WO2024083086A1 - Dérivé bicyclique utilisé en tant qu'inhibiteur de l'intégrine - Google Patents

Dérivé bicyclique utilisé en tant qu'inhibiteur de l'intégrine Download PDF

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WO2024083086A1
WO2024083086A1 PCT/CN2023/124799 CN2023124799W WO2024083086A1 WO 2024083086 A1 WO2024083086 A1 WO 2024083086A1 CN 2023124799 W CN2023124799 W CN 2023124799W WO 2024083086 A1 WO2024083086 A1 WO 2024083086A1
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substituted
unsubstituted
ring
membered
heteroalkane
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Chinese (zh)
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郝宇
黄逸安
裴成奎
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上海如凌生物医药有限公司
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
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    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
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    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/438—The ring being spiro-condensed with carbocyclic or heterocyclic ring systems
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    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
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    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/538—1,4-Oxazines, e.g. morpholine ortho- or peri-condensed with carbocyclic ring systems
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    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
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    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/04—Ortho-condensed systems
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    • C07—ORGANIC CHEMISTRY
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    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07—Optical isomers

Definitions

  • the present invention relates to the field of medicinal chemistry, and in particular to a class of bicyclic derivatives as inhibitors of ⁇ v ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8 integrins, pharmaceutical compositions comprising such compounds, and their use in the treatment and/or prevention of conditions requiring inhibitors of ⁇ v ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8 integrins.
  • Integrins also known as integrins and junction proteins, are a large family of cell surface receptors. They are heterodimeric transmembrane glycoproteins formed by the combination of ⁇ subunits and ⁇ subunits. In mammals, 18 ⁇ and 8 ⁇ subunits have been found. These subunits are non-covalently bound to form at least 24 different integrins, which are differentially expressed by various cells and recognize a variety of ligands, mediating information transmission between cells and the extracellular matrix and between adjacent cells. Integrins are a type of cell adhesion molecules, consisting of three parts: the extracellular region, the transmembrane region, and the intracellular region.
  • the extracellular N-terminal domain binds to specific ligands, and the intracellular region is connected to the cytoskeleton protein through ⁇ -actinin, talin, and focal adhesion proteins to form a ligand-integrin-cytoskeleton transmembrane system.
  • integrin can transmit signals bidirectionally through the cell membrane. One is outside-in signal transmission. Integrins bind to ligands in the ECM, causing conformational changes in integrins, which aggregate on the cell membrane to form focal adhesions, activate multiple intracellular signaling pathways, and transduce signals from the extracellular to the intracellular, thereby regulating cell adhesion, diffusion, migration, proliferation, differentiation, and remodeling. The other is inside-out signaling.
  • Intracellular signals induce conformational changes in talin, causing talin to bind to the intracellular domain of the integrin ⁇ subunit. This binding unties the link between the ⁇ subunit tail and the ⁇ subunit tail in the cytoplasm, and the conformation of the integrin head changes, increasing its affinity for binding to extracellular ligands and enhancing the cell's adhesion ability.
  • integrins can be divided into three types: laminin-binding integrins, collagen-binding integrins, and arginine-glycine-aspartic acid sequence (Arg-Gly-Asp, RGD) integrins.
  • integrin ⁇ v ⁇ 1 is expressed on activated fibroblasts and mesangial cells; integrin ⁇ v ⁇ 6 can bind to the RGD sequence of TGF- ⁇ 1, achieve cell-cell contact and activate TGF- ⁇ 1; ⁇ v ⁇ 8 can bind to the RGD sequence in TGF- ⁇ 3LAP, present the LAP complex to matrix metalloproteinases on the cell surface, and activate TGF- ⁇ 1.
  • av ⁇ 6 and av ⁇ 1 play a major role in renal and pulmonary tissue fibrosis, while av ⁇ 1 plays a dominant role in liver fibrosis.
  • Integrin av ⁇ 1 is a low-affinity fibronectin receptor that is highly expressed in basal epithelial cells and has the function of promoting the migration of keratinocytes on the underlying fibronectin EDA. Blocking the interaction between integrin av ⁇ 1 and TGF- ⁇ 1 helps inhibit TGF- ⁇ 1 activity and block the process of fibrosis. It has been reported that the expression of integrin ⁇ v ⁇ 6 is very low in normal lung tissue, but when inflammation and fibrosis occur in lung injury, av ⁇ 6 is rapidly expressed (Hatley et al, Angewandte Chemie International Edition, 2018, 57(13): 3298.).
  • integrin av ⁇ 6 In patients with primary biliary cirrhosis (PBC), alcoholic fatty liver disease, hepatitis B, hepatitis C and other diseases, the mRNA expression of integrin av ⁇ 6 is increased. In chronic inflammatory and fibrotic diseases related to kidney disease, the expression of integrin av ⁇ 6 is significantly increased compared with normal kidney tissue. In addition, integrin av ⁇ 6 is significantly highly expressed in biopsy samples of patients with diabetes, Goodpasture's syndrome, Alport syndrome, lupus nephritis, etc. (Koivisto et al, The international journal of biochemistry & cell biology, 2018, 99: 186). Integrin ⁇ v ⁇ 8 is extremely important for the regulation of TGF ⁇ .
  • L-TGF ⁇ Its only binding ligand is L-TGF ⁇ , which is highly expressed in a variety of cancer cells, especially tumor cells with high Tregs. L-TGF ⁇ is mainly expressed in immune cells such as T cells.
  • ⁇ v ⁇ 8 promotes the differentiation of Tregs in the tumor microenvironment, thereby promoting tumor growth. The mechanism of action is that after T cell GARP/L-TGFbeta binds to ⁇ v ⁇ 8, Conformational changes occur, releasing activated TGF ⁇ , which binds to TGF ⁇ R on T cells and promotes the differentiation of T cells into Tregs.
  • conditional deletion of lung fibroblasts ⁇ v ⁇ 8 can inhibit mouse airway fibrosis.
  • Tissue fibrosis can occur in many organs and is a common fibrotic disease, including idiopathic pulmonary fibrosis (IPF), non-alcoholic fatty liver disease (NASH), cirrhosis, renal fibrosis, scleroderma, myocardial fibrosis, etc. Tissue damage and inflammation are important causes of fibrosis. Due to inflammation, organ parenchymal cells undergo necrosis, local immune cells are activated, and a variety of blood cells enter the site of injury. The activated immune cells produce a large number of highly biologically active cytokines and chemokines, leading to local activation of mesenchymal cells.
  • ECM extracellular matrix
  • cytokines cytokines
  • chemokines chemokines
  • angiogenic factors cytokines
  • angiogenic factors cytokines, chemokines, and angiogenic factors.
  • the abnormal increase and excessive deposition of extracellular matrix lead to pathological changes and tissue fibrosis (Ricard-Blumet al, Matrix Biology, 2018, 68: 122.).
  • the main feature of fibrosis is the formation and deposition of excessive fibrous connective tissue. Chronic fibrotic damage can destroy tissue structure, impair organ function, and eventually lead to organ failure.
  • the integrin family has attracted much attention as a key regulatory factor in chronic inflammation, fibrosis, and tumor immunity. There is an urgent need to develop a class of integrin inhibitors with novel structures that are safe and effective.
  • the purpose of the present invention is to provide a class of integrin inhibitors with novel structures of bicyclic derivatives.
  • Such compounds contain a bicyclic structure, have good inhibitory activity and certain drug selectivity against integrins ⁇ v ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8, excellent oral pharmacokinetic properties, and can effectively inhibit the increased expression of fibronectin and collagen in in vitro and in vivo pharmacodynamic models of fibrotic diseases.
  • Y1 is C1 ⁇ C6 alkylene, -O-, -(C1 ⁇ C6 alkylene)-O-, -NH-, -(C1 ⁇ C6 alkylene)-NH-;
  • Y2 is C1 ⁇ C6 alkylene, -O-, -(C1 ⁇ C6 alkylene)-O-, -C(O)-(C1 ⁇ C6 alkylene)-NH-, -NH-, -NH-(C1 ⁇ C6 alkylene)-, -(C1 ⁇ C6 alkylene)-NH-;
  • R1 is a substituted or unsubstituted 6-10 membered aromatic ring, a substituted or unsubstituted 5-8 membered heteroaromatic ring, a substituted or unsubstituted wherein the C ring and the D ring are each independently a substituted or unsubstituted C6-C10 aromatic ring, a substituted or unsubstituted 5-8 membered heteroaromatic ring, a substituted or unsubstituted 5-8 membered cycloalkane ring, or a substituted or unsubstituted 5-8 membered heteroalkane ring;
  • R2 is a hydrogen atom, a substituted or unsubstituted C6-C10 aromatic ring, a substituted or unsubstituted 5-8 membered heteroaromatic ring, a substituted or unsubstituted C8-C16 condensed ring, or -L1-L2;
  • -L1- is selected from none, -(substituted or unsubstituted C1-C6 alkylene)-, -(substituted or unsubstituted C1-C6 alkyleneoxy) -(substituted or unsubstituted C1-C6 alkylenethio)-, -(substituted or unsubstituted C3-C8 cycloalkyl)-, -(substituted or unsubstituted C3-C8 heterocycloalkyl)-, -(substituted or unsubstituted C6-C10 aryl)-, -(substituted or unsubstituted C5-C8 heteroaryl)-,
  • L2 is selected from none, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkylthio, substituted or unsubstituted C3-C8 cycloalkyl, -(C1-C3 alkyl)-substituted or unsubstituted C3-C8 cycloalkyl, -(C1-C3 alkoxy)-substituted or unsubstituted C3-C8 cycloalkyl, -O-substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heteroalkylcyclyl, -(C1-C3 alkyl)-substituted or unsubstituted C3-C8 heteroalkylcyclyl, -(C1-C3
  • X is an oxygen atom or a nitrogen atom
  • R 3a when X is an oxygen atom, R 3a is a hydrogen atom, a C1-C6 alkyl group, a substituted or unsubstituted C6-C10 aromatic ring, and R 3b is absent; when X is a nitrogen atom, R 3a is a hydrogen atom, a hydroxyl group, a C1-C6 alkyl group, a substituted or unsubstituted C6-C10 aromatic ring, and R 3b is a hydrogen atom;
  • A is a ring selected from the group consisting of a substituted or unsubstituted four-membered cycloalkane ring, a substituted or unsubstituted five-membered cycloalkane ring, a substituted or unsubstituted six-membered cycloalkane ring, a substituted or unsubstituted seven-membered cycloalkane ring, a substituted or unsubstituted eight-membered cycloalkane ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, a substituted or unsubstituted eight-membered heteroalkane ring,
  • B is a ring selected from the group consisting of a substituted or unsubstituted four-membered cycloalkane ring, a substituted or unsubstituted five-membered cycloalkane ring, a substituted or unsubstituted six-membered cycloalkane ring, a substituted or unsubstituted seven-membered cycloalkane ring, a substituted or unsubstituted eight-membered cycloalkane ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, a substituted or unsubstituted eight-membered heteroalkane ring,
  • A is a ring selected from the group consisting of a substituted or unsubstituted four-membered cycloalkane ring, a substituted or unsubstituted five-membered cycloalkane ring, a substituted or unsubstituted six-membered cycloalkane ring, a substituted or unsubstituted seven-membered cycloalkane ring, a substituted or unsubstituted eight-membered cycloalkane ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted a substituted seven-membered heteroalkane ring, a substituted or unsubstituted eight-membered heteroalkane
  • B is a ring selected from the group consisting of a substituted or unsubstituted four-membered cycloalkane ring, a substituted or unsubstituted five-membered cycloalkane ring, a substituted or unsubstituted six-membered cycloalkane ring, a substituted or unsubstituted seven-membered cycloalkane ring, a substituted or unsubstituted eight-membered cycloalkane ring, a substituted or unsubstituted C6-C10 aromatic ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, a substituted
  • n 0, 1, 2 or 3;
  • heteroaromatic ring, heteroalkyl ring or heteroaryl group each independently has 1-3 (preferably 1, 2 or 3) heteroatoms selected from N, O and S.
  • Y1 is none.
  • Y1 is C1 ⁇ C3 alkylene, -(C1 ⁇ C3 alkylene)-NH-.
  • Y 1 is C1-C3 alkylene.
  • Y1 is a methylene group.
  • Y2 is C1-C3 alkylene, -O-, -(C1-C3 alkylene)-O-, -C(O)-(C1-C3 alkylene)-NH-, -NH-, -NH-(C1-C3 alkylene)-, -(C1-C3 alkylene)-NH-.
  • Y2 is C1-C3 alkylene, -C(O)-(C1-C3 alkylene)-NH-, -(C1-C3 alkylene)-NH-.
  • Y2 is -O-, -(C1-C3 alkylene)-O-.
  • Y2 is C1-C3 alkylene.
  • Y2 is methylene, ethylene or -O-.
  • R 1 is substituted or unsubstituted
  • the C ring is a C6-C10 aromatic ring or a 5-8 membered heteroaromatic ring
  • the D ring is a substituted or unsubstituted 5-8 membered heteroaromatic ring or a substituted or unsubstituted 5-8 membered heteroalkyl ring.
  • R 1 is substituted or unsubstituted
  • the C ring is a C6 aromatic ring or a 5-6-membered nitrogen-containing heteroaromatic ring
  • the D ring is a substituted or unsubstituted 5-6-membered nitrogen-containing heteroaromatic ring or a substituted or unsubstituted 5-6-membered nitrogen-containing heteroalkane ring.
  • the nitrogen-containing heteroaromatic ring or nitrogen-containing heteroalkyl ring of the C ring or the D ring optionally further includes 1 or 2 O atoms.
  • R 1 is substituted or unsubstituted
  • the D ring is a substituted or unsubstituted 5-6 membered nitrogen-containing heteroaromatic ring or a substituted or unsubstituted 5-6 membered nitrogen-containing heteroalkane ring.
  • R 1 is a substituted or unsubstituted 5-8 membered nitrogen-containing heteroaromatic ring.
  • R 1 is a substituted or unsubstituted group selected from the following group:
  • R 1 is a substituted or unsubstituted pyridine ring, a substituted or unsubstituted 1,2,3,4-tetrahydronaphthyridine ring, a substituted or unsubstituted quinoline ring, or a substituted or unsubstituted azaindole ring.
  • R 1 is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • R 2 is a C6-C10 aromatic ring (preferably a C6 aromatic ring) substituted by R c , wherein R c is hydrogen, halogen, C1-C6 alkyl, C3-C8 cycloalkyl, or C1-C6 alkoxy.
  • the C8-C16 fused ring refers to a ring with 8 to 16 carbon atoms in which two or three rings are fused, and each ring in the fused ring is independently a 6-membered aromatic ring, a 5-6-membered heteroaromatic ring, a 5-6-membered cycloalkane ring, or a 5-6-membered heteroalkane ring.
  • R 2 is a C9-C10 fused ring.
  • the C9-C10 fused ring is preferably a 6-membered aromatic ring fused with a 5-6-membered heteroaromatic ring, more preferably a 6-membered aromatic ring fused with a 6-membered heteroaromatic ring.
  • R2 is a hydrogen atom or -L1-L2; wherein -L1- is -(substituted or unsubstituted C6-C10 aryl)-, and L2 is a substituted or unsubstituted 5-8 membered heteroaryl.
  • R2 is -L1-L2.
  • -L1- is selected from -(substituted or unsubstituted C3-C8 cycloalkyl)-, -(substituted or unsubstituted C3-C8 heterocycloalkyl)-, -(substituted or unsubstituted C6-C10 aryl)-, and -(substituted or unsubstituted C5-C8 heteroaryl)-.
  • -L1- is selected from -(substituted or unsubstituted C6-C10 aryl)-, -(substituted or unsubstituted 5-8 membered heteroaryl)-.
  • -L1- is selected from -(substituted or unsubstituted C6 aryl)-, -(substituted or unsubstituted 6-membered heteroaryl)-.
  • L2 is none, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkylthio, substituted or unsubstituted C3-C8 cycloalkyl, -(C1-C3 alkylene)-substituted or unsubstituted C3-C8 cycloalkyl, -(C1-C3 alkyleneoxy)-substituted or unsubstituted C3-C8 cycloalkyl, -O-substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted 3-8 membered heteroalkyl ring group, -(C1-C3 alkylene)-substituted or unsubstituted 3-8 membered heteroalkyl ring group, -(C1-C3 al
  • L2 is selected from substituted or unsubstituted 3-8 membered cycloalkyl, -(C1-C3 alkylene)-substituted or unsubstituted C3-C8 cycloalkyl, -(C1-C3 alkyleneoxy)-substituted or unsubstituted 3-8 membered cycloalkyl, -O-substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heteroalkyl ring group, -(C1-C3 alkylene)-substituted or unsubstituted 3-8 membered heteroalkyl ring group, -(C1-C3 alkyleneoxy)-substituted or unsubstituted 3-8 membered heteroalkyl ring group, -O-substituted or unsubstituted 3-8 membered hetero
  • L2 is selected from a substituted or unsubstituted 3-8 membered heteroalkyl ring group, and a substituted or unsubstituted 5-8 membered heteroaryl group.
  • -L1- is -(substituted or unsubstituted C6-C10 aryl)-
  • L2 is a substituted or unsubstituted 5-8 membered heteroaryl.
  • -L1- is -(substituted or unsubstituted phenyl)-
  • L2 is a substituted or unsubstituted 5-6-membered nitrogen-containing heteroaryl group (preferably pyrazolyl).
  • L1 is optionally substituted by one or more Ra , wherein Ra is halogen, C1-C6 alkyl, or C3-C6 cycloalkyl.
  • L1 is optionally substituted by one or more Ra , wherein Ra is halogen, C1-C6 alkyl, etc.
  • L2 is optionally substituted by one or more R b , wherein R b is C1-C3 alkyl.
  • R 2 is a structure of the following formula:
  • Z 1 , Z 2 , Z 3 , and Z 4 are each independently CR c or N;
  • L2 is none, C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkylthio, substituted or unsubstituted 3-8 membered cycloalkyl, substituted or unsubstituted 3-8 membered heteroalkyl ring group, substituted or unsubstituted 5-8 membered heteroaryl;
  • Each R c is independently hydrogen, halogen, C1-C6 alkyl, 3-8 membered cycloalkyl, or C1-C6 alkoxy.
  • R 2 is a structure of the following formula b:
  • Z 1 , Z 2 , Z 3 , Z 4 , Z 5 , Z 6 , Z 7 , Z 8 , and Z 9 are each independently CR c or N;
  • Each R c is independently hydrogen, halogen, C1-C6 alkyl, C3-C8 cycloalkyl, or C1-C6 alkoxy.
  • Z 1 , Z 2 , and Z 4 are each independently CR c , and Z 3 is CR c or N.
  • Z 1 , Z 2 , and Z 4 are each independently CH, and Z 3 is CR c or N.
  • Z 5 , Z 6 , and Z 7 are each independently CR c or N, and at least one of Z 5 , Z 6 , and Z 7 is N.
  • R 2 is the following structure:
  • L2 is C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkylthio;
  • Each R c is independently hydrogen, halogen, C1-C6 alkyl, C3-C8 cycloalkyl, or C1-C6 alkoxy.
  • each R c is independently fluorine, bromine, chlorine, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methoxy, or ethoxy.
  • R2 is substituted or unsubstituted
  • R2 is N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl-N-(2-aminoethyl)-2-aminoethyl
  • R 3a is a hydrogen atom or a C1-C3 alkyl group, and R 3b does not exist.
  • R 3a is a hydrogen atom
  • R 3b does not exist.
  • R 3a is a hydrogen atom, a hydroxyl group or a C1-C3 alkyl group, and R 3b is a hydrogen atom.
  • R 3a is a hydroxyl group
  • R 3b is a hydrogen atom
  • A is a ring selected from the group consisting of a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, and a substituted or unsubstituted eight-membered heteroalkane ring;
  • B is a ring selected from the group consisting of a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, and a substituted or unsubstituted eight-membered heteroalkane ring.
  • a and B are each independently a ring selected from the following group: a four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, and a substituted or unsubstituted six-membered heteroalkane ring.
  • the halogen is F, Cl, or Br, preferably F.
  • the heteroalkyl rings in the A ring and the B ring each independently contain 1-3 (preferably 1, 2, 3) nitrogen atoms.
  • the A ring and the B ring are each independently a ring selected from the following group: azetidine, substituted or unsubstituted pyrrolidine, substituted or unsubstituted piperidine ring, substituted or unsubstituted morpholine ring.
  • the substitution is preferably at the 3-position.
  • the substitution is preferably at the ortho position of the spiro atom.
  • the A ring and the B ring are each independently the following structure:
  • Y 1 is connected to the A ring through the nitrogen atom in the heteroalkyl ring.
  • the B ring is connected to Y 2 via a nitrogen atom in the heteroalkyl ring.
  • the compound is of the following formula II:
  • the number of ring atoms in the A ring is equal to or different from the number of ring atoms in the B ring.
  • the number of ring atoms in the A ring is less than, equal to or greater than the number of ring atoms in the B ring.
  • Y1 is connected to the A ring through the nitrogen atom in the heteroalkyl ring.
  • Y1 is -O-, -(C1-C6 alkylene)-O-, -NH- or -(C1-C6 alkylene)-NH-.
  • Y2 is connected to the B ring through the nitrogen atom in the heteroalkyl ring.
  • Y2 is -O-, -(C1-C6 alkylene)-O-, -NH- or -(C1-C6 alkylene)-NH-.
  • A is a 4-, 5-, 6-, 7- or 8-membered ring
  • B is a 4-, 5-, 6-, 7- or 8-membered ring.
  • the compound is of the following formula III-1:
  • the compound is of the following formula III-2:
  • the compound is of the following formula III-3:
  • the compound is of the following formula III-4:
  • the compound is of the following formula III-5:
  • the compound is of the following formula III-6:
  • R 1 , R 2 , Ring A and Ring B are as described above, and Y 1 and Y 2 are C1-C3 alkylene groups.
  • Y 1 , Y 2 , R 1 , R 2 , X, A, and B are each independently a corresponding group in Compound 1-122 prepared in Examples.
  • A is independently selected from the following rings: substituted or unsubstituted four-membered heteroalkane ring, substituted or unsubstituted five-membered heteroalkane ring, substituted or unsubstituted six-membered heteroalkane ring, substituted or unsubstituted seven-membered heteroalkane ring, substituted or unsubstituted five-membered heteroalkane ring and 6-10-membered aromatic ring, substituted or unsubstituted six-membered heteroalkane ring and 6-10-membered aromatic ring;
  • B is independently selected from the following rings: a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring and a 6-10-membered aromatic ring, and a substituted or unsubstituted six-membered heteroalkane ring and a 6-10-membered aromatic ring.
  • A is independently selected from the following rings: substituted or unsubstituted four-membered heteroalkane ring, substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring;
  • B is a ring independently selected from the group consisting of a substituted or unsubstituted C6-C10 aromatic ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, and a substituted or unsubstituted six-membered heteroalkane ring.
  • the "substituted" means that 1 to 4 (preferably 1, 2, 3 or 4) hydrogen atoms on the group are independently replaced by substituents selected from the following group: halogen, -OH, -CN, -NO 2 , -NH 2 , -NCO, -OCN, -SCN, -NCS, -N 3 , -(C1-C3 alkylene)OH, -O(C1-C3 alkyl), -S(C1-C3 alkyl), -C(O)X'R 4 or -X'C(O)R 5 , -SO 3 R 4 , -OSO 2 OR 4 , -NR 6 SO 2 OR 5 , -NR 6 R 7 , -SO 2 NR 6 R 7 , -NH-SO 2 -R 6 , -N + R 6 R 7 R 8 , -C(O)N(R 9 ) 2 , -SO 2 R 10 , -NR 6 SO 2 OR
  • X' is a chemical bond or is oxygen or sulfur
  • R4 is independently selected from hydrogen, C1-C6 alkyl, 3-12-membered alkyl ring group, 5-10-membered aryl group, 3-12-membered heterocyclic group
  • R5 is independently selected from hydrogen, C1-C6 alkyl, 3-12-membered alkyl ring group, 5-10-membered aryl group, 3-12-membered heterocyclic group;
  • R 6 , R 7 and R 8 are each independently selected from hydrogen, C1-C6 alkyl, -C(O)X'R 4 or -X'C(O)R 5 , or, R 6 and R 7 together with the nitrogen atom to which they are attached form a 4-8 membered heterocyclic ring in the ring structure; wherein, R 6 or R 7 are not all -C(O)X'R 4 or -X'C(O)R 5 ;
  • R 9 is independently selected from hydrogen or C1-C6 alkyl, or two R 9 together with the N atom to which they are attached form a 4-8 membered heterocyclic ring;
  • R 10 is independently selected from hydrogen, C1-C6 alkyl, 3-12 membered alkyl ring group, 3-12 membered heterocyclic group, 5-10 membered aryl group;
  • R 11 is independently selected from hydrogen, C1-C6 alkyl, or two R 11 together with the N atom to which they are attached form a 4-8 membered heterocyclic ring;
  • p is an integer of 0-3 for each independently.
  • the bicyclic derivative is selected from the following group:
  • the bicyclic derivative as described in the first aspect of the present invention there is provided a method for preparing the bicyclic derivative as described in the first aspect of the present invention, and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof,
  • the method comprises one of the following methods (1) to (5):
  • a pharmaceutical composition comprising:
  • a bicyclic derivative of formula (I) as described in the first aspect of the present invention and its racemate, stereoisomer, tautomer, isotope-labeled substance, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt thereof, for use in the preparation of a pharmaceutical composition for treating or preventing a disease, disorder or condition associated with the activity or expression of ⁇ v ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8 integrins.
  • the disease, disorder or condition associated with the activity or expression of ⁇ v ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8 integrins is selected from the group consisting of autoimmune diseases, fibrotic diseases, inflammatory bowel disease (IBD), relapsing multiple sclerosis (RMS), progressive multifocal leukoencephalopathy (PML), ulcerative colitis (UC), Crohn's disease (CD), chronic viral hepatitis B and C, non-alcoholic fatty liver disease (NAFLD), age-related macular degeneration, diabetic retinopathy, retinal vascular disease, osteoporosis and cell proliferative diseases.
  • IBD inflammatory bowel disease
  • RMS relapsing multiple sclerosis
  • PML progressive multifocal leukoencephalopathy
  • UC ulcerative colitis
  • CD Crohn's disease
  • NAFLD non-alcoholic fatty liver disease
  • age-related macular degeneration diabetic retinopathy
  • retinal vascular disease retinal vascular disease
  • the fibrotic disease is selected from the following group: pulmonary fibrosis, idiopathic pulmonary fibrosis, nonspecific interstitial pneumonia (NSIP), conventional interstitial lung disease (UIP), radiation-induced pulmonary fibrosis, familial pulmonary fibrosis, airway pulmonary fibrosis, chronic group A lung disease (COPD), interstitial lung disease, liver fibrosis, chronic kidney disease, renal fibrosis, skin fibrosis, systemic sclerosis, or a combination thereof.
  • pulmonary fibrosis idiopathic pulmonary fibrosis
  • NIP nonspecific interstitial pneumonia
  • UIP conventional interstitial lung disease
  • COPD chronic group A lung disease
  • interstitial lung disease liver fibrosis, chronic kidney disease, renal fibrosis, skin fibrosis, systemic sclerosis, or a combination thereof.
  • the fibrotic disease is pulmonary fibrosis (such as IPF), liver fibrosis, skin fibrosis, scleroderma, cardiac fibrosis, renal fibrosis, intestinal fibrosis, primary sclerosing cholangitis (PSC) or biliary fibrosis (such as PBC).
  • the fibrotic disease is pulmonary fibrosis (IPF).
  • IPF pulmonary fibrosis
  • the fibrotic disease is primary sclerosing cholangitis or biliary fibrosis (such as PBC).
  • the fibrotic disease is scleroderma.
  • the fibrotic disease is psoriasis.
  • the cell proliferative disease is cancer, selected from the following group: breast cancer, cervical cancer, endometrial cancer, ovarian cancer, colon cancer, rectal cancer, pancreatic cancer, liver cancer, lung cancer, non-small cell lung cancer, brain metastasis of lung cancer, oral squamous cell carcinoma, head cancer, neck cancer, head and neck squamous cell carcinoma, oral or nasal mucosal cancer, laryngeal cancer, kidney cancer, renal cell carcinoma, ovarian cancer, spleen cancer, small intestine cancer, large intestine cancer, gastric cancer, esophageal cancer, esophageal cancer, lung squamous cell carcinoma, bile duct cancer, gallbladder cancer, endometrial cancer, melanoma, urothelial carcinoma, urogenital tract cancer, genital cancer, prostate cancer, testicular cancer, bladder cancer, blood cancer, skin cancer, bone marrow cancer, brain cancer, central nervous system cancer, muscle tissue cancer, thyroid cancer,
  • a method for treating a disease, disorder or condition associated with the activity or expression of av ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8 integrins comprising the steps of administering an effective amount of a bicyclic derivative of formula (I) as described in the first aspect of the present invention, and its racemate, stereoisomer, tautomer, isotope label, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or a pharmaceutically acceptable salt thereof to a patient in need thereof.
  • a bicyclic derivative is provided, and its racemate, stereoisomer, tautomer, isotope label, nitrogen oxide, solvate, polymorph, metabolite, ester, prodrug or pharmaceutically acceptable salt, wherein the bicyclic derivative has the structure shown in the following formula II-a, II-b, II-c:
  • Each Y 1 is independently C1-C6 alkylene, -O-, -(C1-C6 alkylene)-O-, -NH-, -(C1-C6 alkylene)-NH-;
  • Each Y2 is independently C1-C6 alkylene, -O-, -(C1-C6 alkylene)-O-, -C(O)-(C1-C6 alkylene)- NH-, -NH-, -NH-(C1-C6 alkylene)-, -(C1-C6 alkylene)-NH-;
  • Each R 1 is independently a substituted or unsubstituted 6-10 membered aromatic ring, a substituted or unsubstituted 5-8 membered heteroaromatic ring, a substituted or unsubstituted wherein the C ring and the D ring are each independently a substituted or unsubstituted C6-C10 aromatic ring, a substituted or unsubstituted 5-8 membered heteroaromatic ring, a substituted or unsubstituted 5-8 membered cycloalkane ring, or a substituted or unsubstituted 5-8 membered heteroalkane ring;
  • Each R 2 is independently a hydrogen atom, a substituted or unsubstituted C6-C10 aromatic ring, a substituted or unsubstituted 5-8 membered heteroaromatic ring, a substituted or unsubstituted C8-C16 condensed ring, or -L1-L2;
  • -L1- is selected from none, -(substituted or unsubstituted C1-C6 alkylene)-, -(substituted or unsubstituted C1-C6 alkyleneoxy)-, -(substituted or unsubstituted C1-C6 alkylenethio)-, -(substituted or unsubstituted C3-C8 cycloalkyl)-, -(substituted or unsubstituted C3-C8 heterocycloalkyl)-, -(substituted or unsubstituted C6-C10 aryl)-, -(substituted or unsubstituted C5-C8 heteroaryl)-,
  • L2 is selected from none, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 alkylthio, substituted or unsubstituted C3-C8 cycloalkyl, -(C1-C3 alkyl)-substituted or unsubstituted C3-C8 cycloalkyl, -(C1-C3 alkoxy)-substituted or unsubstituted C3-C8 cycloalkyl, -O-substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heteroalkylcyclyl, -(C1-C3 alkyl)-substituted or unsubstituted C3-C8 heteroalkylcyclyl, -(C1-C3
  • Each R 3a ' is independently a C1-C6 alkyl group
  • each Each is independently a substituted or unsubstituted spiro ring or a substituted or unsubstituted cyclocyclic ring;
  • A is a ring selected from the group consisting of a substituted or unsubstituted four-membered cycloalkane ring, a substituted or unsubstituted five-membered cycloalkane ring, a substituted or unsubstituted six-membered cycloalkane ring, a substituted or unsubstituted seven-membered cycloalkane ring, a substituted or unsubstituted eight-membered cycloalkane ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, a substituted or unsubstituted eight-membered heteroalkane ring,
  • B is a ring selected from the group consisting of a substituted or unsubstituted four-membered cycloalkane ring, a substituted or unsubstituted five-membered cycloalkane ring, a substituted or unsubstituted six-membered cycloalkane ring, a substituted or unsubstituted seven-membered cycloalkane ring, a substituted or unsubstituted eight-membered cycloalkane ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, a substituted or unsubstituted eight-membered heteroalkane ring,
  • A is a ring selected from the group consisting of a substituted or unsubstituted four-membered cycloalkane ring, a substituted or unsubstituted five-membered cycloalkane ring, a substituted or unsubstituted six-membered cycloalkane ring, a substituted or unsubstituted seven-membered cycloalkane ring, a substituted or unsubstituted eight-membered cycloalkane ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, a substituted or unsubstituted eight-membered heteroalkane ring,
  • B is a ring selected from the group consisting of a substituted or unsubstituted four-membered cycloalkane ring, a substituted or unsubstituted five-membered cycloalkane ring, a substituted or unsubstituted six-membered cycloalkane ring, a substituted or unsubstituted seven-membered cycloalkane ring, a substituted or unsubstituted eight-membered cycloalkane ring, a substituted or unsubstituted C6-C10 aromatic ring, a substituted or unsubstituted four-membered heteroalkane ring, a substituted or unsubstituted five-membered heteroalkane ring, a substituted or unsubstituted six-membered heteroalkane ring, a substituted or unsubstituted seven-membered heteroalkane ring, a substituted
  • n is independently 0, 1, 2 or 3;
  • heteroaromatic ring, heteroalkyl ring or heteroaryl group each independently has 1 to 3 (preferably 1, 2 or 3) heteroatoms selected from N, O and S;
  • Formula II-a, Formula II-b and Formula II-c are all chemically stable structures.
  • Y 1 , Y 2 , R 1 , R 2 , A, B and n are independently as described in the first aspect of the present invention.
  • the "substituted" means that 1 to 4 (preferably 1, 2, 3 or 4) hydrogen atoms on the group are independently replaced by a Boc substituent.
  • the bicyclic derivative is selected from the following group:
  • FIG. 1 shows the results of Western blotting (WB) experiments of the compounds, illustrating the degradation effects of Examples 1, 2, 7, 15, and 25 on fibrosis proteins ⁇ -SMA and Fibronectin at different concentrations. Pirfenidone and GSK-3008348 were selected as control compounds in this experiment.
  • Figure 2 shows the body weight curves of rats in the sham operation group, model group, 100 mg/kg nintedanib group, 50 mg/kg low-dose group of Example 71 and 250 mg/kg high-dose group of Example 71 (mean ⁇ SEM).
  • Figure 3 shows the body weight growth rate curves (mean ⁇ SEM) of rats in the sham operation group, model group, 100 mg/kg nintedanib group, 50 mg/kg low-dose group of Example 71 and 250 mg/kg high-dose group of Example 71.
  • Figure 4 shows the histological changes of bronchioles and pulmonary arterioles in the left lung pulmonary fibrosis lesions in the sham operation group (A), model group (B), 100 mg/kg nintedanib group (C), 50 mg/kg Example 71 low-dose group (D), and 250 mg/kg Example 71 high-dose group (E) ( ⁇ 200, H&E staining);
  • Figure A shows normal pulmonary bronchioles and accompanying pulmonary arterioles;
  • Figure B shows damage to bronchioles and pulmonary arterioles;
  • Figures C-E show that the degree of damage to bronchioles and pulmonary arterioles is reduced.
  • Figure a is a pulmonary arteriole;
  • b is a bronchiole.
  • the arrows in Figures B-E indicate inflammatory cell infiltration.
  • Figure 5 shows the injury and inflammation scores of the terminal bronchioles and the accompanying small pulmonary arteries in the left lung pulmonary fibrosis lesions in the sham operation group, the model group, the 100 mg/kg nintedanib group, the 50 mg/kg low-dose group of Example 71, and the 250 mg/kg high-dose group of Example 71.
  • *** is p ⁇ 0.001 (compared with the model group).
  • Figure 6 shows the histological changes of bronchioles and pulmonary arterioles at the edge of the left lung pulmonary fibrosis lesions in the sham operation group (A), model group (B), 100 mg/kg nintedanib group (C), 50 mg/kg Example 71 low-dose group (D), and 250 mg/kg Example 71 high-dose group (E) ( ⁇ 200, H&E staining).
  • Figure A normal pulmonary bronchioles and accompanying pulmonary arterioles can be seen
  • Figure B shows damage to bronchioles and pulmonary arterioles
  • Figures C-E show that the degree of damage to bronchioles and pulmonary arterioles is reduced.
  • a is a pulmonary arteriole
  • b is a bronchiole.
  • the arrows in Figures B-E indicate inflammatory cell infiltration.
  • Figure 7 shows the injury and inflammation scores of the terminal bronchioles and the accompanying small pulmonary arteries at the edge of the left lung pulmonary fibrosis lesions in the sham operation group, the model group, the 100 mg/kg nintedanib group, the 50 mg/kg low-dose group of Example 71, and the 250 mg/kg high-dose group of Example 71.
  • *** is p ⁇ 0.001
  • ** is p ⁇ 0.01 (compared with the model group).
  • Figure 8 shows the histological changes of alveolar tissue in the left lung pulmonary fibrosis lesions in the sham operation group (A), model group (B), 100 mg/kg nintedanib group (C), 50 mg/kg low-dose group of Example 71 (D), and 250 mg/kg high-dose group of Example 71 (E) ( ⁇ 200, H&E staining).
  • Figure A normal alveolar walls can be seen;
  • Figure B shows that the sheet-like alveolar structure in the fibrotic lesions is damaged and disappeared, filled with a large number of exuded inflammatory cells and proliferating connective tissue.
  • the alveolar structure in Figures C-E is abnormal, it is significantly better than that in the model group.
  • Group E that is, the high-dose group of Example 71, some alveoli are close to the normal alveolar wall structure.
  • the arrows in Figures B-E indicate that the alveolar structure in the fibrotic lesions is damaged and the remaining alveolar wall is thickened.
  • Figure 9 shows alveolar fibrosis lesions in the left lung pulmonary fibrosis lesions ( ⁇ 200, Masson staining) in the sham operation group (A), model group (B), 100 mg/kg nintedanib group (C), 50 mg/kg Example 71 low-dose group (D), and 250 mg/kg Example 71 high-dose group (E).
  • Normal physiological fibrous tissue without fibrous tissue deposition can be seen in Figure A; severe fibrotic lesions and obvious fibrous tissue deposition can be seen in alveolar tissue in Figure B.
  • Figures C-E also have fibrotic lesions, but are significantly better than the model group.
  • the arrows in Figures B-E indicate fibrous tissue deposition.
  • Figure 10 shows the left lung pulmonary fibrosis scores of the sham group, model group, 100 mg/kg nintedanib group, 50 mg/kg low-dose group of Example 71, and 250 mg/kg high-dose group of Example 71.
  • *** means p ⁇ 0.001
  • * means p ⁇ 0.05 (compared with the model group).
  • Figure 11 shows the proportion of different pathological scores of left lung pulmonary fibrosis in the sham operation group, model group, 100 mg/kg nintedanib group, 50 mg/kg low-dose group of Example 71, and 250 mg/kg high-dose group of Example 71.
  • *** indicates p ⁇ 0.001 (compared with the model group).
  • Figure 12 shows the degree of collagen deposition in the left lung fibrosis lesions in the sham operation group (A), model group (B), 100 mg/kg nintedanib group (C), 50 mg/kg low-dose group of Example 71 (D), and 250 mg/kg high-dose group of Example 71 (E) ( ⁇ 200, Masson staining).
  • Figure A normal physiological fibrous tissue exists, and the collagen area is ⁇ 5%;
  • Figure B shows extremely serious collagen deposition.
  • Figures C-E also show an increase in collagen deposition, but the degree is lower than that of the model group.
  • Group E that is, the high-dose group of Example 71, has the lightest collagen deposition, which is significantly lower than the model group and also lower than the nintedanib group.
  • Figure 13 shows the percentage of collagen deposition area in the sham operation group, model group, 100 mg/kg nintedanib group, 50 mg/kg low-dose group of Example 71, and 250 mg/kg high-dose group of Example 71.
  • *** means p ⁇ 0.001
  • * means p ⁇ 0.05 (compared with the model group).
  • the bicyclic derivatives have excellent activity and selectivity for integrins ⁇ v ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8, and oral pharmacokinetic properties that are significantly better than the positive control PLN-74809, and show activity effects that are better than the positive references pirfenidone and nintedanib in in vitro and in vivo pharmacodynamic models of fibrotic diseases, so they can be used to prepare and prevent pharmaceutical compositions for fibrotic diseases. Based on the above findings, the inventors completed the present invention.
  • the terms “comprise”, “include”, and “contain” are used interchangeably and include not only closed definitions, but also semi-closed and open definitions. In other words, the terms include “consisting of”, “consisting essentially of”.
  • alkyl refers to a fully saturated cyclic or acyclic, branched or unbranched carbon chain moiety having the specified number of carbon atoms, or if not specified, up to 30 carbon atoms, such as 1 to 3, 1 to 6 carbon atoms.
  • alkyl having 1 to 8 carbon atoms refers to moieties such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl and octyl, and those moieties that are positional isomers of these moieties.
  • Alkyl having 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, docosyl, docosyl, triacontyl and tetracosyl, and the alkyl group may be substituted or unsubstituted.
  • C1-C6 alkyl or “C1-C3 alkyl” refers to a straight or branched chain alkyl group having 1-6 or 1-3 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.
  • alkylene refers to an alkyl group having a specified number of carbon atoms, such as 1 to 12 carbon atoms, which contains two points of connection to the rest of the compound on its longest carbon chain.
  • alkylene groups include C1-C6 alkylene groups, C1-C3 alkylene groups, specifically including methylene-(CH 2 )-, ethylene-(CH 2 CH 2 )-, n-propylene-(CH 2 CH 2 CH 2 )-, isopropylene-(CH 2 CH(CH 3 ))-, etc.
  • Alkylene groups can be cyclic or acyclic, branched or unbranched carbon chain moieties, and can be optionally substituted with one or more substituents.
  • C1-C6 alkoxy refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, or the like.
  • C2-C4 ester group refers to a group having a C1-C3 alkyl-OC(O)- structure or a group having an -OC(O) -C1-C3 alkyl structure, wherein the alkyl group may be linear or branched, for example CH3COO-, C2H5COO- , C3H8COO- , ( CH3 ) 2CHCOO- , -COOCH3 , -COOC2H5 , -COOC3H8 , or the like.
  • C2-C4 amide group refers to a group having a C1-C3 alkyl-CO-NH-structure or a group having a A group of the structure NH-CO-C1-C3 alkyl, wherein the alkyl group may be linear or branched, for example CH3 -CO-NH-, C2H5 - CO- NH-, C3H8-CO-NH-, -COOCH3, -CO-NH-C2H5 , -CO - NH - C3H8 , or the like.
  • C2-C4 acyl refers to a group having a C1-C3 alkyl-CO- structure, wherein the alkyl group may be linear or branched, such as CH3 -CO-, C2H5 - CO-, C3H8 - CO- , or the like.
  • C1-C3 haloalkyl refers to a linear or branched alkyl group having 1 to 3 carbon atoms in which one or more hydrogen atoms are substituted by a halogen group, such as monochloromethyl, dichloroethyl, trichloropropyl, or the like.
  • C1-C4 carboxyl refers to a group of a C1-C3 alkyl-COOH structure, wherein the alkyl group may be linear or branched, such as CH3COOH , C2H5COOH , C3H8COOH , ( CH3 ) 2CHCOOH , or the like.
  • C6-C10 aromatic ring refers to a monocyclic or bicyclic aromatic hydrocarbon group having 6 to 10 carbon atoms in the ring component, such as a benzene ring, a naphthalene ring, or a similar group.
  • aryl includes 3 to 12 substituted or unsubstituted monocyclic aromatic groups, wherein each atom of the ring is carbon (i.e., carbocyclic aromatic groups) or wherein one or more atoms are heteroatoms (i.e., heteroaryl groups).
  • aryl groups include 5- to 12-membered rings, more preferably 6- to 10-membered rings.
  • aryl also includes polycyclic ring systems having two or more rings, wherein two or more carbons are shared by two adjacent rings, wherein at least one ring is aromatic, for example, the other rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl and/or heterocyclic radicals.
  • Carbocyclic aromatic groups include benzene, naphthalene, phenanthrene, phenol, aniline, etc.
  • heteroaryl refers to an optionally substituted aromatic group, for example, a 5- to 7-membered monocyclic ring system having a ring containing at least one heteroatom and at least one carbon atom, such as pyrrolyl, thienyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, furan, imidazole, thiazole, oxazole, triazole, or the like.
  • cycloalkane ring refers to a non-aromatic group (including saturated, partially saturated or unsaturated groups) having 3 to 12 carbon atoms (e.g., 3 to 8, 4, 5, or 6 carbon atoms), having a monocyclic or condensed ring (including a bridged ring system and a spirocyclic system). Therefore, the alkane ring or alkane ring group can be a saturated alkane ring or an unsaturated alkane ring. In the condensed ring system of a saturated alkane ring, one or more rings are saturated alkane rings. In the condensed ring system of an unsaturated alkane ring, one or more rings can be a saturated alkane ring or an unsaturated alkane ring.
  • the term "5-8 membered heteroaromatic ring” refers to an aromatic heterocyclic ring system having one to multiple (preferably 1, 2 or 3) heteroatoms selected from N, O and S, and having 5-8 ring atoms. It should be understood that when multiple heteroatoms are contained, the heteroatoms may be the same, partially the same, or completely different.
  • the heteroaromatic ring is preferably a heteroaromatic ring containing 1 to 2 nitrogen atoms.
  • examples of 5-membered heteroaromatic rings include (but are not limited to): pyrrole ring, furan ring, thiophene ring, imidazole ring, pyrazole ring, oxazole ring, thiazole ring
  • examples of 6-membered heteroaromatic rings include (but are not limited to) pyridine ring, pyrazine ring, pyridazine ring, pyrimidine ring, or similar groups.
  • the term "5-8 membered heteroaryl” refers to an aromatic group having one to more (preferably 1, 2 or 3) heteroatoms selected from N, O and S, and having 5 or 8 ring atoms. It should be understood that when containing multiple heteroatoms, the heteroatoms may be the same, partially the same, or completely different.
  • examples of 5-membered heteroaryl include (but are not limited to): pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, or similar groups.
  • heteroaryl refers to 3-12 membered aromatic groups, which contain one or more heteroatoms selected from nitrogen, oxygen and sulfur, including monocyclic or polycyclic systems, and the polycyclic system can be a fused ring, a bridged ring system and a spirocyclic system.
  • fused heteroaromatic ring refers to a fused ring formed by the fusion of two or more aromatic groups, which contains one or more heteroatoms selected from nitrogen, oxygen and sulfur. Among them, one or more rings in the fused heteroaromatic ring have heteroatoms.
  • Heteroaryl includes substituted or unsubstituted aromatic 3-12 membered ring structures, more preferably 5-12 membered rings, more preferably 5-10 membered rings, and its rings
  • the structure includes 1-4 heteroatoms.
  • Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine.
  • Aryl and heteroaryl can be monocyclic, bicyclic or polycyclic. In this article, C5-C8 heteroaryl means that the number of ring atoms is 5-8.
  • heteroalkyl ring refers to a 3-12 membered non-aromatic group (including saturated, partially saturated or unsaturated groups) containing one or more heteroatoms selected from nitrogen, oxygen and sulfur, including a monocyclic or polycyclic system, and the polycyclic system can be a fused ring, a bridged ring system and a spirocyclic system. Therefore, the heteroalkyl ring or heteroalkyl ring group can be a saturated heteroalkyl ring or an unsaturated heteroalkyl ring. In a fused ring system, one or more rings can be an alkyl ring group, an aryl group or a heteroaryl group.
  • the number of ring atoms in the heteroalkyl ring can be 3 to 8, 4, 5 or 6.
  • the nitrogen atom and/or sulfur atom of the heterocyclic group is optionally oxidized to provide N-oxide, sulfinyl and sulfonyl moieties.
  • C3-C8 means that the number of ring atoms is 3-8.
  • the term "5-8 membered heteroalkyl ring” refers to any stable ring containing one or more (preferably 1, 2 or 3) heteroatoms selected from N, O and S, and a non-aromatic heterocyclic ring system with 5-7 ring atoms.
  • the heterocyclic ring may be a saturated, partially unsaturated, or unsaturated ring, but cannot be an aromatic ring.
  • the heteroalkyl ring is preferably a heteroalkyl ring containing 1 to 2 nitrogen atoms, which may optionally further include 1 or 2 oxygen atoms. It should be understood that when multiple heteroatoms are contained, the heteroatoms may be the same, partially the same, or completely different.
  • examples of 5-membered heteroalkyl rings include (but are not limited to) pyrrolidine rings, pyrroline rings, pyrazolidine rings, pyrazoline rings, 1,3-oxopentacyclic rings, and examples of 6-membered heteroalkyl rings include (but are not limited to) piperidine rings, morpholine rings, piperazine rings, 1,4-dioxane rings, or similar groups.
  • the term "four-membered heteroalkane ring” is any stable non-aromatic heterocyclic ring system containing one or more (preferably 1, 2 or 3) heteroatoms selected from N, O and S, and the number of ring atoms is 4.
  • the four-membered heteroalkane ring is preferably a four-membered heteroalkane ring containing 1 to 2 nitrogen atoms. It should be understood that when containing multiple heteroatoms, the heteroatoms can be the same, partially the same, or completely different. Examples of four-membered heteroalkane rings include (but are not limited to) oxetane, azetidine, or similar groups.
  • heterocyclic radical or “heterocyclic group” refers to a 3 to 12-membered ring structure, more preferably a 5 to 12-membered ring, more preferably a 5 to 10-membered ring, and its ring structure includes 1 to 4 heteroatoms.
  • the heterocycle can be a monocyclic, bicyclic, spirocyclic or polycyclic ring.
  • the heterocyclic radical includes, for example, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinoline, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenazine, phenothiazine, furan, phenoxazine, pyrrolidine, oxolane, thiopentane, oxazole, piperidine, piperazine, morpholine, lactone, lactams such as azetidinone and pyrrol
  • heterocycle may be substituted at one or more positions by substituents as described above, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, thiol, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, sulfamoyl, sulfinyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, -CF3 , -CN, etc.
  • substituents for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, amino, nitro, thiol, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carb
  • halogen refers to, for example but not limited to, radioactive and non-radioactive forms of fluorine, chlorine, bromine, iodine, etc. In a preferred embodiment, the halogen is selected from fluorine, chlorine and bromine.
  • halo refers to halogen substituted
  • autoimmune disease refers to myasthenia gravis, polymyositis, autoimmune myocarditis, polymyalgia rheumatica, psoriatic arthritis, rheumatoid arthritis, Sjögren's syndrome, ankylosing spondylitis, relapsing polychondritis, inflammatory bowel disease (such as Crohn's disease, ulcerative colitis), celiac disease, autoimmune hepatitis, Primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), immune-mediated kidney disease, interstitial cystitis, Addison's disease, autoimmune thyroid disease (e.g., Hashimoto's thyroiditis, Graves' disease), diabetes mellitus, dermatomyositis, psoriasis, alopecia areata, autoimmune or immune-mediated skin diseases, bullous pemphigoid, erythe
  • compositions and methods of administration are provided.
  • the compounds of the present invention have inhibitory activity and selectivity against integrins ⁇ v ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used to treat, prevent and alleviate pulmonary fibrosis.
  • the pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier.
  • safe and effective amount means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects.
  • the pharmaceutical composition contains 0.1-1000 mg of the compound of the present invention per dose, and more preferably, contains 0.5-500 mg of the compound of the present invention per dose.
  • the "one dose” is a capsule or tablet.
  • “Pharmaceutically acceptable carrier” refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility” here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds.
  • Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.
  • cellulose and its derivatives such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.
  • gelatin such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.
  • administrations include, but are not limited to, oral, rectal, parenteral (intravenous, intramuscular or subcutaneous), inhalation and topical administration.
  • parenteral intravenous, intramuscular or subcutaneous
  • inhalation topical administration.
  • a particularly preferred administration is oral.
  • Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.
  • the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrators, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example,
  • Solid dosage forms such as tablets, dragees, capsules, pills and granules may be prepared with coatings and shells, such as enteric coatings and other materials known in the art. They may contain opacifying agents and the active compound or compounds in such compositions may be The release of the active compound can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microcapsules with one or more of the above-mentioned excipients.
  • Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures.
  • the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.
  • an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottons
  • composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
  • Suspensions in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.
  • suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.
  • compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions.
  • Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.
  • Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants.
  • the active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.
  • the compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.
  • a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 0.2 to 1000 mg, preferably 0.5 to 500 mg.
  • the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.
  • Step 1 Dissolve 2-aminonicotinaldehyde (1 g, 8.2 mmol) and 1,1-dimethoxypropane-2-one (1.3 g, 10.6 mmol) in a 10/1 ethanol/water (22 ml) solution, slowly add sodium hydroxide solution (3 M, 3.6 ml) to the solution at 0°C under a nitrogen atmosphere, and stir the mixture at room temperature for 3 hours. Concentrate the reaction mixture, dissolve the residue in ethyl acetate (50 ml), and The organic phase was washed with saturated brine (25 ml*2), dried, filtered and concentrated to obtain 2 g of yellow solid 2-(dimethoxymethyl)-1,8-naphthyridine (A-1).
  • Step 2 A-1 (1.6 g, 7.8 mmol) was dissolved in ethanol (20 ml), platinum dioxide (50 mg, 0.2 mmol) was slowly added to the solution under nitrogen atmosphere, and the mixture was stirred at room temperature for 16 hours under hydrogen protection (15 psi). The reaction mixture was filtered, and the filtrate was concentrated to obtain 1 g of light yellow solid 7-(dimethoxymethyl)-1,2,3,4-tetrahydro-1,8-naphthyridine (A-2).
  • Step 3 A-2 (1.2 g, 5.8 mmol) was dissolved in water (10 ml), and 12 mmol/ml concentrated hydrochloric acid (1.1 ml, 13 mmol) was added to the solution at -5°C, and the mixture was stirred at 85°C for 2 hours. The mixture was cooled to room temperature, and the pH was adjusted to 11 by 3M sodium hydroxide solution. The aqueous phase was extracted with ethyl acetate (30 ml*2), and the organic phase was dried over Na 2 SO 4 , filtered and concentrated in vacuo to obtain a crude product.
  • Step 4 A-3 (3.1 g, 19.1 mmol) was dissolved in THF (60 ml), (Boc) 2 O (8.3 g, 38.2 mmol) and N, N-dimethylaminopyridine (4.7 g, 38.2 mmol) were added to the solution in sequence at room temperature, and the mixture was stirred at 80°C for 16 hours under nitrogen protection. The mixture was washed with saturated aqueous ammonium chloride solution (20 ml), extracted with ethyl acetate (20 ml*3), and the organic phase was dried over Na 2 SO 4 , filtered and concentrated in vacuo to obtain a crude product.
  • Step 5 A-4 (1 g, 3.8 mmol) was dissolved in THF (20 ml), sodium borohydride (173.1 mg, 4.6 mmol) was added at 0°C, and the mixture was stirred at 0°C for 1 hour under nitrogen protection. Water (5 ml) was added to the mixture to quench it, and ethyl acetate (10 ml*3) was extracted. The organic phase was washed with saturated brine, dried over Na 2 SO 4 , filtered and concentrated in vacuo to obtain a crude product.
  • Step 6 A-5 (600 mg, 2.3 mmol) was dissolved in DCM (10 ml), triphenylphosphine (893.1 mg, 3.4 mmol) and carbon tetrabromide (978.6 mg, 3 mmol) were added to the solution at room temperature, and the mixed liquid was stirred at room temperature for 3 hours under nitrogen protection.
  • Step 1 3-phenylacrylic acid (10 g, 67.5 mmol) was dissolved in thionyl chloride (50 ml, 689.3 mmol), the mixture was stirred at 90°C for 2 hours, and vacuum concentrated to obtain a residue. The residue was dissolved in DCM (200 ml), and sodium bicarbonate (10 g, 119. mmol) was added to the solution at room temperature to neutralize the excessive thionyl chloride. Then 4-dimethylaminopyridine (1 g, 8.2 mmol) and m-toluidine (7.2 g, 67.5 mmol) were added to the reaction solution in sequence and stirred for 14 hours.
  • the reaction solution was washed once with hydrochloric acid solution (5%, 150 ml) and sodium bicarbonate aqueous solution (150 ml), and the organic phase was dried over Na2SO4, filtered, and concentrated to obtain a crude product.
  • Step 2 Dissolve B-1 (15.7 g, 66 mmol) in chlorobenzene (200 ml), slowly add aluminum chloride (44 g, 330 mmol) under nitrogen atmosphere, and stir the reaction solution at 90°C for 2 hours under nitrogen protection. After cooling the reaction solution, pour it into ice water (300 ml), extract it with ethyl acetate (200 ml*2), wash the organic phase with saturated brine (100 ml), dry it with Na 2 SO 4 , filter it and concentrate it in vacuo to obtain a crude product.
  • Step 3 Dissolve B-2 (9.8 g, 61.4 mmol) in phosphorus oxychloride (130 ml, 1.4 mol), and stir the mixture at 100°C for 2 hours. Concentrate the mixture, slowly pour it into ice water (300 ml) and keep it in an ice bath, and add solid sodium hydroxide to adjust the pH to 7. The mixture is extracted with ethyl acetate (200 ml), and the organic phase is washed with saturated brine, dried over Na 2 SO 4 , and vacuum filtered and concentrated to obtain a crude product.
  • Step 5 Dissolve B-4 (570 mg, 1.3 mmol) in carbon tetrachloride (28 ml), add N-bromosuccinimide (258.3 mg, 1.5 mmol) and dibenzoyl peroxide (63.9 mg, 0.3 mmol) to the solution at room temperature, and stir the mixture at 80°C for 12 hours.
  • Step 1 Refer to the preparation method of intermediate A-5, filter and concentrate in vacuo to obtain 100 mg of white solid tert-butyl (6-(hydroxymethyl)pyridin-2-yl)carbamate (C-1).
  • Step 2 C-1 (100 mg, 445.9 ⁇ mol) and diisopropylethylamine (172.9 mg, 1.3 mmol) were dissolved in acetonitrile (4 ml), p-toluenesulfonyl chloride (47.2 mg, 668.9 ⁇ mol) was added at 0°C, and the mixture was stirred at room temperature for 3 hours under nitrogen protection.
  • Step 1 Dissolve 6-bromo-2-nitropyridine-3-ol (9 g, 41.1 mmol), ethyl bromoacetate (8.2 g, 49.3 mmol) and potassium carbonate (11.4 g, 82.2 mmol) in acetone (90 ml) in sequence and stir at 60°C for 2 hours under nitrogen protection. Wash the reaction solution with water (200 ml), extract with ethyl acetate (200 ml), wash the organic phase with saturated brine, dry over Na 2 SO 4 , filter and concentrate in vacuo to obtain 12 g of a brown oily crude product, ethyl 2-((6-bromo-2-nitropyridine-3-yl)oxy)acetate (D-1).
  • Step 2 Dissolve D-1 (12 g, 39.3 mmol), iron powder (11 g, 196.7 mmol) and ammonium chloride (10.5 g, 196.7 mmol) in 10/1 methanol/water (110 ml) in sequence, and stir at 60°C for 16 hours under nitrogen protection.
  • reaction solution was diluted with water (300 ml), extracted with ethyl acetate (300 ml), and the organic phase was washed with saturated brine, dried over Na 2 SO 4 , filtered and concentrated in vacuo to obtain a residue, which was dissolved in acetic acid (100 ml), stirred at 90°C for 3 hours, and then concentrated and filtered to obtain 7 g of a brown solid product 6-bromo-2H-pyrido[3,2-b][1,4]oxazine-3(4H)-one (D-2).
  • Step 3 Dissolve D-2 (1 g, 4.4 mmol) in THF (10 ml), add borane dimethyl sulfide (10 M, 1.1 ml) at room temperature, stir at 90°C for 1.5 hours, then add methanol (1 ml), stir at 90°C for 0.5 hours. Concentrate the reaction solution, wash with saturated sodium bicarbonate (50 ml), extract with ethyl acetate (50 ml), dry the organic phase over Na 2 SO 4 , filter and concentrate in vacuo to obtain 757 mg of 6-bromo-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazine (D-3) as a white solid.
  • Step 4 Dissolve D-3 (700 mg, 3.3 mmol), potassium vinyl trifluoroborate (654 mg, 4.9 mmol), palladium catalyst (119 mg, 0.2 mmol) and cesium carbonate (2.7 g, 8.1 mmol) in 1,4-dioxane (10 ml) in sequence and stir at 100 ° C for 2 hours under nitrogen protection.
  • Step 6 Dissolve D-5 (500 mg, 1.9 mmol) in 4/1 THF/H 2 O (12.5 ml), add sodium periodate (1.1 g, 5.3 mmol) and potassium osmate dihydrate (4.2 mg, 0.01 mmol) in turn at room temperature, and stir at room temperature for 1 hour.
  • the reaction solution was filtered, washed with water (30 ml), extracted with ethyl acetate (30 ml), and concentrated.
  • LC-MS ESI m/z: 289.2 [M+23] + .
  • Step 8 D-7 (50 mg, 0.2 mmol) was dissolved in DCM (3 ml), and carbon tetrabromide (93.4 mg, 0.3 mmol) and triphenylphosphine (74 mg, 0.3 mmol) were added successively at room temperature, and stirred at room temperature for 2 hours. The reaction solution was quenched by adding water (20 ml), extracted with DCM (20 ml), and the organic phase was dried, filtered, and concentrated.
  • Step 2 Refer to the preparation method of intermediate D-6 to obtain 900 mg of brown solid crude product 3-carbonyl-3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazine-6-carbaldehyde (E-2).
  • Step 2 Referring to the preparation method of intermediate D-4, the crude product was purified by normal phase silica gel column chromatography (EA) to obtain 160 mg of white solid product 5-vinyl-3H-imidazo[4,5-b]pyridine (F-2).
  • EA normal phase silica gel column chromatography
  • LC-MS: ESI m/z 246.4[M+H] + .
  • Step 4 Referring to the preparation method of intermediate D-6, 120 mg of light yellow solid product tert-butyl 5-formyl-3H-imidazo[4,5-b]pyridine-3-carboxylate (F) was obtained.
  • Step 1 (3-Bromophenyl)hydrazine hydrochloride (4.4 g, 19.7 mmol), potassium acetate (2.5 g, 25.6 mmol) and acetylacetone (2.1 g, 20.7 mmol) were dissolved in ethanol solution (20 ml), stirred at 85 ° C for 3 hours under nitrogen protection. Water was added to quench, extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over Na 2 SO 4 , filtered and concentrated in vacuo to obtain 5 g of 1-(3-bromophenyl)-3,5-dimethyl-1H-pyrazole (G-1) as a yellow oily product.
  • G-1 1-(3-bromophenyl)-3,5-dimethyl-1H-pyrazole
  • Step 2 G-1 (4 g, 15.9 mmol) and triisopropyl borate (1.5 eq.) were dissolved in anhydrous THF (40 ml), and n-butyl lithium (2.5 M, 16 ml) was slowly added dropwise at -65°C under nitrogen protection, and the mixture was stirred at -65°C to -60°C for 2 hours. Dilute hydrochloric acid (3 M, 11 ml) was added dropwise to the solution at -65°C to quench the reaction, and the pH was adjusted to 7 with sodium hydroxide solution (1 M), and EA was extracted.
  • Step 1 Dissolve 3,5-dibromoaniline (5 g, 19.9 mmol) in 40 ml of concentrated hydrochloric acid, add sodium nitrite (2.1 g, 29.9 mmol) to the solution under nitrogen atmosphere at 0°C, stir at 0°C for 1 hour, and then add stannous chloride (7.6 g,
  • Step 3 H-2 (500 mg, 1.5 mmol), bis-naphthalene boronate (462 mg, 1.8 mmol), potassium acetate (446 mg, 4.6 mmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (111 mg, 152 ⁇ mol) were dissolved in 5 ml of anhydrous Diox and stirred at 90°C for 2 hours under nitrogen atmosphere.
  • Step 4 H-3 (400 mg, 1.1 mmol) was dissolved in 4/1 THF/H 2 O (5 ml), sodium periodate (681 mg, 3.2 mmol) and 1M dilute hydrochloric acid (955 ⁇ l) were added to the reaction solution, and stirred at room temperature for 12 hours. The reaction solution was concentrated, and the crude product was separated by HPLC to obtain 70 mg of a white solid product (3-bromo-5-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)boronic acid (H). LC-MS: ESI m/z: 295.0 [M+H] + .
  • Step 3 Referring to the preparation method of intermediate H-1, the reaction solution was filtered and concentrated in vacuo to obtain 1 g of a yellow solid product (3-bromo-5-(tert-butyl)phenyl)hydrazine hydrochloride (I-3). LC-MS: ESI m/z: 243.1 [M+H] + .
  • Step 5 Referring to the preparation method of intermediate H-3, 500 mg of dark brown oily product 1-(3-(tert-butyl)-5-(4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-3,5-dimethyl-1H-pyrazole (I-5) was obtained.
  • Step 6 Referring to the preparation method of intermediate H, the crude product was separated by HPLC to obtain 55 mg of white solid (3-(tert-butyl)-5-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)boronic acid (I). LC-MS: ESI m/z: 273.2 [M+H] + .
  • Step 2 Dissolve J-1 (5 g, 34.7 mmol) in ethanol (50 ml) and add 10% palladium carbon (1.5 g), stir at room temperature under hydrogen atmosphere for 16 hours. Filter and concentrate the reaction solution in vacuo to obtain 5 g of 7-methyl-1,2,3,4-tetrahydro-1,8-naphthyridine (J-2) as a white solid product.
  • LC-MS: ESI m/z 249.4[M+H] + .
  • Step 4 J-3 (3.5 g, 14.1 mmol) and dimethyl carbonate (4.4 g, 49.3 mmol) were dissolved in THF (30 ml), and 2 M lithium diisopropylamine solution (10.7 ml) was added dropwise at -78 °C under nitrogen protection, and stirred for 1 hour.
  • LC-MS: ESI m/z 188.0 [M-Boc+1] + .
  • Step 2 L-1 (1.5 g, 6.3 mmol) and sodium hydride (506.1 mg, 12.7 mmol) were dissolved in THF (20 ml), stirred at 25°C for 0.5 hours, 2-(trimethylsilyl)ethoxymethyl chloride (1.6 g, 9.5 mmol) was slowly added dropwise to the solution at 0°C, and stirring was continued at 0°C for 2.5 hours. 15 ml of saturated NH 4 Cl solution was added at 0°C to quench the reaction, 20 ml of water was added to dilute, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over Na 2 SO 4 , filtered and concentrated in vacuo.
  • Step 3 Dissolve L-2 (1.2 g, 3.3 mmol), tetrahydroxydiborane (585.7 mg, 6.5 mmol), potassium acetate (961.8 mg, 9.8 mmol), 2-dicyclohexylphosphino-2'4'6-triisopropylbiphenyl (311.5 mg, 0.7 mmol) and methanesulfonic acid (2-dicyclohexylphosphino-2'4'6-triisopropyl-1,1'-biphenyl) (2'-amino-1,1'-biphenyl-2-yl) palladium (II) (276.5 mg, 0.3 mmol) in methanol (20 ml) and stir at 80°C for 3 hours under nitrogen protection. Concentrate the reaction solution and purify the crude product by HPLC to obtain 670 mg of a yellow solid product.
  • LC-MS: ESI m/z 334.6 [M+H] + .
  • Step 1 H-2 (1.2 g, 3.5 mmol), different cycloalkylboronic acids (1 equivalent), Pd(dppf)Cl 2 (0.1 equivalent), potassium carbonate (2 equivalents), dioxane/water solution (10:1, 7 ml), under nitrogen protection, heated at 93°C for 3 h; LCMS detected that the reaction was complete, quenched with water, and then ethyl acetate was added, and the yellow solid was removed by suction, extracted with ethyl acetate, and the organic phases were combined, dried over Na2SO4, filtered, and distilled under reduced pressure at 50°C to obtain a yellow oil. The crude product was purified by column chromatography (PE/EA ⁇ 20/1) to obtain the corresponding yellow oily product.
  • Step 1 Dissolve tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate (0.5 g, 2.4 mmol) in N,N-dimethylformamide (5 ml), add diisopropylethylamine (1. ml, 5.9 mmol) and methyl (E)-4-bromobut-2-enoate (506 mg, 2.8 mmol) at room temperature and stir for 2 hours.
  • Step 2 Dissolve tert-butyl (E)-6-(4-methoxy-4-carbonylbut-2-ene-1-yl)-2,6-diazaspiro[3.4]octane-2-carboxylate (0.6 g, 1.8 mmol) in DCM (5 ml), add trifluoroacetic acid (2 ml), and stir at room temperature for 2 hours. The reaction solution is directly spin-dried to obtain 550 mg of methyl (E)-4-(2,6-diazaspiro[3.4]octane-6-yl)but-2-enoate as a yellow oily product.
  • Step 3 Dissolve methyl (E)-4-(2,6-diazaspiro[3.4]octan-6-yl)but-2-enoate (0.6 g, 1.7 mmol), intermediate A-3 (330 mg, 2 mmol) and diisopropylethylamine (657 mg, 5.1 mmol) in 1,2-dichloroethane (5 ml) in sequence. and stirred at room temperature for 30 minutes. Then sodium cyanoborohydride (160 mg, 2.5 mmol) was added and the reaction was continued for 1.5 hours. The reaction solution was washed with water (30 ml) and extracted with ethyl acetate (30 ml*2).
  • Step 4 Methyl (E)-4-(2-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,6-diazaspiro[3.4]octan-6-yl)but-2-enoate (90 mg, 0.3 mmol), intermediate G (191 mg, 0.9 mmol), (1,5-cyclooctadiene)chlororhodium(I) dimer ([Rh(COD)Cl] 2 , 6.2 mg, 0.01 mmol), (R)-(+)-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl ((R)-(+)-BINAP, 15.7 mg, 0.03 mmol) and cesium carbonate (247 mg, 0.8 mmol) were dissolved in dioxane (2 ml) and water (0.2 ml) in sequence, heated to 95° C.
  • Step 1 Under nitrogen protection, at 0°C, tert-butyl 2,6-diazaspiro[3.4]octane-2-carboxylate (500 mg, 2.36 mmol, 1.0 eq) and TEA (595.7 mg, 5.9 mmol, 2.5 eq) were dissolved in DCM (5 mL), and CbzCl (803.5 mg, 4.7 mmol, 2.0 eq) was added dropwise. The reaction mixture was reacted for 16 h. The reaction solution was concentrated and the crude product was passed through a column.
  • Step 2 Under nitrogen protection, at 0°C, 6-benzyl 2-(tert-butyl) 2,6-diazaspiro[3.4]octane-2,6-dicarboxylate (295 mg, 0.9 mmol, 1.0 eq) was dissolved in DCM (6 mL), TFA (3 mL) was added dropwise, and the reaction was allowed to react for 2 h. The reaction solution was neutralized with ammonia water, and the reaction solution was concentrated to obtain 209 mg of crude yellow oily benzyl 2,6-diazaspiro[3.4]octane-6-carboxylate. LC-MS: ESI m/z: 247.1[M+H] + .
  • Step 3 Under nitrogen protection and room temperature, benzyl 2,6-diazaspiro[3.4]octane-6-carboxylate (209.7 mg, 0.85 mmol, 1.0 eq) and potassium carbonate (352.8 mg, 2.6 mmol, 3.0 eq) were dissolved in MeCN/DMF (2 mL/2 mL), and tert-butyl (E)-4-bromobut-2-enoate (178.7 mg, 0.8 mmol, 1 eq) was added dropwise at 65°C for 2 h. The reaction solution was concentrated, the crude product was dissolved in DCM, filtered, and the filtrate was concentrated.
  • Step 4 Under nitrogen protection and room temperature, benzyl (E)-2-(4-(tert-butyloxy)-4-carbonylbut-2-en-1-yl)-2,6-diazaspiro[3.4]octane-6-carboxylate (45 mg, 0.1 mmol, 1.0 eq), intermediate G (75.4 mg, 0.3 mmol, 3.0 eq), (1,5-cyclooctadiene)chlororhodium (I) dimer ([Rh(COD)Cl] 2 , 5.7 mg, 0.01 mmol, 0.1 eq), (R)-(+)-2,2′-bis(diphenylphosphino)-1,1′-binaphthyl ((R)-(+)-BINAP, 14.4 mg, 0.023 mmol, 0.2 eq) () and potassium hydroxide (19.6 mg, 0.3 mmol, 3.0 eq) in aqueous solution were dissolved
  • Step 5 Under nitrogen protection and room temperature, benzyl (S)-2-(4-(tert-butyloxy)-2-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-carbonylbutyl)-2,6-diazaspiro[3.4]octane-6-carboxylate (150 mg, 0.27 mmol, 1.0 eq) was dissolved in MeOH (2 mL), Pd/C (5 mg) and 1 drop of acetic acid were added, and the mixture was reacted at room temperature for 16 h. Celite was used for filtration, and the reaction solution was concentrated.
  • Step 6 Under nitrogen protection and room temperature, tert-butyl (S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(2,6-diazaspiro[3.4]octan-2-yl)butanoate (60 mg, 0.1 mmol, 1.0 eq) and potassium carbonate (58.4 mg, 0.4 mmol, 3.0 eq) were dissolved in DMF (6 mL), and a DMF solution of tert-butyl 7-(bromomethyl)-3,4-dihydro-1,8-naphthyridin-1(2H)-carboxylate (A) (30.5 mg, 0.13 mmol, 0.95 eq) was added dropwise at 65°C and reacted for 2 h.
  • Step 1 Dissolve tert-butyl 5-carbonyl-2,6-diazaspiro[3.4]octane-2-carboxylate (220 mg, 972.3 ⁇ mol) in THF (10 ml), add sodium hydride (110. mg, 2.8 mmol) and intermediate A (450 mg, 1.4 mmol) at 0°C, and stir at 60°C for 3 hours under nitrogen protection. Add saturated aqueous ammonium chloride solution (5 ml) to the reaction solution, and extract with ethyl acetate (15 ml*3). The organic phase is washed with saturated brine, dried over Na 2 SO 4 , filtered and concentrated in vacuo to obtain a crude product.
  • Step 2 Dissolve tert-butyl 7-((2-(tert-butoxycarbonyl)-5-carbonyl-2,6-diazaspiro[3.4]octan-6-yl)methyl)-3,4-dihydro-1,8-naphthyridin-1(2H)-carboxylate (220 mg, 465.5 ⁇ mol) in methanol (2 ml), add 4M hydrochloric acid dioxane (5 ml) at room temperature, and then stir at room temperature for 1 hour under nitrogen protection. LCMS detected that the raw material reacted completely.
  • reaction solution was concentrated to obtain 200 mg of a light yellow solid product 6-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,6-diazaspiro[3.4]octan-5-one.
  • Step 3 6-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,6-diazaspiro[3.4]octan-5-one (150 mg, 485.7 ⁇ mol) was dissolved in acetonitrile (5 ml), and methyl (E)-4-bromobut-2-enoate (34.78 mg, 194.3 ⁇ mol) and diisopropylethylamine (188.3 mg, 1.5 mmol) were added in sequence at room temperature, and stirred at 80°C for 16 hours under nitrogen protection. The reaction solution was concentrated to obtain a crude product.
  • Step 4 See the preparation method of intermediate 1-INTB step 4.
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 3.
  • Step 2 See the preparation method of intermediate 3-INTB step 2.
  • the reaction solution was concentrated to obtain 1 g of a yellow solid product 7-((2,7-diazaspiro[3.5]nonan-7-yl)methyl)-1,2,3,4-tetrahydro-1,8-naphthyridine.
  • LC-MS ESI m/z: 273.2 [M+H] + .
  • Step 3 Refer to the preparation method of intermediate 1-INT step 1.
  • Step 4 See the preparation method of intermediate 1-INTB step 4.
  • the reaction solution was concentrated and purified by high performance liquid chromatography to obtain 45 mg of reddish brown oily product.
  • 1 H NMR (400 MHz, METHANOL-d4) ⁇ ppm 7.57-7.41 (m, 2H) 7.39-7.26 (m, 3H) 6.72 (d, J 7.2 Hz, 1H) 6.04 (s, 1H) 4.11 (s, 2H) 3.98-3.78 (m, 2H) 3.65-3.58 (m, 2H) 3.55-3.50 (m, 3H) 3.48-3.40 (m, 3H) 3.35-3.28 (m, 3H) 3.15-3.08 (m, 3H) 2.83-2.73 (m, 3H) 2.72-2.64 (m, 1H) 2.22 (s, 3H) 2.20 (s, 3H) 2.12-1.95 (m, 4
  • Step 1 See the preparation method of intermediate 1-INT step 1.
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2. Obtain 810 mg of methyl (E)-4-(2,7-diazaspiro[3.5]nonan-7-yl)but-2-enoate as a pale yellow solid product.
  • Step 3 Dissolve intermediate A-3 (604.6 mg, 3.0 mmol) and TEA (628.7 mg, 6.2 mmol) in DCM (20 ml), add sodium acetate borohydride (987.5 mg, 4.7 mmol) and acetic acid (18.7 mg, 0.3 mmol) under nitrogen protection, and stir at room temperature overnight.
  • Step 4 See the preparation method of intermediate 1-INTB step 4.
  • Step 1 Dissolve tert-butyl 2,7-diazaspiro [3.5] nonane-7-carboxylate (500 mg, 2.2 mmol) and methyl (E)-4-bromobut-2-enoate (395.5 mg, 2.2 mmol) in 10 ml of acetonitrile, and then add potassium carbonate (610.7 mg, 4.4 mmol). The reaction solution is stirred at 80 ° C for 16 hours.
  • reaction solution is filtered and dried to obtain a crude product, which is purified by normal phase silica gel column chromatography (PE/EA from 100% to 40%) to obtain a yellow oily product tert-butyl (E)-2-(4-methoxy-4-carbonylbut-2-ene-1-yl)-2,7-diazaspiro [3.5] nonane-7-carboxylate 610 mg.
  • PE/EA normal phase silica gel column chromatography
  • Step 2 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by normal phase silica gel column chromatography (PE/EA from 100% to 20%) to obtain 940 mg of yellow oily product tert-butyl (S)-2-(2-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-methoxy-4-carbonylbutyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate.
  • LC-MS ESI m/z: 497.7 [M+H] + .
  • Step 3 Refer to the preparation method of intermediate 3-INTB in step 2.
  • the reaction solution was concentrated and dried to obtain 1 g of crude methyl (S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(2,7-diazaspiro[3.5]nonane-2-yl)butanoate (6-INTA).
  • LC-MS ESI m/z: 397.5 [M+H] + .
  • Step 3 Refer to the preparation method of intermediate 5-INTB in step 3.
  • the crude product was purified by normal phase silica gel column chromatography (DCM/MeOH from 100% to 90%) to obtain 30 mg of a yellow oily product.
  • Step 1 Refer to the preparation method of intermediate 3-INTB step 3.
  • the crude product was purified by normal phase silica gel column chromatography (PE/EA from 100% to 0%) to obtain 1 g of yellow oily product tert-butyl (E)-7-(4-methoxy-4-carbonylbut-2-en-1-yl)-2,7-diazaspiro[4.4]nonane-2-carboxylate.
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2.
  • the reaction solution was directly concentrated to obtain 1 g of white solid methyl (E)-4-(2,7-diazaspiro[4.4]nonane-2-yl)but-2-enoate.
  • Step 3 See the preparation method of intermediate 5-INTB step 3.
  • the crude product was separated by high performance liquid chromatography to obtain a yellow oily product methyl (E)-4-(7-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,7-diazaspiro[4.4]nonane-2-yl)but-2-ene Acid ester (7-INTA) 740mg.
  • Step 4 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by normal phase silica gel column chromatography (DCM/MeOH from 100% to 70%) to obtain 77 mg of a yellow oily product.
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 1.
  • the crude product was purified by normal phase silica gel column chromatography (PE/EA from 100% to 30%) to obtain 1 g of yellow oily product tert-butyl (E)-9-(4-methoxy-4-carbonylbut-2-en-1-yl)-3,9-diazaspiro[5.5]undecane-3-carboxylate.
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2. 500 mg of white solid methyl (E)-4-(3,9-diazaspiro[5.5]undec-3-yl)but-2-enoate was obtained.
  • Step 3 See the preparation method of intermediate 5-INTB step 3.
  • the crude product was separated by high performance liquid chromatography (formic acid) to obtain 60 mg of yellow oily product methyl (E)-4-(9-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-3,9-diazaspiro[5.5]undecane-3-yl)but-2-enoate (8-INTA).
  • Step 4 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by normal phase silica gel column chromatography (DCM/MeOH from 100% to 70%) to obtain 50 mg of a yellow oily product.
  • Step 1 Refer to the preparation method of intermediate 3-INTB step 3.
  • Step 3 See the preparation method of intermediate 5-INTB step 3.
  • the crude product was purified by HPLC to obtain 70 mg of a brown product, methyl (E)-4-(8-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,8-diazaspiro[4.5]decane-2-yl)but-2-enoate (9-INTA).
  • Step 4 See the preparation method of intermediate 1-INTB step 4.
  • Step 1 Refer to the preparation method of intermediate 3-INTB step 3.
  • Step 2 refers to the preparation method of intermediate 3-INTB step 2.
  • 120 mg of methyl (E)-4-(2,8-diazaspiro[4.5]decane-8-yl)but-2-enoate was obtained as a white solid product.
  • Step 3 See the preparation method of intermediate 5-INTB step 3.
  • the crude product was separated by high performance liquid chromatography to obtain 70 mg of yellow oily product methyl (E)-4-(2-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,8-diazaspiro[4.5]decane-8-yl)but-2-enoate (10-INTA).
  • Step 4 See the preparation method of intermediate 1-INTB step 4.
  • Step 1 See the preparation method of intermediate 5-INTB in step 3.
  • Step 2 Dissolve tert-butyl 1-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)spiro[indoline-3,4'-piperidine]-1'-carboxylate (140 mg, 0.32 mmol) in DCM (2 ml), add hydrochloric acid/methanol (2 ml, 4 M) to the reaction system, and stir at room temperature for 1 hour. The reaction solution is directly concentrated to obtain 80 mg of a yellow solid product 1-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)spiro[indoline-3,4'-piperidine].
  • Step 3 1-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)spiro[indoline-3,4'-piperidine] (40 mg, 0.15 mmol), methyl (E)-4-bromobut-2-enoate (18.73 mg, 0.10 mmol) and potassium carbonate (41 mg, 0.30 mmol) were added to DMF and stirred at 85°C for 4 hours. Water (10 ml) was added to the reaction solution and extracted with ethyl acetate (10 ml).
  • Step 4 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 1 Dissolve intermediate B (300 mg, 0.5 mmol) in acetonitrile (5 ml), add diisopropylethylamine (0.3 ml, 1.4 mmol) and intermediate 6-INTA (373.5 mg, 0.9 mmol) in turn at room temperature, heat the reaction solution to 80°C and stir for 6 hours.
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2. Obtain 230 mg of yellow solid product.
  • Step 1 See the preparation method of intermediate 12-INTB step 1.
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2. Obtain 100 mg of light yellow oily product.
  • Step 1 Dissolve intermediate D (30 mg, 0.01 mmol), intermediate 6-INTA (36.1 mg, 0.1 mmol) and N,N-diisopropylethylamine (35.3 mg, 0.3 mmol) in DCM (5 ml) in turn and stir at room temperature for 2 hours. The reaction solution was quenched with water (20 ml) and extracted with DCM (20 ml).
  • Step 2 Dissolve tert-butyl (S)-6-((2-(2-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-methoxy-4-carbonylbutyl)-2,7-diazaspiro[3.5]nonan-7-yl)methyl)-2,3-dihydro-4H-pyrido[3,2-b][1,4]oxazine-4-carboxylate (30 mg, 0.05 mmol) in DCM (5 ml), add methanolic hydrochloric acid (4 M, 5 ml) at room temperature, and stir for 1 hour. The reaction solution was directly spin-dried to obtain 25 mg of a yellow solid product.
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 1.
  • 1 H NMR 400 MHz, CDCl 3 -d) ⁇ 6.82 (m, 1H) 5.93 (m, 1H) 3.99 (s, 4H) 3.74 (s, 3H) 3.33 (s, 4H) 3.17 (m, 2H) 1.43 (s, 9H).
  • Step 2 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 3 Refer to the preparation method of intermediate 14-INTB step 2.
  • LC-MS: ESI m/z 369.6 [M+H] + .
  • Step 4 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 515.7 [M+H] + .
  • Step 1 Dissolve intermediate J (45 mg, 104 ⁇ mol) and 19-INTA (50.5 mg, 114.4 ⁇ mol) in acetonitrile (5 ml), then add diisopropylethylamine (40.3 mg, 312.1 ⁇ mol) and sodium iodide (31.2 mg, 208.1 ⁇ mol). The reaction solution was stirred at 60 ° C for 16 hours.
  • Step 1 Dissolve tert-butyl 4-formylpiperidine-1-carboxylate (5g, 23.4mmol) and 4-bromophenylhydrazine hydrochloride (5.2g, 23.4mmol) in chloroform (50ml) and ethanol (0.1ml). Cool to 0°C under nitrogen protection, slowly add trifluoroacetic acid (8g, 70.3mmol), and heat to 50°C after the addition is complete and stir for 16 hours. The reaction solution is quenched with water (30ml) and hydroxylamine aqueous solution (10ml), and extracted with DCM (30ml).
  • LC-MS:ESI m/z 365.4[M+H] + .
  • Step 2 Dissolve tert-butyl 5-bromospiro[indole-3,4'-piperidine]-1'-carboxylate (2.4 g, 6.6 mmol) in ethanol (30 ml), and slowly add sodium borohydride (1.2 g, 30.2 mmol) at room temperature. Stir at room temperature for 16 hours. The reaction solution was spin-dried, washed with water (50 ml), and extracted with DCM (50 ml).
  • Step 3 Dissolve tert-butyl 5-bromospiro[dihydroindole-3,4'-piperidine]-1'-carboxylate (750 mg, 2 mmol) and aldehyde in 1,2-dichloroethane (10 ml), add 2 drops of acetic acid at room temperature and stir for 30 minutes. Add sodium acetate borohydride (1.3 g, 6.1 mmol) and react for 16 hours.
  • LC-MS: ESI m/z 467.5[M+H] + .
  • Step 5 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 513.5 [M+H] + .
  • Step 1 See the preparation method of intermediate 14-INTB step 1.
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • LC-MS ESI m/z: 575.6 [M+H] + .
  • Step 2 Refer to the preparation method of intermediate 14-INTB step 2.
  • Step 3 Refer to the preparation method of intermediate 14-INTB in step 1.
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 2 Refer to the preparation method of intermediate 14-INTB step 2. 30 mg of methyl (S)-3-(3-(tert-butyl)-5-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(2,7-diazaspiro[3.5]non-2-yl)butanoate (25-INTA) was obtained as a yellow oily product. LC-MS: ESI m/z: 453.7 [M+H] + .
  • Step 3 Refer to the preparation method of intermediate 14-INTB in step 1.
  • Step 1 Add 3-bromo-4-methylpyridine (20 g, 116.3 mmol), palladium catalyst (5.3 g, 5.8 mmol), zinc cyanide (8.2 g, 69.8 mmol), zinc powder (0.8 g, 11.6 mmol) and DPPF (6.5 g, 11.6 mmol) to DMF (200 ml), replace nitrogen three times, and stir at 100 ° C for 16 hours under nitrogen atmosphere.
  • LC-MS: ESI m/z 119.0[M+H] + .
  • Step 2 Dissolve 4-methylnicotinonitrile (7.4 g, 62.6 mmol) and diethyl carbonate (74 g, 626.4 mmol) in THF (200 ml), replace with nitrogen three times, add NaH (12.5 g, 313.2 mmol) to the reaction system at 0°C, and stir at 60°C for 16 hours. Quench the reaction solution with saturated ammonium chloride (100 ml) and extract with ethyl acetate (100 ml).
  • Step 3 Add ethyl 2-(3-cyanopyridin-4-yl)acetate (5 g, 26.3 mmol) and Raney-Ni (5 g, 26.3 mmol) to ethanol (100 ml) and water (100 ml), replace with hydrogen three times, and heat to 50 ° C under hydrogen atmosphere and stir for 16 hours.
  • the reaction solution is filtered.
  • the filtrate is concentrated to obtain 3.8 g of yellow oily product 1,4-dihydro-2,7-diazinaphan-3(2H)-one.
  • LC-MS: ESI m/z 149.1[M+H] + .
  • Step 5 Dissolve 1,2,3,4-tetrahydro-2,7-naphthyridine (3.4 g, 25.3 mmol), (Boc) 2 O (8.3 g, 38 mmol), TEA (7.7 g, 76 mmol) and DMAP (309.6 mg, 2.5 mmol) in DCM (50 ml) and stir at room temperature for 16 hours. Quench the reaction solution with water (20 ml) and extract with DCM (20 ml).
  • Step 6 Dissolve tert-butyl 3,4-dihydro-2,7-naphthyridine-2(1H)-carboxylate (1.4g, 6.0mmol) and palladium hydroxide (557.6mg, 0.8mmol) in acetic acid (100ml), replace with hydrogen three times, heat to 80°C and stir for 12 hours under hydrogen atmosphere. Filter the reaction solution, add water (20ml) after concentrating the filtrate, adjust the pH to 9 with ammonia water, and extract with DCM (20ml).
  • Step 7 Dissolve tert-butyl octahydro-2,7-naphthyridine-2(1H)-carboxylate (800 mg, 3.3 mmol), potassium carbonate (1.4 g, 4.4 mmol) and methyl 4-bromocrotonate (595 mg, 3.3 mmol) in acetonitrile (15 ml) and stir at room temperature for 8 hours.
  • Step 8 Refer to the preparation method of intermediate 1-INTB in step 4.
  • Step 9 Refer to the preparation method of intermediate 14-INTB in step 2. 80 mg of yellow solid product methyl (3S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(octahydro-2,7-naphthyridin-2(1H)-yl)butanoate (29-INTA) was obtained.
  • Step 10 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 557.5 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 3-INTB step 3.
  • Step 2 Refer to the preparation method of intermediate 1-INTB step 4.
  • LC-MS ESI m/z: 483.3 [M+H] + .
  • Step 3 Refer to the preparation method of intermediate 14-INTB in step 2.
  • the reaction solution was directly concentrated to obtain 150 mg of yellow oily product methyl (S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(2,6-diazaspiro[3.4]octan-6-yl)butanoate (30-INTA).
  • Step 4 Refer to the preparation method of intermediate 5-INTB in step 3.
  • Step 1 Refer to the preparation method of step 1 of intermediate 12-INTB, wherein the preparation of 31-INTA refers to the preparation of 30-INTA.
  • Step 1 Refer to the preparation method of step 1 of intermediate 12-INTB, wherein the preparation of 32-INTA refers to the preparation of 30-INTA.
  • Step 1 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 513.1 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 513.6 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 527.7 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 1.
  • Step 2 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 3 Refer to the preparation method of intermediate 14-INTB in step 2.
  • the reaction solution was spin-dried to obtain 40 mg of a yellow oily product (S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(2,7-diazaspiro[3.5]nonanoic acid-7-yl)butanoic acid methyl ester (37-INTA).
  • Step 4 Refer to the preparation method of intermediate 5-INTB in step 3.
  • Step 1 Refer to the preparation method of step 3 of intermediate 5-INTB, wherein the preparation of 38-INTA refers to the preparation of 19-INTA.
  • LC-MS: ESI m/z 593.6 [M+H] + .
  • Step 1 Refer to the preparation method of step 3 of intermediate 5-INTB, wherein the preparation of 39-INTA refers to the preparation of 19-INTA.
  • LC-MS: ESI m/z 570.8 [M+H] + .
  • Step 1 See the preparation method of intermediate 5-INTB step 3.
  • Step 1 Refer to the preparation method of 5-INTB step 3.
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 3 See the preparation method of intermediate 5-INTB in step 3.
  • the crude product was purified by HPLC to obtain 15 mg of a colorless oily product.
  • LC-MS: ESI m/z 607.5 [M+H] + .
  • Step 1 See the preparation method of intermediate 1-INTB step 4.
  • Step 2 Refer to the preparation method of intermediate 2-INTB step 5.
  • the reaction solution was filtered and concentrated to obtain 520 mg of crude product methyl (S)-3-(3-(tert-butyl)-5-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(2,6-diazaspiro[3.4]octan-2-yl)butanoate (43-INTA).
  • LC-MS: ESI m/z 439.6[M+H] + .
  • Step 3 See the preparation method of intermediate 5-INTB in step 3.
  • the crude product was purified by HPLC to obtain 210 mg of a yellow oily product.
  • LC-MS: ESI m/z 585.5 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • LC-MS: ESI m/z 484.4[M+H] + .
  • Step 2 Dissolve tert-butyl (S)-2-(2-(5-bromopyridin-3-yl)-4-methoxy-4-carbonylbutyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate (65 mg, 134.7 ⁇ mol), 3,5-dimethyl-1H-pyrazole (19.4 mg, 202.1 ⁇ mol), methanesulfonic acid-2-(di-tert-butylphosphino)-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl(2-amino-1,1-biphenyl-2-yl)palladium(II) (11.5 mg) and cesium carbonate (109.8 mg, 336.9 ⁇ mol) in 1,4-dioxane (4 ml) in sequence, heat to 110°C under nitrogen protection and stir for 16 hours.
  • Step 4 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 544.3 [M+H] + .
  • Step 1 Refer to the preparation method of step 3 of intermediate 5-INTB, wherein the preparation of 24-INTA is the same as 6-INTA.
  • Step 1 Refer to the preparation method of step 3 of intermediate 5-INTB, wherein the preparation of 25-INTA is the same as 6-INTA.
  • Step 1 Refer to the preparation method of intermediate 5-INTB step 3.
  • Step 1 Refer to the preparation method of intermediate 5-INTB step 3.
  • LC-MS: ESI m/z 701.4 [M+H] + .
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2. Obtain 70 mg of light yellow oily product.
  • LC-MS: ESI m/z 601.4 [M+H] + .
  • Step 1 Refer to the preparation method of step 3 of intermediate 5-INTB, wherein the preparation of 49-INTA is the same as 30-INTA.
  • Step 1 Refer to the preparation method of step 3 of intermediate 5-INTB, wherein the preparation of 50-INTA is the same as 30-INTA.
  • Step 1 Refer to the preparation method of step 3 of intermediate 5-INTB, wherein the preparation of 51-INTA is the same as 32-INTA.
  • Step 1 Refer to the preparation method of intermediate 5-INTB step 3, wherein the preparation of 52-INTA is the same as 32-INTA.
  • LC-MS: ESI m/z 599.6 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 1 Dissolve tert-butyl 2,7-diazaspiro[4.4]nonane-2-carboxylate (100 mg, 441.9 ⁇ mol) in DCM (5 ml), then add TEA (184.5 ⁇ l, 1.3 mol) and methyl chloroacetate (62 mg, 574.4 ⁇ mol) in sequence, and react at room temperature for 2 hours.
  • reaction solution was spin-dried, washed with water (10 ml), extracted with ethyl acetate (10 ml), and the organic phase was dried and spin-dried to obtain 130 mg of yellow oily product tert-butyl 7-(2-methoxy-2-carbonylethyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate.
  • Step 3 Refer to the preparation method of intermediate 5-INTB in step 3.
  • the crude product was purified by HPLC to obtain 100 mg of a white solid product.
  • LC-MS: ESI m/z 345.5 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 3-INTB step 3.
  • Step 2 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 3 Refer to the preparation method of intermediate 3-INTB in step 2.
  • 500 mg of methyl (S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(1-oxa-4,9-diazaspiro[5.5]undec-4-yl)butanoate (63-INTA) was obtained as a light yellow oily product.
  • LC-MS: ESI m/z 427.6 [M+H] + .
  • Step 4 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 573.7 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 5-INTB step 3.
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2. Concentration afforded 30 mg of a light yellow oily product. LC-MS: ESI m/z: 528.6 [M+H] + .
  • Step 1 Dissolve tert-butyl 7-hydroxy-2-azaspiro[3.5]nonane-2-carboxylate (200 mg, 0.8 mmol), 1,8-naphthyridin-2-ol (1 equivalent) and triphenylphosphine (1.2 equivalents) in dry THF (20 ml) in sequence. Add di-tert-butyl azodicarboxylate (1.2 equivalents) dropwise to the solution at 0°C under nitrogen protection, and stir the reaction solution for 1 hour.
  • Step 2 Refer to the preparation method of intermediate J step 2. 50 mg of white solid product tert-butyl 7-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)oxy)-2-azaspiro[3.5]nonane-2-carboxylate was obtained. LC-MS: ESI m/z: 374.2 [M+H] + .
  • Step 3 Refer to the preparation method of intermediate 14-INTB in step 2. Obtain 40 mg of white solid product 7-((2-azaspiro[3.5]nonan-7-yl)oxy)-1,2,3,4-tetrahydro-1,8-naphthyridine. LC-MS: ESI m/z: 274.2 [M+H] + .
  • Step 4 Refer to the preparation method of intermediate 3-INTB in step 3.
  • Step 5 Refer to the preparation method of intermediate 1-INTB in step 4.
  • the crude product was purified by HPLC to obtain 26 mg of a yellow solid product.
  • Step 1 Refer to the preparation method of intermediate 3-INTB step 3.
  • Step 2 Refer to the preparation method of intermediate 14-INTB step 2. 640 mg of methyl (E)-4-((7-azaspiro[3.5]nonan-2-yl)amino)but-2-enoate was obtained as a yellow solid product.
  • Step 3 See the preparation method of intermediate 5-INTB step 3.
  • the crude product was subjected to high performance liquid chromatography to obtain a brown solid product methyl (E)-4-((7-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-7-azaspiro[3.5]nonane-2-yl)amino)butane-2-yl Enoic acid ester (66-INTA) 250 mg.
  • LC-MS: ESI m/z 385.5 [M+H] + .
  • Step 4 See the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 50 mg of a brown solid product.
  • LC-MS: ESI m/z 557.4 [M+H] + ;
  • Step 1 Refer to the preparation method of intermediate 3-INTB step 3.
  • Step 2 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 3 Refer to the preparation method of intermediate 14-INTB in step 2. 110 mg of methyl (S)-4-(8,8-difluoro-2,6-diazaspiro[3.4]octan-6-yl)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)butanoate (67-INTA) was obtained as a light yellow oily product.
  • LC-MS: ESI m/z 419.4 [M+H] + .
  • Step 4 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 565.5 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 409.5 [M+H] + .
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2. 500 mg of a light yellow oily product 7-((5,5-difluoro-2,7-diazaspiro[3.5]nonan-7-yl)methyl)-1,2,3,4-tetrahydro-1,8-naphthyridine was obtained.
  • Step 3 See the preparation method of intermediate 3-INTB step 3.
  • Step 4 See the preparation method of intermediate 1-INTB step 4.
  • LC-MS: ESI m/z 579.6 [M+H] + .
  • Step 1 Refer to the preparation method of step 4 of intermediate 1-INTB, wherein the preparation of 72-INTA is the same as 71-INTA.
  • LC-MS: ESI m/z 561.3 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 110 mg of a yellow oily product.
  • LC-MS: ESI m/z 527.4 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 210 mg of colorless oily product tert-butyl (S)-2-(2-(3-bromo-5-(tert-butyl)phenyl)-4-methoxy-4-carbonylbutyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate.
  • Step 2 Refer to the preparation method of intermediate 14-INTB step 2.
  • the reaction solution was directly concentrated to obtain 75 mg of yellow oily product methyl (S)-3-(3-bromo-5-(tert-butyl)phenyl)-4-(2,7-diazaspiro[3.5]nonan-2-yl)butanoate (76-INTA).
  • Step 3 See the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 583.6 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • LC-MS: ESI m/z 500.6 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 180 mg of yellow solid product tert-butyl (S)-2-(4-methoxy-2-(3-(2-methoxyethoxy)phenyl)-4-carbonylbutyl)-2,7-diazaspiro[3.5]nonane-7-carboxylate.
  • Step 3 See the preparation method of intermediate 5-INTB step 3.
  • the crude product was purified by HPLC to obtain 90 mg of a yellow oily product.
  • LC-MS: ESI m/z 523.5 [M+H] + ; 1 H NMR (400 MHz, METHANOL-d 4 ) 8.42 (s, 2H), 7.34-7.24 (m, 2H), 6.94-6.85 (m, 3H), 6.59-6.52 (m, 1H), 5.28-5.05 (m, 4H), 4.17-4.11 (m, 2H), 3.78-3.75 (m, 2H), 3.72 (s, 2H), 3.61 (s, 3H), 3.57-3.53 (m, 4H), 3.44 (s, 3H), 3.44-3.42 (m, 1H), 2.82-2.61 (m, 8H), 1.97-1.87 (m, 6H).
  • Step 1 Refer to the preparation method of intermediate 1-ITNB step 4.
  • Step 2 Dissolve tert-butyl (S)-2-(2-(5-bromo-2-fluorophenyl)-4-methoxy-4-carbonylbutyl)-2,6-diazaspiro[3.4]octane-6-carboxylate (380 mg, 0.8 mmol), (5-bromo-2-fluorophenyl)boric acid (90.3 mg, 0.9 mmol), palladium catalyst (66.9 mg, 0.1 mmol) and cesium carbonate (765.2 mg, 2.4 mmol) in dioxane (10 ml), replace with nitrogen three times, and add under nitrogen protection. Heat to 120°C and stir for 16 hours.
  • Step 3 Refer to the preparation method of intermediate 3-INTB in step 2.
  • the crude product was purified by HPLC to obtain 60 mg of methyl (S)-3-(5-(3,5-dimethyl-1H-pyrazol-1-yl)-2-fluorophenyl)-4-(2,6-diazaspiro[3.4]octan-2-yl)butanoate (80-INTA) as a yellow oily product.
  • LC-MS: ESI m/z 401.5[M+H] + .
  • Step 4 Refer to the preparation method of intermediate 5-INTB in step 3.
  • the crude product was purified by HPLC to obtain 40 mg of a yellow oily product.
  • LC-MS: ESI m/z 547.4 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 3 See the preparation method of intermediate 5-INTB step 3.
  • the crude product was purified by high performance liquid chromatography to obtain 80 mg of colorless oily product tert-butyl (S)-2-(3-(4-methoxy-4-carbonyl-1-(6-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,6-diazaspiro[3.4]octan-2-yl)butan-2-yl)phenyl)-1H-pyrrole-1-carboxylate.
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • Step 2 Dissolve benzyl (S)-2-(2-(3-bromo-5-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-methoxy-4-carbonylbutyl)-2,6-diazaspiro[3.4]octane-6-carboxylate (400 mg, 0.7 mmol), cyclopropylboronic acid (87 mg, 1 mmol), cesium carbonate (547 mg, 1.7 mmol) and palladium catalyst (49 mg, 0.1 mmol) in dioxane (10 ml) in sequence. Heat to 100°C and stir for 3 hours under nitrogen protection.
  • Step 4 See the preparation method of intermediate 1-INTB in step 3.
  • LC-MS: ESI m/z 569.4 [M+H] + .
  • Step 1 See the preparation method of intermediate 1-INTB step 4.
  • LC-MS: ESI m/z 357.3[M+H] + .
  • Step 2 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 21 mg of a colorless oily product.
  • LC-MS: ESI m/z 509.6 [M+H] + .
  • Step 1 See the preparation method of intermediate 20-INTB step 1.
  • Step 2 See the preparation method of intermediate 20-INTB step 2.
  • the crude product was purified by HPLC to obtain 50 mg of a yellow solid product.
  • LC-MS: ESI m/z 543.5 [M+H] + .
  • the intermediate 43-INTA was used as the starting material, and the preparation method and steps were the same as those for 90-INTB to obtain 400 mg of a yellow oily product.
  • LC-MS: ESI m/z 599.6 [M+H] + .
  • the intermediate 82-INTA was used as the starting material, and the preparation method and steps were the same as those for 90-INTB to obtain 350 mg of a yellow oily product.
  • LC-MS: ESI m/z 583.4 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 3-INTB step 3.
  • Step 2 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by normal phase silica gel column chromatography (EA) to obtain 140 mg of yellow oily product tert-butyl 7-((S)-2-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-methoxy-4-carbonylbutyl)-1,7-diazaspiro[4.4]nonane-1-carboxylate.
  • Step 4 Refer to the preparation method of intermediate 5-INTB in step 3.
  • LC-MS: ESI m/z 543.6 [M+H] + .
  • Step 1 Dissolve pyridine-2,6-diamine (5g, 45.8mmol) and 1,1,3,3-tetramethoxypropane (7.5g, 45.8mmol) in phosphoric acid (50ml), heat to 70°C under nitrogen protection, and stir for 2 hours. Adjust the pH of the reaction solution to 10 with 5M sodium hydroxide aqueous solution at 0°C, filter and rinse the filter cake with DCM (50ml*3), continue to extract the obtained solution with DCM (200ml*3), wash the organic phase with saturated brine (100ml*2), dry with Na 2 SO 4 , filter, and spin dry to obtain a crude product.
  • phosphoric acid 50ml
  • Step 2 Referring to the preparation method of intermediate A-2, 290 mg of crude product 5,6,7,8-tetrahydro-1,8-naphthyridin-2-amine was obtained.
  • LC-MS: ESI m/z 151.4 [M+H] + .
  • Step 4 Refer to the preparation method of intermediate 6-INTB in step 1.
  • LC-MS: ESI m/z 312.3 [M+H] + .
  • Step 5 Refer to the preparation method of intermediate 1-INTB in step 4.
  • LC-MS: ESI m/z 481.6 [M+H] + .
  • LC-MS: ES m/z 452.4[M+Na] + .
  • Step 7 Referring to the preparation method of intermediate D-6, the crude product was purified by HPLC to obtain 70 mg of yellow oily product methyl (S)-3-(3-(3,5-dimethyl-1H-pyrazol-1-yl)phenyl)-4-(7-formyl-3,4-dihydroisoquinolin-2(1H)-yl)butanoate (95-INTA).
  • Step 8 Refer to the preparation method of intermediate 1-INTB in step 3.
  • the crude product was purified by HPLC to obtain 10 mg of a yellow solid product.
  • LC-MS: ESI m/z 565.4 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 10 mg of a yellow solid product.
  • LC-MS: ESI m/z 635.4 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 10 mg of a yellow solid product.
  • LC-MS: ESI m/z 605.2 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 10 mg of a yellow solid product.
  • LC-MS: ESI m/z 563.2 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 10 mg of a yellow solid product.
  • LC-MS: ESI m/z 581.2 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 10 mg of a yellow solid product.
  • LC-MS: ESI m/z 559.3 [M+H] + .
  • Step 1 See the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 50 mg of methyl (S)-3-(5-bromopyridin-3-yl)-4-(5,5-difluoro-7-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,7-diazaspiro[3.5]nonane-2-yl)butanoate as a yellow solid product.
  • Step 2 See the preparation method of intermediate 44-INTB step 2.
  • the crude product was purified by HPLC to obtain 15 mg of a yellow solid product.
  • LC-MS: ESI m/z 580.3 [M+H] + .
  • Step 2 Refer to the preparation method of intermediate 3-INTB step 2. Obtain yellow oily product 7-(2-(5,5-difluoro-2,7-diazaspiro[3.5]nonan-7-yl)ethyl)-1,2,3,4-tetrahydro-1,8-naphthyridine.
  • LC-MS: ESI m/z 323.3 [M+H] + .
  • Step 3 Refer to the preparation method of intermediate 3-INTB step 3.
  • LC-MS: ESI m/z 421.2 [M+H] + .
  • Step 4 Refer to the preparation method of intermediate 1-INTB step 4. A pale yellow solid product is obtained.
  • LC-MS: ESI m/z 593.3 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 50 mg of a yellow solid product.
  • LC-MS: ESI m/z 617.4 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 50 mg of yellow solid product methyl (S)-3-(5-bromo-2-fluorophenyl)-4-(8,8-difluoro-6-((5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)methyl)-2,6-diazaspiro[3.4]octan-2-yl)butanoate.
  • Step 2 See the preparation method of intermediate 80-INTB step 2.
  • LC-MS: ESI m/z 583.3 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 50 mg of a yellow solid product.
  • LC-MS: ESI m/z 621.4 [M+H] + .
  • Step 1 Refer to the preparation method of intermediate 1-INTB step 4.
  • the crude product was purified by HPLC to obtain 50 mg of a yellow solid product.
  • LC-MS: ESI m/z 605.4 [M+H] + .
  • Step 1 Refer to the preparation of intermediate 1-INTB in step 4.
  • the crude product was purified by HPLC to obtain 50 mg of a yellow solid product.
  • LC-MS: ESI m/z 561.3 [M+H] + .
  • Step 1 Using tert-butyl (E)-8,8-difluoro-2-(4-methoxy-4-carbonylbut-2-en-1-yl)-2,6-diazaspiro[3.4]octane-6-carboxylate as the starting material, refer to the preparation method of intermediate 1-INTB step 4.
  • the principle is to detect the change in molecular weight of the fluorescently labeled small molecule before and after the interaction with other molecules, and calculate the fluorescence polarization values in the horizontal and vertical directions for correlation analysis. If the binding equilibrium between the fluorescently labeled small molecule and the macromolecule is established, it moves slowly when excited, and the measured fluorescence polarization light value will increase. If the binding between the fluorescently labeled small molecule and the macromolecule is replaced by other ligands, its rotation or flipping speed in the free state will become faster, the emitted light will be depolarized relative to the excitation light plane, and the measured polarization light value will decrease, thereby calculating the fluorescence anisotropy of the sample.
  • the experimental reaction system was 40ul, and duplicate wells were set up.
  • the buffer for ⁇ V ⁇ 6 was: 50mM HEPES PH7.4, 150mM Nacl, 0.5mM CHAPS, 0.4mM MgCl2, and the buffer for ⁇ V ⁇ 1 and ⁇ V ⁇ 8 was: 50mM HEPES PH7.4, 150mM Nacl, 0.5mM CHAPS, 0.1mM MnCl2.
  • the working concentration of the fluorescent substrate is 1nM
  • the working concentrations of ⁇ V ⁇ 6 (recombinant human integrin produced by R&D systems, catalog number 3817-AV) and ⁇ V ⁇ 8 (recombinant human integrin produced by R&D systems, catalog number 4135-AV) are both 8nM
  • the working concentration of ⁇ V ⁇ 1 (catalog number 6579-AVB) is 12.5nM
  • the initial screening concentrations of the compound are 1uM and 100nM
  • the final concentration of DMSO is 0.2%. All components are mixed in a 384-well plate (corning catalog number: CLS3575) and reacted at room temperature (about 24°C) for 2 hours before determining the Anisotropy value.
  • the testing instrument is the BioTek brand SYNERGY neo2 microplate reader, with an Excitation of 485nM and an emission of 530nM.
  • the wells with only buffer added are used as blank controls for system readings.
  • the inhibition rate is calculated by taking the average value of the duplicate wells, and the IC 50 value of the compound with an inhibition rate greater than 50% under the condition of 100nM of the compound is determined.
  • PLN-74809 and GSK-3008348 were selected as control compounds in this experiment.
  • PLN-74809 (CAS: 2376257-44-0) and GSK-3008348 (CAS: 1629249-33-7) are two very potential integrin ligand molecules.
  • the experimental treatment method is the same as above.
  • C is the Anisotropy value of complete binding of the fluorescent substrate to the protein
  • F is the Anisotropy value at the corresponding concentration of the compound
  • B is the background value of the Anisotropy of the fluorescent substrate.
  • the S curve is drawn with the compound concentration and the corresponding inhibition rate value (Table 1), and the corresponding IC 50 value is calculated (Table 2).
  • the a-level embodiment has excellent inhibitory activity against the target subtype; the b-level embodiment has a strong inhibitory activity; the c-level embodiment has a general inhibitory activity.
  • the A-level embodiment has strong selectivity for subtype 2 compared with subtype 1; the B-level embodiment has moderate selectivity; and the C-level has no selectivity.
  • IC50 IC50 ⁇ 100 nM
  • c IC50 > 1000 nM
  • B: Fold 1.0-4.0, indicating that the compound has moderate selectivity for a certain subtype
  • N/A means no test was performed;
  • the examples show different inhibitory effects on one, two or three subtypes of ⁇ v ⁇ 1, ⁇ v ⁇ 6 and ⁇ v ⁇ 8, respectively, and especially show excellent inhibitory effects and selectivity on the single subtype of ⁇ v ⁇ 6 and the two subtypes of ⁇ v ⁇ 1 and ⁇ v ⁇ 6, such as Examples 43, 44, 71, 72, 96-103 (aaaCA type) and the like have excellent inhibitory effects on both ⁇ v ⁇ 1 and ⁇ v ⁇ 6 and good selectivity for ⁇ v ⁇ 8, and the selectivity is much better than the positive reference PLN-74809.
  • Examples 1, 5, 49-50, 52-54, 75, 77-78 and the like have excellent inhibitory activity and specific selectivity for ⁇ v ⁇ 6.
  • Fibroblasts are the main effector cells in fibrotic diseases. When activated, they can undergo functional and phenotypic changes and transdifferentiate into myofibroblasts (MFb), which have contractile potential and strong collagen synthesis ability. This is the main cellular process that induces fibrotic lesions.
  • the phenotypic characteristics of activated cells are the expression of ⁇ -smooth muscle actin ( ⁇ -Smooth muscle actin, ⁇ -SMA) and fibronectin (Fibronectin, FN), so they can be activated by these two proteins.
  • the changes in protein expression levels were used to verify the inhibitory effect of integrin ligands on fibroblast activation.
  • Cell line human fetal lung fibroblasts (HFL-1); TGF- ⁇ : R&D; culture medium: F12K culture medium from Hyclone; serum: Gibco; Fibronectin antibody: abcam, catalog number: ab45688; ⁇ -SMA antibody: abcam, catalog number: ab124964.
  • HFL-1 cells were inoculated in F12K medium containing 10% FBS and cultured routinely. When treated with different compounds, 1 ⁇ 106 cells/well were inoculated in 6-well plates, cultured under normal culture conditions for 24 hours after inoculation, and then replaced with serum-free medium. The excipient DMSO and different concentrations of compounds (3 concentrations for each compound, 3 replicates for each concentration) were pretreated for 2 hours, and then 10 ng/ml of TGF- ⁇ recombinant protein was added for 24 hours. After treatment, the cells were recovered for protein immunoblotting (Western blotting, WB) detection of Fibronectin and ⁇ -SMA.
  • WB Western blotting
  • Pirfenidone and GSK-3008348 were selected as control compounds for this experiment.
  • Pirfenidone (CAS: 53179-13-8) is a widely used anti-fibrotic drug
  • GSK-3008348 (CAS: 1629249-33-7) is a highly potential integrin ligand molecule developed by GlaxoSmithKline (GSK Plc.).
  • the drug concentration of pirfenidone was 0.5 mg/mL, and the drug concentration of GSK-3008348 was the same as that of the test compound (3 concentrations, 3 replicates for each concentration), and the treatment method was the same as above.
  • Protein extraction Wash the cells with PBS, add 100ul RIPA lysis buffer to each well, scrape the cells with a cell scraper and collect them in an EP tube, place on ice for 30 minutes, centrifuge at 13000g for 10 minutes after sufficient lysis, and take the supernatant and place it in a new EP tube. Protein quantification is performed according to the instructions of the BCA protein concentration assay kit (enhanced). According to the quantitative results, dilute the sample to make the protein concentration consistent. Add the loading buffer and mix well, heat in a boiling water bath for 3-5 minutes to fully denature the protein, and load the sample after cooling to room temperature.
  • Make gel inject separation gel, leave 1/5 of the space, and fill it up with deionized water. After standing at room temperature for 1 hour, pour out the deionized water, dry it with absorbent filter paper, and then fill it up with concentrated gel; then quickly insert the comb, and pay attention to remove bubbles with the remaining concentrated gel, and then stand at room temperature for 30 minutes.
  • Sample loading/electrophoresis/transfer The total protein loading amount of each sample is 25ug, which is added to the sample well with a sample gun.
  • Electrophoresis Run at 70V to the lower layer of the concentrated gel and the upper layer of the separation gel, and then run at 120V to the end.
  • Transfer First soak the 0.45um PVDF membrane, filter paper, and sponge in the transfer buffer, and mark the PVDF membrane; after the electrophoresis is completed, select the appropriate gel; make a transfer sandwich, put it on the rack, fill it with electrotransfer buffer, and run at a constant current of 230mA for 120min.
  • Antigen blocking and antibody incubation First wash the membrane with PBS, add Western blocking solution (5% nonfat-milk), and incubate on a shaker at room temperature for 1 hour.
  • Primary antibody incubation Add primary antibodies ⁇ -SMA, Fibronectin and internal reference Actin respectively, overnight at 4°C; wash the primary antibody, add TBST, and wash 3 times on a shaker at room temperature, 5 minutes each time.
  • Secondary antibody incubation Add the corresponding secondary antibody and incubate at room temperature with shaking for 1 hour. Wash the secondary antibody, add TBST, and wash 4 times on a shaker at room temperature, 6 minutes each time. Finally, the fluorescence imager is used for detection, and the optical density value of the protein band is quantitatively read. The optical density value of the actin band is used as the internal reference to compare the expression of each target protein.
  • this experiment uses the Antibody Stripping method, first detecting the ⁇ -SMA protein, then using the stripping buffer to remove the ⁇ -SMA primary antibody, and then detecting the Actin protein.
  • the incubation methods for the two antibodies are the same as above.
  • Embodiments 1, 2, 7, 15, and 25 have significant promoting degradation effects on fibrosis proteins ⁇ -SMA and Fibronectin (as shown in Figure 1).
  • Embodiments 2, 7, 15, and 25 significantly reduce the expression of ⁇ -SMA protein, have an excellent promoting degradation effect on fibrosis protein ⁇ -SMA, and are significantly better than the positive references pirfenidone and GSK-3008348, showing excellent anti-fibrosis effects
  • Embodiments 1, 2, 7, and 25 significantly reduce the expression of Fibronectin protein, have an excellent promoting degradation effect on fibrosis protein Fibronectin, and are also better than the positive references pirfenidone and GSK-3008348.
  • Embodiments 2, 7, and 25 all have excellent promoting degradation effects on both ⁇ -SMA and Fibronectin proteins, and the effects are significantly better than the positive references GSK-3008348 and pirfenidone. This indicates that the ligands of integrins ⁇ v ⁇ 1, 6, and 8 in the present invention have a significant inhibitory effect on the secretion of ⁇ -SMA protein and Fibronectin protein as well as the activation of fibroblasts, and have the potential to become anti-fibrosis drugs.
  • Substrate, liver microsomes, MgCl 2 , potassium phosphate buffer, and water were added to a 96-well plate and gently vortexed to mix.
  • the microsome mixture was pre-incubated in a 37°C water bath for 5 min, and the reaction was initiated by adding the NADPH generating system. After incubation for 0, 10, 20, and 30 min, 40 ⁇ L of the reaction mixture was mixed with 100 ⁇ L of acetonitrile (with internal standard 100 ng/mL) to terminate the reaction. After the incubation, the mixture was centrifuged at 4600 rpm for 10 min at 4°C. 60 ⁇ L of the supernatant was taken, diluted 1:1 with water, and 5 ⁇ L was injected for LC-MS/MS analysis.
  • a Waters XBridge C18 column (50 mm ⁇ 2.1 mm, 5 ⁇ m) was used with an injection volume of 5 ⁇ L. The flow rate was 1 mL/min and the running time was 1.5 min. The mobile phase A was water containing 0.1% formic acid, and B was acetonitrile containing 0.1% formic acid.
  • the gradient elution program is shown in Table 4 below:
  • LC-MS/MS High performance liquid chromatography tandem mass spectrometry
  • model API 4500 (AB Sciex).
  • Electrospray ionization (ESI) conditions and multiple reaction monitoring (MRM) mode were used to detect the eluted compounds.
  • the dry gas temperature was 450°C
  • the pressure was 50 psi
  • the nebulizing gas pressure was 20 psi.
  • the compounds of the present invention are metabolically stable in the liver microsomes of humans, mice, rats, dogs and monkeys, among which Examples 1, 2, 6, 72 and 73 have better half-lives in human liver microsomes than the positive reference PLN-74809, and Examples 2, 6, 73 and 117 have better half-lives in dog liver microsomes than PLN-74809.
  • Other compounds of the present invention also have similar physicochemical properties.
  • mice Male SD rats, weighing about 200 g, were purchased from Shanghai Xipuer-Bikai Experimental Animal Co., Ltd., with 3 rats in each group. Each test compound was administered orally (PO) and intravenously (IV).
  • the injection dose was 1 mg/kg, and the test compound was prepared into a 0.1 mg/mL solution using the same solvent and injected into the tail vein at a volume of 10 mL/kg. Blood was collected at 5, 15, 30, 60, 120, 240, 480, 720, and 1440 minutes after administration to determine the blood drug concentration.
  • the oral dose was 10 mg/kg.
  • Each blood collection volume was about 200 ⁇ L, stored in an EDTA-2K anticoagulation tube, and centrifuged at 5500 rpm for 10 min to separate plasma. Take 20 ⁇ L of the test plasma sample, standard plasma sample, and QC plasma sample at each time point, mix them with 100 ⁇ L acetonitrile (with internal standard 100 ng/mL), shake at room temperature for 10 min, and then centrifuge at 3700 rpm for 18 min at 4 ° C. Take 60 ⁇ L of the supernatant, dilute it 1:1 with water, shake it at room temperature for 10 min, and then inject 5 ⁇ L for LC-MS/MS analysis.
  • LC-MS/MS High performance liquid chromatography tandem mass spectrometry
  • model API 4500 (AB Sciex)
  • Electrospray ionization (ESI) conditions and multiple reaction monitoring (MRM) mode were used to detect the eluted compounds.
  • the ion source temperature was 450°C
  • the spray gas and auxiliary heating gas pressures were both 50 psi
  • the curtain gas pressure was 20 psi
  • the collision chamber outlet voltage was 13.0 V.
  • the declustering voltages of the internal standard and the test compound were 79, 130, and 135 eV, respectively, and the collision energies were 19, 24, and 41 eV, respectively.
  • the terminal elimination half-life was 1.5 and 0.4 hours, respectively, and the injection AUC INF was 4644h*nM and 6511h*nM, respectively.
  • the terminal elimination half-life of the two was 3.0 and 0.7 hours, the peak time was 0.5 and 0.4 hours, the peak concentration was 4622 and 15946nM, respectively, and the oral AUC INF was 12994h*nM and 31406h*nM, respectively.
  • the rats absorbed well, and the oral bioavailability was 28% and 48%, respectively, which was significantly better than the oral bioavailability of the positive reference PLN-74809 (the actual measured data of rats was only 2%).
  • Other compounds in the present invention also have similar physical and chemical properties, and have excellent drug-forming potential and excellent in vivo metabolic characteristics.
  • the experimental test indicators include animal weight and histopathological score.
  • the animal weight is observed and recorded once a day, and the weight growth rate is calculated.
  • the histopathological scoring criteria are as follows:
  • Example 71 helps to improve the overall state of the animals, helps rats recover their weight faster, and no obvious side effects are observed.
  • Example 71 can reduce bleomycin-induced bronchial and arteriolar damage and inflammatory infiltration in the left lung tissue.
  • Example 71 H&E stained sections were observed under a microscope to compare the lesions around the left lung bronchi, pulmonary small blood vessels, and alveolar tissue and the degree of inflammatory cell infiltration. As shown in Figure 4, the low-dose group (D) and high-dose group (E) of Example 71 were significantly different from the model group.
  • the low-dose group (D) and high-dose group (E) of Example 71 can reduce the degree of inflammation of the lung tissue at the edge of the lesion, which is significantly better than the model group (B), and the high-dose group (E) of Example 71 is better than the low-dose group and the positive reference nintedanib group (C) in reducing inflammation.
  • the alveolar wall thickening in the high-dose group (E) of Example 71 was the mildest, with slight alveolar wall thickening at the arrow point, and the degree of pathological changes was significantly lower than that in the model group (B), and also lower than that in the low-dose group (D) and the nintedanib group (C) of Example 71.
  • the degree of reduction in the low-dose group of Example 71 was comparable to that in the nintedanib group. This indicates that the high-dose group of Example 71 is more advantageous than the positive reference nintedanib group, and the drug effect is more significant.
  • Example 71 can alleviate bleomycin-induced pulmonary fibrosis changes
  • the deposition degree of fibrotic collagen in lung tissue was observed by Masson staining under a microscope, and fibrosis scores were performed. As shown in Figure 9, the degree of fibrosis in the low-dose group (D) of Example 71 and the high-dose group (E) of Example 71 was significantly reduced compared with the model group (B), and the degree of reduction in the two dosage groups of Example 71 was better than that in the nintedanib group (C).
  • Figure 11 is the result of statistical analysis of the proportion of mild and severe fibrosis in the sham operation group, model group, nintedanib group, low-dose group of Example 71, and high-dose group of Example 71.
  • fibrosis scores ⁇ 3 points can be regarded as mild; scores ⁇ 4 points can be regarded as severe.
  • the proportion of severe cases in the low-dose group of Example 71 was significantly reduced (p ⁇ 0.001), and the proportion of severe cases in the high-dose group of Example 71 was significantly reduced (p ⁇ 0.001).
  • Example 71 Compared with the positive reference nintedanib group, the proportion of severe cases in the high and low dose groups of Example 71 was significantly lower than that in the nintedanib group, and the proportion of severe cases in the high-dose group of Example 71 was lower than that in the low-dose group, and significantly lower than that in the nintedanib group. This shows that more than 80% of the rats in the high-dose group of Example 71 were mild cases, and the efficacy was extremely significant. Figures 9, 10 and 11 illustrate that Example 71 has a significant anti-pulmonary fibrosis effect.
  • FIG12 shows the degree of collagen deposition in the alveolar tissue of rats in each group in Masson staining.
  • the collagen deposition in the low-dose group (D) of Example 71 and the high-dose group (E) of Example 71 was the lightest, and the degree was significantly lower than that in the model group (B) and the nivolumab group (E). Nintedanib group (C).
  • the collagen deposition area of the high and low dose groups of Example 71 was significantly reduced (both p ⁇ 0.001), and the degree of reduction was significantly better than that of the nintedanib group (p ⁇ 0.05).
  • the degree of reduction in collagen deposition area in the high dose group of Example 71 was more significant, indicating that Example 71 can also effectively reduce the degree of lung collagen deposition.
  • mice (C57BL/6J) were randomly divided into groups according to body weight. From day 2, the normal feed in all cages except the normal control group was replaced with feed containing 0.1% DDC (3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine) to induce mouse sclerosing cholangitis (PSC).
  • the normal control group was provided with normal diet and drinking water for 3 consecutive weeks (Day 2-Day 22). From day 2, the corresponding drugs were given to different groups every day for 3 consecutive weeks (Day 2-Day 22). The day after the last administration (Day 23), the mice were bled, and the blood samples were centrifuged at 4°C and 1500 ⁇ g to separate the serum and frozen at -80°C.
  • mice After the blood was drawn, the mice were euthanized, the whole liver was separated and weighed, and then 1/2 of the liver lobe was cut and placed in 10% formalin (formaldehyde) solution for fixation for 48h, and then pathological examination was performed.
  • formalin formalin
  • the experimental test indicators included animal body weight, liver weight, serum liver function indexes, and histopathological scores. The animal body weight was observed and recorded once a day, and the percentage of body weight change was compared. An automatic biochemical detector was used to measure the levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), total bile acid (TBA), and total bilirubin (TBIL) in mouse blood samples.
  • ALT alanine aminotransferase
  • ALP alkaline phosphatase
  • AST aspartate aminotransferase
  • TAA total bile acid
  • TBIL total bilirubin
  • Example 71 After mice ingested DDC, liver damage gradually worsened, and obvious symptoms of bile duct sclerosis appeared. The body weight of all mice fed with DDC-containing feed decreased to varying degrees, and became more serious over time. The liver weight/body weight ratio increased significantly in the model group, and Example 71 had a significant therapeutic effect compared with the model group, indicating that Example 71 has excellent anti-liver fibrosis, anti-bile duct sclerosis and anti-inflammatory effects.

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Abstract

L'invention concerne un composé inhibiteur de l'intégrine αvβ1, αvβ6 et αvβ8 bicyclique représenté par la formule (I), et un racémate, un stéréoisomère, un tautomère, un solvate, un polymorphe, un métabolite, un ester, un promédicament ou un sel pharmaceutiquement acceptable de celui-ci, et une composition pharmaceutique le comprenant, son procédé de préparation et son utilisation pharmaceutique. La structure de formule (I) est présentée ci-dessous.
PCT/CN2023/124799 2022-10-17 2023-10-16 Dérivé bicyclique utilisé en tant qu'inhibiteur de l'intégrine WO2024083086A1 (fr)

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005007656A1 (fr) * 2003-07-18 2005-01-27 Virochem Pharma Inc. Composes spiro et procedes pour moduler une activite de recepteur de chimiokime
WO2008012623A1 (fr) * 2006-07-25 2008-01-31 Pfizer Products Inc. Composés de benzimidazolyle constituant des potentialisateurs du sous-type de récepteur de glutamate mglur2
CN101534822A (zh) * 2006-09-15 2009-09-16 先灵公司 治疗疼痛、糖尿病和脂质代谢紊乱
WO2010129729A1 (fr) * 2009-05-07 2010-11-11 Merck Sharp & Dohme Corp. Amines spirocycliques substituées utiles en tant que composés antidiabétiques
WO2015200674A1 (fr) * 2014-06-25 2015-12-30 The General Hospital Corporation Composés neuroactifs et leurs procédés d'utilisation
CN108884096A (zh) * 2016-02-08 2018-11-23 豪夫迈·罗氏有限公司 作为ddr1抑制剂的螺二氢吲哚酮
CN109996541A (zh) * 2016-09-07 2019-07-09 普利安特治疗公司 N-酰基氨基酸化合物及其使用方法

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005007656A1 (fr) * 2003-07-18 2005-01-27 Virochem Pharma Inc. Composes spiro et procedes pour moduler une activite de recepteur de chimiokime
WO2008012623A1 (fr) * 2006-07-25 2008-01-31 Pfizer Products Inc. Composés de benzimidazolyle constituant des potentialisateurs du sous-type de récepteur de glutamate mglur2
CN101534822A (zh) * 2006-09-15 2009-09-16 先灵公司 治疗疼痛、糖尿病和脂质代谢紊乱
WO2010129729A1 (fr) * 2009-05-07 2010-11-11 Merck Sharp & Dohme Corp. Amines spirocycliques substituées utiles en tant que composés antidiabétiques
WO2015200674A1 (fr) * 2014-06-25 2015-12-30 The General Hospital Corporation Composés neuroactifs et leurs procédés d'utilisation
CN108884096A (zh) * 2016-02-08 2018-11-23 豪夫迈·罗氏有限公司 作为ddr1抑制剂的螺二氢吲哚酮
CN109996541A (zh) * 2016-09-07 2019-07-09 普利安特治疗公司 N-酰基氨基酸化合物及其使用方法

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