CN111788193B - PD-1/PD-L1 small molecule inhibitor and application thereof in medicines - Google Patents
PD-1/PD-L1 small molecule inhibitor and application thereof in medicines Download PDFInfo
- Publication number
- CN111788193B CN111788193B CN201980016053.3A CN201980016053A CN111788193B CN 111788193 B CN111788193 B CN 111788193B CN 201980016053 A CN201980016053 A CN 201980016053A CN 111788193 B CN111788193 B CN 111788193B
- Authority
- CN
- China
- Prior art keywords
- methoxy
- cancer
- chloro
- biphenyl
- dimethyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- 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
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D207/12—Oxygen or sulfur atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/62—Oxygen or sulfur atoms
- C07D213/70—Sulfur atoms
- C07D213/71—Sulfur atoms to which a second hetero atom is attached
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D213/84—Nitriles
- C07D213/85—Nitriles in position 3
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D455/00—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
- C07D455/02—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing not further condensed quinolizine ring systems
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
A PD-1/PD-L1 small molecule inhibitor and application in medicine thereof, in particular to a PD-1/PD-L1 small molecule inhibitor which is a compound shown as a formula (I) or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, an ester, a pharmaceutically acceptable salt or a prodrug of the compound shown as the formula (I); also provides a preparation method of the compound shown in the formula (I), a pharmaceutical composition and application thereof in preparing medicaments for preventing or treating diseases related to PD-1/PD-L1 signal pathways.
Description
Cross-reference to related applications:
this application claims priority to the citation of a chinese patent application No. 201810203812.4, earlier filed on the chinese intellectual property office at application date 2018.03.13, the entire contents of which are incorporated herein by reference.
Technical Field
The invention discloses a small molecular compound used as an inhibitor of PD-1/PD-L1 protein interaction, a preparation method thereof, a pharmaceutical composition and application, and belongs to the field of medicines.
Background
With the continuous development of social economy, the gradual improvement of the living standard of people, the change of the dietary structure of residents, the aging of population, urbanization, continuous deterioration of environment and other factors, the disease spectrum and the death spectrum of China are obviously changed, and the incidence of malignant tumors is on a continuously rising trend. Malignant tumors have become the first killers which are harmful to the life health of human beings and restrict the development of social economy. With the improvement of medical technology level, although the methods for resisting tumors of human beings are continuously innovated, most of tumor patients are relieved and cured by the current clinically applied operations, chemotherapies, radiotherapies and newly appeared molecular targeted drug treatments, but part of the patients still have poor sensitivity to the treatment means and poor prognosis. Immunotherapy offers a long-lasting therapeutic effect compared to other types of tumor therapy, and has gradually shifted from initial non-specific immunotherapy to specific immune-targeted therapy. In recent decades, the milestone success of immune checkpoint inhibitors and chimeric antigen receptor T cell therapies has pushed tumor immunity to become an effective cancer treatment means following surgery, radiotherapy, chemotherapy and targeted therapy.
The mechanism of immunotherapy is to activate specific T cells, target attack to eliminate tumor cells, and activate anti-tumor immunity in patientsAnd (5) the system responds. T cell activation is primarily referred to as "dual signal" intervention, i.e., the first signal is provided by the antigenic peptide-major histocompatibility complex and the T cell antigen receptor complex, CD 28 Costimulatory molecules such as B7 provide second messengers for T cell activation. When the stimulation of the synergistic stimulation molecules is lacked, the imbalance of T cell response is easily caused, so that the tumor escapes from the organism for immune monitoring. As a second signal for activating T cells, activation of a programmed death receptor 1 (PD-1) and a corresponding ligand (programmed death-ligand 1, PD-L1) signal pathway can inhibit an organism anti-tumor immune response, so that the pathway is blocked by a medicament to obviously inhibit the growth of tumors.
PD-1 is also named CD279, is an immune co-suppression molecule found in apoptotic T cell hybridoma by subtractive hybridization technology, is a type I transmembrane protein consisting of 268 amino acids, and belongs to CD 28 A member of a family. PD-1 is structurally composed mainly of an extracellular Ig V-like domain, a hydrophobic transmembrane region, and a cytoplasmic region 3 that retains an Immunoreceptor Tyrosine Inhibition Motif (ITIM) and an immunoreceptor tyrosine transformation motif (ITSM). PD-1 is expressed not only in immune cells such as activated T lymphocytes, B lymphocytes, natural Killer (NK) cells and monocytes, but also on some tumor cell lines or on the surface of tumor cells. PD-L1 is a natural ligand of PD-1, and in a physiological condition, a negative signal generated by the interaction of PD-1 and PD-L1 is a 'braking' mechanism, so that additional injury caused by excessive immune response is mainly prevented. For example, PD-1 of peripheral autoreactive T cells interacts with PD-L1 on the surface of non-hematopoietic cells in tissues, and can suppress proliferation and cytokine secretion of the T cell population, thereby achieving an immune tolerance effect. The tumor immune process mainly occurs in the periphery, and the T cells are excited and matured by antigen presenting cells in the primary lymphatic system to show high expression of PD-1. Tumor cells are stimulated by extracellular high levels of cytokines or by sustained activation of certain pathways, which up-regulate PD-L1 expression. Once tumors are infiltrated by peripheral T cells, excessive negative regulation signals are generated by the combination of a large amount of PD-1 and PD-L1, so that the original active T Cell Receptor (TCR) downstreamThe signal is inhibited, the T cell gradually loses energy and even dies, and the tumor realizes immune escape.
Monoclonal antibody drugs targeting PD-1 were introduced in 2014 as Pembrolizumab (Merck) and Nivolumab (Nivolumab) of Bristol-Myers Squibb (BMS), respectively, for the treatment of metastatic melanoma. PD-L1 mAb Abterozumab (Atezolizumab), developed by the Lowen Sulqin Gene Take (Genentech), was approved by the FDA in the United states for the treatment of bladder cancer at month 3 of 2016 and was approved for the treatment of lung cancer at month 11 of the same year. Unlike the well-established fire in the antibody drug market, the development of small molecule immune checkpoint blockers has not presented a pattern of blowout. The small molecule inhibitors reported in the prior art comprise benzene sulfonamide compounds in Harvarsharpe topic group, 2,6-disubstituted toluene immunomodulator disclosed by Baishimei noble and peptidomimetic molecules developed by Curis/Aurigene cooperation, wherein the IND application of the United states FDA is obtained in 2016 and 6 months only from the small peptidomimetic molecules developed by Curis/Aurigene cooperation, and is currently in clinical stage.
Compared with antibody drugs, the small-molecule inhibitor has controllable pharmacokinetic behaviors, is relatively low in cost, and well makes up for clinical defects of large-molecule drugs. Therefore, the design and synthesis of the small molecule inhibitor with the blocking effect aiming at the PD-1/PD-L1 have practical significance.
Summary of The Invention
The invention provides a small molecule inhibitor for inhibiting PD-1/PD-L1 interaction, which is a compound shown in formula (I) or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, an ester, a pharmaceutically acceptable salt or a prodrug of the compound shown in formula (I), and provides a preparation method of the compound shown in formula (I), a pharmaceutical composition and application thereof in preparing medicines for preventing or treating diseases related to a PD-1/PD-L1 signal pathway.
Detailed Description
In order to solve the technical problem, the invention provides the following technical scheme:
the first aspect of the technical scheme of the invention provides a compound of a PD-1/PD-L1 small molecule inhibitor, which is a structure shown in a formula (I) or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, an ester, a pharmaceutically acceptable salt or a prodrug of the structure shown in the formula (I),
in the formula (I), the compound is shown in the specification,
R 1 is- (CH) 2 ) n Ar, wherein Ar is C 6-12 Aryl or C 5-12 A heteroaryl group; ar is optionally substituted with 1,2,3 or 4 substituents each independently being H, D, cyano, halogen, amino, methanesulfonyl, acetylamino, C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or C 1-6 An alkoxy group; n is 1,2,3 or 4;
a is-CH 2 O-、-OCH 2 -、-CH 2 -CH 2 -, -C (O) NH-or-NHC (O) -;
R 2 is composed of
Wherein R is 5 And R 6 Each independently of the other being H, halogen, C 1-6 Alkyl radical, C 1-6 Haloalkyl or C 1-6 An alkoxy group; z is CH or N;
R 7 is composed of
q is 1,2,3 or 4;
R v is composed of
WhereinSaid R is v May be optionally substituted with 0,1, 2 or 3 substituents selected from hydroxy, cyano, amino, oxo (= O), C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy or amino C 1-6 Alkyl radical, W 1 、W 3 And W 5 Each independently is CH 2 、S、O、S(O) 2 Or NH, W 2 And W 4 Each independently is CH or N; m1, m2, m3, m4, m5, m6, m7 and m8 are each independently 0,1, 2 or 3;
R 3 is C 1-8 Alkylamino or C 3-9 Heterocyclic group, wherein, said C 3-9 The heterocyclic group contains at least one N atom; said C 1-8 Alkylamino or C 3-9 The heterocyclyl is optionally substituted with 0,1, 2,3 or 4 substituents selected from hydrogen, hydroxy, halogen, carboxy, C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy, amino C 1-6 Alkyl, acetylamino, cyano, sulfonamido or oxo (= O);
p is 0,1, 2 or 3;
x and Y are each independently H, D, C 1-6 Alkyl radical, C 1-6 Alkoxy, halogen or cyano.
In some embodiments, ar described herein is phenyl, pyridyl, pyrimidinyl, indolyl, or quinolinyl; ar is optionally substituted with 1,2,3 or 4 substituents each independently being H, D, cyano, halogen, amino, methanesulfonyl, acetylamino, C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or C 1-6 An alkoxy group.
In some embodiments, R as described herein v Is one of the groups formed by the following structural formula:
the W is 1 、W 3 And W 5 Each independently CH2, S, O, S (O) 2 Or NH, W 2 And W 4 Each independently is CH or N; r is v Optionally substituted with 0,1, 2 or 3 substituents selected from hydroxy, cyano, amino, oxo (= O), C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy or amino C 1-6 An alkyl group.
In other embodiments, R is as described herein v Is one of the groups formed by the following structural formula:
wherein, R is v Optionally substituted with 0,1, 2 or 3 substituents selected from hydroxy, cyano, amino, oxo (= O), C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy or amino C 1-6 An alkyl group.
In some embodiments, R is as described herein 5 And R 6 Each independently H, D, fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, tert-butyl, isobutyl, n-butyl, trifluoromethyl, difluoromethyl, monofluoromethyl, methoxy, or ethoxy.
In some embodiments, R is as described herein 3 Is one of the groups formed by the following structures:
in some embodiments, the invention provides a small molecule inhibitor for inhibiting PD-1/PD-L1 interaction, which is a compound shown as formula (II), or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, an ester, a pharmaceutically acceptable salt or a prodrug thereof,
in the formula (I), the compound is shown in the specification,
R 1 is- (CH) 2 ) n Ar, wherein Ar is phenyl, pyridyl, pyrimidyl, indolyl or quinolyl; ar is optionally substituted with 1,2,3 or 4 substituents each independently being H, cyano, halogen, amino, methanesulfonyl, acetylamino, C 1-6 Alkyl or C 1-6 An alkoxy group; n is 1,2,3 or 4;
R 2 、R 3 、R 5 、R 6 、R v x, Y, Z and p have the meaning described in the present invention.
In one aspect, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) or formula (II) of the present invention, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt, or a prodrug thereof, and a pharmaceutically acceptable adjuvant, or a combination thereof.
In one aspect, the invention relates to the use of a compound of formula (I) or (II) or a pharmaceutical composition thereof for the manufacture of a medicament for the prevention, treatment or alleviation of a disease or a disorder related to the PD-1/PD-L1 signalling pathway in a patient.
In some embodiments, the disease associated with the PD-1/PD-L1 signaling pathway described herein is cancer, an infectious disease, or an autoimmune disease.
In some embodiments, the cancer of the invention is a disease in which there is unlimited proliferation of cells in an organ or a body tissue; the infectious diseases are bacterial infectious diseases, viral infectious diseases or fungal infectious diseases; the autoimmune disease is an organ-specific autoimmune disease or a systemic autoimmune disease.
In some embodiments, the disease of unlimited proliferation of cells in an organ or body tissue described herein is bone cancer, head and neck cancer, pancreatic cancer, skin cancer, malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, hodgkin's disease, non-hodgkin's lymphoma, carcinoma of the esophagus, carcinoma of the small intestine, cancer of the endocrine system, carcinoma of the thyroid gland, carcinoma of the parathyroid gland, carcinoma of the adrenal gland, sarcoma of soft tissue, carcinoma of the urethra, carcinoma of the penis, chronic or acute leukemia, solid tumor of the child, lymphocytic lymphomas, carcinoma of the bladder, carcinoma of the kidney or ureter, carcinoma of the renal pelvis, tumor of the Central Nervous System (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, or an environmentally induced cancer or a combination of the foregoing; wherein the chronic or acute leukemia comprises acute myelocytic leukemia, chronic myelocytic leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia.
In some embodiments, the viral infectious disease described herein is aids, hepatitis a, hepatitis b, hepatitis c, hepatitis d, herpes virus infection, papillomavirus infection, and influenza.
In some embodiments, the organ-specific immune disorder of the invention is chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis of the cerebrospinal fluid and acute idiopathic polyneuritis; the systemic autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease and autoimmune hemolytic anemia. In another aspect, the invention provides a method of modulating an immune response in a subject mediated by the PD-1 signaling pathway, comprising administering to the subject a therapeutically effective amount of a compound of the invention, thereby modulating the immune response in the subject.
In some embodiments, the invention features a method of protecting, managing, treating, or ameliorating a disease associated with the PD-1/PD-L1 signaling pathway in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound or pharmaceutical composition of the invention.
In some embodiments, the disease associated with the PD-1/PD-L1 signaling pathway described herein is cancer, an infectious disease, or an autoimmune disease.
In some embodiments, the cancer of the invention is a disease in which there is unlimited proliferation of cells in an organ or a body tissue; the infectious diseases are bacterial infectious diseases, viral infectious diseases or fungal infectious diseases; the autoimmune disease is an organ-specific autoimmune disease or a systemic autoimmune disease.
In some embodiments, the disease of unlimited proliferation of cells in an organ or body tissue described herein is bone cancer, head and neck cancer, pancreatic cancer, skin cancer, malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, hodgkin's disease, non-hodgkin's lymphoma, carcinoma of the esophagus, carcinoma of the small intestine, cancer of the endocrine system, carcinoma of the thyroid gland, carcinoma of the parathyroid gland, carcinoma of the adrenal gland, sarcoma of soft tissue, carcinoma of the urethra, carcinoma of the penis, chronic or acute leukemia, solid tumor of the child, lymphocytic lymphomas, carcinoma of the bladder, carcinoma of the kidney or ureter, carcinoma of the renal pelvis, tumor of the Central Nervous System (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, or an environmentally induced cancer or a combination of the foregoing; wherein the chronic or acute leukemia comprises acute myelocytic leukemia, chronic myelocytic leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia.
In some embodiments, the viral infectious disease described herein is aids, hepatitis a, hepatitis b, hepatitis c, hepatitis d, herpes virus infection, papillomavirus infection, and influenza.
In some embodiments, the organ-specific immune disorder of the invention is chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis of the cerebrospinal fluid and acute idiopathic polyneuritis; the systemic autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease and autoimmune hemolytic anemia. In another aspect, the invention provides a method of modulating an immune response in a subject mediated by the PD-1 signaling pathway, comprising administering to the subject a therapeutically effective amount of a compound of the invention, thereby modulating the immune response in the subject.
In some embodiments, the compounds or pharmaceutical compositions described herein are used to protect, manage, treat, or ameliorate a disease associated with the PD-1/PD-L1 signaling pathway in a patient.
In some embodiments, the disease associated with the PD-1/PD-L1 signaling pathway described herein is cancer, an infectious disease, or an autoimmune disease.
In some embodiments, the cancer of the invention is a disease in which there is unlimited proliferation of cells in an organ or a body tissue; the infectious diseases are bacterial infectious diseases, virus infectious diseases or fungal infectious diseases; the autoimmune disease is an organ-specific autoimmune disease or a systemic autoimmune disease.
In some embodiments, the disease of unlimited proliferation of cells in an organ or body tissue described herein is bone cancer, head and neck cancer, pancreatic cancer, skin cancer, malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, hodgkin's disease, non-hodgkin's lymphoma, carcinoma of the esophagus, carcinoma of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemia, solid tumor of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, central Nervous System (CNS) tumor, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, or an environmentally-induced cancer, or a combination of the foregoing; wherein the chronic or acute leukemia comprises acute myelocytic leukemia, chronic myelocytic leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia.
In some embodiments, the viral infectious disease of the present invention is aids, hepatitis a, hepatitis b, hepatitis c, hepatitis d, herpes virus infection, papillomavirus infection, and influenza.
In some embodiments, the organ-specific immune disease of the invention is chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis, and acute idiopathic polyneuritis; the systemic autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease and autoimmune hemolytic anemia.
In another aspect, the invention provides a method of modulating an immune response in a subject mediated by the PD-1 signaling pathway, comprising administering to the subject a therapeutically effective amount of a compound of the invention, thereby modulating the immune response in the subject.
In another aspect, the invention relates to a process for the preparation, isolation and purification of the compounds encompassed by formula (I) or (II).
The foregoing merely summarizes certain aspects of the invention and is not intended to be limiting. Other aspects will be more fully described in detail below.
Detailed description of the invention
Definitions and general terms
Reference will now be made in detail to certain embodiments of the invention, examples of which are illustrated by the accompanying structural and chemical formulas. The invention is intended to cover alternatives, modifications and equivalents, which may be included within the scope of the invention as defined by the appended claims. Those skilled in the art will recognize that many methods and materials similar or equivalent to those described herein can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described herein. In the event that one or more of the incorporated documents, patents, and similar materials differ or contradict this application (including but not limited to defined terminology, application of terminology, described techniques, and the like), this application controls.
It will be further appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications referred to herein are incorporated by reference in their entirety.
The following definitions as used herein should be applied unless otherwise indicated. For the purposes of the present invention, the chemical elements are in accordance with the CAS version of the periodic Table of the elements, and the handbook of chemistry and Physics, 75 th edition, 1994. In addition, reference may be made to the general principles of Organic Chemistry, thomas Sorrell, university Science Books, sausaltito: 1999, and "March's Advanced Organic Chemistry" by Michael B. Smith and Jerry March, john Wiley & Sons, new York:2007, the entire contents of which are incorporated herein by reference.
The articles "a," "an," and "the" as used herein are intended to include "at least one" or "one or more" unless otherwise indicated or clearly contradicted by context. Thus, as used herein, the articles refer to articles of one or more than one (i.e., at least one) object. For example, "a component" refers to one or more components, i.e., there may be more than one component contemplated to be employed or used in embodiments of the described embodiments.
The term "subject" as used herein refers to an animal. Typically the animal is a mammal. Subjects, e.g., also primates (e.g., humans, males or females), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds, etc. In certain embodiments, the subject is a primate. In other embodiments, the subject is a human.
The term "patient" as used herein refers to humans (including adults and children) or other animals. In some embodiments, "patient" refers to a human.
The term "comprising" is open-ended, i.e. includes the elements indicated in the present invention, but does not exclude other elements.
"stereoisomers" refers to compounds having the same chemical structure, but differing in the arrangement of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric isomers (cis/trans), atropisomers, and the like.
"chiral" is a molecule having the property of not overlapping its mirror image; and "achiral" refers to a molecule that can overlap with its mirror image.
"enantiomer" refers to two isomers of a compound that are not overlapping but are in mirror image relationship to each other.
"diastereomer" refers to a stereoisomer having two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, and reactivities. Mixtures of diastereomers may be separated by high resolution analytical procedures such as electrophoresis and chromatography, e.g., HPLC.
The stereochemical definitions and rules used in the present invention generally follow S.P. Parker, ed., mcGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, new York; and Eliel, E.and Wilen, S., "Stereochemistry of Organic Compounds", john Wiley & Sons, inc., new York,1994.
Many organic compounds exist in an optically active form, i.e., they have the ability to rotate the plane of plane polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of a molecule with respect to one or more of its chiral centers. The prefixes d and l or (+) and (-) are the symbols used to specify the rotation of plane polarized light by the compound, where (-) or l indicates that the compound is left-handed. Compounds prefixed with (+) or d are dextrorotatory. A particular stereoisomer is an enantiomer and a mixture of such isomers is referred to as an enantiomeric mixture. A50: 50 mixture of enantiomers is referred to as a racemic mixture or racemate, which may occur when there is no stereoselectivity or stereospecificity in the chemical reaction or process.
Any asymmetric atom (e.g., carbon, etc.) of a compound disclosed herein can exist in racemic or enantiomerically enriched forms, e.g., the (R) -, (S) -or (R, S) -configuration. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R) -or (S) -configuration.
Depending on the choice of starting materials and methods, the compounds of the invention may exist as one of the possible isomers or as mixtures thereof, for example as racemates and diastereomeric mixtures (depending on the number of asymmetric carbon atoms). Optically active (R) -or (S) -isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituent may be in the E or Z configuration; if the compound contains a disubstituted cycloalkyl group, the substituents of the cycloalkyl group may have cis or trans configuration.
Any resulting mixture of stereoisomers may be separated into pure or substantially pure geometric isomers, enantiomers, diastereomers, depending on differences in the physicochemical properties of the components, for example, by chromatography and/or fractional crystallization.
The racemates of any of the resulting end products or intermediates can be resolved into the optical enantiomers by known methods using methods familiar to those skilled in the art, e.g., by separation of the diastereomeric salts obtained. The racemic product can also be separated by chiral chromatography, e.g., high Performance Liquid Chromatography (HPLC) using a chiral adsorbent. In particular, enantiomers can be prepared by asymmetric synthesis, for example, see Jacques, et al, enantiomers, racemes and solutions (Wiley Interscience, new York, 1981); principles of Asymmetric Synthesis (2) nd Ed.Robert E.Gawley，Jeffrey Aubé，Elsevier，Oxford，UK，2012)；Eliel，E.L.Stereochemistry of Carbon Compounds(McGraw-Hill，NY，1962)；Wilen，S.H.Tables of Resolving Agents and Optical Resolutions p.268(E.L.Eliel，Ed.，Univ.of Notre Dame Press，Notre Dame，IN 1972)；Chiral Separation Techniques：A Practical Approach(Subramanian，G.Ed.，Wiley-VCH Verlag GmbH&Co.KGaA，Weinheim，Germany，2007)。
The term "tautomer" or "tautomeric form" refers to structural isomers having different energies that can interconvert by a low energy barrier (low energy barrier). If tautomerism is possible (e.g., in solution), then the chemical equilibrium of the tautomer can be reached. For example, proton tautomers (also known as proton transfer tautomers) include interconversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers (valenctautomers) include interconversion by recombination of some of the bonding electrons. A specific example of keto-enol tautomerism is the tautomerism of the pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another example of tautomerism is phenol-ketone tautomerism. One specific example of phenol-ketone tautomerism is the tautomerism of pyridin-4-ol and pyridin-4 (1H) -one tautomers. Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention.
The terms "optional" or "optionally" mean that the subsequently described event or circumstance may, but need not, occur, i.e., that the description includes instances where said event or circumstance occurs and instances where it does not. For example, "optionally substituted with 1,2,3, or 4.. Includes the case where the group is substituted with 1, or 2, or 3, or 4 of the substituents described, and the case where the group is not substituted with the substituents described. Further, when the group is substituted with 1 or more of the substituents described above, the substituents are independent of each other, that is, the 1 or more substituents may be different from each other or the same.
The compounds of the invention may be independently optionally substituted with one or more substituents, as described herein, in the general formula above, or as specified in the examples, subclasses, and classes of compounds encompassed by the invention. It is understood that the term "optionally substituted" may be used interchangeably with the term "substituted or unsubstituted". The term "independently optionally" is used interchangeably with the term "optionally independently", and generally speaking, the term "independently optionally" whether or not preceded by the term "substituted" indicates that one or more hydrogen atoms in a given structure may be substituted or unsubstituted with the particular substituent 1. Unless otherwise indicated, an optional substituent group may have a substituent 1 substituted at each substitutable position of the group. When more than one position in a given formula can be substituted with one or more substituents 1 selected from a particular group, the substituents may be substituted at each position, identically or differently. Wherein said substituent 1 may be, but is not limited to: hydrogen, oxo (= O), fluoro, chloro, bromoIodine, hydroxy, amino, carboxy, alkyl-S (= O) t -, haloalkyl, hydroxyalkyl, aminoalkyl, aldehyde group, aminoacyl, alkoxy, alkylamino, alkylthio, haloalkoxy, cyano, aryl, heteroaryl, alkenyl, alkynyl, heterocyclic group, mercapto, nitro, aryloxy, hydroxyalkoxy, alkyl- (C = O) -, benzyl, cyclopropyl, phenyl, alkylamino-C (= O) -, CN-alkyl-C (= O) -, alkyl-O-C (= O) -alkyl, ester group, hydroxyalkylacyl, or alkoxyalkyl, and the like. Where appropriate, the substituents 1 can furthermore be optionally mono-substituted independently or polysubstituted identically or differently by substituents 2. Wherein said substituent 2 may be, but is not limited to: hydrogen, oxo (= O), fluoro, chloro, bromo, iodo, hydroxy, amino, carboxy, alkyl-S (= O) t -, haloalkyl, hydroxyalkyl, aminoalkyl, aldehyde group, aminoacyl, alkoxy, alkylamino, alkylthio, haloalkoxy, cyano, aryl, heteroaryl, alkenyl, alkynyl, heterocyclic group, mercapto, nitro, aryloxy, hydroxyalkoxy, alkyl- (C = O) -, benzyl, cyclopropyl, phenyl, alkylamino-C (= O) -, CN-alkyl-C (= O) -, alkyl-O-C (= O) -alkyl, ester group, hydroxyalkylacyl, or alkoxyalkyl, and the like. Wherein t is 0,1, or 2.
In the various parts of this specification, substituents of the disclosed compounds are disclosed in terms of group type or range. It is specifically intended that the invention includes each and every independent subcombination of the various members of these groups and ranges. For example, the term "C 1-6 Alkyl "means in particular independently disclosed methyl, ethyl, C 3 Alkyl radical, C 4 Alkyl radical, C 5 Alkyl and C 6 An alkyl group.
In each of the parts of the invention, linking substituents are described. Where the structure clearly requires a linking group, the markush variables listed for that group are understood to be linking groups. For example, if the structure requires a linking group and the markush group definition for the variable recites "alkyl" or "aryl," it is understood that the "alkyl" or "aryl" represents an attached alkylene group or arylene group, respectively.
The term "alkyl" as used herein includes saturated straight or branched chain monovalent hydrocarbon groups of 1 to 20 carbon atoms, wherein the alkyl groups may independently be optionally substituted with one or more substituents described herein. In some of these embodiments, the alkyl group contains 1 to 10 carbon atoms; in still other embodiments, the alkyl group contains 1 to 8 carbon atoms; in other embodiments, the alkyl group contains 1 to 6 carbon atoms, and in other embodiments, the alkyl group contains 1 to 4 carbon atoms; in other embodiments, the alkyl group contains 1 to 3 carbon atoms. Still further examples of alkyl groups include, but are not limited to, methyl (Me, -CH) 3 ) Ethyl (Et, -CH) 2 CH 3 ) N-propyl (n-Pr, -CH) 2 CH 2 CH 3 ) Isopropyl (i-Pr, -CH (CH) 3 ) 2 ) N-butyl (n-Bu, -CH) 2 CH 2 CH 2 CH 3 ) 2-methylpropyl or isobutyl (i-Bu, -CH) 2 CH(CH 3 ) 2 ) 1-methylpropyl or sec-butyl (s-Bu, -CH (CH) 3 )CH 2 CH 3 ) Tert-butyl (t-Bu, -C (CH) 3 ) 3 ) N-pentyl (-CH) 2 CH 2 CH 2 CH 2 CH 3 ) 2-pentyl (-CH (CH) 3 )CH 2 CH 2 CH 3 ) 3-pentyl (-CH (CH) 2 CH 3 ) 2 ) 2-methyl-2-butyl (-C (CH) 3 ) 2 CH 2 CH 3 ) 3-methyl-2-butyl (-CH (CH) 3 )CH(CH 3 ) 2 ) 3-methyl-1-butyl (-CH) 2 CH 2 CH(CH 3 ) 2 ) 2-methyl-1-butyl (-CH) 2 CH(CH 3 )CH 2 CH 3 ) N-hexyl (-CH) 2 CH 2 CH 2 CH 2 CH 2 CH 3 ) 2-hexyl (-CH (CH) 3 )CH 2 CH 2 CH 2 CH 3 ) 3-hexyl (-CH (CH) 2 CH 3 )(CH 2 CH 2 CH 3 ) 2-methyl-2-pentyl (-C (CH)) 3 ) 2 CH 2 CH 2 CH 3 ) 3-methyl-2-pentyl (-CH (CH) 3 )CH(CH 3 )CH 2 CH 3 ) 4-methyl-2-pentyl (-CH (CH) 3 )CH 2 CH(CH 3 ) 2 ) 3-methyl-3-pentyl (-C (CH) 3 )(CH 2 CH 3 ) 2 ) 2-methyl-3-pentyl (-CH (CH) 2 CH 3 )CH(CH 3 ) 2 ) 2,3-dimethyl-2-butyl (-C (CH) 3 ) 2 CH(CH 3 ) 2 ) 3,3-dimethyl-2-butyl (-CH (CH) 3 )C(CH 3 ) 3 ) N-heptyl, n-octyl, and the like. The term "alkyl" and its prefix "alkane" as used herein, both include straight and branched saturated carbon chains. Alkyl groups may be substituted with the substituents described herein.
The term "haloalkyl" denotes a situation where an alkyl group may be substituted by one or more of the same or different halogen atoms. Wherein the alkyl group has the meaning as described herein, examples include, but are not limited to, trifluoromethyl, difluoromethyl, monofluoromethyl, 2,2-difluoroethyl, 3,3,3-trifluoropropyl, and the like. Haloalkyl groups may be substituted with substituents described herein.
The term "amino" refers to a compound having the formula-NH 2 A group of (2).
The term "carboxy", whether used alone or in combination with other terms, means-CO 2 H; the term "carbonyl", whether used alone or in combination with other terms, means- (C = O) -.
The term "alkylamino" or "alkylamino" refers to "N-alkylamino" or "N, N-alkylamino" in which the hydrogen atom of an amino group is replaced by one or two separate alkyl groups, wherein the alkyl groups have the meaning as described herein. In some of these embodiments, the alkylamino group is a C 1-8 Lower alkylamino groups in which the alkyl group is attached to the nitrogen atom. In other embodiments, the alkylamino group is C 1-3 Lower alkylamino groups of (a). Examples include, but are not limited to, N-methylamino, N-ethylamino, N, N-dimethylamino, and the like. "alkylamino" or "alkylamino" may be substituted with the substituents described herein. The term "C 1-8 Alkylamino means alkylaminoEach independent alkyl group in the group contains 1 to 8 carbon atoms and the alkylamino group has the meaning as defined above.
The term "alkoxy" means an alkyl group attached to the rest of the molecule through an oxygen atom, wherein the alkyl group has the meaning as described herein. The term "G 1-6 Alkoxy "means an alkyl group having 1 to 6 carbon atoms attached to the rest of the molecule through an oxygen atom. In some embodiments, alkoxy groups contain 1 to 6 carbon atoms; in other embodiments, the alkoxy group contains 1 to 4 carbon atoms; in still other embodiments, alkoxy groups contain 1-3 carbon atoms. The alkoxy group may be optionally substituted with one or more substituents described herein. Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH) 3 ) Ethoxy (EtO, -OCH) 2 CH 3 ) 1-propoxy (n-PrO, n-propoxy, -OCH) 2 CH 2 CH 3 ) And so on.
The term "alkanoyl" or "alkanoyl" means an alkyl group attached to the rest of the molecule through a carbonyl group (-C (= O) -), wherein the alkyl group has the meaning described herein. The alkanoyl group can be optionally substituted with one or more substituents described herein. Examples of alkanoyl groups include, but are not limited to, acetyl (-C (= O) CH 3 ) Propionyl (- = O) CH 2 CH 3 ) Butyryl (-C (= O) CH 2 CH 2 CH 3 ) And so on.
The term "alkanoylamino" or "alkanoylamino" means that an alkanoyl group is attached to the rest of the molecule through a group (-NH-) wherein alkanoyl has the meaning as described herein. The alkanoylamino group may be optionally substituted with one or more substituents described herein. Examples of alkanoylamino groups include, but are not limited to, acetylamino (CH) 3 -C (= O) -NH-), propionylamino (CH) 3 CH 2 -C (= O) -NH-), and the like.
The terms "alkoxyacyl", "alkylaminoacyl", "aminoacyl" denote alkoxy, alkylamino or amino (-NH) 2 ) The group is attached to the rest of the molecule via a carbonyl group (-C (= O) -), wherein alkoxy and alkylamino groups have the meaning as described herein. The alkoxyacyl, alkylaminoacyl, aminoacyl groups may be optionally substituted with one or more substituents described herein. Examples of the alkoxyacyl group, the alkylaminoacyl group include, but are not limited to, methoxyacyl group (- = O) OCH 3 ) Ethoxyacyl (- = O) OCH 2 CH 3 ) Methylaminoacyl (-C (= O) NHCH 3 ) Dimethylaminoyl (- = O) N (CH) 3 ) 2 ) And so on.
The terms "alkylsulfonyl", "alkoxysulfonyl", "alkylaminosulfonyl", "aminosulfonyl" denote alkyl, alkoxy, alkylamino or amino (-NH) 2 ) The radical passing through a sulfonyl group (-SO) 2 -) is attached to the rest of the molecule, wherein the alkyl, alkoxy, alkylamino groups have the meaning as described in the present invention. The alkylsulfonyl, alkoxysulfonyl, alkylaminosulfonyl, aminosulfonyl groups may be optionally substituted with one or more substituents described herein. Examples of alkylsulfonyl groups include, but are not limited to, methylsulfonyl (-SO) 2 CH 3 ) Ethylsulfonyl (-SO) 2 CH 2 CH 3 ) And so on.
The term "aryl" may be monocyclic, bicyclic, and tricyclic carbocyclic ring systems in which at least one ring system is aromatic, wherein each ring system contains 3 to 7 atoms and only one point of attachment is attached to the rest of the molecule. The term "aryl" may be used interchangeably with the term "aromatic ring", e.g., aromatic rings may include phenyl, naphthyl, and anthracene. The aryl group may be substituted with the substituent described in the present invention. The term "C 6-12 Aryl "means an aryl group having 6 to 12 carbon atoms, the aryl group being as defined above.
The terms "heteroaryl", "heteroaryl ring" and "heteroaromatic ring" are used interchangeably herein and all refer to monocyclic, bicyclic, tricyclic or tetracyclic ring systems in which the bicyclic, tricyclic or tetracyclic heteroaromatic ring systems form a ring in fused form. Wherein the heteroaromatic ring system is aromatic as a wholeOptionally, one or more of the ring atoms is independently optionally substituted with a heteroatom (heteroatom selected from N, O, P, S, where S or P is independently optionally substituted with one or more oxygen atoms to give the like SO, SO 2 、PO、PO 2 The group of (1). The heteroaryl system may be attached to the main structure at any heteroatom or carbon atom that results in the formation of a stable compound. The heteroaryl group may be a monocyclic ring of 3 to 7 atoms, or a bicyclic ring of 7 to 10 atoms, or a tricyclic ring of 10 to 15 atoms. The bicyclic ring having 7-10 atoms can be bicyclo [4,5]，[5，5]，[5，6]Or [6,6]A system, the tricyclic ring of 10-15 atoms may be a tricyclic ring [5,5,6]、[5，7，6]Or [6,5,6]And (4) preparing the system. The term "C 5-12 Heteroaryl "means a heteroaryl group having 5 to 12 carbon atoms and the heteroaryl group is as defined above.
In other embodiments, the heteroaryl system (including heteroaryl, heteroaryl ring) includes, but is not limited to, the following examples: furan-2-yl, furan-3-yl, N-imidazolyl, imidazol-2-yl, imidazol-4-yl, imidazol-5-yl, isoxazol-3-yl, isoxazol-4-yl, isoxazol-5-yl, oxazol-2-yl, oxazol-4-yl, oxazol-5-yl, 4-methylisoxazol-5-yl, N-pyrrolyl, pyrrol-2-yl, pyrrol-3-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, pyrimidin-2-yl, pyrimidin-4-yl, pyrimidin-5-yl, pyridazinyl (such as pyridazin-3-yl), thiazol-2-yl, thiazol-4-yl, thiazol-5-yl, tetrazolyl (e.g., tetrazol-5-yl), triazolyl (e.g., triazol-2-yl and triazol-5-yl), thiophen-2-yl, thiophen-3-yl, pyrazolyl (e.g., pyrazol-2-yl), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazol-2-yl, pyrazinyl, pyrazin-2-yl, 1,3,5-triazinyl, benzo [ d ] thiazol-2-yl, imidazo [1,5-a ] pyridin-6-yl, benzimidazolyl, benzoxazolyl, quinoxalinyl, 1,8-naphthyridinyl, benzothienyl, benzothiazolyl, indolyl (e.g., indol-2-yl), purinyl, quinolinyl (e.g., quinolin-2-yl, quinolin-3-yl, quinolin-4-yl), isoquinolinyl (e.g., isoquinolin-1-yl, isoquinolin-3-yl or isoquinolin-4-yl), benzopyrazolyl, acridinyl, benzimidazolyl, benzindolyl, benzisoxazinyl, benzo [4,6] imidazo [1,2-a ] pyridyl, benzo [ d ] imidazo [2,1-b ] thiazolyl, benzothiadiazolyl, benzothiazolyl, benzothiophenyl, benzotriazolyl, benzothiopyranyl, benzoxazinyl, benzoxazolyl, benzothiazolyl, β -carbolinyl, carbazolyl, phthalazinyl, dibenzofuranyl, imidazopyridinyl, imidazothiazolyl, indazolyl, indolizinyl, indolyl, isobenzothianyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, naphthyridinyl, oxazolidinedionyl, oxazolidinyl, oxazolopyridyl, oxazolyl, theadiazenyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, pyridopyridyl, quinazolinyl, quinoxalinyl, thiophenyl, triazinyl, 2H-pyrrolo [3,4-c ] pyridyl, pyrazolo [2',1':2,3] oxazolo [4,5-c ] pyridyl, imidazo [2',1':2,3] thiazolo [4,5-c ] pyridinyl, imidazo [2',1':2,3] thiazolo [4,5-b ] pyridinyl, imidazo [2',1':2,3] thiazolo [5,4-b ] pyridinyl, pyrazolo [2',1':2,3] thiazolo [4,5-b ] pyrazinyl, 1H-benzo [4,5] thieno [2,3-d ] imidazolyl, benzo [4,5] thieno [2,3-d ] imidazolyl, imidazo [2',1':2,3] thiazolo [4,5-b ] pyrazinyl, imidazo [2',1':2,3] thiazolo [5,4-b ] pyridinyl, imidazo [2',1':2,3] thiazolo [4,5-c ] pyridyl, 1H-benzo [ f ] imidazo [4,5-b ] [1,4] thiazepinyl, and the like. Heteroaryl groups may be substituted with substituents described herein.
The terms "heterocyclyl", "heterocycle", "heteroalicyclic" or "heterocyclyl" are used interchangeably herein and all refer to monocyclic, bicyclic, tricyclic or tetracyclic ring systems wherein one or more atoms in the ring are independently optionally substituted with a heteroatom, the ring may be fully saturated or contain one or more degrees of unsaturation, but is by no means aromatic, and the heterocyclic ring system as a whole is not aromatic. For example, in some embodiments, bicyclic heterocyclic groups, wherein one ring has an aromatic characterAnd the other ring has no aromaticity as a whole. The heterocyclic ring system may be attached to the main structure at any heteroatom or carbon atom that results in the formation of a stable compound. One or more of the ring hydrogen atoms are independently substituted with one or more substituents described herein. Wherein some embodiments are "heterocyclyl", "heterocycle", "heteroalicyclic" or "heterocyclic" means a 4-12 membered monocyclic or polycyclic (3-9 carbon atoms and 1-3 heteroatoms selected from N, O, P, S, where S or P is independently optionally substituted with one or more oxygen atoms to give a compound like SO, SO 2 ，PO，PO 2 A group of (a) and (b) simultaneously, -CH 2 -the groups may independently optionally be replaced by-C (= O) -; when the ring is a three-membered ring, wherein there is only one heteroatom, when the ring is polycyclic, it may optionally be a bridged, spiro or fused ring). The term "C 3-9 The term "heterocyclic group" means a heterocyclic group having 3 to 9 carbon atoms, wherein the heterocyclic group has the above-mentioned meaning.
"Heterocyclyl" may be a carbon atom radical or a heteroatom radical. "Heterocyclyl" also includes heterocyclic groups fused to saturated or partially unsaturated rings or heterocycles. Examples of heterocycles include, but are not limited to, pyrrolidinyl, tetrahydrofuryl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, thiaxazanyl, azetidinyl, oxetanyl, thietanyl, piperidinyl, homopiperidinyl, epoxypropyl, azepinyl, oxepinyl, thietanyl, N-morpholinyl, 2-morpholinyl, 3-morpholinyl, thiomorpholinyl, N-piperazinyl, 2-piperazinyl, 3-piperazinyl, homopiperazinyl, 3534 zft 3534-tetrahydropyridin-1-yl, oxazepinyl, diazepinyl, thiazepinyl, pyrrolin-1-yl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2-indolinyl, 2H-pyranyl, 4H-pyranyl, dioxacyclohexyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithienoalkyl, dihydrothienyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,2,6-thiadiazinyl 1,1-dioxo-2-yl, hexahydro-2H- [1,4] dioxino [ 4232-c ] pyrrolyl, quinolizinyl, 1,1-thiomorpholinyl dioxide, 2,3,3a, 7a-tetrahydro-1H-isoindolyl, isoindolinyl, 1,2,3,4-tetrahydroquinolyl, dibenzofuranyl, dihydrobenzisothiazinyl, dihydrobenzisoxazinyl, dioxolanyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrazolyl, dihydropyrimidinyl, dihydropyrrolyl, 1,4-dithianyl, furanonyl, furanyl, imidazolidinyl, imidazolinyl, imidazolyl, imidazopyridinyl, imidazothiazolyl, indazolyl, indolinyl, indolizinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isobenzothienyl, isobenzodihydropyranyl, isocoumarinyl, isoindolyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, decahydroindolyl, oxadiazolyl, oxazolidinedionyl, oxazolidinyl, oxazolopyridinyl, oxiranyl, piperazinyl, piperidinyl, 4-piperidinonyl, pyrrolyl, quinuclidinyl, tetrahydroisoquinolinyl, tetrahydrothienyl, thiomorpholinyl, thiazolidinyl, 1,3,5-trithianyl, 2-oxopyrrolidinyl, oxo-63 zxft 3963-thiazolidinyl, 2-piperidino, 4325-dioxopiperidinyl, 3536 zxft 3736-dithiinyl, 2-oxodiketo 3926, bicyclo [ 395 ] morpholinyl, 2-oxodiketo, 3926, and bicyclo [ 3926.3 ] morpholinyl.
In some embodiments, heterocyclyl is a 4-12 atom heterocyclyl and refers to a saturated or partially unsaturated monocyclic or polycyclic ring containing 4-12 ring atoms wherein at least one ring atom is selected from the group consisting of nitrogen, sulfur, and oxygen atoms. Examples of heterocyclyl groups consisting of 4 to 12 atoms include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, 1,3-dioxolanyl, dithiocyclopentyl, tetrahydropyranyl, dihydropyranyl, 2H-pyranyl, 4H-pyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, dioxanyl, dithianyl, thiaoxanyl, homopiperazinyl, homopiperidinyl, oxepanyl, thietanyl, oxazepanyl, oxazepinyl, and thiazepinyl
Based on, diaza->
Based on the sulfur and nitrogen impurity->
Radicals, and the like. In heterocyclic radicals of-CH 2 Examples of-groups substituted by-C (= O) -include, but are not limited to, 2-oxopyrrolidinyl, oxo-1,3-thiazolidinyl, 2-piperidinonyl, 3,5-dioxopiperidinyl, pyrimidinedione, and the like. Examples of heterocyclic sulfur atoms that are oxidized include, but are not limited to, sulfolane, thiomorpholinyl 1,1-dioxide, and the like. Said heterocyclyl group of 4-7 atoms is optionally substituted with one or more substituents as described herein.
The term "heteroatom" refers to O, S, N, P and Si, including N, S and any oxidation state form of P; primary, secondary, tertiary amines and quaternary ammonium salt forms; or a form in which the hydrogen on a nitrogen atom in the heterocycle is substituted, e.g., N (like N in 3,4-dihydro-2H-pyrrolyl), NH (like NH in pyrrolidinyl) or NR (like NR in N-substituted pyrrolidinyl).
The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (1).
The terms "spirocyclic", "spiro", "spirobicyclic group", "spirobicyclic ring" indicate that one ring originates from a particular cyclic carbon on the other ring. For example, as described below, a saturated bridged ring system (rings D and B ') is referred to as a "fused bicyclic ring", whereas rings a' and D share a carbon atom in two saturated ring systems, and are referred to as "spiro rings". Each ring within the spiro ring is either carbocyclic or heterocyclic. Examples of such include, but are not limited to, 4-azaspiro [2.4] heptan-5-yl, 4-oxaspiro [2.4] heptan-5-yl, 5-azaspiro [2.4] heptan-5-yl, spiro [2.4] heptanyl, spiro [4.4] nonanyl, 7-hydroxy-5-azaspiro [2.4] heptan-5-yl, and the like.
The term "spiroheterobicyclic group" means that one ring originates from a specific cyclic carbon on the other ring. For example, as described above, a saturated bridged ring system (rings D and B ') is referred to as a "fused bicyclic ring", whereas rings a' and D share a carbon atom in two saturated ring systems, is referred to as a "spiro ring". And at least one ring system comprises one or more heteroatoms, wherein each ring system comprises 3 to 7 atoms, i.e. 1 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, S, whereby N, S or P independently optionally is substituted by one or more oxygen atoms to give compounds like NO, NO 2 ，SO，SO 2 ，PO，PO 2 A group of-CH 2 -the groups may independently optionally be replaced by-C (= O) -; examples of such include, but are not limited to, 4-azaspiro [2.4]]Heptalkyl, 4-oxaspiro [2.4]]Heptylalkyl, 5-azaspiro [2.4]Heptenyl, 2-azaspiro [4.5]]Decyl, 2-azaspiro [3.3]Heptylalkyl, 2-azaspiro [4.4]Nonanyl, 3-azaspiro [5.4 ]]Decyl, 2-oxo-6-azaspiro [3.3]Heptenyl, 2,6-diazaspiro [3.3]Heptylalkyl, 2-thio-6-azaspiro [3.3]Heptylalkyl 2-monoxides, 2-thio-6-azaspiro [3.3]Heptylalkyl 2,2-dioxide, 2,8-diazaspiro [4.5]]Decyl, 2,7-diazaspiro [4.4]]Octyl 2,7-diazaspiro [4.5]]Decyl, 2,6-diazaspiro [4.5]Decyl, 2,8-diazaspiro [4.5]Decan-3-one-yl, 1,8-diazaspiro [4.5]Decyl, 1,7-diazaspiro [4.4]]Nonanyl, 1,7-diazaspiro [4.4]]Nonan-6-one-yl, 2,9-diazaspiro [5.5]]Undec-1-on-yl, 1-oxo-3,8-diazaspiro [4.5]Decan-2-one-yl, 1-oxo-3,7-diazaspiro [4.5]Decan-2-one-yl, 2,6-diazaspiro [3.4]Octyl 2,5-diazaspiro [3.5]]Nonanyl, 2,6-diazaspiro [3.3]Heptylalkyl, 2-oxo-7-azaspiro [3.5]Nonanyl, 2-oxo-6-azaspiro [3.4]]An octyl group and the like. The spiroheterocyclyl group may be substituted with the substituents described herein.
The terms "fused bicyclic", "fused ring", "fused bicyclic group" or "fused ring group" mean a fused ring system, saturated or unsaturated, involving a non-aromatic bicyclic ring system, at least one ring of which is non-aromatic. Such systems may contain independent or conjugated unsaturation, but the core structure does not contain aromatic or heteroaromatic rings (although aromatics may be substituted thereon). Examples of fused bicyclic rings in which each ring is either a carbocyclic or a heteroalicyclic include, but are not limited to, hexahydro-furan [3,2-b ] furanyl, 2, 3a,4,7 a-hexahydro-1H-indenyl, 7-azabicyclo [2.2.1] heptanyl, fused bicyclo [3.3.0] octanyl, fused bicyclo [3.1.0] hexanyl, 1,2,3, 4a,5,8 a-octahydronaphthyl, all of which are included within the fused bicyclic ring system.
The term "fused heterobicyclic group" refers to a saturated or unsaturated fused ring system, involving a non-aromatic bicyclic ring system, at least one of the rings being non-aromatic. Such systems may contain independent or conjugated unsaturation, but the core structure does not contain aromatic or heteroaromatic rings (although aromatics may be substituents thereon). And at least one ring system comprises one or more heteroatoms, wherein each ring system comprises a ring of 3 to 7 atoms, i.e. 1 to 6 carbon atoms and 1 to 3 heteroatoms selected from N, O, P, S, wherein N, S or P independently optionally is substituted by one or more oxygen atoms to give compounds like NO, NO 2 ，SO，SO 2 ，PO，PO 2 A group of-CH 2 The-groups may independently optionally be replaced by-C (= O) -examples of which include, but are not limited to, hexahydro-2H- [1,4]Dioxin [2,3-c]) Pyrrolyl, 3-azabicyclo [3.3.0]Octyl, 8-azabicyclo [4.3.0 ] groups]Nonanyl, 8-azabicyclo [4.3.0]Nonane 3-yl, 3-azabicyclo [4.3.0]Nonan-3-yl, 1,5-dioxo-8-azabicyclo [4.3.0]Nonyl, (1R, 6S) -2,5-dioxo-8-azabicyclo [4.3.0]Nonyl, (1R, 6R) -2,5-dioxo-8-azabicyclo [4.3.0]Nonanyl, isoindolinyl, 1,2,3,4-tetrahydroquinolinyl, (1S, 5S) -1-hydroxy-3-azabicyclo [3.1.0]Hexane radical, (1R, 5S) -1-hydroxy-3-azabicyclo [3.1.0]Hexane radical, 3-aza-7-oxabicyclo [3.3.0]Octyl 3,7-diazabicyclo [3.3.0]Octyl 2,6-diazabicyclo [3.3.0]Octyl, 2,7-diazabicyclo [3.3.0]An octyl group, a vinyl group and a vinyl group,2,7-diazabicyclo [3.3.0]Octyl, 2,8-diazabicyclo [4.3.0 ]]Nonanyl, 3,8-diazabicyclo [4.3.0]Nonanyl, 3-oxo-8-azabicyclo [4.3.0]Nonanyl, 2-oxo-8-azabicyclo [4.3.0]Nonanyl, 2,8-diaza-5-oxabicyclo [4.3.0]Nonanyl, 4,9-diazabicyclo [4.3.0 ]]Nonanyl, 2,9-diazabicyclo [4.3.0]Nonanyl, 2-oxo-3-oxo-8-azabicyclo [4.3.0]Nonanyl, 3-oxo-2,4,8-triazabicyclo [4.3.0]Nonanyl, 3-oxo-4-oxo-2,8-diazabicyclo [4.3.0]Nonanyl, 3-oxo-2,8-diazabicyclo [4.3.0]Nonanyl, 3,8-diazabicyclo [4.3.0 ]]Nonanyl, 3,7-diazabicyclo [4.3.0 ]]Nonanyl, 3,9-diazabicyclo [4.3.0 ]]Nonanyl, 3-oxo-8-azabicyclo [4.3.0]Nonanyl, 3-thio-8-azabicyclo [4.3.0]Nonanyl, 5,6-dihydro-4H-pyrrolo [3,4-c]Isoxazolyl, [1,2,4]Triazole [4,3-a]Piperidinyl, isoxazolo [4,3-c]Piperidinyl, 4,5,6,7-tetrahydroisooxazolo [3,4-c]Pyridyl, [1,2,4]Triazole [4,3-a]Piperazinyl, 2-oxo-3-oxo-8-azabicyclo [4.3.0]Nonanyl, 2-oxo-7-azabicyclo [4.4.0]Decyl, 1,5-dioxo-9-azabicyclo [4.4.0]Decyl, 3-azabicyclo [4.4.0]Decyl, 2,7-diazanaphthyl, 2-oxo-8-azabicyclo [4.4.0]Decyl, hexahydropyrrolo [1,2-a]Pyrazin-1 (2H) -one-yl, decahydro-1H-pyrido [1,2-a]Pyrazin-1-one-yl, 3-azabicyclo [3,1,0]Hexane-1-amino-yl, and the like. The fused heterobicyclic group may be substituted with the substituent described in the present invention.
As described herein, the substituent is attached to the ring through a bond to form a ring system, which means that the substituent can be substituted at any substitutable position on the ring. For example, formula (a) represents that the p substituents R may be substituted at any possible substituted position on the pyridine ring.
As described herein, the ring system formed by the attachment of a linker to the ring (as shown in formula b) represents that the linker may be attached to the rest of the molecule at any available position on the ring system. Formula b represents that any possible linking position on the octahydro-cyclopentenopyrrole ring can be linked to the rest of the molecule.
As described herein, there are two attachment points on ring C that can be attached to the rest of the molecule, for example, as shown in formula C, where either the E or E' terminus in formula C is attached to the rest of the molecule, i.e., the attachment of the two ends can be interchanged, unless explicitly stated otherwise.
The point of attachment may be attached to the rest of the molecule at any point on the ring that is attachable, as described herein, while the two ends of the attachment may be interchanged. For example, formula d represents any possible attachment location on the ring as a point of attachment, with the two ends of the attachment point being interchangeable.
In addition, it should be noted that, unless explicitly stated otherwise, the description taken throughout this document "each.. And.. Independently" and "each.. And.. Each independently" and.. Each.. Independently "may be used interchangeably and should be understood broadly to mean that the particular options expressed between the same symbols in different groups do not affect each other, or that the particular options expressed between the same symbols in the same groups do not affect each other. For example,
wherein N1 and N2 represent the same or different atoms or groups and do not affect each other; "n1-n8" represent the same or different values and do not affect each other.
The term "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastrointestinal distress, dizziness and the like, when administered to a human. Preferably, the term "pharmaceutically acceptable" as used herein refers to those approved by a federal regulatory agency or a state government or listed on the U.S. pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.
The term "carrier" refers to a diluent, adjuvant, excipient, or matrix with which the compound is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Aqueous saline solutions and aqueous dextrose and glycerol solutions are preferably used as carriers, particularly injectable solutions. Suitable Pharmaceutical carriers are described in e.w. martin, "Remington's Pharmaceutical Sciences".
The "hydrate" of the present invention refers to the compound provided by the present invention or a salt thereof, which further comprises a stoichiometric amount or a non-stoichiometric equivalent of water bound by non-covalent intermolecular forces, and may be said to be an association of solvent molecules with water.
"solvate" of the present invention refers to an association of one or more solvent molecules with a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, meOH, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol.
The "ester" of the present invention means that the compounds represented by the formulae (I) to (II) having a hydroxyl group can form an in vivo hydrolysable ester. Such esters are, for example, pharmaceutically acceptable esters which are hydrolysed in the human or animal body to yield the parent alcohol. The group of the in vivo hydrolysable ester of the compounds of formula (I) -formula (II) containing a hydroxyl group includes, but is not limited to, phosphate, acetoxymethoxy, 2,2-dimethylpropionyloxymethoxy, alkanoyl, benzoyl, benzoylacetyl, alkoxycarbonyl, dialkylcarbamoyl and N- (dialkylaminoethyl) -N-alkylcarbamoyl.
By "nitroxide" herein is meant that when a compound contains several amine functional groups, 1 or more than 1 nitrogen atom can be oxidized to form an N-oxide. Specific examples of N-oxides are N-oxides of tertiary amines or N-oxides of nitrogen-containing heterocyclic nitrogen atoms. The corresponding amines can be treated with an oxidizing agent such as hydrogen peroxide or a peracid (e.g., peroxycarboxylic acid) to form the N-oxide (see Advanced Organic Chemistry, wiley Interscience, 4 th edition, jerry March, pages). In particular, the N-oxide may be prepared by the method of l.w. ready (syn.comm.1977, 7, 509-514) in which an amine compound is reacted with m-chloroperoxybenzoic acid (MCPBA), for example, in an inert solvent such as DCM.
The compounds may exist in a number of different geometric isomers and tautomers and the compounds of formula (I) -formula (II) include all such forms. For the avoidance of doubt, when a compound exists as one of several geometric isomers or tautomers and only one is specifically described or shown, it is clear that all other forms are included in formula (I) -formula (II).
The term "prodrug", as used herein, represents a compound that is converted in vivo to a compound of formula (I) -formula (II). Such conversion is effected by hydrolysis of the prodrug in the blood or by enzymatic conversion to the parent structure in the blood or tissue. The prodrug compound of the invention can be ester, and in the prior invention, the ester can be used as the prodrug and can be phenyl ester, aliphatic (C) 1-24 ) Esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters. For example, a compound of the present invention contains a hydroxy group, i.e., it can be acylated to provide the compound in prodrug form. Other prodrug forms include phosphate esters, such as those phosphate esters which are phosphorylated via the parent hydroxy group. For a complete discussion of prodrugs, reference may be made to the following: t.higuchi and v.stella, pro-drugs as Novel Delivery Systems, vol.14 of the a.c.s.symposium Series, edward b.roche, ed., bioreversible Carriers in Drug Design, american Pharmaceutical Association and Pergamon Press,1987, j.rautio et al, produgs: design and Clinical Applications, nature Review Drug Discovery,2008,7, 255-270, and S.J.Hecker et al, drivers of Phosphonates and Phosphonates, journal of medical chemistry,2008, 51, 2328-2345。
Unless otherwise indicated, all tautomeric forms of the compounds of the invention are included within the scope of the invention.
In addition, unless otherwise indicated, the structural formulae of the compounds described herein include isotopically enriched concentrations of one or more different atoms. The invention includes isotopically-labeled compounds, which are identical to those recited in formula (I) -formula (II), but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as 2 H、 3 H、 13 C、 11 C、 14 C、 15 N、 18 O、 17 O、 31 P、 32 P、 35 S、 18 F and 36 and (4) Cl. Compounds of the present invention, prodrugs thereof, and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically-labelled compounds of the invention, e.g. by incorporation of radioactive isotopes (e.g. by introducing 3 H and 14 c) Can be used in drug and/or substrate tissue distribution assays. Isotopically labeled compounds of formula (I) -formula (II) of the present invention and prodrugs thereof can generally be prepared by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent in carrying out the procedures disclosed in the schemes and/or in the examples and preparations below.
"metabolite" refers to the product of a particular compound or salt thereof obtained by metabolism in vivo. Metabolites of a compound can be identified by techniques well known in the art, and its activity can be characterized by assays as described herein. Such products may be obtained by administering the compound by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic cleavage, and the like. Accordingly, the present invention includes metabolites of compounds, including metabolites produced by contacting a compound of the present invention with a mammal for a sufficient period of time.
Various pharmaceutically acceptable salt forms of the compounds of the invention are useful. The term "pharmaceutically acceptable salts" means those salt forms that are readily apparent to the pharmaceutical chemist as being substantially non-toxic and providing the desired pharmacokinetic properties, palatability, absorption, distribution, metabolism or excretion. Other factors, more practical in nature, are also important for selection, these are: cost of raw materials, ease of crystallization, yield, stability, hygroscopicity and, as a result, flowability of the drug substance. Briefly, the pharmaceutical composition may be prepared by combining the active ingredient with a pharmaceutically acceptable carrier.
As used herein, "pharmaceutically acceptable salts" refers to both organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as are: s.m. Berge et al, description pharmacological acceptable salts in detail in j. Pharmaceutical Sciences,66:1-19, 1977. Pharmaceutically acceptable non-toxic acid salts include, but are not limited to, inorganic acid salts formed by reaction with amino groups such as hydrochloride, hydrobromide, phosphate, sulfate, perchlorate, nitrate and the like, and organic acid salts such as acetate, propionate, glycolate, oxalate, maleate, malonate, succinate, fumarate, tartrate, citrate, benzoate, mandelate, methanesulfonate, ethanesulfonate, toluenesulfonate, sulfosalicylate and the like, or obtained by other methods described in the literature such as ion exchange.
Other pharmaceutically acceptable salts include adipates, malates, 2-hydroxypropionic acid, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, cyclopentylpropionates, digluconates, dodecylsulfates, ethanesulfonates, formates, fumarates, glucoheptonates, glycerophosphates, gluconates, hemisulfatesSalts, heptanoates, hexanoates, hydroiodides, 2-hydroxy-ethanesulfonates, lactobionates, lactates, laurates, lauryl sulfates, malates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, palmitates, embonates, pectates, persulfates, 3-phenylpropionates, picrates, pivalates, propionates, stearates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, and the like. Salts obtained with appropriate bases include alkali metals, alkaline earth metals, ammonium and N + (C 1-4 Alkyl radical) 4 A salt.
The present invention also contemplates quaternary ammonium salts formed from any compound containing a group of N. Water-soluble or oil-soluble or dispersion products can be obtained by quaternization. The alkali metal or alkaline earth metal salt includes sodium salt, lithium salt, potassium salt, calcium salt, magnesium salt, iron salt, zinc salt, copper salt, manganese salt, aluminum salt and the like. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts and amine cations resistant to formation of counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C 1-8 Sulfonates and aromatic sulfonates. Amine salts such as, but not limited to, N '-dibenzylethylenediamine, chloroprocaine, choline, ammonia, diethanolamine and other hydroxyalkylamines, ethylenediamine, N-methyl reduced glucamine, procaine, N-benzylphenethylamine, 1-p-chlorobenzyl-2-pyrrolidin-1' -ylmethyl-benzimidazole, diethylamine and other alkylamines, piperazine and tris (hydroxymethyl) aminomethane; alkaline earth metal salts such as, but not limited to, barium, calcium and magnesium; a transition metal salt such as, but not limited to, zinc.
The term "protecting group" or "Pg" refers to a substituent that when reacted with another functional group, is typically used to block or protect a particular functionality. For example, "amino protecting group" refers to a substituent attached to an amino group to block or protect the functionality of the amino group in a compound, and suitable amino protecting groups include acetyl, trifluoroacetyl, t-Butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9-fluorenylmethylenoxycarbonyl (Fmoc). Similarly, "hydroxy protecting group" meansSubstituents for the hydroxyl group serve to block or protect the functionality of the hydroxyl group, and suitable protecting groups include acetyl and silyl groups. "carboxy protecting group" refers to the functionality of a substituent of a carboxy group to block or protect the carboxy group, and typical carboxy protecting groups include-CH 2 CH 2 SO 2 Ph, cyanoethyl, 2- (trimethylsilyl) ethyl, 2- (trimethylsilyl) ethoxymethyl, 2- (p-toluenesulfonyl) ethyl, 2- (p-nitrobenzenesulfonyl) ethyl, 2- (diphenylphosphino) ethyl, nitroethyl, and the like. General descriptions of protecting groups can be found in the literature: greene, protective Groups in Organic Synthesis, john Wiley&Sons，New York，1991；and P.J.Kocienski，Protecting Groups，Thieme，Stuttgart，2005.
The term "room temperature" as used herein means a temperature of from 10 ℃ to 40 ℃. In some embodiments, "room temperature" refers to a temperature from 20 ℃ to 30 ℃; in other embodiments, "room temperature" refers to 25 ℃.
In this specification, a structure is dominant if there is any difference between the chemical name and the chemical structure.
Abbreviations for any protecting groups, amino acids and other compounds used in the present invention shall, unless otherwise indicated, be referred to their commonly used, accepted abbreviations or refer to the IUPAC-IUB Commission on Biochemical Nomenclature (see biochem.1972, 11, 942-944).
Description of the Compounds of the invention
The invention provides a compound or a pharmaceutical composition thereof, which can be used as an inhibitor of PD-1/PD-L1. The invention further relates to the use of said compounds or pharmaceutical compositions thereof for the preparation of a medicament for the treatment of diseases and/or disorders by inhibiting PD-1 activity with said compounds. The invention further describes methods for synthesizing the compounds. The compounds of the invention show improved biological activity and pharmacokinetic properties.
The invention provides a compound of a PD-1/PD-L1 small molecule inhibitor, which has a structure shown in a formula (I) or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, an ester, a pharmaceutically acceptable salt or a prodrug of the structure shown in the formula (I),
wherein A, X, R 1 、R 2 、R 3 And p has the meaning described in the present invention.
In some embodiments, R is as described herein 1 Is- (CH) 2 ) n Ar, wherein Ar is C 6-12 Aryl or C 5-12 A heteroaryl group; said Ar is optionally substituted with 1,2,3 or 4 substituents each independently being H, D, cyano, halogen, amino, methanesulfonyl, acetamido, C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or C 1-6 An alkoxy group; n is 1,2,3 or 4.
In some embodiments, R as described herein 1 Is- (CH) 2 ) n Ar, wherein Ar is phenyl, pyridyl, pyrimidyl, indolyl or quinolyl; ar is optionally substituted with 1,2,3 or 4 substituents each independently being H, cyano, halogen, amino, methanesulfonyl, acetylamino, C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy or C 1-6 An alkoxy group; n is 1.
In some embodiments, A is-CH 2 O-、-OCH 2 -、-CH 2 -CH 2 -, -C (O) NH-or-NHC (O) -.
In some embodiments, A is-CH 2 O-。
In some embodiments, R is as described herein 2 Is composed of
Wherein R is 5 And R 6 Each independently of the other being H, halogen, C 1-6 Alkyl radical, C 1-6 Haloalkyl or C 1-6 An alkoxy group; z is CH or N;
R 7 is composed of
q is 1,2,3 or 4;
R v is composed of
Wherein R is v Optionally substituted with 0,1, 2 or 3 substituents selected from hydroxy, cyano, amino, oxo (= O), C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy or amino C 1-6 An alkyl group; w is a group of 1 、W 3 And W 5 Each independently is CH 2 、S、O、S(O) 2 Or NH, W 2 And W 4 Each independently is CH or N; m1, m2, m3, m4, m5, m6, m7 and m8 are each independently 0,1, 2 or 3;
x and Y are H, C 1-6 Alkyl radical, C 1-6 Alkoxy, halogen or cyano.
In some embodiments, R is as described herein 5 Or R 6 Each independently is H, D, fluoro, chloro, bromo, methyl, ethyl, n-propyl, isopropyl, tert-butyl, isobutyl, n-butyl, trifluoromethyl, difluoromethyl, monofluoromethyl, methoxy, or ethoxy.
In some embodiments, R is as described herein 7 Is composed of
q is 1.
In some embodiments, R as described herein v Is composed of
Wherein R is v Optionally substituted with 0,1, 2 or 3 substituents selected from hydroxy, cyano, amino, oxo (= O), C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy or amino C 1-6 An alkyl group; w 1 、W 3 And W 5 Each independently is CH 2 、S、O、S(O) 2 Or NH, W 2 And W 4 Each independently is CH or N; m1, m2, m3, m4, m5, m6, m7 and m8 are each independently 0,1, 2 or 3.
In some embodiments, R is as described herein v Is one of the groups formed by the following structural formula:
wherein, the W 1 、W 3 And W 5 Each independently is CH 2 、S、O、S(O) 2 Or NH, W 2 And W 4 Each independently is CH or N; m1, m2, m3, m4, m5, m6, m7 and m8 are each independently 0,1, 2 or 3;
the R is v Optionally substituted with 0,1, 2 or 3 substituents selected from hydroxy, cyano, amino, oxo (= O), C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy or amino C 1-6 An alkyl group.
In other embodiments, R is as described herein v Is one of the groups formed by the following structural formula:
wherein, R is v Optionally substituted by 0,1, 2Or 3 substituents, said substituents being hydroxy, cyano, amino, oxo (= O), C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy or amino C 1-6 An alkyl group.
In some embodiments, Y is methyl, ethyl, isopropyl, n-propyl, n-butyl, t-butyl, or isobutyl; preferably, Y is methyl.
In some embodiments, R as described herein 3 Is C 1-8 Alkylamino or C 3-9 Heterocyclic group, wherein, said C 3-9 The heterocyclic group contains at least one N atom; said C 1-8 Alkylamino or C 3-9 The heterocyclyl is optionally substituted with 0,1, 2,3 or 4 substituents selected from hydrogen, hydroxy, halogen, carboxy, C 1-6 Alkyl radical, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 1-6 Alkoxy, amino C 1-6 Alkyl, acetylamino, cyano, sulfonamido or oxo (= O).
In some embodiments, R3 according to the present invention is one of the groups formed by the following structures:
in some embodiments, p is 0,1, 2, or 3.
In some embodiments, X is H, C as described herein 1-6 Alkyl radical, C 1-6 Alkoxy, halogen or cyano; preferably, X is fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl.
In some embodiments, the compounds of the invention are stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, esters, pharmaceutically acceptable salts or prodrugs of the structures of formula (II) or of the structures of formula (II),
wherein R is 1 、R 2 、R 3 、R 5 、R 6 、R v X, Y, Z and p have the meaning described in the present invention.
In some embodiments, the present invention comprises a compound of one of the following or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, pharmaceutically acceptable salt or prodrug of a compound of one of the following:
in one aspect, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) or formula (II) of the present invention, or a stereoisomer, a geometric isomer, a tautomer, a nitrogen oxide, a hydrate, a solvate, a metabolite, a pharmaceutically acceptable salt, or a prodrug thereof, and a pharmaceutically acceptable adjuvant, or a combination thereof.
In one aspect, the invention relates to the use of a compound of formula (I) or (II) or a pharmaceutical composition thereof for the manufacture of a medicament for the prevention, treatment or alleviation of a disease or a disorder associated with the PD-1/PD-L1 signalling pathway in a patient.
In some embodiments, the disease associated with the PD-1/PD-L1 signaling pathway described herein is cancer, an infectious disease, or an autoimmune disease.
In other embodiments, the cancer of the invention is selected from the group consisting of diseases in which there is unlimited proliferation of cells in an organ or body tissue; the infectious diseases are bacterial infectious diseases, virus infectious diseases or fungal infectious diseases; the autoimmune disease is an organ-specific autoimmune disease or a systemic autoimmune disease.
In some embodiments, the disease of unlimited proliferation of cells in an organ or body tissue described herein is bone cancer, head and neck cancer, pancreatic cancer, skin cancer, malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, hodgkin's disease, non-hodgkin's lymphoma, carcinoma of the esophagus, carcinoma of the small intestine, cancer of the endocrine system, carcinoma of the thyroid gland, carcinoma of the parathyroid gland, carcinoma of the adrenal gland, sarcoma of soft tissue, carcinoma of the urethra, carcinoma of the penis, chronic or acute leukemia, solid tumors of childhood, lymphocytic lymphomas, carcinoma of the bladder, carcinoma of the kidney or ureter, carcinoma of the renal pelvis, tumors of the Central Nervous System (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumors, brain stem glioma, pituitary adenoma, kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, or an environmentally induced cancer or a combination of the above; wherein the chronic or acute leukemia comprises acute myelocytic leukemia, chronic myelocytic leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia.
In some embodiments, the viral infectious disease of the present invention is aids, hepatitis a, hepatitis b, hepatitis c, hepatitis d, herpes virus infection, papillomavirus infection, and influenza.
In some embodiments, the organ-specific autoimmune disease described herein is chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis, and acute idiopathic polyneuritis; the systemic autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease and autoimmune hemolytic anemia.
In some embodiments, the invention features a method of protecting, managing, treating, or ameliorating a disease associated with the PD-1/PD-L1 signaling pathway in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound or pharmaceutical composition of the invention.
In some embodiments, the disease associated with the PD-1/PD-L1 signaling pathway described herein is cancer, an infectious disease, or an autoimmune disease.
In some embodiments, the cancer of the invention is a disease in which there is unlimited proliferation of cells in an organ or a body tissue; the infectious diseases are bacterial infectious diseases, virus infectious diseases or fungal infectious diseases; the autoimmune disease is an organ-specific autoimmune disease or a systemic autoimmune disease.
In some embodiments, the disease of unlimited proliferation of cells in an organ or body tissue described herein is bone cancer, head and neck cancer, pancreatic cancer, skin cancer, malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, hodgkin's disease, non-hodgkin's lymphoma, carcinoma of the esophagus, carcinoma of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemia, solid tumor of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, central Nervous System (CNS) tumor, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, or an environmentally-induced cancer, or a combination of the foregoing; wherein the chronic or acute leukemia comprises acute myelocytic leukemia, chronic myelocytic leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia.
In some embodiments, the viral infectious disease described herein is aids, hepatitis a, hepatitis b, hepatitis c, hepatitis d, herpes virus infection, papillomavirus infection, and influenza.
In some embodiments, the organ-specific immune disease of the invention is chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis, and acute idiopathic polyneuritis; the systemic autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease and autoimmune hemolytic anemia. In another aspect, the invention provides a method of modulating an immune response mediated by the PD-1 signaling pathway in a subject, comprising administering to the subject a therapeutically effective amount of a compound of the invention, thereby modulating the immune response in the subject.
In some embodiments, the compounds or pharmaceutical compositions described herein are used to protect, manage, treat, or ameliorate a disease associated with the PD-1/PD-L1 signaling pathway in a patient.
In some embodiments, the disease associated with the PD-1/PD-L1 signaling pathway described herein is cancer, an infectious disease, or an autoimmune disease.
In some embodiments, the cancer of the invention is a disease in which there is unlimited proliferation of cells in an organ or a body tissue; the infectious diseases are bacterial infectious diseases, virus infectious diseases or fungal infectious diseases; the autoimmune disease is an organ-specific autoimmune disease or a systemic autoimmune disease.
In some embodiments, the disease of unlimited proliferation of cells in an organ or body tissue described herein is bone cancer, head and neck cancer, pancreatic cancer, skin cancer, malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, hodgkin's disease, non-hodgkin's lymphoma, carcinoma of the esophagus, carcinoma of the small intestine, cancer of the endocrine system, carcinoma of the thyroid gland, carcinoma of the parathyroid gland, carcinoma of the adrenal gland, sarcoma of soft tissue, carcinoma of the urethra, carcinoma of the penis, chronic or acute leukemia, solid tumor of the child, lymphocytic lymphomas, carcinoma of the bladder, carcinoma of the kidney or ureter, carcinoma of the renal pelvis, tumor of the Central Nervous System (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, or an environmentally induced cancer or a combination of the foregoing; wherein the chronic or acute leukemia comprises acute myelocytic leukemia, chronic myelocytic leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia.
In some embodiments, the viral infectious disease described herein is aids, hepatitis a, hepatitis b, hepatitis c, hepatitis d, herpes virus infection, papillomavirus infection, and influenza.
In some embodiments, the organ-specific immune disease of the invention is chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis, and acute idiopathic polyneuritis; the systemic autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease and autoimmune hemolytic anemia.
In another aspect, the invention provides a method of modulating an immune response in a subject mediated by the PD-1 signaling pathway, comprising administering to the subject a therapeutically effective amount of a compound of the invention, thereby modulating the immune response in the subject.
In another aspect, the invention relates to methods for the preparation, isolation and purification of compounds encompassed by formula (I) or (II).
Compositions of the compounds of the invention
As described herein, the pharmaceutical composition of the present invention comprises any one of the compounds of formula (I) or (II) of the present invention, and further comprises pharmaceutically acceptable excipients, such as any solvent, solid excipient, diluent, binder, disintegrant, or other liquid excipient, dispersant, flavoring agent or suspending agent, surfactant, isotonic agent, thickening agent, emulsifier, preservative, solid binder or lubricant, and the like, as used herein, suitable for the particular intended dosage form. As described in the following documents: in Remington: the Science and Practice of Pharmacy,21st edition,2005, ed.D.B.Troy, lippincott Williams and Wilkins, philadelphia, and Encyclopedia of Pharmaceutical technology, eds.J.Swarbrick and J.C.Boylan,1988-1999.Marcel Dekker, new York, taken together with The disclosure of The literature herein, indicates that different excipients can be used in The formulation of pharmaceutically acceptable compositions and their well-known methods of preparation. Except insofar as any conventional adjuvant is incompatible with the compounds of the invention, e.g., any adverse biological effect produced or interaction in a deleterious manner with any other component of a pharmaceutically acceptable composition, their use is contemplated by the present invention.
Substances that may serve as pharmaceutically acceptable excipients include, but are not limited to, ion exchangers; aluminum; aluminum stearate; lecithin; serum proteins, such as human serum albumin; buffer substances such as phosphates; glycine; sorbic acid; potassium sorbate; a mixture of partial glycerides of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts; colloidal silica; magnesium trisilicate; polyvinylpyrrolidone; polyacrylate esters; a wax; polyethylene-polyoxypropylene-blocking polymers; lanolin; sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; gum powder; malt; gelatin; talc powder; adjuvants such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic salt; ringer's solution; ethanol; phosphoric acid buffer solution; and other non-toxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate; a colorant; a release agent; coating the coating material; a sweetener; a flavoring agent; a fragrance; preservatives and antioxidants. When the compound of the present invention is administered in the form of a medicament to a mammal such as a human, it may be administered in the form of the compound itself or may be administered in the form of a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably 0.5 to 90%) of an active ingredient and a pharmaceutically acceptable carrier.
By "therapeutically effective amount" or "effective amount" is meant a sufficient amount of one or more compounds of the invention to treat or prevent a particular disease, disorder, or syndrome, to alleviate, ameliorate, or eliminate one or more symptoms of the particular disease, disorder, or syndrome, or to prevent or delay the onset of one or more symptoms of the particular disease, disorder, or syndrome described herein. In the case of cancer, a therapeutically effective amount of the drug may reduce the number of cancer cells; reducing the size of the cancer; inhibit (i.e., slow to some extent or alternatively stop) cancer cell infiltration into peripheral organs; suppress (i.e., slow to some extent or alternatively stop) tumor metastasis; tumor growth has continued to a certain extent; and/or relieve to some extent one or more symptoms associated with cancer. In the case of infectious disease states, a therapeutically effective amount is an amount sufficient to reduce or alleviate infectious disease (symptoms of infection caused by bacteria, viruses, and fungi). One of ordinary skill in the art will be able to study the factors contained herein and determine, without undue experimentation, an effective amount of a compound of the invention.
The administration regimen may affect the constitution of the effective amount. The compounds of the invention may be administered to an individual prior to or after the onset of a condition associated with the PD-1/PD-L1 signaling pathway. In addition, multiple divided doses and staggered doses may be administered daily or sequentially, or may be administered as a continuous infusion, or may be administered as a bolus. Furthermore, the dosage of the compounds of the invention may be increased or decreased as appropriate, depending on the exigencies of the condition being treated or prevented.
The compounds of the invention are useful for treating the conditions, disorders or diseases described herein, or for the preparation of pharmaceutical compositions for treating such diseases. Methods of use of the compounds of the invention in the treatment of such diseases or pharmaceutical formulations containing the compounds of the invention for use in the treatment of such diseases.
"pharmaceutically acceptable carriers" are art-recognized and include pharmaceutically acceptable materials, compositions or vehicles suitable for administration of the compounds of the invention to a mammal. The carrier may include a liquid or solid filler, diluent, excipient, solvent or encapsulating material which is involved in carrying the subject substance or in transferring it from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: sugars such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc powder; excipients, such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline water; ringer's solution; ethanol; phosphate buffer; and other non-toxic compatible materials used in pharmaceutical formulations.
Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition.
Examples of pharmaceutically acceptable antioxidants include: water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; oil-soluble antioxidants such as ascorbyl palmitate, butylated Hydroxyanisole (BHA), butylated Hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.
Formulations of the present invention include those suitable for oral, nasal, topical, buccal, sublingual, rectal, vaginal and/or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form is generally that amount of the compound which produces a therapeutic effect. Generally, the amount is from about 1% to about 99% active ingredient, preferably from about 5% to about 70%, most preferably from about 10 to about 30%, in units of one percent.
The methods of making these formulations or compositions include the step of bringing into association a compound of the invention with a carrier, optionally and independently one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the compounds of the invention with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.
Formulations of the present invention suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored base, usually sucrose and acacia or tragacanth), powders, granules, or solutions or suspensions in aqueous or non-aqueous liquids, or oil-in-water or water-in-oil liquid emulsions, or elixirs or syrups, or pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or mouthwashes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. The compounds of the invention may also be administered in the form of a bolus, electuary or paste.
In solid dosage forms of the invention (capsules, tablets, pills, dragees, powders, granules and the like) for oral administration, the active ingredient is mixed with one or more pharmaceutically acceptable carriers such as sodium citrate or dicalcium phosphate and/or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; binders, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicic acids and sodium carbonate; solution retarding agents (solution retading agents), such as paraffin; absorption promoters, such as quaternary ammonium compounds; wetting agents, for example, cetyl alcohol and glycerol monostearate; adsorbents such as kaolin and bentonite; lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and a colorant. In the case of capsules, tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
Tablets may be prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared with binders (e.g., gelatin or hydroxypropylmethyl cellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surfactants or dispersing agents. Molded tablets may be prepared by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.
Tablets and other solid dosage forms of the pharmaceutical compositions of the invention, such as dragees, capsules, pills and granules, can optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical art. They may also be formulated with, for example, hydroxypropylmethyl cellulose in various proportions to provide the desired release properties, other polymer matrices, liposomes and/or microspheres to provide slow or controlled release of the active ingredient therein. They may be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water or some other injectable sterile vehicle immediately prior to use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient may also be in microencapsulated form, if appropriate with the use of one or more of the above-mentioned excipients.
Liquid dosage forms of the compounds of the invention for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredients, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
In addition to inert diluents, the oral compositions can also include adjuvants (adjuvants) such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
Suspensions, in addition to the active compounds, may contain co-excipients, for example ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.
Formulations of the pharmaceutical compositions of the present invention for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the present invention with one or more suitable non-irritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or salicylate, and which is solid at room temperature but liquid at body temperature and will therefore melt in the rectum or vaginal cavity and release the active compound.
Formulations of the invention suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.
Dosage forms for topical or transdermal administration of the compounds of the invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active ingredient may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants which may be required.
Ointments, pastes, creams and gels may contain, in addition to an active compound of the invention, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.
Powders and sprays can contain, in addition to a compound of the invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder or mixtures of these substances. Sprays can also contain conventional propellants such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.
Transdermal patches have the additional advantage of providing controlled delivery of the compounds of the present invention to the body. Such dosage forms may be prepared by dissolving or dispersing the compound in a suitable vehicle. Absorption enhancers may also be used to increase the flux of compounds through the skin. The rate of such flow can be controlled by providing a rate controlling membrane or dispersing the active compound in a polymer matrix or gel.
Also included within the scope of the present invention are ophthalmic formulations, ophthalmic ointments, powders, solutions, and the like.
Pharmaceutical compositions of the invention suitable for parenteral administration comprise one or more compounds of the invention and one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions immediately prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the recipient or suspending or thickening agents.
Examples of suitable aqueous and nonaqueous carriers that can be used in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Suitable fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.
These compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like in the composition. In addition, prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.
In some cases, in order to prolong the effect of the drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material which is poorly water soluble. Thus, the rate of absorption of the drug will depend on its rate of dissolution, which in turn may depend on crystal size and crystal form. Alternatively, prolonged absorption of a parenterally administered drug form is achieved by dissolving or suspending the drug in an oily matrix.
Injectable depot forms are prepared by forming microencapsule matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular compound used, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Injectable depot formulations can also be prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.
The formulations of the present invention may be administered orally, parenterally, topically or rectally. They are of course administered in a form suitable for the respective route of administration. For example, they are administered in the form of tablets or capsules, by injection, inhalation, ophthalmic lotion, ointment, suppository and the like, by injection, infusion or inhalation; topically applied by lotion or ointment; administered rectally by means of suppositories. Oral and/or intravenous administration is preferred.
The phrase "parenteral administration" as used herein means modes of administration other than enteral and topical administration, typically by injection, including, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.
The terms "systemic administration" and "peripheral administration" as used herein mean administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the patient's system and is thus subject to metabolism and other similar processes, e.g., subcutaneous administration.
These compounds may be administered to humans and other animals for treatment by any suitable route of administration, including oral, nasal (e.g., in spray form), rectal, intravaginal, parenteral, intracisternal, and topical (in powder, ointment, or drop form) administration, including buccal and sublingual administration.
Regardless of the route of administration chosen, the compounds of the invention and/or the pharmaceutical compositions of the invention, which may be used in a suitable hydrated form, are formulated into pharmaceutically acceptable dosage forms using conventional methods known to those skilled in the art.
The actual dosage level of the active ingredient in the pharmaceutical compositions of the invention can be varied to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition and mode of administration, and which is non-toxic to the patient.
The selected dosage level will depend upon a variety of factors including the activity of the particular compound of the invention or ester, salt or amide thereof employed, the route of administration, the time of administration, the rate of excretion of the particular compound employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound employed, the age, sex, body weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.
A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, a physician or veterinarian can start a dose of a compound of the invention employed in a pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect and gradually increase its dose until the desired effect is achieved.
In general, a suitable daily dose of a compound of the invention will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such effective dosages will generally depend on the factors recited above. In general, when used for the analgesic effects indicated, the compounds of the invention are administered to a patient in intravenous and subcutaneous doses of from about 0.0001 to about 100mg/kg body weight/day, more preferably from about 0.01 to about 50 mg/kg/day, and even more preferably from about 1.0 to about 100 mg/kg/day. An effective amount is an amount that treats a condition associated with a protein kinase.
If desired, an effective daily dose of the active compound may be administered at suitable intervals throughout the day in two, three, four, five, six or more sub-doses administered separately, optionally in unit dosage form.
For an individual of about 50-70kg, the pharmaceutical composition or combination of the invention may be in a unit dose of about 1-1000mg of the active ingredient, or about 1-500mg or about 1-250mg or about 1-150mg or about 0.5-100mg or about 1-50mg of the active ingredient. The therapeutically effective dose of the compound, pharmaceutical composition or combination thereof will depend on the species, weight, age and condition of the individual, the disorder or disease being treated or its severity. A physician, clinician or veterinarian of ordinary skill can readily determine the effective amount of each active ingredient which is required to prevent, treat or inhibit the progression of the disorder or disease.
The above dosage properties are illustrated for mammals, such as mice, rats, dogs, monkeys or their related organs, tissues or preparations, which are advantageous for use in vitro and in vivo assays. The compounds of the invention can be applied in vitro in the form of solutions, for example aqueous solutions, and in vivo enterally, parenterally, advantageously intravenously, for example as suspensions or aqueous solutions. The in vitro dosage may range between about 10-3 molar and 10-9 molar. The therapeutically effective amount in vivo can range from about 0.1 to 500mg/kg or between about 1 to 100mg/kg, depending on the route of administration.
The term "individual" as used herein means an animal. Typically, the animal is a mammal. A subject also means, for example, a primate (e.g., human, male or female), cow, sheep, goat, horse, dog, cat, rabbit, rat, mouse, fish, bird, and the like. In certain embodiments, the subject is a primate. In other embodiments, the individual is a human.
Although the compounds of the present invention may be administered alone, it is preferred to administer the compounds in the form of a pharmaceutical composition.
Pharmaceutical combination
Combination therapy using one or more compounds or compositions provided herein, or pharmaceutically acceptable derivatives thereof, in combination with other pharmaceutically active agents, is useful for treating the diseases and conditions described herein.
An effective amount of a compound or composition comprising a therapeutically effective concentration of a compound formulated for oral, systemic delivery, including parenteral or intravenous delivery or for topical or topical administration, is administered to an individual in need of treatment exhibiting symptoms of a disease or disorder. The amount is effective to treat, control or alleviate one or more symptoms of the disease or disorder.
One of ordinary skill in the art will appreciate that the compounds, isomers, prodrugs and pharmaceutically acceptable derivatives provided herein, including pharmaceutical compositions and formulations comprising such compounds, have broad application in combination therapy to treat the disorders and diseases described herein. Thus, the present invention contemplates the use of the compounds, isomers, prodrugs and pharmaceutically acceptable derivatives provided herein in combination with other active agents for the treatment of the diseases/disorders described herein.
General synthetic methods
In general, the compounds of the present invention may be prepared by the methods described herein, wherein the substituents are as defined for the compounds of formulae (I) - (II), unless otherwise specified. The following reaction schemes and examples serve to further illustrate the context of the invention.
Those skilled in the art will recognize that: the chemical reactions described herein may be used to suitably prepare a number of other compounds of the invention, and other methods for preparing the compounds of the invention are considered to be within the scope of the invention. For example, the synthesis of those non-exemplified compounds according to the present invention can be successfully accomplished by those skilled in the art by modification, such as appropriate protection of interfering groups, by the use of other known reagents in addition to those described herein, or by some routine modification of reaction conditions. In addition, the reactions disclosed herein or known reaction conditions are also recognized as being applicable to the preparation of other compounds of the present invention.
The examples described below, unless otherwise indicated, all temperatures were set to ℃. Reagents were purchased from commercial suppliers such as Aldrich Chemical Company, arco Chemical Company and Alfa Chemical Company and were used without further purification unless otherwise indicated. General reagents were purchased from Shantou Wen Long chemical reagent factory, guangdong Guanghua chemical reagent factory, guangzhou chemical reagent factory, tianjin HaoLiyu Chemicals Co., ltd, qingdao Tenglong chemical reagent Co., ltd, and Qingdao Kaseiki chemical plant.
The anhydrous tetrahydrofuran, dioxane, toluene and ether are obtained through reflux drying of metal sodium. Anhydrous DCM and chloroform were obtained by drying over calcium hydride under reflux. Ethyl acetate, petroleum ether, N-hexane, N, N-dimethylacetamide and N, N-dimethylformamide were used by previously drying over anhydrous sodium sulfate.
The following reactions are generally carried out under positive pressure of nitrogen or argon or by sleeving a dry tube over an anhydrous solvent (unless otherwise indicated), the reaction vial being stoppered with a suitable rubber stopper and the substrate being injected by syringe. The glassware was dried.
The column chromatography is performed using a silica gel column. Silica gel (300-400 mesh) was purchased from Qingdao oceanic plants. Nuclear magnetic resonance spectroscopy with CDC1 3 ，d 6 -DMSO，CD 3 OD or d 6 Acetone as solvent (reported in ppm) and TMS (0 ppm) or chloroform (7.25 ppm) as reference standard. When multiple peaks occur, the following abbreviations will be used: s (singleton), d (doublet), t (triplet ), m (multiplet, multiplet), br (broadenerd, broad), dd (doublet of doublets, quartet), dt (doublet of triplets, doublet). Coupling constants are expressed in hertz (Hz).
Low resolution Mass Spectral (MS) data were determined by Agilent 6320 series LC-MS spectrometer equipped with a G1312A binary pump and a G1316A TCC (column temperature maintained at 30 ℃), a G1329A autosampler and G1315B DAD detector applied for analysis, and an ESI source applied to the LC-MS spectrometer.
Low resolution Mass Spectral (MS) data were measured by Agilent 6120 series LC-MS spectrometer equipped with a G1311A quaternary pump and a G1316A TCC (column temperature maintained at 30 ℃), a G1329A autosampler and a G1315D DAD detector were used for analysis, and an ESI source was used for the LC-MS spectrometer.
Both spectrometers were equipped with an Agilent Zorbax SB-C18 column, 2.1X 30mm,5 μm in size. The injection volume is determined by the sample concentration; the flow rate is 0.6mL/min; peaks of HPLC were recorded by UV-Vis wavelength at 210nm and 254 nm. The mobile phases were 0.1% formic acid in acetonitrile (phase a) and 0.1% formic acid in ultrapure water (phase B). Gradient elution conditions are shown in table 1:
TABLE 1
The compound purification was assessed by Agilent 1100 series High Performance Liquid Chromatography (HPLC) with UV detection at 210nm and 254nm on a Zorbax SB-C18 column, 2.1X 30mm,4 μm,10 minutes, flow rate 0.6mL/min,5-95% (0.1% formic acid in acetonitrile) in (0.1% formic acid in water), the column temperature was maintained at 40 ℃.
The following acronyms are used throughout the invention:
BOC, boc tert-butoxycarbonyl
(pin) 2 B 2 Biboric acid pinacol ester
CHCl 3 Chloroform
CDCl 3 Deuterated chloroform
DCM dichloromethane
DMF N, N-dimethylformamide
DMSO dimethyl sulfoxide
d 6 -DMSO deuterated dimethyl sulfoxide
DIAD diisopropyl azodicarboxylate
EDC, EDCI 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride
HATU 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate
H 2 Hydrogen gas
MeOH，CH 3 OH methanol
mL, mL
N 2 Nitrogen gas
Pd/C Palladium/carbon
Pd(OAc) 2 Palladium acetate
Pd 2 (dba) 3 Tris (dibenzylideneacetone) dipalladium
PdCl 2 (dppf) 1,1' -bis-diphenylphosphinoferrocene palladium dichloride
AcOK Potassium acetate
EA ethyl acetate
XPhos-Pd-G2 chloro (2-dicyclohexylphosphino-2 ',4',6 '-triisopropyl-1,1' -biphenyl) [2- (2 '-amino-1,1' -biphenyl) ] palladium (II)
PPh 3 Triphenylphosphine
PE Petroleum Ether
HEX n-hexane
RT RT Room temperature
Rt Retention time
TFA trifluoroacetic acid
NBS N-bromosuccinimide
Cs 2 CO 3 Cesium carbonate
Xantphos 9,9-dimethyl-4,5-bisdiphenylphosphine xanthene
Synthetic schemes
The following examples are intended to further illustrate the present invention, but should not be construed as limiting the scope thereof.
Synthetic intermediates scheme 1
The compound intermediate (3 a) of the present invention can be obtained by the synthetic method of synthetic intermediate scheme 1: the compound (1 a) and the compound (2 a) are reacted under basic conditions by heating to produce a compound (3 a). Wherein Z, q and R 6 Have the meaning as described in the present invention.
Synthetic intermediates scheme 2
The compound of the present invention intermediate (3 b) can be obtained by the synthetic method of intermediate scheme 2: compounds (1 b) and (pin) 2 B 2 Under the condition of alkali, compound (2 b) is generated by reaction; the compound (2 b) is reacted with the compound (3 a) under basic conditions to obtain the compound (3 b). It is composed ofMiddle Y, Z, q and R 6 Have the meaning as described in the present invention.
Synthetic intermediates scheme 3
Intermediate (3 c) of the compounds of the present invention can be obtained by the synthetic method of synthetic intermediate scheme 3: the compound (3 b) and the compound (1 c) are reacted by heating to produce a compound (3 c). Wherein X, Y, Z, q and R 6 Have the meaning as described in the present invention.
Synthetic intermediates scheme 4
Intermediate (3 d) of the compounds of the present invention can be obtained by the synthetic method of intermediate scheme 4: reacting the compound (3 c) with the compound (1 d) under basic conditions to generate a compound (2 d); the compound (2 d) is reacted with (2 da) under basic conditions to give the compound (3 d). Wherein X, Y, Z, q, R 1 、R v And R 6 Have the meaning as described in the present invention.
Synthetic schemes
The compounds of the invention can be obtained by synthetic methods of the synthetic schemes: and (3) carrying out reduction reaction on the compound (3 d) and the compound (1 e) in a proper solvent to obtain a target product. Wherein X, Y, Z, q, R 1 、R 3 、R v And R 6 Have the meaning as described in the present invention.
The following examples may further illustrate the present invention, however, these examples should not be construed as limiting the scope of the present invention.
Examples
Example 1 (S) -1- (5-chloro-4- ((3 ' - (3- (2-cyano-7-azaspiro [3.5] nonan-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 1)
Step 1) [ 2-methyl-3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl ] methanol
PdCl 2 (dppf) (7.28g, 9.95mmol), acOK (29.3g, 299mmol) and pinacolate diboron (37.9g, 149mmol) were dissolved in 1,4-dioxane (200.1 mL) and (3-bromo-2-methyl-phenyl) methanol (20.1g, 100mmol) was added. The reaction mixture was warmed to 90 ℃ for 10h. The reaction mixture was stopped stirring and cooled to room temperature, then concentrated under reduced pressure. The resulting residue was isolated and purified by silica gel column chromatography (HEX/EA =10/1,v/v) to give the title compound as a pale green solid 23.9g with a yield of 96%;
LC-MS：(pos.ion)m/z：231.1[M-18+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ7.50(d，J＝7.3Hz，1H)，7.45(d，J＝7.4Hz，1H)，7.14(t，J＝7.5Hz，1H)，5.05(t，J＝5.4Hz，1H)，4.48(d，J＝5.3Hz，2H)，2.39(s，3H)，1.29(s，12H).
step 2) 1-bromo-3- (3-bromopropoxy) -2-methylbenzene
3-bromo-2-methylphenol (15.1g, 80.7 mmol) was dissolved in acetone (180.1 mL), potassium carbonate (33.3g, 241mmol) was added, 1,3-dibromopropane (20.4mL, 201mmol) was added, nitrogen was used for protection, and the reaction mixture was heated under reflux for 12h. Stirring was stopped, cooled to room temperature, filtered with suction, and the filter cake was washed with DCM, then the filtrate was concentrated under reduced pressure. The resulting concentrated residue was separated and purified by silica gel column chromatography (n-hexane) to give the title compound as a colorless oil 24.1g, in 97% yield;
1 H NMR(400MHz，CDCl 3 )δ7.20(d，J＝8.0Hz，1H)，7.03(t，J＝8.1Hz，1H)，6.82(d，J＝8.2Hz，1H)，4.12(t，J＝5.7Hz，2H)，3.65(t，J＝6.4Hz，2H)，2.40-2.36(m，2H)，2.35(s，3H).
step 3) (3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methanol
[ 2-methyl-3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl ] methanol (20.0g, 80.6 mmol) and 1-bromo-3- (3-bromopropoxy) -2-methylbenzene (26.1g, 84.7 mmol) were dissolved in THF (400.1 mL), and a potassium phosphate solution (400.1mL, 200mmol) was added. The reaction mixture was stirred at room temperature under nitrogen for 20min, then XPhos-Pd-G2 (1.27g, 1.62mmol) was added and stirring at room temperature was continued for 10h under nitrogen. After completion of the reaction, the mixture was diluted with water (300 mL), extracted with ethyl acetate (150 mL. Times.3), and the organic phases were combined. The combined organic phases were dried over anhydrous sodium sulfate and then concentrated under reduced pressure, and the residue was isolated and purified by silica gel column chromatography (PE/EA =3/1,v/v) to give the title compound 15.9g of orange-red viscous oil in 57% yield;
LC-MS：(pos.ion)m/z：332.0[M-18+1] + ；
1 H NMR(400MHz，CDCl 3 )δ7.41(d，J＝7.5Hz，1H)，7.27(t，J＝7.3Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.11(d，J＝7.4Hz，1H)，6.90(d，J＝8.1Hz，1H)，6.78(d，J＝7.5Hz，1H)，4.78(s，2H)，4.27-4.09(m，2H)，3.68(t，J＝6.5Hz，2H)，2.45-2.36(m，2H)，2.08(s，3H)，1.95(s，3H).
step 4) 4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
(3 ' - (3-Bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methanol (15.9 g,45.5 mmol), 5-chloro-2,4-dihydroxy-benzaldehyde (8.64g, 50.1 mmol) were dissolved in THF (242.1 mL) at 0 deg.C, PPh3 (17.9 g,68.2 mmol) was added, nitrogen blanketed, DIAD (13.4 mL, 68.1mmol) was slowly injected, and stirred at room temperature for 5h. After the reaction was completed, the stirring was stopped, and water (100 mL) was added for dilution, and extraction was performed with ethyl acetate (100 mL. Times.3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Separating and purifying by silica gel column chromatography (PE/EA =8/1,v/v) to obtain 5.3g of yellow solid with the yield of 23%;
LC-MS：(pos.ion)m/z：504.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ11.16(s，1H)，10.03(s，1H)，7.71(s，1H)，7.50(d，J＝7.4Hz，1H)，7.29(t，J＝7.6Hz，1H)，7.22(t，J＝7.9Hz，1H)，7.09(d，J＝7.4Hz，1H)，6.98(d，J＝8.2Hz，1H)，6.87(s，1H)，6.70(d，J＝7.5Hz，1H)，5.31(s，2H)，4.18-4.04(m，2H)，3.71(t，J＝6.5Hz，2H)，2.29(dd，J＝12.3，6.1Hz，2H)，2.01(s，3H)，1.84(s，3H).
step 5) 7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-azaspiro [3.5] nonane-2-carbonitrile
4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (1 g, 1.98mmol) and 7-azaspiro [3.5] nonane-2-carbonitrile (0.357g, 2.37mmol) were dissolved in DMF (15 mL), potassium carbonate (0.6857 g, 4.96mmol) was added, nal (0.357g, 2.38mmol) was added, nitrogen was added and the mixture was stirred at 75 ℃ for 13h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography purification (DCM/MeOH =10/1,v/v) gave 0.71g of light yellow solid in 62.4% yield;
LC-MS：(pos.ion)m/z：573.2[M+1] + 。
step 6) 7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-aza-spiro [3.5] nonane-2-carbonitrile
5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (0.295g, 1.48mmol) was dissolved in DMF (12 mL), and cesium carbonate (1g, 3.1mmol) was added and stirred at room temperature for 10min. 7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) endo) -7-azaspiro [3.5] nonane-2-carbonitrile (0.71g, 1.24mmol) and NaI (18mg, 0.124mmol) were added, with nitrogen protection, and the mixture was stirred at 75 ℃ for 4h. Stirring was then discontinued, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 160mg of a yellow solid in 18.74% yield;
LC-MS：(pos.ion)m/z：689.2[M+1] + 。
step 7) (S) -1- (5-chloro-4- ((3 ' - (3- (2-cyano-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-aza-spiro [3.5] nonane-2-carbonitrile (160mg, 0.23mmol) and D-pipecolic acid (0.044g, 0.34mmol) were dissolved in DMF (15.0 mL), adjusted to pH 5 by the addition of acetic acid, heated to 60 ℃ and stirred for 1h, then cooled to room temperature, sodium cyanoborohydride (0.072g, 1.15mmol) was slowly added, protected with nitrogen, stirred for 12h at room temperature. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. Silica gel column chromatography (DCM/MeOH =8/1,v/v) gave 47mg yellow solid isolated in 25.23% yield;
LC-MS：(pos.ion)m/z：802.3[M+1] + ；
1 H NMR(600MHz，d 6 -DMSO)δ9.00(d，J＝10.7Hz，2H)，8.46(s，1H)，7.48(d，J＝7.4Hz，1H)，7.41(s，1H)，7.26(t，J＝7.5Hz，1H)，7.20(t，J＝7.8Hz，1H)，7.11(s，1H)，7.06(d，J＝7.5Hz，1H)，6.94(d，J＝8.2Hz，1H)，6.67(d，J＝7.4Hz，1H)，5.36-5.29(m，2H)，5.29-5.23(m，2H)，4.02(dt，J＝22.1，7.6Hz，2H)，3.77(d，J＝13.8Hz，1H)，3.60(d，J＝13.6Hz，1H)，3.51(s，1H)，3.31(dt，J＝17.1，8.6Hz，2H)，3.13(s，1H)，2.88(d，J＝6.7Hz，1H)，2.38(s，2H)，2.28(d，J＝5.7Hz，2H)，2.16(t，J＝10.2Hz，2H)，2.02(s，3H)，2.00(d，J＝11.7Hz，2H)，1.92(d，J＝6.2Hz，2H)，1.91(s，1H)，1.81(s，3H)，1.79-1.76(m，1H)，1.72(d，J＝8.9Hz，1H)，1.62(s，2H)，1.58(s，2H)，1.48(s，3H)，1.37(s，1H).
example 2 (S) -1- (4- ((3 ' - (3- (3-oxa-9-azaspiro [5.5] undec-9-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 2)
Step 1) 4- ((3 ' - (3- (3-oxa-9-azaspiro [5.5] undec-9-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) -methoxy) -5-chloro-2-hydroxybenzaldehyde
4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) -methoxy) -5-chloro-2-hydroxy-benzaldehyde (0.65g, 1.29mmol) and 9-oxa-3-azaspiro [5.5] undecane (0.24g, 1.54mmol) were dissolved in DMF (15 mL), potassium carbonate (0.445g, 3.22mmol) was added, naI (0.23g, 1.55mmol) was added, nitrogen was added and the mixture was stirred for 13h while warming to 75 ℃. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Column chromatography on silica gel (DCM/MeOH =8/1,v/v) gave 0.51g of a light yellow solid in 68.3% yield;
LC-MS：(pos.ion)m/z：578.2[M+1] + 。
step 2) 5- ((5- ((3 ' - (3- (3-oxa-9-azaspiro [5.5] undec-9-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (0.161g, 1.05mmol) was dissolved in DMF (12 mL), cesium carbonate (0.574g, 1.76mmol) was added, and stirring was carried out at room temperature for 10min. 4- ((3 ' - (3- (3-oxa-9-azaspiro [5.5] undec-9-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (0.51g, 0.88mmol) and NaI (13mg, 0.088mmol) were added, with nitrogen protection, and the temperature was raised to 75 ℃ and stirred for 4h. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 175mg of yellow solid in 28.5% yield;
LC-MS：(pos.ion)m/z：694.3[M+1] + 。
step 3) (S) -1- (4- ((3 ' - (3- (3-oxa-9-azaspiro [5.5] undec-9-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
5- ((5- ((3 ' - (3- (3-oxa-9-azaspiro [5.5] undec-9-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (0.17g, 0.244mmol) and D-pipecolic acid (0.063g, 0.487mmol) were dissolved in DMF (15.0 mL), acetic acid was added to adjust the pH to around 5, heated to 60 ℃ and stirred for 1h, cooled to room temperature, sodium cyanoborohydride (0.07g, 1.21mmol) was slowly added, protected with nitrogen, and stirred for 12h at room temperature. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. Separating and purifying by silica gel column chromatography (DCM/MeOH =8/1,v/v), and separating to obtain 38mg of brown yellow solid with the yield of 19.2%;
LC-MS：(pos.ion)m/z：807.4[M+1] + ；
1 H NMR(600MHz，d 6 -DMSO)δ9.00(d，J＝9.3Hz，2H)，8.46(s，1H)，7.49(d，J＝7.3Hz，1H)，7.41(s，1H)，7.26(t，J＝7.4Hz，1H)，7.20(t，J＝7.7Hz，1H)，7.11(s，1H)，7.06(d，J＝7.4Hz，1H)，6.95(d，J＝8.1Hz，1H)，6.68(d，J＝7.4Hz，1H)，5.34(d，J＝13.5Hz，2H)，5.27(d，J＝12.7Hz，2H)，4.08-3.99(m，2H)，3.77(d，J＝13.7Hz，1H)，3.61(d，J＝13.7Hz，1H)，3.52(d，J＝7.6Hz，6H)，3.14(s，1H)，2.89(s，1H)，2.67(s，2H)，2.58(s，3H)，2.28(s，1H)，2.02(s，3H)，1.99(s，2H)，1.81(s，3H)，1.80-1.76(m，1H)，1.72(s，1H)，1.55(s，3H)，1.48(s，3H)，1.40(s，5H).
example 3 (S) -1- (4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 3)
Step 1) 2-oxa-8-azaspiro [4.5] decane
Under the protection of nitrogen, dissolving 2-oxa-8-azaspiro [4.5] decane-8-carboxylic acid tert-butyl ester in trifluoroacetic acid (4 mL) and DCM (25 mL), reacting at room temperature for 23h, and concentrating the reaction solution to obtain a tan oily liquid 585.2mg with a yield of 51%;
LC-MS：(pos.ion)m/z：142.1[M+1] + 。
step 2) 4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
2-oxa-8-azaspiro [4.5]]Decane (300mg, 2.1245mmol) and 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl]-2-methyl-phenyl]Methoxy radical]-5-chloro-2-hydroxy-benzaldehyde (1.04g, 2.06mmol) dissolved in DMF (15 mL) and K added sequentially 2 CO 3 (600mg, 4.34mmol) and NaI (300mg, 2.00mmol) were heated to 75 ℃ and reacted for 17h. Extracted with ethyl acetate (30 mL. Times.3), washed with water (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Separation and purification by silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 350mg of an orange-red solid with a yield of 30%;
LC-MS：(pos.ion)m/z：564.1[M+1] + 。
step 3) 4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridine-3-methoxy) benzaldehyde
4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzene (350mg, 0.62mmol) and 5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (123mg, 0.80mmol) were dissolved in N, N-dimethylformamide (20 mL), cesium carbonate (404mg, 1.23mmol) and sodium iodide (9.3mg, 0.06mmol) were added, heated to 75 ℃ and reacted for 3h. The reaction was cooled to room temperature, extracted with ethyl acetate (30 mL. Times.3), washed with water (50 mL), dried over anhydrous sodium sulfate, and filtered. The concentrated crude product was purified by silica gel column chromatography (DCM/MeOH =10/1,v/v) to give 342mg of a yellow solid in 81% yield;
LC-MS：(pos.ion)m/z：681.4[M+1] + 。
step 4) (S) -1- (4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridine-3-methoxy) benzaldehyde (342mg, 0.50mmol) and (2S) -piperidine-2-carboxylic acid (130mg, 1.00mmol) were dissolved in N, N-dimethylformamide (10 mL), acetic acid (0.1mL, 2.0mmol) was added, and the mixture was heated to 60 ℃ for 1.5 hours. After cooling to room temperature, sodium cyanoborohydride (158mg, 2.51mmol) was slowly added and reacted at room temperature for 16 hours. Subsequently, the reaction mixture was heated to 80 ℃ and reacted for 5 hours. The reaction was cooled to room temperature, and a saturated potassium carbonate solution (30 mL) was added and stirred at room temperature for 30min. The reaction was stopped, extracted with ethyl acetate (30 mL. Times.3), washed with water (50 mL), dried over anhydrous sodium sulfate, and filtered. The concentrated crude product was purified by silica gel column chromatography (DCM/MeOH =10/1,v/v) to give 60mg of a pale yellow solid in 15% yield;
LC-MS：(pos.ion)m/z：794.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.00(d，J＝5.2Hz，2H)，8.46(s，1H)，7.49(d，J＝7.3Hz，1H)，7.41(s，1H)，7.27(t，J＝7.6Hz，1H)，7.21(d，J＝7.7Hz，1H)，7.12(s，1H)，7.06(d，J＝7.5Hz，1H)，6.97(d，J＝8.3Hz，1H)，6.70(d，J＝7.4Hz，1H)，5.33(s，2H)，5.27(s，2H)，4.08(d，J＝7.0Hz，2H)，3.74(t，J＝7.1Hz，3H)，3.62(d，J＝13.5Hz，2H)，3.51(s，2H)，3.15(s，2H)，2.95(s，3H)，2.89(s，3H)，2.29(s，1H)，2.13(s，2H)，2.03(s，3H)，1.83(s，3H)，1.78(s，1H)，1.71(d，J＝7.2Hz，6H)，1.48(s，3H)，1.38(s，1H).
example 4 (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,7-diazaspiro [4.4] non-7-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 4)
Step 1) 5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,7-diazaspiro [4.4] non-7-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde
Reacting 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl]-2-methyl-phenyl]Methoxy radical]-5-chloro-2-hydroxy-benzaldehyde (1.5g, 3.0mmol) and 4,8-diazaspiro [4.4]]Nonan-3-one (500mg, 3.56mmol) was dissolved in N, N-dimethylformamide (25 mL), and K was added sequentially 2 CO 3 (1g, 7.23mmol) and NaI (540 mg, 3.60mmol), heated to 75 ℃ and reacted for 23h. The reaction was cooled to room temperature, extracted with ethyl acetate (30 mL. Times.3), washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The concentrated crude product was isolated and purified by silica gel column chromatography (DCM/MeOH =10/1,v/v). 365mg of orange-red solid powder is obtained, and the yield is 22%;
LC-MS：(pos.ion)m/z：564.4[M+1] + 。
step 2) 5- ((4-chloro-5- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,7-diazaspiro [4.4] non-7-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-formylphenoxy) methyl) nicotinonitrile
5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,7-diazaspiro [4.4] non-7-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde (365mg, 0.64mmol) and 5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (160mg, 0.84mmol) were dissolved in N, N-dimethylformamide (25 mL), cesium carbonate (634mg, 1.94mmol) and sodium iodide (9.7mg, 0.065mmol) were added, heated to 75 ℃ and reacted for 3 hours. After the reaction was cooled to room temperature, it was extracted with ethyl acetate (30 mL. Times.3), washed with water (50 mL), dried over anhydrous sodium sulfate, and filtered. The concentrated crude product was purified by silica gel column chromatography (DCM/MeOH =10/1,v/v) to give 100mg of yellow solid in 81% yield;
LC-MS：(pos.ion)m/z：601.1[M+1] + 。
step 3) (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,7-diazaspiro [4.4] non-7-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
5- ((4-chloro-5- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,7-diazaspiro [4.4] non-7-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-formylphenoxy) methyl) nicotinonitrile (100mg, 0.15mmol) and (2S) -piperidine-2-carboxylic acid (38mg, 0.29mmol) were dissolved in N, N-dimethylformamide (10 mL), acetic acid (0.1 mL) was added, heated to 60 ℃ and stirred for 1.5h. The heating was stopped, cooled to room temperature, and sodium cyanoborohydride (46.3 mg, 0.74mmol) was added and reacted at room temperature for 14h. The reaction was heated to 80 ℃ and reacted for 5h. The reaction was cooled to room temperature, and saturated potassium carbonate solution (30 mL) was added and stirred for 30 minutes. The reaction was stopped, extracted with ethyl acetate (30 mL. Times.3), washed with water (50 mL), dried over anhydrous sodium sulfate, and filtered. The concentrated crude product was purified by silica gel column chromatography (DCM/MeOH =10/1,v/v) to give 28mg of a pale red solid in 24% yield;
LC-MS：(pos.ion)m/z：793.5[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(s，2H)，8.47(s，1H)，7.93(s，1H)，7.49(d，J＝7.4Hz，1H)，7.43(s，1H)，7.27(t，J＝7.7Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.14(s，1H)，7.07(d，J＝7.3Hz，1H)，6.96(d，J＝8.2Hz，1H)，6.68(d，J＝7.4Hz，1H)，5.31(d，J＝30.5Hz，4H)，4.07(d，J＝5.9Hz，2H)，3.80(d，J＝14.0Hz，1H)，3.64(d，J＝13.6Hz，2H)，3.48(d，J＝5.2Hz，1H)，3.42(d，J＝4.6Hz，1H)，3.17(s，2H)，2.88(s，1H)，2.67(s，4H)，2.31(s，1H)，2.17(dd，J＝15.5，7.3Hz，2H)，2.03(s，3H)，1.96(s，2H)，1.91(s，2H)，1.82(s，3H)，1.81-1.76(m，1H)，1.73(s，1H)，1.49(s，3H)，1.37(s，1H).
example 5 (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (6-hydroxy-2-azaspiro [3.4] oct-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 5)
Step 1) 5-chloro-2-hydroxy-4- ((3 ' - (3- (6-hydroxy-2-azaspiro [3.4] oct-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzaldehyde
4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (0.8g, 1.58mmol) and 2-aza-spiro [3.4] oct-7-ol (0.242g, 1.58mmol) were dissolved in DMF (15 mL), potassium carbonate (0.658g, 4.76mmol) was added, naI (0.285g, 1.90mmol) was added, nitrogen was added and the mixture was stirred at 75 ℃ for 13h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 0.37g of light yellow viscous, 42.3% yield;
LC-MS：(pos.ion)m/z：550.2[M+1] + 。
step 2) 5- ((4-chloro-2-formyl-5- ((3 ' - (3- (6-hydroxy-2-azaspiro [3.4] oct-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile
5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (0.17g, 0.85mmol) was dissolved in DMF (12 mL), cesium carbonate (0.533g, 1.64mmol) was added, and the mixture was stirred at room temperature for 10min. 5-chloro-2-hydroxy-4- ((3 ' - (3- (6-hydroxy-2-azaspiro [3.4] oct-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzaldehyde (360mg, 0.65mmol) and NaI (9mg, 0.065 mmol) were added, and the system was stirred at 75 ℃ for 4h under nitrogen protection. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 0.135g of a yellow solid in 30.96% yield;
LC-MS：(pos.ion)m/z：666.2[M+1] + 。
step 3) (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (6-hydroxy-2-azaspiro [3.4] oct-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
5- ((4-chloro-2-formyl-5- ((3 ' - (3- (6-hydroxy-2-aza-spiro [3.4] oct-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile (134mg, 0.2011mmol) and D-pipecolic acid (0.038g, 0.29mmol) were dissolved in DMF (15.0 mL), acetic acid was added to adjust the pH to about 5, heated to 60 ℃ and stirred for 1h, cooled to room temperature, sodium cyanoborohydride (0.063g, 1.00mmol) was slowly added, nitrogen protected, and stirred for 12h at room temperature. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. Separating and purifying by silica gel column chromatography (DCM/MeOH =8/1,v/v), and separating to obtain 0.025g of yellow solid with the yield of 15.9%;
LC-MS：(pos.ion)m/z：779.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.00(d，J＝2.8Hz，2H)，8.46(s，1H)，7.49(d，J＝7.5Hz，1H)，7.42(s，1H)，7.27(t，J＝7.5Hz，1H)，7.21(t，J＝7.8Hz，1H)，7.13(s，1H)，7.06(d，J＝7.5Hz，1H)，6.96(d，J＝8.2Hz，1H)，6.70(d，J＝7.5Hz，1H)，5.34(s，2H)，5.27(s，2H)，4.08(s，2H)，3.79(d，J＝13.4Hz，2H)，3.63(d，J＝14.0Hz，2H)，3.51(s，2H)，3.16(s，2H)，2.89(s，2H)，2.30(s，2H)，2.03(s，4H)，1.96(s，2H)，1.89(d，J＝4.4Hz，1H)，1.83(d，J＝1.8Hz，3H)，1.79(s，1H)，1.76-1.66(m，3H)，1.48(s，5H)，1.37(s，2H).
example 159 (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (hexahydro-2H-pyrano [3,2-c ] pyridin-6 (7H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 159)
Step 1) 5-chloro-4- ((3 ' - (3- (hexahydro-2H-pyrano [3,2-c ] pyridin-6 (7H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde
4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxybenzaldehyde (0.6g, 1.19mmol) and trans-3, 4,4a,5,6,7,8, 8a-octahydro-2H-pyrano [3,2-c ] pyridine (0.2g, 1.42mmol) were dissolved in DMF (15 mL), potassium carbonate (0.197g, 1.42mmol) was added, naI (0.214g, 1.42mmol) was added, and the system was stirred at 75 ℃ for 13H under nitrogen protection. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Purification by silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 0.38g of pale yellow thick substance in 56.65% yield;
LC-MS：(pos.ion)m/z：564.2[M+1] + 。
step 2) 5- ((4-chloro-2-formyl-5- ((3 '- (3- (hexahydro-2H-pyrano [3,2-c ] pyridin-6 (7H) -yl) propoxy) -2,2-dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile
5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (0.175g, 0.879mmol) was dissolved in DMF (12 mL), and cesium carbonate (0.548g, 1.68mmol) was added and stirred at room temperature for 10min. 5-chloro-4- ((3 ' - (3- (hexahydro-2H-pyrano [3,2-c ] pyridin-6 (7H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde (0.38mg, 0.67mmol) and NaI (10mg, 0.067mmol) were added, the mixture was warmed to 75 ℃ under nitrogen and stirred for 4H. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 0.128g of a yellow solid in 27.9% yield;
LC-MS：(pos.ion)m/z：680.3[M+1] + 。
step 3) (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (hexahydro-2H-pyrano [3,2-c ] pyridin-6 (7H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
5- ((4-chloro-2-formyl-5- ((3 '- (3- (hexahydro-2H-pyrano [3,2-c ] pyridin-6 (7H) -yl) propoxy) -2,2-dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile (0.123g, 0.180mmol) and D-pipecolic acid (0.035g, 0.27mmol) were dissolved in DMF (15.0 mL), adjusted to pH 5 by the addition of acetic acid, heated to 60 ℃ and stirred for 1H, cooled to room temperature, sodium cyanoborohydride (0.056g, 0.89mmol) was slowly added, with nitrogen protection, and stirred for 12H at room temperature. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added, stirred at room temperature for 30min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Silica gel column chromatography (DCM/MeOH =8/1,v/v) gave 0.02g of light yellow solid with 13.9% yield.
LC-MS：(pos.ion)m/z：793.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(d，J＝2.0Hz，2H)，8.47(s，1H)，7.49(d，J＝7.5Hz，1H)，7.42(s，1H)，7.27(t，J＝7.5Hz，1H)，7.22(t，J＝7.9Hz，1H)，7.13(s，1H)，7.07(d，J＝7.5Hz，1H)，6.97(d，J＝8.2Hz，1H)，6.70(d，J＝7.5Hz，1H)，5.38-5.30(m，2H)，5.29(d，J＝13.7Hz，2H)，4.09(d，J＝6.2Hz，2H)，3.90(d，J＝10.2Hz，1H)，3.82-3.75(m，2H)，3.66(s，1H)，3.62(s，1H)，3.58(s，2H)，3.41(s，2H)，3.16(s，2H)，2.89(s，1H)，2.31(d，J＝13.1Hz，1H)，2.24(s，2H)，2.16(s，1H)，2.03(s，4H)，1.84(s，4H)，1.79(s，1H)，1.74(d，J＝4.8Hz，2H)，1.60(d，J＝12.1Hz，2H)，1.49(s，3H)，1.38(s，1H)，1.29(d，J＝8.7Hz，2H).
Example 7 (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (hexahydrofuro [3,4-c ] pyridin-5 (3H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 7)
Step 1) 5-chloro-4- ((3 ' - (3- (hexahydrofuro [3,4-c ] pyridin-5 (3H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde
4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (0.6g, 1.19mmol) and 1, 3a,4,5,6,7 a-octahydrofuro [3,4-c ] pyridine hydrochloride (0.233mg, 1.42mmol) were dissolved in DMF (15 mL), potassium carbonate (0.411g, 2.97mmol) was added, naI (0.214g, 1.43mmol) was added, nitrogen was added and the mixture was stirred at 75 ℃ for 13h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography isolation and purification (DCM/MeOH =10/1,v/v) gave 0.4g of a light yellow solid in 61.06% yield.
LC-MS：(pos.ion)m/z：550.2[M+1] + ；
Step 2) 5- ((4-chloro-2-formyl-5- ((3 ' - (3- (hexahydrofuro [3,4-c ] pyridin-5 (3H) -yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile
5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (0.188g, 0.94mmol) was dissolved in DMF (12 mL), and cesium carbonate (0.592g, 1.82mmol) was added and stirred at room temperature for 10min. 5-chloro-4- ((3 ' - (3- (hexahydrofuro [3,4-c ] pyridin-5 (3H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde (400mg, 0.73mmol) and NaI (10mg, 0.073mmol) were added, stirred at 75 ℃ under nitrogen. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography (DCM/MeOH =8/1,v/v) gave 400mg of a yellow solid in 82.57% yield.
LC-MS：(pos.ion)m/z：666.2[M+1] + ；
Step 3) (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (hexahydrofuro [3,4-c ] pyridin-5 (3H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
5- ((4-chloro-2-formyl-5- ((3 ' - (3- (hexahydrofuro [3,4-c ] pyridin-5 (3H) -yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile (0.4 mg, 0.60mmol) and D-pipecolic acid (0.118g, 0.91mmol) were dissolved in DMF (15.0 mL), adjusted to pH 5 by the addition of acetic acid, heated to 60 ℃ and stirred for 1H, cooled to room temperature, sodium cyanoborohydride (0.192g, 3.05mmol) was slowly added, protected with nitrogen, and stirred at room temperature for 12H. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. Purification by silica gel column chromatography (DCM/MeOH =8/1,v/v) gave 70mg of a pale yellow solid isolated in 14.96% yield.
LC-MS：(pos.ion)m/z：779.5[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(s，2H)，8.46(s，1H)，7.49(d，J＝7.4Hz，1H)，7.42(s，1H)，7.26(t，J＝7.5Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.13(s，1H)，7.06(d，J＝7.5Hz，1H)，6.96(d，J＝8.2Hz，1H)，6.69(d，J＝7.5Hz，1H)，5.38-5.30(m，2H)，5.27(s，2H)，4.07(d，J＝6.4Hz，2H)，3.82(d，J＝5.7Hz，1H)，3.77(s，1H)，3.72(dd，J＝16.6，7.9Hz，3H)，3.65(s，1H)，3.60(d，J＝16.9Hz，3H)，3.16-3.13(m，1H)，2.88(s，2H)，2.30(s，2H)，2.11(s，2H)，2.03(s，4H)，1.83(s，5H)，1.72(d，J＝9.7Hz，3H)，1.49(s，3H)，1.37(s，2H).
Example 8 (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (hexahydro-1H-pyrrolo [3,4-c ] pyridin-5 (6H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 8)
Step 1) tert-butyl 5- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) hexahydro-1H-pyrrolo [3,4-c ] pyridine-2 (3H) -carboxylate
4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (1g, 1.98mmol) and 1, 3a,4,5,6,7 a-octahydropyrrolo [3,4-c ] pyridine-2-carboxylic acid tert-butyl ester hydrochloride (0.625g, 2.37mmol) were dissolved in DMF (15 mL), potassium carbonate (0.685 g, 4.96mmol) was added, naI (0.357g, 2.38mmol) was added, nitrogen blanketed, warmed to 75 ℃ and stirred for 13h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Column chromatography on silica gel (DCM/MeOH =10/1,v/v) gave 1.03g of a light yellow viscous material in 79.9% yield.
LC-MS：(pos.ion)m/z：649.3[M+1] + ；
Step 2) tert-butyl 5- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) hexahydro-1H-pyrrolo [3,4-c ] pyridine-2 (3H) -carboxylate
5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (0.44g, 2.21mmol) was dissolved in DMF (12 mL), cesium carbonate (1.4 g, 4.2mmol) was added, and the mixture was stirred at room temperature for 10min. 5- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) hexahydro-1H-pyrrolo [3,4-c ] pyridine-2 (3H) -carboxylic acid tert-butyl ester (1.1g, 1.7 mmol) and NaI (0.025g, 0.17mmol) were added, with nitrogen blanket, warmed to 75 ℃ and stirred for 4H. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 0.482g of a yellow solid in 37% yield.
LC-MS：(pos.ion)m/z：765.3[M+1] + ；
Step 3) (2S) -1- (4- ((3 ' - (3- (2- (tert-butoxycarbonyl) hexahydro-1H-pyrrolo [3,4-c ] pyridin-5 (6H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
Tert-butyl 5- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) hexahydro-1H-pyrrolo [3,4-c ] pyridine-2 (3H) -carboxylate (0.482g, 0.63mmol) and D-pipecolic acid (0.122g, 0.94mmol) were dissolved in DMF (15.0 mL), adjusted to pH 5 by addition of acetic acid, heated to 60 ℃ and stirred for 1H, cooled to room temperature, sodium cyanoborohydride (0.1973g, 3.1mmol) was slowly added, protected with nitrogen, stirred for 12H at room temperature. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. Purification by silica gel column chromatography (DCM/MeOH =8/1,v/v) afforded 0.183g of a yellow solid isolated in 33.07% yield.
LC-MS：(pos.ion)m/z：878.4[M+1] + ；
Step 4) (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (hexahydro-1H-pyrrolo [3,4-c ] pyridin-5 (6H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
(2S) -1- (4- (3 ' - (3- (2- (tert-butoxycarbonyl) hexahydro-1H-pyrrolo [3,4-c ] pyridin-5 (6H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (0.183g, 0.20mmol) was dissolved in DCM (20 mL), and TFA (2 mL) was added and the reaction was stirred at room temperature for 5 hours. Stirring was stopped, the solvent was concentrated, a saturated potassium carbonate solution (40 mL) was added, the reaction was stirred at room temperature for 15min, extracted with ethyl acetate (30 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by thin layer chromatography (DCM/MeOH = 6/1) gave 0.043g of white solid in 26.5% yield.
LC-MS：(pos.ion)m/z：778.3[M+1] + ；
1 H NMR(600MHz，d 6 -DMSO)δ9.62(s，1H)，9.04-8.99(m，2H)，8.48(s，1H)，7.50(d，J＝7.5Hz，1H)，7.43(s，1H)，7.27(t，J＝7.5Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.15(s，1H)，7.07(d，J＝7.3Hz，1H)，6.96(d，J＝8.3Hz，1H)，6.68(d，J＝7.5Hz，1H)，5.40-5.32(m，2H)，5.31-5.24(m，2H)，4.04(dd，J＝11.9，6.2Hz，2H)，3.81(d，J＝13.6Hz，1H)，3.66(d，J＝13.6Hz，1H)，3.51(s，1H)，3.21-3.12(m，5H)，2.98(d，J＝8.7Hz，1H)，2.91(s，1H)，2.62(s，2H)，2.33(s，3H)，2.22(s，1H)，2.10(s，1H)，2.03(s，3H)，1.92(s，2H)，1.82(d，J＝2.1Hz，4H)，1.72(d，J＝9.6Hz，1H)，1.63(s，1H)，1.57(s，1H)，1.50(s，3H)，1.42-1.32(m，2H).
Example 9 (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 9)
Step 1) 5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde
4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (0.9 g, 1.78mmol), 3a,4,5,6 a-hexahydro-1H-furo [3,4-c ] pyrrole hydrochloride (0.32g, 2.14mmol) were dissolved in DMF (15 mL), potassium carbonate (0.62g, 4.46mmol) was added, naI (0.32g, 2.14mmol) was added, nitrogen was added and the mixture was stirred at 75 ℃ for 13 hours. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Column chromatography on silica gel (DCM/MeOH =8/1,v/v) gave 0.62g of a light yellow solid in 64.7% yield.
LC-MS：(pos.ion)m/z：536.2[M+1] + ；
Step 2) 5- ((4-chloro-5- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-formylphenoxy) methyl) nicotinonitrile
5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (0.273g, 1.38mmol) was dissolved in DMF (12 mL), cesium carbonate (0.753g, 2.31mmol) was added, and the mixture was stirred at room temperature for 10min. 5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde (0.62g, 1.15mmol) and NaI (17mg, 0.115mmol) were added, warmed to 75 ℃ under nitrogen and stirred for 4H. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Purification by silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 112mg of yellow solid in 14.85% yield.
LC-MS：(pos.ion)m/z：652.2[M+1] + ；
Step 3) (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
5- ((4-chloro-5- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-formylphenoxy) methyl) nicotinonitrile (0.112g, 0.17mmol) and D-pipecolic acid (0.033g, 0.255mmol) were dissolved in DMF (15.0 mL), adjusted to pH 5 by adding acetic acid, heated to 60 ℃ and stirred for 1H, cooled to room temperature, sodium cyanoborohydride (0.053g, 0.84mmol) was slowly added, protected with nitrogen, stirred for 12H at room temperature. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. Purification by silica gel column chromatography (DCM/MeOH =8/1,v/v) afforded 22mg of a yellow solid isolated in 16.7% yield.
LC-MS：(pos.ion)m/z：765.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(s，2H)，8.46(s，1H)，7.49(d，J＝7.5Hz，1H)，7.42(s，1H)，7.26(t，J＝7.5Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.12(s，1H)，7.07(d，J＝7.3Hz，1H)，6.95(d，J＝8.3Hz，1H)，6.68(d，J＝7.5Hz，1H)，5.33(s，2H)，5.26(s，2H)，4.06(d，J＝6.2Hz，2H)，3.79(d，J＝13.6Hz，1H)，3.65(s，3H)，3.15(s，2H)，2.89(s，3H)，2.80(s，4H)，2.31(d，J＝6.8Hz，2H)，2.03(s，6H)，1.82(s，4H)，1.73(s，1H)，1.49(s，3H)，1.35(d，J＝15.1Hz，2H).
Example 10 (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (5-hydroxyhexahydro-dihydrocyclopenta [ c ] pyrrole-2 (1H) -propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 10)
Step 1) 5-chloro-2-hydroxy-4- ((3 ' - (3- (5-hydroxyhexadienocyclo [ c ] pyrrol-2 (1H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzaldehyde
4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (0.8g, 1.59mmol) and 1,2,3,4,5,6 a-octahydrocyclopenta [ c ] pyrrol-5-ol (0.242g, 1.90mmol) were dissolved in DMF (15 mL), potassium carbonate (0.658g, 4.76mmol) was added, naI (0.285g, 1.90mmol) was added, nitrogen was added and stirring was continued at 75 ℃ for 13h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Column chromatography on silica gel (DCM/MeOH =10/1,v/v) gave 0.7g of a light yellow viscous, 80.14% yield.
LC-MS：(pos.ion)m/z：550.2[M+1] + ；
Step 2) 5- ((4-chloro-2-formyl- ((3 ' - (3- (5-hydroxyhexandihydrocyclopenta [ c ] pyrrol-2 (1H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile
5- (chloromethyl) pyridine-3-carbonitrile hydrochloride (0.33g, 1.65mmol) was dissolved in DMF (12 mL), and cesium carbonate (1.03g, 3.18mmol) was added and stirred at room temperature for 10min. 5-chloro-2-hydroxy-4- ((3 ' - (3- (5-hydroxyhexadihydrocyclopenta [ c ] pyrrol-2 (1H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzaldehyde (0.71g, 1.24mmol) and NaI (19mg, 0.13mmol) were added, the mixture was warmed to 75 ℃ under nitrogen and stirred for 4H. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (100 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and the filtrate concentrated. Silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 460mg of yellow solid in 54.26% yield.
LC-MS：(pos.ion)m/z：666.2[M+1] + ；
Step 3) (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (5-hydroxyhexahydro-dihydrocyclopenta [ c ] pyrrole-2 (1H) -propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
((4-chloro-2-formyl-5- ((3 ' - (3- (5-hydroxyhexandihydrocyclopenta [ c ] pyrrol-2 (1H) -yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile (460mg, 0.69mmol) and D-pipecolic acid (0.033g, 1.03mmol) were dissolved in DMF (15.0 mL), acetic acid was added to adjust the pH to about 5, heated to 60 ℃ and stirred for 1H, cooled to room temperature, sodium cyanoborohydride (0.216g, 3.44mmol) was slowly added, blanketed with nitrogen, and stirred for 12H at room temperature. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated. Purification by silica gel column chromatography (DCM/MeOH =8/1,v/v) gave 80mg of a yellow solid with 14.87% yield.
LC-MS：(pos.ion)m/z：779.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(s，2H)，8.46(s，1H)，7.49(d，J＝7.3Hz，1H)，7.42(s，1H)，7.24(dt，J＝15.8，7.7Hz，2H)，7.13(s，1H)，7.06(d，J＝7.5Hz，1H)，6.95(d，J＝8.1Hz，1H)，6.69(d，J＝7.4Hz，1H)，5.30(d，J＝28.4Hz，4H)，4.07(d，J＝6.3Hz，3H)，3.79(d，J＝13.6Hz，2H)，3.63(d，J＝13.6Hz，2H)，3.51(s，1H)，3.16-3.12(m，1H)，3.02(s，4H)，2.89(s，1H)，2.73(s，2H)，2.30(s，1H)，2.09(s，2H)，2.03(s，3H)，1.83(s，6H)，1.73(s，1H)，1.58-1.44(m，5H)，1.37(s，1H).
Example 11 (S) -1- (4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzyl) piperidine-2-carboxylic acid (Compound 11)
Step 1) 4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
Reacting 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl]-2-methyl-phenyl]Methoxy radical]-5-chloro-2-hydroxy-benzaldehyde (2.8g, 5.6mmol) and 3-oxa-8-azaspiro [4.5]Decane (940mg, 6.65mmol) was dissolved in DMF (12 mL), and K was added successively 2 CO 3 (1.9g, 14mmol) and NaI (1g, 6.67mmol) were heated to 75 ℃ and reacted for 12.5h. The heating was stopped, cooled to room temperature, diluted with water (80 mL), extracted with ethyl acetate (50 mL. Times.3), the organic phase collected, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (DCM/MeOH =8/1,v/v) gave 600mg of a tan viscous mass, 19% yield.
LC-MS：(pos.ion)m/z：564.1[M+1] + ；
Step 2) 4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzaldehyde
Mixing 4- ((3' - (3- (2-oxa-8-azaspiro [ 4.5))]Decan-8-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (550 m)g,0.97 mmol) and 3- (bromomethyl) pyridine hydrobromide (320mg, 1.26mmol) were dissolved in DMF (15 mL) and Cs was added 2 CO 3 (794 mg, 2.43mmol), and NaI (14mg, 0.10mmol), heated to 75 ℃ for reaction for 3.5h. The reaction was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (30 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 380mg of yellow solid in 59.48% yield.
LC-MS：(pos.ion)m/z：655.1[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzyl) piperidine-2-carboxylic acid
4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzaldehyde (120mg, 0.18mmol) and D-pipecolic acid (35mg, 0.27mmol) were dissolved in DMF (8 mL), acetic acid was added to adjust the solution to pH 5 or so, the solution was heated to 60 ℃ and stirred for 1h, cooled to room temperature, sodium cyanoborohydride (57mg, 0.90mmol) was slowly added and the reaction stirred for 15h at room temperature. Stirring was stopped, cooled to room temperature, and saturated carbonic acid solution (30 mL) was added thereto and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (DCM/MeOH =8/1,v/v) to isolate 30mg of a white solid with a yield of 21.32%.
LC-MS：(pos.ion)m/z：768.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ8.70(s，1H)，8.56(d，J＝4.3Hz，1H)，7.90(d，J＝7.7Hz，1H)，7.49(d，J＝7.5Hz，1H)，7.43(s，2H)，7.27(t，J＝7.4Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.14(s，1H)，7.07(d，J＝7.4Hz，1H)，6.96(d，J＝8.1Hz，1H)，6.69(d，J＝7.4Hz，1H)，5.26(d，J＝6.6Hz，4H)，4.06(d，J＝6.3Hz，2H)，3.75-3.69(m，3H)，3.62(d，J＝13.9Hz，2H)，3.51(s，1H)，3.44(s，2H)，3.15(s，1H)，2.89(s，2H)，2.73(s，4H)，2.29(s，1H)，2.03(s，5H)，1.83(s，3H)，1.80-1.76(m，1H)，1.69(s，3H)，1.64(s，3H)，1.47(s，3H)，1.36(s，1H).
Example 12- (3- ((3 ' - ((2-chloro-4- (morpholinomethyl) -5- (pyridin-3-ylmethoxy) phenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2-oxa-8-aza-spiro [4.5] decane (Compound 12)
Step 1) 4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
Reacting 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl]-2-methyl-phenyl]Methoxy radical]-5-chloro-2-hydroxy-benzaldehyde (2.8g, 5.6mmol) and 3-oxa-8-azaspiro [4.5]Decane (940mg, 6.65mmol) was dissolved in DMF (12 mL), and K was added successively 2 CO 3 (1.9g, 14mmol) and NaI (1g, 6.67mmol). Then heated to 75 ℃ and reacted for 12.5h. The heating was stopped, cooled to room temperature, diluted with water (80 mL), extracted with ethyl acetate (50 mL. Times.3), the organic phase collected, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (DCM/MeOH =8/1,v/v) gave 600mg tan thick paste in 19% yield.
LC-MS：(pos.ion)m/z：564.1[M+1] + ；
Step 2) 4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzaldehyde
Mixing 4- ((3' - (3- (2-oxa-8-azaspiro [ 4.5))]Decan-8-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (550mg, 0.97mmol) and 3- (bromomethyl) pyridine hydrobromide (320mg, 1.26mmol) were dissolved in DMF (15 mL) and Cs was added 2 CO 3 (794 mg, 2.43mmol), and NaI (14mg, 0.10mmol), heated to 75 ℃ for reaction for 3.5h. The reaction was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (30 mL. Times.3), and the organic phases combined, free ofDried over sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 380mg of yellow solid in 59.48% yield.
LC-MS：(pos.ion)m/z：655.1[M+1] + ；
Step 3) 8- (3- ((3 ' - ((2-chloro-4- (morpholinomethyl) -5- (pyridin-3-ylmethoxy) phenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2-oxa-8-aza-spiro [4.5] decane
4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzaldehyde (130mg, 0.20mmol) and morpholine (24mg, 0.27mmol) were dissolved in DMF (8 mL), acetic acid was added to adjust the solution pH to around 5, the reaction was stirred at 60 ℃ for 1h, cooled to room temperature, sodium cyanoborohydride (62mg, 0.98mmol) was slowly added, and the reaction was stirred at room temperature for 15h. Stirring was stopped, cooled to room temperature, added with saturated carbonic acid solution (30 mL), stirred at room temperature for 30min, extracted with ethyl acetate (100 mL × 3), combined organic phases were dried over anhydrous sodium sulfate, and separated and purified by silica gel column chromatography (DCM/MeOH =10/1,v/v) to isolate 40mg of white solid with a yield of 27.76%.
LC-MS：(pos.ion)m/z：726.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ8.72(s，1H)，8.55(d，J＝3.4Hz，1H)，7.90(d，J＝7.4Hz，1H)，7.49(d，J＝7.4Hz，1H)，7.47-7.40(m，1H)，7.31(s，1H)，7.26(d，J＝7.4Hz，1H)，7.21(t，J＝7.7Hz，1H)，7.14(s，1H)，7.07(d，J＝7.3Hz，1H)，6.96(d，J＝8.1Hz，1H)，6.69(d，J＝7.3Hz，1H)，5.25(s，4H)，4.06(d，J＝5.7Hz，2H)，3.72(t，J＝6.5Hz，2H)，3.55(s，3H)，3.42(d，J＝13.5Hz，4H)，3.34(s，6H)，2.73(s，3H)，2.33(s，4H)，2.03(s，4H)，1.83(s，3H)，1.67(d，J＝22.4Hz，5H).
Example 13 (R) -2- ((4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzyl) amino) propanoic acid (Compound 13)
Step 1) 4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
Reacting 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl]-2-methyl-phenyl]Methoxy radical]-5-chloro-2-hydroxy-benzaldehyde (2.8g, 5.6mmol) and 3-oxa-8-azaspiro [4.5]Decane (940mg, 6.65mmol) was dissolved in DMF (12 mL), and K was added successively 2 CO 3 (1.9g, 14mmol) and NaI (1g, 6.67mmol). Then heated to 75 ℃ and reacted for 12.5h. The heating was stopped, cooled to room temperature, diluted with water (80 mL), extracted with ethyl acetate (50 mL. Times.3), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (DCM/MeOH =8/1,v/v) gave 600mg of a tan viscous mass, 19% yield.
LC-MS：(pos.ion)m/z：564.1[M+1] + ；
Step 2) 4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzaldehyde
Mixing 4- ((3' - (3- (2-oxa-8-azaspiro [ 4.5))]Decan-8-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (550mg, 0.97mmol) and 3- (bromomethyl) pyridine hydrobromide (320mg, 1.26mmol) were dissolved in DMF (15 mL) and Cs was added 2 CO 3 (794 mg, 2.43mmol) and NaI (14mg, 0.097mmol) and subsequently heated to 75 ℃ for 3.5h. The reaction was stopped, cooled to room temperature, diluted with water (30 mL), extracted with ethyl acetate (30 mL. Times.3), the organic phases combined, dried over anhydrous sodium sulfate, filtered and concentrated. Purification by silica gel column chromatography (DCM/MeOH =10/1,v/v) gave 380mg of yellow solid in 59.48% yield.
LC-MS：(pos.ion)m/z：655.1[M+1] + ；
Step 3) (R) -2- ((4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzyl) amino) propanoic acid
4- ((3 ' - (3- (2-oxa-8-azaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzaldehyde (120mg, 0.183mmol) and (2R) -2-aminopropionic acid (24mg, 0.26mmol) were dissolved in DMF (8 mL), acetic acid was added to adjust the solution pH to around 5, heated to 60 ℃ and stirred for 1h, cooled to room temperature, sodium cyanoborohydride (57mg, 0.90mmol) was slowly added and stirred for 15h at room temperature. Stirring was stopped, cooled to room temperature, saturated carbonic acid solution (30 mL) was added thereto and stirred at room temperature for 30min, extracted with ethyl acetate (100 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, and subjected to silica gel column chromatography (DCM/MeOH =8/1,v/v) to isolate 15mg of a white solid with a yield of 11.25%.
LC-MS：(pos.ion)m/z：728.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ8.74(s，1H)，8.57(d，J＝4.1Hz，1H)，7.97(s，1H)，7.54(s，1H)，7.49(d，J＝7.0Hz，1H)，7.44(dd，J＝7.8，4.9Hz，1H)，7.27(t，J＝7.5Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.20-7.13(m，1H)，7.07(d，J＝7.5Hz，1H)，6.96(d，J＝8.4Hz，1H)，6.69(d，J＝7.6Hz，1H)，5.39-5.19(m，4H)，4.07(d，J＝6.9Hz，2H)，3.94-3.85(m，2H)，3.73(t，J＝7.0Hz，2H)，3.51(s，2H)，3.45(s，3H)，3.19-3.14(m，2H)，2.89(s，6H)，2.10(s，2H)，2.03(s，3H)，1.82(d，J＝8.0Hz，3H)，1.70(d，J＝7.0Hz，6H).
Example 14 (S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 14)
Step 1) 5-chloro-2-hydroxy-4- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (700mg, 1.39mmol) and 2-hydroxy-2,7-diazaspiro [3.5] nonane hydrochloride (370.3mg, 2.08mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (576.1mg, 4.17mmol) and sodium iodide (312.4mg, 2.08mmol) in that order, followed by heating to 70 ℃ for 16h. Stopping stirring, cooling to room temperature, and completely reacting the raw materials. Water dilution (20 mL), ethyl acetate extraction (50 mL × 3), organic phases combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 685mg of a yellow solid in 87.4% yield.
LC-MS：(pos.ion)m/z：564.0[M+1] + ；
Step 2) 5- ((4-chloro-2-formyl-5- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile
To 5-chloro-2-hydroxy-4- ((3' - (3- (2-hydroxy-7-azaspiro [3.5]]Nonan-7-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) methoxy) benzaldehyde (680mg, 1.21mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (273.5mg, 1.45mmol) in DMF (20 mL), cesium carbonate (981.9mg, 3.01mmol) and sodium iodide (36.14mg, 0.24mmol), N, were added in this order 2 Protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 476mg of a yellow solid in 58.05% yield.
LC-MS：(pos.ion)m/z：680.2[M+1] + ；
Step 3) (S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((4-chloro-2-formyl-5- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile (250mg, 0.37mmol) and D-pipecolic acid (94.94mg, 0.74mmol) in DMF (10 mL) was added acetic acid, the pH was adjusted to about 5, reacted at 60 ℃ for 1h, cooled to room temperature, then added sodium borocyanide (115.5mg, 1.84mmol) to the reaction system, reacted at room temperature for 3h, and reacted at 80 ℃ for 16h. Stirring was stopped and then saturated potassium carbonate solution was added and stirred for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 70mg of a pale yellow solid with a yield of 24.01%.
LC-MS：(pos.ion)m/z：793.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(s，1H)，8.99(s，1H)，8.45(s，1H)，7.49(d，J＝7.4Hz，1H)，7.41(s，1H)，7.26(t，J＝7.5Hz，1H)，7.20(t，J＝7.9Hz，1H)，7.11(s，1H)，7.06(d，J＝7.4Hz，1H)，6.94(d，J＝8.1Hz，1H)，6.67(d，J＝7.4Hz，1H)，5.32(s，2H)，5.26(s，2H)，4.05(dd，J＝16.0，7.0Hz，4H)，3.76(d，J＝13.9Hz，2H)，3.60(d，J＝13.9Hz，3H)，3.51(s，2H)，3.14(d，J＝6.9Hz，2H)，2.11-2.04(m，2H)，2.02(s，3H)，1.92(s，2H)，1.81(s，4H)，1.51(dd，J＝14.6，7.9Hz，9H)，1.36(s，1H)，1.23(s，3H).
Example 15 (S) -1- (4- ((3 ' - (3- (2-oxa-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 15)
Step 1) 4- ((3 ' - (3- (2-oxa-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (500mg, 0.99mmol) and 2-oxo-7-azaspiro [3.5] nonane oxalate (323.4 mg, 1.49mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (617.3 mg, 4.47mmol) and NaI (223.1mg, 1.49mmol) in that order, and the mixture was heated to 70 ℃ for 16h. Stopping stirring, cooling to room temperature, and completely reacting the raw materials. Water dilution (20 mL), ethyl acetate extraction (50 mL × 3), organic phases combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 307mg of yellow solid in 56.23% yield.
LC-MS：(pos.ion)m/z：550.1[M+1] + ；
Step 2) 5- ((5- ((3 ' - (3- (2-oxa-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -pyridin-3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (2-oxa-7-azaspiro [3.5 ])]Nonan-7-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]To a solution of (3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (300mg, 0.55mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (1546 mg, 0.82mmol) in DMF (20 mL) were added cesium carbonate (309.9mg, 1.2mmol) and sodium iodide (16.35mg, 0.11mmol), N, in that order 2 Protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH = 10/1) to give 73mg of yellow oily liquid in 20.09% yield.
LC-MS：(pos.ion)m/z：666.4[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (2-oxa-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (2-oxa-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -pyridin-3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (70mg, 0.111mmol) and D-pipecolic acid (27.14mg, 0.21mmol) in DMF (10 mL) was added acetic acid, the pH was adjusted to around 5, reacted at 60 ℃ for 1h, cooled to room temperature, then sodium borocyanide (33.01mg, 0.53mmol) was added to the reaction system, reacted at room temperature for 3h, and then warmed to 80 ℃ for 16h. Then, the stirring was stopped, and a saturated potassium carbonate solution was added thereto and stirred for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 15mg of a light yellow solid in 18.32% yield.
LC-MS：(pos.ion)m/z：779.5[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(d，J＝7.7Hz，2H)，8.46(s，1H)，7.49(d，J＝7.5Hz，1H)，7.42(s，1H)，7.32-7.16(m，2H)，7.12(s，1H)，7.07(d，J＝7.4Hz，1H)，6.96(d，J＝8.2Hz，1H)，6.69(d，J＝7.5Hz，1H)，5.32(d，J＝14.0Hz，2H)，5.27(s，2H)，4.29(s，3H)，4.05(d，J＝6.4Hz，2H)，3.77(d，J＝13.5Hz，2H)，3.61(d，J＝13.6Hz，1H)，3.52(s，6H)，2.03(s，3H)，1.82(s，4H)，1.49(s，3H)，1.24(s，9H)，0.85(d，J＝7.0Hz，2H).
Example 16 (S) -1- (4- ((3 ' - (3- (2-oxa-6-azaspiro [3.5] non-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 16)
Step 1) 4- ((3 ' - (3- (2-oxa-6-azaspiro [3.5] non-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (600mg, 1.19mmol) and 2-oxo-6-azaspiro [3.5] nonane oxalate (388mg, 1.79mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (740.7mg, 5.36mmol) and NaI (267.8mg, 1.79mmol) in that order, and the mixture was heated to 70 ℃ for 16h. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 427mg of a red oily liquid in 65.18% yield.
LC-MS：(pos.ion)m/z：550.1[M+1] + ；
Step 2) 5- ((5- ((3 ' - (3- (2-oxa-6-azaspiro [3.5] non-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -pyridin-3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (2-oxa-6-azaspiro [3.5 ])]Nonyl-6-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]To a solution of (420mg, 0.76mmol) of-3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde and (173.2mg, 0.92mmol) of 5-chloromethyl-3-cyanopyridine hydrochloride in DMF (20 mL) were added cesium carbonate (621.9mg, 1.91mmol) and sodium iodide (22.89mg, 0.15mmol), N 2 Protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 342mg of red oily liquid in 67.24% yield.
LC-MS：(pos.ion)m/z：666.5[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (2-oxa-6-azaspiro [3.5] non-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (2-oxa-6-azaspiro [3.5] non-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -pyridin-3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (340mg, 0.51mmol) and D-pipecolic acid (131.8mg, 1.02mmol) in DMF (20 mL) was added acetic acid to adjust the pH to around 5, followed by heating to 60 ℃ for reaction for 1 hour, cooling to room temperature, adding sodium borocyanide (160.4mg, 2.55mmol) to the reaction system, stirring at room temperature for reaction for 3 hours, and then heating to 80 ℃ for reaction for 16 hours. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (50 mL × 3), organic phases combined, organic phase washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, column chromatographically isolated purification (DCM/MeOH =6/1,v/v) to yield light red solid 52mg, 13.07% yield.
LC-MS：(pos.ion)m/z：779.6[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(s，2H)，8.48(s，1H)，7.46(d，J＝18.6Hz，2H)，7.23(d，J＝23.1Hz，2H)，7.15(s，1H)，7.07(s，1H)，6.97(s，1H)，6.69(s，1H)，5.31(d，J＝31.0Hz，4H)，4.26(d，J＝31.9Hz，5H)，4.05(s，4H)，3.81(s，6H)，2.00(d，J＝18.4Hz，6H)，1.83(s，4H)，1.75(s，3H)，1.50(s，5H)，1.37(s，1H)，1.17(s，1H).
Example 17 (2S) -1- (4- ((3 ' - (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 17)
Step 1) 4- ((3 ' - (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (600mg, 1.191mmol) and 2-oxo-7-azaspiro [4.5] decane hydrochloride (317.4mg, 1.79mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (411.5mg, 2.98mmol) and NaI (267.8mg, 1.79mmol) in this order, and the mixture was heated to 70 ℃ for 16 hours. The stirring was stopped and the mixture was cooled to room temperature. Then, water was added to dilute (20 mL), ethyl acetate was extracted (50 mL × 3), the organic phases were combined, and the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to obtain 200mg of a brownish red oily liquid with a yield of 29.77%.
LC-MS：(pos.ion)m/z：564.4[M+1] + ；
Step 2) 5- ((5- ((3 ' - (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -pyridin-3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (2-oxa-7-azaspiro [4.5 ])]Decan-7-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]To a DMF solution (20 mL) of (4-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (450mg, 0.80mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (181mg, 0.96mmol) were added cesium carbonate (649.8mg, 1.99mmol) and sodium iodide (23.92mg, 0.16mmol), N 2 Protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 520mg of red oily liquid with a yield of 95.83%.
LC-MS：(pos.ion)m/z：680.5[M+1] + ；
Step 3) (2S) -1- (4- ((3 ' - (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -pyridin-3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (520mg, 0.76mmol) and D-pipecolic acid (197.5 mg, 1.53mmol) in DMF (20 mL) was added acetic acid, the pH was adjusted to about 5, reacted at 60 ℃ for 1 hour, cooled to room temperature, then added sodium borocyanide (240.2mg, 3.82mmol) to the reaction system, reacted at room temperature for 3 hours, and reacted at 80 ℃ for 16 hours. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 112mg of a pale red solid in 18.47% yield.
LC-MS：(pos.ion)m/z：793.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(d，J＝1.3Hz，2H)，8.47(s，1H)，7.49(d，J＝7.5Hz，1H)，7.43(s，1H)，7.26(t，J＝7.6Hz，1H)，7.20(t，J＝7.9Hz，1H)，7.15(s，1H)，7.06(d，J＝7.5Hz，1H)，6.96(d，J＝8.2Hz，1H)，6.68(d，J＝7.5Hz，1H)，5.35(s，2H)，5.27(s，2H)，4.06(d，J＝6.2Hz，3H)，3.84(s，3H)，3.66(d，J＝13.9Hz，9H)，2.03(s，4H)，1.82(s，5H)，1.71(d，J＝9.7Hz，2H)，1.50(t，J＝48.5Hz，9H)，1.23(s，2H).
Example 18 (S) -1- (4- ((3 ' - (3- (9-oxa-2-azaspiro [5.5] undecan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 18)
Step 1) 4- ((3 ' - (3- (9-oxa-2-azaspiro [5.5] undecan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (800mg, 1.59mmol) and 9-oxo-2-azaspiro [5.5] undecane (369.8mg, 2.38mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (329.2mg, 2.38mmol) and NaI (357mg, 2.38mmol) in that order, and the mixture was heated to 70 ℃ for 16h. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 566mg of a brownish red oily liquid with a yield of 61.66%.
LC-MS：(pos.ion)m/z：578.4[M+1] + ；
Step 2) 5- ((5- ((3 ' - (3- (9-oxa-2-azaspiro [5.5] undecan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (9-oxa-2-azaspiro [5.5 ]))]Undecan-2-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]To a solution of (560mg, 0.97mmol) of-3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde and (219.7mg, 1.16mmol) of 5-chloromethyl-3-cyanopyridine hydrochloride in DMF (20 mL) were added cesium carbonate (789 mg, 2.42mmol) and sodium iodide (29.04mg, 0.19mmol), N 2 And (4) protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =20/1,v/v) to give 408mg of red oily liquid with a yield of 60.66%.
LC-MS：(pos.ion)m/z：694.5[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (9-oxa-2-azaspiro [5.5] undecan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (9-oxa-2-azaspiro [5.5] undecan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (300mg, 0.43mmol) and D-pipecolic acid (111.6 mg, 0.86mmol) in DMF (20 mL) was added acetic acid, the pH was adjusted to about 5, the reaction was carried out at 60 ℃ for 1 hour, cooled to room temperature, sodium borocyanide (135.8mg, 2.16mmol) was added to the reaction system, and after 3 hours at room temperature, the reaction was carried out at 80 ℃ for 16 hours. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 18mg of a pale red solid in 5.16% yield.
LC-MS：(pos.ion)m/z：807.6[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.00(d，J＝6.1Hz，2H)，8.46(s，1H)，7.49(d，J＝7.3Hz，1H)，7.42(s，1H)，7.25(dt，J＝15.9，7.7Hz，2H)，7.12(d，J＝2.7Hz，1H)，7.06(d，J＝7.4Hz，1H)，6.97(d，J＝8.3Hz，1H)，6.70(d，J＝7.5Hz，1H)，5.33(s，2H)，5.27(s，2H)，4.13-4.03(m，2H)，3.81(d，J＝13.6Hz，2H)，3.65(d，J＝14.0Hz，3H)，3.52(d，J＝12.6Hz，6H)，2.06-1.95(m，5H)，1.84(d，J＝3.7Hz，4H)，1.71(s，4H)，1.48(d，J＝9.4Hz，4H)，1.39(s，2H)，1.23(s，6H)，0.85(s，1H).
Example 19 (S) -1- (4- ((3 ' - (3- (8-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 19)
Step 1) 4- ((3 ' - (3- (8-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (800mg, 1.59mmol) and 8-oxa-2-azaspiro [4.5] decane (336.3mg, 2.38mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (329.2mg, 2.38mmol) and NaI (357mg, 2.38mmol) in that order, and the mixture was heated to 70 ℃ for 16 hours. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 640mg of a brownish red oily liquid with a yield of 71.45%.
LC-MS：(pos.ion)m/z：564.2[M+1] +
Step 2) 5- ((5- ((3 ' - (3- (8-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (8-oxa-2-azaspiro [4.5 ])]Dec-2-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]To a solution of (3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (640mg, 1.14mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (257.4mg, 1.36mmol) in DMF (20 mL) were added cesium carbonate (924.1mg, 2.83mmol) and sodium iodide (34.01mg, 0.23mmol), N, in that order 2 Protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 190mg of red oily liquid in 24.62% yield.
LC-MS：(pos.ion)m/z：680.1[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (8-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (8-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (190mg, 0.28mmol) and D-pipecolic acid (72.16mg, 0.56mmol) in DMF (10 mL) was added acetic acid, the pH was adjusted to around 5, followed by heating to 60 ℃ for reaction for 1h, cooling to room temperature, adding sodium borocyanide (87.77mg, 1.40mmol) to the reaction system, stirring at room temperature for 3h, and heating to 80 ℃ for reaction for 16h. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 35mg of a light yellow solid in 15.79% yield.
LC-MS：(pos.ion)m/z：793.3[M+1] + ；
1 H NMR(40)0MHz，d 6 -DMSO)δ9.04-8.96(m，2H)，8.46(s，1H)，7.49(d，J＝7.4Hz，1H)，7.42(s，1H)，7.26(t，J＝7.5Hz，1H)，7.18(d，J＝7.8Hz，1H)，7.12(s，1H)，7.06(d，J＝7.5Hz，1H)，6.95(d，J＝8.2Hz，1H)，6.68(d，J＝7.5Hz，1H)，5.32(d，J＝11.3Hz，2H)，5.25(d，J＝12.6Hz，2H)，3.83-3.75(m，3H)，3.64(s，1H)，3.60(s，1H)，3.50(t，J＝10.8Hz，6H)，3.15(dd，J＝11.3，4.3Hz，1H)，2.89(s，3H)，2.62(s，2H)，2.02(s，3H)，1.82(s，5H)，1.68(t，J＝6.7Hz，3H)，1.49(t，J＝13.0Hz，7H)，1.36(s，1H)，1.22(s，1H).
Example 20 (2S) -1- (4- ((3 ' - (3- (7-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 20)
Step 1) 4- ((3 ' - (3- (7-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxybenzaldehyde (800mg, 1.59mmol) and 7-oxa-2-azaspiro [4.5] decane hemioxalate (414.1mg, 1.11mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (548.7 mg, 3.97mmol) and NaI (357 mg, 2.38mmol) in this order, heated to 70 ℃ and reacted for 16h. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), and the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH = 10/1) to give 548mg of a brownish red oily liquid, yield 61.18%.
LC-MS：(pos.ion)m/z：564.4[M+1] +
Step 2) 5- ((5- ((3 ' - (3- (7-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (7-oxa-2-azaspiro [4.5 ])]Dec-2-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]To a solution of (3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (540mg, 0.96mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (221.2mg, 1.17mmol) in DMF (20 mL) were added cesium carbonate (794.2mg, 2.44mmol) and sodium iodide (29.23mg, 0.19mmol), N 2 Protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =20/1,v/v) to give 616mg of yellow oily liquid with 94.6% yield.
LC-MS：(pos.ion)m/z：680.2[M+1] + ；
Step 3) (2S) -1- (4- ((3 ' - (3- (7-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (7-oxa-2-azaspiro [4.5] decan-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (300mg, 0.44mmol) and D-pipecolic acid (113.9mg, 0.88mmol) in DMF (10 mL) was added acetic acid, the pH was adjusted to about 5, the reaction was carried out at 60 ℃ for 1 hour, the reaction was cooled to room temperature, sodium borocyanide (138.6mg, 2.12mmol) was added to the reaction system, and after 3 hours at room temperature, the reaction was carried out at 80 ℃ for 16 hours. Stirring was stopped, saturated potassium carbonate solution was added and stirred for 30min. Then, water was added to dilute (20 mL), ethyl acetate was extracted (20 mL × 3), the organic phases were combined, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to obtain 48mg of a pale red solid with a yield of 13.72%.
LC-MS：(pos.ion)m/z：793.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.03-8.97(m，2H)，8.46(s，1H)，7.49(d，J＝7.4Hz，1H)，7.42(s，1H)，7.27(t，J＝7.5Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.12(s，1H)，7.07(d，J＝7.3Hz，1H)，6.96(d，J＝8.1Hz，1H)，6.69(d，J＝7.5Hz，1H)，5.33(s，2H)，5.27(s，2H)，4.07(d，J＝6.5Hz，3H)，3.78(d，J＝13.9Hz，2H)，3.64(s，4H)，3.51(s，4H)，2.03(s，5H)，1.83(s，4H)，1.64(s，2H)，1.52(d，J＝25.4Hz，9H)，1.38(d，J＝7.0Hz，1H)，1.24(s，3H).
Example 160 (2S) -1- (4- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 160)
Step 1) 4- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (600mg, 1.91mmol) and 2-oxa-7-azaspiro [4.4] nonane (227.2mg, 1.79mmol) in DMF (20 mL) under nitrogen was added potassium carbonate (246.9mg, 1.79mmol) and NaI (267.8mg, 1.79mmol) in that order, and the mixture was heated to 70 ℃ for 16h. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 200mg of a brownish red oily liquid with a yield of 30.53%.
LC-MS：(pos.ion)m/z：550.4[M+1] +
Step 2) 5- ((5- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (2-oxa-7-azaspiro [4.4 ])]Nonan-7-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) methoxy) -5-chloro-2-hydroxybenzaldehydeTo a solution of 5-chloromethyl-3-cyanopyridine hydrochloride (226.8mg, 1.2mmol) in DMF (20 mL) (550mg, 0.99mmol) and cesium carbonate (814.4mg, 2.5mmol) and sodium iodide (29.97mg, 0.2mmol), N, in that order, were added 2 Protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 274mg of a yellow solid in 41.14% yield.
LC-MS：(pos.ion)m/z：666.2[M+1] + ；
Step 3) (2S) -1- (4- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] nonan-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (270mg, 0.40mmol) and D-pipecolic acid (104.7 mg, 0.81mmol) in DMF (10 mL) was added acetic acid, the pH was adjusted to about 5, followed by heating to 60 ℃ for 1 hour, cooling to room temperature, adding sodium borocyanide (127.3mg, 2.02mmol) to the reaction system, reacting at room temperature for 3 hours, and reacting at 80 ℃ for 16 hours. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 53mg of a light yellow solid in 16.78% yield.
LC-MS：(pos.ion)m/z：779.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.00(d，J＝2.8Hz，2H)，8.45(s，1H)，7.49(d，J＝7.4Hz，1H)，7.42(s，1H)，7.26(t，J＝7.6Hz，1H)，7.20(t，J＝7.9Hz，1H)，7.11(s，1H)，7.06(d，J＝7.4Hz，1H)，6.95(d，J＝8.2Hz，1H)，6.68(d，J＝7.4Hz，1H)，5.33(s，2H)，5.26(s，2H)，4.05(dt，J＝12.9，6.5Hz，3H)，3.79(d，J＝13.7Hz，2H)，3.73-3.68(m，3H)，3.62(d，J＝13.7Hz，1H)，3.55(s，1H)，3.51(s，1H)，3.44(d，J＝8.1Hz，1H)，3.17(s，1H)，2.03(s，3H)，1.99-1.93(m，3H)，1.89(d，J＝7.1Hz，1H)，1.87-1.75(m，9H)，1.73(s，1H)，1.48(s，3H)，1.36(s，1H)，1.23(s，1H).
Example 22 (S) -1- (4- ((3 ' - (3- (2-oxa-6-azaspiro [3.4] oct-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 22)
Step 1) 4- ((3 ' - (3- (2-oxa-6-azaspiro [3.4] oct-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (600mg, 1.19mmol) and 2-oxa-6-azaspiro [3.4] octane (263.8mg, 0.83mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (411.5mg, 2.97mmol) and NaI (267.8mg, 1.78mmol) in that order, and the mixture was heated to 70 ℃ for 16 hours. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 260mg of red oily liquid with a yield of 40.72%.
LC-MS：(pos.ion)m/z：536.2[M+1] + ；
Step 2) 5- ((5- ((3 ' - (3- (2-oxa-6-azaspiro [3.4] oct-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (2-oxa-6-azaspiro [3.4 ])]Oct-6-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (260mg, 0.48mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (110mg,0.58 mmol) in DMF (20 mL) was added cesium carbonate (395mg, 1.21mmol) and sodium iodide (14.54mg, 0.10mmol), in that order, and N 2 Protecting, heating to 75 ℃ and reacting for 4h. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 240mg of red oily liquid in 75.88% yield.
LC-MS：(pos.ion)m/z：652.5[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (2-oxa-6-azaspiro [3.4] oct-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (2-oxa-6-azaspiro [3.4] oct-6-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (240mg, 0.37mmol) and D-pipecolic acid (95.06mg, 0.74mmol) in DMF (10 mL) was added acetic acid, the pH was adjusted to about 5, followed by heating to 60 ℃ for reaction for 1 hour, cooling to room temperature, adding sodium borocyanide (115.mg, 1.84mmol) to the reaction system, stirring at room temperature for 3 hours, and then heating to 80 ℃ for reaction for 16 hours. Stirring was stopped, saturated potassium carbonate solution was added and stirred for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 31mg of a white solid in 11.01% yield.
LC-MS：(pos.ion)m/z：765.5[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(s，2H)，8.48(s，1H)，7.49(d，J＝7.6Hz，1H)，7.44(s，1H)，7.30-7.14(m，3H)，7.06(d，J＝7.3Hz，1H)，6.95(d，J＝8.1Hz，1H)，6.67(d，J＝7.4Hz，1H)，5.36(s，2H)，5.28(s，2H)，4.49(dd，J＝19.7，5.6Hz，4H)，4.04(dd，J＝14.1，6.8Hz，3H)，3.83(d，J＝13.7Hz，2H)，3.67(d，J＝13.6Hz，4H)，2.72(s，2H)，2.13(s，2H)，2.00(d，J＝17.2Hz，6H)，1.81(s，4H)，1.72(s，1H)，1.50(s，3H)，1.36(s，1H)，1.22(s，1H).
Example 23 (S) -1- (4- ((3 ' - (3- (7-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 23)
Step 1) 4- ((3 ' - (3- (7-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (1000mg, 1.98mmol) and 7-oxo-2-azaspiro [3.5] nonane hemioxalate (478.5mg, 1.39mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (685.8mg, 4.96mmol) and NaI (446.3mg, 2.98mmol) in this order, and the mixture was heated to 70 ℃ for 16 hours. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 553mg of a brownish red oily liquid, 50.65% yield.
LC-MS：(pos.ion)m/z：550.4[M+1] + ；
Step 2) 5- ((5- ((3 ' - (3- (7-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (7-oxa-2-azaspiro [3.5 ]))]Nonan-2-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]To a solution of (3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (550mg, 0.10mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (226.8mg, 1.2mmol) in DMF (20 mL) were added cesium carbonate (814.4mg, 2.5mmol) and sodium iodide (29.97mg, 0.20mmol), N-in that order 2 Protection, heating toThe reaction is carried out for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 220mg of red oily liquid with a yield of 33.03%.
LC-MS：(pos.ion)m/z：666.2[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (7-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (7-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (220mg, 0.33mmol) and D-pipecolic acid (85.31mg, 0.66mmol) in DMF (20 mL) was added acetic acid, the pH was adjusted to about 5, followed by heating to 60 ℃ for 1h, cooling to room temperature, adding sodium borocyanide (103.8mg, 1.652mmol) to the reaction system, reacting at room temperature for 3h, and reacting at 80 ℃ for 16h. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 20mg of a light yellow solid in 7.77% yield.
LC-MS：(pos.ion)m/z：779.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(s，2H)，8.47(s，1H)，7.49(d，J＝7.2Hz，1H)，7.42(s，1H)，7.26(t，J＝7.5Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.14(s，1H)，7.06(d，J＝7.5Hz，1H)，6.96(d，J＝8.2Hz，1H)，6.69(d，J＝7.5Hz，1H)，5.35(s，2H)，5.27(s，2H)，4.07(d，J＝6.0Hz，3H)，3.81(s，1H)，3.77(d，J＝5.0Hz，1H)，3.68-3.58(m，5H)，2.02(d，J＝6.3Hz，3H)，1.99-1.88(m，3H)，1.82(s，4H)，1.75(s，6H)，1.49(s，4H)，1.36(s，1H)，1.23(s，4H).
Example 24 (S) -1- (4- ((3 ' - (3- (6-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 24)
Step 1) 4- ((3 ' - (3- (6-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (1000mg, 1.98mmol) and 6-oxo-2-azaspiro [3.5] nonane hemioxalate (478.5mg, 1.39mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (685.8mg, 4.96mmol) and NaI (446.3mg, 2.97mmol) in this order, and the mixture was heated to 70 ℃ for 16 hours. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 356mg of red oily liquid in 32.60% yield.
LC-MS：(pos.ion)m/z：550.4[M+1] + ；
Step 2) 5- ((5- ((3 ' - (3- (6-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
To 4- ((3' - (3- (6-oxa-2-azaspiro [3.5 ]))]Nonan-2-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]To a solution of (350mg, 0.64mmol) of (3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde and (144.3mg, 0.76mmol) of 5-chloromethyl-3-cyanopyridine hydrochloride in DMF (20 mL) were added cesium carbonate (518.3mg, 1.59mmol) and sodium iodide (19.07mg, 0.13mmol), N, in that order 2 Protecting, and reacting for 4h at 75 ℃. Stirring was stopped, the mixture was cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL. Times.3), the organic phases were combined, washed with saturated brine (20 mL), and dried over anhydrous sodium sulfateDrying, filtration, concentration, column chromatography purification (DCM/MeOH =10/1,v/v) gave 168mg of red oily liquid in 39.63% yield.
LC-MS：(pos.ion)m/z：666.2[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (6-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((5- ((3 ' - (3- (6-oxa-2-azaspiro [3.5] non-2-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (160mg, 0.24mmol) and D-pipecolic acid (62.04mg, 0.66mmol) in DMF (20 mL) was added acetic acid, the pH was adjusted to about 5, followed by heating to 60 ℃ for reaction for 1h, cooling to room temperature, adding sodium borocyanide (62.04mg, 0.98mmol) to the reaction system, and after reaction for 3h at room temperature, reacting for 16h at 80 ℃. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =7/1,v/v) to give 33mg of white solid with a yield of 17.63%.
LC-MS：(pos.ion)m/z：779.4[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.00(d，J＝5.9Hz，2H)，8.45(s，1H)，7.48(d，J＝7.3Hz，1H)，7.41(s，1H)，7.26(t，J＝7.5Hz，1H)，7.19(t，J＝7.9Hz，1H)，7.10(s，1H)，7.06(d，J＝7.6Hz，1H)，6.93(d，J＝8.2Hz，1H)，6.67(d，J＝7.5Hz，1H)，5.29(d，J＝24.5Hz，4H)，4.04-3.96(m，3H)，3.77(d，J＝13.8Hz，2H)，3.61(s，3H)，3.03(d，J＝6.9Hz，2H)，2.59(t，J＝6.8Hz，2H)，2.26(s，1H)，2.00(d，J＝13.8Hz，5H)，1.80(s，4H)，1.74(d，J＝6.7Hz，3H)，1.69-1.63(m，2H)，1.48(s，3H)，1.43(d，J＝5.5Hz，3H)，1.23(s，2H).
Example 25 (S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,9-diazaspiro [5.5] undecan-9-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 25)
Step 1) 5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,9-diazaspiro [5.5] undec-9-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (1000mg, 1.09mmol) and 5,9-diazaspiro [5.5] undecane hydrochloride (609.5mg, 2.978mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (435.2mg, 5.95mmol) and NaI (446.3mg, 2.98mmol) in that order, followed by heating to 70 ℃ for 16h. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 422mg of a yellow solid in 54.29% yield.
LC-MS：(pos.ion)m/z：591.4[M+1] +
Step 2) 5- ((4-chloro-5- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,9-diazaspiro [5.5] undec-9-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-formylphenoxy) methyl) nicotinonitrile
To 5-chloro-4- ((2,2 '-dimethyl-3' - (3- (2-oxo-1,9-diazaspiro [ 5.5)]Undec-9-yl) propoxy) - [1,1' -biphenyl]To a solution of (3-yl) methoxy) -2-hydroxybenzaldehyde (630mg, 1.07mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (241.8mg, 1.28mmol) in DMF (20 mL) were added cesium carbonate (868.2mg, 2.66mmol) and sodium iodide (31.95mg, 0.21mmol), N, in that order 2 Protecting, heating to 75 ℃ and reacting for 4h. Stirring was stopped, cooling to room temperature, diluting with water (20 mL), extracting with ethyl acetate (50 mL. Times.3), combining the organic phases, washing the organic phase with saturated brine (20 mL), drying over anhydrous sodium sulfate, filtering, concentrating, and column chromatographyPurification by isolation (DCM/MeOH =10/1,v/v) gave 422mg of red solid in 55.98% yield.
LC-MS：(pos.ion)m/z：707.2[M+1] + ；
Step 3) (S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,9-diazaspiro [5.5] undecan-9-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of 5- ((4-chloro-5- ((2,2 ' -dimethyl-3 ' - (3- (2-oxo-1,9-diazaspiro [5.5] undec-9-yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-formylphenoxy) methyl) nicotinonitrile (200mg, 0.28mmol) and D-pipecolic acid (73.04mg, 0.56mmol) in DMF (10 mL) was added acetic acid, the pH was adjusted to about 5, followed by heating to 60 ℃ for 1h, cooling to room temperature, adding sodium borocyanide (88.84mg, 0.14mmol) to the reaction system, stirring at room temperature for 3h, and heating to 80 ℃ for 16h. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 22mg of white solid in 9.48% yield.
LC-MS：(pos.ion)m/z：820.6[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.00(dd，J＝7.1，1.7Hz，2H)，8.46(s，1H)，7.49(d，J＝7.5Hz，1H)，7.41(s，1H)，7.27(t，J＝7.7Hz，1H)，7.21(t，J＝7.9Hz，1H)，7.11(s，1H)，7.07(d，J＝7.2Hz，1H)，6.95(d，J＝8.2Hz，1H)，6.68(d，J＝7.5Hz，1H)，5.33(s，2H)，5.26(s，2H)，4.05(d，J＝7.2Hz，2H)，3.77(d，J＝13.8Hz，2H)，3.61(d，J＝13.7Hz，3H)，3.51(s，5H)，3.43-3.39(m，3H)，2.09(s，2H)，2.03(s，3H)，1.82(s，4H)，1.63(s，6H)，1.48(s，3H)，1.37(s，1H)，1.23(s，3H).
Example 26 (2S) -1- (4- ((3 '- (3- (2,7-diazaspiro [4.4] nonan-2-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 26)
Step 1) tert-butyl-7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [4.4] nonane-2-carboxylic acid methyl ester
To a solution of 4- ((3 '- (3-bromopropoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (1000mg, 2.04mmol) and 2,7-diazaspiro [4.4] nonane-2-carboxylic acid tert-butyl ester (693.1mg, 3.06mmol) in DMF (20 mL) under nitrogen protection was added potassium carbonate (42mg, 3.07mmol) followed by sodium iodide (460mg, 3.06mmol), and the mixture was heated to 70 ℃ for 16h. Stirring was stopped, cooled to room temperature, then diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 920mg of red oily liquid in 70.94% yield.
LC-MS：(pos.ion)m/z：635.3[M-1] + ；
Step 2) tert-butyl-7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [4.4] nonane-2-carboxylic acid methyl ester
To tert-butyl-7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2' -methyl- [1,1' -biphenyl]-3-yl) oxy) propyl) -2,7-diazaspiro [4.4]]Methyl nonane-2-carboxylate (920mg, 1.44mmol) and 5-chloromethyl-3-cyanopyridine hydrochloride (328.6mg, 1.73mmol) in DMF (20 mL) were added cesium carbonate (1.18g, 3.62mmol) and sodium iodide (43.42mg, 0.28mmol), N 2 Protecting, heating to 75 ℃ and reacting for 4h. The stirring was stopped and the mixture was cooled to room temperature. Diluting with water (20 mL), extracting with ethyl acetate (50 mL. Times.3), combining the organic phases, washing the organic phase with saturated brine (20 mL), drying over anhydrous sodium sulfate, filtering, concentrating, and purifying by column chromatography (DCM/MeOH =)10/1,v/v) gave 840mg of red oily liquid in 77.19% yield.
LC-MS：(pos.ion)m/z：751.3[M+1] + ；
Step 3) (2S) -1- (4- ((3 '- (3- (7- (tert-butoxycarbonyl) -2,7-diazaspiro [4.4] nonan-2-yl) propoxy) -2-methyl- [1,1' biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of tert-butyl-7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [4.4] nonane-2-carboxylic acid methyl ester (840mg, 1.11mmol) and D-pipecolic acid (288.8mg, 2.23mmol) in DMF (20 mL) was added acetic acid, the pH was adjusted to around 5, the reaction was carried out at 60 ℃ for 1h, cooled to room temperature, and sodium borocyanide (351.3mg, 5.59mmol) was added to the reaction system, followed by heating to 80 ℃ for 16h after 3h at room temperature. The stirring was stopped, and a saturated potassium carbonate solution was added thereto and stirred for 30mm. Then, water was added to dilute (20 mL), ethyl acetate was extracted (20 mL × 3), the organic phases were combined, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to obtain 240mg of a pale red solid with a yield of 24.83%.
LC-MS：(pos.ion)m/z：864.4[M+1] + ；
Step 4) (2S) -1- (4- ((3 '- (3- (2,7-diazaspiro [4.4] non-2-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
To a solution of (2S) -1- (4- ((3 '- (3- (7- (tert-butoxycarbonyl) -2,7-diazaspiro [4.4] nonan-2-yl) propoxy) -2-methyl- [1,1' biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (240mg, 0.31mmol) in DCM (25 mL) was added trifluoroacetic acid (1 mL) and reacted at room temperature for 3h. The stirring was stopped and TLC was used for detection. The solvent was concentrated, and preparative isolation and purification gave 45mg of a pale yellow solid product with a yield of 18.85%.
LC-MS：(pos.ion)m/z：764.5[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.00(s，1H)，8.97(s，1H)，8.43(s，1H)，7.46(d，J＝10.0Hz，2H)，7.34(t，J＝7.9Hz，1H)，7.25(t，J＝7.4Hz，1H)，7.20(d，J＝7.3Hz，1H)，7.08(s，1H)，6.91(d，J＝8.2Hz，1H)，6.85(d，J＝7.5Hz，1H)，6.78(s，1H)，5.30(s，2H)，5.27(s，2H)，4.03(t，J＝6.2Hz，2H)，3.72(d，J＝13.8Hz，2H)，3.50(d，J＝13.3Hz，2H)，3.07(s，2H)，2.96(s，1H)，2.23(s，3H)，2.14(s，1H)，1.99(s，1H)，1.88-1.79(m，3H)，1.73(dd，J＝13.5，7.0Hz，6H)，1.44(s，3H)，1.24(s，7H).
Example 27N- (2- ((5-chloro-4- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -2- (pyridin-3-ylmethoxy) benzyl) amino) ethyl) acetamide (Compound 27)
Step 1) 5-chloro-2-hydroxy-4- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (800mg, 1.588mmol) and 2-hydroxy-2,7-diazaspiro [3.5] nonane hydrochloride (423.2mg, 2.38mmol) in DMF (20 mL) under nitrogen protection was added potassium carbonate (658.4mg, 4.76mmol), sodium iodide (357mg, 2.38mmol) in that order, followed by heating to 70 ℃ for 16h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (80 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 437mg of yellow solid in 48.79% yield.
LC-MS：(pos.ion)m/z：564.2[M+1] + ；
Step 2) 5-chloro-4- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -pyridin-3-yl) methoxy) -2- (pyridin-3-ylmethoxy) benzaldehyde
To 5-chloro-2-hydroxy-4- ((3' - (3- (2-hydroxy-7-azaspiro [ 3.5))]Nonan-7-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) methoxy) benzaldehyde (330mg, 0.58mmol) and 3-bromomethylpyridinobromite (221.9mg, 0.87mmol) in DMF (20 mL) were added cesium carbonate (571.8mg, 1.75mmol) and sodium iodide (17.54mg, 0.111mmol), N in that order 2 Protecting, heating to 75 ℃ and reacting for 4h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (80 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 240mg of red oily liquid with a yield of 62.62%.
LC-MS：(pos.ion)m/z：655.1[M+1] + ；
Step 3) N- (2- ((5-chloro-4- ((3 ' - (3- (2-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -2- (pyridin-3-ylmethoxy) benzyl) amino) ethyl) acetamide
To 5-chloro-4- ((3' - (3- (2-hydroxy-7-azaspiro [3.5]]Nonan-7-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-pyridin-3-yl) methoxy) -2- (pyridin-3-ylmethoxy) benzaldehyde (150mg, 0.22mmol) and N- (2-aminoethyl) acetamide (35.08mg, 0.34mmol) in CH 3 Adding acetic acid into OH (10 mL) solution, adjusting the pH value to about 5, reacting at room temperature for 15min, adding sodium borocyanide (71.93mg, 1.14mmol) into the reaction system, and reacting at room temperature for 2h. Stirring was stopped, saturated potassium carbonate solution was added and stirred for 30min. Water dilution (50 mL), ethyl acetate extraction (80 mL × 3), organic phases combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 50mg of a white solid in 29.46% yield.
LC-MS：(pos.ion)m/z：741.4[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ8.70(s，1H)，8.55(s，1H)，7.89(d，J＝7.6Hz，1H)，7.80(s，1H)，7.45(dd，J＝17.6，6.3Hz，2H)，7.37(s，1H)，7.26(t，J＝7.3Hz，1H)，7.19(t，J＝7.6Hz，1H)，7.12(s，1H)，7.06(d，J＝7.3Hz，1H)，6.94(d，J＝8.0Hz，1H)，6.67(d，J＝7.5Hz，1H)，5.24(s，4H)，4.04(dd，J＝16.7，6.9Hz，3H)，3.63(s，2H)，3.11(d，J＝5.9Hz，2H)，2.42(s，2H)，2.27(d，J＝14.2Hz，4H)，2.09-1.96(m，6H)，1.87(d，J＝15.6Hz，3H)，1.82(s，3H)，1.77(s，3H)，1.53-1.42(m，6H).
Example 121N- (2- ((4- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] nonan-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzyl) amino) ethyl) acetamide (Compound 121)
Step 1) 4- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (1000mg, 1.95mmol) and 2-oxa-7-azaspiro [4.4] nonane (378.7mg, 2.97mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (411.5mg, 2.97mmol), followed by sodium iodide (446.3mg, 2.97mmol), followed by heating to 70 ℃ for 16h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 680mg of a reddish brown solid in 62.28% yield.
LC-MS：(pos.ion)m/z：550.1[M+1] - ；
Step 2) 4- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzaldehyde
To 4- ((3' - (3- (2-oxa-7-azaspiro [4.4 ])]Nonan-7-yl) propoxyRadix) -2,2 '-dimethyl- [1,1' -biphenyl]To a solution of (420mg, 0.76mmol) of-3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde and (231.7mg, 0.91mmol) of 3-bromomethylpyridinobromite (231.7mg, 0.91mmol) in DMF (20 mL) were added cesium carbonate (621.9mg, 1.9mmol) and sodium iodide (22.89mg, 0.15mmol), N 2 Protecting, heating to 75 ℃ and reacting for 4h. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (80 mL × 3), the organic phases combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 250mg of a yellow solid in 51.07% yield.
LC-MS：(pos.ion)m/z：641.1[M+1] + ；
Step 3) N- (2- ((4- ((3 ' - (3- (2-oxa-7-azaspiro [4.4] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzyl) amino) ethyl) acetamide
To 4- ((3' - (3- (2-oxa-7-azaspiro [4.4 ])]Nonan-7-yl) propoxy) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) methoxy) -5-chloro-2- (pyridin-3-ylmethoxy) benzaldehyde (150mg, 0.22mmol) and N- (2-aminoethyl) acetamide (35.08mg, 0.34mmol) in CH 3 Adding acetic acid into OH (10 mL) solution, adjusting the pH value to 5, reacting at room temperature for 15min, adding sodium borocyanide (71.93mg, 1.14mmol) into the reaction system, and reacting at room temperature for 2h. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Then, water was added to dilute (50 mL), ethyl acetate was extracted (80 mL × 3), the organic phases were combined, and the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to obtain 50mg of a white solid with a yield of 29.46%.
LC-MS：(pos.ion)m/z：727.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ8.70(s，1H)，8.55(s，1H)，7.89(d，J＝7.1Hz，1H)，7.80(s，1H)，7.51-7.41(m，2H)，7.37(s，1H)，7.26(t，J＝7.3Hz，1H)，7.19(t，J＝7.6Hz，1H)，7.12(s，1H)，7.06(d，J＝7.1Hz，1H)，6.95(d，J＝7.9Hz，1H)，6.67(d，J＝7.3Hz，1H)，5.24(s，4H)，4.04(d，J＝5.5Hz，2H)，3.69(s，2H)，3.63(s，2H)，3.55-3.49(m，2H)，3.41(d，J＝7.8Hz，2H)，3.11(d，J＝5.7Hz，2H)，2.56(s，2H)，2.37(d，J＝8.4Hz，1H)，2.04(s，3H)，1.90(s，3H)，1.82(s，5H)，1.77(s，4H)，1.74(d，J＝7.2Hz，3H).
Example 29N- (2- ((5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2- (pyridin-3-ylmethoxy) benzyl) amino) ethyl) acetamide (Compound 29)
Step 1) 5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2-hydroxybenzaldehyde
To a solution of 4- [ [3- [3- (3-bromopropoxy) -2-methyl-phenyl ] methoxy ] -5-chloro-2-hydroxy-benzaldehyde (800mg, 1.58mmol) and hexahydro-1H-furo [3,4-C ] pyrrole hydrochloride (356.4mg, 2.38mmol) in DMF (20 mL) under nitrogen protection was added sequentially potassium carbonate (658.4mg, 4.76mmol), sodium iodide (357mg, 2.38mmol) and then heated to 70 ℃ for 16H. Stirring was stopped, cooled to room temperature, diluted with water (50 mL), extracted with ethyl acetate (100 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 400mg of yellow oily liquid in 62.28% yield.
LC-MS：(pos.ion)m/z：536.3[M+1] - ；
Step 2) 5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2- (pyridin-3-ylmethoxy) benzaldehyde
To 5-chloro-4- ((2,2 '-dimethyl-3' - (3- (tetrahydro-1H-furo [3,4-c)]Pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl]-3-yl) methoxy) -2-hydroxybenzaldehyde (4)00mg, 0.74mmol) and 3-bromomethylpyridinium bromate (226.5mg, 0.89mmol) in DMF (20 mL), cesium carbonate (607.8mg, 1.86mmol) and sodium iodide (22.37mg, 0.14mmol), N 2 Protecting, heating to 75 ℃ and reacting for 4h. Stirring was stopped, cooling to room temperature was followed by dilution with water (50 mL), extraction with ethyl acetate (80 mL × 3), combination of the organic phases, washing of the organic phases with saturated brine (50 mL), drying over anhydrous sodium sulfate, filtration, concentration, and purification by column chromatography (DCM/MeOH =10/1,v/v) to give 300mg of red oily liquid with a yield of 51.07%.
LC-MS：(pos.ion)m/z：627.1[M+1] + ；
Step 3) N- (2- ((5-chloro-4- ((2,2 ' -dimethyl-3 ' - (3- (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl ] -3-yl) methoxy) -2- (pyridin-3-ylmethoxy) benzyl) amino) ethyl) acetamide
To 5-chloro-4- ((2,2 '-dimethyl-3' - (3- (tetrahydro-1H-furo [3,4-c)]Pyrrol-5 (3H) -yl) propoxy) - [1,1' -biphenyl]-3-yl) methoxy) -2- (pyridin-3-ylmethoxy) benzaldehyde (300mg, 0.4783mmol) and N- (2-aminoethyl) acetamide (73.28mg, 0.71mmol) in CH 3 Adding acetic acid into OH (10 mL) solution, adjusting the pH value to about 5, reacting at room temperature for 15min, adding sodium borocyanide (150.3 mg, 2.39mmol) into the reaction system, and continuing to react at room temperature for 2h. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (50 mL), ethyl acetate extraction (80 mL × 3), organic phases combined, organic phase washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, column chromatography purification (DCM/MeOH =15/1,v/v) afforded 110mg as a pale yellow solid in 32.24% yield.
LC-MS：(pos.ion)m/z：713.3[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ8.71(s，1H)，8.56(s，1H)，7.94-7.83(m，2H)，7.52-7.36(m，3H)，7.25(d，J＝6.9Hz，1H)，7.18(d，J＝7.3Hz，1H)，7.14(s，1H)，7.06(d，J＝6.8Hz，1H)，6.94(d，J＝7.8Hz，1H)，6.67(d，J＝7.0Hz，1H)，5.26(s，4H)，4.04(d，J＝5.1Hz，2H)，3.71(s，6H)，3.50(s，1H)，3.37(d，J＝5.8Hz，2H)，3.14(d，J＝5.0Hz，2H)，2.68(s，2H)，2.59(s，2H)，2.34(d，J＝7.5Hz，2H)，2.03(s，3H)，1.89(s，3H)，1.82(s，3H)，1.78(s，3H).
Example 30 (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (1-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 30)
Step 1) 5-chloro-2-hydroxy-4- ((3 ' - (3- (1-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzaldehyde
7-azaspiro [3.5] nonan-1-ol hydrochloride (0.4g, 2mmol) was dissolved in DMF (10.1 mL), and potassium carbonate (1.0g, 7.2mmol) was added thereto, followed by stirring at room temperature for 10min. 4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (0.5g, 1mmol), naI (0.2g, 1mmol) were added and stirred at 75 ℃ for 12h. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with EA (20 mL × 3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and separated by column chromatography (DCM/MeOH =10/1,v/v) to give 0.42g of a yellow solid in 80% yield.
LC-MS：(pos.ion)m/z：565.2[M+1] + ；
Step 2) 5- ((4-chloro-2-formyl-5- ((3 ' - (3- (1-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile
5- (chloromethyl) nicotinonitrile hydrochloride (0.17g, 0.90mmol) was dissolved in DMF (15.0 mL), and cesium carbonate (1.5g, 4.6 mmol) was added and stirred at room temperature for 10min. 5-chloro-2-hydroxy-4- ((3 ' - (3- (1-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzaldehyde (0.42g, 0.74mmol) and NaI (22.0mg, 0.147mmol) were added, and the mixture was stirred at 75 ℃ for 4 hours. Stirring was stopped, cooled to room temperature, diluted with water (15 mL), extracted with EA (15 mL × 5), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and isolated by column chromatography (DCM/MeOH =5/1,v/v) to give 0.37g of a yellow solid in 73% yield.
LC-MS：(pos.ion)m/z：681.1[M+1] + ；
Step 3) (2S) -1- (5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) -4- ((3 ' - (3- (1-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
5- ((4-chloro-2-formyl-5- ((3 ' - (3- (1-hydroxy-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) phenoxy) methyl) nicotinonitrile (0.37g, 0.54mmol) and D-pipecolic acid (0.14g, 1.1mmol) were dissolved in DMF (15.1 mL), acetic acid (0.2mL, 3mmol) was added, followed by heating to 60 ℃ and stirring for 1.5h, cooling to room temperature, sodium cyanoborohydride (0.17g, 2.7mmol) was slowly added, protected with nitrogen, and stirred for 12h at room temperature. Stirring was stopped, cooling to room temperature, adding saturated sodium bicarbonate solution (25 mL), stirring for 30min at room temperature, ea (25 mL × 3) extraction, combining the organic phases and washing with saturated brine (30 mL), drying over anhydrous sodium sulfate, suction filtration, washing, concentration under reduced pressure, column chromatography (DCM/MeOH =3/1,v/v) isolated 22.5mg of a pale yellow solid with a yield of 5%.
LC-MS：(pos.ion)m/z：794.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.01(d，J＝6.9Hz，2H)，8.46(s，1H)，7.49(d，J＝7.4Hz，1H)，7.43(s，1H)，7.29-7.21(m，2H)，7.12(d，J＝3.5Hz，1H)，7.07(d，J＝7.4Hz，1H)，6.98(d，J＝8.2Hz，1H)，6.71(d，J＝7.7Hz，1H)，5.34(s，2H)，5.27(s，2H)，4.14-4.04(m，2H)，3.81(d，J＝14.2Hz，2H)，3.65(d，J＝13.3Hz，2H)，3.51(s，1H)，3.19-3.13(m，4H)，2.95-2.87(m，2H)，2.38-2.27(m，2H)，2.22-2.16(m，3H)，2.12-2.07(m，1H)，2.03(s，3H)，1.84-1.71(m，9H)，1.55-1.43(m，4H)，1.43-1.33(m，2H).
EXAMPLE 31 (S) -1- (4- ((3 ' - (3- (2-amino-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate (Compound 31)
Step 1) (tert-butyl 7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-azaspiro [3.5] non-2-yl) carbamate
4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (0.5g, 1mmol), tert-butyl 7-azaspiro [3.5] non-2-ylaminecarboxylate (0.3g, 1mmol) were dissolved in DMF (10.1 mL), potassium carbonate (0.8g, 6 mmol) was added, naI (0.2g, 1mmol) was added, and the temperature was raised to 75 ℃ for 12h. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with EA (20 mL × 3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and separated by column chromatography (DCM/MeOH =10/1,v/v) to give 0.6g of red oil in 90% yield.
LC-MS：(pos.ion)m/z：664.2[M+1] + ；
Step 2) tert-butyl 7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-azaspiro [3.5] non-2-yl) carbamate
5- (chloromethyl) nicotinonitrile hydrochloride (0.2g, 1mmol) was dissolved in DMF (20.1 mL), and cesium carbonate (2.0 g, 6.1mmol) was added and stirred at room temperature for 10min. (7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-aza-spiro [3.5] non-2-yl) carbamic acid tert-butyl ester (0.6 g, 0.9mmol) and NaI (30.0mg, 0.200mmol) were added, warmed to 75 ℃ and stirred for 4h. Stirring was stopped, cooled to room temperature, diluted with water (15 mL), extracted with EA (15 mL × 5), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and separated by column chromatography (DCM/MeOH =10/1,v/v) to give 0.6g of a red solid with a yield of 90%.
LC-MS：(pos.ion)m/z：780.1[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (2- ((tert-butoxycarbonyl) amino) -7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
Tert-butyl (7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-aza-spiro [3.5] non-2-yl) carbamate (0.6 g, 0.8mmol) and D-pipecolic acid (0.2g, 2mmol) were dissolved in DMF (15.0 mL), acetic acid (0.2mL, 3mmol) was added, heating to 60 ℃ and stirring for 1.0h, cooling to room temperature, sodium cyanoborohydride (0.2g, 3.0mmol) was slowly added, nitrogen blanketed, and stirring at room temperature for 12h. Stirring was stopped, cooled to room temperature, added with saturated sodium bicarbonate solution (25 mL), stirred at room temperature for 30min, extracted with ea (25 mL × 3), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and column chromatographed (DCM/MeOH =3/1,v/v) to give 0.13g of a yellow solid with a yield of 20%.
LC-MS：(pos.ion)m/z：446.70[0.5(M+1)] + ；
Step 4) (S) -1- (4- ((3 ' - (3- (2-amino-7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate
(S) -1- (4- ((3 ' - (3- (2- ((tert-butoxycarbonyl) amino) -7-azaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (0.13g, 0.15mmol) was dissolved in DCM (25.1 mL), TFA (2.0 mL) was added and stirred at room temperature for 4h. The system was concentrated under reduced pressure and sent to preparative isolation to give 32.7mg of a pale yellow solid with a yield of 25%.
LC-MS：(pos.ion)m/z：396.7[0.5(M+1)] + ；
1 H NMR(600MHz，d 6 -DMSO)δ9.66(s，1H)，9.04(d，J＝14.6Hz，2H)，8.48(s，1H)，8.10(s，2H)，7.53(s，1H)，7.52-7.48(m，1H)，7.30-7.28(m，1H)，7.26-7.22(m，2H)，7.08(d，J＝7.5Hz，1H)，6.98(d，J＝8.3Hz，1H)，6.71(d，J＝7.5Hz，1H)，5.45-5.36(m，2H)，5.36-5.30(m，2H)，4.31-4.26(m，2H)，4.15-4.05(m，2H)，3.75-3.67(m，2H)，3.25(s，4H)，3.03-2.92(m，2H)，2.89-2.82(m，2H)，2.37-2.28(m，1H)，2.24-2.16(m，2H)，2.12-2.08(m，2H)，2.04(s，3H)，2.01-1.93(m，2H)，1.92-1.82(m，4H)，1.82-1.59(m，6H)，1.49(s，2H).
Example 32 (S) -1- (4- ((3 ' - (3- (2,7-diazaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate (Compound 32)
Step 1) tert-butyl 7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [3.5] nonane-2-carboxylate
4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] 3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (0.5g, 1mmol), 2,7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester (0.3g, 1mmol) were dissolved in DMF (10.1 mL), potassium carbonate (0.8g, 6mmol) was added, naI (0.2g, 1mmol) was added, and the mixture was stirred at 75 ℃ for 12h. Stirring was stopped, cooled to room temperature, diluted with water (30 mL), extracted with EA (20 mL × 3), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and separated by column chromatography (DCM/MeOH =100/1,v/v) to give 0.533g of a red solid in 80% yield.
LC-MS：(pos.ion)m/z：650.2[M+1] + ；
Step 2) 7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester
5- (chloromethyl) nicotinonitrile hydrochloride (0.186g, 0.984mmol) was dissolved in DMF (10.1 mL), cesium carbonate (1.61g, 4.94mmol) was added, and the mixture was stirred at room temperature for 10min. 7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester (0.533g, 0.821mmol) and NaI (24.6mg, 0.164mmol) were added and stirred at 75 ℃ for 4h. Stirring was stopped, cooled to room temperature, diluted with water (15 mL), extracted with EA (15 mL × 5), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and isolated by column chromatography (DCM/MeOH =10/1,v/v) to give 0.6g of a yellow solid with a yield of 95%.
LC-MS：(pos.ion)m/z：766.3[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (2- (tert-butoxycarbonyl) -2,7-diazaspiro [3.5] nonan-7-yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester (0.6 g, 0.8mmol) and D-pipecolic acid (0.2g, 2mmol) were dissolved in DMF (15.0 mL), acetic acid (0.2mL, 3mmol) was added, heating to 60 ℃ and stirring was continued for 1.5h, followed by cooling to room temperature, slow addition of sodium cyanoborohydride (0.25g, 4.0mmol), nitrogen blanketing and stirring at room temperature for 12h. Stirring was stopped, saturated sodium bicarbonate solution (25 mL) was added and stirred for 30min at room temperature, ea (25 mL × 3) was extracted, the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered under suction, washed, concentrated under reduced pressure, and column chromatographed (DCM/MeOH =3/1,v/v) to give 0.44g of yellow solid in 60% yield.
LC-MS：(pos.ion)m/z：879.3[M+1] + ；
Step 4) (S) -1- (4- ((3 ' - (3- (2,7-diazaspiro [3.5] non-7-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate
(S) -1- (4- ((3 ' - (3- (2- (tert-butoxycarbonyl) -2,7-diazaspiro [3.5] nonan-7-yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (0.44g, 0.50mmol) was dissolved in DCM (25.1 mL), TFA (2.5 mL) was added and stirred at room temperature for 4h. The solvent was concentrated and sent to preparative isolation to give 66.0mg of a yellow solid in 15% yield.
LC-MS：(pos.ion)m/z：779.2[M+1] + ；
1 H NMR(600MHz，d 6 -DMSO)δ10.14(s，1H)，9.19(s，2H)，9.04(s，2H)，8.49(s，1H)，7.52(d，J＝29.4Hz，2H)，7.32-7.26(m，1H)，7.26-7.17(m，2H)，7.08(d，J＝6.4Hz，1H)，6.97(d，J＝7.4Hz，1H)，6.71(d，J＝6.8Hz，1H)，5.44-5.38(m，2H)，5.37-5.28(m，2H)，4.40-4.32(m，2H)，4.31-4.23(m，2H)，3.91-3.82(m，4H)，3.79-3.72(m，2H)，3.55-3.50(m，2H)，3.29-3.19(m，2H)，3.11-2.81(m，4H)，2.80-2.67(m，1H)，2.23-2.14(m，4H)，2.04(s，3H)，1.96-1.88(m，2H)，1.85(s，3H)，1.78-1.58(m，4H).
Example 33 (S) -1- (4- ((3 ' - (3- (2,8-diazaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 33)
Step 1) tert-butyl 8- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,8-diazaspiro [4.5] decane-2-carboxylate
2,7-diazaspiro [4.5] decane-2-carboxylic acid tert-butyl ester hydrochloride (0.3 g, 1mmol) was dissolved in DMF (10.1 mL), potassium carbonate (0.8g, 6mmol) was added, stirring was carried out at room temperature for 10min, 4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy ] -5-chloro-2-hydroxybenzaldehyde (0.5g, 1mmol) was added, stirring was carried out at room temperature for 12h, stirring was stopped, cooling was carried out to room temperature, water (30 mL) was added for dilution, EA (20 mL. Times.3) was extracted, the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, suction filtered, washed, concentrated under reduced pressure, and separated by silica gel column chromatography (DCM/MeOH =8/1,v/v) to give a pink solid in a yield of 70%.
LC-MS：(pos.ion)m/z：664.5[M+1] + ；
Step 2) tert-butyl 8- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,8-diazaspiro [4.5] decane-2-carboxylate
5- (chloromethyl) nicotinonitrile hydrochloride (0.15g, 0.79mmol) was dissolved in DMF (10.1 mL), and cesium carbonate (1.3g, 4.0 mmol) was added and stirred at room temperature for 10min. 8- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,8-diazaspiro [4.5] decane-2-carboxylic acid tert-butyl ester (0.43g, 0.65mmol) and NaI (19.0 mg, 0.127mmol) were added, warmed to 75 ℃ and stirred for 4h. Stirring was stopped, cooled to room temperature, diluted with water (15 mL), extracted with EA (15 mL × 5), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and chromatographed on silica gel (DCM/MeOH =5/1,v/v) to give 0.47g of a brown solid in 93% yield.
LC-MS：(pos.ion)m/z：780.3[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (2- (tert-butoxycarbonyl) -2,8-diazaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
Reacting 8- (3- ((3- ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2 '-dimethyl- [1,1' -biphenyl]-3-yl) oxy) propyl) -2,8-diazaspiro [4.5]Tert-butyl decane-2-carboxylate (0.47g, 0.60mmol) and D-pipecolic acid (0.16g, 1.2mmol) were dissolved in DMF (15.1 mL), acOH (0.2mL, 3mmol) was added, heating was carried out to 60 ℃ and stirring was carried out for 1.5h, cooling was carried out to room temperature, and NaBH was slowly added 3 CN (0.2g, 3.0mmol), nitrogen protection, room temperatureStirring for 12h. Stirring was stopped, cooling to room temperature, adding saturated sodium bicarbonate solution and stirring at room temperature for 30min, ea (25 mL × 3) extraction, washing with saturated brine (30 mL), drying over anhydrous sodium sulfate, suction filtration, washing, concentration under reduced pressure, and column chromatography (DCM/MeOH =5/1,v/v) to isolate 0.16g of a red solid with a yield of 30%.
LC-MS：(pos.ion)m/z：446.8[0.5(M+1)] + ；
Step 4) (S) -1- (4- ((3 ' - (3- (2,8-diazaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
(S) -1- (4- ((3 ' - (3- (2- (tert-butoxycarbonyl) -2,8-diazaspiro [4.5] decan-8-yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (0.16g, 0.18mmol) was dissolved in DCM (25.0 mL), TFA (2.0 mL) was added and stirred at room temperature for 4h. The solvent was concentrated, the organic solvent was concentrated after preparative separation, the aqueous solution was adjusted to pH 7-8 with saturated sodium carbonate, extracted sequentially with DCM (10 mL. Times.3) and EA (10 mL. Times.1), the organic phase was washed with water (20 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, and the organic phase was concentrated to give 22.8mg of a yellow solid in 16% yield.
LC-MS：(pos.ion)m/z：397.3[0.5(M+1)] + ；
1 H NMR(400MHz，d 6 -DMSO)δ8.98(d，J＝10.8Hz，2H)，8.43(s，1H)，7.53-7.41(m，2H)，7.26-7.21(m，1H)，7.19-7.15(m，1H)，7.07-7.03(m，2H)，6.98-6.87(m，1H)，6.67(d，J＝7.3Hz，1H)，5.35-5.19(m，4H)，4.81-4.74(m，1H)，4.03-3.98(m，2H)，3.51(s，1H)，3.08-3.03(m，2H)，2.86-2.79(m，4H)，2.43-2.36(m，2H)，2.34-2.29(m，2H)，2.25-2.21(m，2H)，2.11-1.92(m，5H)，1.86-1.73(m，5H)，1.70-1.65(m，2H)，1.63-1.55(m，2H)，1.55-1.34(m，8H).
Example 34 (S) -1- (4- ((3 ' - (3- (3,9-diazaspiro [5.5] undecan-3-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate (Compound 34)
Step 1) tert-butyl 9- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -3,9-diazaspiro [5.5] undecane-3-carboxylate
3,9-diazaspiro [5.5] undecane-9-carboxylic acid tert-butyl ester hydrochloride (0.3g, 1mmol) was dissolved in DMF (10.1 mL), potassium carbonate (0.8g, 6mmol) was added, and the mixture was stirred at room temperature for 10min. 4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy ] -5-chloro-2-hydroxybenzaldehyde (0.5g, 1mmol), naI (0.2g, 1mmol) were added, and the mixture was stirred at a temperature of 75 ℃ for 12 hours, the stirring was stopped, the mixture was cooled to room temperature, diluted with water (30 mL), EA (20 mL. Times.3) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, filtered, washed, concentrated under reduced pressure, and subjected to column chromatography (DCM/MeOH =8/1,v/v) to obtain 0.63g of a viscous yellow oil with a yield of 90%.
LC-MS：(pos.ion)m/z：678.0[M+1] + ；
Step 2) tert-butyl 9- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -3,9-diazaspiro [5.5] undecane-3-carboxylate
5- (chloromethyl) nicotinonitrile hydrochloride (0.21g, 1.1mmol) was dissolved in DMF (15.1 mL), and cesium carbonate (1.8g, 5.5mmol) was added and stirred at room temperature for 10min. 9- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -3,9-diazaspiro [5.5] undecane-3-carboxylic acid tert-butyl ester (0.63g, 0.93mmol) and NaI (28.0mg, 0.19mmol) were added and the mixture was allowed to warm to 75 deg.C and stirred for 4h. Stirring was stopped, cooled to room temperature, diluted with water (15 mL), extracted with EA (15 mL × 5), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and separated by column chromatography (DCM/MeOH =5/1,v/v) to give 0.63g of a red solid in 85% yield.
LC-MS：(pos.ion)m/z：397.2[0.5(M+1)] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (9- (tert-butoxycarbonyl) -3,9-diazaspiro [5.5] undecan-3-yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
9- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -3,9-diazaspiro [5.5] undecane-3-carboxylic acid tert-butyl ester (0.63g, 0.79mmol) and D-pipecolic acid (0.21g, 1.6mmol) were dissolved in DMF (15.1 mL), acetic acid (0.2mL, 3mmol) was added, heated to 60 ℃ and stirred for 1.0h, cooled to room temperature, sodium cyanoborohydride (0.25g, 4.0mmol) was slowly added, protected with nitrogen, and stirred for 12h at room temperature. The temperature is increased to 80 ℃ and the mixture is stirred for 3.0h. Stirring was stopped, cooling to room temperature, adding saturated sodium bicarbonate solution (25 mL), stirring for 30min at room temperature, ea (25 mL × 3) extraction, combining the organic phases and washing with saturated brine (30 mL), drying over anhydrous sodium sulfate, suction filtration, washing, concentrating under reduced pressure, and column chromatography (DCM/MeOH =5/1,v/v) isolated as a yellow solid 0.13g with 18% yield.
LC-MS：(pos.ion)m/z：454.0[0.5(M+1)] + ；
Step 4) (S) -1- (4- ((3 ' - (3- (3,9-diazaspiro [5.5] undecan-3-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate
(S) -1- (4- ((3 ' - (3- (9- (tert-butoxycarbonyl) -3,9-diazaspiro [5.5] undecan-3-yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (0.13g, 0.14mmol) was dissolved in DCM (25.0 mL), TFA (2.0 mL) was added and stirred at room temperature for 4h. The system was concentrated under reduced pressure and the preparation isolated to give 32.9mg of a pale yellow solid in 25% yield.
LC-MS：(pos.ion)m/z：404.4[0.5(M+1)] + ；
1 H NMR(600MHz，d 6 -DMSO)δ9.44(s，1H)，9.03(d，J＝15.5Hz，2H)，8.53(s，2H)，8.47(s，1H)，7.54-7.43(m，2H)，7.32-7.26(m，1H)，7.25-7.21(m，1H)，7.16(d，J＝4.4Hz，1H)，7.08-7.06(m，1H)，6.98(d，J＝8.4Hz，1H)，6.72(d，J＝7.5Hz，1H)，5.39-5.34(m，2H)，5.33-5.27(m，2H)，4.13-4.07(m，2H)，3.63-3.50(m，6H)，3.22-3.16(m，4H)，3.13-3.00(m，6H)，2.21-2.16(m，2H)，2.04(s，3H)，1.90-1.84(m，4H)，1.79-1.76(m，2H)，1.74-1.64(m，2H)，1.60-1.50(m，6H)，1.45-1.34(m，2H).
Example 35 (S) -1- (4- ((3 ' - (3- (2,9-diazaspiro [5.5] undecan-9-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate (Compound 35)
Step 1) tert-butyl 9- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,9-diazaspiro [5.5] undecane-2-carboxylate
4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy ] -5-chloro-2-hydroxybenzaldehyde (0.75g, 1.5mmol), 2,9-diazaspiro [5.5] undecane-4-carboxylic acid tert-butyl ester (0.45g, 1.8mmol) were dissolved in DMF (10.1 mL), potassium carbonate (1.2g, 8.7mmol) was added, naI (0.27g, 1.8mmol) was added, temperature was raised to 75 ℃ and stirring was continued for 12h, stirring was stopped, cooling was effected to room temperature, dilution with water (30 mL) was added, EA (20 mL. Times.3) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, suction filtration, washing, concentration under reduced pressure, separation (DCM) gave 0.8g of red oil with a yield of 80%.
LC-MS：(pos.ion)m/z：678.2[M+1] + ；
Step 2) tert-butyl 9- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,9-diazaspiro [5.5] undecane-2-carboxylate
5- (chloromethyl) nicotinonitrile hydrochloride (0.3 g, 2mmol) was dissolved in DMF (20.1 mL), cesium carbonate (2.0 g, 6.1mmol) was added, and stirring was carried out at room temperature for 10min. 9- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,9-diazaspiro [5.5] undecane-2-carboxylic acid tert-butyl ester (0.8g, 1mmol) and NaI (40.0mg, 0.267mmol) were added, and the mixture was heated to 75 ℃ and stirred for 4h. Stirring was stopped, cooled to room temperature, diluted with water (15 mL), extracted with EA (15 mL × 5), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and separated by column chromatography (DCM/MeOH =10/1,v/v) to give 0.6g of a pale red solid in 60% yield.
LC-MS：(pos.ion)m/z：794.2[M+1] + ；
Step 3) (S) -1- (4- ((3 ' - (3- (2- (tert-butoxycarbonyl) -2,9-diazaspiro [5.5] undecan-9-yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
Dissolving 9- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,9-diazaspiro [5.5] undecane-2-carboxylic acid tert-butyl ester (0.6 g, 0.8mmol) and D-pipecolic acid (0.2g, 2mmol) in DMF (15.0 mL), adding acetic acid (0.2mL, 3mmol), heating to 60 deg.C, stirring for 1.5h, cooling to room temperature, slowly adding sodium cyanoborohydride (0.2g, 3mmol), stirring for 12h under nitrogen protection, heating to 80 deg.C, and stirring for 5h. Stirring was stopped, cooling to room temperature, addition of saturated sodium bicarbonate solution (25 mL) stirring 30min at room temperature, ea (25 mL × 3) extraction, combining the organic phases and washing with saturated brine (30 mL), drying over anhydrous sodium sulfate, suction filtration, washing, concentration under reduced pressure, column chromatography (DCM/MeOH =4/1,v/v) isolated yellow solid 0.38g, yield 60%.
LC-MS：(pos.ion)m/z：907.3[M+1] + ；
Step 4) (S) -1- (4- ((3 ' - (3- (2,9-diazaspiro [5.5] undec-9-yl) propoxy) -2,2' -dimethyl- [1,1' -biphenyl ] methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate
(S) -1- (4- ((3 ' - (3- (2- (tert-butoxycarbonyl) -2,9-diazaspiro [5.5] undecan-9-yl) propoxy) -2,2' -dimethyl [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (0.38g, 0.42mmol) was dissolved in DCM (25.1 mL), TFA (2.5 mL) was added and stirred at room temperature for 4h. The system was concentrated under reduced pressure and sent to preparative isolation to give 47.6mg of a yellow solid in 12% yield.
LC-MS：(pos.ion)m/z：807.3[M+1] + ；
1 H NMR(600MHz，d 6 -DMSO)δ9.88(s，1H)，9.03(d，J＝7.7Hz，2H)，8.94(s，1H)，8.83(s，1H)，8.48(s，1H)，7.54(s，1H)，7.52-7.46(m，1H)，7.29(t，J＝6.6Hz，1H)，7.26-7.18(m，2H)，7.08(d，J＝7.4Hz，1H)，6.98(d，J＝8.1Hz，1H)，6.71(d，J＝7.4Hz，1H)，5.44-5.37(m，2H)，5.36-5.27(m，2H)，4.37-4.33(m，1H)，4.28-4.24(m，1H)，4.12-4.07(m，2H)，4.05-3.98(m，2H)，3.45-3.38(m，4H)，3.35-3.26(m，4H)，3.25-3.17(m，2H)，3.11(d，J＝4.0Hz，2H)，3.00(s，2H)，2.90-2.83(m，1H)，2.25-2.20(m，2H)，2.16-2.08(m，1H)，2.04(s，3H)，1.86(s，3H)，1.75-1.68(m，6H)，1.64-1.57(m，2H)，1.45(s，1H).
EXAMPLE 36 (S) -1- (4- ((3 '- (3- (2-amino-7-azaspiro [3.5] non-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate (Compound 36)
Step 1) [ 2-methyl-3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl ] methanol
PdCl 2 (dppf) (7.28g, 9.95mmol), potassium acetate (29.3g, 299mmol) and pinacol diboron (37.9g, 149mmol) were dissolved in 1,4-dioxane (200.1 mL), and (3-bromo-2-methyl-phenyl) methanol (20.1g, 100mmol) was added and stirred at 90 ℃ for 12h. Stirring was stopped, cooled to room temperature, and subjected to direct column chromatography (Hexane/EA =5/1,v/v) gave 23.9g of a pale green solid in 96% yield.
LC-MS：(pos.ion)m/z：231.1[M-18+1] + ；
Step 2) 1-bromo-3- (3-bromopropoxy) benzene
3-bromophenol (5.1g, 29mmol) was dissolved in acetone (60.0 mL), potassium carbonate (12.0g, 86.8mmol) was added, 1,3-dibromopropane (7.3mL, 72mmol) was then added, nitrogen was added, and the mixture was heated to 60 ℃ under reflux for 12h. Stirring was stopped, the mixture was cooled to room temperature, potassium carbonate was removed by suction filtration and washed with DCM, and 8.3g of a pale yellow oil was isolated by direct column chromatography (Hexane) under reduced pressure with a yield of 96%.
Step 3) (3 '- (3-bromopropoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methanol
Reacting [ 2-methyl-3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl]Methanol (4.4g, 18mmol), 1-bromo-3- (3-bromopropoxy) benzene (5.0g, 17mmol) was dissolved in THF (160.2 mL) and H 2 O (40.3 mL), potassium phosphate (9.0 g, 42mmol) was added under nitrogen, the mixture was stirred at room temperature for 20min, and Pd (dppf) Cl was added 2 (0.62g, 0.85mmol), nitrogen protection, heating and refluxing for 12h. Stirring was stopped, water (60 mL) was added for dilution, EA (100 mL × 3) extraction was performed, the organic phases were combined and dried over anhydrous sodium sulfate, suction filtration, washing, concentration under reduced pressure, and column chromatography (PE/EA =5/1,v/v) gave 3.6g of brown oil in 63% yield.
LC-MS：(pos.ion)m/z：318.2[M-18+1] + ；
Step 4) 4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy ] -5-chloro-2-hydroxybenzaldehyde
(3 '- (3-Bromopropoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methanol (2.0g, 6.0mmol) and 5-chloro-2,4-dihydroxybenzaldehyde (1.1g, 6.4mmol) were dissolved in THF (30.1 mL) at 0 deg.C, triphenylphosphine (2.3g, 8.8mmol) was added, nitrogen was added, DIAD (1.8mL, 9.1mmol) was slowly injected, and the mixture was stirred at room temperature for 24h. Stirring was stopped, water (30 mL) was added for dilution, EA (30 mL × 3) was extracted, the organic phases were combined and dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and column chromatographed (PE/EA =10/1,v/v) to give 1.9g of a yellow solid with a yield of 65%.
Step 5) tert-butyl 7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-azaspiro [3.5] non-2-yl) carbamate
4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy ] -5-chloro-2-hydroxybenzaldehyde (0.5g, 1mmol), tert-butyl 7-azaspiro [3.5] non-2-ylcarbamate (0.3g, 1mmol) were dissolved in DMF (10.1 mL), potassium carbonate (0.8g, 6mmol) was added, naI (0.2g, 1mmol) was added, the mixture was heated to 75 ℃ and stirred for 10 hours, stirring was stopped, cooling to room temperature, water (30 mL) was added for dilution, EA (20 mL. Times.3) was extracted, the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, suction filtration, washing, concentration under reduced pressure, column chromatography (Hexane/EA =1/3,v/v) gave 0.33g of a red solid with a yield of 50%.
LC-MS：(pos.ion)m/z：650.2[M+1] + ；
Step 6) tert-butyl 7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-aza-spiro [3.5] non-2-yl) carbamate
5- (chloromethyl) nicotinonitrile hydrochloride (0.12g, 0.63mmol) was dissolved in DMF (10.1 mL), and cesium carbonate (0.99g, 3.0 mmol) was added and stirred at room temperature for 10min. (7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-aza-spiro [3.5] non-2-yl) carbamic acid tert-butyl ester (0.33g, 0.51mmol) and NaI (15.0 mg,0.10 mmol) were added and the mixture was stirred at 75 ℃ for 4h. Stirring was stopped, cooled to room temperature, diluted with water (15 mL), extracted with EA (15 mL × 5), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and isolated by column chromatography (DCM/MeOH =90/1,v/v) to give 0.3g of a yellow solid in 80% yield.
LC-MS：(pos.ion)m/z：766.2[M+1] + ；
Step 7) (S) -1- (4- ((3 '- (3- (2- ((tert-butoxycarbonyl) amino) -7-azaspiro [3.5] non-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
Tert-butyl (7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -7-aza-spiro [3.5] non-2-yl) carbamate (0.3g, 0.4mmol) and D-pipecolic acid (0.1g, 0.8mmol) were dissolved in DMF (15.0 mL), acetic acid (0.2ml, 3mmol) was added, heating to 60 ℃ and stirring for 1.5h, cooling to room temperature, sodium cyanoborohydride (0.12g, 1.9mmol) was slowly added, nitrogen gas was used for protection, stirring at room temperature for 12h, stirring was stopped, cooling to room temperature, saturated sodium bicarbonate solution (25 mL) was added, stirring for 30min, ea (25 mL × 3) was extracted, the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, column chromatography, washing under reduced pressure, washing (DCM = 323 g, yellow solid separation yield: 0.1 g/1.1 g/v).
LC-MS：(pos.ion)m/z：440.3[0.5(M+1)] + ；
Step 8) (S) -1- (4- ((3 '- (3- (2-amino-7-azaspiro [3.5] non-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate
(S) -1- (4- ((3 '- (3- (2- ((tert-butoxycarbonyl) amino) -7-azaspiro [3.5] non-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (0.1g, 0.1mmol) was dissolved in DCM (25.1 mL), TFA (2.0 mL) was added, and stirring was continued at room temperature for 4h. The system was concentrated under reduced pressure and the preparation isolated to give 41.3mg of a white viscous solid in 40% yield.
LC-MS：(pos.ion)m/z：779.2[M+1] + ；
1 H NMR(600MHz，d 6 -DMSO)δ9.59(s，1H)，9.04(d，J＝12.3Hz，2H)，8.47(s，1H)，8.10(s，2H)，7.54(s，1H)，7.50(d，J＝6.2Hz，1H)，7.43-7.35(m，1H)，7.34-7.27(m，1H)，7.23(d，J＝7.1Hz，2H)，6.97(d，J＝6.6Hz，1H)，6.90(d，J＝6.4Hz，1H)，6.86(s，1H)，5.44-5.38(m，2H)，5.36-5.25(m，2H)，4.38-4.32(m，1H)，4.31-4.20(m，1H)，4.13-4.07(s，2H)，3.47-3.41(m，4H)，3.25-3.19(m，2H)，3.02-2.92(m，2H)，2.91-2.80(m，2H)，2.35-2.28(m，1H)，2.25(s，3H)，2.18-2.06(m，4H)，2.03-1.93(m，2H)，1.89-1.85(m，1H)，1.85-1.60(m，6H)，1.60-1.38(m，2H).
EXAMPLE 37 (S) -1- (4- ((3 '- (3- (2,7-diazaspiro [3.5] non-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate (Compound 37)
Step 1) 7- (3- ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2 '-methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester
4- ((3 ' - (3-bromopropoxy) -2,2' -dimethyl- [1,1' -biphenyl ] -3-yl) methoxy ] -5-chloro-2-hydroxybenzaldehyde (0.5g, 1mmol), 2,7-diazaspiro [3.5] non-2-carboxylic acid tert-butyl ester (0.3g, 1mmol) was dissolved in DMF (10.1 mL), potassium carbonate (0.8g, 6mmol) was added, naI (0.2g, 1mmol) was added, stirring was stopped at 75 ℃ for 12h, stirring was stopped, cooling was performed to room temperature, dilution with water (30 mL) was added, EA (20 mL. Times.3) was extracted, the organic phase was combined and washed with saturated brine (30 mL), anhydrous sodium sulfate was dried, suction filtered, washed, concentrated under reduced pressure, and isolated (Hexane/EA =1/3,v/v) to give a red solid in 60% yield.
LC-MS：(pos.ion)m/z：636.2[M+1] + ；
Step 2) 7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [3.5] non-2-carboxylic acid tert-butyl ester
5- (chloromethyl) nicotinonitrile hydrochloride (0.14g, 0.74mmol) was dissolved in DMF (10.1 mL), cesium carbonate (1.2g, 3.7mmol) was added, and stirring was carried out at room temperature for 10min. 7- (3- ((3 ' - ((2-chloro-4-formyl-5-hydroxyphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester (0.4g, 0.6 mmol) and NaI (19.0mg, 0.13mmol) were added and the mixture was allowed to warm to 75 ℃ and stirred for 4h. Stirring was stopped, cooled to room temperature, diluted with water (15 mL), extracted with EA (15 mL × 5), the organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered with suction, washed, concentrated under reduced pressure, and isolated by column chromatography (DCM/MeOH =10/1,v/v) to give 0.36g of a yellow solid in 80% yield.
LC-MS：(pos.ion)m/z：752.2[M+1] + ；
Step 3) (S) -1- (4- ((3 '- (3- (2- (tert-butoxycarbonyl) -2,7-diazaspiro [3.5] nonan-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
Dissolving 7- (3- ((3 ' - ((2-chloro-5- ((5-cyanopyridin-3-yl) methoxy) -4-formylphenoxy) methyl) -2' -methyl- [1,1' -biphenyl ] -3-yl) oxy) propyl) -2,7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester (0.36g, 0.48mmol) and D-pipecolic acid (0.12g, 0.93mmol) in DMF (15.0 mL), adding acetic acid (0.2mL, 3mmol), heating to 60 deg.C, stirring for 1.5h, cooling to room temperature, slowly adding sodium cyanoborohydride (0.15g, 2.4mmol), nitrogen protecting, and stirring at room temperature for 12h. Stirring was stopped, cooling was allowed to room temperature, saturated sodium bicarbonate solution (25 mL) was added and stirring was allowed to proceed for 30min at room temperature, ea (25 mL × 3) extraction, organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, washed, concentrated under reduced pressure, and column chromatography (DCM/MeOH =3/1,v/v) was performed to give 94.2mg of a yellow solid with a yield of 23%.
LC-MS：(pos.ion)m/z：432.8[0.5(M+1)] + ；
Step 4) (S) -1- (4- ((3 '- (3- (2,7-diazaspiro [3.5] non-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid trifluoroacetate
(S) -1- (4- ((3 '- (3- (2- (tert-butoxycarbonyl) -2,7-diazaspiro [3.5] non-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (94.2mg, 0.109mmol) was dissolved in DCM (25.1 mL), TFA (2.0 mL) was added and stirred at room temperature for 4h. The system was concentrated under reduced pressure and sent to preparative isolation to give 34.4mg of a yellow viscous solid in 36% yield.
LC-MS：(pos.ion)m/z：765.2[M+1] + ；
1 H NMR(600MHz，d 6 -DMSO)δ9.93(s，1H)，9.19-8.88(m，4H)，8.48(s，1H)，7.59-7.46(m，2H)，7.42-7.35(m，1H)，7.34-7.26(m，1H)，7.26-7.16(m，2H)，6.96(d，J＝6.9Hz，1H)，6.90(d，J＝6.3Hz，1H)，6.85(s，1H)，5.44-5.38(m，2H)，5.36-5.25(m，2H)，4.40-4.30(m，1H)，4.30-4.19(m，1H)，4.10(s，2H)，3.85(s，2H)，3.76(s，2H)，3.44-3.40(m，2H)，3.21(s，2H)，3.04-2.83(m，4H)，2.25(s，3H)，2.20(d，J＝13.6Hz，2H)，2.15(s，3H)，1.95-1.84(m，2H)，1.80-1.57(m，4H)，1.57-1.38(m，2H).
Example 67 (2S) -1- (4- ((3 '- (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid (Compound 67)
Step 1) 4- ((3 '- (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde
To a solution of 4- ((3 '- (3-bromopropoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2-hydroxybenzaldehyde (740mg, 1.51mmol) and 2-oxa-7-azaspiro [4.5] decane hydrochloride (402.6mg, 2.27mmol) in DMF (20 mL) under nitrogen protection were added potassium carbonate (522mg, 3.77mmol) and NaI (339.7mg, 2.26mmol) in this order, and the mixture was heated to 70 ℃ for 16 hours. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 370mg of yellow oily liquid, 44.52% yield.
LC-MS：(pos.ion)m/z：550.1[M+1] + ；
Step 2) 5- ((5- ((3 '- (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile
Mixing 4- ((3' - (3- (2-oxa-7-azaspiro [ 4.5))]Decan-7-yl) propoxy) -2-methyl- [1,1' -biphenyl]To a solution of (370mg, 067mmol) of (370mg) 3-chloro-2-hydroxybenzaldehyde and (148.8mg, 0.81mmol) of 5-chloromethyl-3-cyanopyridine hydrochloride in DMF (20 mL) were added cesium carbonate (534.3mg, 1.68mmol) and sodium iodide (19.66mg, 0.13mmol), N.sub. 2 And (4) protecting, and reacting for 4h at 75 ℃. Stirring was stopped, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL × 3), the organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =10/1,v/v) to give 400mg of yellow oily liquid in 89.27% yield.
LC-MS：(pos.ion)m/z：666.2[M+1] + ；
Step 3) (2S) -1- (4- ((3 '- (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -5-chloro-2- ((5-cyanopyridin-3-yl) methoxy) benzyl) piperidine-2-carboxylic acid
Adding acetic acid into a DMF (10 mL) solution of 5- ((5- ((3 '- (3- (2-oxa-7-azaspiro [4.5] decan-7-yl) propoxy) -2-methyl- [1,1' -biphenyl ] -3-yl) methoxy) -4-chloro-2-formylphenoxy) methyl) nicotinonitrile (400mg, 0.6 mmol) and D-pipecolic acid (155.1mg, 1.2 mmol), adjusting the pH to about 5, reacting at 60 ℃ for 1h, cooling to room temperature, adding sodium borocyanide (188.7mg, 3mmol) into the reaction system, reacting at room temperature for 3h, and reacting at 80 ℃ for 16h. Stopping stirring, adding saturated potassium carbonate solution, and stirring for 30min. Water dilution (20 mL), ethyl acetate extraction (20 mL × 3), combined organic phases, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM/MeOH =4/1,v/v) to give 92mg of light yellow solid in 19.66% yield.
LC-MS：(pos.ion)m/z：779.2[M+1] + ；
1 H NMR(400MHz，d 6 -DMSO)δ9.04-8.97(m，2H)，8.45(s，1H)，7.49(d，J＝7.2Hz，1H)，7.42(s，1H)，7.35(t，J＝7.9Hz，1H)，7.28(t，J＝7.5Hz，1H)，7.21(d，J＝7.2Hz，1H)，7.12(s，1H)，6.93(d，J＝8.3Hz，1H)，6.88-6.81(m，2H)，5.34(s，2H)，5.26(s，2H)，4.05(t，J＝6.2Hz，2H)，3.79(d，J＝13.9Hz，1H)，3.66(dd，J＝18.6，11.5Hz，4H)，3.55(d，J＝8.1Hz，1H)，3.51(s，1H)，3.32(d，J＝8.5Hz，2H)，3.15(d，J＝4.3Hz，1H)，2.34-2.26(m，2H)，2.24(s，4H)，1.90(d，J＝5.6Hz，2H)，1.79(s，1H)，1.75-1.67(m，2H)，1.56-1.31(m，10H)，1.23(s，1H).
In vitro antitumor Activity assay of Compounds of the invention
The in vitro enzymology level detection method adopts PD-1/PD-L1 binding assay kit of Cisbio company
1. The experimental principle is as follows:
PD-1 protein has HIS label, PD-L1 ligand of PD-1 has hFc label, and anti-hFc antibody marked by Eu and anti-HIS antibody marked by XL665 are combined with two label proteins respectively. After laser excitation, energy can be transferred from the donor Eu to the acceptor XL665, so that XL665 emits light. After the inhibitor is added, the combination of PD-1 and ligand PD-L1 is blocked, so that Eu and XL665 are far away, energy cannot be transferred, and XL665 does not emit light.
2. The experimental method comprises the following steps:
PD1/PD-L1 binding assays were performed using the HTRF assay kit from Cisbio Bioassays (Cat. No. 64ICP01 PEG), the specific protocol being described with reference to the kit instructions. Briefly described, compounds were dissolved in DMSO solutions at 10mM using 384 Kong Baise enzyme plates. The compound stock solution was first diluted 40-fold with a diluent, and then diluted 5-fold using a kit dilution buffer containing 2.5% dmso. 4ul of dilution or dilution of the target compound was added to each well, with a final DMSO concentration of 0.5%. PD1 and PD-L1 solutions were added at 3ul per well, respectively, and 10ul of detection antibody prepared according to the product instructions was added to each well after preincubation of the compound with PD1 and PD-L1 for 10 minutes. After incubating the plates at room temperature overnight, data were obtained by reading the plates in a pheasar FS microplate reader (BMG, germany). The HTRF signal was calculated as 10000 × (665/620 ratio)Rate). The calculated signal for each compound was fitted to a sigmoidal dose-response curve with variable slope, and IC was obtained by curve fitting (GraphPad Prism 7) 50 The value is obtained.
The experimental results are as follows:
TABLE 2 evaluation of the inhibitory Activity of the Compounds of the present invention on the PD-1/PD-L1 interaction at the molecular level
The experimental conclusion is that:
as can be seen from the table, the compounds of the present invention significantly inhibit the interaction of PD-1 with PD-L1 at the molecular level, and thus are useful for treating diseases associated with the interaction of PD-1/PD-L1.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (8)
1. A compound comprising a compound of one of the following or a stereoisomer, geometric isomer, tautomer, or pharmaceutically acceptable salt of a compound of one of the following:
2. a pharmaceutical composition comprising a compound of claim 1, and a pharmaceutically acceptable adjuvant, or a combination thereof.
3. Use of a compound of claim 1 or a pharmaceutical composition of claim 2 in the manufacture of a medicament for the prevention, treatment, or amelioration of a disease associated with the PD-1/PD-L1 signaling pathway in a patient.
4. The use of claim 3, wherein the disease associated with the PD-1/PD-L1 signaling pathway is cancer, an infectious disease, or an autoimmune disease.
5. The use according to claim 4, wherein the cancer is a disease in which there is unlimited proliferation of cells in an organ or a body tissue; the infectious diseases are bacterial infectious diseases, viral infectious diseases or fungal infectious diseases; the autoimmune disease is an organ-specific autoimmune disease or a systemic autoimmune disease.
6. The use of claim 5, wherein the disease of unlimited proliferation of cells in an organ or body tissue is bone cancer, head and neck cancer, pancreatic cancer, skin cancer, malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, hodgkin's disease, non-Hodgkin's lymphoma, carcinoma of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemia, solid tumor of the child, lymphocytic lymphomas, cancer of the bladder, cancer of the kidney or ureter, cancer of the renal pelvis, tumor of the Central Nervous System (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, or an environmentally induced cancer or a combination of the foregoing; wherein the chronic or acute leukemia comprises acute myelocytic leukemia, chronic myelocytic leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia.
7. The use according to claim 5, wherein the viral infectious disease is AIDS, hepatitis A, hepatitis B, hepatitis C, hepatitis D, herpes virus infection, papillomavirus infection, and influenza.
8. The use according to claim 5, wherein the organ-specific autoimmune disease is chronic lymphocytic thyroiditis, hyperthyroidism, insulin dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, goodpasture's syndrome, primary biliary cirrhosis, multiple sclerosis of the encephalomyelitis, and acute idiopathic polyneuritis; the systemic autoimmune disease is rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease and autoimmune hemolytic anemia.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2018102038124 | 2018-03-13 | ||
| CN201810203812 | 2018-03-13 | ||
| PCT/CN2019/077582 WO2019174533A1 (en) | 2018-03-13 | 2019-03-11 | Small molecule pd-1/pd-l1 inhibitor and use thereof in drugs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111788193A CN111788193A (en) | 2020-10-16 |
| CN111788193B true CN111788193B (en) | 2023-04-04 |
Family
ID=67907412
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201980016053.3A Active CN111788193B (en) | 2018-03-13 | 2019-03-11 | PD-1/PD-L1 small molecule inhibitor and application thereof in medicines |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN111788193B (en) |
| WO (1) | WO2019174533A1 (en) |
Families Citing this family (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109803651B (en) | 2016-06-27 | 2022-05-31 | 凯莫森特里克斯股份有限公司 | Immunomodulatory compounds |
| WO2018200571A1 (en) | 2017-04-25 | 2018-11-01 | Arbutus Biopharma Corporation | Substituted 2,3-dihydro-1h-indene analogs and methods using same |
| IL272258B (en) | 2017-07-28 | 2022-08-01 | Chemocentryx Inc | Immunomodulator compounds |
| KR20200056989A (en) | 2017-08-08 | 2020-05-25 | 케모센트릭스, 인크. | Macrocyclic immunomodulator |
| UA126458C2 (en) | 2018-02-13 | 2022-10-05 | Гіліад Сайєнсіз, Інк. | Pd-1/pd-l1 inhibitors |
| US10568874B2 (en) | 2018-02-22 | 2020-02-25 | Chemocentryx, Inc. | Indane-amines as PD-L1 antagonists |
| JP7242702B2 (en) | 2018-04-19 | 2023-03-20 | ギリアード サイエンシーズ， インコーポレイテッド | PD-1/PD-L1 inhibitor |
| TWI732245B (en) | 2018-07-13 | 2021-07-01 | 美商基利科學股份有限公司 | Pd-1/pd-l1 inhibitors |
| WO2020086556A1 (en) | 2018-10-24 | 2020-04-30 | Gilead Sciences, Inc. | Pd-1/pd-l1 inhibitors |
| CN111747927B (en) * | 2019-03-29 | 2023-08-18 | 广州丹康医药生物有限公司 | Compounds as immunomodulators and uses thereof |
| CA3139526A1 (en) | 2019-05-15 | 2020-11-19 | Chemocentryx, Inc. | Triaryl compounds for treatment of pd-l1 diseases |
| KR20220024701A (en) | 2019-06-20 | 2022-03-03 | 케모센트릭스, 인크. | Compounds for the treatment of PD-L1 disease |
| WO2021007386A1 (en) | 2019-07-10 | 2021-01-14 | Chemocentryx, Inc. | Indanes as pd-l1 inhibitors |
| US11713307B2 (en) | 2019-10-16 | 2023-08-01 | Chemocentryx, Inc. | Heteroaryl-biphenyl amides for the treatment of PD-L1 diseases |
| CA3152714A1 (en) | 2019-10-16 | 2021-04-22 | Pingchen Fan | Heteroaryl-biphenyl amines for the treatment of pd-l1 diseases |
| CN114276328B (en) * | 2020-09-28 | 2023-09-01 | 北京康辰药业股份有限公司 | Compound as small molecule immunosuppressant, preparation method and application thereof |
| CN113135895A (en) * | 2021-04-30 | 2021-07-20 | 中国药科大学 | Novel biphenyl derivative, preparation method and medical application thereof |
| CN114524768A (en) * | 2022-01-13 | 2022-05-24 | 黑龙江中医药大学 | Syringopicroside aglycone derivative with antibacterial activity and synthesis method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105705489A (en) * | 2013-09-04 | 2016-06-22 | 百时美施贵宝公司 | Compounds useful as immunomodulators |
| CN106536515A (en) * | 2014-04-14 | 2017-03-22 | 百时美施贵宝公司 | Compounds useful as immunomodulators |
| WO2017066227A1 (en) * | 2015-10-15 | 2017-04-20 | Bristol-Myers Squibb Company | Compounds useful as immunomodulators |
-
2019
- 2019-03-11 WO PCT/CN2019/077582 patent/WO2019174533A1/en active Application Filing
- 2019-03-11 CN CN201980016053.3A patent/CN111788193B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105705489A (en) * | 2013-09-04 | 2016-06-22 | 百时美施贵宝公司 | Compounds useful as immunomodulators |
| CN106536515A (en) * | 2014-04-14 | 2017-03-22 | 百时美施贵宝公司 | Compounds useful as immunomodulators |
| WO2017066227A1 (en) * | 2015-10-15 | 2017-04-20 | Bristol-Myers Squibb Company | Compounds useful as immunomodulators |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111788193A (en) | 2020-10-16 |
| WO2019174533A1 (en) | 2019-09-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111788193B (en) | PD-1/PD-L1 small molecule inhibitor and application thereof in medicines | |
| CN112166110B (en) | SHP2 phosphatase inhibitors and methods of use thereof | |
| US10517858B2 (en) | Thiazolecarboxamides and pyridinecarboxamide compounds useful as PIM kinase inhibitors | |
| CN108473498B (en) | Pyrazolo [1,5a ] pyrimidine derivatives as IRAK4 modulators | |
| US10000507B2 (en) | Furo- and thieno-pyridine carboxamide compounds useful as pim kinase inhibitors | |
| JP6631616B2 (en) | 2-Amino-pyrido [2,3-d] pyrimidin-7 (8H) -one derivatives as CDK inhibitors and uses thereof | |
| KR102485100B1 (en) | Substituted urea derivatives and pharmaceutical uses thereof | |
| CN107011348B (en) | Heteroaryl pyridone and aza-pyridone compounds as inhibitors of BTK activity | |
| US9822124B2 (en) | Bicyclic heteroaromatic carboxamide compounds useful as Pim kinase inhibitors | |
| KR20210068010A (en) | Carboxamide as a ubiquitin-specific protease inhibitor | |
| CN106336413B (en) | Compounds as JAK inhibitors and uses thereof | |
| CN112574236A (en) | RET inhibitor, pharmaceutical composition and application thereof | |
| CN112574235A (en) | RET inhibitor, pharmaceutical composition and application thereof | |
| CN112442050A (en) | RET inhibitor, pharmaceutical composition and application thereof | |
| CN109790160B (en) | Pyrido five-membered aromatic ring compound, preparation method and application thereof | |
| US20230183237A1 (en) | Competitive and noncompetitive inhibitors of the muscarinic acetylcholine receptor m5 | |
| CN113620944A (en) | Novel RET inhibitors, pharmaceutical compositions thereof and uses thereof | |
| CN111793077B (en) | PD-1/PD-L1 small molecule inhibitor and application thereof in medicines | |
| WO2024083111A1 (en) | Novel heterocyclic compounds | |
| WO2023131122A1 (en) | Fused ring-substituted six-membered heterocyclic compound, and preparation method therefor and use thereof | |
| TWI690533B (en) | Compounds as cdk small-molecule inhibitors and uses thereof | |
| CN113121463A (en) | Amide compound with bicyclic structure and application thereof in medicines | |
| WO2023020486A1 (en) | Rip1 kinase inhibitor compound and composition and use thereof | |
| CN115232109A (en) | PD-1/PD-L1 small molecule inhibitor and application thereof in medicines | |
| CN113121393A (en) | Aromatic amide compound and application thereof in medicines |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 523808 No.1, Gongye North Road, Songshanhu Park, Dongguan City, Guangdong Province Patentee after: Guangdong Dongyangguang Pharmaceutical Co.,Ltd. Address before: 523808 No.1, Gongye North Road, Songshanhu Park, Dongguan City, Guangdong Province Patentee before: SUNSHINE LAKE PHARMA Co.,Ltd. |
|
| CP01 | Change in the name or title of a patent holder |