Inhibition of human integrin alpha 5 beta 1

Abstract

Disclosed are compounds of Formula (I) which are useful in methods of treating or preventing diseases or conditions responsive to the inhibition of human α5β1 integrin by administering a compound of Formula (I) to a patient in need thereof. Many diseases and conditions can be treated by administering a compound of Formula (I).

Description

[0001] Cross-reference to related applications This application claims priority and interest in U.S. Provisional Application No. 63 / 687,186, filed August 26, 2024, and U.S. Provisional Application No. 63 / 584,383, filed September 21, 2023; the contents of each of these applications are incorporated herein by reference in their entirety.

[0002] Field of the Invention This application discloses a method for treating diseases and conditions by inhibiting α5β1 integrin. Diseases and conditions that respond to inhibition of α5β1 integrin include, for example, hypertension, such as pulmonary arterial hypertension (PAH).

[0003] background Fibronectin (Fn) is an extracellular matrix protein that coordinates complex cell adhesion and signaling during tissue development, remodeling, and diseases such as hypertension and heart failure via cell surface integrin receptors (integrins that bind to fibronectin, such as a5b1). Heart failure (HF) is a debilitating disease in which abnormal cardiac function leads to low or inadequate perfusion of the body's tissues and organs. Hypertension causes a variety of detrimental effects with high morbidity and mortality, including heart failure. One form of hypertension is pulmonary hypertension (PAH). PAH is a rare but extremely destructive disease in which normally low pulmonary artery pressure becomes elevated due to vasoconstriction and remodeling of the pulmonary vessels. Vasoconstriction and vascular remodeling increase the workload on the right side of the heart, causing right-sided cardiac hypertrophy, fibrosis, and ultimately heart failure.

[0004] Integrins are a family of transmembrane glycoprotein receptors that mediate cell-cell and cell-matrix interactions. Integrins are heterodimers with two distinct chains (α and β subunits). Eighteen α subunits and eight β subunits have been described in mammals. The integrin superfamily of cell surface receptors is formed by a variety of structurally and functionally related surface glycoproteins, in which the various receptors exist as heterodimers of non-covalently linked α and β subunits. At least 18 distinct α subunits and 8 distinct β subunits have been identified in mammals, and they are known to form more than 24 distinct receptors. Each integrin interacts specifically with defined extracellular ligands (including extracellular matrix proteins such as fibronectin, vitronectin, collagen, and cell surface molecules such as VCAM, ICAM, and PECAM) via a linear adhesion motif.

[0005] Integrin α5β1 is composed of the subunit ITGA5 (integrin α5) and integrin β1. Multiple integrins bind to fibronectin. Integrin α5β1 is selective for fibronectin because its interaction requires the 9th and 10th type II repeat sequences of fibronectin (FNIII-9 and FNIII-10). α5β1 integrin expression is primarily found in the vascular system and connective tissue. Not only is expression significantly enhanced in tumor blood vessels, but it is also significantly enhanced in tumor cells themselves across various cancer types, including colon, breast, ovarian, lung, and brain tumors. It is further expressed to varying degrees in multiple cell types, including fibroblasts, hematopoietic cells, immune cells, smooth muscle cells, and epithelial cells. High expression of α5β1 integrin has also been observed in fibrotic tissues (such as pulmonary fibrosis).

[0006] Integrin α5βl (a5b1 or α5β1) is composed of α5 (a5 or α5) and βl (b1 or β1) subunits. The a5 subunit forms a specific dimer with the β1 subunit and is widely expressed in most tissues. Integrin a5b1 regulates cell adhesion almost entirely through its interaction with fibronectin (via a short arginine-glycine-aspartate (RGD) adhesion motif). However, endothelial cells and platelets can bind to fibrin via a5b1. The interaction between a5b1 and fibronectin plays an important role in physiological and pathological angiogenesis and vascular integrity. Although endothelial cells express a variety of integrins, a5b1 plays a crucial role in the survival of endothelial cells on a temporary in vitro matrix, inhibiting apoptosis, and promoting proliferation. A5b1 expression is upregulated in patients with tumor-associated vascular systems and pulmonary hypertension. Consistent with the key functional role of receptor-ligand pairs, a5b1 ligand fibronectin is also upregulated in tumor tissues during wound healing.

[0007] Current treatments include vasodilators targeting Ca2+ channels or endothelin receptors. Novel approaches are needed to treat pulmonary hypertension, PAH, heart failure, and related conditions. Due to the limited current treatments for pulmonary hypertension, there is significant concern and need for other or alternative therapies for the treatment, stabilization, prevention, and / or delay of pulmonary hypertension. Several approaches have been developed to obtain more effective and / or less toxic drugs for the treatment of pulmonary hypertension. However, these approaches still have serious side effects, and the resulting drugs typically exhibit short half-lives and low bioavailability.

[0008] There remains a medical need for effective and safe oral α5β1 integrin inhibitors as an important complement to treatment devices for α5β1 integrin-mediated conditions. In particular, there is still an unmet need for small molecules designed to inhibit integrin α5β1, including the need for such orally administered small molecules that avoid the need for regular therapeutic infusions and the complications associated with this form of drug administration. Summary of the Invention

[0009] This invention particularly provides methods and compositions for treating diseases associated with enhanced integrin α5β1 expression or activity. Many normal physiological and disease processes require contact between cells and other cells and / or the extracellular matrix. Cell-matrix and cell-cell adhesion are regulated by multiple protein families, including integrins, selectins, cadherins, and immunoglobulins, and promote a variety of normal cellular functions such as proliferation, migration, differentiation, or survival. Cell adhesion is also key to a range of pathologies, and therefore, pharmacological disruption of cell adhesion interactions can provide therapeutic intervention mechanisms. Integrin superfamily members, adhesion molecules, play important roles in acute and chronic disease states such as cancer, inflammatory diseases, stroke, and neurodegenerative diseases. Therefore, integrins represent a complex field of biology.

[0010] In one aspect, the invention is characterized by a compound of formula (I) or a pharmaceutically acceptable salt thereof: in It is a 6- to 12-membered aryl ring structure (e.g., phenyl) or a 3- to 19-membered heterocyclic ring structure (e.g., a 3- to 15-membered heterocyclic ring structure, such as a 6-membered heteroaryl), wherein Optional ground cover halogen, C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl substitution, wherein C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl, optionally surrounded by one or more halogens, C 1-4 Alkoxy or C 1-6 Alkyl substitution, the C 1-6 Alkyl groups may optionally be substituted with one or more halogens; a is 1, 2, 3, 4, 5, 6, 7, or 8; and R a C 1-6 alkyl.

[0011] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof: in: It has a 6 to 12-membered aryl ring structure (e.g., phenyl), wherein Optional ground cover halogen, C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl substitution, wherein C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl, optionally surrounded by one or more halogens, C 1-4 Alkoxy or C 1-6 Alkyl substitution, the C 1-6 Alkyl groups may optionally be substituted with one or more halogens; a is 1, 2, 3, 4, 5, 6, 7, or 8; and R a C 1-6 alkyl.

[0012] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof: in: It is a 3 to 19-membered heterocyclic ring structure (e.g., a 3 to 15-membered heterocyclic ring structure, such as a 6-membered heteroaryl), wherein Optional ground cover halogen, C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl substitution, wherein C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl, optionally surrounded by one or more halogens, C 1-4 Alkoxy or C 1-6 Alkyl substitution. The C 1-6 Alkyl groups may optionally be substituted with one or more halogens; a is 1, 2, 3, 4, 5, 6, 7, or 8; and R a C 1-6 alkyl.

[0013] In some embodiments, the compound of formula (I) is the compound of formula (IA) or a pharmaceutically acceptable salt thereof: .

[0014] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein It is phenyl or 6-membered heteroaryl.

[0015] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for in R1 is C 1-4 alkoxy group; and R2 is a C group optionally substituted with one or more halogens. 1-4 Alkyl; or When R1 and R2 together form fused 5- to 8-membered heterocyclic alkyl rings, fused 5- to 6-membered heteroaryl rings, or fused 7- to 15-membered spirocyclic heterocyclic alkyl ring systems... It is a 9- to 19-membered heterocyclic group, wherein the fused ring is optionally surrounded by halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl or 4- to 8-membered heterocyclic alkyl ring-substituted, wherein C 1-4 Alkyl, C 3-6 Cycloalkyl or 4- to 8-membered heterocyclic alkyl rings optionally separated by one or more halogens or C 1-4 Alkyl substitution, the C 1-4 The alkyl group is optionally substituted with one or more halogens; and R3 is a C group optionally substituted with one or more halogens. 1-4 Alkyl group. In some embodiments, R1 is methoxy; R2 is fluorine; and R3 is a C group optionally substituted with one or more fluorine molecules. 1-4 alkyl.

[0016] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for in R4 and R 14 Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 alkyl; X1 is CR 5a R 5b or NR 5c R 5a H or C optionally substituted with one or more halogens 1-4 Alkyl, and R 5b C 1-4 Alkyl or C 3-6 cycloalkyl, wherein the C 1-4 alkyl or the C 3-6 The cycloalkyl group is optionally substituted with one or more halogens; or R 5a and R 5bTogether they form spirocyclic 3 to 8-membered heterocyclic alkyl or spirocyclic C 3-6 Cycloalkyl, wherein the 3- to 8-membered heterocycloalkyl or the C 3-6 The cycloalkyl group is optionally substituted with the following: C14 groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 6-membered spirocyclic heterocyclic alkyl or C 3-6 Spirocyclic cycloalkyl, the 4- to 6-membered spirocyclic heterocyclic alkyl or C 3-6 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 Alkyl groups may optionally be substituted with one or more halogens; R 5c C 1-4 Alkyl or C 3-6 cycloalkyl, wherein the C 1-4 alkyl or the C 3-6 The cycloalkyl group may optionally be substituted with one or more halogens; X2 is CR 6a R 6b -CH2CR 6a R 6b C=O, O or NR 6c ; R 6a and R 6b Each independently represents H and C. 1-4 Alkyl, 5- to 6-heteroaryl, 6-aryl, C 3-7 Cycloalkyl or 4- to 7-membered heterocycloalkyl, wherein the C 1-4 The alkyl group, the 5- to 6-membered heteroaryl group, the 6-membered aryl group, the C3-8 cycloalkyl group, or the 4- to 7-membered heterocycloalkyl group is optionally oxidized by one or more halogens or C 1-4 Alkyl substitution; R 6c C that is optionally substituted with one or more halogens 1-4 alkyl X3 is a direct bond, CR 7a R 7b , O or NR 7c ; R 7a and R 7b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 alkyl; R 7c C that is optionally substituted with one or more halogens 1-4 alkyl; X4 is CR 8a R 8b or NR 8c R 8a For H and R8b C is a halogen or optionally substituted with one or more halogens. 1-4 alkyl; R 8a and R 8b Together they form spirocyclic 3 to 8-membered heterocyclic alkyl or C 3-6 Cycloalkyl, wherein the 3- to 8-membered heterocycloalkyl or the C 3-6 The cycloalkyl group is optionally substituted with the following: C14 groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 8-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl, the 4 to 8-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group is optionally substituted with one or more halogens; and R 8c For H, C 1-4 Alkyl, 4- to 8-membered heterocyclic alkyl or C 3-6 cycloalkyl, wherein the C 1-4 Alkyl, the 4- to 8-membered heterocyclic alkyl or the C 3-6 The cycloalkyl group may optionally be substituted with one or more halogens; The conditions are that X2 and X3 cannot both contain O or N heteroatoms, and X3 and X4 cannot both contain O or N heteroatoms.

[0017] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for R4 and R 14 Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 alkyl; X1 is CR 5a R 5b or NR 5c R 5a H or C optionally substituted with one or more halogens 1-4 Alkyl, and R 5b C 1-4 Alkyl or C 3-6 cycloalkyl, wherein the C 1-4 alkyl or the C 3-6 The cycloalkyl group is optionally substituted with one or more halogens; or R 5a and R 5b Together they form spirocyclic 3 to 8-membered heterocyclic alkyl or spirocyclic C 3-6Cycloalkyl, wherein the 3- to 8-membered heterocycloalkyl or the C 3-6 The cycloalkyl group is optionally substituted with the following: C14 groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 6-membered spirocyclic heterocyclic alkyl or C 3-6 Spirocyclic cycloalkyl, the 4- to 6-membered spirocyclic heterocyclic alkyl or C 3-6 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 Alkyl groups may optionally be substituted with one or more halogens; R 5c C 1-4 Alkyl or C 3-6 cycloalkyl, wherein the C 1-4 alkyl or the C 3-6 The cycloalkyl group may optionally be substituted with one or more halogens; X2 is CR 6a R 6b -CH2CR 6a R 6b C=O, O or NR 6c ; R 6a and R 6b Each independently represents H and C. 1-4 Alkyl, 5- to 6-heteroaryl, 6-aryl, C 3-7 Cycloalkyl or 4- to 7-membered heterocycloalkyl, wherein the C 1-4 alkyl, the 5- to 6-membered heteroaryl, the 6-membered aryl, the C 3-8 The cycloalkyl group or the 4- to 7-membered heterocycloalkyl group is optionally converted by one or more halogens or C. 1-4 Alkyl substitution ;R 6c C that is optionally substituted with one or more halogens 1-4 alkyl X3 is a direct bond, CR 7a R 7b , O or NR 7c ; R 7a and R 7b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 alkyl; R 7c C that is optionally substituted with one or more halogens 1-4 alkyl; X4 is CR 8a R 8b or NR 8c R 8a For H and R 8bC is a halogen or optionally substituted with one or more halogens. 1-4 alkyl; R 8a and R 8b Together they form spirocyclic 3 to 8-membered heterocyclic alkyl or C 3-6 Cycloalkyl, wherein the 3- to 8-membered heterocycloalkyl or the C 3-6 The cycloalkyl group is optionally substituted with the following: C14 groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 8-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl, the 4 to 8-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group is optionally substituted with one or more halogens; and R 8c For H, C 1-4 Alkyl, 4- to 8-membered heterocyclic alkyl or C 3-6 cycloalkyl, wherein the C 1-4 Alkyl, the 4- to 8-membered heterocyclic alkyl or the C 3-6 The cycloalkyl group may optionally be substituted with one or more halogens; The conditions are that X2 and X3 cannot both contain O or N heteroatoms, and X3 and X4 cannot both contain O or N heteroatoms.

[0018] In some embodiments, R1 is a methoxy group; R2 is fluorine; and R3 is a C group optionally substituted with one or more fluorine groups. 1-4 Alkyl group. In some embodiments, R 14 H is a; and X1 is a CR 5a R 5b And R 5a Let H be the number of 'R', and R be the number of 'R'. 5b It is methyl or cyclopropyl. In some embodiments, R4 is H, fluorine, or methyl; X1 is CR. 5a R 5b And R 5a Let H be the number of 'R', and R be the number of 'R'. 5b X2 is methyl; X3 is O and X4 is CR. 7a R 7b And R 7a and R 7b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 Alkyl; or X3 is O and X2 is CR 6a R 7b And R 6a and R 6b Each is independently H; and X4 is CR. 8a R 8b And R 8a and R8b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 Alkyl, or R 8a and R 8b Together they form 4 to 7-membered heterocyclic alkyl rings or C 3-7 Cycloalkyl rings, wherein 4- to 7-membered heterocyclic alkyl or C 3-7 The cycloalkyl group is optionally substituted with: halogen; C-shaped groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 7-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl, the 4- to 7-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

[0019] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for in R 10a and R 10b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 Alkyl; or R 10a and R 10b Together they form a spiral ring C 3-6 cycloalkyl or spirocyclic 6-membered heterocyclic alkyl; R 11a and R 11b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 alkyl; R 12a and R 12b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 alkyl; R 13a It is methyl, ethyl, or cyclopropyl; R 13b For H; and R 14 C is H, a halogen, or optionally substituted with one or more halogens. 1-4 alkyl.

[0020] In some implementation schemes, R 10a and R 10b Each is independently H or methyl; R 11a and R 11b Each is independently H, methyl, or ethyl; R12a and R 12b Each is independently H or methyl; R 13a It is methyl, ethyl, or cyclopropyl; R 13b For H; and R 14 It is H, fluorine, or methyl. In some embodiments, R4 is H or fluorine; R 13a It is methyl, ethyl, or cyclopropyl; and R 14 For H. In some implementations, X2 is NR. 6c And R 6c C 1-4 alkyl.

[0021] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for , where R 13a It is H or methyl, and R4 is H, F or methyl.

[0022] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for , or , where R4 is H, fluorine or methyl.

[0023] In some implementations, R 10a R 10b and R 11a Each is either H or methyl. In some embodiments, R... 11b It can be H, methyl, or ethyl.

[0024] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for , R4 is H, a halogen, or a methyl group optionally substituted with one or more halogens; and R 9a and R 9b Each is independently H, a halogen, or a methyl group optionally substituted with one or more halogens. In some embodiments, R4 is H, fluorine, or methyl; and R 9a and R 9b Each of the following is independently H, fluorine, chlorine, or a methyl group optionally substituted with one or more fluorine molecules.

[0025] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for ,and in R 20 H, halogen, C 1-4 Alkyl or C 1-4 alkoxy group, wherein the C 1-4 Alkyl and C 1-4 Each alkoxy group may optionally be replaced by one or more halogens; X5 is a CR 25a R 25b or NR 25c ; R 25a and R 25b Each is independently H or optionally substituted with one or more halogens or alkoxy groups. 1-4 Alkyl; or R 25a and R 25b Together they form C 3-6 Cycloalkyl rings or 4- to 6-membered heterocyclic rings, each optionally coated with halogen, C 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or spirocyclic 4- to 6-membered heterocyclic cyclic substitution, wherein C 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or the spirocyclic 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens or C 1-4 Alkyl substitution; R 25c C 1-4 alkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 The alkyl group, the 5- to 6-membered heteroaryl group, or the C6 aryl group may optionally be affected by one or more halogens or C6 halogens. 1-4 Alkyl substitution; X6 is CR 26a R 26b Or C=O; R 26a and R 26b Each is independently H or optionally by one or more halogens or C 1-4 Alkyl-substituted C 1-4 alkyl; X7 is a CR 27a R 27b or NR 27c ; R 27a and R 27b Each is independently H or optionally substituted with one or more halogens or alkoxy groups. 1-4 Alkyl; or R 27a and R 27b Together they form C 3-6 Cycloalkyl ring or 4- to 6-membered heterocyclic ring, wherein the C3-6 The cycloalkyl ring or the 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens, one or more C-membered rings. 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or spirocyclic 4- to 6-membered heterocyclic cyclic substitution, wherein the C 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or the spirocyclic 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens or C 1-4 Alkyl substitution; and R 27c For H, C 1-4 Alkyl, C 3-6 Cycloalkyl, 4- to 7-membered heterocycloalkyl or 5- to 11-membered spirocycloheteroalkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 Alkyl, the C 3-6 Cycloalkyl, the 4- to 6-membered heterocycloalkyl, the 5- to 11-membered spirocycloheterocycloalkyl, the 5- to 6-membered heteroaryl, or the C6 aryl, optionally with one or more halogens or C 1-4 Alkyl substitution, the C 1-4 Alkyl groups may optionally be substituted with one or more halogens; The condition is that only one of X5 and X7 contains N heteroatoms.

[0026] In some implementation schemes, R 20 H, halogen, C 1-4 Alkyl or C 1-4 alkoxy group, wherein the C 1-4 alkyl and the C 1-4 The alkoxy group may optionally be substituted with one or more halogens; X5 is CR. 25a R 25b ;R 25a and R 25b Each is independently a methyl group; or R 25a and R 25b Together they form a cyclopropyl group; X6 is C=O; R 26a and R 26b Each is independently H or optionally by one or more halogens or C 1-4 Alkyl-substituted C 1-4 Alkyl; X7 is NR 27c And R 27c C 1-4 Alkyl, C 3-6 Cycloalkyl, 4- to 7-membered heterocycloalkyl or 5- to 11-membered spirocycloheteroalkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 Alkyl, the C 3-6 Cycloalkyl, the 4- to 6-membered heterocycloalkyl, the 5- to 11-membered spirocycloheterocycloalkyl, the 5- to 6-membered heteroaryl, or the C6 aryl, optionally with one or more halogens or C1-4 Alkyl substitution, the C 1-4 The alkyl group is optionally substituted with one or more halogens. In some embodiments, R 27c C 1-4 Alkyl group. In some embodiments, R 27c It is a 5-membered heteroaryl group. In some embodiments, R 27c C 3-6 Cycloalkyl groups, wherein the C atoms are optionally substituted with one or more halogens or optionally substituted with one or more halogens. 1-4 Alkyl substitution. In some embodiments, R 27c C 1-4 Alkyl, 4- to 7-membered heterocyclic alkyl or 5- to 11-membered spirocyclic heterocyclic alkyl, wherein the C 1-4 Alkyl groups, the 4- to 7-membered heterocyclic alkyl groups, and the 5- to 11-membered spirocyclic heterocyclic alkyl groups are optionally oxidized by one or more halogens or C. 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

[0027] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for or , Where R 20 It can be H, fluorine, methyl, ethyl, methoxy, -CH2-O-CH3, or -CF3.

[0028] In some implementation schemes, R 27c It is a tetrahydropyran optionally substituted with one or more methyl groups or fluorine groups. In some embodiments, R 27c for or , where R 32a R 32b R 33a and R 33b Each is independently H, methyl, or fluorine; or R 32a and R 32b Or R 33a and R 33b One of them together forms a spiral ring C 3-6 Cycloalkyl or spirocyclic 3- to 6-membered heterocyclic rings. In some embodiments, R 27c for .

[0029] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for in R 30a R 30b R 31a and R 31b Each can be independently H, fluorine, or methyl; R 20 It can be H, fluorine, methyl, ethyl, methoxy, -CH2-O-CH3, or -CF3; and R 27c The methyl group is present. In some embodiments, the compound is a compound of formula (I), wherein R is a methyl group. 27c C 1-4 alkyl.

[0030] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for .

[0031] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for Where R 20 H, halogen, optionally C 1-4 alkoxy-substituted C 1-4 alkoxy groups, C groups optionally substituted with 4 to 6-membered heterocyclic groups or one or more halogens 1-4 Alkyl group. In some embodiments, R 25c C 1-4 Alkyl; and R 27c To be optionally subjected to one or more halogens or C 1-4 alkoxy-substituted C 1-4 Alkyl, C 3-6 Cycloalkyl or 4- to 6-membered heterocycloalkyl, wherein the C 3-6 The cycloalkyl group or the 4- to 6-membered heterocycloalkyl group is optionally substituted with one or more halogens. 1-4 Alkyl substitution. In some embodiments, R 27c C 1-4 Alkyl; and R 25c To be optionally subjected to one or more halogens or C 1-4 alkoxy-substituted C 1-4 Alkyl, C 3-6 Cycloalkyl or 4- to 6-membered heterocycloalkyl, wherein the C 3-6 The cycloalkyl group or the 4- to 6-membered heterocycloalkyl group is optionally substituted with one or more halogens. 1-4 Alkyl substitution. In some embodiments, R20 It is H or methyl.

[0032] In some embodiments, the compound of formula (I), such as the compound of formula (IA) or a pharmaceutically acceptable salt thereof, wherein for or ;in R 20 H, halogen, optionally C 1-4 alkoxy-substituted C 1-4 alkoxy groups, C groups optionally substituted with 4 to 6-membered heterocyclic groups or one or more halogens 1-4 alkyl; R 25c C 1-4 Alkyl; and R 27c It is a 4- to 6-membered heterocyclic group or C 3-6 cycloalkyl, wherein the 4- to 6-membered heterocyclic group or the C 3-6 cycloalkyl groups are optionally C 1-4 Alkyl substitution.

[0033] In some embodiments, the compound of formula (I) is selected from the following: Or its pharmaceutically acceptable salt. Brief description of the attached diagram Figure 1 This is the list of compounds disclosed in this article.

[0035] Detailed description Compound (I) is a small molecule integrin therapeutic agent that targets α5β1 and can be administered to treat patients with diseases and symptoms that respond to inhibition of α5β1 integrin.

[0036] This invention provides methods and compositions for treating diseases associated with enhanced integrin α5β1 expression or activity, comprising administering an integrin α5β1 inhibitor. In some embodiments, the disease is characterized by World Health Organization (WHO) groups. In some embodiments, the disease is pulmonary hypertension, WHO Group 1 pulmonary hypertension or pulmonary arterial hypertension (PAH), WHO Group 2 pulmonary hypertension, WHO Group 3 pulmonary hypertension, WHO Group 4 pulmonary hypertension, and WHO Group 5 pulmonary hypertension. In some embodiments, the disease is characterized by the World Health Organization (WHO) classification system. In some embodiments, the disease is characterized by the WHO functional classification based on cardiac function. In some embodiments, the disease is pulmonary hypertension, WHO Group I pulmonary hypertension or pulmonary arterial hypertension (PAH), WHO Group II pulmonary hypertension, WHO Group III pulmonary hypertension, and WHO Group IV pulmonary hypertension. In some embodiments, the disease associated with enhanced integrin α5β1 expression or activity is heart failure or right ventricular failure.

[0037] Inhibiting integrin α5β1 by blocking its activity or inhibiting its binding to fibronectin can effectively prevent and treat pulmonary hypertension, PAH, heart failure, and right ventricular failure. This invention provides a variety of compounds (e.g., small molecule compounds and antibodies) that inhibit this interaction. These compounds are generally referred to herein as "integrin α5β1 inhibitors".

[0038] Exemplary compounds of the present invention This disclosure relates to novel compounds and methods for inhibiting α5β1 integrin.

[0039] Illustrative formulas and compounds are described herein. Illustrative embodiments of structural features that may exist in any of the formulas described herein are also provided herein. Any illustrative embodiment of a structural feature may be combined with any other illustrative structural feature described herein. Furthermore, and unless otherwise indicated herein, any description of a formula or compound also includes any pharmaceutically acceptable form of that compound, including but not limited to any pharmaceutically acceptable salts, hydrates, solvates, isomers, polymorphs, prodrugs, and isotopically labeled derivatives of the disclosed formulas and compounds.

[0040] Compound of formula (I) In some embodiments, the integrin α5β1 inhibitor is a small molecule compound that binds to integrin α5β1. In some embodiments, the integrin α5β1 inhibitor is a small molecule compound that specifically binds to integrin α5. In some embodiments, the integrin α5β1 inhibitor is a small molecule compound that specifically binds to integrin β1. In some embodiments, the integrin α5β1 inhibitor is a small molecule compound that specifically binds to integrin α5β1. In some embodiments, the integrin α5β1 inhibitor is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0041] In one aspect, the invention is characterized by a compound of formula (I) or a pharmaceutically acceptable salt thereof: in It is a 6- to 12-membered aryl ring structure (e.g., phenyl) or a 3- to 19-membered heterocyclic ring structure (e.g., a 3- to 15-membered heterocyclic ring structure, such as a 6-membered heteroaryl), wherein Optional ground cover halogen, C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl substitution, wherein C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl, optionally surrounded by one or more halogens, C 1-4 Alkoxy or C 1-6 Alkyl substitution, the C 1-6 Alkyl groups may optionally be substituted with one or more halogens; a is 1, 2, 3, 4, 5, 6, 7, or 8; and R a C 1-6 alkyl.

[0042] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof: in: It has a 6 to 12-membered aryl ring structure (e.g., phenyl), wherein Optional ground cover halogen, C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl substitution, wherein C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl, optionally surrounded by one or more halogens, C 1-4 Alkoxy or C1-6 Alkyl substitution, the C 1-6 Alkyl groups may optionally be substituted with one or more halogens; a is 1, 2, 3, 4, 5, 6, 7, or 8; and R a C 1-6 alkyl.

[0043] In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof: in: It is a 3 to 19-membered heterocyclic ring structure (e.g., a 3 to 15-membered heterocyclic ring structure, such as a 6-membered heteroaryl), wherein Optional ground cover halogen, C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl substitution, wherein C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl, optionally surrounded by one or more halogens, C 1-4 Alkoxy or C 1-6 Alkyl substitution, the C 1-6 Alkyl groups may optionally be substituted with one or more halogens; a is 1, 2, 3, 4, 5, 6, 7, or 8; and R a C 1-6 alkyl.

[0044] In some implementation schemes, It has a 3 to 15-membered heterocyclic structure.

[0045] In some implementations, the compound is a compound of formula (IA) or a pharmaceutically acceptable salt thereof.

[0046] In some embodiments, the compound is a compound of formula (I), wherein It is phenyl or 6-membered heteroaryl.

[0047] In some embodiments, the compound is a compound of formula (I). for , in R1 is C 1-4 alkoxy group; and R2 is a C group optionally substituted with one or more halogens. 1-4 Alkyl; or When R1 and R2 together form a fused 5- to 8-membered heterocyclic alkyl ring, a fused 5- to 6-membered heteroaryl ring, or a fused 7- to 15-membered spirocyclic heterocyclic alkyl ring system, or It is a 9- to 19-membered heterocyclic group, wherein the fused ring is optionally surrounded by halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl or 4- to 8-membered heterocyclic alkyl ring-substituted, wherein the C 1-4 Alkyl, the C 3-6 The cycloalkyl or the 4- to 8-membered heterocycloalkyl ring is optionally substituted with one or more halogens or C-membered heterocycloalkyl rings. 1-4 Alkyl substitution; R3 is a C-shaped element optionally substituted with one or more halogens. 1-4 alkyl.

[0048] In some embodiments, the compound is a compound of formula (I), wherein R1 is a methoxy group; R2 is fluorine; and R3 is a C group optionally substituted with one or more fluorine groups. 1-4 alkyl.

[0049] In some embodiments, the compound is a compound of formula (I), wherein for Among them, R4 and R 14 Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 Alkyl; X1 is CR 5a R 5b or NR 5c ;R 5a H or C optionally substituted with one or more halogens 1-4 Alkyl, and R 5b C 1-4 Alkyl or C 3-6 cycloalkyl, wherein the C 1-4 Alkyl or C 3-6 The cycloalkyl group is optionally substituted with one or more halogens; or R 5a and R 5b Together they form spirocyclic 3 to 8-membered heterocyclic alkyl or spirocyclic C 3-6 Cycloalkyl, wherein the 3- to 8-membered heterocycloalkyl or the C 3-6 The cycloalkyl group is optionally substituted with the following: C14 groups optionally substituted with one or more halogens. 1-4 Alkyl, 4- to 6-membered spirocyclic heterocyclic alkyl, or optionally C 1-4 Alkyl-substituted C 3-6 Spirocycloalkyl, the C 1-4 Alkyl groups may optionally be substituted with one or more halogens; R 5c C 1-4 Alkyl or C 3-6 cycloalkyl, wherein the C1-4 Alkyl or C 3-6 The cycloalkyl group is optionally substituted with one or more halogens; X2 is CR 6a R 6b -CH2CR 6a R 6b C=O, O or NR 6c ;R 6a And R 6b Each independently represents H and C. 1-4 Alkyl, 5- to 6-heteroaryl, 6-aryl, C 3-7 Cycloalkyl or 4- to 7-membered heterocycloalkyl, wherein the C 1-4 alkyl, the 5- to 6-membered heteroaryl, the 6-membered aryl, the C 3-8 The cycloalkyl group or the 4- to 7-membered heterocycloalkyl group is optionally converted by one or more halogens or C. 1-4 Alkyl substitution; R 6c C that is optionally substituted with one or more halogens 1-4 Alkyl; X3 is CR 7a R 7b , O or NR 7c ;R 7a and R 7b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 Alkyl; R 7c C that is optionally substituted with one or more halogens 1-4 Alkyl; X4 is CR 8a R 8b or NR 8c ;R 8a For H and R 8b C is a halogen or optionally substituted with one or more halogens. 1-4 Alkyl; R 8a and R 8b Together they form spirocyclic 3 to 8-membered heterocyclic alkyl or C 3-6 Cycloalkyl, wherein the 3- to 8-membered heterocycloalkyl or the C 3-6 The cycloalkyl group is optionally substituted with the following: C14 groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 8-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl, the 4 to 8-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group is optionally substituted with one or more halogens; and R 8c For H, C 1-4 Alkyl, 4- to 8-membered heterocyclic alkyl or C 3-6 cycloalkyl, wherein the C 1-4Alkyl, the 4- to 8-membered heterocyclic alkyl or the C 3-6 The cycloalkyl group is optionally substituted with one or more halogens; provided that neither X2 nor X3 can both contain O or N heteroatoms, and neither X3 nor X4 can both contain O or N heteroatoms.

[0050] In some embodiments, the compound is a compound of formula (I), wherein R 14 H is H; X1 is CR 5a R 5b ;R 5a Let H be the number of 'R', and R be the number of 'R'. 5b It is methyl or cyclopropyl.

[0051] In some embodiments, the compound is a compound of formula (I), wherein R4 is H, fluorine, or methyl; X1 is CR 5a R 5b ;R 5a For H, R 5b X2 is methyl; X3 is O and X4 is CR. 7a R 7b And R 7a and R 7b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 Alkyl; or X3 is O and X2 is CR 6a R 7b And R 6a and R 6b Each is independently H; and X4 is CR. 8a R 8b And R 8a and R 8b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 Alkyl, or R 8a and R 8b Together they form 4 to 7-membered heterocyclic alkyl rings or C 3-7 Cycloalkyl ring, wherein the 4- to 7-membered heterocyclic alkyl group or the C 3-7 The cycloalkyl group is optionally substituted with: halogen, C-aryl group optionally substituted with one or more halogens. 1-4 Alkyl, 4- to 7-membered spirocyclic heterocyclic alkyl, or optionally C 1-4 Alkyl-substituted C 3-7 Spirocycloalkyl, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

[0052] In some embodiments, the compound is a compound of formula (I), wherein for , or , where R 10aand R 10b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 Alkyl; or R 10a and R 10b Together they form a spiral ring C 3-6 Cycloalkyl or spirocyclic 6-membered heterocyclic alkyl; R 11a and R 11b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 Alkyl; R 12a and R 12b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 Alkyl; R 13a It is methyl, ethyl, or cyclopropyl; R 13b For H; and R 14 C is H, a halogen, or optionally substituted with one or more halogens. 1-4 alkyl.

[0053] In some embodiments, the compound is a compound of formula (I), wherein R 10a and R 10b Each is independently H or methyl; R 11a and R 11b Each is independently H, methyl, or ethyl; R 12a and R 12b Each is independently H or methyl; R 13a It is methyl, ethyl, or cyclopropyl; R 13b For H; and R 14 It can be H, fluorine, or methyl.

[0054] In some embodiments, the compound is a compound of formula (I), wherein R4 is H or fluorine; R 13a It is methyl, ethyl, or cyclopropyl; and R 14 For H.

[0055] In some embodiments, the compound is a compound of formula (I), wherein X2 is NR. 6c And R 6c C 1-4 alkyl.

[0056] In some embodiments, the compound is a compound of formula (I), wherein for , where R 13a It is H or methyl, and R4 is H, F or methyl.

[0057] In some embodiments, the compound is a compound of formula (I). for , or , where R4 is H, fluorine or methyl.

[0058] In some embodiments, the compound is a compound of formula (I), wherein R 10a R 10b and R 11a Each is either H or methyl.

[0059] In some embodiments, the compound is a compound of formula (I), wherein R 11b It can be H, methyl, or ethyl.

[0060] In some embodiments, the compound is a compound of formula (I), wherein for or R4 is H, a halogen, or a methyl group optionally substituted with one or more halogens; and R 9a and R 9b Each is independently H, a halogen, or a methyl group optionally substituted with one or more halogens.

[0061] In some embodiments, the compound is a compound of formula (I), wherein R4 is H, fluorine, or methyl; and R 9a and R 9b Each of the following is independently H, fluorine, chlorine, or a methyl group optionally substituted with one or more fluorine molecules.

[0062] In some embodiments, the compound is a compound of formula (I), wherein for R 20 H, halogen, C 1-4 Alkyl or C 1-4 alkoxy group, wherein the C 1-4 alkyl and the C 1-4 Each alkoxy group may optionally be substituted with one or more halogens; X5 is CR. 25a R 25b or NR 25c ;R 25a and R 25b Each is independently H or optionally substituted with one or more halogens or alkoxy groups. 1-4 Alkyl; or R 25a and R 25b Together they form C 3-6 Cycloalkyl rings or 4- to 6-membered heterocyclic rings, each optionally coated with halogen, C 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or spirocyclic 4- to 6-membered heterocyclic cyclic substitution, wherein the C 1-4 Alkyl, spirocyclic C 3-6Cycloalkyl or the spirocyclic 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens or C 1-4 Alkyl substitution; R 25c C 1-4 alkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 The alkyl group, the 5- to 6-membered heteroaryl group, or the C6 aryl group may optionally be affected by one or more halogens or C6 halogens. 1-4 Alkyl substitution; X6 is CR 26a R 26b Or C=O; R 26a and R 26b Each is independently H or optionally by one or more halogens or C 1-4 Alkyl-substituted C 1-4 Alkyl; X7 is CR 27a R 27b or NR 27c ;R 27a and R 27b Each of the C atoms is independently a carbon atom whose H is optionally substituted with one or more halogens or alkoxy groups. 1-4 Alkyl; R 27a and R 27b Together they form C 3-6 Cycloalkyl ring or 4- to 6-membered heterocyclic ring, wherein the C 3-6 The cycloalkyl ring or the 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens, one or more C-membered rings. 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or spirocyclic 4- to 6-membered heterocyclic cyclic substitution, wherein the C 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or the spirocyclic 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens or C 1-4 Alkyl substitution; and R 27c For H, C 1-4 Alkyl, C 3-6 Cycloalkyl, 4- to 7-membered heterocycloalkyl or 5- to 11-membered spirocyclic heterocycloalkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 Alkyl, the C 3-6 Cycloalkyl, the 4- to 6-membered heterocycloalkyl, the 5- to 11-membered spirocycloheterocycloalkyl, the 5- to 6-membered heteroaryl, or the C6 aryl, optionally with one or more halogens or C 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be substituted with one or more halogens; provided that only one of X5 and X7 contains an N heteroatom.

[0063] In some embodiments, the compound is a compound of formula (I), wherein R 20 H, halogen, C 1-4 Alkyl or C 1-4alkoxy group, wherein the C 1-4 alkyl and the C 1-4 The alkoxy group may optionally be substituted with one or more halogens; X5 is CR. 25a R 25b ;R 25a and R 25b Each is independently a methyl group; or R 25a and R 25b Together they form a cyclopropyl group; X6 is C=O; R 26a and R 26b Each is independently H or optionally by one or more halogens or C 1-4 Alkyl-substituted C 1-4 Alkyl; X7 is NR 27c And R 27c C 1-4 Alkyl, C 3-6 Cycloalkyl, 4- to 7-membered heterocycloalkyl, 5- to 11-membered spirocyclic heterocycloalkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 Alkyl, the C 3-6 Cycloalkyl, the 4- to 6-membered heterocycloalkyl, the 5- to 11-membered spirocycloheterocycloalkyl, the 5- to 6-membered heteroaryl, or the C6 aryl, optionally with one or more halogens or C 1-4 Alkyl substitution, the C 1-4 Alkyl groups may optionally be substituted with one or more halogens; In some embodiments, the compound is a compound of formula (I), R 27c C 1-4 Alkyl group. In some embodiments, the compound is a compound of formula (I), wherein R 27c It is a 5-membered heteroaryl group. In some embodiments, the compound is of formula (I), wherein R 27c C that is optionally substituted with one or more halogens 3-6 Cycloalkyl or C substituted with one or more halogens 1-4 Alkyl group. In some embodiments, the compound is a compound of formula (I), wherein R 27c C 1-4 Alkyl, 4- to 7-membered heterocyclic alkyl or 5- to 11-membered spirocyclic heterocyclic alkyl, wherein the C 1-4 Alkyl groups, the 4- to 7-membered heterocyclic alkyl groups, and the 5- to 11-membered spirocyclic heterocyclic alkyl groups are optionally oxidized by one or more halogens or C. 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

[0064] In some embodiments, the compound is a compound of formula (I), wherein for or , where R 20 It can be H, fluorine, methyl, ethyl, methoxy, -CH2-O-CH3, or -CF3. In some embodiments, the compound is a compound of formula (I), wherein R 27c It is a tetrahydropyran optionally substituted with one or more methyl groups or fluorine groups. In some embodiments, the compound is a compound of formula (I), wherein R 27c for or , where R 32a R 32b R 33a and R 33b Each is independently H, methyl, or fluorine; or R 32a and R 32b Or R 33a and R 33b One of them together forms a spiral ring C 3-6 Cycloalkyl or spirocyclic 3 to 6-membered heterocyclic rings.

[0065] In some embodiments, the compound is a compound of formula (I), wherein R 27c for In some embodiments, the compound is a compound of formula (I), wherein for , , or , or Where R 30a R 30b R 31a and R 31b Each is independently H, fluorine, or methyl; R 20 H, fluorine, methyl, ethyl, methoxy, -CH2-O-CH3, or -CF3; R 27c The methyl group is present. In some embodiments, the compound is a compound of formula (I), wherein R is a methyl group. 27c C 1-4 alkyl.

[0066] In some embodiments, the compound is a compound of formula (I), wherein for In some embodiments, the compound is a compound of formula (I), wherein for , where R 20 H, halogen, optionally C 1-4 alkoxy-substituted C 1-4 alkoxy groups, C groups optionally substituted with 4 to 6-membered heterocyclic groups or one or more halogens 1-4 alkyl.

[0067] In some embodiments, the compound is a compound of formula (I), wherein R 25c C 1-4 Alkyl; and R 27c To be optionally subjected to one or more halogens or C 1-4 alkoxy-substituted C 1-4 Alkyl, C 3-6 Cycloalkyl or 4- to 6-membered heterocycloalkyl, wherein the C 3-6 Cycloalkyl or 4- to 6-membered heterocycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

[0068] In some embodiments, the compound is a compound of formula (I), wherein R 27c C 1-4 Alkyl; and R 25c To be optionally subjected to one or more halogens or C 1-4 alkoxy-substituted C 1-4 Alkyl, C 3-6 Cycloalkyl or 4- to 6-membered heterocycloalkyl, wherein the C 3-6 Cycloalkyl or the 4- to 6-membered heterocycloalkyl group optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

[0069] In some embodiments, the compound is a compound of formula (I), wherein R 20 H or methyl. In some embodiments, the compound is a compound of formula (I), wherein... or ;where R 20 For H, halogen, optional C 1-4 alkoxy-substituted C 1-4 Alkoxycarbonyl, C substituted with a 4- to 6-membered heterocyclic group 1-4 Alkyl or one or more halogens; R 25c It is C 1-4 Alkyl; R 27c It is a 4- to 6-membered heterocyclic group or C 3-6 cycloalkyl, wherein the 4- to 6-membered heterocyclic group or the C 3-6 Cycloidal groups can be selectively C 1-4 Alkyl substitution.

[0070] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0071] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0072] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0073] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0074] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0075] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0076] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0077] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0078] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0079] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0080] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0081] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0082] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0083] In some embodiments, the compound has the chemical formula […]. Compounds or their pharmaceutically acceptable salts.

[0084] In some embodiments, the compound is selected from... Figure 1 The compounds are those listed in Table 3 or their pharmaceutically acceptable salts. In some embodiments, the compounds are selected from the compounds listed in Table 3 or their pharmaceutically acceptable salts.

[0085] definition For convenience, certain terms used in this specification, embodiments, and appended claims are collected herein before further description of the invention. These definitions should be read in light of the remainder of this disclosure and are understood by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0086] To make the invention easier to understand, certain terms and phrases will be defined below and throughout the specification.

[0087] The articles “a” and “an” are used in this text to refer to one or more of the grammatical objects of the article (i.e., at least one). For example, “an element” refers to one or more elements.

[0088] The phrase “and / or” as used herein in the specification and claims should be understood to mean “any one or both” of the elements so connected, that is, elements that exist jointly in some cases and separately in others. Multiple elements listed using “and / or” should be interpreted in the same way, that is, “one or more” of the elements so connected. Optionally, other elements may exist besides those specifically identified by the “and / or” entry, whether related to or unrelated to the specifically identified element. Thus, as a non-limiting example, references to “A and / or B” used in conjunction with open-ended language such as “comprising” may, in one embodiment, refer only to A (optionally including elements other than B); in another embodiment, refer only to B (optionally including elements other than A); in yet another embodiment, refer to both A and B (optionally including other elements); and so on.

[0089] The word “or” as used in this specification and claims shall be understood to have the same meaning as “and / or” as defined above. For example, when items in a list are separated, “or” or “and / or” shall be interpreted as inclusive, that is, including multiple elements or at least one of the elements in the list, and including more than one of them, and optionally including additional unlisted items. Terms that clearly indicate the opposite, such as “only one of…” or “exact one of…” or, when used in a claim, “consisting of…”, will refer to including multiple elements or exactly one of the elements in the list. Generally, as used herein, the term “or” shall be interpreted as indicating an exclusive alternative (i.e., “one of… or the other, but not both”) only when preceded by an exclusive term such as “any one of…”, “one of…”, “only one of…”, or “exact one of…”. When used in a claim, “consisting substantially of…” shall have its ordinary meaning as used in the field of patent law.

[0090] The term “at least one” as used in this specification and claims, when referring to a list of one or more elements, should be understood to mean at least one element selected from any one or more elements in the list of elements, but does not necessarily include at least one of each element specifically listed in the list of elements and does not exclude any combination of elements in the list of elements. This definition also allows for the optional presence of elements other than those specifically identified in the list of elements referred to by the term “at least one,” whether related to or unrelated to those specifically identified elements. Thus, as a non-limiting example, “at least one of A and B” (or equivalently, “at least one of A or B”, or equivalently, “at least one of A and / or B”) in one embodiment may refer to at least one, optionally including more than one A, without B (and optionally including elements other than B); in another embodiment, it may refer to at least one, optionally including more than one B, without A (and optionally including elements other than A); in yet another embodiment, it may refer to at least one, optionally including more than one A, and at least one, optionally including more than one B (and optionally including other elements); and so on.

[0091] It should also be understood that, unless explicitly indicated to the contrary, in any method claimed herein that includes more than one step or operation, the order of the steps or operations of the method is not necessarily limited to the order in which the steps or operations of the method are described.

[0092] In the claims, and in the foregoing description, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “possessing,” “constituting of,” and similar phrases shall be understood as open-ended, i.e., meaning including but not limited to. As set forth in Section 2111.03 of the United States Patent Office Manual of Patent Examining Procedures, only the transitional phrases “constituting of” and “substantially consisting of” shall be closed or semi-closed transitional phrases, respectively.

[0093] Certain compounds contained in the compositions of the present invention may exist in specific geometric or stereoisomeric forms. Furthermore, the polymers of the present invention may also be optically active. The present invention contemplates all such compounds within its scope, including cis and trans isomers, R-enantiomers and S-enantiomers, diastereomers, (d)-isomers, (l)-isomers, racemic mixtures thereof, and other mixtures thereof. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are intended to be included in the present invention.

[0094] For example, if a specific enantiomer of the compound of the present invention is required, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary agent, wherein the resulting mixture of diastereomers is separated and the auxiliary groups are cleaved to provide the desired enantiomer in pure form. Alternatively, in the case where the molecule contains a basic functional group such as an amino group or an acidic functional group such as a carboxyl group, diastereomeric salts are formed with a suitable optically active acid or base, and the resulting diastereomers are then resolved by stepwise crystallization or chromatographic methods known in the art, and the pure enantiomers are subsequently recovered.

[0095] The structures described herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds produced by replacing hydrogen with deuterium or tritium, or by replacing carbon with carbon enriched in 13C or 14C, are within the scope of this invention.

[0096] As used herein, the phrase “pharmaceuticalally acceptable excipient” or “pharmaceuticalally acceptable carrier” refers to a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, that participates in the transport or delivery of the subject chemical substance from one organ or part of the body to another organ or part of the body. Each carrier must be “acceptable” in the sense that it is compatible with other components of the formulation, harmless to the patient, and substantially non-pyrogenic. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution. (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic and compatible substances used in the formulation. In some embodiments, the pharmaceutical compositions of the present invention are pyrogen-free, i.e., do not induce a significant increase in temperature when administered to a patient.

[0097] The term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic and organic acid addition salt of one or more compounds. These salts can be prepared in situ during the final separation and purification of one or more compounds, or by reacting one or more purified compounds in free base form with a suitable organic or inorganic acid and separating the resulting salt. Representative salts include hydrobromide, hydrochloride, sulfate, hydrogen sulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, methanesulfonate, glucono-p-ethyl, lactobionate, and laurylsulfonate, etc. (See, for example, Berge et al. (1977), "Pharmaceutical Salts," J. Pharm. Sci. 66:1-19.)

[0098] In other cases, the compounds used in the methods of this invention may contain one or more acidic functional groups and are therefore capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term "pharmaceutically acceptable salt" in these cases refers to a relatively non-toxic inorganic and organic base addition salt of (one or more) compounds. These salts may also be prepared in situ during the final separation and purification of (one or more) compounds, or by reacting (one or more) purified compounds in free acid form with suitable bases, such as hydroxides, carbonates, or bicarbonates of pharmaceutically acceptable metal cations, with ammonia, or with pharmaceutically acceptable primary, secondary, or tertiary organic amines. Representative alkali metal or alkaline earth metal salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts, etc. Representative organic amines that can be used to form base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc. (see, for example, Berge et al., above).

[0099] The "therapeutic effective amount" (or "effective amount") of a compound used for treatment refers to the amount of a compound in a formulation that, when administered as part of a desired dosage regimen (to mammals, preferably humans), relieves symptoms, improves condition, or slows the onset of disease symptoms according to clinically acceptable criteria for the treatment of a disease or condition or for cosmetic purposes, such as a reasonable benefit / risk ratio applicable to any medical treatment.

[0100] The term "preventive or therapeutic" treatment is recognized in the art and includes the administration of one or more of the subject composition to a host. Treatment is preventive (i.e., it protects the host from developing the unwanted condition) if it is administered before the clinical manifestation of an unwanted symptom (e.g., disease or other unwanted condition in the host animal), and therapeutic (i.e., it is intended to reduce, improve, or stabilize the existing unwanted symptom or its side effects) if it is administered after the manifestation of the unwanted symptom.

[0101] The term "patient" refers to a mammal that requires specific treatment. In some implementations, the patient is a primate, dog, cat, or horse. In some implementations, the patient is a human.

[0102] Whenever a term (e.g., alkyl or aryl) or any of its prefix roots (e.g., alkane- or aryl-) appears in the name of a substituent, the name should be interpreted to include the limitations provided herein. For example, attaching the suffix "-ene" to a group indicates that the group is a divalent moiety, such as arylene being a divalent aryl moiety, heteroarylene being a divalent heteroaryl moiety, and heterocyclic alkyl being a divalent heterocyclic alkyl moiety. Similarly, attaching the suffix "-oxy" to a group indicates that the group is attached to the parent molecule structure via an oxygen atom (-O-), as used herein, such as "alkyloxy," "alkoxy," or "cycloalkoxy."

[0103] Aliphatic chains include the alkyl, alkenyl, and alkynyl categories as defined below. Straight-chain aliphatic chains are limited to the unbranched carbon chain portion. As used herein, the term "aliphatic group" refers to a straight-chain, branched, or cyclic aliphatic hydrocarbon group, and includes saturated and unsaturated aliphatic groups such as alkyl, alkenyl, or alkynyl groups.

[0104] "Alkyl" refers to a fully saturated cyclic or acyclic, branched or unbranched carbon chain moiety having a specified number of carbon atoms, or, if not specified, 1 to up to 30 carbon atoms. For example, an alkyl group having 1 to 8 carbon atoms refers to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl, as well as those moieties that are positional isomers of these moieties. Alkyl groups having 10 to 30 carbon atoms include decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecanyl, octadecyl, nonadecanyl, eicosyl, dodecyl, tridecyl, and tetradecyl. In some embodiments, straight-chain or branched alkyl groups have 30 or fewer carbon atoms in their main chain (e.g., for straight-chain C1-C2). 30 For C3-C branches 30 (and more preferably has 20 or fewer carbon atoms. The alkyl group may be substituted or unsubstituted. As used herein, "Me" and -CH3 both refer to methyl.)

[0105] As used herein, the term "alkylene" refers to an alkyl group having a specified number of carbon atoms, such as 2 to 12, containing two connection sites on its longest carbon chain that connect to the rest of the compound. Non-limiting examples of alkylenes include methylene-(CH2)-, ethylene-(CH2CH2)-, n-propylene-(CH2CH2CH2)-, isopropylene-(CH2CH(CH3))-, etc. Alkylenes can be cyclic or acyclic, branched or unbranched carbon chain portions, and may optionally be substituted with one or more substituents.

[0106] "Cycloalkyl" refers to a monocyclic, bicyclic, bridged, spirocyclic, or polycyclic saturated carbocyclic ring, each having 3 to 12 carbon atoms. Similarly, preferred cycloalkyl groups have 3-10 carbon atoms in their ring structure, and more preferably 3-6 carbon atoms. Cycloalkyl groups may be substituted or unsubstituted. Exemplary cycloalkyl groups include cyclopropyl (C3), cyclobutyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cycloheptyl (C7), and cyclooctyl (C8).

[0107] Unless otherwise specified, "lower alkyl" as used herein refers to an alkyl group as defined above, having one to ten carbon atoms, more preferably one to six carbon atoms, in its main chain structure, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Similarly, "lower alkenyl" and "lower alkynyl" have similar chain lengths. Throughout this application, preferred alkyl groups are lower alkyl groups. In some embodiments, substituents referred to herein as alkyl groups are lower alkyl groups.

[0108] As used herein, the term "aryl" includes 3 to 12-membered monocyclic aromatic groups, substituted or unsubstituted, wherein each atom in the ring is carbon (i.e., carbocyclic aryl) or one or more of the atoms are heteroatoms (i.e., heteroaryl). Preferably, aryl comprises 5 to 12-membered rings, more preferably 6 to 10-membered rings. The term "aryl" also includes polycyclic ring systems having two or more cyclic rings, wherein two or more carbons are shared by two adjacent rings, and wherein at least one ring is an aromatic ring, for example, other rings may be cycloalkyl, cycloalkenyl, cycloynyl, aryl, heteroaryl, and / or heterocyclic. Carbocyclic aryl includes benzene, naphthalene, phenanthrene, phenol, aniline, and similar groups. Heteroaryl includes substituted or unsubstituted aromatic 3 to 12-membered ring structures, more preferably 5 to 12-membered rings, more preferably 5 to 10-membered rings, whose ring structures include one to four heteroatoms. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Aryl and heteroaryl groups can be monocyclic, bicyclic, or polycyclic.

[0109] As used herein, the terms "halo," "halide," or "halogen" refer to halogens and include, but are not limited to, fluorine, chlorine, bromine, iodine, etc., in radioactive and non-radioactive forms. In a preferred embodiment, the halogen is selected from fluorine, chlorine, and bromine.

[0110] The term "heterocyclyl" or "heterocyclic group" refers to a 3- to 12-membered ring structure, more preferably a 5- to 12-membered ring, and even more preferably a 5- to 10-membered ring, whose ring structure includes one to four heteroatoms. The heterocycle can be monocyclic, bicyclic, spirocyclic, or polycyclic. Heterocyclic groups include, for example, thiophene, thiamethoxam, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indazine, isoindole, indole, indazole, purine, quinazine, isoquinoline, quinoline, phthalazine, naphthidine, quinoxaline, quinazolin, quinazolin, cyclophosphine, pteridine, carbazole, caroline, phenanthridine, acridine, pyrimidine, phenanthrene-rholine. Phenazine, phenarsazine, phenothiazine, furazine, phenoxazine, pyrrolidine, oxacyclopentane, thiocyclopentane, oxazole, piperidine, piperazine, morpholine, lactone, lactam (such as azacyclobutanone and pyrrolidone), sulfonolactam, sulfonolactone, etc. The heterocycle may be substituted at one or more positions with the substituents described above, such as halogens, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, mercapto, imino, amide, phosphate ester, phosphonate, hypophosphonate, carbonyl, carboxyl, silyl, aminosulfonyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, -CF3, -CN, etc.

[0111] As used herein, the term "heterocyclic alkyl" is a non-aromatic heterocyclic group in which at least one atom is a heteroatom, such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus, and the remaining atoms are carbon. Examples of heterocyclic alkyl groups are pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiophenyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazine, azaheterocyclic butyl, oxacyclocyclic butyl, thiocyclic butyl, high-piperidinyl, oxacycloheptyl, thiocyclic heptyl, and oxacycloheptyl. Alkyl groups, diazacycloheptyl groups, thiazacycloheptyl groups, 1,2,3,6-tetrahydropyridinyl groups, 2-pyrrolinyl groups, 3-pyrrolinyl groups, dihydroindolyl groups, 2H-pyranyl groups, 4H-pyranyl groups, dioxanecyclohexyl groups, 1,3-dioxanecyclopentyl groups, pyrazolinyl groups, dioxanecyclohexyl groups, 3-azabicyclo[3.1.0]hexyl groups, 3-azabicyclo[4.1.0]heptyl groups, 3H-indolyl groups, and quinazinyl groups. Heterocyclic alkyl groups may be substituted or unsubstituted, as described, for example, with respect to the heterocyclic groups described herein.

[0112] The term "carbonyl" is generally accepted in the art and includes portions that can be represented by the following formula: , Where X' is a bond or represents oxygen or sulfur, and R 15 Indicates hydrogen, alkyl, alkenyl, -(CH2) m -R 10 Or a pharmaceutically acceptable salt, R 16 Indicates hydrogen, alkyl, alkenyl, or -(CH2). m -R 10 , where m and R 10 As defined above. When X' is oxygen and R 15 Or R 16 When X' is not hydrogen, this formula represents "ester". When X' is oxygen and R... 15 As defined above, this part is referred to herein as the carboxyl group, and especially when R 15 When X' is hydrogen, this formula represents "carboxylic acid". When X' is oxygen and R is oxygen, the formula represents "carboxylic acid". 16 When X' is hydrogen, the formula represents "formate ester". On the other hand, when X' is a bond and R... 15 When the hydrogen atom is not hydrogen, the above formula represents a "ketone" group. When X' is a bond and R... 15 When the hydrogen is present, the above formula represents an "aldehyde" group.

[0113] As used herein, the term "substituted" is intended to include all permissible substituents of an organic compound. In a broad aspect, the permissible substituent includes acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of the organic compound. Illustrative substituents include, for example, the substituents described above, and are substituted, for example, by one or more substituents selected from: alkyl, cycloalkyl, heterocyclic alkyl, halogen, OH, OMe, C(H)F2, C(F)H2, CF3, C(H)2CF3, SF5, CHFCH2amine, CH2amine, and CN. For a suitable organic compound, the permissible substituent may be one or more and may be the same or different. For the purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituent of the organic compound described herein that satisfies heteroatom valences. This invention is not intended to be limited in any way to the permissible substituents of organic compounds. It should be understood that “substitution” or “being substituted” includes implied limitations, namely, that the substitution is based on the permissible valence of the substituted atom and the substituent, and that the substitution produces a stable compound, for example, a compound that does not spontaneously undergo transformations, such as transformations by rearrangement, cyclization, elimination, etc.

[0114] As used herein, the term “nitro” refers to -NO2; the term “halogen” refers to -F, -Cl, -Br or -I; the term “hydroxyl” refers to -OH; and the term “cyano” refers to -CN.

[0115] As used herein, when the definitions of, for example, alkyl, m, n, etc., appear more than once in any structure, their intent is unrelated to their definitions elsewhere in the same structure.

[0116] As used herein, the term "prodrug" encompasses a compound that is converted into a therapeutically active agent under physiological conditions. Common methods for manufacturing prodrugs involve hydrolysis under physiological conditions to reveal selected moieties of the desired molecule. In other embodiments, the prodrug is converted in vivo by the enzymatic activity of a host animal. Therefore, a prodrug includes a compound converted in vivo to obtain the disclosed compound or any other pharmaceutically acceptable form of the compound. In embodiments, the prodrug may be inactive when administered to a subject, but can be converted in vivo, for example, by hydrolysis, into the active compound. See, for example, Bundgard, H. 前药设计 (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam). Discussion of prodrugs is provided in Higuchi, T. et al., “Pro-drugsas Novel Delivery Systems”. 美国化学会 专题讨论会系列 Volume 14; and 药物设计中的生物可逆载体 Edward B. Roche (ed.), American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference in their entirety. Prodrugs can typically be processed using well-known methods, such as... 伯格药物化学 与药物发现 , 172-178, 949-982 (edited by Manfred E. Wolff, 5th edition, 1995); and 前药设计 Prepared by the method described in (H. Bundgaard, ed., Elselvier, New York, 1985). The term "prodrug" also refers to any covalently bonded carrier that releases the active compound in vivo when administered to a subject.

[0117] Prodrugs of the compounds described herein can be prepared by modifying functional groups present in the active compound in a manner that allows the modification to be performed conventionally or cleaved in vivo to provide the compound described herein (i.e., the parent active compound). Prodrugs include compounds in which a hydroxyl, amino, or thiol group is bonded to any group that, when administered to a subject as a prodrug of the active compound, cleaves to form a free hydroxyl, free amino, or free thiol group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of the alcohol functional group in the active compound, or acetamide, formamide, and benzamide derivatives of the amine functional group in the active compound. Other examples of prodrugs include compounds containing a —NO, —NO2, —ONO, or —ONO2 moiety.

[0118] For the purposes of this invention, chemical elements are identified according to the CAS version of the periodic table (Hand book of Chemistry and Physics, 67th edition, 1986-87).

[0119] In some embodiments, the compound is selected from... Figure 1 The compound or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is selected from the compounds in Table 3 or a pharmaceutically acceptable salt thereof.

[0120] Table 3. Exemplary Compounds .

[0121] Therapeutic uses of compounds of formula (I) In tissues, normal fibroblasts exist in a low population of only 4-5%. However, during fibrosis, they proliferate and can account for 80-90% of organ mass. Myofibroblasts in fibrotic tissue produce large amounts of extracellular matrix proteins, leading to tissue scarring and loss of function. Inhibition of myofibroblasts can counteract these processes. Integrins promote cell proliferation, survival, hypertrophic growth, and fibrosis. As described in this article, integrin inhibition modulates these key factors, leading to the progression of pulmonary hypertension (e.g., PAH).

[0122] This invention provides a method for treating a disease associated with enhanced integrin α5β1 expression or activity, comprising administering an integrin α5β1 inhibitor. In one aspect, the invention provides an integrin α5β1 inhibitor for treating a subject in need of such treatment with a disease associated with enhanced integrin α5β1 expression or activity, comprising administering the integrin α5β1 inhibitor and a pharmaceutical excipient to the subject.

[0123] In some embodiments, the disease is heart failure or right ventricular failure. In one aspect, the invention provides a method of treating a subject with pulmonary arterial hypertension (PAH), comprising administering an integrin α5β1 inhibitor. In some embodiments, the integrin α5β1 inhibitor is administered orally, intravenously, subcutaneously, intranasally, percutaneously, intraperitoneally, intramuscularly, or intrapulmonaryly. A method of treating a subject suffering from fibrosis or fibrosis-related conditions is also provided, comprising administering to said subject a therapeutically effective amount of the integrin α5β1 inhibitor according to the invention. As used herein, the term "fibrosis" refers to a condition characterized by the deposition of extracellular matrix components in the skin or organs (including the lungs, kidneys, heart, liver, skin, and joints), resulting in scar tissue. The term also refers to the process of scar tissue formation.

[0124] In some implementations, fibrosis-related conditions are conditions or symptoms that can occur due to or are related to fibrosis. In some implementations, fibrosis and / or fibrosis-related conditions are selected from the following diseases or symptoms: renal fibrosis, liver fibrosis, cirrhosis, pulmonary fibrosis, skin fibrosis, biliary fibrosis, peritoneal fibrosis, myocardial fibrosis, pancreatic fibrosis, bone marrow and / or myelofibrosis, reperfusion injury after liver or kidney transplantation, interstitial lung disease (ILD), cystic fibrosis (CF), atherosclerosis, systemic sclerosis, osteosclerosis, herniated discs and other spinal cord injuries, fibromatosis, fibromyalgia, arthritis, restenosis. Pulmonary fibrosis includes idiopathic pulmonary fibrosis and scleroderma pulmonary fibrosis.

[0125] The methods described herein can be applied to in vivo or ex vivo cell populations. "In vivo" means within a living individual, such as an animal or human. In this case, the methods described herein can be used therapeutically within the individual. "Ex vivo" means outside a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples, including fluid or tissue samples obtained from an individual.

[0126] Information gathered from such uses can be used for experimental or clinical purposes to establish in vivo treatment regimens. The selected compounds can be further characterized to examine safety or tolerable doses in human or non-human subjects. Such properties can be examined using methods commonly known to those skilled in the art.

[0127] In some embodiments, the compounds described herein, or pharmaceutically acceptable salts, stereoisomers, mixtures of stereoisomers, tautomers, or deuterated analogs thereof, may be used to treat subjects who have or are suspected of having a disease state, condition, or symptom that is responsive to or believed to be responsive to inhibition of α5β1 integrin activity (collectively, the “Indications”). In some embodiments, the compounds described herein may be used to inhibit the activity of α5β1 integrin. In some embodiments, this disclosure provides compounds described herein that can be used as inhibitors of α5β1 integrin. In some embodiments, this disclosure provides a method of treating or preventing an inflammatory disease or symptom comprising administering the compounds described herein to a patient. In some embodiments, this disclosure provides a pharmaceutical composition comprising the compounds described herein and a pharmaceutically acceptable carrier. In some embodiments, the compounds described herein are provided for treating patients with inflammatory diseases or symptom mediated at least in part by α5β1 integrin.

[0128] Co-administration may also include administering component drugs, such as one or more compounds described herein, and one or more additional (e.g., second, third, fourth, or fifth) therapeutic agents. Such combinations of one or more compounds described herein and one or more additional therapeutic agents may be administered simultaneously or sequentially (one after another) at reasonable intervals of administration (e.g., from about 1 minute to 24 hours) based on the pharmacokinetic and / or pharmacodynamic properties of the individual agents or the combination. Co-administration may also involve treatment with a fixed combination, wherein the agents in the treatment regimen may be combined in a fixed dose or combined dose medium (e.g., solid, liquid, or aerosol).

[0129] Treatment with a second, third, fourth, or fifth active agent may be performed before, simultaneously with, or after treatment with the compounds described herein. In some embodiments, the compounds described herein are combined with another active agent in a single dosage form.

[0130] In the implementation plan, the subjects are adult subjects.

[0131] Dosage and frequency In some implementations, the compound of formula (I) described herein or any pharmaceutically acceptable salt thereof is administered to a patient in need in an effective amount.

[0132] In some embodiments, a method of treating diseases associated with enhanced integrin α5β1 expression or activity includes administering an integrin α5β1 inhibitor at a dose from 1 mg / kg to 1000 mg / kg. In some embodiments, the dose is at least 2 mg / kg, at least 4 mg / kg, at least 6 mg / kg, or at least 8 mg / kg. In some embodiments, the dose is at least 10 mg / kg, at least 20 mg / kg, at least 30 mg / kg, at least 40 mg / kg, at least 50 mg / kg, at least 60 mg / kg, at least 70 mg / kg, at least 80 mg / kg, at least 90 mg / kg, or at least 100 mg / kg. In some embodiments, the dose is at least 200 mg / kg, at least 300 mg / kg, at least 400 mg / kg, at least 500 mg / kg, at least 600 mg / kg, at least 700 mg / kg, at least 800 mg / kg, at least 900 mg / kg, or at least 1000 mg / kg.

[0133] In one implementation scheme, the subject receives a dose of compound (I) or a pharmaceutically acceptable salt thereof once daily. In another implementation scheme, the subject receives a dose of compound (I) or a pharmaceutically acceptable salt thereof twice daily.

[0134] cardiac function parameters To determine the efficacy of the therapy of this invention, various endpoint parameters can be assessed, such as α5β1 level, pulmonary vascular resistance (PVR), mean pulmonary artery pressure (PAP), cardiac index (CI), mean pulmonary capillary wedge pressure (PCWP), right atrial pressure (RAP), six-minute walk distance (6MWD), brain natriuretic peptide (BNP) level, lung diffusion capacity (DLCO), and death or survival. See Chung et al., Chest (2010), 138(6):1383-1394.

[0135] PVR is commonly used as an endpoint parameter to determine the therapeutic efficacy of PAH. A PVR > 240 dyn·sec / cm5 indicates mild PAH. A PVR of 600-800 dyn·sec / cm5 indicates moderate to severe PAH. After treatment with the methods and compositions of the present invention, a PVR reduction of 130 dyn·sec / cm5 or more indicates effective treatment. For example, administration of an A5B1 inhibitor to a subject with PAH results in a PVR reduction of 180-350 dyn·sec / cm5, indicating effective treatment.

[0136] Mean pulmonary artery pressure (PAP) is also used as an endpoint parameter to determine the efficacy of treatment for PAH. Subjects without PAH have a mean PAP of approximately 15–24 mmHg. Subjects with mild PAH have a mean PAP of approximately 25–30 mmHg (e.g., >25 mmHg at rest or 30 mmHg during exercise). Subjects with severe PAH have a PAP greater than 30 mmHg, such as 40–70 mmHg or 60–70 mmHg. A PAP reduction greater than 1.5 mmHg after treatment indicates effective treatment. In some implementations, a PAP reduction greater than 5, 10, 20, 40, or 50 mmHg indicates effective treatment.

[0137] The cardiac index (CI) is also used as an endpoint parameter to determine the efficacy of treatment for PAH. A low CI or a decreased CI indicates heart failure. For example, a CI of 2.5 L / min / m2 or less indicates PAH or heart failure. After treatment, an increase in CI greater than 0.3 L / min / m2 indicates effective treatment.

[0138] Mean pulmonary capillary wedge pressure (PCWP) can be used as an endpoint parameter to determine the efficacy of treatment for PAH. A mean PCWP less than or equal to 18 mmHg (e.g., less than or equal to 10 mmHg) indicates that the subject has PAH. An increase in mean PCWP greater than 0.5 mmHg after treatment indicates that the treatment is effective.

[0139] Right atrial pressure (RAP) is also used as an endpoint parameter to determine the efficacy of treatment for PAH. Subjects without PAH have a normal RAP of 0–8 mmHg. A RAP of 8 mmHg or greater indicates PAH. Subjects with severe PAH have a RAP of approximately 20 mmHg. A decrease in RAP greater than 0.5 mmHg after treatment indicates effective treatment.

[0140] The six-minute walking distance (6 MWD) was used as the endpoint parameter to determine the efficacy of PAH treatment. The mean 6 MWD for patients with CTD-PAH was approximately 300 m. An increase of 25 m or more or a greater than 10% in 6 MWD after treatment indicated effective treatment. For example, a 6 MWD of 1000 m or more after treatment indicated effective treatment.

[0141] Brain natriuretic peptide (BNP) is used as an endpoint parameter to determine the efficacy of treatment for PAH. BNP is a sensitive marker of worsening heart failure and a predictor of mortality in PAH patients. Normal BNP levels are <100 pg / mL, e.g., 30–90 pg / mL. Higher BNP levels indicate worsening heart failure. BNP levels of approximately 100–200 pg / mL (e.g., 160 pg / mL or higher) indicate early heart failure. BNP levels of approximately 200–1000 pg / mL indicate true heart failure. The mean BNP level in CTD-PAH patients is approximately 430 pg / mL. Any decrease in BNP levels after treatment indicates treatment efficacy.

[0142] N-terminal probrain natriuretic peptide (NT-proBNP): A reproducible, non-invasive parameter available for patients with PAH. BNP is produced in the ventricles of the heart and is elevated in PPH / IPAH. BNP levels have recently been shown to be closely associated with functional impairment in PPH / IPAH patients and matched with the degree of pulmonary hemodynamic changes and right-sided heart failure. BNP levels correlate longitudinally with functional assessments performed during the study. Plasma NT-pro-BNP was measured using a sandwich immunoassay (Elecsys analyzer, Roche Diagnostics, Manheim, Germany) using polyclonal antibodies that identify epitopes located in the N-terminal fragments (1 to 76) of pro-BNP (1 to 108).

[0143] Lung diffusion capacity (DLCO), or CO diffusion capacity, is also used as an endpoint parameter to determine the efficacy of treatment for PAH. DLCO measures the ability of carbon monoxide (CO) to diffuse across the membrane. Subjects without PAH have a normal DLCO greater than 80%. Subjects with PAH have an abnormal DLCO less than 80%, less than 65%, or less than 45%. Any increase in %DLCO after treatment indicates treatment efficacy.

[0144] In some implementations, administration of an integrin α5β1 inhibitor modulates the levels of biomarkers in subjects, wherein the biomarkers are selected from survivin, PCNA, Ki67, and annexin V.

[0145] Pulmonary hypertension Pulmonary hypertension (PH) is a syndrome characterized by increased pulmonary artery pressure. PH is hemodynamically defined as a systolic pulmonary artery pressure greater than 30 mmHg or a mean pulmonary artery pressure greater than 25 mmHg. See Zaiman et al., Am. J. Respir. Cell Mol. Biol. 33:425-31 (2005). Furthermore, due to the increased pressure, PH damages both the large and small pulmonary arteries. The walls of the smallest vessels thicken and they are no longer able to properly transfer oxygen and carbon dioxide between the blood and the lungs. Over time, pulmonary hypertension leads to thickening of the pulmonary arteries and narrowing of the passageway through which blood flows. Once pulmonary hypertension develops, the right side of the heart works harder to compensate; however, the increased effort causes it to become dilated and thickened. The proliferation of smooth muscle and endothelial cells, which are normally at rest, leads to vascular remodeling and occlusion of the pulmonary vessel lumen. This results in a progressive increase in pulmonary pressure because the luminal area through which blood is pumped decreases. Because blood tends to pool in the ventricles and legs, right ventricular enlargement puts the subject at risk of pulmonary embolism. If a clot forms in the pooled blood, it can eventually travel to and lodge in the lungs. The progressive increase in pressure also places an extra workload on the right ventricle, eventually leading to right ventricular failure and premature death in these patients.

[0146] Pulmonary arteries undergo a variety of pathological changes due to pulmonary hypertension (PH). Persistent vasoconstriction and structural remodeling of pulmonary vessels are fundamental characteristics of PH. Pulmonary vascular smooth muscle cells undergo a phenotypic shift from a normal contractile phenotype to a synthetic phenotype, leading to cell growth and matrix deposition. Histological examination of tissue samples from patients with pulmonary hypertension reveals intimal thickening and smooth muscle cell hypertrophy, especially in vessels with a diameter <100 μm. Furthermore, abnormal smooth muscle cells often overexpress endothelin and serotonin transporters, which may play a role in the development of PH.

[0147] The most common initial symptom of pulmonary hypertension is shortness of breath after exertion. Some people experience dizziness or fatigue after exertion, and colicky chest pain is common. General weakness is another problem because the body tissues are not receiving enough oxygen. Underlying lung disease can cause other symptoms such as cough and wheezing. Edema, especially in the legs, can occur because fluid can leak from veins into tissues, indicating pulmonary heart disease. Some people with pulmonary hypertension also have connective tissue disorders, particularly scleroderma. When both pulmonary hypertension and connective tissue disorders are present, Raynaud's phenomenon often precedes the onset of symptoms of pulmonary hypertension, sometimes by several years.

[0148] Treatments for some types of pulmonary hypertension often target the underlying lung disease. Currently, treatment options available to patients with PH target cellular dysfunctions that cause vasoconstriction. Therapies such as prostanoids, phosphodiesterase-5 inhibitors, and endothelin receptor antagonists primarily work by causing pulmonary vasodilation. Vasodilators, such as calcium channel blockers, nitric oxide, and prostacyclin, are often helpful for pulmonary hypertension associated with scleroderma, chronic liver disease, and HIV infection. In contrast, these drugs have not been proven effective for people with pulmonary hypertension due to an underlying lung disease. For most people with pulmonary hypertension of unknown cause, vasodilators (such as prostacyclin) significantly reduce pulmonary artery blood pressure. Intravenous administration of prostacyclin via a surgically implanted catheter under the skin improves quality of life, increases survival, and reduces the urgency of lung transplantation. Unfortunately, many patients respond poorly to these therapies or cease responding over time. At this point, the only remaining option is single or double lung transplantation to treat PH. Although some evidence suggests that available therapies have secondary effects on vascular remodeling, there are currently no therapies that target the abnormal cell proliferation in PAH.

[0149] In some embodiments, pulmonary hypertension is pulmonary venous hypertension (PVH). In some embodiments, PVH is attributed to left-sided heart failure. In some embodiments, pulmonary hypertension is pulmonary hypertension associated with respiratory symptoms and / or hypoxia. In some embodiments, pulmonary hypertension is pulmonary hypertension attributed to chronic thrombotic and / or embolic diseases. In some embodiments, pulmonary hypertension is other types of pulmonary hypertension. In some embodiments, other types of pulmonary hypertension are associated with: sarcoidosis, eosinophilic granuloma, histiocytosis X, lymphangiolomyiomatosis, or pulmonary vascular compression (e.g., adenopathy, tumors, or fibrosing medianstinitis). In some embodiments, pulmonary hypertension is associated with chronic obstructive pulmonary disease (COPD). In some embodiments, pulmonary hypertension is associated with pulmonary fibrosis. In some embodiments, pulmonary hypertension is associated with cardiac fibrosis. In some embodiments, pulmonary hypertension is early-stage or late-stage pulmonary hypertension.

[0150] In some embodiments, one or more symptoms of pulmonary hypertension are improved. In some embodiments, pulmonary hypertension is delayed. In some embodiments, pulmonary hypertension is prevented. In some embodiments, the treatments provided herein reduce pulmonary pressure. In some embodiments, the treatments provided herein inhibit and / or reduce the abnormal cell proliferation in the pulmonary arteries.

[0151] In some implementations, pulmonary hypertension is characterized by World Health Organization (WHO) groupings. In some implementations, pulmonary hypertension is WHO Group 1 pulmonary hypertension or pulmonary arterial hypertension (PAH), WHO Group 2 pulmonary hypertension, WHO Group 3 pulmonary hypertension, WHO Group 4 pulmonary hypertension, and WHO Group 5 pulmonary hypertension. In some implementations, pulmonary hypertension is characterized by the World Health Organization (WHO) classification system. In some implementations, pulmonary hypertension is characterized by the WHO functional classification based on cardiac function. In some implementations, pulmonary hypertension includes WHO Group I pulmonary hypertension or pulmonary arterial hypertension (PAH), WHO Group II pulmonary hypertension, WHO Group III pulmonary hypertension, and WHO Group IV pulmonary hypertension.

[0152] In some embodiments, pulmonary hypertension is associated with pulmonary capillary angioma. In some embodiments, the disease is heart failure or right ventricular failure. In one aspect, the present invention provides a method of treating a subject with pulmonary arterial hypertension (PAH) comprising administering an integrin α5β1 inhibitor.

[0153] Pulmonary hypertension (PAH).

[0154] In one aspect, the present invention provides a method for treating pulmonary arterial hypertension (PAH), comprising administering an integrin α5β1 inhibitor (e.g., small molecule compounds and antibodies disclosed herein). PAH is characterized by a progressive increase in pulmonary vascular resistance, leading to right ventricular overload and ultimately heart failure. PAH causes progressive obstruction and decreased compliance of the pulmonary artery (PA), leading to right ventricular (RV) failure and premature death. Like cancer cells, PA smooth muscle cells (PASMCs) and endothelial cells (PAECs) exhibit expanded proliferation and anti-apoptosis as a response to increased PA stiffness caused by extracellular matrix (ECM) remodeling. Integrin signaling promotes PAH-PASMC and PAH-PAEC proliferation and anti-apoptosis, causing PA vascular remodeling, while simultaneously causing adaptive hypertrophy and fibrosis of the RV, leading to RV failure via PAH. The present invention is based in part on the finding that α5β1 integrin inhibition can reverse PA vascular remodeling and prevent RV dysfunction in PAH.

[0155] PAH is a chronic condition involving all layers of the pulmonary vessels. In PAH, vasoconstriction, structural changes in the pulmonary vessel walls (vascular remodeling), and thrombosis contribute to increased pulmonary vascular resistance. Structural and functional changes in the endothelium cause endothelial dysfunction. Increased vasoconstrictive factors (e.g., endothelin) and decreased vasodilatory capacity (e.g., less prostacyclin) contribute to both vasoconstriction and increased pulmonary vascular resistance. Current therapies targeting vasoconstriction may slow the progression of PAH or provide limited clinical improvement, but they have not been shown to significantly reduce overall PAH morbidity and mortality. These treatments do not affect the underlying structural changes in the pulmonary vessels—vascular remodeling.

[0156] Vascular remodeling in PAH is characterized by proliferative and obstructive changes involving many cell types, including endothelial cells, smooth muscle cells, and fibroblasts. Vascular remodeling itself can manifest as, for example, thickening of the pulmonary vascular media due to smooth muscle cell proliferation and hypertrophy, formation of neointima by smooth muscle cells and / or myofibroblasts, and / or formation of plexiform lesions composed of localized proliferation of endothelial cells, smooth muscle cells, lymphocytes, and mast cells. Vascular remodeling causes luminal obstruction, leading to pulmonary hypertension. Therapies addressing the proliferative aspects of PAH are needed.

[0157] In some implementations, pulmonary hypertension is pulmonary arterial hypertension (PVH). In some variants, PAH is idiopathic PAH. In some variants, PAH is familial PAH. In some variants, PAH is associated with persistent pulmonary hypertension in newborns. In some variants, PAH is associated with pulmonary venous occlusive disease.

[0158] In some implementations, pulmonary hypertension is associated with lung disease. In some implementations, the lung disease is idiopathic pulmonary fibrosis (IPF) or interstitial pneumonia (IIP). IPF is a type of idiopathic interstitial pneumonia (IIP), which in turn is a type of interstitial lung disease (also known as diffuse parenchymal lung disease (DPLD)). Interstitial lung disease involves alveolar epithelium, pulmonary capillary endothelium, basement membrane, perivascular and perilymphatic tissue. Other forms of idiopathic interstitial pneumonia include nonspecific interstitial pneumonia (NSIP), desquamative interstitial pneumonia (DIP), and acute interstitial pneumonia (AIP). Examples of known causes of interstitial lung disease include sarcoidosis, hypersensitivity pneumonitis, pulmonary Langerhans cell histiocytosis, asbestosis, and collagen vascular diseases such as scleroderma and rheumatoid arthritis.

[0159] Pulmonary fibrosis is the formation or appearance of excess fibrous connective tissue in the lungs.

[0160] Heart failure (HF).

[0161] In one aspect, the present invention provides a method for treating heart failure (HF), comprising administering an integrin α5β1 inhibitor (e.g., small molecule compounds and antibodies disclosed herein). Heart failure refers to any condition characterized by the heart's inability to pump an adequate supply of blood to the body. It is a physiological state in which cardiac output is insufficient to meet the body's needs, or only meets the body's needs under higher filling pressures. HF has a variety of underlying causes, including myocardial infarction, coronary artery disease, valvular disease, hypertension, and myocarditis. Chronic heart failure is associated with changes in neurohormonal activation and autonomic control. While these compensatory neurohormonal mechanisms provide valuable support to the heart under normal physiological conditions, they also play a fundamental role in the development and subsequent progression of HF.

[0162] For example, one of the body's main compensatory mechanisms for reduced blood flow in heart failure (HF) is to increase the amount of salt and water retained by the kidneys. Retaining salt and water, rather than excreting them through urine, increases blood volume in the bloodstream and helps maintain blood pressure. However, the larger blood volume also causes the heart muscle (especially the ventricles) to enlarge. As the heart chambers enlarge, the walls become thinner and the heart's contractions weaken, leading to a spiral decline in cardiac function. Another compensatory mechanism is vasoconstriction in the arterial system, which raises blood pressure to help maintain adequate perfusion, thereby increasing the workload the heart must pump.

[0163] In heart failure with a low ejection fraction (EF), cardiac hypertension arises from the high pressure required by the body to maintain adequate peripheral perfusion. However, as the heart weakens due to this high pressure, the condition worsens. Left atrial pressure can exceed 25 mmHg. At this stage, blood fluid leaks out of the pulmonary circulation system or from the pulmonary capillaries into the interstitial spaces of the lungs and into the alveoli, causing pulmonary congestion. If left untreated, this can lead to acute pulmonary edema syndrome and death.

[0164] Standardized System of Care (SoC) and Combination Therapy As described herein, methods of treating diseases associated with enhanced integrin α5β1 expression or activity (including administration of an integrin α5β1 inhibitor) may include combination therapy, wherein an integrin α5β1 inhibitor is administered in combination with one or more drugs approved for the treatment of PH, PAH, heart failure, or right ventricular failure to a patient in need of treatment.

[0165] Currently approved medications in the United States or the European Union (EU) for the treatment of PH, PAH, heart failure, and right ventricular failure include orally administered PDE-5 inhibitors: sildenafil (Revatio) and tadalafil (Adeirca); dual endothelin-1A receptor antagonists (ERAs): bosentan (Tracleer) and ambrisentan (Letairis in the US; Volibris internationally). Patients with more advanced disease are typically treated with prostacyclin or prostacyclin analogues, such as iloprost (Ventavis) or treprostinil (Tyvaso) administered by inhalation multiple times daily; epoprostenol (Flolan / Veletri) or treprostinil (Remodulin) administered by continuous intravenous infusion; or treprostinil administered by continuous subcutaneous infusion. Intravenous sildenafil is approved for use in patients currently prescribed for treatment but temporarily unable to receive oral sildenafil. Inhaled nitric oxide (INOmax) is approved for neonatal forms of persistent pulmonary hypertension (PPHN) in newborns. Therefore, according to the invention, combination therapy of any of these drugs with an integrin a5b1 inhibitor is suitable for the treatment of PAH or the conditions disclosed herein.

[0166] In some implementations, the second therapy is selected from the following: anticoagulants, diuretics, digitalis glycosides, calcium channel blockers, endothelin receptor antagonists, phosphodiesterase 5 (PDE5) inhibitors, prostaglandins, prostaglandin receptor agonists, soluble guanylate cyclase stimulating factors, and / or surgery.

[0167] In some implementations, the second therapy is oxygen, warfarin, furosemide, bumetanide, bendroflumethiazide, metoprazone, spironolactone, amiloride, digoxin, nifedipine, diltiazem, nicardipine, amlodipine, ambrisentan, bosentan, macitentan, sildenafil, tadalafil, eprostol, iloprost, treprostacycline, riociguat, selexipag, surgery, pulmonary endocardiography, and / or atrial septalostomy. In some implementations, the second therapy is macitentan and / or tadalafil. Flolan (a prostacyclin analogue) is an approved treatment for PAH, but its administration (intravenous) is extremely cumbersome and inconvenient, and it presents unique safety concerns. Therefore, Flolan is typically reserved for patients with severe functional status or rapidly progressive PAH. Patients must reconstitute the medication multiple times daily under sterile conditions. The medication is available as a lyophilized formulation, which must be dissolved in an alkaline buffer. Due to its short half-life (3-5 minutes) and stability (8 hours at room temperature), Floran must be kept refrigerated while being administered continuously via a central venous catheter and a portable pump worn in a lap bag (CADD pump, Smith's MedicalMD, St. Paul, Minn.). In 2008, the FDA also approved a novel continuous intravenous formulation of eprostol that is stable at room temperature for up to 24 hours after dilution and can be stored at refrigerated temperatures for up to 5 days before use (GeneraMedix Inc., Liberty Corner, NJ). In 2009, GeneraMedix Inc. sold this formulation to Actelion, which began marketing the drug in April 2010 (under the brand name Veletri). In late 2010, the Veletri label was expanded to allow the preparation of formulations that can be stored at refrigerated temperatures for up to 7 days or at room temperature for up to 48 hours before use. Therefore, in one embodiment of the invention, an integrin a5b1 inhibitor is administered in combination with any approved form of eprostol to treat PAH.

[0168] Remodulin (a subcutaneously infused form of a prostacyclin analogue) is generally not used as initial therapy due to its cost, delivery route, and limited efficacy. In 2004, the FDA and Health Canada approved intravenous remodulin formulations for patients with PAH class II-IV who cannot tolerate the subcutaneous form. In early 2006, the FDA expanded the remodulin label to include patients requiring replacement of Flolan. In 2009, United Therapeutics' inhaled treprostacyclin formulation (Tyvaso) received FDA approval. Therefore, in one embodiment of the invention, an integrin a5b1 inhibitor is administered in combination with treprostacyclin to treat PAH.

[0169] Ventavis (iloprost) (a prostacyclin analog administered via inhalation) is also marketed as Ilomedine in intravenous formulation in several EU member states. In the EU, the labeling of inhaled iloprost is limited to patients with idiopathic PAH and functional class III symptoms. In contrast, the labeling in the US is broader: patients with PAH (regardless of etiology) and class III or IV symptoms. It requires 6 to 9 administrations per day. Therefore, in one embodiment of the invention, an integrin a5b1 inhibitor is combined with any approved form of iloprost to treat PAH.

[0170] In 2001, non-selective ERA Tracleer (bosentan) became the first oral treatment for PAH and was only available in the United States through a special centralized access program due to its significant risks of (reversible) liver damage, teratogenicity, testicular atrophy, and male infertility. Tracleer treatment consisted of an initial dose of 62.5 mg twice daily for 4 weeks, followed by a maintenance dose of 125 mg twice daily. Tracleer was initially designated for patients with PAH and moderate to severe functional status (WHO Class III or IV). In 2008 (EU) and 2009 (US), the label was expanded to include patients with mild symptoms (functional Class II). Therefore, in one embodiment of the invention, an integrin a5b1 inhibitor is administered in combination with any approved form of bosentan to treat PAH.

[0171] Ambrisentan is an oral selective ERA receptor antagonist, marketed in the United States by Gilead Sciences (Letairis) and in other regions by GlaxoSmithKline (Volibris), for once-daily treatment of patients with WHO Category II or III symptoms to improve motor function and delay clinical deterioration. Like bosentan, ambrisentan's side effects include teratogenicity, testicular damage, decreased male fertility, and anemia. Therefore, in one embodiment of the invention, an integrin a5b1 inhibitor is administered in combination with any approved form of ambrisentan to treat PAH.

[0172] Revatio (sildenafil), an oral PDE-5 inhibitor administered at a dose of 20 mg three times daily, is approved in the United States for the treatment of PAH (WHO Group I) to improve motor function and delay clinical deterioration, regardless of functional classification or etiology. The EU label is limited to improving motor function in patients with PAH that is idiopathic or associated with collagen vascular disease and functional Class III conditions. In 2009, the FDA approved an intravenous administration of Revatio (10 mg three times daily) for patients who cannot take the oral formulation. In May 2010, the EU approved Revatio as an oral suspension (composed of 20 mg tablets) for the treatment of pediatric patients aged 1 to 17 years with PAH. Therefore, in one embodiment of the invention, an integrin a5b1 inhibitor is administered in combination with any approved form of sildenafil to treat PAH.

[0173] In the United States, the once-daily oral PDE-5 inhibitor Adeirca (tadalafil) at a dose of 40 mg is designated for improving motor function in patients with PAH (WHO Group I), regardless of etiology or functional classification (label). The EU label is limited to patients with functional Group II and III conditions. Tadalafil has a long half-life (35 hours) in PAH patients (US label) and has also been shown to be beneficial in PAH patients when concomitant with bosentan.

[0174] Therefore, treatment of patients may involve the administration of at least one other active agent, i.e., an active agent other than an integrin a5b1 inhibitor. Other active agents may be, for example, vasodilators such as prostacyclin, eprostacyclin, and sildenafil; endothelin receptor antagonists such as bosentan; calcium channel blockers such as amlodipine, diltiazem, and nifedipine; anticoagulants such as warfarin; diuretics; prostaglandins (e.g., prostacyclin or PGI2); drugs used to treat diseases associated with overactivated or dysfunctional B cells, such as rituximab; and / or phosphodiesterase type V (PDE5) inhibitors.

[0175] When the methods of the present invention involve combination therapy (i.e., where a second agent (such as a vasodilator) is co-administered with an integrin a5b1 inhibitor), the agents may be administered separately, simultaneously, or at different times of the day, or may be administered as a single composition. Therefore, the present invention provides novel pharmaceutical formulations in which an integrin a5b1 inhibitor is combined with one of the active agents discussed above and unit dosage forms of these formulations.

[0176] In the combination therapy of the present invention, each agent may be administered in an "immediate-release" or "controlled-release" manner. When the other active agent is, for example, a vasodilator, any dosage form containing two active agents (i.e., an integrin a5b1 inhibitor and a vasodilator) may provide immediate or controlled release of the vasodilator and the integrin a5b1 inhibitor.

[0177] As a general example, a once-daily combination formulation of the present invention may contain about 1 mg to about 1000 mg of an integrin a5b1 inhibitor in a controlled-release (e.g., sustained-release) or immediate-release form, and an immediate-release or controlled-release form of sildenafil, wherein another active agent is present in an amount providing the integrin a5b1 inhibitor to sildenafil as specified above, or in the integrin a5b1 inhibitor to sildenafil weight ratio specified above. In other formulations of the present invention, two or more other active agents, which may or may not belong to the same class of drugs (e.g., vasodilators), may be present in combination with the integrin a5b1 inhibitor. In such cases, the effective amount of any or each of the individual other active agents present will generally be reduced compared to the amount required when using only a single additional agent.

[0178] As discussed above, other active agents may also be phosphodiesterase type V (PDEV) inhibitors administered in combination with an integrin α5b1 inhibitor or in combination with an integrin α5b1 inhibitor and a vasodilator. Examples of PDEV inhibitors include, but are not limited to, avanafil, sildenafil, tadalafil, zaprinast, dipyridamole, vardenafil, and acid adducts or other pharmaceutically acceptable salts thereof. Sildenafil is a prime example. In one exemplary embodiment, the integrin α5b1 inhibitor is co-administered with a PDEV inhibitor selected from avanafil, tadalafil, and sildenafil, and the daily dose of the integrin α5b1 inhibitor compound is as stated above for monotherapy.

[0179] In one embodiment, the vasodilator is selected from sildenafil, avanafil, tadalafil, zaplastase, dipyridamole, vardenafil, bosentan, and pharmaceutically acceptable salts thereof. As discussed above, other active agents may also be endothelin receptor antagonists, such as bosentan, sitaxentan, or ambrisentan, with bosentan being an illustrative active agent.

[0180] The pharmaceutical compositions of the present invention are pharmaceutical formulations containing an active agent, formulated in a manner compatible with a predetermined route of administration. Various routes are considered, including but not limited to oral, pulmonary, inhalation, sublingual, intranasal, parenteral, intradermal, transdermal, topical, mucosal, subcutaneous, intravenous, intramuscular, intraperitoneal, buccal, and rectal administration. As used herein, the term "parenteral" is intended to include subcutaneous, intravenous, and intramuscular injection.

[0181] Generally, the pharmaceutical formulations of the present invention are prepared in an immediate-release form suitable for oral administration and once-daily (QD) administration. Some formulations are suitable for intranasal administration.

[0182] Certain pharmaceutical formulations of the present invention comprise an integrin α5b1 inhibitor or a salt thereof and one or more pharmaceutically acceptable (approved for human use by a state or federal regulatory agency, or listed in the United States Pharmacopeia or the European Pharmacopoeia) excipients or carriers. As used herein, the term excipient or carrier broadly refers to a biologically inactive substance used in combination with an active agent in a formulation. Excipients may be used as, for example, solubilizers, stabilizers, diluents, inert carriers, preservatives, binders, disintegrants, coating agents, flavoring agents, or coloring agents. Preferably, at least one excipient is selected to impart one or more beneficial physical properties to the formulation, such as enhanced stability and / or solubility of the active agent. As described herein, integrin α5b1 inhibitors or salts thereof are exemplary active agents suitable for formulations of the present invention.

[0183] Examples of suitable excipients include certain inert proteins, such as albumin; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as aspartate, glutamate, lysine, arginine, glycine, and histidine; fatty acids and phospholipids, such as alkyl sulfonates and caprylates; surfactants, such as sodium dodecyl sulfate and polysorbate; nonionic surfactants, such as TWEEN®, PLURONICS®, or polyethylene glycol (PEG); carbohydrates, such as glucose, sucrose, mannose, maltose, trehalose, and dextrins, including cyclodextrins; polyols, such as mannitol and sorbitol; chelating agents, such as EDTA; and salt-forming ions, such as sodium.

[0184] Solutions or suspensions intended for delivery may include the following components: sterile diluents, such as water for injection, saline solution, non-volatile oils, polyethylene glycol, glycerol, propylene glycol, polysorbate, tocopheryl succinate (TPGS), or other synthetic solvents; antibacterial agents, such as benzyl alcohol or methylparaben; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates, or phosphates; and tonic modifiers, such as sodium chloride or dextrose. The pH may be adjusted with an acid or base (such as hydrochloric acid or sodium hydroxide). These formulations may be packaged in ampoules, disposable syringes, or multi-dose vials made of glass or plastic.

[0185] In some embodiments, the pharmaceutical formulations of the present invention contain a plurality of liposomes or microparticles comprising an integrin a5b1 inhibitor active agent. In various embodiments, the pharmaceutical formulation of the integrin a5b1 inhibitor is a powder comprising solid particles (e.g., liposomes or microparticles) suitable for administration by inhalation. The solid particles comprise an active agent, a carrier, optionally present surfactants, and optionally present other acceptors. The powder can be prepared by any convenient method. One example of a preparation method is spray-drying a solution containing the active agent (and other components) into a powder comprising a carrier compound. Another example is freeze-drying a solution comprising all components of the final powder.

[0186] Liposomes in formulations suitable for use in this invention are known in the art. For example, suitable liposomes comprise cholesterol, 1,2-distearyl-sn-glycero-3-phosphocholine (DSPC), and PEG-DSPE in a weight ratio of about 5:10:1. In some embodiments, the liposome formulation contains about 0.1-25% (e.g., 0.1%, 1%, 5%, 10%, or 20% (w / w)) of phospholipids, such as dipalmitoylphosphatidylcholine (DPPC) and 1,2-distearyl-sn-glycero-3-phosphocholine (DSPC). In some embodiments, the liposome formulation contains about 0.5-20% (e.g., 1%, 5%, or 10% (w / w)) of a hydrophilic polymer, such as polyvinylpyrrolidone (PVP). In some embodiments, the liposome formulation contains about 10-35% of an amino acid, such as L-leucine.

[0187] The microparticles in formulations suitable for use in this invention are known in the art. For example, the microparticles are formed from one or more hydrophilic polymers, such as polyvinylpyrrolidone (e.g., PVP-10), polyvinyl alcohol (e.g., PVA-30), polyvinyl acetate, or poloxamer (e.g., poloxamer-188). In some embodiments, the microparticle formulation comprises about 70-85% by weight of polyvinyl alcohol (e.g., PVA-30), about 5-15% PVP (e.g., PVP-10), 1-5% poloxamer (e.g., poloxamer-188), 0-10% L-leucine, and about 0.5-10% of an integrin a5b1 inhibitor compound (e.g., 5%). In some embodiments, the formulation is suitable for administration via the respiratory tract.

[0188] Pharmaceutical formulations of integrin α5b1 inhibitors suitable for use in the methods of this invention can be prepared as liquids or solids, such as orally administered powders, tablets, pills, or capsules. Liquid formulations of this invention can be in the form of suspensions, solutions, or emulsions in oily or aqueous media, and may contain formulations such as suspending agents, stabilizers, and / or dispersants. In one embodiment, the formulation is an aqueous solution. In another embodiment, the final formulation is lyophilized. In some embodiments, the integrin α5b1 inhibitor is formulated for inhalation.

[0189] In various embodiments, the formulations of the present invention comprise an integrin a5b1 inhibitor at a concentration of 0.25% to 100% by weight, or 0.25% to 50% by weight, or 0.8% to 25% by weight, or 1% to 10% by weight, or 1.5% to 5% by weight. In some embodiments, the integrin a5b1 inhibitor compound is formulated at a concentration of about 0.5% to about 5% by weight. In some embodiments, the integrin a5b1 inhibitor compound is formulated at a concentration of about 0.25% to about 10% by weight.

[0190] The present invention also provides a pharmaceutical package or kit comprising one or more containers filled with a solid or liquid formulation of an integrin a5b1 inhibitor. In one particular embodiment, the formulation is a powder formulation of an integrin a5b1 inhibitor. In various embodiments, the integrin a5b1 inhibitor is formulated at a concentration of at least about 0.5% by weight and the formulation is suitable for delivery to a human via inhalation.

[0191] This invention also provides formulations of integrin a5b1 inhibitors for use in the manufacture of PAHs or the conditions disclosed herein for the treatment of subjects in need. Generally, the pharmaceutical formulations are sterile.

[0192] Generally, dosage forms (e.g., inhalable formulations) allow the compounds of the present invention (e.g., integrin α5b1 inhibitors) to be released continuously (i.e., gradually) into the patient's body over an extended period, typically keeping blood drug levels substantially constant over a period of about 4 to about 12 hours, and typically over a period of about 6 to about 10 hours. In a particularly preferred embodiment, after nasal administration of a dosage form containing the compounds of the present invention (e.g., integrin α5b1 inhibitors), blood drug levels increase very slowly, such that peak blood drug levels are not reached until at least 4-6 hours have elapsed, and the rate of increase in blood drug levels is approximately linear. Additionally, in a preferred embodiment, blood drug levels also decrease slowly at the end of the sustained release period.

[0193] Although the pharmaceutical compositions of the present invention are preferably formulated for inhalation, such as in the form of a solution in saline, a dry powder, or an aerosol, other modes of administration are also suitable. For example, they can be administered sublingually, orally, non-intestinally, percutaneously, via an implanted depot, or via a mucosa (e.g., rectal or vaginal, preferably using suppositories containing excipients other than the active agent, such as suppository wax). Mucosal administration also includes transurethral administration.

[0194] Depending on the intended administration method, the pharmaceutical formulation may be solid, semi-solid, or liquid, such as tablets, capsules, granules, liquids, suspensions, emulsions, suppositories, granules, pellets, beads, powders, etc., preferably in a unit dosage form suitable for a precise single dose. Suitable pharmaceutical compositions and dosage forms can be prepared using conventional methods known in the field of pharmaceutical formulation and described in relevant textbooks and literature, such as Remington: The Science and Practice of Pharmacy (Easton, Pa.: Mack Publishing Co., 1995). For compounds with oral activity, oral dosage forms are generally preferred and include tablets, capsules, granules, solutions, suspensions, and syrups, and may also contain multiple granules, beads, powders, or pellets, which may or may not be encapsulated. Preferred oral dosage forms are tablets and capsules.

[0195] In embodiments, it is particularly advantageous to formulate the compositions of the invention into unit dosage forms for ease of administration and dosage uniformity. As used herein, the term "unit dosage form" refers to a physical, discontinuous unit suitable as a unit dose for use on an individual to be treated. That is, the composition is formulated into discontinuous dose units, each containing a predetermined "unit dosage form" quantity of the active agent and the desired drug carrier, the quantity of which is calculated to produce the desired therapeutic effect. The specifications of the unit dosage forms of the invention are based on the unique characteristics of the active agent to be delivered. The dosage can be further determined by referring to the commonly used dosages and methods of administration of the components. It should be noted that in some cases, a combination of two or more individual dose units provides a therapeutically effective amount of the active agent; for example, two tablets or capsules taken together can provide a therapeutically effective dose of an integrin a5b1 inhibitor, such that the unit dosage form of each tablet or capsule is approximately 50% of the therapeutically effective amount.

[0196] Tablets can be manufactured using standard tablet processing procedures and equipment. Direct tableting and granulation techniques are preferred. In addition to active agents, tablets typically contain pharmaceutically acceptable inactive carrier materials, such as binders, lubricants, disintegrants, fillers, stabilizers, surfactants, colorants, etc.

[0197] Capsules are another oral dosage form of those compounds of the present invention (e.g., integrin a5b1 inhibitors) that have oral activity, in which case the composition containing the active agent can be encapsulated in liquid or solid form (including microparticles, such as granules, beads, powders, or pellets). Suitable capsules can be hard or soft capsules and are generally made of gelatin, starch, or cellulose materials, with gelatin capsules being preferred. Two-stage hard gelatin capsules are preferably sealed, such as with gelatin tape. See, for example, Remington: The Science and Practice of Pharmacy, cited earlier herein, which describes materials and methods for preparing encapsulated pharmaceutical products.

[0198] If desired, the formulation may be prepared into an oral dosage form (whether tablets, capsules, granules or microparticles) to provide controlled release of the compounds of the present invention (e.g., integrin a5b1 inhibitors), and in a preferred embodiment, the formulation of the present invention is a controlled-release oral dosage form.

[0199] Generally, as those skilled in the art will understand, sustained-release dosage forms are formulated by dispersing the active agent in a matrix of a gradually hydrolyzable material (such as a hydrophilic polymer) or by coating a solid dosage form containing the drug with such a material. Suitable hydrophilic polymers for providing sustained-release coatings or matrices include, for example: cellulose polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, methylcellulose, ethyl cellulose, cellulose acetate, and sodium carboxymethyl cellulose; acrylic polymers and copolymers, preferably formed from acrylic acid, methacrylic acid, alkyl acrylates, alkyl methacrylates, etc., such as copolymers of acrylic acid, methacrylic acid, methyl acrylate, ethyl acrylate, methyl methacrylate, and / or ethyl methacrylate; and vinyl polymers and copolymers such as polyvinylpyrrolidone, polyvinyl acetate, and ethylene-vinyl acetate copolymers.

[0200] Non-entericidal formulations according to the invention include aqueous and non-aqueous sterile solutions, suspensions, and emulsions. Injectable aqueous solutions contain the active agent in a water-soluble form. Examples of non-aqueous solvents or media include fatty oils such as olive oil and corn oil; synthetic fatty acid esters such as ethyl oleate or triglycerides; low molecular weight alcohols such as propylene glycol; and synthetic hydrophilic polymers such as polyethylene glycol, liposomes, etc. Non-entericidal formulations may also contain adjuvants such as solubilizers, preservatives, wetting agents, emulsifiers, dispersants, and stabilizers, and aqueous suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, and polydextrose. Injectable formulations are sterilized by incorporating a sterilizing agent, filtering via a bacterial trap filter, irradiation, or heating. They can also be prepared using sterile injectable media. The active agent may also be in a dried (e.g., lyophilized) form, which can be rehydrated with a suitable media before being administered by injection.

[0201] Conventional transdermal drug delivery systems can also be used to administer the active agent through the skin, wherein the active agent is contained within a laminated structure that acts as a drug delivery device adhered to the skin. In such a structure, the drug composition is contained in a layer or "reservoir" beneath an upper backing layer. The laminated structure may contain a single reservoir, or it may contain multiple reservoirs. In one embodiment, the reservoir contains a polymer matrix of a pharmaceutically acceptable adhesive material for attaching the system to the skin during drug delivery. Alternatively, the drug-containing reservoir and the skin-contact adhesive layer exist as separate layers, wherein the adhesive layer beneath the reservoir may in this case be a polymer matrix as described above, or it may be a liquid or hydrogel reservoir, or some other form. The transdermal drug delivery system may additionally contain a skin penetration enhancer.

[0202] In addition to the formulations described above, the active agent can also be formulated as a reservoir-type formulation for controlled release of the active agent, preferably a sustained release over an extended period of time. These sustained-release formulations are typically administered via implantation (e.g., subcutaneous or intramuscular administration or intramuscular injection).

[0203] Some compounds or active agents of the present invention can further form salts. All such forms are also contemplated within the scope of protection claimed by the present invention. The compounds of the present invention can also be prepared into esters, such as pharmaceutically acceptable esters. For example, the carboxylic acid functional group in the compound can be converted into its corresponding ester, such as methyl ester, ethyl ester, or other esters. In addition, the alcohol group in the compound can be converted into its corresponding ester, such as acetate, propionate, or other esters.

[0204] Some compounds of the present invention can also be prepared as prodrugs, such as pharmaceutically acceptable prodrugs. The terms "prodrug" and "prodrug" are used interchangeably herein and refer to any compound that releases an active parent drug in vivo. Since prodrugs are known to enhance a variety of desired qualities of a drug (e.g., solubility, bioavailability, manufacturing, etc.), the compounds of the present invention can be delivered in prodrug form. Therefore, the present invention is intended to cover prodrugs of the compounds claimed herein, methods of their delivery, and compositions containing them. "Prodrug" is intended to include any covalently bonded carrier that releases the active parent drug of the present invention in vivo when such a prodrug is administered to a subject. Prodrugs of the present invention are prepared by modifying functional groups present in the compound such that the modification cleaves to the parent compound in conventional operation or in vivo. Prodrugs include compounds of the present invention wherein a hydroxyl, amino, sulfhydryl, carboxyl, or carbonyl group is bonded to any group that can be cleaved in vivo, forming a free hydroxyl, free amino, free sulfhydryl, free carboxyl, or free carbonyl group, respectively.

[0205] Examples of prodrugs include (but are not limited to) esters (e.g., acetates, dialkylaminoacetic acids, formates, phosphates, sulfates, and benzoate derivatives) and carbamates (e.g., N,N-dimethylaminocarbonyl) of the compounds of the present invention and their analogues; esters (e.g., ethyl esters, morpholinoethanol esters) of the carboxyl functional group; N-acyl derivatives (e.g., N-acetyl), N-Mannichbase, Schiffbase, and enamine ketones of the amino functional group; and oximes, acetals, ketals, and enol esters of the ketone and aldehyde functional groups, see Bundegaard, H., Design of Prodrugs, pp. 1-92, Elesevier, New York-Oxford (1985).

[0206] The dosing regimen for a compound is selected based on a number of factors, including the patient's type, species, age, weight, sex, and medical condition; the severity of the condition to be treated; the route of administration; the patient's renal or hepatic function; and the specific compound or its salt used. A skilled general practitioner or veterinarian can readily determine and specify the effective amount of drug required to prevent, resist, or halt the progression of the condition.

[0207] In some embodiments, the composition is suitable for inhalation. In one embodiment, the composition is an inhalable formulation for treating PAH or the conditions disclosed herein.

[0208] In another aspect, the present invention provides a pharmaceutical composition comprising an integrin a5b1 inhibitor and a plurality of particles, wherein the plurality of particles are a plurality of liposomes comprising 1,2-distearate-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)]] (PEG-DSPE) or a plurality of microparticles comprising a hydrophilic polymer. In one embodiment, the composition is suitable for inhalation; in one embodiment, the composition is an inhalable formulation for treating PAH or the conditions disclosed herein.

[0209] Pharmaceutical Composition The pharmaceutical compositions described herein can be administered in a variety of different ways. Examples include administration via oral, intranasal, rectal, topical, intraperitoneal, intravenous, intramuscular, subcutaneous, subdermal, percutaneous, intrathecal, and intracranial methods, wherein the pharmaceutical composition comprises a peptide or polymer (preferably a peptide) according to the invention and contains a pharmaceutically acceptable carrier. When administered orally, the active ingredient may be administered in solid dosage forms (e.g., capsules, tablets, and powders) or in liquid dosage forms (e.g., elixirs, syrups, and suspensions).

[0210] The pharmaceutical compositions according to the invention comprise at least one pharmaceutically acceptable carrier, diluent, or excipient. Examples of suitable carriers include keyhole limpet haemocyanin (KLH), serum albumin (e.g., BSA or RSA), and ovalbumin. In some embodiments, the suitable carrier is a solution, such as saline. Examples of excipients that can be incorporated into tablets, capsules, etc., include: binders such as gum arabic, gum arabic, corn starch, or gelatin; excipients such as microcrystalline cellulose; disintegrants such as corn starch, pregelatinized starch, alginic acid, etc.; lubricants such as magnesium stearate; sweeteners such as sucrose, lactose, or saccharin; and flavoring agents such as peppermint, wintergreen oil, or cherry. When the unit dosage form is a capsule, it may also contain a liquid carrier, such as fatty oil, in addition to the materials of the types described above. Various other materials may be present in coated form or otherwise modify the physical form of the dosage unit. For example, tablets may be coated with shellac, sugar, or both. The syrup or elixir may contain an active compound, sucrose as a sweetener, methylparaben and propylparaben as preservatives, dyes, and flavorings such as cherry or orange flavorings. The pharmaceutical compositions according to the invention are preferably suitable for human use.

[0211] The sterile composition for injection can be formulated according to conventional pharmaceutical practice as follows: the integrin a5b1 inhibitor of the present invention is dissolved or suspended in an injectable medium, such as water or naturally occurring vegetable oils, such as sesame oil, coconut oil, peanut oil, cottonseed oil, etc., or a synthetic fatty acid medium, such as ethyl oleate, etc. Buffers, preservatives, antioxidants, etc., may also be incorporated.

[0212] Topical compositions may also be formulated according to conventional pharmaceutical practice. As used herein, “topical application” means application to a body surface, such as skin or mucous membranes, for the local treatment of symptoms caused by microbial or parasitic infections. Examples of formulations suitable for topical application include (but are not limited to) creams, gels, ointments, lotions, foams, suspensions, sprays, aerosols, and powdered aerosols. Topical agents may be epidermal agents, meaning they are applied directly to the skin. Topical agents may also be inhaled agents, such as those applied to the mucous epithelium of the respiratory tract, or to tissue surfaces other than the skin, such as eye drops applied to the conjunctiva, or ear drops placed in the ear. Pharmaceutical compositions formulated for topical application preferably contain at least one pharmaceutical excipient suitable for topical application, such as emulsifiers, diluents, humectants, preservatives, pH adjusters, and / or water.

[0213] Exemplary implementation plan The following examples are illustrated to aid in understanding the invention and should not be construed as limiting the invention described and claimed herein. Such variations of the invention include replacing all currently known or later-developed equivalents, which will be within the purview of those skilled in the art, and variations in formulation or experimental design will be considered to fall within the scope of the invention incorporated herein.

[0214] Some of the following examples relate to methods for treating or preventing diseases that respond to α5β1 integrin inhibitors by administering a compound of formula (I) to a person who requires such treatment. Example

[0215] Certain compounds contained in the compositions of the present invention may exist in specific geometric or stereoisomeric forms. Furthermore, the polymers of the present invention may also be optically active. The present invention contemplates all such compounds falling within the scope of the invention, including cis and trans isomers. R -enantiomers and S - Enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures thereof. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are intended to be included in this invention.

[0216] For example, if a specific enantiomer of the compound of the present invention is required, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary agent, wherein the resulting mixture of diastereomers is separated and the auxiliary groups are cleaved to provide the desired enantiomer in pure form. Alternatively, in the case where the molecule contains a basic functional group such as an amino group or an acidic functional group such as a carboxyl group, a diastereomeric salt is formed with a suitable optically active acid or base, and the resulting diastereomer is then resolved by stepwise crystallization or chromatographic means known in the art, and the pure enantiomer is subsequently recovered. Example

[0217] abbreviation Abbreviated chemical names 9-BBN 9-boronabicyclo[3.3.1]nonane dimer Acetyl group ACN Acetonitrile APhosPdG3(4-(N,N-dimethylamino)phenyl)di-tert-butylphosphine,[4-(di-tert-butylphosphino)-N,N-dimethylaniline-2-(2′-aminobiphenyl)]palladium(II)methanesulfonate Boc tert-butoxycarbonyl Bu Butyl CMBP 4-Chloro-4′-maleimide benzophenone cod cyclooctadiene Cp cyclopentadienyl dba dibenzylidene acetone DCM dichloromethane DIAD (Diisopropyl Azodicarbonate) DIBAL (Diisobutylaluminum) DIEA N,N-Diisopropylethylamine DMF (dimethylformamide) DMSO (dimethyl sulfoxide) dppf 1,1-bis(diphenylphosphine)ferrocene dtbpy 4,4′-di-tert-butyl-2,2′-bipyridine Et Ethyl HATU (Hexafluorophosphate, benzotriazole, tetramethylurea) HMDS (Hexamethyldisilazane) Hz Hertz iPr isopropyl LDA Lithium diisopropylamide m-CPBA (m-chloroperoxybenzoic acid) Me methyl MOM (methoxymethyl) MTBE (methyl tert-butyl ether) NBS N-bromosuccinimide NMR (Nuclear Magnetic Resonance) Ph phenyl Pin-Pinol Piv 2,2-Dimethylacetyl preparative high-performance liquid chromatography (prep-HPLC) SFC Supercritical Fluid Chromatography TBAI Tetrabutylammonium Iodide t-Bu tert-butyl TEA Triethylamine Tf trifluoromethanesulfonyl TFA (trifluoroacetic acid) THF Tetrahydrofuran TLC (Thin Layer Chromatography) TMEDA Tetramethylethylenediamine TMS (trimethylsilyl) Ts p-Toluenesulfonyl group.

[0218] Analytical methods, materials and instruments Unless otherwise noted, reagents and solvents shall be used in the form received from commercial suppliers.

[0219] Left middle body Preparation of (1S,3S)-N-methyl-3-(4-(5,6,7,8-tetrahydro-1,8-naphthidin-2-yl)butoxy)cyclopentan-1-amine Step 1: (1S,3S)-3-(methylamino)cyclopentan-1-ol A solution of 1 / 3 of ((1S,3S)-3-hydroxycyclopentyl)carbamate (50 g, 248.43 mmol, 1 equivalent) was added dropwise to a solution of LiAlH4 (18.86 g, 496.86 mmol, 2 equivalents) in THF (500 mL) under N2 at 20 °C, and the solution was stirred at 80 °C. Then, a solution of 2 / 3 of ((1S,3S)-3-hydroxycyclopentyl)carbamate was slowly added to the solution at 80 °C under N2, and the solution was stirred at 80 °C for 2 hours. TLC (petroleum ether / ethyl acetate = 1 / 1) indicated that the starting material was completely exhausted and a new spot was formed. The reaction mixture was quenched dropwise by slow addition of H2O (20 mL), NaOH aqueous solution (15%, 20 mL), and H2O (60 mL) at 10 °C under N2. The mixture was dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to obtain a residue. (1S,3S)-3-(methylamino)cyclopentan-1-ol (28.61 g, crude) was obtained as a yellow oil. The residue was used directly without further purification.

[0220] Step 2: ((1S,3S)-3-hydroxycyclopentyl)(methyl)carbamate tert-butyl ester A solution of (1S,3S)-3-(methylamino)cyclopentan-1-ol (56 g, 486.23 mmol, 1 equivalent) in THF (600 mL) was added to NaHCO3 (81.69 g, 972.45 mmol, 37.82 mL, 2 equivalents) in H2O (90 mL) and (Boc)2O (137.95 g, 632.09 mmol, 145.21 mL, 1.3 equivalents), and the solution was stirred at 25 °C for 16 hours. TLC (petroleum ether / ethyl acetate = 1 / 1, R f= 0.4) indicates that the starting material has been completely exhausted and a new spot has formed. H2O (200 mL) was added to the reactants and extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). Tert-butyl ((1S,3S)-3-hydroxycyclopentyl)(methyl)carbamate (82.16% yield) was given as a yellow oil. 1 ¹H NMR (400 MHz, chloroform-d) δ=4.74 (brs, 1H), 4.39 (brs, 1H), 2.73 (s, 3H), 2.03–1.97 (m, 2H), 1.88–1.72 (m, 2H), 1.66–1.54 (m, 2H), 1.47 (s, 9H).

[0221] Step 3: ((1S,3S)-3-(4-bromobutoxy)cyclopentyl)(methyl)carbamate tert-butyl ester 1,4-Dibromobutane (170.49 g, 789.64 mmol, 95.25 mL, 2 equivalents) and TBAI (218.75 g, 592.23 mmol, 1.5 equivalents) were added to a solution of ((1S,3S)-3-hydroxycyclopentyl)(methyl)carbamate tert-butyl ester (85 g, 394.82 mmol, 1 equivalent) in toluene (1000 mL), NaOH (247.09 g, 6.18 mol, 500 mL, 15.65 equivalents), and the solution was stirred at 50 °C for 16 hours. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.6) indicates the presence of remaining starting material and the formation of a new spot. The reaction mixture was quenched by adding H2O (1000 mL) at 0 °C and extracted with ethyl acetate (300 mL × 3). The combined organic layers were washed with brine (300 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1). Tert-butyl ((1S,3S)-3-(4-bromobutoxy)cyclopentyl)(methyl)carbamate (28.92% yield) was given as a yellow oil. 1 ¹H NMR (400 MHz, chloroform-d) δ=4.61 (brs, ¹H), 3.91 (brd, J=2.9Hz,1H), 3.47-3.37 (m, 4H), 2.72 (s, 3H), 2.00-1.82 (m, 6H), 1.72-1.62 (m, 4H), 1.46 (s, 9H).

[0222] Step 4: ((1S,3S)-3-(but-3-en-1-yloxy)cyclopentyl)(methyl)carbamate tert-butyl t-BuOK (26.43 g, 235.52 mmol, 2.5 equivalents) was added to a solution of ((1S,3S)-3-(4-bromobutoxy)cyclopentyl)(methyl)carbamate (tert-butyl ester) (33 g, 94.21 mmol, 1 equivalent) in THF (400 mL) at 0 °C, and the mixture was stirred at 20 °C for 16 hours. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.61) indicates that the starting material was completely exhausted and a new spot was formed. The reaction mixture was quenched by adding H2O (100 mL) at 0 °C and extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1). Tert-butyl ((1S,3S)-3-(but-3-en-1-yloxy)cyclopentyl)(methyl)carbamate (90.63% yield) was given as a yellow oil. 1 ¹H NMR (400 MHz, chloroform-d) δ=5.82 (tdd, J =6.7, 10.3, 17.1 Hz, 1H), 5.14-4.99 (m, 2H), 4.61 (brs, 1H), 3.94-3.89 (m, 1H), 3.43-3.37 (m, 2H), 2.72 (s,3H), 2.32-2.27 (m, 2H), 1.99-1.84 (m, 4H), 1.69-1.61 (m, 2H), 1.46 (s, 9H).

[0223] Step 5: 7-(4-(((1S,3S)-3-((tert-butoxycarbonyl)(methyl)amino)-cyclobutyl)oxy)butyl)-3,4-dihydro-1,8-naphthidine-1(2H)-formate tert-butyl ester 9-BBN (0.5 M, 341.53 mL, 2 equivalents) was added to a solution of ((1S,3S)-3-(but-3-en-1-yloxy)cyclopentyl)(methyl)carbamate tert-butyl ester (23 g, 85.38 mmol, 1 equivalent) in THF (50 mL), and the mixture was stirred at 25 °C for 2 hours. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.53) indicates complete depletion of the starting material and the formation of a new peak. The above solution (34.87 g, 89.09 mmol, 1 equivalent) in tert-amyl alcohol (tAmylOH) (400 mL) was supplemented at 25 °C with tert-butyl 7-chloro-3,4-dihydro-1,8-naphthidine-1(2H)-carboxylate (21.55 g, 80.18 mmol, 0.9 equivalent), Aphos Pd G3 (5.66 g, 8.91 mmol, 0.1 equivalent), and Cs2CO3 (58.06 g, 178.18 mmol, 2 equivalent). The mixture was stirred at 80 °C under N2 for another 14 hours. LC-MS showed complete depletion of the starting material and the detection of a main peak with the desired mass. The reaction mixture was quenched at 0 °C with H2O (500 mL) and extracted with ethyl acetate (200 mL × 3). The combined organic layers were washed with brine (200 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). 7-(4-(((1S,3S)-3-((tert-butoxycarbonyl)(methyl)amino)-cyclopentyl)oxy)butyl)-3,4-dihydro-1,8-naphthidine-1(2H)-carboxylic acid tert-butyl ester (62.40% yield) was given as a red oil. 1 ¹H NMR (400 MHz, chloroform-d) δ=7.23 (brd, J =7.6Hz, 1H), 6.75 (d, J =7.6Hz, 1H), 3.90-3.77 (m, 1H), 3.70-3.66(m, 2H), 3.31 (t, J =6.6Hz, 2H), 2.65 (brs, 3H), 2.64 (s, 3H), 1.92-1.74 (m,6H), 1.73-1.50 (m, 8H), 1.45 (s, 9H), 1.38 (s, 9H).

[0224] Step 6: (1S,3S)-N-methyl-3-(4-(5,6,7,8-tetrahydro-1,8-naphthidin-2-yl)butoxy)cyclopentan-1-amine TFA (153.50 g, 1.35 mol, 100.00 mL, 24.22 equivalents) was added to a solution of tert-butyl 7-(4-(((1S,3S)-3-((tert-butoxycarbonyl)(methyl)amino)-cyclopentyl)oxy)butyl)-3,4-dihydro-1,8-naphthyl-1(2H)-carboxylate (28 g, 55.59 mmol, 1 equivalent) in DCM (300 mL), and the solution was stirred at 25 °C for 16 h. LCMS showed complete depletion of the starting material and detection of a main peak with the desired mass. The residue was adjusted to pH 7 with aqueous NaHCO3 solution, extracted with DCM (100 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (column: Welch Xtimate C18 250 × 100 mm × 10 μm; mobile phase: [water (NH3H2O ​​+ NH4HCO3)-ACN]; B%: 0%-50%, 35 min) to give (1S,3S)-N-methyl-3-(4-(5,6,7,8-tetrahydro-1,8-naphthid-2-yl)butoxy)cyclopentan-1-amine (54.54% yield) as a yellow oil.

[0225] LCMS :[M+1]=304.2;SFC:R t =2.070min; 1 H NMR (400 MHz, methanol-d4) δ=7.11 (d,J=7.4Hz, 1H), 6.35 (d, J=7.4Hz, 1H), 4.07-3.82 (m, 1H), 3.41-3.34 (m, 4H), 3.10 (quin, J=7.1 Hz, 1H), 2.68 (t, J=6.3Hz, 2H), 2.51 (t, J=7.6Hz, 2H), 2.32(s, 3H), 2.03-1.91 (m, 3H), 1.90-1.83 (m, 2H), 1.71-1.59 (m, 3H), 1.57-1.46(m, 3H), 1.38-1.28(m, 1H).

[0226] Preparation of rac-(1S,4R)-N,3,3-trimethyl-4-(4-(5,6,7,8-tetrahydro-1,8-naphthidin-2-yl)butoxy)cyclopentan-1-amine hydrochloride Step 1: 5-Bromo-2,2-dimethylcyclopentan-1-one A solution of Br2 (391.8 g, 2.45 mol, 126.31 mL, 1.1 equivalent) in DCM (500 mL) was added to a solution of 2,2-dimethylcyclopentan-1-one (250 g, 2.23 mol, 279.64 mL, 1 equivalent) in DCM (2000 mL) at 0 °C. The mixture was allowed to stir at 20 °C for 16 hours. The reaction was quenched by adding H2O (2000 mL) and saturated NaS2O3 was added at 25 °C until the solution became colorless. The pH of the solution was adjusted to pH=7 by adding saturated NaHCO3. The mixture was diluted with DCM (2000 mL) and extracted with DCM (1000 mL × 3). The organic solutions were combined, washed with brine (1000 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give 5-bromo-2,2-dimethylcyclopentan-1-one (350 g, crude substance) as a brown solid.

[0227] Step 2: 5,5-Dimethylcyclopent-2-en-1-one A mixture of 5-bromo-2,2-dimethylcyclopentan-1-one (350 g, 1.83 mol, 1 equivalent) and DBU (334.65 g, 2.20 mol, 331.34 mL, 1.2 equivalent) in MTBE (2000 mL) was degassed and purged three times with nitrogen. The mixture was allowed to be stirred at 25 °C under N2 atmosphere for 16 hours. The mixture was then distilled under vacuum (62 °C, -0.08 MPa / water pump). 5,5-Dimethylcyclopentan-2-en-1-one (110 g, 798.89 mmol, 44.81% yield via two steps) was given as a pale yellow oil.

[0228] Step 3: 4-(benzyl(methyl)amino)-2,2-dimethylcyclopentan-1-one BnMeNH (114.96 g, 948.68 mmol, 122.43 mL, 0.95 equivalents) was added dropwise to 5,5-dimethylcyclopentan-2-en-1-one (110 g, 998.61 mmol, 1 equivalent) at 25 °C, and the mixture was stirred at 50 °C for 2 hours. The reaction mixture was cooled to 0 °C and a solid was formed. The mixture was filtered and the filter cake was dried under vacuum. 4-(benzyl(methyl)amino)-2,2-dimethylcyclopentan-1-one (51.95% yield) was given as a yellow solid.

[0229] Step 4: (3,3-Dimethyl-4-oxocyclopentyl)(methyl)carbamate tert-butyl ester (Boc)₂O (84.91 g, 389.05 mmol, 89.38 mL, 1.5 equivalents) and 4-(benzyl(methyl)amino)-2,2-dimethylcyclopentan-1-one (60 g, 259.37, 1 equivalent) were added to a solution of Pd / C (5 g, 10% purity) in MeOH (1000 mL) at 25 °C under Ar conditions. The suspension was degassed under vacuum and purged three times with H₂. The mixture was allowed to be stirred at 25 °C under H₂ (30 psi) for 12 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1). Tert-butyl (3,3-dimethyl-4-oxocyclopentanyl)(methyl)carbamate (51.92% yield) was given as a pale yellow oil. 1 H NMR (400 MHz, CDCl3) δ=4.81 (brd, J =1.0Hz, 1H), 2.77 (s, 3H), 2.61-2.48 (m, 1H), 2.39-2.25 (m, 1H), 2.02-1.93 (m,1H), 1.89-1.77 (m, 1H), 1.47 (s, 9H), 1.13 (s, 3H), 1.06 (s, 3H).

[0230] Step 6: rac-((1S,4R)-4-hydroxy-3,3-dimethylcyclopentyl)(methyl)carbamate tert-butyl ester DIBAL-H (1 M, 538.69 mL, 1 equivalent) was added to a solution of (3,3-dimethyl-4-oxocyclopentyl)(methyl)carbamate tert-butyl ester (130 g, 538.69 mmol, 1 equivalent) in THF (1000 mL) under a N2 atmosphere at 0 °C. The reaction mixture was then heated to 20 °C and stirred at 20 °C under a N2 atmosphere for 16 h. The mixture was quenched by adding Na2SO4·10 H2O (100 g). The mixture was then dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1). rac-((1S,4R)-4-hydroxy-3,3-dimethylcyclopentyl)(methyl)carbamate tert-butyl ester (41.96% yield) was given as a colorless oil. 1H NMR (400 MHz, CDCl3) δ=4.72 (m, 1H), 3.83 (t, J =7.8 Hz, 1H), 2.73 (s, 3H), 2.06-1.96 (m,2H), 1.88 (brdd, J =7.7, 10.4Hz, 1H), 1.75 (dd, J =8.6, 12.9Hz, 1H), 1.46 (s,9H), 1.06 (s, 3H), 0.94 (s, 3H).

[0231] Final step: rac-(1S,4R)-N,3,3-trimethyl-4-(4-(5,6,7,8-tetrahydro-1,8-naphthidin-2-yl)butoxy)cyclopentan-1-amine hydrochloride By a method similar to that described above for the synthesis of rac-(1S,3S)-N-methyl-3-(4-(5,6,7,8-tetrahydro-1,8-naphthidin-2-yl)butoxy)cyclopentan-1-amine in steps 3-6, rac-((1S,4R)-4-hydroxy-3,3-dimethylcyclopentyl)(methyl)carbamate tert-butyl ester was converted to rac-(1S,4R)-N,3,3-trimethyl-4-(4-(5,6,7,8-tetrahydro-1,8-naphthidin-2-yl)butoxy)cyclopentan-1-amine hydrochloride. LCMS [M+1] = 332.2, 96.78% 1 H NMR (400MHz, CD3OD) δ=7.58 (d, J =7.3Hz, 1H), 6.60 (d, J =7.4Hz, 1H), 3.64 (brdd, J =6.8,8.5Hz, 1H), 3.57 (s, 5H), 2.82 (t, J =6.1 Hz, 2H), 2.73 (t, J =7.7Hz, 2H), 2.65(s, 3H), 2.13-1.93 (m, 5H), 1.82-1.73 (m, 2H), 1.62 (brdd, J =6.2, 8.7Hz, 2H), 1.45 (dd, J =8.8, 13.1 Hz, 1H), 1.11 (s, 3H), 0.94 (s, 3H).

[0232] Right-side intermediate Preparation of 1-bromo-3-fluoro-4-methyl-2-nitrobenzene Step 1: 3-Fluoro-4-methyl-2-nitroaniline Two reactions were carried out in parallel. Pd(dppf)Cl2 (3.89 g, 5.32 mmol, 0.05 equivalent) and K2CO3 (44.11 g, 319.13 mmol, 3 equivalent) were added to a mixture of 4-bromo-3-fluoro-2-nitroaniline (25 g, 106.38 mmol, 1 equivalent) and methylboronic acid (31.84 g, 531.89 mmol, 5 equivalent) in dioxane (250 mL) and H2O (62.5 mL) at 25 °C. The mixture was allowed to stir at 90 °C under a N2 atmosphere for 16 hours. The mixture was quenched by adding H2O (300 mL) and filtered. The filtrate was extracted with ethyl acetate (200 mL × 3). The organic solutions were combined, washed with brine (150 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1). 3-fluoro-4-methyl-2-nitroaniline was given as a yellow oil (55.25% yield). LCMS: R t =0.557min, [M+1]=171.1, 55.30%. 1 H NMR (400 MHz, CDCl3) δ=7.11 (t, J =8.1 Hz, 1H), 6.50 (dd, J =1.6, 8.6Hz, 1H), 2.19 (d, J =2.5Hz, 3H).

[0233] Step 2: 1-Bromo-3-fluoro-4-methyl-2-nitrobenzene Add 3-fluoro-4-methyl-2-nitroaniline to a solution of HBr (111.75 g, 455.77 mmol, 75.00 mL, 33% purity, 15.51 equivalents) in H2O (500 mL) and allow the mixture to stir at 70 °C for 30 minutes. Cool the mixture to -5 °C and slowly add NaNO2 (12.57 g, 182.20 mmol, 6.2 equivalents) dissolved in H2O (62.5 mL). Allow the mixture to stir at -5 °C for 30 minutes. Add a solution of CuBr (52.70 g, 367.34 mmol, 11.19 mL, 12.5 equivalents) dropwise to an aqueous solution of HBr (260.75 g, 1.06 mol, 175.00 mL, 33% purity, 36.19 equivalents) at 0 °C. Allow the mixture to stir at 70 °C for 30 minutes. The reactants were quenched by adding water (200 mL) and extracted with DCM (200 mL × 3). The organic solutions were combined, washed with aqueous NaOH solution (4 M, 150 mL × 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1). 1-Bromo-3-fluoro-4-methyl-2-nitrobenzene was given as a yellow oil (79.97% yield). 1 H NMR (400 MHz, CDCl3) δ=7.36(dd, J =1.4, 8.3Hz, 1H), 7.26-7.20 (m, 1H), 2.33 (d, J =2.1 Hz, 3H).

[0234] Preparation of 2-bromo-2-(1-(tert-butyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate Step 1: 3-Bromo-N-(tert-butyl)-2-nitroaniline Cs₂CO₃ (148.10 g, 454.55 mmol, 2 equivalents) and t-BuNH₂ (16.62 g, 227.28 mmol, 23.88 mL, 1 equivalent) were added to a solution of 1-bromo-3-fluoro-2-nitrobenzene (50 g, 227.28 mmol, 1 equivalent) in DMSO (100 mL) at 25 °C. The mixture was stirred at 60 °C for 12 hours. TLC (petroleum ether:ethyl acetate = 5:1, R f=0.63) indicates complete depletion of the starting material, and a new point with lower polarity was detected. The mixture was quenched with H2O (300 mL) and ethyl acetate (150 mL) was added to the mixture. The layers were separated and the aqueous phase was extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with brine (300 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give 3-bromo-N-(tert-butyl)-2-nitroaniline as a red oil (64.4% yield). 1 ¹H NMR (400 MHz, methanol-d⁴) δ = 7.21–7.16 (m, 2H), 6.98 (dd, J =2.4, 6.5Hz, 1H), 1.39 (s, 9H).

[0235] Step 2: 3-Bromo-N 1 -(tert-butyl)phenyl-1,2-diamine H₂O (25 mL) was added to a solution of 3-bromo-N-(tert-butyl)-2-nitroaniline (30 g, 109.84 mmol, 1 equivalent) in CH₃COOH (75 mL) and EtOAc (225 mL) at 25 °C. The mixture was heated at 50 °C. Then, Fe (30.67 g, 549.20 mmol, 5 equivalent) was added fractionally at 50 °C, and the mixture was heated to 80 °C and then stirred for 16 hours. TLC (petroleum ether:ethyl acetate = 5:1, R f = 0.61) indicates complete depletion of the starting material, and some new spots with high polarity were detected. The reaction mixture was filtered at 80°C and then concentrated to obtain the residue. The residue was dissolved in EtOAc (200 mL) and the pH was adjusted to 7-8 with saturated NaHCO3. The organic phase was separated and washed with brine (100 mL × 3). The organic phase was dried over Na2SO4, filtered, and concentrated to obtain the crude product. The residue was purified by rapid silica gel chromatography (ISCO®; 200 g SepaFlash® silica gel fast column, eluent 0-50% ethyl acetate / petroleum gradient, @150 mL / min). 3-Bromo-N was obtained as a yellow oil. 1 -(tert-butyl)phenyl-1,2-diamine (74.9% yield). 1 ¹H NMR (400 MHz, methanol-d⁴) δ = 7.00 (dd, J =1.3, 8.0Hz, 1H), 6.88 (dd, J=1.3, 7.9Hz, 1H), 6.52 (t, J =7.9Hz, 1H), 1.27 (s, 9H).

[0236] Step 3: 4-Bromo-1-(tert-butyl)-1,3-dihydro-2H-benzo[d]imidazol-2-one 3-Bromo-N in THF (200 mL) 1 A solution of (tert-butyl)phenyl-1,2-diamine (20 g, 82.26 mmol, 1 equivalent) was supplemented with TEA (24.97 g, 246.77 mmol, 34.35 mL, 3 equivalents) and bis(trichloromethyl) carbonate (24.41 g, 82.26 mmol, 1 equivalent). The mixture was stirred at 20 °C for 16 hours. LCMS (R t = 0.675 min) showed complete depletion of the starting material and a main peak with the desired mass was detected. The reactants were slowly quenched with ice water (200 mL), adjusted to pH=7, and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with brine (200 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by rapid silica gel chromatography (ISCO®; 200 g SepaFlash® silica gel column, eluent 0–50% ethyl acetate / petroleum gradient, 150 mL / min). 4-Bromo-1-(tert-butyl)-1,3-dihydro-2H-benzo[d]imidazol-2-one (84.0% yield) was given as a yellow oil. LCMS: [M+1]=270.9. 1 H NMR (400 MHz, methanol-d4) δ=7.45 (d, J =8.3Hz, 1H), 7.15 (d, J =8.2 Hz, 1H), 6.91 (t, J =8.3Hz, 1H), 1.77 (s, 9H).

[0237] Step 4: 4-Bromo-1-(tert-butyl)-3-methyl-1,3-dihydro-2H-benzo[d]imidazol-2-one NaH (5.53 g, 138.22 mmol, 60% purity, 2 equivalents) was added to a solution of 4-bromo-1-(tert-butyl)-1,3-dihydro-2H-benzo[d]imidazol-2-one (18.6 g, 69.11 mmol, 1 equivalent) in DMF (190 mL). The mixture was stirred at 0 °C for 0.5 h. Then MeI (19.62 g, 138.22 mmol, 62.42 mL, 2 equivalents) was added to the mixture, and the mixture was stirred at 20 °C for 1 h. LCMS (R t = 1.661 min) showed complete depletion of the starting material and a main peak with the desired mass was detected. The reaction mixture was quenched at 0 °C by adding NH4Cl (200 mL) and extracted with EtOAc (150 mL × 3). The combined organic layers were washed with 450 mL brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by rapid silica gel chromatography (ISCO®; 200 g SepaFlash® silica gel column, eluent 0–50% ethyl acetate / petroleum gradient, 150 mL / min). 4-Bromo-1-(tert-butyl)-3-methyl-1,3-dihydro-2H-benzo[d]imidazol-2-one (97.09% yield) was given as a yellow oil. LCMS: [M+1] = 283.1. 1 ¹H NMR (400MHz, methanol-d⁴) δ=7.52 (d, J =8.3Hz, 1H), 7.21 (d, J =8.1 Hz, 1H), 6.92 (t, J =8.3Hz,1H), 3.70 (s, 3H), 1.77 (s, 9H).

[0238] Step 5: 2-(1-(tert-butyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate Pd(t-Bu3P)2 (1.71 g, 6.71 mmol, 0.05 equivalent) was added to a solution of 4-bromo-1-(tert-butyl)-3-methyl-1,3-dihydro-2H-benzo[d]imidazol-2-one (19 g, 67.10 mmol, 1 equivalent) and bromo-(2-tert-butoxy-2-oxo-ethyl)zinc (1 M, 167.75 mL, 2.5 equivalent) in THF (200 mL). The mixture was stirred at 80 °C for 2 hours. TLC (petroleum ether:ethyl acetate = 5:1, R f= 0.43) indicates complete depletion of the starting material and detection of a new spot with high polarity. The reactants were slowly quenched with ice water (200 mL) and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with brine (200 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by rapid silica gel chromatography (ISCO®; 200 g SepaFlash® silica gel column, eluent 0-50% ethyl acetate / petroleum gradient, 120 mL / min). 2-(1-(tert-butyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate (93.61% yield) was obtained as a yellow oil. 1 ¹H NMR (400 MHz, methanol-d⁴) δ=7.47 (dd, J =0.7, 8.2Hz, 1H), 7.02-6.94 (m, 1H), 6.89 (d, J =7.5Hz, 1H), 3.91 (s, 2H), 3.57 (s, 3H), 1.79 (s, 9H), 1.44 (s, 9H).

[0239] Step 6: 2-Bromo-2-(1-(tert-butyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate LDA (2 M, 14.13 mL, 3 equivalents) was added to a solution of 2-(1-(tert-butyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl) tert-butyl acetate (3 g, 9.42 mmol, 1 equivalent) in THF (30 mL) at -70 °C, and the mixture was stirred at -70 °C for 45 min. TMSCl (3.07 g, 28.27 mmol, 3.59 mL, 3 equivalents) was added. After 15 min, a solution of NBS (5.03 g, 28.27 mmol, 3 equivalents) in THF (15 mL) was added, and the mixture was stirred at -70 °C for 1 h. TLC (petroleum ether:ethyl acetate = 5:1, R f= 0.69) indicated complete depletion of the starting material, and a new spot with lower polarity was detected. The reactants were slowly quenched with ice water (30 mL) and extracted with ethyl acetate (15 mL × 2). The combined organic phases were washed with brine (30 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by rapid silica gel chromatography (ISCO®; 20 g SepaFlash® silica gel column, eluent 0–50% ethyl acetate / petroleum gradient, 80 mL / min). 2-Bromo-2-(1-(tert-butyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate (90.83% yield) was obtained as a yellow oil.

[0240] The intermediates in the table below were prepared as follows: from the starting materials shown, purified using appropriate reagents and under appropriate conditions, according to a procedure similar to that described for 2-bromo-2-(1-(tert-butyl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate: .

[0241] Preparation of tert-butyl 2-bromo-2-(3,6-dimethyl-1-(1-methylcyclopropyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)acetate Step 1: 2-(3-methyl-1-(1-methylcyclopropyl)-2-oxo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-benzo[d]imidazol-4-yl)acetate dtbpy (101.80 mg, 379.27 μmol, 0.06 equivalent) and [Ir(OMe)(cod)]2 (251.41 mg, 379.27 μmol, 0.06 equivalent) and Pin2B2 (3.21 g, 12.64 mmol, 2 equivalent) were added to a solution of 2-(3-methyl-1-(1-methylcyclopropyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl) tert-butyl acetate (2 g, 6.32 mmol, 1 equivalent) in dioxane (40 mL) under N2. The mixture was heated to 80 °C and stirred at 80 °C under N2 for 16 hours. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.52) indicates that the starting material was completely exhausted and a major new spot with low polarity was detected. The reaction mixture was filtered and concentrated under reduced pressure to give the residue to give the crude product. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). 2-(3-methyl-1-(1-methylcyclopropyl)-2-oxo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate (4.5 g, crude) was obtained as a yellow oil.

[0242] Step 2: 2-(3,6-dimethyl-1-(1-methylcyclopropyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate K₂CO₃ (3.28 g, 23.74 mmol, 3 equivalents), Pd(PPh₃)₄ (457.15 mg, 395.61 μmol, 0.05 equivalents), and MeI (5.62 g, 39.56 mmol, 2.46 mL, 5 equivalents) were added sequentially to a solution of 2-(3-methyl-1-(1-methylcyclopropyl)-2-oxo-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate (3.5 g, 7.91 mmol, 1 equivalent) in toluene (35 mL), EtOH (14 mL), and H₂O (14 mL) at 20 °C under N₂ conditions. The reaction mixture was then heated to 100°C and stirred at 100°C under N2 for 16 hours. LCMS (R t= 0.797, [M+1]=331.1, 56.55%) indicates complete exhaustion of compound 2, with 56.55% of the desired MS detected. The reaction mixture was quenched with water (150 mL) at 20 °C and extracted with ethyl acetate (30 mL × 3). The combined organic layers were washed with brine (30 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 0 to 80 / 20). 2-(3,6-dimethyl-1-(1-methylcyclopropyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate (42.08% yield) was given as a yellow oil.

[0243] Step 3: 2-Bromo-2-(3,6-dimethyl-1-(1-methylcyclopropyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate LDA (2 M, 2.50 mL, 3 equivalents) was added to a solution of 2-(3,6-dimethyl-1-(1-methylcyclopropyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate (550 mg, 1.66 mmol, 1 equivalent) in THF (10 mL). The mixture was then stirred at -70°C under N2 for 0.5 h. TMSCl (542.52 mg, 4.99 mmol, 633.78 μL, 3 equivalents) was then added to the above reaction mixture at -70°C under N2. The mixture was then stirred at -70°C under N2 for 0.5 h. A solution of NBS (888.79 mg, 4.99 mmol, 3 equivalents) in THF (10 mL) was then added to the above reaction mixture at -70°C under N2. The mixture was then stirred at -70°C under N2 for 1 hour. LCMS showed complete depletion of the starting material and detection of 87.45% of the desired MS. The reaction mixture was quenched with 100 mL of saturated aqueous NH4Cl solution and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). 2-Bromo-2-(3,6-dimethyl-1-(1-methylcyclopropyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate (77.79% yield) was given as a yellow oil. LCMS: [M+1] = 411.1. 1HNMR (400 MHz, CDCl3) δ=7.15 (s, 1H), 6.99 (d, J =0.8 Hz, 1H), 5.94 (s, 1H), 3.71 (s, 3H), 2.41 (s, 3H), 2.05 (s, 2H), 1.49 (s, 9H), 1.15-1.12 (m, 2H), 1.00-0.96 (m, 2H), 0.21-0.13 (m, 1H).

[0244] The intermediates in the table below were prepared as follows: from the starting materials shown, purified using appropriate reagents and under appropriate conditions, according to a procedure similar to that described for 2-bromo-2-(3,6-dimethyl-1-(1-methylcyclopropyl)-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)tert-butyl acetate: .

[0245] Preparation of 4-bromo-6-methylindoline-2-one Step 1: Methyl 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxoacetate LDA (2 M, 317.42 mL, 1.2 equivalents) was added dropwise to a solution of 1-bromo-3-fluoro-5-methylbenzene (100 g, 529.03 mmol, 1 equivalent) in THF (500 mL) at -70 °C under N2. The reaction mixture was then stirred at -70 °C for 0.5 h. A solution of dimethyl oxalate (81.21 g, 687.74 mmol, 1.3 equivalents) in THF (500 mL) was then added to the above reaction mixture at -70 °C under N2. The reaction mixture was then stirred at -70 °C under N2 for 1 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.43) indicates that the starting material was completely exhausted and a new spot was formed. The residue was quenched with aqueous NH4Cl solution (1000 mL) and extracted with EtOAc (500 mL × 3). The combined organic layers were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1). Methyl 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxoacetate was given as a red oil (65.28% yield). 1¹H NMR (400 MHz, chloroform-d) δ=7.29 (s, 1H), 6.96 (d, J=10.3Hz, 1H), 3.95 (s, 3H), 2.40 (s, 3H).

[0246] Step 2: 4-Bromo-1-(tert-butyl)-6-methylindoline-2,3-dione n-BuLi (2.5 M, 30.54 mL, 2.1 equivalents) was added to a solution of t-BuNH2 (5.85 g, 79.98 mmol, 8.40 mL, 2.2 equivalents) in THF (100 mL) at -70 °C under N2. The mixture was stirred at -70 °C for 0.5 h. Methyl 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxoacetate (10 g, 36.35 mmol, 1 equivalent) was added to THF (100 mL) at -70 °C under N2. The mixture was stirred at 20 °C for 2 h. LCMS showed complete depletion of the starting material and a main peak of the desired mass was detected. The reaction mixture was quenched with an aqueous solution of HCl (200 mL) in ice water and extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The crude product was triturated at 20 °C with PE:EA = 3:1 for 30 minutes. 4-Bromo-1-(tert-butyl)-6-methylindoline-2,3-dione was obtained as a red solid (63.47% yield). LCMS: [M+1] = 296.0. 1 ¹H NMR (400 MHz, chloroform-d) δ=7.07 (s, 2H), 2.42 (s, 3H), 1.74 (s, 9H).

[0247] Step 3: 4-Bromo-1-(tert-butyl)-6-methylindoline-2-one NH₂NH₂·H₂O (82.56 g, 1.62 mol, 80 mL, 98% purity, 12.27 equivalents) was added to a solution of 4-bromo-1-(tert-butyl)-6-methylindoline-2,3-dione (39 g, 131.69 mmol, 1 equivalent) in ethylene glycol (400 mL) at 25 °C under N₂. The mixture was stirred at 130 °C for 3 h. LCMS showed complete depletion of the starting material and a main peak of the desired mass was detected. The reaction mixture was concentrated under reduced pressure and quenched by adding H₂O (200 mL), and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was used directly without further purification. 4-Bromo-1-(tert-butyl)-6-methylindololin-2-one was given as a red solid (94.19% yield). LCMS: [M+1] = 284.0. 1 ¹H NMR (400MHz, chloroform-d) δ=6.99 (d, J=9.7Hz, 2H), 3.38 (s, 2H), 2.36 (s, 3H), 1.71 (s, 9H).

[0248] Step 4: 4'-Bromo-1'-(tert-butyl)-6'-methylspiro[cyclopropane-1,3'-indoline]-2'-one NaH (17.86 g, 446.53 mmol, 60% purity, 3 equivalents) was added to a solution of 4-bromo-1-(tert-butyl)-6-methylindoline-2-one (42 g, 148.84 mmol, 1 equivalent) in DMF (400 mL) at 0 °C under N2, and the solution was stirred at 0 °C for 0.5 h. Then, 1,2-dibromoethane (83.89 g, 446.53 mmol, 33.69 mL, 3 equivalents) was added to the solution at 0 °C, and the solution was stirred at 25 °C for 1 h. LCMS showed complete depletion of the starting material and a main peak with the desired mass was detected. The reactants were quenched with saturated NH4Cl (400 mL) and extracted with EtOAc (100 mL × 3). The combined organic layers were washed with brine (100 mL × 3), dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1). 4'-Bromo-1'-(tert-butyl)-6'-methylspiro[cyclopropane-1,3'-indoline]-2'-one was given as a yellow oil (47.96% yield). LCMS: [M+1] = 310.1. 1¹H NMR (400 MHz, chloroform-d) δ=7.03 (s, 1H), 6.85 (s, 1H), 2.28 (s, 3H), 2.15 (q, J=3.7Hz, 2H), 1.67 (s, 9H), 1.35 (q, J=3.7Hz, 2H), 1.31 (s, 3H).

[0249] Step 5: 4-Bromo-6-methylindoline-2-one A solution of 4'-bromo-1'-(tert-butyl)-6'-methylspiro[cyclopropane-1,3'-indoline]-2'-one (22 g, 71.38 mmol, 1 equivalent) in TFA (100 mL) under N2 was added, and the solution was stirred at 25 °C for 0.5 h. LCMS showed complete depletion of the starting material and a main peak with the desired mass was detected. The reactants were concentrated under reduced pressure to obtain the residue. The crude product was milled with MeCN at 20 °C for 30 min. 4-bromo-6-methylindoline-2-one (77.80% yield) was given as a yellow solid. LCMS: [M+1] = 253.9. 1 H NMR (400 MHz, chloroform-d) δ=6.94 (s, 1H), 6.76 (s, 1H), 2.33 (s, 3H), 2.28 (d, J=3.9Hz, 2H), 1.56 (d, J=3.9Hz, 2H).

[0250] Preparation of 5-bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochromane-1,3'-pyran] Step 1: 2-(2,6-dibromophenyl)ethylene oxide 2,6-Dibromobenzaldehyde (30 g, 113.67 mmol, 1 equivalent) and Me3S were reacted in THF (500 mL) and DMSO (100 mL) at -10 °C under N2. + I -NaH (13.64 g, 341.02 mmol, 60% purity, 3 equivalents) was added to a solution of (69.59 g, 341.02 mmol, 3 equivalents). The mixture was stirred at 20 °C for 1 hour. TLC (petroleum ether / ethyl acetate = 10 / 1, R) was performed. f = 0.6) indicated that 2,6-dibromobenzaldehyde was consumed, and a major new spot with high polarity was detected. The mixture was quenched with aqueous NH4Cl solution (1.5 L) and extracted with ethyl acetate (500 mL × 3). The combined organic layers were washed with brine (1000 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The crude product was milled with EtOH at 20 °C for 10 min to give 2-(2,6-dibromophenyl)ethylene oxide as a white solid (66.47% yield). 1 ¹H NMR (400 MHz, chloroform-d) δ=7.55 (d, J=8.1 Hz, 2H), 7.06 (t, J=8.1 Hz, 1H), 3.94-3.85 (m, 1H), 3.33 (dd, J=4.3, 5.0 Hz, 1H), 2.98 (dd, J=2.8, 5.1 Hz, 1H).

[0251] Step 2: 2-(2,6-dibromophenyl)prop-1-ol A solution of 2-(2,6-dibromophenyl)ethylene oxide (46 g, 165.50 mmol, 1 equivalent) was added dropwise to a solution of AlMe3 (2.0 M, 82.75 mL, 1 equivalent) in toluene (500 mL) under N2 conditions. The mixture was stirred at 0 °C for 1 hour. TLC (petroleum ether / ethyl acetate = 3 / 1, Rf = 0.3) indicated that 2-(2,6-dibromophenyl)ethylene oxide was consumed and a major new spot with high polarity was detected. The mixture was quenched by adding Na2SO4·10H2O (100 g) and THF (500 mL). The mixture was filtered and the filtrate was concentrated. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1 to 3 / 1) to give 2-(2,6-dibromophenyl)prop-1-ol as a colorless oil (61.66% yield).

[0252] Step 3: 1,3-Dibromo-2-(1-(methoxymethoxy)propyl-2-yl)benzene MOMBr (23.80 g, 190.49 mmol, 15.55 mL, 2 equivalents) was added to a solution of 2-(2,6-dibromophenyl)prop-1-ol (28 g, 95.24 mmol, 1 equivalent) and DIEA (49.24 g, 380.97 mmol, 66.36 mL, 4 equivalents) in DCM (200 mL), and the mixture was stirred at 20 °C for 16 h. TLC (petroleum ether / ethyl acetate = 1 / 1, Rf = 0.59) indicated that 2-(2,6-dibromophenyl)prop-1-ol was consumed and a major new spot with low polarity was detected. The reaction mixture was quenched by adding an aqueous solution of NaHCO3 (300 mL) and extracted with DCM (100 mL × 3). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give 1,3-dibromo-2-(1-(methoxymethoxy)propyl-2-yl)benzene as a colorless oil (86.97% yield). 1 H NMR (400 MHz, chloroform-) d ) δ=7.57 (d,J=8.0Hz, 1H), 7.51 (d, J=8.0Hz, 1H), 6.90 (t, J=8.0Hz, 1H), 4.67-4.56 (m,2H), 4.17-4.05 (m, 2H), 3.93 (dd, J=6.8, 8.9Hz, 1H), 3.31 (s, 3H), 1.44 (d, J=7.0Hz, 3H).

[0253] Step 4: 3-(3-bromo-2-(1-(methoxymethoxy)prop-2-yl)phenyl)tetrahydro-2H-pyran-3-ol n-BuLi (2.5 M, 6.15 mL, 1 equivalent) was added dropwise to a solution of 1,3-dibromo-2-(1-(methoxymethoxy)propyl-2-yl)benzene (5.2 g, 15.38 mmol, 1 equivalent) and TMEDA (1.97 g, 16.92 mmol, 2.55 mL, 1.1 equivalent) in THF (60 mL). The mixture was stirred at -60 °C for 0.5 h. Then, dihydro-2H-pyran-3(4H)-one (4.62 g, 46.15 mmol, 3 equivalent) was added dropwise to the system at -60 °C, and the mixture was stirred at 0 °C for 0.5 h. TLC (petroleum ether / ethyl acetate = 10 / 1, Rf = 0.4) indicated that 1,3-dibromo-2-(1-(methoxymethoxy)propyl-2-yl)benzene was consumed and a major new spot with high polarity was detected. The mixture was quenched with aqueous NH4Cl solution (300 mL) and extracted with ethyl acetate (200 mL × 2). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 3 / 1 to 2 / 1) to give 3-(3-bromo-2-(1-(methoxymethoxy)propyl-2-yl)phenyl)tetrahydro-2H-pyran-3-ol (37.09% yield) as a colorless oil.

[0254] Step 5: 3-(3-bromo-2-(1-hydroxypropyl-2-yl)phenyl)tetrahydro-2H-pyran-3-ol HCl (100 mL) was added to a solution of 3-(3-bromo-2-(1-(methoxymethoxy)prop-2-yl)phenyl)tetrahydro-2H-pyran-3-ol (14 g, 38.97 mmol, 1 equivalent) in MeCN (50 mL) at 20 °C, and the mixture was stirred at 20 °C for 12 h. TLC (petroleum ether / ethyl acetate = 3 / 1, Rf = 0.42) indicated that 3-(3-bromo-2-(1-(methoxymethoxy)prop-2-yl)phenyl)tetrahydro-2H-pyran-3-ol was consumed, and a major new spot with high polarity was detected. The mixture was quenched with H₂O (10 mL) and extracted with ethyl acetate (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 3 / 1 to 1 / 1) to give 3-(3-bromo-2-(1-hydroxypropyl-2-yl)phenyl)tetrahydro-2H-pyran-3-ol as a yellow oil (10.7 g, 23.76 mmol, 60.98% yield, 70% purity). LCMS: [M+1] = 296.9.

[0255] Step 6: 5-Bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran] CMBP (8.60 g, 35.64 mmol, 1.5 equivalents) was added to a solution of 3-(3-bromo-2-(1-hydroxypropyl-2-yl)phenyl)tetrahydro-2H-pyran-3-ol (10.7 g, 23.76 mmol, 1 equivalent) in toluene (100 mL) at 20 °C. The mixture was stirred at 80 °C for 16 h. LCMS showed that 3-(3-bromo-2-(1-hydroxypropyl-2-yl)phenyl)tetrahydro-2H-pyran-3-ol was consumed and approximately 53% of the desired compound was detected. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1) and by preparative HPLC (column: Phenomenex luna c18 250 mm). 100mm 10 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 30%-65% B (purified for 24.0 min) to give 5-bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran] as a yellow oil (67.97% yield).

[0256] Step 7: Stereoisomers of 5-bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran] 5-Bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran] (4.8 g, 16.15 mmol, 1 equivalent) was obtained by SFC (column: DAICL CHIRALPAK AD (250 mm)). Separation was performed using a mobile phase of [CO2-MeOH] (30 mm, 10 μm) and a B% elution mode (15% B, isocratic elution mode) to obtain four isomers.

[0257] Peak 1 was obtained as a white solid, arbitrarily designated as (1S, 4S)-5-bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran] (20.83% yield). LCMS: [M+1] = 297.2. SFC: Rt = 1.656. 1 H NMR (400 MHz, chloroform-d) δ=7.46 (d, J=7.5Hz, 1H), 7.13-7.06 (m, 2H), 4.08 (brdd, J=4.2, 11.2 Hz, 1H), 4.01 (brd, J=12.4Hz, 1H), 3.96-3.91 (m, 1H), 3.87-3.82 (m,1H), 3.56-3.49 (m, 1H), 3.39 (d, J=12.5Hz, 1H), 3.00 (q, J=6.2 Hz, 1H), 2.30-2.19 (m, 1H), 2.06-1.97 (m, 2H), 1.57 (brd, J=9.3Hz, 1H), 1.39 (d, J=6.9Hz, 3H).

[0258] Peak 2 was obtained as a white solid and arbitrarily designated as (1S,4R)-5-bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran] (16.88% yield). LCMS: [M+1] = 297.2. SFC: Rt = 1.848. 1H NMR (400 MHz, chloroform-d) δ=7.46 (dd, J=2.3, 6.6Hz, 1H), 7.11-7.06 (m, 2H), 4.08 (brdd, J=3.0, 12.4Hz, 1H), 3.91-3.85 (m, 2H), 3.76-3.74 (m, 1H), 3.71-3.67(m, 1H), 3.54 (brt, J=11.8 Hz, 1H), 3.05-2.98 (m, 1H), 2.24-2.14 (m, 2H),1.79-1.71 (m, 1H), 1.51 (brd, J=13.5Hz, 1H), 1.39 (d, J=6.9Hz, 3H).

[0259] Peak 3 was obtained as a white solid and arbitrarily designated as (1R,4R)-5-bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran] (22.92% yield). LCMS: [M+1] = 297.2. SFC: Rt = 2.441. 1 H NMR(400 MHz, chloroform-d) δ=7.46 (dd, J=1.6, 7.4Hz, 1H), 7.12-7.05 (m, 2H), 4.08(brdd, J=4.4, 11.2 Hz, 1H), 4.02 (brd, J=12.4Hz, 1H), 3.96-3.92 (m, 1H), 3.87-3.83 (m, 1H), 3.56-3.49 (m, 1H), 3.39 (d, J=12.4Hz, 1H), 3.03-2.97 (m, 1H), 2.29-2.19 (m, 1H), 2.06-1.98 (m, 2H), 1.56 (brs, 1H), 1.39 (d, J=6.9Hz, 3H).

[0260] Peak 4 was obtained as a white solid and arbitrarily designated as (1R,4S)-5-bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran] (22.92% yield). LCMS: [M+1] = 297.2. SFC: Rt = 2.441. 1H NMR(400 MHz, chloroform-d) δ=7.46 (dd, J=2.2, 6.7Hz, 1H), 7.11-7.06 (m, 2H), 4.11-4.06(m, 1H), 3.90-3.85 (m, 2H), 3.74 (brd, J=2.0Hz, 1H), 3.71-3.67 (m, 1H), 3.57-3.51 (m, 1H), 3.01 (q, J=6.8 Hz, 1H), 2.24-2.14 (m, 2H), 1.79-1.71 (m, 1H),1.56-1.49 (m, 1H), 1.39 (d, J=6.9Hz, 3H).

[0261] Preparation of 5-bromo-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran] The isomers of 5-bromo-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran] were prepared using a method similar to that described above for the preparation of 5-bromo-4-methyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran], and were obtained by SFC (column: DAICEL CHIRALPAKAD (250 mm)). Separation was performed using a mobile phase of [CO2-EtOH] (30 mm, 10 μm) and a B% elution mode of 15%.

[0262] Peak 1, obtained as a white solid, was arbitrarily designated as (S)-5-bromo-4-methyl-2',3',5',6'-tetrahydrospiro-[isochroman-1,4'-pyran]. SFC R t =1.638 min. 1 H NMR (400 MHz, CDCl3) δ=7.44 (dd, J =1.6,7.4Hz, 1H), 7.14-7.07 (m, 2H), 3.99-3.92 (m, 1H), 3.90-3.86 (m, 2H), 3.85-3.82 (m, 2H), 3.80 (s, 1H), 3.04-2.96 (m, 1H), 2.23-2.16 (m, 1H), 1.93-1.87(m, 2H), 1.65 (brd, J =13.5Hz, 1H), 1.38 (d, J =6.9Hz, 3H).

[0263] Peak 2, obtained as a white solid, was arbitrarily designated as (R)-5-bromo-4-methyl-2',3',5',6'-tetrahydrospiro-[isochroman-1,4'-pyran]. SFC R t =1.873 min. 1 H NMR (400 MHz, CDCl3) δ=7.44 (dd, J =1.6,7.4Hz, 1H), 7.15-7.08 (m, 2H), 3.99-3.92 (m, 1H), 3.87 (dd, J =4.7, 7.1 Hz,2H), 3.85-3.83 (m, 2H), 3.80 (s, 1H), 3.02-2.96 (m, 1H), 2.25-2.17 (m, 1H),1.92-1.87 (m, 2H), 1.65 (brd, J =13.6Hz, 1H), 1.38 (d, J =6.9Hz, 3H).

[0264] Preparation of methyl 2-bromo-2-((S)-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetate Step 1: (S)-5-Allyl-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran] At 20 °C, 2-allyl-4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentane (791.61 mg, 4.71 mmol, 2 equivalents), Pd(dppf)Cl2 (172.35 mg, 235.54 μmol, 0.1 equivalents), and Cs2CO3 (1.53 g, 4.71 mmol, 2 equivalents) were added to a solution of (S)-5-bromo-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran] (0.7 g, 2.36 mmol, 1 equivalent) in dioxane (10 mL). The mixture was allowed to be stirred at 80 °C under a N2 atmosphere for 16 hours. The reaction mixture was quenched by the addition of H2O (50 mL) and extracted with ethyl acetate (10 mL × 3). The organic solutions were combined, washed with brine (20 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give (S)-5-allyl-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran] as a white solid (0.62 g, 1.20 mmol, 50.94% yield, 50% purity).

[0265] Step 2: (S)-2-(4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetic acid RuCl3 (24.09 mg, 116.12 μmol, 7.74 μL, 0.1 equivalent) and NaIO4 (993.47 mg, 4.64 mmol, 257.38 μL, 4 equivalent) were added to a solution of (S)-5-allyl-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran] (0.6 g, 1.16 mmol, 1 equivalent) in MeCN (5 mL), hexane (5 mL), and H2O (10 mL) at -10 °C. The mixture was allowed to be stirred at -10 °C for 1 hour. The reaction mixture was quenched by slow addition of Na2SO3 aqueous solution (100 mL), adjusted to pH 5, and extracted with ethyl acetate (50 mL × 2). The organic solutions were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum to give (S)-2-(4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetic acid as a white solid (0.5 g, 904.73 μmol, 77.91% yield, 50% purity).

[0266] Step 3: (S)-2-(4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)methyl acetate H₂SO₄ (460.00 mg, 4.69 mmol, 250.00 μL, 5.18 equivalents) was added to a solution of (S)-2-(4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetic acid (0.5 g, 904.73 μmol, 1 equivalent) in MeOH (10 mL) at 20 °C. The mixture was allowed to be stirred at 70 °C for 16 h. The reaction mixture was quenched by adding NaHCO₃ (50 mL) and extracted with ethyl acetate (20 mL × 3). The organic solutions were combined, washed with brine (20 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give (S)-2-(4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetate as a colorless oil (95.17% yield). 1 H NMR (400 MHz, CDCl3) δ=7.23-7.18 (m, 1H), 7.16-7.09(m, 2H), 3.99-3.91 (m, 1H), 3.90-3.81 (m, 4H), 3.78 (d, J =8.2 Hz, 1H), 3.76-3.72 (m, 1H), 3.71 (s, 3H), 3.65-3.59 (m, 1H), 2.87 (q, J =6.6Hz, 1H), 2.22(dt, J =5.3, 13.1 Hz, 1H), 1.95-1.89 (m, 2H), 1.66 (brd, J =13.6Hz, 1H), 1.33 (d, J =6.9Hz, 3H).

[0267] Step 4: Methyl 2-bromo-2-((S)-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetate LDA (2 M, 861.02 μmol, 2 equivalents) was added to a solution of (S)-2-(4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetate (0.25 g, 861.02 μmol, 1 equivalent) in THF (3 mL) at -60 °C, and the mixture was stirred at -60 °C for 30 min. TMSCl (205.79 mg, 1.89 mmol, 240.41 μL, 2.2 equivalents) was added at -60 °C, and the mixture was stirred at -60 °C for 0.5 h. A solution of NBS (199.22 mg, 1.12 mmol, 1.3 equivalents) in THF (3 mL) was added at -60 °C. The mixture was stirred at -60 °C for 1 h. The mixture was diluted with NH4Cl (50 mL) and extracted with ethyl acetate (20 mL × 3). The organic solutions were combined, washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give methyl 2-bromo-2-((S)-4-methyl-2',3',5',6'-tetrahydrospiro-[isochroman-1,4'-pyran]-5-yl)acetate as a yellow solid (78.63% yield). 1 H NMR (400 MHz, CDCl3) δ=7.59(d, J =7.8 Hz, 1H), 7.31-7.28 (m, 1H), 7.18 (d, J =8.0Hz, 1H), 5.69 (s, 1H), 3.94(brd, J =12.3Hz, 1H), 3.86 (brdd, J =3.0, 10.6Hz, 4H), 3.83 (s, 3H), 3.80-3.78(m, 1H), 2.93-2.87 (m, 1H), 2.19 (dt, J =5.4, 12.9Hz, 1H), 1.95-1.91 (m, 2H),1.65 (brd, J =14.0Hz, 1H), 1.45 (d, J =7.0Hz, 3H).

[0268] Methyl 2-bromo-2-((R)-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetate was prepared from (R)-5-bromo-4-methyl-2',3',5',6'-tetrahydrospiro[isochroman-1,4'-pyran]-5-yl)acetate.

[0269] Preparation of methyl 2-bromo-2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate Step 1: 4'-Bromospiro[cyclopropane-1,3'-indoline]-2'-one (2-bromoethyl)diphenylsulfonium trifluoromethanesulfonate (18.82 g, 42.44 mmol, 1.5 equivalent) was added to a solution of 4'-bromospiro[cyclopropane-1,3'-indoline]-2'-one (6 g, 28.30 mmol, 1 equivalent) in DMF (250 mL) under N2 at 25 °C, and the solution was stirred at 25 °C for 6 min. Then, TEA (8.59 g, 84.89 mmol, 11.82 mL, 3 equivalent) was added to the solution at 25 °C, and the solution was stirred at 25 °C for 6 h. LCMS showed complete depletion of the starting material and a peak with the desired mass was detected. The reaction mixture was quenched by adding H2O (750 mL) at 0 °C and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (200 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 3 / 1). 4'-bromospiro[cyclopropane-1,3'-indoline]-2'-one was given as a yellow oil (62.34% yield). 1 H NMR (400 MHz, methanol-d4) δ=7.12-7.06 (m, 2H), 6.96 (dd, J=2.3, 6.4Hz, 1H), 2.29 (q, J=3.9Hz, 2H), 1.47 (q, J=3.9Hz, 2H).

[0270] Step 2: 4'-Bromo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one CMBP (6.89 g, 28.56 mmol, 2 equivalents) and tetrahydro-2H-pyran-4-ol (5.83 g, 57.12 mmol, 5.72 mL, 4 equivalents) were added to a solution of 4'-bromospiro[cyclopropane-1,3'-indoline]-2'-one (3.4 g, 14.28 mmol, 1 equivalent) in toluene (10 mL) at 25 °C under N2, and the solution was stirred at 120 °C for 16 h. LCMS showed that the starting material remained and two peaks with the desired mass were detected. The reaction mixture was quenched by adding H2O (50 mL) at 0 °C and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was analyzed by preparative HPLC (column: Welch Xtimate C18 250). 70 mm # 10 μm; Mobile phase: [water (TFA)-ACN]; B%: 25%-55%, 20 min. HPLC:R t (3.332 min) Further purification was performed to obtain the product. 4'-bromo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one was obtained as a yellow solid (34.77% yield).

[0271] Step 3: Methyl 2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate Pd(t-Bu3P)2 (126.89 mg, 248.30 μmol, 0.05 equivalent) and BrZnCH2COOMe (1 M, 14.90 mL, 3 equivalent) were added to a solution of 4'-bromo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one (1.6 g, 4.97 mmol, 1 equivalent) in THF (15 mL) at 25 °C under N2, and the solution was stirred at 80 °C for 2 hours. LCMS (R t=0.602 min) showed that the starting material remained and two peaks with the desired mass were detected. The reaction mixture was quenched by adding H2O (50 mL) at 0 °C and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (10 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 3 / 1). Methyl 2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (1.6 g, crude) was obtained as a yellow solid.

[0272] Step 4: Methyl 2-bromo-2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate LDA (2 M, 1.71 mL, 1.8 equivalents) was added dropwise to a solution of methyl 2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (600 mg, 1.90 mmol, 1 equivalent) in THF (4 mL) under N2 at -70 °C, and the solution was stirred at -78 °C for 0.5 h. Then, TMSCl (413.40 mg, 3.81 mmol, 482.94 μL, 2 equivalents) was added to the solution at -78 °C, and the solution was stirred at -78 °C for 0.5 h. Then, NBS (677.24 mg, 3.81 mmol, 2 equivalents) in THF (2 mL) was added to the solution at -78 °C, and the solution was stirred at -78 °C for 1 h. LCMS showed the starting material remaining and detected a peak with the desired mass. The reaction mixture was quenched at 0 °C with the addition of NH4Cl (10 mL) and extracted with ethyl acetate (5 mL × 3). The combined organic layers were washed with brine (5 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 3 / 1). Methyl 2-bromo-2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate was given as a yellow oil (79.99% yield).

[0273] The intermediates in the table below were prepared as follows: from the starting materials shown, purified using appropriate reagents and under appropriate conditions, according to a procedure similar to that described for methyl 2-bromo-2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate: .

[0274] Preparation of methyl 2-(6'-methoxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate Step 1: methyl 2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)-6'-(4,4,5,5-tetramethyl-1,3,2-dioxaborphane-2-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate Pin2B2 (483.14 mg, 1.90 mmol, 2 equivalents), [Ir(OMe)(cod)]2 (51.06 mg, 190.26 μmol, 0.2 equivalents), and dtbpy (50.00 mg, 75.43 μmol, 7.93e-2 equivalents) were added to a solution of methyl 2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (300 mg, 951.29 μmol, 1 equivalent) in dioxane (6 mL) under N2. The mixture was heated to 60 °C and stirred at 60 °C under N2 for 2 h. LCMS showed complete depletion of the starting material and 77.62% of the desired Ms was detected. H₂O (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 3 / 1). Methyl 2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)-6'-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (2.3 g, crude) was given as a yellow solid. LCMS: [M+1] = 442.3. 1 ¹H NMR (400 MHz, methanol-d⁴) δ = 7.47 (s, ¹H), 7.34 (s, ¹H), 4.47 (ddd,J =4.2, 8.0, 12.3Hz, 1H), 4.09 (brdd, J =4.2,11.5Hz, 2H), 3.68 (s, 3H), 3.59 (brt, J =11.4Hz, 2H), 3.47 (s, 2H), 2.66-2.55(m, 2H), 2.06-1.99 (m, 2H), 1.68 (brdd, J =2.0, 12.3Hz, 2H), 1.60-1.54 (m, 2H), 1.37 (s, 12H).

[0275] Step 2: Methyl 2-(6'-hydroxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate At 0°C, potassium persulfate preparation (Oxone) (3.36 g, 5.47 mmol, 1.5 equivalent) in H₂O (50 mL) was added to a solution of methyl 2-(2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)-6'-(4,4,5,5-tetramethyl-1,3,2-dioxaborpine-2-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (2.3 g, 3.65 mmol, 1 equivalent) in acetone (100 mL). The mixture was heated to 25°C and stirred at 25°C for 1 hour. LCMS showed complete depletion of the starting material and 86.72% of the desired Ms were detected. The mixture was quenched with Na₂SO₃ (100 mL) and ethyl acetate (100 mL) was added to the mixture. The layers were separated and the aqueous phase was extracted with ethyl acetate (100 mL × 3). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). Methyl 2-(6'-hydroxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate was given as a yellow solid (64.52% yield). LCMS: [M+1] = 332.1. 1 ¹H NMR (400 MHz, chloroform-d) δ=6.74 (d, J =2.1 Hz, 1H), 6.42 (d, J =2.0Hz, 1H), 4.56 (tt, J=4.1, 12.4Hz, 1H), 4.14 (brdd, J =4.4, 11.5Hz, 2H), 3.71 (s, 3H), 3.58-3.51 (m, 2H), 3.31 (s, 2H), 2.60-2.49 (m, 2H), 1.87-1.80 (m, 2H), 1.69(brdd, J =2.3, 12.8 Hz, 2H), 1.64-1.59 (m, 3H).

[0276] Step 3: Methyl 2-bromo-2-(6'-methoxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate MeI (1.75 g, 12.31 mmol, 766.52 μL, 6 equivalents) was added fractionally to a mixture of methyl 2-(6'-hydroxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (680 mg, 2.05 mmol, 1 equivalent) and K2CO3 (425.42 mg, 3.08 mmol, 1.5 equivalents) in DMF (10 mL). The mixture was stirred at 25 °C for 2 hours. LCMS showed approximately 8% of the starting material remaining and detected 87.09% of the desired mass. H2O (20 mL) was added to the reaction mixture and it was extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (20 mL × 1), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 1 / 1). Methyl 2-bromo-2-(6'-methoxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate was given as a yellow solid (73.37% yield). LCMS: [M+1] = 346.1. 1 ¹H NMR (400 MHz, chloroform-d) δ=6.71 (d, J =2.1 Hz, 1H), 6.46 (d, J =2.1 Hz, 1H), 4.52 (tt, J =4.3, 12.4Hz, 1H), 4.12 (dd, J=4.6,11.5Hz, 2H), 3.83 (s, 3H), 3.71 (s, 3H), 3.53 (dt, J =1.9, 12.0Hz, 2H), 3.35(s, 2H), 2.60-2.49 (m, 2H), 1.86-1.81 (m, 2H), 1.68 (brdd, J =2.6, 12.5Hz, 2H),1.63-1.59 (m, 2H).

[0277] Step 4: Methyl 2-(6'-methoxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate A mixture of methyl 2-bromo-2-(6'-methoxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (570 mg, 1.65 mmol, 1 equivalent) in THF (20 mL) was stirred at 25 °C under N2. The mixture was cooled to -70 °C under N2. Then, LDA (2 M, 2.48 mL, 3 equivalents) was added at -70 °C, and the mixture was stirred at -70 °C under N2 for 30 min. Then, TMSCl (537.88 mg, 4.95 mmol, 628.36 μL, 3 equivalents) was added, and the mixture was stirred at -70 °C under N2 for 30 min. A solution of NBS (881.19 mg, 4.95 mmol, 3 equivalents) in THF (10 mL) was then added to the mixture at -70 °C, and the mixture was stirred at -70 °C under N2 for 1 h. LCMS showed 33.57% of the starting material remaining, and 62.09% of the desired mass was detected. The mixture was quenched with NH4Cl (10 mL), and ethyl acetate (10 × 3 mL) was added to the mixture. The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 3 / 1). Methyl 2-(6'-methoxy-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (58.55% yield) was given as a yellow oil. LCMS :[M+1]=426.4. 1¹H NMR (400 MHz, chloroform-d) δ=6.88 (d, J=2.1 Hz, 1H), 6.75 (d, J=2.1 Hz, 1H), 4.96 (s, 1H), 4.56-4.47 (m, 1H), 4.13 (brdd, J=2.0, 9.4Hz, 2H), 3.85 (s, 3H), 3.80 (s, 3H), 3.56-3.49 (m, 2H), 2.56-2.47 (m, 2H), 1.96-1.90 (m, 1H), 1.89-1.84 (m, 1H), 1.75-1.65 (m, 4H).

[0278] Using a procedure similar to that described in steps 1, 2 and 3 above, 7-bromo-5-methoxy-1,3-dimethyl-1H-indazole was prepared as a white solid, starting with 7-bromo-1,3-dimethyl-1H-indazole. 1 H NMR (400 MHz, CD3OD)δ=7.24 (d, J =2.1 Hz, 1H), 7.04 (d, J =2.1 Hz, 1H), 4.23 (s, 3H), 3.83 (s, 3H), 2.46 (s, 3H).

[0279] Using a procedure similar to that described in steps 1, 2 and 3 above, 7-bromo-5-(2-methoxyethoxy)-1,3-dimethyl-1H-indazole was prepared as a yellow solid, starting with 7-bromo-1,3-dimethyl-1H-indazole. 1 H NMR (400MHz, CDCl3) δ=7.31 (d, J =2.1 Hz, 1H), 6.93 (d, J =2.1 Hz, 1H), 4.29 (s, 3H), 4.18-4.12 (m, 2H), 3.82-3.74 (m, 2H), 3.48 (s, 3H), 2.49 (s, 3H).

[0280] Preparation of methyl 2-bromo-2-((S)-6-fluoro-1-methylisochroman-8-yl)acetate and methyl 2-bromo-2-((R)-6-fluoro-1-methylisochroman-8-yl)acetate Step 1: 7-Bromo-5-fluoro-1-methyl-2,3-dihydro-1H-inden-1-ol 7-Bromo-5-fluoro-2,3-dihydro-1H-inden-1-one (20 g, 87.32 mmol, 1 equivalent) was added to a solution of MeMgBr (3 M, 145.53 mL, 5 equivalents) in THF (200 mL) under a N2 atmosphere at 0 °C. The mixture was stirred at 0 °C for 2 hours. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.47) indicates that the starting material has been completely exhausted and a new point with lower polarity has been formed. The reaction mixture was quenched with NH4Cl (500 mL) and extracted with ethyl acetate (200 mL × 3). The combined organic layers were washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give 7-bromo-5-fluoro-1-methyl-2,3-dihydro-1H-inden-1-ol (22.2 g, crude) as a white solid. The crude product was used directly in the next step without purification. 1 ¹H NMR (400 MHz, chloroform-d) δ=7.11 (dd, J =1.9, 8.4Hz, 1H), 6.87 (dd, J =0.9, 8.1 Hz, 1H), 2.97-2.86 (m, 1H), 2.84-2.76 (m, 1H), 2.29 (dd, J =5.6,8.6Hz, 2H), 1.63 (s, 3H).

[0281] Step 2: 4-Bromo-6-fluoro-3-methyl-1H-indene HCl (12 M, 200 mL, 26.50 equivalents) was added to a solution of 7-bromo-5-fluoro-1-methyl-2,3-dihydro-1H-inden-1-ol (22.2 g, 90.58 mmol, 1 equivalent) in THF (200 mL) at 0 °C under a N2 atmosphere. The mixture was stirred at 20 °C for 2 hours. TLC (petroleum ether / ethyl acetate = 5 / 1, R f= 0.69) indicates that the starting material remains and a new point with lower polarity is formed. The reaction mixture was quenched with H2O (400 mL) and extracted with MTBE (200 mL × 4). The combined organic layers were washed with brine (400 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid silica gel chromatography (ISCO®; 220 g SepaFlash® silica gel column, eluent 0–5% ethyl acetate / petroleum ether gradient, @ 90 mL / min). 4-Bromo-6-fluoro-3-methyl-1H-indene (97.24% yield) was given as a white solid. 1 H NMR (400 MHz, chloroform-) d ) δ=7.19 (dd, J =2.3, 8.8 Hz, 1H), 7.10 (dd, J =2.3,8.0Hz, 1H), 6.26-6.22 (m, 1H), 3.29 (t, J =2.0Hz, 2H), 2.40 (q, J =2.0Hz, 3H).

[0282] Step 3: 1-(2-bromo-4-fluoro-6-(2-hydroxyethyl)phenyl)ethanol-1-ol A solution of 4-bromo-6-fluoro-3-methyl-1H-indene (20 g, 88.08 mmol, 1 equivalent) in DCM (200 mL) and MeOH (40 mL) was bubbled with O3 (21.14 g, 440.39 mmol, 5 equivalents) for 2 hours at -70 °C. Then, NaBH4 (11.30 g, 298.58 mmol, 3.39 equivalents) was added at 0 °C, and the mixture was stirred at 25 °C for 16 hours. TLC (petroleum ether / ethyl acetate = 3 / 1, R f = 0.41) indicates that the starting material has been completely exhausted and a new point with higher polarity has been formed. The reaction mixture was quenched with H2O (300 mL) and extracted with DCM (100 mL × 3). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give 1-(2-bromo-4-fluoro-6-(2-hydroxyethyl)phenyl)ethyl-1-ol (20 g, crude) as a white solid. The crude product was used directly in the next step without purification. 1 H NMR (400 MHz, chloroform-) d ) δ=7.21 (dd, J=2.6, 7.7Hz, 1H), 6.95 (dd, J =2.6, 9.2 Hz, 1H), 5.59 (q, J =6.7Hz, 1H), 3.99 (td, J =5.1, 10.2 Hz, 1H), 3.84(dt, J =5.1, 9.7Hz, 1H), 3.64-3.52 (m, 1H), 2.84 (td, J =4.7, 13.7Hz, 1H), 1.55(d, J =6.8 Hz, 3H).

[0283] Step 4: 8-Bromo-6-fluoro-1-methylisochromanine PPh3 (29.91 g, 114.02 mmol, 1.5 equivalent) and DIAD (26.13 g, 129.23 mmol, 25.05 mL, 1.7 equivalent) were added to a solution of 1-(2-bromo-4-fluoro-6-(2-hydroxyethyl)phenyl)ethanol-1-ol (20 g, 76.02 mmol, 1 equivalent) in THF (300 mL) at 0 °C, and the mixture was stirred at 25 °C under a N2 atmosphere for 16 hours. TLC (petroleum ether / ethyl acetate = 10 / 1, R f = 0.57) indicates that the starting material has been completely depleted and two new points with lower polarity have formed. A top point is required. The reaction mixture was quenched with H2O (500 mL) and extracted with ethyl acetate (200 mL × 3). The combined organic layers were washed with brine (500 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid silica gel chromatography (ISCO®; 220 g SepaFlash® silica gel column, eluent gradient of 0–22% ethyl acetate / petroleum ether, @ 80 mL / min). 8-Bromo-6-fluoro-1-methylisochromanol was given as a purple oil (40.79% yield). 1 ¹H NMR (400 MHz, chloroform-d) δ=7.15 (dd, J =2.5, 8.1 Hz, 1H), 6.83 (dd, J =2.5, 8.7Hz, 1H), 5.02 (q, J =6.4Hz, 1H), 4.08 (ddd, J =4.3, 9.6, 11.7Hz, 1H), 3.85 (ddd, J=3.5,6.0, 11.7Hz, 1H), 2.97-2.85 (m, 1H), 2.78-2.67 (m, 1H), 1.57 (d, J =6.4Hz, 3H).

[0284] Step 5: Methyl 2-(6-fluoro-1-methylisochroman-8-yl)acetate Pd(t-Bu3P)2 (1.56 g, 3.06 mmol, 0.1 equivalent) and MeOOCCH2ZnBr (1 M, 45.90 mL, 1.5 equivalent) were added to a solution of 8-bromo-6-fluoro-1-methylisochloromethane (7.5 g, 30.60 mmol, 1 equivalent) in THF (80 mL) at 25 °C. The mixture was stirred at 80 °C under a N2 atmosphere for 2 hours. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.36) indicates that the starting material was completely depleted and three new points with greater polarity were formed. The reaction mixture was quenched with H2O (150 mL). The mixture was filtered and the filtrate was extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid silica gel chromatography (ISCO®; 120 g SepaFlash® silica gel column, eluent gradient of 0–28% ethyl acetate / petroleum ether, @ 80 mL / min). Methyl 2-(6-fluoro-1-methylisochroman-8-yl)acetate was given as a yellow oil (96.01% yield). 1 H NMR (400MHz, chloroform-) d ) δ=6.85 (dd, J =2.6, 9.4Hz, 1H), 6.77 (dd, J =2.3, 8.8 Hz, 1H), 5.06(q, J =6.5Hz, 1H), 4.13-4.05 (m, 1H), 3.82 (td, J =5.4, 10.8 Hz, 1H), 3.72 (s,3H), 3.64-3.46 (m, 2H), 2.99-2.88 (m, 1H), 2.81-2.72 (m, 1H), 1.46 (d, J =6.5Hz, 3H).

[0285] Step 6: (S)-2-(6-fluoro-1-methylisochroman-8-yl)acetate and (R)-2-(6-fluoro-1-methylisochroman-8-yl)acetate 2-(6-fluoro-1-methylisochroman-8-yl)methyl acetate (7 g) was separated by SFC separation (column: ChiRAlPak IH, 25050 mm, 10 μm; mobile phase: [CO2-IPA (0.1%NH3H2O)]; B%: 11%, isocratic elution mode).

[0286] Peak 1 (44.29% yield) was obtained as a yellow oil and arbitrarily designated as methyl(S)-2-(6-fluoro-1-methylisochroman-8-yl). 1 ¹H NMR (400 MHz, chloroform-d) δ=6.85 (dd, J =2.5, 9.5Hz, 1H), 6.77 (dd, J =2.3, 8.9Hz, 1H), 5.06 (q, J =6.5Hz, 1H), 4.08 (ddd, J =4.4, 8.7, 11.5Hz, 1H),3.88-3.78 (m, 1H), 3.72 (s, 3H), 3.65-3.47 (m, 2H), 2.97-2.85 (m, 1H), 2.81-2.72 (m, 1H), 1.46 (d, J =6.5Hz, 3H).

[0287] Peak 2 (45.71% yield) was obtained as a yellow oil and arbitrarily designated as (R)-2-(6-fluoro-1-methylisochroman-8-yl)acetic acid methyl ester. 1 ¹H NMR (400 MHz, chloroform-d) δ=6.85 (dd, J =2.5, 9.4Hz, 1H), 6.77(dd, J =2.3, 8.9Hz, 1H), 5.06 (q, J =6.5Hz, 1H), 4.08 (ddd, J =4.5, 8.8, 11.5Hz,1H), 3.87-3.78 (m, 1H), 3.72 (s, 3H), 3.65-3.45 (m, 2H), 2.99-2.87 (m, 1H),2.82-2.70 (m, 1H), 1.46 (d, J=6.5Hz, 3H).

[0288] Step 7: Methyl 2-bromo-2-((S)-6-fluoro-1-methylisochroman-8-yl)acetate LDA (2 M, 6.30 mL, 3 equivalents) was added to an arbitrary specified solution of methyl(S)-2-(6-fluoro-1-methylisochroman-8-yl) (1 g, 4.20 mmol, 1 equivalent) in THF (20 mL) at -78 °C under a N2 atmosphere. The mixture was stirred at -78 °C for 30 min. TMSCl (1.37 g, 12.59 mmol, 1.60 mL, 3 equivalents) was added at -78 °C. After stirring the reaction mixture at -78 °C for 30 min, NBS (2.24 g, 12.59 mmol, 3 equivalents) was added in THF (40 mL). The mixture was stirred at -78 °C for 1 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.45, 0.51) indicates that the starting material was completely depleted and two new points with lower polarity were formed. The reaction mixture was quenched by adding an aqueous solution of NH4Cl (40 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid silica gel chromatography (ISCO®; 40 g SepaFlash® silica gel column, eluent of 0–22% ethyl acetate / petroleum ether gradient, @ 80 mL / min). Methyl 2-bromo-2-((S)-6-fluoro-1-methylisochroman-8-yl)acetate (97.66% yield) was given as a yellow oil. 1 H NMR (400MHz, chloroform-) d ) δ=7.36 (ddd, J =2.6, 9.7, 16.4Hz, 1H), 6.83 (brd, J =8.6Hz, 1H),5.45-5.35 (m, 1H), 5.23-5.10 (m, 1H), 4.09-4.04 (m, 1H), 3.87-3.77 (m, 4H),3.01-2.88 (m, 1H), 2.81-2.71 (m, 1H), 1.56 (dd, J =6.6, 12.1 Hz, 3H).

[0289] 2-bromo-2-((R)-6-fluoro-1-methylisochroman-8-yl)acetic acid methyl ester is prepared from any specified (R)-2-(6-fluoro-1-methylisochroman-8-yl)acetic acid methyl ester by a procedure similar to that described in step 7 above. 1 H NMR (400 MHz, chloroform-) d ) δ=7.36 (ddd, J =2.6, 9.7, 16.3Hz, 1H), 6.83 (brd, J =8.6Hz, 1H), 5.46-5.35 (m, 1H), 5.22-5.06 (m, 1H), 4.09-4.03 (m, 1H), 3.86-3.77 (m, 4H), 2.99-2.88 (m, 1H), 2.79-2.72 (m, 1H), 1.56 (dd, J =6.6, 12.0Hz, 3H).

[0290] 2-bromo-2-((R)-1-methylisochroman-8-yl)acetic acid methyl ester is prepared by a procedure similar to that described in step 7 above, starting with any specified (R)-2-(1-methylisochroman-8-yl)acetic acid methyl ester. 1 ¹H NMR (400 MHz, chloroform-d) δ = 7.68–7.51 (m, 1H), 7.26–7.20 (m, 1H), 5.66–5.34 (m, 1H), 4.18–4.05 (m, 2H), 3.76 (s, 3H), 3.05–2.87 (m, 2H), 2.87–2.64 (m, 2H), 1.57 (s, 3H).

[0291] 2-Bromo-2-((S)-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-8-yl)acetate was prepared as a yellow solid (79.6% yield) starting from arbitrarily specified (S)-2-(6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-8-yl)acetate.

[0292] Methyl 2-bromo-2-((R)-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-8-yl)acetate is prepared starting from any specified (R)-2-(6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-8-yl)acetate using a procedure similar to that described in step 7 above.

[0293] Methyl 2-(1'-(2,2-dimethyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate was separated using a procedure similar to that described in step 6 above. (SFC column: DAICELCHIRALPAKIG (250 mm)) 30 mm, 10 μm); mobile phase: [CO2-MeOH]; B%: 20%, isocratic elution mode), to obtain arbitrarily specified (S)-2-(1'-(2,2-dimethyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)methyl acetate and arbitrarily specified (R)-2-(1'-(2,2-dimethyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)methyl acetate.

[0294] Methyl 2-bromo-2-(1'-((R)-2,2-dimethyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate as a yellow oil was prepared by a procedure similar to that described in step 7 above, starting with methyl (R)-2-(1'-((R)-2,2-dimethyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate. 1 H NMR (400 MHz, CDCl3) δ=7.38 (d, J =8.0Hz, 1H), 7.30-7.28 (m, 1H), 7.14 (d, J=8.1 Hz, 1H), 5.00 (s, 1H), 4.83-4.69 (m, 1H), 3.96-3.90 (m, 1H), 3.86-3.81 (m, 1H), 3.80 (s, 3H), 2.52-2.43(m, 1H), 2.35-2.28 (m, 1H), 1.98-1.91 (m, 1H), 1.78 (s, 2H), 1.71-1.60 (m,3H), 1.36 (s, 3H), 1.32 (s, 3H).

[0295] Methyl 2-bromo-2-(1'-((S)-2,2-dimethyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate is prepared starting from any specified (S)-2-(1'-((S)-2,2-dimethyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate using a procedure similar to that described in step 7 above.

[0296] Using a procedure similar to that described in step 6 above, via SFC (column: DAICEL CHIRALPAKIC (250 mm) 30 mm, 10 μm); mobile phase: [CO2-IPA (0.1% NH3H2O)]; B%: 25%, isocratic elution mode) to separate methyl (S)-2-(4-methylchroman-5-yl)acetate to give two compounds. Peak 1, arbitrarily designated (S)-2-(4-methylchroman-5-yl)acetate, was obtained as a yellow oil. SFC R t =0.900 min. 1 H NMR (400 MHz, CD3OD) δ=7.00 (t, J =7.9Hz, 1H), 6.73 (dd, J =0.8, 7.4Hz, 1H), 6.65 (d, J =8.3Hz, 1H), 4.23-4.15 (m, 2H), 3.75-3.57 (m, 5H), 3.14-3.01 (m, 1H), 2.16-1.94 (m, 1H), 1.75(qd, J =2.2, 13.8 Hz, 1H), 1.24 (d, J =7.0 Hz, 3H). Peak 2 was obtained as a yellow oil, arbitrarily specified as (R)-2-(4-methylchroman-5-yl)acetic acid methyl ester. SFC Rt =1.170 min. 1 H NMR (400 MHz, CD3OD) δ=7.00 (t, J =7.9Hz, 1H), 6.73 (dd, J =0.7, 7.4Hz, 1H), 6.66 (d, J =8.3Hz, 1H), 4.28-4.12 (m, 2H), 3.76-3.69 (m, 1H), 3.67 (s, 3H), 3.65-3.58 (m, 1H), 3.12-3.01(m, 1H), 2.11-1.95 (m, 1H), 1.82-1.67 (m, 1H), 1.24 (d, J=7.1 Hz, 3H).

[0297] 2-Bromo-2-((S)-4-methyl-5-yl)acetic acid methyl ester as a yellow oil was prepared from any specified (S)-2-(4-methyl-2-(4-methyl-5-yl)acetic acid methyl ester using a procedure similar to that described in step 7 above. LCMS [M+1] = 299.0. 1 H NMR (400 MHz, CDCl3) δ=7.33-7.21 (m, 1H), 7.14 (dt, J =2.7, 8.0Hz, 1H), 6.80 (ddd, J =0.8, 3.5, 8.0Hz, 1H), 5.67 (d, J =1.8 Hz, 1H),4.33-4.19 (m, 2H), 3.80 (d, J =19.9Hz, 3H), 3.26-3.10 (m, 1H), 2.21-2.08 (m,1H), 1.78 (qdd, J =2.2, 4.5, 13.8 Hz, 1H), 1.39 (dd, J =2.3, 7.1 Hz, 3H).

[0298] 2-Bromo-2-((R)-4-methyl-5-yl)acetic acid methyl ester as a yellow oil was prepared by a procedure similar to that described in step 7 above, starting with arbitrarily specified (R)-2-(4-methyl-2-(4-methyl-5-yl)acetic acid methyl ester. LCMS [M+1] = 219.1.

[0299] 2-Bromo-2-((S)-1-methylisochroman-8-yl)acetic acid methyl ester as a yellow oil was prepared from an arbitrarily selected (S)-2-(1-methylisochroman-8-yl)acetic acid methyl ester (92.03% yield) using a procedure similar to that described in step 7 above. LCMS [M+1] = 300.9.

[0300] Two compounds were prepared starting with 8-bromo-1-methylisochroman-8-yl) using a procedure similar to that described in steps 5 and 6 above. Separation conditions: (Column: ChiralPak IH, 250 × 50 mm, 10 μm; Mobile phase: [CO2-heptane:IPA = 1:1 (0.1% NH3H2O)]; B%: 11%, isocratic elution mode)). Peak 1, as a yellow oil, was arbitrarily designated as (R)-2-(1-methylisochroman-8-yl)acetate. Peak 2, also as a yellow oil, was arbitrarily designated as (S)-2-(1-methylisochroman-8-yl)acetate. 1 H NMR (400 MHz, CDCl3) δ=7.19-7.14 (m, 1H), 7.13-7.09 (m,1H), 7.05 (d, J =7.2 Hz, 1H), 5.11 (q, J =6.6Hz, 1H), 4.13-4.06 (m, 1H), 3.86-3.80 (m, 1H), 3.71 (s, 3H), 3.67-3.60 (m, 1H), 3.58-3.51 (m, 1H), 2.97-2.90(m, 1H), 2.84-2.76 (m, 1H), 1.49 (d, J =6.6Hz, 3H).

[0301] Separation was performed using a procedure similar to that described in steps 5, 6, and 7 above, from (3R,4S)-5-bromo-3,4,7-trimethylisocyanate (SFC). Separation conditions: Column: DAICL CHIRALPAK AD (250 mm). Two compounds were prepared starting at 50 mm, 10 μm; mobile phase: [CO2-i-PrOH (0.1% NH3·H2O )]; B%: 11%-11%, 2 min). Peak 1, as a yellow oil, was obtained and arbitrarily designated as 2-bromo-2-((3R,4S)-3,4,7-trimethylisochroman-5-yl)tert-butyl acetate. Peak 2, as a yellow oil, was obtained and arbitrarily designated as 2-bromo-2-((3S,4R)-3,4,7-trimethylisochroman-5-yl)tert-butyl acetate.

[0302] Separation was performed using a procedure similar to that described in steps 5, 6, and 7 above, from 5-bromo-7-fluoro-4-methylisocyanate (SFC). Separation conditions: Column: DAICL CHIRALPAK IC (250 mm). Two compounds were prepared starting with a mobile phase of [CO2-i-PrOH(0.1%NH3H2O)] and 18% B isocratic elution mode. Peak 1, as a yellow oil, was obtained and arbitrarily designated as methyl 2-bromo-2-((S)-7-fluoro-4-methylisochroman-5-yl)acetate. Peak 2, as a yellow oil, was obtained and arbitrarily designated as methyl 2-bromo-2-((R)-7-fluoro-4-methylisochroman-5-yl)acetate.

[0303] Preparation of (S)-2-(1-methylisochroman-8-yl)methyl acetate and (R)-2-(1-methylisochroman-8-yl)methyl acetate Step 1: 7-Bromo-1-methyl-2,3-dihydro-1H-inden-1-ol 7-Bromo-2,3-dihydro-1H-indanone (20 g, 94.76 mmol, 1 equivalent) was added to a solution of MeMgBr (3 M, 157.94 mL, 5 equivalents) in THF (200 mL) at 0 °C under N2. The mixture was stirred at 0 °C for 2 hours. TLC (petroleum ether / ethyl acetate = 5 / 1, R f = 0.35) indicates the formation of a new spot, and 7-bromo-2,3-dihydro-1H-inden-1-one is completely depleted. The reaction mixture was quenched with aqueous NH4Cl solution (500 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (400 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). 7-bromo-1-methyl-2,3-dihydro-1H-inden-1-ol was given as a yellow oil (88.29% yield). 1 H NMR (400 MHz, CDCl3) δ=7.35 (d, J=7.8Hz, 1H), 7.19-7.14 (m, 1H), 7.11-7.06 (m, 1H), 2.97-2.91 (m, 1H), 2.82 (t, J=8.8 Hz, 1H), 2.27 (dd, J=5.4, 8.7Hz, 2H), 1.64 (s, 3H).

[0304] Step 2: 4-Bromo-3-methyl-1H-indene TsOH (288.14 mg, 1.67 mmol, 0.02 equivalents) was added to a solution of 7-bromo-1-methyl-2,3-dihydro-1H-inden-1-ol (19 g, 83.66 mmol, 1 equivalent) in toluene (150 mL) at 20 °C. The mixture was stirred at 135 °C for 2 hours. TLC (petroleum ether / ethyl acetate = 3 / 1, R f = 0.85) indicates the formation of a new spot and complete depletion of 7-bromo-1-methyl-2,3-dihydro-1H-indene-1-ol. The reaction mixture was quenched with H2O (300 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (200 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). 4-Bromo-3-methyl-1H-indene was given as a colorless solid (97.18% yield). 1 H NMR (400 MHz, CDCl3) δ=7.47-7.35(m, 2H), 7.02 (t, J=7.6Hz, 1H), 6.33-6.20 (m, 1H), 3.30 (t, J=2.2 Hz, 2H), 2.43 (q, J=2.2 Hz, 3H).

[0305] Step 3: 2-(3-bromo-2-(1-hydroxyethyl)phenyl)ethanol-1-ol O3 (11.48 g, 239.14 mmol, 5 equivalents) was added to a solution of 4-bromo-3-methyl-1H-indene (10 g, 47.83 mmol, 1 equivalent) in DCM (100 mL) and MeOH (20 mL) at -70 °C. The mixture was stirred at -70 °C for 0.5 h. NaBH4 (2.35 g, 62.18 mmol, 1.3 equivalents) was added at -70 °C. The mixture was stirred at 15 °C for 2 h. TLC (petroleum ether / ethyl acetate = 5 / 1, R f= 0.25) indicates the formation of a new spot, and 4-bromo-3-methyl-1H-indene is completely depleted. The reaction mixture was quenched with NH4Cl (200 mL) and extracted with CH2Cl2 (100 mL × 3). The combined organic layers were washed with brine (200 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was used in the next step without purification. 2-(3-bromo-2-(1-hydroxyethyl)phenyl)ethyl-1-ol (6.4 g, crude substance) was given as a white solid.

[0306] Step 4: 8-Bromo-1-methylisochromosome H3PO4 (26.04 g, 265.73 mmol, 15.50 mL, 10.51 equivalents) was added to a solution of 2-(3-bromo-2-(1-hydroxyethyl)phenyl)ethanol-1-ol (6.2 g, 25.29 mmol, 1 equivalent) in CH3CN (60 mL) at 20 °C under N2. The mixture was stirred at 80 °C for 12 hours. TLC (petroleum ether / ethyl acetate = 3 / 1, R f =0.5) indicates the formation of a new spot, and 2-(3-bromo-2-(1-hydroxyethyl)phenyl)ethanol-1-ol remains. The reaction mixture was quenched with H2O (200 mL) and extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (200 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). 8-bromo-1-methylisochromanol was given as a colorless oil (52.23% yield).

[0307] Step 5: Methyl 2-(1-methylisochroman-8-yl)acetate TMSCl (2.13 g, 19.61 mmol, 2.49 mL, 0.1 equivalent) was added to a mixture of Zn (15.39 g, 235.33 mmol, 1.2 equivalent) in THF (196.11 mL) at 20 °C, and then stirred at 20 °C for 0.5 h. BrCH2COOt-Bu (30 g, 196.11 mmol, 18.52 mL, 1 equivalent) was added dropwise at 20 °C, and then stirred at 20 °C for 12 h. At 25°C under N2, 8-bromo-1-methylisocyanate (3 g, 13.21 mmol, 1 equivalent) in THF (25 mL) was added to a solution of (Pd t-Bu3P)2 (675.11 mg, 1.32 mmol, 0.1 equivalent) and the above mixture (1 M, 39.63 mL, 3 equivalent). The mixture was stirred at 80°C for 1 hour. TLC (petroleum ether / ethyl acetate = 2 / 1, R f =0.26) indicates that 8-bromo-1-methylisochroman-8-yl) is completely depleted and forms a new spot. The reaction mixture was filtered, and the filter cake was washed with ethyl acetate. The reaction mixture was then quenched by adding H2O (20 mL) at 20 °C and extracted with ethyl acetate (10 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). Methyl 2-(1-methylisochroman-8-yl)acetate was given as a white solid (68.73% yield). 1 H NMR (400 MHz, CDCl3) δ=7.19-7.13(m, 1H), 7.13-7.09 (m, 1H), 7.05 (d, J =7.4Hz, 1H), 5.11 (q, J =6.6Hz, 1H), 4.12-4.07 (m, 1H), 3.86-3.80 (m, 1H), 3.70 (s, 3H), 3.66-3.60 (m, 1H), 3.57-3.51(m, 1H), 3.00-2.89 (m, 1H), 2.82-2.73 (m, 1H), 1.49 (d, J =6.5Hz, 3H).

[0308] Step 6: (S)-2-(1-Methylisochroman-8-yl)methyl acetate and (R)-2-(1-Methylisochroman-8-yl)methyl acetate Through SFC (Column: DAICL CHIRALCEL OJ (250mm)) 50 mm, 10 μm); Mobile phase: [heptane-EtOH]; B%: 20%, isocratic elution mode) Separate methyl 2-(1-methylisochroman-8-yl)acetate to obtain two peaks (R t1 =2.93min and R t2 =3.44min).

[0309] Peak 1 (40.00% yield) was obtained as a yellow oil and arbitrarily designated as (S)-2-(1-methylisochroman-8-yl)acetic acid methyl ester. 1 ¹H NMR (400 MHz, chloroform-d) δ = 7.19–7.14 (m, ¹H), 7.13–7.09 (m, ¹H), 7.05 (d, J =7.3Hz, 1H), 5.11 (q, J =6.5Hz, 1H), 4.11 (dd, J =4.4, 8.6, 11.4Hz, 1H),3.87-3.80 (m, 1H), 3.71 (s, 3H), 3.66-3.60 (m, 1H), 3.58-3.51 (m, 1H), 2.99-2.90 (m, 1H), 2.83-2.75 (m, 1H), 1.49 (d, J =6.5Hz, 3H).

[0310] Peak 2 (40.00% yield) was obtained as a yellow oil and arbitrarily designated as (R)-2-(1-methylisochroman-8-yl)acetic acid methyl ester. 1 ¹H NMR (400 MHz, chloroform-d) δ = 7.19–7.14 (m, ¹H), 7.13–7.08 (m, ¹H), 7.05 (d, J =7.4Hz, 1H), 5.11 (q, J =6.5Hz, 1H), 4.11 (ddd, J =4.5, 8.6, 11.4Hz,1H), 3.83 (td, J =5.0, 11.3Hz, 1H), 3.71 (s, 3H), 3.66-3.61 (m, 1H), 3.58-3.50(m, 1H), 2.99-2.89 (m, 1H), 2.84-2.75 (m, 1H), 1.49 (d, J =6.5Hz, 3H).

[0311] Preparation of methyl 2-(6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-8-yl)acetate Step 1: 2-(2-acetyl-3-bromo-5-fluorophenyl)acetic acid A mixture of 1-(2-bromo-4-fluorophenyl)ethyl-1-one (8.50 g, 39.16 mmol, 1 equivalent), 5-diazonyl-2,2-dimethyl-1,3-dioxane-4,6-dione (9.99 g, 58.75 mmol, 1.5 equivalent), [Ir(Cp)Cl2]2 (315.97 mg, 391.64 μmol, 0.01 equivalent), AgNTf2 (258.33 mg, 665.79 μmol, 0.017 equivalent), and PhB(OH)2 (4.78 g, 39.16 mmol, 1 equivalent) in H2O (170 mL) was stirred at 80 °C for 16 h. LCMS showed 66% of the starting material remaining and approximately 16% of the desired mass was detected. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL × 2). The combined organic layers were washed with brine (20 mL × 2), dried over Na₂SO₄, and concentrated under reduced pressure to obtain the residue. The residue was purified by rapid silica gel chromatography (Biotage®; 40 g SepaFlash® silica gel column, eluent 0–10% ethyl acetate / petroleum ether gradient, @ 80 mL / min) to give 2-(2-acetyl-3-bromo-5-fluorophenyl)acetic acid (9.3% yield) as a white solid. LCMS: [M+1] = 275.0. 1 ¹H NMR (400 MHz, chloroform-d) δ=7.27 (dd, J=2.3, 7.8 Hz, 1H), 7.00 (dd, J=2.3, 8.8 Hz, 1H), 3.62 (s, 2H), 2.59 (s, 3H).

[0312] Step 2: 2-(2-acetyl-3-bromo-5-fluorophenyl)-N,N-dimethylacetamide N-methylmethylamine; N-methylmethylamine hydrochloride (1.30 g, 16.00 mmol, 2.3 equivalents), DIEA (3.15 g, 24.35 mmol, 4.24 mL, 3.5 equivalents), DMAP (1.44 g, 11.83 mmol, 1.7 equivalents), and HATU (4.76 g, 12.52 mmol, 1.8 equivalents) were added to a solution of 2-(2-acetyl-3-bromo-5-fluorophenyl)acetic acid (1.91 g, 6.96 mmol, 1 equivalent) in DCM (25 mL), and the mixture was stirred at 25 °C for 16 h. LCMS showed approximately 30.9% of the desired mass. Water (30 mL) was added, and the mixture was extracted with DCM (30 mL × 2). The combined organic layers were washed with brine (30 mL × 2), dried over Na₂SO₄, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, 20 g SepaFlash® silica gel rapid column, eluent 0–30% petroleum ether gradient / ethyl acetate, @ 120 mL / min) to give the product 2-(2-acetyl-3-bromo-5-fluorophenyl)-N,N-dimethylacetamide (66.6% yield) as a yellow oil. LCMS: [M+1] = 302.0.

[0313] Step 3: 2-(3-bromo-5-fluoro-2-(1-hydroxyethyl)phenyl)-N,N-dimethylacetamide LiBH4 (2 M, 4.63 mL, 1 equivalent) was added to a solution of 2-(2-acetyl-3-bromo-5-fluorophenyl)-N,N-dimethylacetamide (1.4 g, 4.63 mL, 1 equivalent) in THF (20 mL) at 0 °C, and the mixture was stirred at 25 °C for 12 hours. LCMS showed approximately 66% of the desired mass. The mixture was poured into ice water (20 mL) and extracted with ethyl acetate (20 mL × 2). The combined organic layers were washed with brine (20 mL × 2), dried over Na2SO4, and concentrated under reduced pressure to give a residue to 2-(3-bromo-5-fluoro-2-(1-hydroxyethyl)phenyl)-N,N-dimethylacetamide (1.1 g, crude) as a yellow oil. LCMS: [M+1] = 286.0.

[0314] Step 4: 8-Bromo-6-fluoro-1-methylisochroman-3-one HCl (3 M, 1.21 mL, 1 equivalent) was added to a solution of 2-(3-bromo-5-fluoro-2-(1-hydroxyethyl)phenyl)-N,N-dimethylacetamide (1.1 g, 3.62 mmol, 1 equivalent) in THF (12 mL), and the mixture was stirred at 90 °C for 2 h. LCMS showed approximately 54% of the desired mass. Water (15 mL) was added, and the mixture was extracted with ethyl acetate (15 mL × 2). The combined organic layers were washed with brine (15 mL × 2), dried over Na₂SO₄, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO₂, 20 g SepaFlash® silica gel rapid column, eluent 0–30% petroleum ether gradient / ethyl acetate, @ 120 mL / min) to give 8-bromo-6-fluoro-1-methylisochroman-3-one as a yellow oil (59.7% yield). LCMS :[M+1]=259.0. 1 ¹H NMR (400 MHz, chloroform-d) δ=7.20–7.18 (m, 1H), 6.84 (brd, J=8.1 Hz, 1H), 5.73 (q, J=6.9 Hz, 1H), 3.71–3.61 (m, 2H), 1.55 (d, J=6.9 Hz, 3H).

[0315] Step 5: 8-Bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-3-one 8-Bromo-6-fluoro-1-methylisocyano-3-one (1.7 g, 6.56 mmol, 1 equivalent) in DMF (10 mL) was added to a solution of Cs₂CO₃ (6.41 g, 19.69 mmol, 3 equivalents) in DMF (10 mL) at 20 °C under N₂. The mixture was stirred at 20 °C for 0.5 h. 1-Bromo-2-(2-bromoethoxy)ethane (6.09 g, 26.25 mmol, 3.30 mL, 4 equivalents) was added at 20 °C under N₂. The mixture was stirred at 20 °C for 12 h. LCMS showed a peak with the desired mass. The reaction mixture was quenched with H₂O (100 mL) and extracted with ethyl acetate (30 mL × 3). The combined organic layers were washed with brine (100 mL × 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1) to give 8-bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro-[isochroman-4,4'-pyran]-3-one as a colorless oil (69.4% yield). LCMS: [M+1] = 329.1. 1 ¹H NMR (400 MHz, chloroform-d) δ=7.31 (dd, J =2.4, 7.5Hz, 1H), 7.24 (dd, J =2.3, 10.1 Hz, 1H), 5.76 (q, J =6.6Hz, 1H), 4.50 (dt, J =2.5, 11.6Hz, 1H), 4.05-3.86(m, 3H), 2.34 (ddd, J =5.6, 11.6, 13.9Hz, 1H), 2.04-1.90 (m, 3H), 1.72 (d, J =6.8Hz, 3H).

[0316] Step 6: 8-Bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-3-ol DIBAL-H (1 M, 5.92 mL, 1.3 equivalents) was added to a solution of 8-bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-3-one (1.5 g, 4.56 mmol, 1 equivalent) in CH2Cl2 (20 mL) at 0 °C under N2. The mixture was stirred at 0 °C for 1 h. TLC (petroleum ether / ethyl acetate = 3 / 1, Rf = 0.25) indicated the formation of a new spot. An aqueous solution of HCl (1 M, 50 mL) was added to the reaction mixture, which was stirred for 30 min and then extracted with DCM (30 mL × 3). The combined organic layers were washed with brine (100 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1) to give 8-bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-3-ol (800 mg, 2.42 mmol, 53.0%) as a white solid. 1 ¹H NMR (400 MHz, chloroform-d) δ=7.25-7.15 (m, 2H), 5.77-5.46 (m, 1H), 5.11 (q, J=6.4Hz, 1H), 4.01-3.77 (m, 4H), 2.29-2.06 (m, 2H), 1.82-1.59 (m, 5H).

[0317] Step 7: 8-Bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran] Et3SiH (1.47 g, 12.68 mmol, 2.03 mL, 6 equivalents) and TFA (2.17 g, 19.02 mmol, 1.41 mL, 9 equivalents) were added to a solution of 8-bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-3-ol (700 mg, 2.11 mmol, 1 equivalent) in DCM (14 mL) at 20 °C under N2. The mixture was stirred at 50 °C for 24 h. TLC (plate 1, petroleum ether / ethyl acetate = 3 / 1, Rf = 0.5) indicated the formation of a new spot. The reaction mixture was quenched with H2O (30 mL) and extracted with DCM (10 mL × 3). The combined organic layers were washed with brine (20 mL × 2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1) to give 8-bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran] (1.1 g, crude matter) as a white solid. 1 H NMR (400 MHz, chloroform-d) δ=7.20-7.13 (m, 2H), 5.04 (q, J=6.4Hz, 1H), 4.10 (d, J=12.1 Hz, 1H), 3.97-3.87 (m, 3H), 3.75-3.67(m, 1H), 3.58 (dt, J=2.1, 12.4Hz, 1H), 2.23 (dt, J=5.1, 13.3Hz, 1H), 1.90-1.81 (m, 1H), 1.77-1.70 (m, 1H), 1.60 (d, J=6.4Hz, 3H), 1.56 (brd, J=4.8 Hz,1H).

[0318] Step 8: Methyl 2-(6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-8-yl)acetate Pd(t-Bu3P)2 (162.15 mg, 317.28 μmol, 0.1 equivalent) was added to a mixture of 8-bromo-6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran] (1.0 g, 3.17 mmol, 1 equivalent) in THF (5 mL) at 25 °C. BrZnCH2COOMe (1 M, 6.98 mL, 2.2 equivalent) was added to the reactants. The reactants were stirred at 80 °C for 3 hours. TLC (petroleum ether:ethyl acetate = 2:1) showed the starting material (R... f = 0.50) was consumed and a new point (R) was observed. f = 0.30). Filter the mixture and add H2O to the filtrate. Filter the mixture and separate the layers, extracting the aqueous phase with EtOAc (20 mL × 3). The combined organic layers were washed with brine (20 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by rapid silica gel chromatography (petroleum ether / ethyl acetate = 2 / 1). The product was further purified by SFC (conditions: column: DAICEL CHIRALPAK AD (250 mm)). (30mm, 10μm); Mobile phase: [CO2-EtOH]; B%: 15%, isocratic elution mode) separation.

[0319] Peak 1 (15.3% yield) was obtained as a yellow oil and arbitrarily designated as (S)-2-(6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-8-yl)acetic acid methyl ester. 1 H NMR (400 MHz, chloroform-d) δ=7.10(dd, J=2.6, 10.3Hz, 1H), 6.87 (dd, J=2.6, 9.1 Hz, 1H), 5.07 (q, J=6.5Hz, 1H),4.06-3.95 (m, 2H), 3.93-3.83 (m, 2H), 3.73 (s, 3H), 3.75-3.66 (m, 1H), 3.64-3.48 (m, 3H), 2.21 (dt, J=5.1, 13.3Hz, 1H), 1.92 (dt, J=5.1, 13.3Hz, 1H),1.76-1.50 (m, 4H), 1.49 (d, J = 6.5 Hz, 3H).

[0320] Peak 2 (15.3% yield) was obtained as a yellow oil and arbitrarily designated as (R)-2-(6-fluoro-1-methyl-2',3',5',6'-tetrahydrospiro[isochroman-4,4'-pyran]-8-yl)acetic acid methyl ester. 1 H NMR (400 MHz, chloroform-d) δ=7.09(dd, J=2.6, 10.3Hz, 1H), 6.87 (dd, J=2.5, 9.1 Hz, 1H), 5.07 (q, J=6.5Hz, 1H), 4.00 (q, J=12.0Hz, 2H), 3.95-3.84 (m, 2H), 3.80-3.67 (m, 4H), 3.64-3.47 (m,3H), 2.21 (dt, J=5.2, 13.4Hz, 1H), 1.92 (dt, J=5.1, 13.2 Hz, 1H), 1.74-1.59 (m, 2H), 1.48 (d, J=6.5Hz, 3H).

[0321] Separation of isomers of methyl 2-(6'-methyl-1'-(2-methyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate 2-(6'-methyl-1'-(2-methyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)methyl acetate (4.8 g, 16.15 mmol, 1 equivalent) was analyzed by SFC (column: DAICEL CHIRALPAK IC (250 mm)). 50 mm, 10 μm); Mobile phase: [CO2-MeOH (0.1%NH3H2O)]; B%: 35%, isocratic elution mode and column: REGIS (s,s) WHELK-O1 (250 mm) Separation was performed using a mobile phase of [CO2-IPA] (50 mm, 10 μm) and a B% elution mode (25% B, isocratic elution mode) to obtain four isomers.

[0322] Peak 1, obtained as a yellow oil, was arbitrarily designated as methyl 2-(6'-methyl-1'-((2S, 4S)-2-methyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (13.00% yield). SFC: Rt = 2.228 min. 1¹H NMR (400 MHz, chloroform-d) δ=6.92 (s, ¹H), 6.74 (s, ¹H), 4.66–4.45 (m, ¹H), 4.15 (dd, ¹H) J =3.8, 11.9Hz, 1H), 3.71 (s, 3H), 3.65-3.52 (m, 2H), 3.35(s, 2H), 2.50 (dq, J =4.7, 12.6Hz, 1H), 2.38 (s, 3H), 2.25-2.11 (m, 1H), 1.89-1.85 (m, 2H), 1.75 (td, J =2.0, 12.6Hz, 1H), 1.68 (brd, J =2.4Hz, 1H), 1.61 (brs,2H), 1.28 (d, J =6.2 Hz, 3H).

[0323] Peak 2, obtained as a yellow oil, was arbitrarily designated as methyl 2-(6'-methyl-1'-((2R, 4R)-2-methyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (13.00% yield). SFC: Rt = 2.450 min. 1 ¹H NMR (400 MHz, chloroform-d) δ=6.92 (s, ¹H), 6.74 (s, ¹H), 4.57 (tdd, J =4.3, 8.3, 12.6Hz, 1H), 4.15 (dd, J =4.5, 11.1 Hz, 1H), 3.71 (s, 3H), 3.65-3.55(m, 2H), 3.35 (s, 2H), 2.50 (dq, J =4.8, 12.6Hz, 1H), 2.38 (s, 3H), 2.26-2.11(m, 1H), 1.91-1.82 (m, 2H), 1.75 (td, J =2.0, 12.7Hz, 1H), 1.68 (brd, J =2.0Hz,1H), 1.65-1.61 (m, 2H), 1.28 (d, J =6.2 Hz, 3H).

[0324] Peak 3, obtained as a yellow oil, was arbitrarily designated as methyl 2-(6'-methyl-1'-((2R, 4S)-2-methyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (31.7% yield). SFC: Rt = 2.794 min. 1 ¹H NMR (400 MHz, chloroform-d) δ=6.91 (s, 1H), 6.75 (s, 1H), 4.79-4.68 (m, 1H), 4.40-4.32 (m, 1H), 3.92-3.86 (m, 2H), 3.71 (s, 3H), 3.35 (s, 2H), 2.70-2.61 (m, 1H), 2.56-2.45 (m, 1H), 2.38 (s, 3H), 1.89-1.83 (m, 2H), 1.77-1.70 (m, 1H), 1.65-1.57 (m, 3H), 1.39 (d, J =6.9Hz, 3H).

[0325] Peak 4, obtained as a yellow oil, was arbitrarily designated as methyl 2-(6'-methyl-1'-((2S, 4R)-2-methyltetrahydro-2H-pyran-4-yl)-2'-oxospiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (33.33% yield). SFC: Rt = 3.106 min. 1 ¹H NMR (400 MHz, chloroform-d) δ=6.91 (s, 1H), 6.75 (s, 1H), 4.87-4.61 (m, 1H), 4.44-4.28 (m, 1H), 3.93-3.82 (m, 2H), 3.71 (s, 3H), 3.35 (s, 2H), 2.70-2.60 (m, 1H), 2.56-2.44 (m, 1H), 2.38 (s, 3H), 1.88-1.82 (m, 2H), 1.77-1.68 (m, 1H), 1.65-1.57 (m, 3H), 1.39 (d, J =6.8 Hz, 3H).

[0326] Preparation of methyl 2-bromo-2-(7'-fluoro-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate Step 1: Methyl 2-(6-bromo-2,3-difluorophenyl)-2-oxoacetate LDA (2 M, 25.91 mL, 1 equivalent) was added dropwise to a solution of 4-bromo-1,2-difluorobenzene (10 g, 51.82 mmol, 1 equivalent) in THF (100 mL) at -70 °C under a nitrogen atmosphere. The reaction mixture was allowed to stir at -70 °C for 0.5 h. A solution of dimethyl oxalate (6.12 g, 51.82 mmol, 1 equivalent) in THF (50 mL) was added to the mixture at -70 °C. The reaction mixture was allowed to stir at -70 °C for 1 h. The reaction mixture was quenched by adding an aqueous solution of NH4Cl (200 mL) and extracted with ethyl acetate (200 mL × 2). The organic solutions were combined, washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give the residue. 2-(6-bromo-2,3-difluorophenyl)-2-oxoacetic acid methyl ester (8.7 g, crude matter) was obtained as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ=7.40 (ddd, J =1.8,3.9, 8.9Hz, 1H), 7.27-7.20 (m, 1H), 3.98 (s, 3H).

[0327] Step 2: 2-(6-bromo-2,3-difluorophenyl)-2-oxo-N-(tetrahydro-2H-pyran-4-yl)acetamide Tetrahydro-2H-pyran-4-amine (4.73 g, 46.77 mmol, 1.5 equivalents) was added to methyl 2-(6-bromo-2,3-difluorophenyl)-2-oxoacetate (8.7 g, 31.18 mmol, 1 equivalent) in MeOH (100 mL) under a nitrogen atmosphere at 25 °C. The reaction mixture was allowed to be stirred at 25 °C under a nitrogen atmosphere for 16 hours. The mixture was concentrated under reduced pressure. The crude product was milled with petroleum ether / ethyl acetate (5 / 1). 2-(6-bromo-2,3-difluorophenyl)-2-oxo-N-(tetrahydro-2H-pyran-4-yl)acetamide (73.70% yield) was given as a yellow oil. LCMS [M+1] = 348.0. 1 H NMR (400 MHz, CDCl3) δ=7.36 (ddd, J =1.8, 3.9, 8.9Hz, 1H), 7.26-7.14 (m, 1H), 6.88 (brd, J=6.4Hz, 1H), 4.05-3.94 (m, 3H), 3.56-3.49 (m, 2H), 2.05-1.92 (m, 2H), 1.67-1.58 (m, 2H).

[0328] Step 3: 4-Bromo-7-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione Potassium 2-methylbutan-2-olate (1 M, 34.47 mL, 1.5 equivalent) was added to a solution of 2-(6-bromo-2,3-difluorophenyl)-2-oxo-N-(tetrahydro-2H-pyran-4-yl)acetamide (8 g, 22.98 mmol, 1 equivalent) in toluene (100 mL) at 0 °C under a N2 atmosphere. The mixture was allowed to be stirred at 25 °C for 1 hour. The reaction mixture was quenched by adding cold aqueous HCl (1 M, 100 mL) and extracted with ethyl acetate (100 mL × 2). The organic solutions were combined, washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give 4-bromo-7-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione (66.31% yield) as a yellow solid. LCMS[M+1]=328.0. 1 H NMR (400 MHz, CDCl3) δ=7.25 (s, 1H), 7.23 (d, J=2.5Hz, 1H), 4.63-4.54 (m, 1H), 4.13 (dd, J=4.8, 11.7Hz, 2H), 3.54-3.49 (m, 2H), 2.55-2.46(m, 2H), 1.75 (brdd, J=2.4, 12.3Hz, 2H).

[0329] Step 4: 4-Bromo-7-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2-one Two batches were performed in parallel. TiCl4 (2.60 g, 13.71 mmol, 4.68 equivalents) was added dropwise to a mixture of Zn (1.4 g, 21.41 mmol, 4.68 equivalents) in THF (20 mL) at 0 °C under a N2 atmosphere. The mixture was allowed to stir at 80 °C for 2 hours. A solution of 4-bromo-7-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione (1.5 g, 4.57 mmol, 1 equivalent) in THF (20 mL) was added at 20 °C, and the mixture was allowed to stir at 20 °C for 2 hours. The reaction mixture was quenched by adding 100 mL of saturated aqueous NaHCO3 solution. The mixture was filtered and the filtrate was extracted with ethyl acetate (100 mL × 3). The organic solutions were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1). 4-Bromo-7-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2-one (69.63% yield) was obtained as a yellow oil from the combined batches. LCMS [M+1] = 314.0. 1 H NMR (400 MHz, CDCl3) δ=7.13 (dd, J =3.7, 8.9Hz, 1H), 6.97 (dd, J =8.9,11.8 Hz, 1H), 4.73-4.62 (m, 1H), 4.11 (dd, J =4.9, 11.6Hz, 2H), 3.55-3.48 (m,4H), 2.50-2.39 (m, 2H), 1.66 (brdd, J =2.4, 12.3Hz, 2H).

[0330] Step 5: 4'-Bromo-7'-Fluoro-1'-(Tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one Cs₂CO₃ (6.53 g, 20.05 mmol, 3 equivalents) and 1,2-dibromoethane (6.28 g, 33.42 mmol, 2.52 mL, 5 equivalents) were added to a solution of 4-bromo-7-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2-one (2.1 g, 6.68 mmol, 1 equivalent) in DMF (30 mL). The mixture was allowed to be stirred at 35 °C for 12 h. The reactants were quenched by adding H₂O (100 mL) at 0 °C and extracted with ethyl acetate (50 mL × 3). The organic solutions were combined, washed with brine (50 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residues were purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1). 4'-bromo-7'-fluoro-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one was given as a yellow solid (87.95% yield). LCMS [M+1] = 340.0. 1 H NMR (400 MHz, CDCl3) δ=7.07(dd, J=4.0, 8.9Hz, 1H), 6.90 (dd, J=8.9, 11.8 Hz, 1H), 4.81-4.69 (m, 1H), 4.11 (dd, J=4.8, 11.5Hz, 2H), 3.52 (brt, J=11.9Hz, 2H), 2.58-2.46 (m, 2H), 2.41 (q, J=3.9Hz, 2H), 1.72-1.65 (m, 2H), 1.61-1.58 (m, 2H).

[0331] Step 6: Methyl 2-(7'-fluoro-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate Pd(t-Bu3P)2 (90.14 mg, 176.37 μmol, 0.05 equivalent) and BrZnCH2COOMe (1 M, 10.58 mL, 3 equivalent) were added to a solution of 4'-bromo-7'-fluoro-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one (1.2 g, 3.53 mmol, 1 equivalent) in THF (10 mL) at 20 °C, and the reaction mixture was allowed to be stirred at 80 °C under a N2 atmosphere for 2 hours. The reaction mixture was quenched by adding H2O (30 mL) at 0 °C and filtered. The filtrate was extracted with ethyl acetate (30 mL × 3). The organic solutions were combined, washed with brine (30 mL × 3), dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1). Methyl 2-(7'-fluoro-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate was given as a yellow oil (19.56% yield). LCMS Rt = 0.445 min, [M+1] = 334.3. 1 H NMR (400 MHz, CDCl3) δ=6.99 (dd, J=8.6,12.0Hz, 1H), 6.94-6.86 (m, 1H), 4.86-4.63 (m, 1H), 4.13-4.08 (m, 2H), 3.71(s, 3H), 3.56-3.50 (m, 2H), 3.35 (s, 2H), 2.60-2.46 (m, 2H), 2.00-1.89 (m,2H), 1.72-1.65 (m, 4H).

[0332] Step 7: Methyl 2-bromo-2-(7'-fluoro-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate LDA (2 M, 269.98 μL, 1 equivalent) was added to a solution of methyl 2-(7'-fluoro-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate (180 mg, 539.97 μmol, 1 equivalent) in THF (5 mL) at -70 °C under a N2 atmosphere. After 0.5 hours, TMSCl (70.40 mg, 647.96 μmol, 82.24 μL, 1.2 equivalent) was added, and the mixture was allowed to stir at -70 °C for 0.5 hours. NBS (288.32 mg, 1.62 mmol, 3 equivalent) was added to THF (5 mL). The mixture was allowed to stir at -70 °C for 0.5 hours. The reactants were quenched with 30 mL of saturated aqueous NH4Cl solution and extracted with ethyl acetate (30 mL × 2). The organic solutions were combined, washed with 10 mL of brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 3 / 1). Methyl 2-bromo-2-(7'-fluoro-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate was given as a yellow oil (8.40% yield). LCMS [M+1] = 412.0. 1 H NMR (400 MHz, CDCl3) δ=7.40 (dd, J=4.2, 8.9Hz, 1H), 7.07 (dd, J=8.9, 11.9Hz, 1H), 4.95 (s,1H), 4.77-4.72 (m, 1H), 4.11-4.08 (m, 2H), 3.80 (s, 3H), 3.52 (s, 2H), 2.52 (brd, J=7.9Hz, 2H), 2.02-1.91 (m, 2H), 1.73-1.67 (m, 4H).

[0333] Following a procedure similar to steps 4, 5, 6, and 7 above for the synthesis of methyl 2-bromo-2-(6'-fluoro-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate, methyl 2-bromo-2-(6'-fluoro-2'-oxo-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-4'-yl)acetate was prepared from 4-bromo-6-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione. 1H NMR (400 MHz, CDCl3) δ=7.11 (dd, J=2.3,10.3Hz, 1H), 6.89 (dd, J=2.1, 9.0Hz, 1H), 4.94 (d, J=1.4Hz, 1H), 4.51 (tt, J=4.4, 12.6Hz, 1H), 4.13 (d, J=7.1 Hz, 2H), 3.81 (s, 3H), 3.53 (dt, J=1.8,12.0Hz, 2H), 2.56-2.45 (m, 2H), 1.78 (brd, J=2.3Hz, 2H), 1.72-1.67 (m, 2H),1.29-1.25 (m, 2H).

[0334] Preparation of 4-bromo-6-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione Step 1: Methyl 2-(2-bromo-4,6-difluorophenyl)-2-oxoacetate n-BuLi (2.5 M, 24.46 mL, 0.75 equivalents) was added dropwise to a solution of 1-bromo-3,5-difluoro-2-iodobenzene (26 g, 81.53 mmol, 1 equivalent) in toluene (400 mL) at -70 °C under a nitrogen atmosphere. The reaction mixture was allowed to stir at -70 °C for 0.5 h. Then, a solution of dimethyl oxalate (19.26 g, 163.07 mmol, 2 equivalents) in toluene (500 mL) was added to the reaction mixture at -70 °C. The reaction mixture was allowed to stir at -70 °C for 1 h. The reactants were quenched by adding an aqueous solution of NH4Cl (400 mL) and the mixture was extracted with ethyl acetate (100 mL × 2). The organic solutions were combined, washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 2 / 1). Three batches were conducted in parallel. Methyl 2-(2-bromo-4,6-difluorophenyl)-2-oxoacetate was obtained as a yellow oil (70.33% yield). 1 H NMR (400 MHz, CDCl3) δ=7.25 (td, J=1.9, 7.8 Hz, 1H), 6.92 (ddd, J=2.3, 8.4, 9.6Hz, 1H), 3.96 (s, 3H).

[0335] Step 2: 4-Bromo-6-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione n-BuLi (2.5 M, 14.34 mL, 1 equivalent) was added to a solution of tetrahydro-2H-pyran-4-amine (3.99 g, 39.42 mmol, 1.1 equivalent) in THF (50 mL) at 0 °C under a N2 atmosphere. The reaction mixture was allowed to be stirred at 0 °C for 0.5 h. The reaction mixture was then added to a solution of methyl 2-(2-bromo-4,6-difluorophenyl)-2-oxoacetate (10 g, 35.84 mmol, 1 equivalent) in THF (50 mL) at 0 °C under a N2 atmosphere. The mixture was allowed to be stirred at 25 °C for 0.5 h. The reaction mixture was quenched with 2 M HCl aqueous solution (100 mL) and extracted with ethyl acetate (100 mL × 2). The organic solutions were combined, washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was ground with petroleum ether / ethyl acetate (3 / 1). 4-Bromo-6-fluoro-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione was obtained as a yellow solid (42.52% yield). 1 H NMR (400 MHz, CDCl3) δ=7.00 (dd, J =1.2, 8.6Hz, 1H), 6.83 (dd, J =1.2, 8.9Hz, 1H), 4.36 (tt, J =4.3, 12.3Hz, 1H), 4.15 (brdd, J =4.4, 11.7Hz, 2H), 3.52 (brt, J =11.5Hz, 2H), 2.45 (dq, J =4.6, 12.5Hz, 2H), 1.74 (brdd, J =2.2, 12.5 Hz, 2H).

[0336] Preparation of methyl 2-bromo-2-((R)-4-methylisochroman-5-yl)acetate Step 1: Methyl 2-(2-bromophenyl)propionate LiHMDS (1 M, 240.10 mL, 1.1 equivalents) was added dropwise to a solution of methyl 2-(2-bromophenyl)acetate (50 g, 218.27 mmol, 1 equivalent) in THF (100 mL) at 0 °C under a N2 atmosphere, and the reaction mixture was allowed to stir at 0 °C for 1 h. MeI (30.98 g, 218.27 mmol, 13.59 mL, 1 equivalent) was added dropwise at 0 °C under a N2 atmosphere. The mixture was allowed to stir at 20 °C for 1 h. The reaction mixture was quenched by the addition of NH4Cl (500 mL) and extracted with ethyl acetate (200 mL × 3). The organic solutions were combined, washed with brine (100 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. Methyl 2-(2-bromophenyl)propionate (60 g, crude product) was obtained as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ=7.57 (d, J=8.2 Hz, 1H), 7.35-7.27 (m, 2H), 7.16-7.10 (m, 1H), 4.24 (q, J=7.2Hz, 1H), 3.69 (s, 3H), 1.50 (d, J=7.2 Hz, 3H).

[0337] Step 2: 2-(2-bromophenyl)prop-1-ol Two reactions were carried out in parallel. A solution of methyl 2-(2-bromophenyl-)propionate (30 g, 123.41 mmol, 1 equivalent) in THF (100 mL) was added to a solution of LiBH4 (8.04 g, 368.86 mmol, 2.99 equivalents) in THF (100 mL) at 0 °C. The mixture was allowed to be stirred at 20 °C for 12 h. The mixture was diluted with H2O (500 mL) and extracted with ethyl acetate (200 mL × 3). The organic solutions were combined, washed with brine (200 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). 2-(2-bromophenyl)prop-1-ol (37 g, 172.02 mmol, 78.72% yield after two steps) was used as a colorless oil. LCMS Rt=0.869, [M-18+1]=197.1. 1H NMR (400 MHz, CDCl3) δ=7.56 (d, J=7.9Hz, 1H), 7.32-7.27 (m,1H), 7.26 (d, J=6.8 Hz, 1H), 7.10-7.04 (m, 1H), 3.82-3.75 (m, 1H), 3.72-3.66 (m, 1H), 3.54-3.46 (m, 1H), 1.28 (d, J=7.0Hz, 3H).

[0338] Step 3: 5-Bromo-4-methylisochromosome 2-(2-bromophenyl)prop-1-ol (17 g, 79.04 mmol, 1 equivalent) and (HCHO) were added to TFA (100 mL). n A solution of (7.83 g, 86.94 mmol, 1.1 equivalents) was stirred at 100 °C for 16 h. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H₂O (200 mL) and extracted with ethyl acetate (100 mL × 3). The organic solutions were combined, washed with brine (200 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The mixture was analyzed by preparative HPLC (column: Phenomenex luna C18 250). 150mm Further purification was performed at 15 μm; mobile phase: [water (TFA)-ACN]; B%: 30%-60%, 20 min. 5-Bromo-4-methylisochromanol was obtained as a yellow oil (44.57% yield). LCMS Rt=1.031, [M-18+1]=209.0. 1 H NMR (400 MHz, CDCl3) δ=7.43 (d, J=7.9Hz, 1H), 7.05 (t, J=7.8 Hz,1H), 6.98-6.92 (m, 1H), 4.87-4.79 (m, 1H), 4.76-4.67 (m, 1H), 4.00-3.93 (m,1H), 3.80 (dd, J=2.8, 11.3Hz, 1H), 3.02-2.92 (m, 1H), 1.39 (d, J=6.9Hz, 3H).

[0339] Step 4: (R)-2-(4-methylisochroman-5-yl)methyl acetate and (S)-2-(4-methylisochroman-5-yl)methyl acetate BrZnCH2COOt-Bu (1.0 M, 60 mL, 2.27 equivalents) was added to a solution of 5-bromo-4-methylisochloromethane (6 g, 26.42 mmol, 1 equivalent) and Pd(t-Bu3P)2 (1.35 g, 2.64 mmol, 0.1 equivalent) in THF (60 mL) at 20 °C under a nitrogen atmosphere. The mixture was allowed to stir at 80 °C for 2 hours. The reaction mixture was diluted with H2O (200 mL) and filtered, then extracted with ethyl acetate (100 mL × 2). The organic solutions were combined, washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was poured into HCl / MeOH (100 mL) at 20 °C. The mixture was allowed to stir at 50 °C for 12 hours. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (50 mL) and extracted with ethyl acetate (20 mL × 3). The organic solutions were combined, washed with brine (20 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). A racemic mixture (5 g) was obtained as a yellow oil. The racemic mixture was purified by SFC (column: DAICEL CHIRALPAK AD (250 mm)). 50 mm, 10 μm); Mobile phase: [0.1% NH3H2O ​​IPA]; B%: 11%-11%, 1.6 min) separation.

[0340] Peak 1, obtained as a colorless oil, was arbitrarily designated as (S)-2-(4-methylisochroman-5-yl)acetic acid methyl ester (30.93% yield). LCMS [M-18+1] = 203.1. SFC Rt = 1.134. 1 H NMR (400 MHz, CDCl3) δ=7.20-7.12 (m, 2H), 6.95-6.89 (m, 1H), 4.89-4.74 (m, 2H), 3.97-3.91 (m, 1H), 3.85-3.80 (m, 1H), 3.76-3.72 (m, 1H), 3.71 (s, 3H), 3.65-3.60 (m, 1H), 2.93-2.85(m, 1H), 1.33 (d, J=7.0Hz, 3H).

[0341] Peak 2, obtained as a colorless oil, was arbitrarily designated as (R)-2-(4-methylisochroman-5-yl)acetic acid methyl ester (25.78% yield). LCMS [M-18+1]=203.1. SFC Rt=1.330. 1H NMR (400 MHz, CDCl3) δ=7.18-7.11 (m, 2H), 6.94-6.89 (m, 1H), 4.89-4.73 (m, 2H), 3.97-3.91 (m, 1H), 3.87-3.79 (m, 1H), 3.76-3.72 (m, 1H), 3.71 (s, 3H), 3.66-3.59 (m, 1H), 2.94-2.84(m, 1H), 1.33 (d, J=7.0Hz, 3H).

[0342] Step 5: Methyl 2-bromo-2-((S)-4-methylisochroman-5-yl)acetate LDA (2 M, 1.70 mL, 1.5 equivalents) was added to a solution of (S)-2-(4-methylisocyanuric-5-yl)acetate (500.00 mg, 2.27 mmol, 1 equivalent) in THF (10 mL) at -60 °C, and the mixture was stirred at -60 °C for 30 min. TMSCl (394.58 mg, 3.63 mmol, 460.96 μL, 1.6 equivalents) was added at -60 °C, and the mixture was stirred at -60 °C for 0.5 h. A solution of NBS (484.82 mg, 2.72 mmol, 1.2 equivalents) in THF (5 mL) was added at -60 °C. The mixture was stirred at -60 °C for 1 h. The residue was diluted with NH4Cl (20 mL) and extracted with ethyl acetate (10 mL × 3). The organic solutions were combined, washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 20 / 1). Methyl 2-bromo-2-((S)-4-methylisochroman-5-yl)acetate was given as a yellow oil (29.45% yield).

[0343] 2-bromo-2-((S)-4-methylisochroman-5-yl)acetic acid methyl ester was prepared by a similar procedure, starting from any specified (R)-2-(4-methylisochroman-5-yl)acetic acid methyl ester.

[0344] Preparation of methyl 2-((methylsulfonyl)oxy)-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetate Step 1: 4-(3-bromo-2-nitrophenoxy)tetrahydro-2H-pyran DIAD (18.09 g, 89.45 mmol, 17.34 mL, 1.5 equivalent), tetrahydro-2H-pyran-4-ol (7.31 g, 71.56 mmol, 7.15 mL, 1.2 equivalent), and PPh3 (18.77 g, 71.56 mmol, 1.2 equivalent) were added to a solution of 3-bromo-2-nitrophenol (13 g, 59.63 mmol, 1.2 equivalent) in toluene (200 mL). The mixture was allowed to be stirred at 100 °C for 2 h. The reactants were quenched by adding H2O (20 mL) at 20 °C and extracted with ethyl acetate (10 mL × 3). The organic solutions were combined, washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residues were purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). 4-(3-bromo-2-nitrophenoxy)tetrahydro-2H-pyran was obtained as a colorless oil (23 g, 53.29 mmol, 89.37% yield, 70% purity). 1 H NMR (400 MHz, CDCl3) δ=7.30-7.27 (m, 1H), 7.25-7.20 (m, 1H), 7.00 (dd, J =1.3, 8.3Hz, 1H), 4.64 (tt, J =3.5, 7.0Hz, 1H), 3.90 (ddd, J =3.5, 7.6, 11.5Hz, 2H), 3.64-3.56 (m, 2H), 2.05-1.99 (m, 2H), 1.83 (ddd, J =3.6, 6.9, 13.5Hz, 2H).

[0345] Step 2: 2-Bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)aniline AcOH (48.14 g, 801.67 mmol, 45.89 mL, 12.11 equivalents) was added to a solution of 4-(3-bromo-2-nitrophenoxy)tetrahydro-2H-pyran (20 g, 66.20 mmol, 1 equivalent) in ethyl acetate (180 mL) and H₂O (20 mL) at 25 °C. The mixture was allowed to be stirred at 50 °C. Fe (18.48 g, 330.99 mmol, 5 equivalents) was added at 50 °C. The mixture was allowed to be stirred at 80 °C for 2 h. The reactants were quenched by adding an aqueous solution of NaOH (10 mL) at 20 °C and the mixture was extracted with ethyl acetate (10 mL × 3). The organic solutions were combined, washed with brine (10 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residues were purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). 2-Bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)aniline was obtained as a yellow oil (18 g, 49.61 mmol, 74.94% yield, 75% purity). 1 H NMR (400 MHz, CDCl3) δ=7.06 (dd, J =0.8, 8.2 Hz, 1H), 6.76 (d, J =8.0Hz,1H), 6.60-6.53 (m, 1H), 6.39 (brs, 2H), 4.49 (tt, J =3.9, 8.0Hz, 1H), 4.02-3.96(m, 2H), 3.59 (ddd, J =3.2, 8.6, 11.7Hz, 2H), 2.08-2.04 (m, 2H), 1.83 (tdd, J =4.2, 8.6, 12.9Hz, 2H).

[0346] Step 3: N-(2-bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-4-chlorobutyramide 4-Chlorobutyryl chloride (5.60 g, 39.69 mmol, 4.44 mL, 1.2 equivalent) was added to a solution of 2-bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)aniline (9 g, 33.07 mmol, 1 equivalent) and Na2HPO4 (9.39 g, 66.14 mmol, 9.39 mL, 2 equivalent) in CHCl3 (90 mL), and the mixture was stirred at 25 °C for 2 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. N-(2-bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-4-chlorobutyramide (12 g, crude substance) was obtained as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ=7.24 (brd, J =9.5Hz, 1H), 7.12 (brt, J =6.4Hz, 1H), 6.90 (brd, J =8.1 Hz, 1H), 6.37 (brs, 2H), 4.51(dt, J =3.9, 7.6Hz, 1H), 3.99-3.90 (m, 2H), 3.60-3.54 (m, 2H), 2.16-2.06 (m,4H), 2.04-1.98 (m, 2H), 1.85-1.75 (m, 2H).

[0347] Step 4: 1-(2-bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)pyrrolidine-2-one N-(2-bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)-4-chlorobutyramide (10 g, 26.55 mmol, 1 equivalent) in DMF (50 mL) was added to a solution of NaH (1.17 g, 29.20 mmol, 60% purity, 1.1 equivalent) in DMF (50 mL) at 0 °C, and the mixture was allowed to be stirred at 25 °C for 12 h. The reaction mixture was quenched by adding saturated NH4Cl (50 mL) and extracted with ethyl acetate (50 mL × 2). The organic solutions were combined, washed with brine (30 mL), dried over Na2SO4, and concentrated. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). 1-(2-bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)pyrrolidine-2-one was obtained as a yellow oil (7 g, 16.46 mmol, 62.00% yield, 80% purity). 1H NMR (400 MHz, CDCl3) δ=7.25-7.19 (m, 1H),7.13 (t, J=8.2 Hz, 1H), 6.89 (d, J=8.2 Hz, 1H), 4.99-4.95 (m, 1H), 4.52 (tt,J=3.5, 7.0Hz, 1H), 4.35 (s, 1H), 3.96-3.87 (m, 2H), 3.59 (dddd, J=3.8, 7.5,11.4, 18.5Hz, 2H), 2.63-2.51 (m, 2H), 2.27-2.23 (m, 2H), 2.00 (td, J=3.4,6.9Hz, 2H), 1.82-1.75 (m, 2H).

[0348] Step 5: 1-(2-((tetrahydro-2H-pyran-4-yl)oxy)-6-vinylphenyl)pyrrolidine-2-one To a solution of 1-(2-bromo-6-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)pyrrolidine-2-one (3.5 g, 10.29 mmol, 1 equivalent) in dioxane (40 mL) and H₂O (4 mL), K₂CO₃ (4.27 g, 30.86 mmol, 3 equivalents), Pd(dppf)Cl₂ (1.51 g, 2.06 mmol, 0.2 equivalents), and trifluoro(vinyl)-λ₄-borane, potassium salt (4.13 g, 30.86 mmol, 3 equivalents) were added. The mixture was allowed to be stirred at 100 °C for 12 hours. The reactants were quenched at 20 °C by adding H₂O (30 mL) and extracted with ethyl acetate (15 mL × 3). The organic solutions were combined, washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). 1-(2-((tetrahydro-2H-pyran-4-yl)oxy)-6-vinylphenyl)pyrrolidine-2-one was given as a brown oil (47.36% yield). LCMS [M+1] = 288.1. 1 H NMR (400 MHz, CDCl3) δ=7.26-7.20 (m, 2H),6.87 (d, J =7.8 Hz, 1H), 6.71 (dd, J =11.1, 17.6Hz, 1H), 5.76 (d, J =17.5Hz, 1H), 5.34 (d, J =11.0Hz, 1H), 4.53 (tt, J =3.5, 7.0Hz, 1H), 3.96-3.89 (m, 2H), 3.83-3.76 (m, 1H), 3.65-3.50 (m, 3H), 2.61-2.54 (m, 2H), 2.24 (quin, J =7.5Hz, 2H), 2.05-1.95 (m, 2H), 1.84-1.76 (m, 2H).

[0349] Step 6: 2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)benzaldehyde At 0 °C, K₂O₄·2H₂O (179.51 mg, 487.21 μmol, 0.1 equivalent) and NaIO₄ (4.17 g, 19.49 mmol, 1.08 mL, 4 equivalent) were added to a solution of 1-(2-((tetrahydro-2H-pyran-4-yl)oxy)-6-vinylphenyl)pyrrolidine-2-one (1.4 g, 4.87 mmol, 1.13 mL, 2 equivalent) in THF (20 mL) and H₂O (10 mL). The mixture was stirred at 25 °C for 0.5 h. The reaction mixture was filtered and the filtrate was diluted with saturated Na₂SO₃ (20 mL) and extracted with ethyl acetate (10 mL × 3). The organic solutions were combined, washed with brine (15 mL × 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 1 / 0 to 0 / 1). 2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)benzaldehyde (950 mg, 2.63 mmol, 53.92% yield, 80% purity) was obtained as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ=10.03 (s, 1H), 7.54 (dd, J =1.1, 7.7Hz, 1H), 7.43 (t, J =7.9Hz, 1H), 7.26-7.20 (m, 1H), 4.61-4.55 (m, 1H), 4.03-3.91 (m, 3H), 3.67-3.57 (m, 3H), 2.66-2.53 (m, 2H), 2.34-2.25 (m, 2H), 2.12-2.04 (m, 2H), 1.83(brd, J =7.6Hz, 2H).

[0350] Step 7: 2-Hydroxy-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetonitrile TMSCN (488.62 mg, 4.93 mmol, 616.16 μL, 1.5 equivalents) and ZnCl2 (104.81 mg, 328.35 μmol, 0.1 equivalents) were added to a solution of 2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)benzaldehyde (950 mg, 3.28 mmol, 1 equivalent) in DCM (10 mL). The reaction mixture was allowed to be stirred at 25 °C for 12 hours. The reaction mixture was filtered, and the filtrate was diluted with H2O (10 mL) and extracted with ethyl acetate (10 mL × 3). The organic solutions were combined, washed with brine (10 mL × 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give 1.5 g of crude 2-hydroxy-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetonitrile as a yellow oil. LCMS [M+1] = 389.2.

[0351] Step 8: Methyl 2-hydroxy-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetate A solution of 2-hydroxy-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetonitrile (1.3 g, 2.01 mmol, 1 equivalent) in HCl / MeOH (6 M, 334.59 μL, 1 equivalent) was stirred at 30 °C for 24 h. The reaction mixture was concentrated under reduced pressure. The residue was analyzed by preparative HPLC (column: Phenomenex Luna C18 75). 30mm 3 μm; mobile phase: [H2O (0.1% TFA)-ACN]; gradient: 20%-40% B (purified for 8.0 min). 2-hydroxy-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetic acid methyl ester (57.03% yield) was obtained as a yellow oil. LCMS [M+1] = 350.1. 1 H NMR (400 MHz, CDCl3) δ=7.36 (t, J =8.1Hz, 1H), 7.08 (d, J =7.0Hz, 1H), 6.97 (d, J=8.2 Hz, 1H), 5.16 (s, 1H), 4.60-4.53(m, 1H), 3.95-3.69 (m, 9H), 2.72-2.61 (m, 2H), 2.33-2.22 (m, 2H), 2.13-1.98(m, 2H), 1.80 (ddd, J =3.5, 7.3, 13.6Hz, 2H).

[0352] Step 9: Methyl 2-((methylsulfonyl)oxy)-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetate TEA (130.33 mg, 1.29 mmol, 179.27 μL, 3 equivalents) and Ms₂O (224.37 mg, 1.29 mmol, 3 equivalents) were added to a solution of methyl 2-hydroxy-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetate (150 mg, 429.34 μmol, 1 equivalent) in DCM (3 mL) at 0 °C. The mixture was allowed to be stirred at 0 °C for 15 min. The reactants were quenched by adding NaHCO₃(aq) (2 mL) at 20 °C and extracted with DCM (2 mL × 3). The organic solutions were combined, washed with brine (3 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. Methyl 2-((methanesulfonyl)oxy)-2-(2-(2-oxopyrrolidone-1-yl)-3-((tetrahydro-2H-pyran-4-yl)oxy)phenyl)acetate (170 mg, crude) was obtained as a yellow oil. LCMS [M+1] = 428.1.

[0353] Preparation of tert-butyl 2-bromo-2-(5-(methoxymethyl)-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)acetate Step 1: 7-Bromo-2',3',5',6'-Tetrahydrospiro[indoline-3,4'-pyran]-2-one Three parallel reactions were carried out. A solution of 7-bromoindolin-2-one (15 g, 70.74 mmol, 1 equivalent) in DMF (450 mL) was added to a mixture of NaH (14.15 g, 353.70 mmol, 60% purity, 5 equivalents) in DMF (450 mL) under a N2 atmosphere at 0 °C. The reaction mixture was allowed to stir at 0 °C for 1 hour. 1-Bromo-2-(2-bromoethoxy)ethane (24.61 g, 106.11 mmol, 13.30 mL, 1.5 equivalents) was added at 0 °C. The reaction mixture was allowed to heat to 25 °C and stirred at 25 °C for 15 hours. The three parallel reactions were combined. The reaction mixture was quenched by adding H2O (3000 mL) and filtered. The filter cake was washed with water (600 mL × 3) and dried under vacuum to give 7-bromo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one as a red solid (50 g, 120.46 mmol, 56.76% yield, crude). LCMS [M+1] = 282.1.

[0354] Step 2: 7-Bromo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one Two parallel reactions were carried out. NaH (3.40 g, 85.07 mmol, 60% purity, 2 equivalents) was added to a solution of 7-bromo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (12 g, 42.53 mmol, 1 equivalent) in DMF (160 mL) at 0 °C under a N2 atmosphere. The reaction mixture was allowed to stir at 0 °C under a N2 atmosphere for 0.5 h. MeI (12.07 g, 85.07 mmol, 5.30 mL, 2 equivalents) was added dropwise to the reaction mixture at 0 °C. The reaction mixture was allowed to heat to 25 °C and stir for 2 h. The two reaction mixtures were combined and quenched by adding saturated NH4Cl (500 mL) at 0 °C. The mixture was extracted with ethyl acetate (50 mL × 3). The organic solutions were combined, washed with brine (50 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 3 / 1). 7-Bromo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one was given as a white solid (35.72% yield). LCMS [M+1] = 675.4. 1 H NMR (400 MHz, CDCl3) δ=7.40 (dd, J=0.8, 8.3Hz, 1H), 7.23 (dd, J =0.8, 7.4Hz, 1H), 6.93 (t, J =7.8 Hz, 1H), 4.32-4.26 (m,2H), 3.93-3.88 (m, 2H), 3.60 (s, 3H), 1.95-1.88 (m, 2H), 1.76 (brd, J=13.9Hz,2H).

[0355] Step 3: 7-Bromo-5-iodo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one Two parallel reactions were carried out. NIS (9.5 g, 42.21 mmol, 2.5 equivalents) was added to a solution of 7-bromo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (5 g, 16.88 mmol, 1 equivalent) in AcOH (50 mL) under a N2 atmosphere at 25 °C. The reaction mixture was heated to 50 °C and allowed to stir for 16 hours. The two reaction mixtures were combined, quenched by the addition of saturated NaHCO3 (200 mL), and extracted with ethyl acetate (30 mL × 3). The organic solutions were combined, washed with brine (30 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 0 to 90 / 10). 7-Bromo-5-iodo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one was given as a white solid (70.17% yield). LCMS [M+1] = 421.7. 1 HNMR (400 MHz, CDCl3) δ=7.73 (d, J =1.5Hz, 1H), 7.47 (d, J =1.5Hz, 1H), 4.29-4.23(m, 2H), 3.91-3.86 (m, 2H), 3.56 (s, 3H), 1.93-1.86 (m, 2H), 1.76-1.71 (m, 2H).

[0356] Step 4: 7-Bromo-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-5-carboxaldehyde Two parallel reactions were carried out. First, i-PrMgBr (1 M, 17.77 mL, 1.5 equivalents) was added dropwise to a solution of 7-bromo-5-iodo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (5 g, 11.85 mmol, 1 equivalent) in THF (50 mL) at 0 °C under a nitrogen atmosphere. The reaction mixture was allowed to stir at 0 °C for 0.5 h. Then, DMF (8.66 g, 118.47 mmol, 9.12 mL, 10 equivalents) was added to the reaction mixture at 0 °C. The reaction mixture was heated to 25 °C and allowed to stir at 25 °C for 2 h. The two reaction mixtures were combined, quenched at 20 °C by the addition of saturated NH4Cl (200 mL), and extracted with ethyl acetate (50 mL × 3). The organic solutions were combined, washed with brine (50 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 0 to 80 / 20). 7-Bromo-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-5-carboxaldehyde (78.12% yield) was given as a yellow solid. LCMS [M+1] = 323.8. 1 H NMR (400 MHz, CDCl3) δ=9.86 (s, 1H), 7.93 (d, J =1.3Hz, 1H), 7.75 (d, J =1.3Hz, 1H), 4.27 (t, J =9.8 Hz,2H), 3.95-3.91 (m, 2H), 3.65 (s, 3H), 1.99-1.94 (m, 2H), 1.77 (brd, J =13.9Hz, 2H).

[0357] Step 5: 7-Bromo-5-(hydroxymethyl)-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one Two parallel reactions were carried out. NaBH4 (350.12 mg, 9.25 mmol, 1 equivalent) was added to a solution of 7-bromo-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-5-carboxaldehyde (3 g, 9.25 mmol, 1 equivalent) in THF (30 mL) at 0 °C under a N2 atmosphere. The reaction mixture was heated to 25 °C and allowed to stir for 2 h. The two reaction mixtures were combined, quenched by adding water (250 mL) at 25 °C, and extracted with ethyl acetate (50 mL × 3). The organic solutions were combined, washed with brine (50 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 0 to 70 / 30). 7-Bromo-5-(hydroxymethyl)-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one was given as a yellow solid (82.82% yield). LCMS [M+1] = 325.9. 1 H NMR (400 MHz, CDCl3) δ=7.36 (s, 1H), 7.21 (s, 1H), 4.57 (s, 2H), 4.26-4.20 (m, 2H), 3.88-3.84 (m, 2H), 3.53 (s, 3H), 1.90-1.84 (m, 2H), 1.70(brd, J=13.9Hz, 2H).

[0358] Step 6: 7-Bromo-5-(methoxymethyl)-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one NaH (919.64 mg, 22.99 mmol, 60% purity, 1.5 equivalent) was added to a solution of 7-bromo-5-(hydroxymethyl)-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (5 g, 15.33 mmol, 1 equivalent) in THF (100 mL) at 0 °C under a N2 atmosphere. The reaction mixture was allowed to stir at 0 °C for 0.5 h. MeI (3.26 g, 22.99 mmol, 1.43 mL, 1.5 equivalent) was added to the reaction mixture at 0 °C. The reaction mixture was allowed to stir at 0 °C for 2 h. The reaction mixture was quenched at 25 °C by adding saturated NH4Cl (200 mL) and extracted with ethyl acetate (50 mL × 3). The organic solutions were combined, washed with brine (50 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 0 to 80 / 20). 7-Bromo-5-(methoxymethyl)-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one was given as a yellow solid (92.04% yield). LCMS [M+1] = 340.0.

[0359] Step 7: 2-(5-(methoxymethyl)-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)tert-butyl acetate Pd(t-Bu3P)3 (721.04 mg, 1.41 mmol, 0.1 equivalent) and ZnBrCH2COOt-Bu (1 M, 28.22 mL, 2 equivalent) were added to a solution of 7-bromo-5-(methoxymethyl)-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (4.8 g, 14.11 mmol, 1 equivalent) in THF (50 mL) under a N2 atmosphere. The reaction mixture was heated to 80 °C and stirred at 80 °C for 2 hours. The reaction mixture was quenched by adding water (150 mL) at 25 °C and filtered. The filter cake was washed with ethyl acetate (20 mL × 3) and then with 1 N HCl (500 mL). The filtrate was extracted with ethyl acetate (20 mL × 3). The organic solutions were combined, washed with brine (50 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 0 to 80 / 20). 2-(5-(methoxymethyl)-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)tert-butyl acetate was given as a yellow solid (84.95% yield). LCMS [M+1] = 376.3. 1 H NMR (400 MHz, CDCl3) δ=7.22 (s, 1H), 7.04 (s, 1H), 4.41 (s, 2H), 4.32-4.26 (m, 2H), 3.94-3.89 (m,2H), 3.80 (s, 2H), 3.44 (s, 3H), 3.40 (s, 3H), 1.96-1.90 (m, 2H), 1.75 (brd,J=13.9Hz, 2H), 1.45 (s, 9H).

[0360] Step 8: 2-Bromo-2-(5-(methoxymethyl)-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)tert-butyl acetate LDA (2 M, 1.60 mL, 1.2 equivalents) was added to a solution of 2-(5-(methoxymethyl)-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)tert-butyl acetate (1 g, 2.66 mmol, 1 equivalent) in THF (10 mL). The reaction mixture was allowed to stir at -70 °C for 0.5 h. TMSCl (434.04 mg, 4.00 mmol, 507.05 μL, 1.5 equivalents) was added to the reaction mixture at -70 °C. The reaction mixture was allowed to stir at -70 °C for 0.5 h. A solution of NBS (711.07 mg, 4.00 mmol, 1.5 equivalents) in THF (8 mL) was added to the reaction mixture at -70 °C. The reaction mixture was allowed to stir at -70 °C for 1 h. The reactants were quenched at 25 °C with saturated NH₄Cl (50 mL) and extracted with ethyl acetate (10 mL × 3). The organic solutions were combined, washed with brine (10 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 0 to 75 / 25). 2-Bromo-2-(5-(methoxymethyl)-1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)tert-butyl acetate (24.79% yield) was given as a yellow solid. LCMS [M+1] = 454.0. 1 H NMR (400 MHz, CDCl3) δ=7.53 (s, 1H), 7.28 (brs, 1H), 6.00 (s, 1H), 4.42 (s, 2H), 4.30-4.27 (m, 2H), 3.93-3.90 (m, 2H), 3.59 (s, 3H), 3.40 (s, 3H), 1.96-1.92 (m, 2H), 1.78-1.75 (m, 2H), 1.49 (s, 9H).

[0361] Preparation of methyl 2-(4-methylchroman-5-yl)acetate Step 1: Methyl (E)-3-(2-bromo-6-fluorophenyl)but-2-enoate t-BuOK (1 M, 82.94 mL, 1.2 equivalents) was added to a solution of methyl 2-(dimethoxyphosphoryl)acetate (15.10 g, 82.94 mmol, 11.95 mL, 1.2 equivalents) in THF (150 mL) at 0 °C. The mixture was allowed to stir at 0 °C for 0.5 h. 1-(2-bromo-6-fluorophenyl)ethyl-1-one (15 g, 69.11 mmol, 1 equivalent) was added to THF (150 mL) at 0 °C, and the mixture was allowed to stir at 0 °C for 1.5 h, then stirred at 50 °C for 14 h. The reactants were quenched by adding H2O (300 mL) and ethyl acetate (50 mL) was added. The organic solution was separated and the aqueous solution was extracted with ethyl acetate (50 mL × 2). The organic solutions were combined, washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was subjected to column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1, TLC: petroleum ether / ethyl acetate = 10 / 1, R) f = 0.56) Purification. Methyl (E)-3-(2-bromo-6-fluorophenyl)but-2-enoate (20 g, crude material) was obtained as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ=7.17-7.14 (m, 1H), 7.09-7.05 (m, 1H), 6.98-6.96(m, 1H), 6.01 (d, J =1.5Hz, 1H), 3.66 (s, 3H), 2.47 (d, J =1.1 Hz, 3H).

[0362] Step 2: (E)-3-(2-bromo-6-fluorophenyl)but-2-en-1-ol Two reactions were carried out in parallel. LiAlH4 (2.5 M, 24.17 mL, 1.5 equivalents) was added to a solution of methyl (E)-3-(2-bromo-6-fluorophenyl)but-2-enoate (11 g, 40.28 mmol, 1 equivalent) in THF (120 mL) under a nitrogen atmosphere at 0 °C. The reaction mixture was allowed to stir at 20 °C for 2 hours. H2O (2.5 mL) and NaOH (15%, 2.5 mL) were added to the reaction mixture. The mixture was further diluted with H2O (500 mL) and extracted with ethyl acetate (100 mL × 3). The organic solutions were combined, washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. (E)-3-(2-bromo-6-fluorophenyl)but-2-en-1-ol (50.65% yield) was given as a yellow oil.

[0363] Step 3: 5-bromo-4-methyl-2H-chromene Cs₂CO₃ (59.82 g, 183.61 mmol, 3 equivalents) was added to a solution of (E)-3-(2-bromo-6-fluorophenyl)but-2-en-1-ol (15 g, 61.20 mmol, 1 equivalent) in DMF (180 mL) at 20 °C. The mixture was allowed to stir at 80 °C for 16 h. The reactants were quenched by adding H₂O (400 mL) and ethyl acetate (200 mL) was added. The organic solution was separated and the aqueous solution was extracted with ethyl acetate (200 mL × 3). The organic solutions were combined, washed with brine (400 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). 5-Bromo-4-methyl-2H-chromene was given as a yellow oil (19.54% yield). 1 H NMR (400 MHz, CDCl3) δ=7.20 (dd, J =1.3, 7.9Hz, 1H), 6.97 (t, J =8.0Hz, 1H), 6.87 (dd, J =1.3, 8.1 Hz, 1H), 5.82 (ddd, J =1.5, 3.1, 6.0Hz, 1H), 4.48 (qd, J =1.4, 4.5Hz, 2H), 2.37 (q, J =1.5Hz, 3H).

[0364] Step 4: Methyl 2-(4-methyl-2H-chromene-5-yl)acetate A suspension of 5-bromo-4-methyl-2H-chromene (200 mg, 888.57 μmol, 1 equivalent), BrZnCH2COOMe (1 M, 4.44 mL, 5 equivalent), and Pd(t-Bu3P)2 (22.71 mg, 44.43 μmol, 0.05 equivalent) in THF (250 mL) was stirred at 70 °C under a nitrogen atmosphere for 2 hours. Water (5 mL) was added and the mixture was filtered. The filtrate was extracted with ethyl acetate (5 mL × 2). The organic solutions were combined, washed with brine (5 mL × 2), dried over Na2SO4, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, 25 g SepaFlash® silica gel rapid column, eluent 0–20% petroleum ether gradient / ethyl acetate, @ 120 mL / min). 2-(4-methyl-2H-chromen-5-yl)methyl acetate was obtained as a yellow oil (85.94% yield). LCMS [M+1] = 219.1. 1 H NMR (400 MHz, CDCl3) δ=7.10 (t, J=7.8 Hz, 1H), 6.87 (dd, J=1.1, 8.1 Hz, 1H), 6.80 (dd, J=0.8, 7.6Hz, 1H), 5.76 (ddd, J=1.4, 3.2,5.9Hz, 1H), 4.48-4.44 (m, 2H), 3.87-3.85 (m, 2H), 3.73-3.70 (m, 3H), 2.16 (d,J=1.4Hz, 3H).

[0365] Step 5: Methyl 2-(4-methylchroman-5-yl)acetate A solution of methyl 2-(4-methyl-2H-chromen-5-yl)acetate (2 g, 9.16 mmol, 1 equivalent) and Pd / C (20.00 g, 18.79 mmol, 1.88 mL, 10% purity, 2.05 equivalent) in MeOH (30 mL) was stirred at 30 °C under H2 (15 psi) for 12 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to give methyl 2-(4-methylchromen-5-yl)acetate (1.7 g, 84.22%) as a yellow oil. LCMS [M+1] = 221.2.

[0366] Preparation of 8-bromo-1-methylisochromo Step 1: 1-(2,6-dibromophenyl)ethyl-1-ol MeMgBr (3M, 142.09 mL, 1.5 equivalents) was added dropwise to a solution of 2,6-dibromobenzaldehyde (75 g, 284.18 mmol, 1 equivalent) in THF (1000 mL) at 0 °C. The mixture was allowed to stir at 0–10 °C for 2 hours. The reaction mixture was added dropwise to saturated NH4Cl (1000 mL) at 0 °C under a N2 atmosphere, and the mixture was extracted with ethyl acetate (200 mL × 2). The organic solutions were combined, washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give 1-(2,6-dibromophenyl)ethyl-1-ol (94.97% yield) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ=7.55 (d, J =8.0Hz, 2H), 6.97 (t, J =8.0Hz, 1H), 5.57 (q, J =6.8 Hz, 1H), 1.64 (d, J =6.8 Hz, 3H).

[0367] Step 2: 2-(1-(2,6-dibromophenyl)ethoxy)acetic acid NaH (10.29 g, 257.18 mmol, 60% purity, 2 equivalents) was added to a solution of 1-(2,6-dibromophenyl)ethyl-1-ol (36 g, 128.59 mmol, 1 equivalent) in THF (400 mL) at 0 °C under a N2 atmosphere. The mixture was allowed to stir at 0 °C for 0.5 h. BrCH2COOMe (39.34 g, 257.18 mmol, 24.35 mL, 2 equivalents) was added at 0 °C, and the solution was allowed to stir at 20 °C for 1 h. H2O (200 mL) and NaOH (10.29 g, 257.18 mmol, 2 equivalents) were added to the reaction mixture, and the mixture was allowed to stir at 20 °C for 2 h. The mixture was extracted with EtOAc (200 mL × 3). The aqueous solution was adjusted to pH 3 by adding HCl (1 M) and extracted with DCM (200 mL × 3). The organic solutions were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure to give 2-(1-(2,6-dibromophenyl)ethoxyacetic acid (98.93% yield) as a white solid.1 H NMR (400 MHz, CD3OD) δ=7.63 (d, J =8.0Hz, 2H), 7.09 (t, J =8.0Hz, 1H), 5.51-5.42 (m, 1H), 3.94 (d, J =16.5Hz, 1H), 3.70 (d, J =16.6Hz, 1H), 1.60 (d, J =6.8 Hz, 3H).

[0368] Step 3: 2-(1-(2,6-dibromophenyl)ethoxy)-N-methoxy-N-methylacetamide A mixture of 2-(1-(2,6-dibromophenyl)ethoxyacetic acid (96 g, 284.03 mmol, 1 equivalent) and HATU (118.80 g, 312.43 mmol, 1.1 equivalent) in DCM (1000 mL) was stirred at 20 °C for 1 h. DIEA (110.13 g, 852.09 mmol, 148.42 mL, 3 equivalent) and N,O-dimethylhydroxylamine hydrochloride (33.25 g, 340.84 mmol, 1.2 equivalent) were added to the mixture. The reaction mixture was allowed to stir at 20 °C for 15 h. The reaction mixture was quenched by adding H₂O (1000 mL) and extracted with ethyl acetate (200 mL × 3). The organic solutions were combined, washed with brine (200 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 3 / 1 to 1 / 1). 2-(1-(2,6-dibromophenyl)ethoxy)-N-methoxy-N-methylacetamide (85.93% yield) was obtained as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ=7.57 (d, J =8.1 Hz, 2H), 6.99 (t, J =7.9Hz, 1H), 5.45 (q, J =6.7Hz, 1H), 4.20 (brd, J =15.6Hz, 1H), 3.94 (d, J =15.8 Hz,1H), 3.59 (s, 3H), 3.19 (s, 3H), 1.68 (d, J =6.7Hz, 3H).

[0369] Step 4: 8-Bromo-1-methylisochroman-4-one n-BuLi (2.5 M, 53.53 mL, 1 equivalent) was added to a solution of 2-(1-(2,6-dibromophenyl)ethoxy)-N-methoxy-N-methylacetamide (51 g, 133.84 mmol, 1 equivalent) in THF (300 mL) at -70 °C, and the solution was allowed to be stirred at -70 °C under a N2 atmosphere for 2 h. The reaction mixture was quenched by adding saturated NH4Cl (300 mL) at 0 °C and extracted with ethyl acetate (50 mL × 3). The organic solutions were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1). 8-Bromo-1-methylisochroman-4-one (74.38% yield) was given as a white solid. 1 H NMR (400 MHz, CDCl3) δ=8.02 (dd, J =0.9, 7.8 Hz, 1H), 7.77 (dd, J =1.2, 7.9Hz, 1H), 7.30 (t, J =7.9Hz, 1H), 5.34-5.30 (m,1H), 4.64 (d, J =18.3Hz, 1H), 4.37 (d, J =18.1 Hz, 1H), 1.67 (d, J =6.8 Hz, 3H).

[0370] Step 5: 8-Bromo-1-methylisochromosome Et3SiH (92.85 g, 798.49 mmol, 127.54 mL, 5 equivalents) was added to a solution of 8-bromo-1-methylisochroman-4-one (38.5 g, 159.70 mmol, 1 equivalent) in TFA (300 mL) under a N2 atmosphere at 20 °C, and the solution was stirred at 50 °C for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was partitioned between ethyl acetate (20 mL) and saturated NaHCO3 (100 mL). The aqueous solution was extracted with ethyl acetate (20 mL × 2). The organic solutions were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give 8-bromo-1-methylisochroman-4-one as a white solid (82.72% yield). 1 H NMR (400 MHz, CDCl3)δ=7.39 (dd, J =0.6, 7.6Hz, 1H), 7.05 (td, J =7.2, 14.9Hz, 2H), 5.06 (q, J =6.5Hz, 1H), 4.11 (ddd, J =4.3, 9.4, 11.5Hz, 1H), 3.86 (ddd, J =3.8, 5.9, 11.5Hz, 1H),2.99-2.90 (m, 1H), 2.78-2.72 (m, 1H), 1.59 (d, J =6.5Hz, 3H).

[0371] Preparation of methyl 2-bromo-2-((S)-1,6-dimethylisochroman-8-yl)acetate Step 1: (S)-2-(1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)isocyano-8-yl)methyl acetate Pin2B2 (3.46 g, 13.62 mmol, 2 equivalents) and dtbpy (109.67 mg, 408.60 μmol, 0.06 equivalents) and [Ir(OMe)(cod)]2 (135.42 mg, 204.30 μmol, 0.03 equivalents) were added to a solution of (S)-2-(1,6-dimethylisochroman-8-yl)acetate (1.5 g, 6.81 mmol, 1 equivalent) in dioxane (20 mL) under a N2 atmosphere. The mixture was allowed to be stirred at 100 °C for 16 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give (S)-2-(1,6-dimethylisochroman-8-yl)acetate (3 g, crude matter) as a yellow oil. LCMS [M+1]=235.1. 1 H NMR (400 MHz, CDCl3)δ=7.51 (s, 2H), 5.15-5.08 (m, 1H), 4.15-4.06 (m, 1H), 3.83 (dd, J =5.0, 10.9Hz,1H), 3.71-3.69 (m, 3H), 3.65-3.61 (m, 1H), 3.60-3.55 (m, 1H), 2.93 (brdd, J =5.8, 7.8 Hz, 1H), 2.85-2.76 (m, 1H), 1.47 (d, J =6.5Hz, 3H), 1.34 (s, 12H).

[0372] Step 2: (S)-2-(1,6-dimethylisocyanate-8-yl)methyl acetate Pd(PPh3)4 (500.64 mg, 433.24 μmol, 0.05 equivalents), K2CO3 (3.59 g, 25.99 mmol, 3 equivalents), and MeI (6.15 g, 43.32 mmol, 2.70 mL, 5 equivalents) were added to a solution of (S)-2-(1,6-dimethylisocyanuric-8-yl)acetate (3 g, 8.66 mmol, 1 equivalent) in toluene (50 mL), EtOH (20 mL), and H2O (20 mL) under a N2 atmosphere at 20 °C. The reaction mixture was heated to 100 °C and stirred at 100 °C for 16 hours. The reaction mixture was quenched with water (200 mL) at 20 °C and extracted with ethyl acetate (100 mL × 3). The organic solutions were combined, washed with brine (100 mL × 3), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give (S)-2-(1,6-dimethylisochroman-8-yl)methyl acetate as a yellow oil (1 g, 3.41 mmol, 39.41% yield, 80% purity). LCMS [M+1] = 235.1. 1 H NMR (400 MHz, CDCl3) δ=6.93 (brs, 1H), 6.87 (s, 1H), 5.08 (q, J =6.5Hz, 1H), 4.13-4.05 (m,1H), 3.81 (td, J =5.1, 10.8 Hz, 1H), 3.71 (s, 3H), 3.63-3.57 (m, 1H), 3.53-3.48(m, 1H), 2.94-2.84 (m, 1H), 2.79-2.70 (m, 1H), 2.30 (s, 3H), 1.47 (brd, J =6.5Hz, 3H).

[0373] Step 3: Methyl 2-bromo-2-((S)-1,6-dimethylisocyano-8-yl)acetate LDA (2 M, 3.20 mL, 3 equivalents) was added to a solution of (S)-2-(1,6-dimethylisocyanuric-8-yl)acetate (0.5 g, 2.13 mmol, 1 equivalent) in THF (5 mL) at -60 °C, and the mixture was allowed to stir at -60 °C for 30 min. TMSCl (765.10 mg, 7.04 mmol, 893.81 μL, 3.3 equivalents) was added at -60 °C, and the mixture was allowed to stir at -60 °C for 0.5 h. A solution of NBS (569.74 mg, 3.20 mmol, 1.5 equivalents) in THF (5 mL) was added at -60 °C. The mixture was allowed to stir at 60 °C for 1 h. The reactants were diluted with saturated NH4Cl (50 mL) and extracted with ethyl acetate (20 mL × 3). The organic solutions were combined, washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 10 / 1 to 5 / 1) to give methyl 2-bromo-2-((S)-1,6-dimethylisochroman-8-yl)acetate as a yellow oil (0.46 g, 734.39 μmol, 34.41% yield, 50% purity).

[0374] Using a similar method, methyl 2-bromo-2-((R)-1,6-dimethylisochroman-8-yl)acetate can be prepared from any specified (R)-2-(1-methylisochroman-8-yl)acetate.

[0375] Preparation of tert-butyl 2-(1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)acetate Step 1: 7-Bromo-2',3',5',6'-Tetrahydrospiro[indoline-3,4'-pyran]-2-one Four parallel reactions were carried out. LiHMDS (1 M, 117.90 mL, 5 equivalents) was added to a solution of 7-bromoindolin-2-one (5 g, 23.58 mmol, 1 equivalent) in THF (100 mL) at -70 °C, and the mixture was allowed to stir at -70 °C for 0.5 h. Then, 1-bromo-2-(2-bromoethoxy)ethane (8.20 g, 35.37 mmol, 4.43 mL, 1.5 equivalents) was added at -70 °C. The reactants were allowed to stir at 25 °C for 16 h. The reactants were quenched by adding H₂O (500 mL) and ethyl acetate (200 mL) was added. The organic solutions were separated and the aqueous solution was extracted with ethyl acetate (200 mL × 3). The organic solutions were combined, washed with brine (500 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 3 / 1). 7-Bromo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one was given as a yellow solid (29.69% yield). 1 H NMR (400 MHz, CDCl3) δ=7.37 (d, J=8.3Hz, 1H), 7.29 (s, 1H), 6.97 (t, J=7.9Hz, 1H), 4.24 (td, J=6.2, 11.9Hz,2H), 3.92 (td, J=4.7, 11.8 Hz, 2H), 1.96-1.86 (m, 4H).

[0376] Step 2: 7-Bromo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one NaH (765.52 mg, 19.14 mmol, 60% purity, 1 equivalent) was added to a solution of 7-bromo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (5.4 g, 19.14 mmol, 1 equivalent) in DMF (55 mL) at 0 °C. The mixture was allowed to stir at 0 °C for 30 min and then MeI (5.43 g, 38.28 mmol, 2.38 mL, 2 equivalents) was added at 0 °C. The mixture was allowed to stir at 20 °C for 16 h. The mixture was quenched by adding saturated NH4Cl (200 mL) and then ethyl acetate (100 mL) was added. The organic and aqueous solutions were separated and the aqueous solution was extracted with ethyl acetate (100 mL × 3). The organic solutions were combined, washed with brine (200 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 10 / 1). 7-Bromo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one was given as a yellow solid (89.97% yield). 1 H NMR (400 MHz, CDCl3) δ=7.39(dd, J =1.1, 8.2 Hz, 1H), 7.22 (dd, J =1.0, 7.4Hz, 1H), 6.96-6.89 (m, 1H), 4.29(ddd, J =2.9, 10.4, 11.6Hz, 2H), 3.90 (td, J =4.3, 11.8 Hz, 2H), 3.59 (s, 3H), 1.97-1.88 (m, 2H), 1.80-1.72 (m, 2H).

[0377] Step 3: 2-(1-Methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)tert-butyl acetate BrZnCH2COOt-Bu (1 M, 43.90 mL, 2 equivalents) and Pd(t-Bu3P)2 (1.12 g, 2.19 mmol, 0.1 equivalents) were added to a solution of 7-bromo-1-methyl-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (6.5 g, 21.95 mmol, 1 equivalent) in THF (65 mL) at 20 °C. The mixture was allowed to be stirred at 80 °C for 2 h. The mixture was quenched by adding H2O and filtered. The filtrate was extracted with ethyl acetate (100 mL × 3). The organic solutions were combined, washed with brine (100 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1). 2-(1-methyl-2-oxo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-7-yl)tert-butyl acetate was obtained as a yellow solid (89.36% yield). 1 H NMR (400 MHz, CDCl3) δ=7.23 (dd, J =1.4, 7.1 Hz, 1H), 7.09-7.01 (m, 2H), 4.38-4.21 (m, 2H), 3.92 (td, J =4.3, 11.7Hz, 2H), 3.81 (s, 2H), 3.46 (s, 3H), 1.94 (ddd, J =4.4, 10.0, 14.1Hz, 2H), 1.77 (brd, J =13.9Hz, 2H), 1.45 (s, 9H).

[0378] Preparation of 4'-bromo-6'-methyl-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one Step 1: Methyl 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxoacetate LDA (2 M, 158.71 mL, 1.2 equivalents) was added dropwise to a solution of 1-bromo-3-fluoro-5-methylbenzene (50 g, 264.52 mmol, 1 equivalent) in THF (500 mL) at -70 °C under a N2 atmosphere. The reaction mixture was allowed to stir at -70 °C for 0.5 h. A solution of dimethyl oxalate (40.61 g, 343.87 mmol, 1.3 equivalents) in THF (500 mL) was added to the reaction mixture at -70 °C under a N2 atmosphere. The reaction mixture was allowed to stir at -70 °C under N2 for 2 h. The reactants were quenched by adding NH4Cl (500 mL) and extracted with EtOAc (200 mL × 3). The organic solutions were combined, washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 5 / 1). Methyl 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxoacetate was given as a yellow solid (90.71% yield). 1 H NMR (400 MHz, CDCl3) δ=7.28 (s, 1H), 6.95 (d, J =10.3Hz,1H), 3.94 (s, 3H), 2.40 (s, 3H).

[0379] Step 2: 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxo-N-(tetrahydro-2H-pyran-4-yl)acetamide Tetrahydro-2H-pyran-4-amine (12.13 g, 119.97 mmol, 1.1 equivalent) was added dropwise to a mixture of methyl 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxoacetate (30 g, 109.06 mmol, 1 equivalent) in MeOH (300 mL) at 0 °C. The mixture was allowed to be stirred at 25 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure. The crude product was milled with MeOH at 20 °C for 30 min. 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxo-N-(tetrahydro-2H-pyran-4-yl)acetamide (71.28% yield) was given as a yellow solid. 1 H NMR (400 MHz, CDCl3) δ=7.26-7.21 (m, 1H), 6.93 (d, J =9.5Hz, 1H), 6.85 (brd, J =6.7Hz, 1H), 4.13-3.97 (m,3H), 3.51 (dt,J =2.1, 11.7Hz, 2H), 2.38 (s, 3H), 2.04-1.94 (m, 2H), 1.70-1.55 (m, 2H).

[0380] Step 3: 4-Bromo-6-methyl-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione A solution of potassium tert-amyloxyacetamide (t-AmylOK) (4.56 g, 36.09 mmol, 5.24 mL, 1.3 equivalent) in toluene (50 mL) was added dropwise to a mixture of 2-(2-bromo-6-fluoro-4-methylphenyl)-2-oxo-N-(tetrahydro-2H-pyran-4-yl)acetamide (10 g, 27.76 mmol, 1 equivalent) in toluene (100 mL). The reaction mixture was allowed to be stirred at 0 °C for 2 h and then quenched by adding HCl (1 M, 200 mL). The mixture was extracted with EtOAc (100 mL × 3). The organic solutions were combined, dried over Na2SO4, filtered, and concentrated. The crude product was ground with petroleum ether:ethyl acetate (2 / 1, 50 mL). The mixture was filtered and the filtrate was concentrated to dryness to give 4-bromo-6-methyl-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione as a red solid (94.44% yield).

[0381] Step 4: 4-Bromo-6-methyl-1-(tetrahydro-2H-pyran-4-yl)indoline-2-one A solution of 24 g (74.04 mmol, 1 equivalent) of 4-bromo-6-methyl-1-(tetrahydro-2H-pyran-4-yl)indoline-2,3-dione in ethylene glycol (250 mL) was added to NH₂NH₂·H₂O (51.60 g, 1.03 mol, 50 mL, 13.92 equivalent) at 25 °C under a nitrogen atmosphere. The mixture was allowed to be stirred at 130 °C for 4 hours. The reaction mixture was concentrated under reduced pressure and quenched by adding H₂O (300 mL). The mixture was extracted with ethyl acetate (100 mL × 3). The organic solutions were combined, washed with brine (100 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was used directly without further purification. 4-bromo-6-methyl-1-(tetrahydro-2H-pyran-4-yl)indoline-2-one (25 g, crude substance) was obtained as a yellow solid. LCMS :[M+1]=312.0.

[0382] Step 5: 4'-Bromo-6'-methyl-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one NaH (14.63 g, 365.75 mmol, 60% purity, 1 equivalent) was added to a solution of 4-bromo-6-methyl-1-(tetrahydro-2H-pyran-4-yl)indoline-2-one (22.69 g, 73.15 mmol, 1 equivalent) in DMF (250 mL) under a N2 atmosphere at 0 °C. The solution was allowed to stir at 0 °C for 0.5 h, and then 1,2-dibromoethane (68.71 g, 365.75 mmol, 27.59 mL, 5 equivalent) was added to the solution at 0 °C and the solution was allowed to stir at 20 °C for 2 h. The reaction mixture was quenched by adding saturated NH4Cl (300 mL) and extracted with EtOAc (100 mL × 3). The organic solutions were combined, washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated to give the crude product. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 100 / 1 to 2 / 1). 4'-bromo-6'-methyl-1'-(tetrahydro-2H-pyran-4-yl)spiro[cyclopropane-1,3'-indoline]-2'-one was given as a yellow solid (60.99% yield).

[0383] Preparation of methyl 2-bromo-2-((1S,4S)-4,7-dimethyl-5',6'-dihydro-2'H,4'H-spiro[isochroman-1,3'-pyran]-5-yl)acetate Step 1: 2-(2-bromo-4-methylphenyl)-1-((3aR,6S,7aS)-8,8-dimethyl-2,2-dioxotetrahydro-3H-3a,6-methylbridged benzo[c]isothiazol-1(4H)-yl)ethan-1-one PivCl (84.22 g, 698.48 mmol, 85.94 mL, 4 equivalents) in toluene (20 mL) was added dropwise to a solution of 2-(2-bromo-4-methylphenyl)acetic acid (40 g, 174.62 mmol, 1 equivalent) in toluene (400 mL), (3aR,6S,7aS)-8,8-dimethylhexahydro-3H-3a,6-methylbridged benzo[c]isothiazolium 2,2-dioxide (37.60 g, 174.62 mmol, 1 equivalent) and TEA (35.34 g, 349.24 mmol, 48.61 mL, 2 equivalents) in toluene (20 mL). The mixture was allowed to be stirred at 110 °C for 16 hours. The reaction mixture was quenched by adding H2O (1 L) at 0 °C and extracted with ethyl acetate (300 mL × 3). The organic solutions were combined, washed with brine (300 mL × 3), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 100 / 1 to 3 / 1). 2-(2-bromo-4-methylphenyl)-1-((3aR,6S,7aS)-8,8-dimethyl-2,2-dioxotetrahydro-3H-3a,6-methylbridged benzo[c]isothiazo-1(4H)-yl)ethyl-1-one (150 g, crude) was obtained as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ=7.40 (s,1H), 7.15-7.12 (m, 1H), 7.10-7.06 (m, 1H), 4.29 (d, J =17.3Hz, 1H), 4.05 (d, J =17.3Hz, 1H), 3.92 (dd, J =4.9, 7.8 Hz, 1H), 3.52 (q, J =13.8 Hz, 2H), 2.34-2.29(m, 3H), 2.24-2.13 (m, 1H), 2.07-2.00 (m, 1H), 1.95-1.86 (m, 3H), 1.46-1.40(m, 1H), 1.38-1.31 (m, 1H), 1.25-1.21 (m, 3H), 0.99 (s, 3H).

[0384] Step 2: (S)-2-(2-bromo-4-methylphenyl)-1-((3aR,6S,7aS)-8,8-dimethyl-2,2-dioxotetrahydro-3H-3a,6-methylbridged benzo[c]isothiazolyl-1(4H)-yl)prop-1-one Four batches were carried out in parallel. LiHMDS (1M, 98.04 mL, 1.1 equivalents) was added to a solution of 2-(2-bromo-4-methylphenyl)-1-((3aR,6S,7aS)-8,8-dimethyl-2,2-dioxotetrahydro-3H-3a,6-methylbridged benzo[c]isothiazo-1(4H)-yl)ethyl-1-one (38 g, 89.13 mmol, 1 equivalent) in THF (400 mL) at -78 °C under a N2 atmosphere, and the mixture was stirred for 30 min. HMPA (79.8...

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in: It has a 6- to 12-membered aryl ring structure or a 3- to 19-membered heterocyclic ring structure. in Optional ground cover halogen, C 1-6 Alkyl, C 1-6 Alkoxy, 3-8 membered heterocyclic ring structure, aryl or 5-6 membered heteroaryl substitution, wherein the C 1-6 Alkyl, the C 1-6 alkoxy, the 3-8 membered heterocyclic ring structure, the aryl group, or the 5- to 6-membered heteroaryl group optionally coated with one or more halogens, C 1-4 Alkoxy or C 1-6 Alkyl substitution, the C 1-6 Alkyl groups may optionally be substituted with one or more halogens; a is 1, 2, 3, 4, 5, 6, 7, or 8; and R a C 1-6 alkyl.

2. The compound according to claim 1, wherein the compound is a compound of formula (IA) or a pharmaceutically acceptable salt thereof: 。 3. The compound according to any one of claims 1-2, wherein It is phenyl or 6-membered heteroaryl.

4. The compound according to any one of claims 1-3, wherein for ,in R1 is C 1-4 alkoxy group; and R2 is a C group optionally substituted with one or more halogens. 1-4 Alkyl; or When R1 and R2 together form fused 5- to 8-membered heterocyclic alkyl rings, fused 5- to 6-membered heteroaryl rings, or fused 7- to 15-membered spirocyclic heterocyclic alkyl ring systems... It is a 9- to 19-membered heterocyclic group, wherein the fused ring is optionally surrounded by halogen, C 1-4 Alkyl, C 3-6 Cycloalkyl or 4- to 8-membered heterocyclic alkyl ring substituted, wherein the C 1-4 Alkyl, the C 3-6 The cycloalkyl or the 4- to 8-membered heterocycloalkyl ring is optionally surrounded by one or more halogens or C. 1-4 Alkyl substitution, the C 1-4 The alkyl group is optionally substituted with one or more halogens; and R3 is a C that is optionally substituted with one or more halogens. 1-4 alkyl.

5. The compound according to claim 4, wherein... R1 is a methoxy group; R2 is fluorine; and R3 is a C that is optionally substituted with one or more fluorine molecules. 1-4 alkyl.

6. The compound according to any one of claims 1-3, wherein for in R4 and R 14 Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 alkyl; X1 is CR 5a R 5b or NR 5c R 5a H or C optionally substituted with one or more halogens 1-4 Alkyl, and R 5b C 1-4 Alkyl or C 3-6 cycloalkyl, wherein the C 1-4 alkyl or the C 3-6 The cycloalkyl group is optionally substituted with one or more halogens; or R 5a and R 5b Together they form spirocyclic 3 to 8-membered heterocyclic alkyl or spirocyclic C 3-6 Cycloalkyl, wherein the 3 to 8-membered heterocycloalkyl or the C 3-6 The cycloalkyl group is optionally substituted with the following: C14 groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 6-membered spirocyclic heterocyclic alkyl or C 3-6 Spirocyclic cycloalkyl, the 4- to 6-membered spirocyclic heterocyclic alkyl or C 3-6 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 Alkyl groups may optionally be substituted with one or more halogens; R 5c C 1-4 Alkyl or C 3-6 cycloalkyl, wherein the C 1-4 alkyl or the C 3-6 The cycloalkyl group may optionally be substituted with one or more halogens; X2 is CR 6a R 6b -CH2CR 6a R 6b C=O, O or NR 6c ; R 6a and R 6b Each independently represents H and C. 1-4 Alkyl, 5- to 6-heteroaryl, 6-aryl, C 3-7 Cycloalkyl or 4- to 7-membered heterocycloalkyl, wherein the C 1-4 alkyl, the 5- to 6-membered heteroaryl, the 6-membered aryl, the C 3-8 The cycloalkyl group or the 4- to 7-membered heterocycloalkyl group is optionally converted by one or more halogens or C. 1-4 Alkyl substitution; R 6c C that is optionally substituted with one or more halogens 1-4 alkyl X3 is a direct bond, CR 7a R 7b , O or NR 7c ; R 7a and R 7b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 alkyl; R 7c C that is optionally substituted with one or more halogens 1-4 alkyl; X4 is CR 8a R 8b or NR 8c R 8a For H and R 8b C is a halogen or optionally substituted with one or more halogens. 1-4 alkyl; R 8a and R 8b Together they form spirocyclic 3 to 8-membered heterocyclic alkyl or C 3-6 Cycloalkyl, wherein the 3 to 8-membered heterocycloalkyl or the C 3-6 The cycloalkyl group is optionally substituted with the following: C14 groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 8-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl, the 4- to 8-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group is optionally substituted with one or more halogens; and R 8c For H, C 1-4 Alkyl, 4- to 8-membered heterocyclic alkyl or C 3-6 cycloalkyl, wherein the C 1-4 Alkyl, the 4- to 8-membered heterocyclic alkyl, or the C 3-6 The cycloalkyl group may optionally be substituted with one or more halogens; The conditions are that X2 and X3 cannot both contain O or N heteroatoms, and X3 and X4 cannot both contain O or N heteroatoms.

7. The compound according to claim 6, wherein... R 14 For H; and X1 is CR 5a R 5b And R 5a Let H be the number of 'R', and R be the number of 'R'. 5b It is methyl or cyclopropyl.

8. The compound according to any one of claims 6-7, wherein R4 is H, fluorine, or methyl; X1 is CR 5a R 5b And R 5a Let H be the number of 'R', and R be the number of 'R'. 5b It is methyl; X2 is 0 and X3 is CR. 7a R 7b And R 7a and R 7b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 Alkyl; or X3 is 0 and X2 is CR. 6a R 7b And R 6a and R 6b Each is independently H; and X4 is CR 8a R 8b ,and R 8a and R 8b Each is independently H or C that is optionally substituted with one or more halogens. 1-4 alkyl, or R 8a and R 8b Together they form 4 to 7-membered heterocyclic alkyl rings or C 3-7 Cycloalkyl rings, wherein the 4- to 7-membered heterocyclic alkyl group or the C 3-7 The cycloalkyl group is optionally substituted with: halogen; C-shaped groups optionally substituted with one or more halogens. 1-4 Alkyl; 4- to 7-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl, the 4- to 7-membered spirocyclic heterocyclic alkyl or C 3-7 Spirocyclic cycloalkyl groups are optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

9. The compound of claim 6 or a pharmaceutically acceptable salt thereof, wherein... for in R 10a and R 10b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 Alkyl; or R 10a and R 10b Together they form a spiral ring C 3-6 cycloalkyl or spirocyclic 6-membered heterocyclic alkyl; R 11a and R 11b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 alkyl; R 12a and R 12b Each of the following is independently H, a halogen, or C that is optionally substituted with one or more halogens. 1-4 alkyl; R 13a It is methyl, ethyl, or cyclopropyl; R 13b For H; R 14 C is H, a halogen, or optionally substituted with one or more halogens. 1-4 alkyl.

10. The compound according to claim 9, wherein... R 10a and R 10b Each can be independently H or methyl; R 11a and R 11b Each can be independently H, methyl, or ethyl; R 12a and R 12b Each can be independently H or methyl; R 13a It is methyl, ethyl, or cyclopropyl; R 13b For H; R 14 It can be H, fluorine, or methyl.

11. The compound according to claim 10, wherein... R4 is H or fluorine; R 13a It is methyl, ethyl, or cyclopropyl; and R 14 For H.

12. The compound according to claim 6, wherein X2 is NR. 6c And R 6c C 1-4 alkyl.

13. The compound according to claim 6, wherein... for , where R 13a It is H or methyl, and R4 is H, F or methyl.

14. The compound according to claim 6, wherein... for R4 can be H, fluorine, or methyl.

15. The compound according to claim 14, wherein R 10a R 10b and R 11a Each is either H or methyl.

16. The compound according to claim 14, wherein R 11b It can be H, methyl, or ethyl.

17. The compound according to claim 6, wherein... for in R4 is H, a halogen, or a methyl group optionally substituted with one or more halogens; and R 9a and R 9b Each is independently H, a halogen, or a methyl group optionally substituted with one or more halogens.

18. The compound according to claim 17, wherein R4 is H, fluorine, or methyl; and R 9a and R 9b Each of the following is independently H, fluorine, chlorine, or a methyl group optionally substituted with one or more fluorine molecules.

19. The compound according to any one of claims 1-4, wherein for R 20 H, halogen, C 1-4 Alkyl or C 1-4 Alkoxy, wherein the C 1-4 alkyl groups and the C 1-4 Each alkoxy group may optionally be replaced by one or more halogens; X5 is a CR 25a R 25b or NR 25c R 25a and R 25b Each is independently H or optionally substituted with one or more halogens or alkoxy groups. 1-4 Alkyl; or R 25a and R 25b Together they form C 3-6 Cycloalkyl rings or 4- to 6-membered heterocyclic rings, each optionally coated with halogen, C 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or spirocyclic 4- to 6-membered heterocyclic ring substitution, wherein the C 1-4 Alkyl group, the spirocyclic C 3-6 The cycloalkyl or spirocyclic 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens or C. 1-4 Alkyl substitution; R 25c C 1-4 alkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 The alkyl group, the 5- to 6-membered heteroaryl group, or the C6 aryl group is optionally oxidized by one or more halogens or C6 alkyl groups. 1-4 Alkyl substitution; X6 is CR 26a R 26b Or C=O; R 26a and R 26b Each is independently H or optionally by one or more halogens or C 1-4 Alkyl-substituted C 1-4 alkyl; X7 is a CR 27a R 27b or NR 27c ; R 27a and R 27b Each is independently H or optionally substituted with one or more halogens or alkoxy groups. 1-4 alkyl; R 27a and R 27b Together they form C 3-6 cycloalkyl ring or 4- to 6-membered heterocyclic ring, wherein the C 3-6 The cycloalkyl ring or the 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens, one or more C... 1-4 Alkyl, spirocyclic C 3-6 Cycloalkyl or spirocyclic 4- to 6-membered heterocyclic ring substitution, wherein the C 1-4 Alkyl group, the spirocyclic C 3-6 The cycloalkyl or spirocyclic 4- to 6-membered heterocyclic ring is optionally surrounded by one or more halogens or C. 1-4 Alkyl substitution; and R 27c For H, C 1-4 Alkyl, C 3-6 Cycloalkyl, 4- to 7-membered heterocycloalkyl or 5- to 11-membered spirocycloheterocycloalkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 Alkyl, the C 3-6 Cycloalkyl, the 4- to 6-membered heterocycloalkyl, the 5- to 11-membered spirocycloheterocycloalkyl, the 5- to 6-membered heteroaryl, or the C6 aryl, optionally oxidized by one or more halogens or C 1-4 Alkyl substitution, the C 1-4 Alkyl groups may optionally be substituted with one or more halogens; The condition is that only one of X5 and X7 contains N heteroatoms.

20. The compound according to claim 19, wherein... R 20 H, halogen, C 1-4 Alkyl or C 1-4 Alkoxy, wherein the C 1-4 alkyl groups and the C 1-4 The alkoxy group may optionally be replaced by one or more halogens; X5 is a CR 25a R 25b ; R 25a and R 25b Each is independently a methyl group; or R 25a and R 25b Together they form a cyclopropyl group; X6 is C=O; R 26a and R 26b Each is independently H or optionally by one or more halogens or C 1-4 Alkyl-substituted C 1-4 alkyl; X7 is NR 27c ;and R 27c C 1-4 Alkyl, C 3-6 Cycloalkyl, 4- to 7-membered heterocycloalkyl or 5- to 11-membered spirocycloheterocycloalkyl, 5- to 6-membered heteroaryl or C6 aryl, wherein the C 1-4 Alkyl, the C 3-6 Cycloalkyl, the 4- to 6-membered heterocycloalkyl, the 5- to 11-membered spirocycloheterocycloalkyl, the 5- to 6-membered heteroaryl, or the C6 aryl, optionally oxidized by one or more halogens or C 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

21. The compound according to claim 20, wherein R 27c C 1-4 alkyl.

22. The compound according to claim 20, wherein R 27c It is a 5-membered heteroaryl group.

23. The compound according to claim 20, wherein R 27c To be optionally subjected to one or more halogens or C 1-4 Alkyl-substituted C 3-6 cycloalkyl, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

24. The compound according to claim 20, wherein R 27c C 1-4 Alkyl, 4- to 7-membered heterocyclic alkyl, or 5- to 11-membered spirocyclic heterocyclic alkyl, wherein the C 1-4 Alkyl groups, the 4- to 7-membered heterocyclic alkyl groups, and the 5- to 11-membered spirocyclic heterocyclic alkyl groups are optionally oxidized by one or more halogens or C. 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

25. The compound according to claim 24, wherein... for Where R 20 It can be H, fluorine, methyl, ethyl, methoxy, -CH2-O-CH3, or -CF3.

26. The compound according to claim 25, wherein R 27c It is a tetrahydropyran that is optionally substituted with one or more methyl groups or fluorine groups.

27. The compound according to claim 25, wherein R 27c for or ,in R 32a R 32b R 33a and R 33b Each independently is H, methyl, or fluorine; or R 32a and R 32b Or R 33a and R 33b One of them together forms a spiral ring C 3-6 Cycloalkyl or spirocyclic 3 to 6-membered heterocyclic rings.

28. The compound according to claim 25, wherein R 27c for .

29. The compound according to claim 19, wherein... for in R 30a R 30b R 31a and R 31b Each can be independently H, fluorine, or methyl; R 20 It can be H, fluorine, methyl, ethyl, methoxy, -CH2-O-CH3, or -CF3; and R 27c It is a methyl group.

30. The compound according to claim 25, wherein R 27c C 1-4 alkyl.

31. The compound according to claim 19, wherein... for .

32. The compound according to any one of claims 1-4, wherein for in R 20 For H; halogen; optionally C 1-4 alkoxy-substituted C 1-4 alkoxy group; C group optionally substituted with a 4- to 6-membered heterocyclic group or one or more halogens 1-4 alkyl.

33. The compound according to claim 32, wherein... R 25c C 1-4 Alkyl; and R 27c To be optionally subjected to one or more halogens or C 1-4 alkoxy-substituted C 1-4 Alkyl; C 3-6 Cycloalkyl or 4- to 6-membered heterocycloalkyl, wherein the C 3-6 Cycloalkyl or the 4- to 6-membered heterocycloalkyl group optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

34. The compound according to claim 32, wherein... R 27c C 1-4 Alkyl; and R 25c To be optionally subjected to one or more halogens or C 1-4 alkoxy-substituted C 1-4 Alkyl; C 3-6 Cycloalkyl or 4- to 6-membered heterocycloalkyl, wherein the C 3-6 Cycloalkyl or the 4- to 6-membered heterocycloalkyl group optionally C 1-4 Alkyl substitution, the C 1-4 The alkyl group may optionally be replaced by one or more halogens.

35. The compound according to any one of claims 32-34, wherein R 20 It is H or methyl.

36. The compound according to claim 19, wherein... for in R 20 For H; halogen; optionally C 1-4 alkoxy-substituted C 1-4 alkoxy group; C group optionally substituted with a 4- to 6-membered heterocyclic group or one or more halogens 1-4 alkyl; R 25c C 1-4 Alkyl; and R 27c It is a 4- to 6-membered heterocyclic group or C 3-6 cycloalkyl, wherein the 4- to 6-membered heterocyclic group or the C 3-6 cycloalkyl groups are optionally C 1-4 Alkyl substitution.

37. A chemical formula Compounds or their pharmaceutically acceptable salts.

38. A chemical formula Compounds or their pharmaceutically acceptable salts.

39. A chemical formula Compounds or their pharmaceutically acceptable salts.

40. A chemical formula Compounds or their pharmaceutically acceptable salts.

41. A chemical formula Compounds or their pharmaceutically acceptable salts.

42. A chemical formula Compounds or their pharmaceutically acceptable salts.

43. A chemical formula Compounds or their pharmaceutically acceptable salts.

44. A chemical formula Compounds or their pharmaceutically acceptable salts.

45. A chemical formula Compounds or their pharmaceutically acceptable salts.

46. ​​A chemical formula Compounds or their pharmaceutically acceptable salts.

47. A chemical formula Compounds or their pharmaceutically acceptable salts.

48. A chemical formula Compounds or their pharmaceutically acceptable salts.

49. A chemical formula Compounds or their pharmaceutically acceptable salts.

50. A chemical formula Compounds or their pharmaceutically acceptable salts.

51. A compound selected from the compounds in Figure 1 or pharmaceutically acceptable salts thereof.

52. A compound selected from the compounds in Table 3 or their pharmaceutically acceptable salts.

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