ℃ C.; flow rate: 15mL/min; wavelength: 214nm; column pressure: 80 bar) to give compound 36 (90 mg, yield: 100%). Compound 36 (90 mg) was prepared by chiral [ apparatus: gilsonGX-281, column: chiralpark IE 250mm x 30mm 10um; mobile phase: hex/EtOH/tfa=40/60/0.2; flow rate: resolution of 25mL/min gave compound 36-P1 (43.6 mg, yield 48.4%) and compound 36-P2 (42.6 mg, yield 47.3%).

Compound 36-P1:

MS m/z(ESI):536.1[M+1] ⁺ . Chiral HPLC: retention time 6.76min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):12.74(br s,1H),10.75(s,1H),9.88(s,1H),7.94-7.85(m,6H),7.72(d,J＝8.8Hz,2H),5.56(q,J＝4.4Hz,1H),3.45(s,3H),2.24-2.16(m,1H),1.95-1.87(m,1H),0.63-0.58(m,1H),0.48-0.35(m,2H),0.16-0.06(m,2H)。

Compound 36-P2:

MS m/z(ESI):536.1[M+1] ⁺ . Chiral HPLC: retention time 10.63min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):12.75(br s,1H),10.77(s,1H),9.90(s,1H),7.94-7.88(m,6H),7.72(d,J＝8.8Hz,2H),5.56(q,J＝4.4Hz,1H),3.36(s,3H),2.24-2.16(m,1H),1.95-1.87(m,1H),0.63-0.58(m,1H),0.48-0.35(m,2H),0.16-0.06(m,2H)。

Example 6 Synthesis of Compound 43

The first step: synthesis of Compound 43a

Methyl 2-bromo-3-phenylpropionate (780 mg,3.20 mmol) and potassium carbonate (445 mg,3.22 mmol) were successively added to a mixed solution of compound A4 (490 mg,1.48 mmol) in DMF and THF (20 mL/40 mL), and the reaction mixture was stirred at 60℃for 16 hours. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM/CH ₃ Oh=100/1 to 50/1) to give compound 43a (178 mg, yield: 23.7%). MS m/z (ESI): 467.2[ M+1 ]] ⁺ 。

And a second step of: synthesis of Compound 43b

LiOH.H ₂ O (27 mg,0.64 mmol) was added to Compound 43a (150 mg,0.32 mmol) in THF and H ₂ O (20 mL/5 mL) and the reaction mixture was stirred at room temperature for 1 hour. After the reaction, the pH of the reaction solution is adjusted by dilute hydrochloric acid<7, extraction with ethyl acetate (30 mL. Times.3), washing the combined organic phases with saturated brine, drying over anhydrous sodium sulfate and filtration, and concentration of the filtrate under reduced pressure gave compound 43b (145 mg, yield: 100%). MS m/z (ESI): 453.0[ M+1 ]] ⁺ 。

And a third step of: synthesis of Compound 43c

Will T ₃ P (428 mg,0.66 mmol) and triethylamine (100 mg,0.99 mmol) were added to a solution of compound 43b (150 mg,0.33 mmol) and tert-butyl 4-aminobenzoate (71 mg,0.37 mmol) in dichloromethane (20 mL), and the reaction mixture was stirred at room temperature for 16 hr. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (DCM +.CH ₃ Oh=50/1 to 20/1) to give compound 43c (190 mg, yield: 91.3%). MS m/z (ESI): 628.3[ M+1 ]] ⁺ 。

Fourth step: synthesis of Compounds 43-P1 and 43-P2

TFA (2 mL) was added to a solution of compound 43c (190 mg,0.30 mmol) in dichloromethane (8 mL) and the reaction mixture was stirred at room temperature for 16 h. After the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: xbridge-C18; 19X 150mm,5 μm; mobile phase: acetonitrile-water (0.1% ammonium bicarbonate), gradient: 15-50%, column temperature: 25 ℃ C.; flow rate: 15mL/min; wavelength: 214nm; column pressure: 80 bar) to give Compound 43 (140 mg, yield: 80.9%). MS m/z (ESI) 572.1[ M+H ]] ⁺ . Compound 43 (140 mg) was purified by chiral preparative chromatography [ apparatus: gilsonGX-281, column: chiralpark ID 250mm by 30mm 10um; mobile phase: hex/EtOH/tfa=50/50/0.2; flow rate: 25mL/min ]Resolution yielded compound 43-P1 (51 mg, 36.4%) and compound 43-P2 (47 mg, 33.6%).

Compound 43-P1:

MS m/z(ESI):572.2[M+1] ⁺ . Chiral HPLC: retention time 6.33min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):10.84(s,1H),9.80(s,1H),7.95-7.85(m,5H),7.76-7.70(m,3H),7.33(t,J＝7.2Hz,2H),7.23(t,J＝8.4Hz,3H),5.81-5.76(m,1H),3.57-3.40(m,2H),3.31(s,3H)。

Compound 43-P2:

MS m/z(ESI):572.2[M+1] ⁺ . Chiral HPLC: retention time 9.88min, uv=254 nm. ¹ H NMR(400MHz,DMSO-d ₆ ):10.84(s,1H),9.80(s,1H),7.95-7.85(m,5H),7.76-7.70(m,3H),7.33(t,J＝7.2Hz,2H),7.23(t,J＝8.4Hz,3H),5.81-5.76(m,1H),3.57-3.42(m,2H),3.31(s,3H)。

Biological evaluation

Test example 1: determination of FXIa enzymatic Activity in vitro

The purpose of the experiment is as follows:

detection of inhibitory Activity of Compounds of the present disclosure against human FXIa factor

Experimental protocol:

the initial concentration of test compound was 10 μm, diluted 5-fold, 10 concentrations, each concentration was double-well. After the initial concentration solution of the intermediate dilution of each compound is mixed for 1min by shaking at 1000 rpm, 8 mu L of each compound is respectively added into 32 mu L of 100% DMSO to dilute 5 times, and the mixture is mixed for 1min by shaking at 1000 rpm. In this way, the compounds are diluted in a 5-fold gradient. 10 intermediate dilutions of the compound were prepared separately for each compound. The initial concentration, the multiple dilution ratio, the gradient concentration number and the compound hole number of the compound can be changed according to the actual condition of compound screening. The final concentration of Human Factor XIa (Haematologic Technologies Inc., HCXIA-0160) in the reaction system was 0.08nM and the final concentration of substrate D-LPR-ANSNH-C3H7 (Haematologic Technologies Inc., SN-13A) was 75. Mu.M, and after pre-incubating the reaction system in 384 well plates at 25℃for 10min, 4. Mu.L of 2.5 Xsubstrate D-LPR-ANSNH-C3H7 working solution was added to each well. Wherein, the positive control well contains enzyme, substrate, 0.5% DMSO and buffer solution, and no compound; the negative control wells contained substrate, 0.5% dmso and buffer, without enzyme and compound; the compound wells contained enzyme, substrate, compound, 0.5% dmso and buffer. 384 well plates (PerkinElmer, 6007270) were centrifuged at 1000 rpm for 30 seconds, gently mixed, and the reaction was started. Excitation light was set to 352nm and emission light was set to 470nm on a microplate reader, and kinetic readings were taken for 1 hour at 25 ℃. Raw data at 20min were collected for data processing and analysis, concentration-effect curves were fitted with GraphPad Prism 7 software, and compound concentration IC50 for 50% inhibition was calculated. Firstly, calculating the percentage inhibition rate corresponding to each compound concentration, after calculating the inhibition rate of each compound concentration, fitting a concentration-effect curve by using a 'log (inhibitor) vs. response-Variable slope (four parameters)' equation of GraphPad Prism 7 software, thereby obtaining the IC50. The correlation calculation formula is as follows: average value: calculation was performed using the AVERAGE formula of Excel. Standard deviation: calculation was performed using the STDEV formula of Excel. Z factor = 1- (3 x standard deviation of positive control well fluorescence intensity readings +3 x standard deviation of negative control well fluorescence intensity readings)/(mean of positive control well fluorescence intensity-mean of negative control table well fluorescence intensity), S/B = mean of positive control well fluorescence intensity/mean of negative control well fluorescence intensity, inhibition ratio (%) = (mean of positive control well fluorescence intensity-mean of compound well fluorescence intensity)/(mean of positive control well fluorescence intensity-mean of negative control well fluorescence intensity) ×100. The results are shown in Table 1 below:

TABLE 1

Compounds of formula (I)	FXIa IC ₅₀ (nM)
		Compound 4-P2	2.5
Compound 13-P2	0.13
		Compound 29-P2	0.72
Compound 36-P2	1.19
		Compound 43-P2	0.09

Conclusion: the compound disclosed by the disclosure has good inhibition effect on human factor XIa enzyme.

Test example 2: determination of aPTT (activated partial thrombin time), PT (prothrombin time) in vitro in rabbit plasma

The purpose of the experiment is as follows: detection of anticoagulant Effect of Compounds of the present disclosure on Rabbit plasma in vitro

aPTT (activated partial thrombin time) protocol:

in vitro aPTT experiment of rabbit plasma was performed using an activated partial thrombin time measurement kit (MDC, cat: 300025), a two-channel hemagglutination analyzer (German Meican MC-2000). Test compounds were dissolved in DMSO to 10mM stock solution and stored at-20deg.C for use. The mother liquor was thawed at the time of use, and the final concentrations of the test compounds were 60. Mu.M, 12. Mu.M, 2.4. Mu.M, 0.48. Mu.M, 0.096. Mu.M, 5 concentration gradients were used as negative controls using DMSO instead of the compound. The compound and plasma (venous whole blood collected in a non-fasted state, mixed with 3.2% sodium citrate anticoagulant at a ratio (v/v) of 9:1, 1560g x 8min, centrifuged at room temperature, and the supernatant extracted to prepare plasma) were mixed at a ratio of 1:49 to prepare a sample mixture. After the sample mixed solution is incubated for 3min at room temperature, the sample mixed solution is placed into a double-channel hemagglutination analyzer, incubated for 2min at 37 ℃, 30 mu L of aPTT reagent is added, after incubation is continued for 5min, 30 mu L of 0.025MCaCl2 is added, and the display value is read. Calculate the aPTT Ratio (aPTT Ratio = aPTT (sample)/aPTT (negative control)), map the existing data (working concentration, aPTT Ratio) using GraphPad Prism 7, calculate EC150 using OriginPro 2018SR1 version 9.5.1, EC200. Calculate the drug concentration at Ratio 1.5 and 2, i.e., EC150 and EC200, respectively, after fitting the Lg concentration and aPTT Ratio by exponential equation, results are shown in Table 2 below:

TABLE 2

Compounds of formula (I)	Rabbit plasma in vitro aPTT EC150 (mu M)
		Compound 13-P2	5.73

PT (prothrombin time) protocol:

the experiment was carried out using a prothrombin time measurement kit (MDC, cat:200353 PT), a two-channel hemagglutination analyzer (German Meicao brand MC-2000). Plasma treatment was carried out in accordance with the aPTT (activated partial thrombin time) experiment. After the test compound was mixed with plasma, the sample mixture was incubated at room temperature for 3min and at 37℃for 2min, 60. Mu.L of PT reagent preheated at 37℃was added vigorously, and the values were read. Calculation and statistical methods are performed with the aPTT (activated partial thrombin time) experiment.

Test example 3: determination of aPTT (activated partial thrombin time) in vitro in human plasma

The purpose of the experiment is as follows: detection of anticoagulant Effect of Compounds of the present disclosure on human plasma in vitro

Experimental protocol:

human plasma in vitro aPTT experiment was carried out using an activated partial thrombin time measurement kit (MDC, cat: 300025), a two-channel hemagglutination analyzer (German Meijuang brand MC-2000).

Test compounds were dissolved in DMSO to 10mM stock solution and stored at-20deg.C for use. The mother liquor was thawed at the time of use, and the final concentrations of the test compounds were 60. Mu.M, 12. Mu.M, 2.4. Mu.M, 0.48. Mu.M, 0.096. Mu.M, 5 concentration gradients were used as negative controls using DMSO instead of the compound. The compound and plasma (venous whole blood collected in a non-fasted state, mixed with 3.2% sodium citrate anticoagulant at a ratio (v/v) of 9:1, 1560g x 8min, centrifuged at room temperature, and the supernatant extracted to prepare plasma) were mixed at a ratio of 1:49 to prepare a sample mixture. After the sample mixed solution is incubated for 3min at room temperature, the sample mixed solution is placed into a double-channel hemagglutination analyzer, incubated for 2min at 37 ℃, 30 mu L of aPTT reagent is added, incubation is continued for 5min, then 30 mu L of 0.025M CaCl2 is added, and the display value is read. Calculate the aPTT Ratio (aPTT Ratio = aPTT (sample)/aPTT (negative control)), map the existing data using GraphPad Prism7 (working concentration/aPTT Ratio), calculate EC150 using originPro 2018SR1 version 9.5.1, EC200. Calculate the drug concentrations at Ratio 1.5, 2.0 and 3.0, i.e., EC150, EC200 and EC300, respectively, after fitting the Lg concentration and aPTT Ratio by exponential equation, the results are shown in Table 3 below:

TABLE 3 Table 3

Compounds of formula (I)	Human plasma in vitro aPTT EC150 (mu M)
		Compound 13-P2	1

Test example 4: determination of the Activity of the Exo Human Plasma Kalliliren enzyme

The purpose of the experiment is as follows:

detection of Human Plasma Kalliliren inhibitory Activity of Compounds of the present disclosure

Experimental protocol:

test compounds were dissolved in DMSO to 10mM stock solution and stored at-20deg.C for use. The initial concentration of the compound was 10. Mu.M, diluted 5-fold, 10 concentrations, 2. Mu.l/well, and final DMSO concentration was 0.5%; buffer solution preparation: 50mM tris-HCl, 150mM NaCl,0.01%Triton X-100, pH 7.4, 0.22. Mu.M filter sterilized; 2.5 Xhuman kallikrein protein (Natural human plasma kallikrein, abcam, cat#: ab77870, lot#: GR 251157-8): the final concentration of kallikrein was 1nM and the kallikrein was diluted to 2.5nM with buffer, 4. Mu.L/well. Preparing a 2.5 x substrate reaction solution: the final concentration of substrate Z-FR-AMC (Ji Biochemical, 208708) was 40. Mu.M, and the substrate 10mM stock was diluted to 100. Mu.M, 4. Mu.L/well with buffer. To 384 plates, 4. Mu.L of 2.5 Xkallikrein reaction solution was added per well, and the negative control wells were replaced with buffer; sequentially adding diluted 5 Xcompound working solution into corresponding wells of 2 mu L/well, replacing negative and positive control wells with 2.5% DMSO, mixing, and pre-incubating at 37deg.C for 10min; finally, 4 mu L of 2.5 times substrate reaction liquid is added into each hole, and the mixture is vibrated and mixed for more than 30 seconds. Reacting at 37deg.C for 30min, exciting light 342nm, emitting light 440nm, measuring fluorescence intensity, prism curve analysis, calculating IC ₅₀ Values.

Test example 5: in vivo pharmacokinetic evaluation in rats

The purpose of the experiment is as follows:

detection of pharmacokinetic parameters of compounds of the present disclosure in rats

Experimental protocol:

the experiment uses the solvent: DMAC: solutol: PBS (V/V) =10%: 10%:80% (v/v/v). The preparation method comprises the following steps: accurately weighing the required compound, adding a certain volume of DMAC according to the proportion, vortex mixing to dissolve completely, sequentially adding Solutol and PBS according to the proportion, and mixing to obtain the final product. The vehicle used in the intravenous (iv) administration group and the oral (po) administration group in the experiment was the same vehicle. Intravenous dose was 0.5mpk and oral dose was 3mpk. Experimental blood collection time points: group IV: 0.083,0.25,0.5,1,2,4,7, 24h. PO group: 0.25,0.5,1,2,4,7, 24h at each time point, jugular vein collected whole blood 200uL, EDTA-K2 anticoagulated, immediately centrifuged at 4000rpm for 5min at 4℃and the supernatant was frozen in a-80℃refrigerator. Treatment of plasma samples: after precipitation with an internal standard containing CH3CN/MeOH (1:1, v/v) precipitant, centrifugation was carried out at 14000rpm for 5min, the supernatant was taken for analysis by LC-MS/MS (AB Triple Quard 5500) to obtain plasma concentrations, and parameter calculation was carried out by means of a non-compartmental model version Winnolin 8.1.

Conclusion: the compounds of the present disclosure have good pharmacokinetic properties in rats.

Test example 6: AV-SHUNT method for rabbits

Test purpose:

the inhibition of the compounds on arteriovenous thrombosis in vivo is detected.

Experimental protocol:

healthy male New Zealand rabbits, weighing 2.0-3.0kg, were not fasted and were not prohibited from water before the experiment. Sodium pentobarbital was prepared as a 1.5% (W/V) solution in physiological saline. Straight surgical scissors, ophthalmic forceps, micro vascular forceps, micro scissors, micro forceps, arterial clamps, and Shimadzu balance AUW220D for standby. And (5) starting molding after 20min of compound infusion, ending molding after 60min of compound infusion, and stopping liquid medicine infusion. The molding method is as follows, new Zealand white rabbits are retrieved from suppliers and adapted to random groupings after one day, and Mao Bei skin is shaved after anesthesia. The operation exposes the right external jugular vein, the total jugular vein, and the left carotid artery of the animal. Pre-buried cotton thread (10)cm) is connected with the static common artery and the jugular vein. The passage is opened when the liquid medicine is infused for 20min, and the thrombus is induced and timing is started. After induction for 40min, closing the passage, taking down the middle through pipe, and taking out the pre-buried cotton thread. The blood was removed by repeated 8 times of staining on qualitative filter paper (medium speed 18cm, hangzhou specialty paper Co., ltd.). The weighing paper is placed on a balance, the balance is peeled, cotton threads stained with blood water are placed on the weighing paper, and the wet weight of thrombus is weighed and recorded. After the thrombus is removed, the experimental animal is euthanized. The thrombus weight of each animal was recorded, plotted using software GraphPad Prism 7, statistically analyzed using one-way analysis of variance (ANOVA), and compared to the control group by multiple comparison analysis (Dunnett's test), origin pro 2018SR1 version 9.5.1 calculated compound ED ₅₀ 。

Test example 7: BT test

The purpose of the experiment is as follows: test compound for length of bleeding in laboratory rabbits

Test protocol: healthy male New Zealand rabbits, weight 2.0-3.0kg, no fasting and no water forbidden before experiment, sodium pentobarbital, physiological saline solution prepared into 1.5% (W/V) solution, 0.9% (W/V) NaCl solution, and a timer.

The method comprises the following steps: a 0.9% nacl solution was formulated and preheated to 37 ℃. The animals were anesthetized with 1.5% sodium pentobarbital solution (2 mL/kg) by intravenous injection at the ear margin, and the hind paw hairs were shaved for use. The nail was cut off (about 1/3 of the fingertip) in the macroscopic vascular portion, timing was started after confirming bleeding and the paw was immersed in physiological saline at 37 ℃. Bleeding was observed, stopping bleeding, i.e., stopping the timer, and recording the duration. The statistical analysis was performed using software GraphPad Prism 7 mapping, single factor analysis of variance (ANOVA) and compared to the control group by multiple comparison analysis (Dunnett's test).

The second method is as follows: the retainer secures the animal, shaves the hair on the ear, wipes the rabbit ears with normal saline, cuts a wound on the vein at the edge of the ear with a standard incision device, begins timing after bleeding and covers the wound with filter paper, and records the bleeding time period by observing the bleeding. The statistical analysis was performed using software GraphPad Prism 7 mapping, single factor analysis of variance (ANOVA) and compared to the control group by multiple comparison analysis (Dunnett's test).

The embodiments of the present disclosure have been described above. However, the present disclosure is not limited to the above embodiments. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present disclosure should be included in the protection scope of the present disclosure.

Claims

1. A compound shown as a formula (I) or pharmaceutically acceptable salt thereof,

wherein:

R ⁶ is H or C _1-6 An alkyl group;

R ^1b is H or halogen;

R ^2a is C _1-6 Alkoxy or halogen;

R ³ selected from C _1-6 Alkyl, C _1-6 Alkoxy, 3 to 8-membered cycloalkyl, 3-to 8-membered heterocyclyl, 6-to 10-membered aryl and 5-to 10-membered heteroaryl, wherein said C _1-6 Alkyl, C _1-6 Alkoxy, 3 to 8 membered cycloalkyl, 3 to 8 membered heterocyclyl, 6 to 10 membered aryl and 5 to 10 membered heteroaryl optionally substituted with one or more R ^3A Substitution;

ring a is a 6 to 10 membered aryl or a 5 to 10 membered heteroaryl;

R ⁵ selected from H, C (O) OH and C (O) NH ₂ ；

n is selected from 0, 1, 2, 3 and 4.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein ring a is phenyl.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R ^1c And R is ^1d Identical or different and are each independently selected from H, halogen, C _1-6 Alkyl and C _1-6 A haloalkyl group; preferably, R ^1c And R is ^1d All are H.

4. A compound according to any one of claim 1 to 3, which is a compound represented by the formula (II) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ and n is as defined in claim 1.

5. The compound according to claim 4, which is a compound represented by the formula (II-1) or the formula (II-2) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ 、R ⁶ and n is as defined in claim 4.

6. The compound according to any one of claims 1, 4 and 5, or a pharmaceutically acceptable salt thereof, wherein R ⁶ Is methyl or ethyl.

7. A compound according to any one of claim 1 to 3, which is a compound represented by the formula (III) or formula (IV) or a pharmaceutically acceptable salt thereof,

wherein:

R ⁷ and R is ⁸ Identical or different and are each independently selected from H, halogen Element, C _1-6 Alkyl, C _1-6 Haloalkyl, C _1-6 Alkoxy, C _1-6 Haloalkoxy, C _1-6 Hydroxyalkyl and cyano;

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ and n is as defined in claim 1.

8. The compound according to claim 7, which is a compound represented by the formula (III-1) or the formula (III-2) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ 、R ⁷ 、R ⁸ and n is as defined in claim 7.

9. The compound according to claim 7, which is a compound represented by the formula (IV-1) or the formula (IV-2) or a pharmaceutically acceptable salt thereof,

wherein:

R ^1b 、R ^2a 、R ^2b 、R ³ 、R ⁴ 、R ⁵ 、R ⁸ and n is as defined in claim 7.

10. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, wherein R ^2a Methoxy or F.

11. A compound according to any one of claims 1 to 10 or a pharmaceutical thereofSalts of the above acceptable, wherein R ^2b H.

12. The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein R ⁵ is-COOH.

13. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, wherein R ⁵ is-CONH ₂ 。

14. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein R ³ Is C _1-6 Alkyl, wherein said C _1-6 Alkyl is optionally substituted with one or more R ^3A Substitution;

15. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, wherein R ³ Selected from methyl, ethyl, propyl, -CH ₂ CHF ₂ 、-CH ₂ CH ₂ OCH ₃ 、-CH ₂ CH ₂ OCF ₃ 、-CH ₂ CH ₂ OCHF ₂ 、

16. The compound according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, wherein each R ⁴ Is H or halogen;n is selected from 0, 1 and 2.

17. The compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from any one of the following compounds,

18. the compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from any one of the following compounds,

19. a pharmaceutical composition comprising at least one therapeutically effective amount of a compound according to any one of claims 1-18, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

20. Use of a compound according to any one of claims 1-18, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 19, in the manufacture of a medicament for inhibiting factor XIa.

21. Use of a compound according to any one of claims 1-18, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 19, in the manufacture of a medicament for the prevention and/or treatment of factor XIa mediated diseases.

22. Use of a compound according to any one of claims 1-18, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 19, in the manufacture of a medicament for the prevention and/or treatment of a blood coagulation related disease; preferably, the blood coagulation related disease is thrombosis or thromboembolic disease or cardiovascular and cerebrovascular disease.