CN114790186A - Fused ring compound as EP4 antagonist and preparation method and application thereof - Google Patents

Abstract

The invention provides a compound shown as a formula I, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof; the compound has effective antagonism on EP4, has good affinity with EP4 receptors, and can be used as an EP4 antagonist.

Description

Fused ring compound as EP4 antagonist and preparation method and application thereof

PRIORITY INFORMATION

The present application claims priority and benefit of a patent application having patent application number 202110098734.8 filed on the intellectual property office of china on 25.01.2021, and is incorporated herein by reference in its entirety.

Technical Field

The invention belongs to the field of medicines, and particularly relates to a fused ring compound serving as an EP4 antagonist, and a preparation method and application thereof.

Background

Prostaglandin E ₂ (Prostaglandin E ₂ ，PGE ₂ ) Is an endogenous organismActive lipids, PGE ₂ The activation of prostaglandin receptors causes a wide range of upstream and downstream dependent biological responses (Legler, d.f. et al, hit.j biochem. cell biol.2010,42, p.198-201), and is involved in the regulation of a variety of physiological and pathological processes including inflammation, pain, renal function, cardiovascular system, pulmonary function, and cancer. It is reported that PGE ₂ Is highly expressed in cancerous tissues of various cancers, and PGE has been confirmed ₂ Associated with the occurrence, growth and development of cancer and disease conditions in patients. It is generally accepted that PGE ₂ Are associated with activation of cell proliferation and cell death (apoptosis) and play an important role in the process of cancer cell proliferation, disease progression and cancer metastasis.

PGE ₂ Among the receptors of (a), there are 4 kinds of subtypes, EP1, EP2, EP3 and EP4, which are widely distributed in various tissues. Among these subtypes, PGE ₂ Intervention in inflammatory reactions (including immune inflammatory reactions), relaxation of smooth muscle, pain, differentiation of lymphocytes, hypertrophy and proliferation of vascular mesangial cells, secretion of gastrointestinal mucus, and the like via the EP4 receptor. Thus, it is believed that EP4 receptor antagonists hold promise as anti-inflammatory and/or analgesic therapies with PGE ₂ Diseases associated with the EP4 pathway, such as inflammatory diseases, diseases accompanied by various pains, and the like.

EP4 is the major receptor involved in arthritic pain in rodent models of rheumatoid arthritis and osteoarthritis (see e.g. j. pharmacol. exp. ther.,325,425(2008)) which upon activation leads to the accumulation of the intracellular signaling molecule cAMP. Expression of the EP4 receptor has been detected on the peripheral nerve endings of pain receptors, macrophages and neutrophils, and these cell types have been shown to be extremely important for endometriosis. Studies have reported that oral administration of EP4 antagonists can reduce proteinuria in type 2 diabetic mice and inhibit the progression of diabetic nephropathy. Another study reported activation of EP4 and PGE in the bladder mucosa ₂ The increased production of (b) may be a significant cause of overactive bladder caused by prostatitis, and intravesical injection of EP4 antagonists is effective in ameliorating overactive bladder after prostatitis. Thus, selective EP4 antagonists are useful for the treatment of arthritis, including arthritis pain and endometriosisLocation disease, diabetic nephropathy, overactive bladder. Existing remedies for arthritis are based on either traditional NSAIDs (non-steroidal anti-inflammatory drugs) or selective COX-2 inhibitors, which can produce cardiovascular and/or gastrointestinal side effects. Whereas selective EP4 antagonists are less likely to produce cardiovascular side effects.

PGE ₂ The continuous activation of EP receptors (produced in large numbers by tumor cells) in the tumor microenvironment (Ochs et al, J neurochem.2016,136, p.1142-1154; Zelenay, s.et al, Cell 2015,162, p.1257-1270) promotes the accumulation and enhances the activity of a variety of immunosuppressive cells, including tumor-associated macrophages of type 2 (TAMS), Treg cells, and myeloid-derived suppressor cells (MDSCs). One of the main features of the immunosuppressive tumor microenvironment is the presence of a large number of MDSCs and TAMs, which in turn are closely associated with low overall survival rates in patients with gastric, ovarian, breast, bladder, hepatocellular carcinoma (HCC), head and neck, and other types of cancer. In addition, PGE is reported ₂ Immune tolerance is induced by inhibiting the accumulation of antigen presenting Dendritic Cells (DCs) in tumors and by inhibiting the activation of tumor infiltrating DCs (Wang et al, Trends in Molecular Medicine 2016,22, p.1-3). All of these PGEs ₂ The mediated effects will collectively help tumor cells evade immune surveillance. PGE ₂ Plays an important role in promoting the occurrence and development of tumors. PGE is found in various malignant tumors including colon cancer, lung cancer, breast cancer, head and neck cancer ₂ And its related receptors EP2, EP4 have increased expression levels and are often closely related to poor prognosis (Bhoshan, N.et al.Lung Cancer 101, 88-91). Therefore, selective blockade of EP2 and EP4 signaling pathways can inhibit tumorigenesis from developing by altering the tumor microenvironment, modulating tumor immune cells.

The existing preclinical research data show that specific antagonists of EP2 and EP4 can prevent or inhibit the growth of tumors to different degrees in animal models of colon cancer, esophageal cancer, lung cancer, breast cancer and the like. PGE at entry to clinic ₂ Among the receptor drugs, the EP4 antagonist Grapiprant developed by the company pfeiri has been FDA approved for the treatment of arthritis in dogs, while entering clinical phase II research on anti-tumor in 2015It is used for treating various solid tumors such as prostate cancer, non-small cell lung cancer and breast cancer (De Vito, V.et al.J Pharm Biomed Anal 118, 251-258). EP4 antagonist E7046 developed by wei-defense corporation also developed a related study in phase I of clinic in 2015, and a clinical trial in phase Ib for treating rectal cancer in combination with radiotherapy or chemoradiotherapy was developed in 2017. Ono-4578, developed by Ono Pharmaceutical, conducted clinical phase I studies in 2017 on advanced or metastatic solid tumors and phase I/II clinical trials in 2018 on single or combined nivolumab treatment of advanced solid tumors.

The EP4 antagonists have been developed in the field of treatment of inflammatory diseases, pain, cancer, etc., but there is still a need for further development of novel drugs as an improvement or replacement of the current drugs.

Disclosure of Invention

The invention aims to provide a fused ring compound for an EP4 antagonist and a preparation method and application thereof.

In a first aspect of the present invention, there is provided a compound of formula I, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug thereof:

wherein ring A is absent or 7-10 membered bridged cycloalkyl, 6 membered oxygenated heterocycloalkyl;

and when ring A is 6-membered oxoheterocycloalkyl, the radical segment

Is composed of

R ₁ Is selected from C ₁ -C ₆ Alkyl, -CONH-R ₁₁ 、-NHCO-R ₁₂ ；R ₁₁ 、R ₁₂ Each independently is hydrogen or C ₁ -C ₆ An alkyl group; r ₁ 、R ₁₁ 、R ₁₂ C as described in (1) ₁ -C ₆ Alkyl is optionally substituted by one or more R _f Substitution; when the substituent is plural, the R _f The same or different;

R ₂ selected from halogen, cyano, C ₂ -C ₆ Alkenyl radical, C ₂ -C ₆ Alkynyl, -CO-R ₂₁ ；R ₂₁ Is amino, hydroxy, C ₁ -C ₆ An alkyl group; wherein said amino or hydroxy is optionally independently substituted by C ₁ -C ₆ Alkyl substitution; said C is ₂ -C ₆ Alkenyl radical, said C ₂ -C ₆ Alkynyl and said C ₁ -C ₆ Alkyl is optionally each independently substituted with one or more R _f Substitution; when the substituent is plural, the R _f The same or different;

R ₃ is C ₁ -C ₆ Carboxyl or C ₁ -C ₆ An amide group;

R ₄ selected from hydrogen, hydroxy, halogen, cyano, C ₁ -C ₄ Alkyl or C ₁ -C ₄ A haloalkyl group;

r mentioned above _f Selected from hydroxy, halogen or cyano.

In a preferred embodiment of the invention, the compound has the structure shown in formula II, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug:

in a preferred embodiment of the invention, ring A is absent or 7-membered bridged cycloalkyl, 6-membered oxoheterocycloalkyl, and when ring A is 6-membered oxoheterocycloalkyl, the radical fragment

Is composed of

Preferably, the group

Is composed of

When ring A is absent, a radical fragment

Is composed of

Preferably, ring A, when absent, is a fragment of a group

Is composed of

Preferably, when ring A is a 7-membered bridged cycloalkyl group, the moiety

Is composed of

More preferably, when ring A is a 7-membered bridged cycloalkyl group, the moiety

Is composed of

In a preferred embodiment of the present invention, R ₁ is-CONH-R ₁₁ or-NHCO-R ₁₂ (ii) a Wherein R is ₁₁ 、R ₁₂ Each independently is C ₁ -C ₆ An alkyl group; preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is C ₁ -C ₆ An alkyl group; preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is methyl, ethyl, propyl, isopropyl; more preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is propyl or isopropyl; preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is an isopropyl group.

In a preferred embodiment of the invention, R ₂ Is halogen, cyano; preferably, R ₂ Is cyano.

In a preferred embodiment of the invention, R ₃ Is C ₁ -C ₆ A carboxyl group; preferably, R ₃ Is butyric acid.

In a preferred embodiment of the invention, R ₄ Selected from hydrogen, hydroxy, halogen, cyano, methyl or halomethyl; preferably, R ₄ Is hydrogen.

In a preferred embodiment of the present invention, the compound has a structure represented by any one of the following, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof:

in a second aspect of the invention, there is provided an intermediate B-1, a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof:

wherein ring A is absent or 7-10 membered bridged cycloalkyl, 6 membered oxygenated heterocycloalkyl;

R ₁ is selected from C ₁ -C ₆ Alkyl, -CONH-R ₁₁ 、-NHCO-R ₁₂ ；R ₁₁ 、R ₁₂ Each independently is hydrogen or C ₁ -C ₆ An alkyl group; r is ₁ 、R ₁₁ 、R ₁₂ C as described in (1) ₁ -C ₆ Alkyl is optionally substituted by one or more R _f Substitution; when the substituent is plural, the R _f The same or different;

R ₄ selected from hydrogen, hydroxy, halogen, cyano, C ₁ -C ₄ Alkyl or C ₁ -C ₄ A haloalkyl group; preferably R ₄ Is hydrogen;

r mentioned above _f Selected from hydroxy, halogen or cyano.

In a preferred embodiment of the present invention, when ring A is a 6-membered oxoheterocycloalkyl group,

is composed of

In a preferred embodiment of the invention, ring A is absent or 7-membered bridged cycloalkyl, 6-membered oxoheterocycloalkyl, a moiety

Is composed of

R ₁ is-CONH-R ₁₁ or-NHCO-R ₁₂ Wherein R is ₁₁ 、R ₁₂ Each independently is C ₁ -C ₆ Alkyl radical, R ₄ Is hydrogen;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is C ₁ -C ₆ Alkyl radical, R ₄ Is hydrogen;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is methyl, ethyl, propyl, R ₄ Is hydrogen;

more preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is propyl, R ₄ Is hydrogen;

preferably, the first and second liquid crystal films are made of a polymer,

is composed of

Preferably, the group

Is composed of

Preferably, the propyl group is isopropyl; preferably, the radical is a fragment

Is composed of

In a preferred embodiment of the present invention, the intermediate B-1 has a structure represented by any one of the following, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof:

in a third aspect of the invention, there is provided an intermediate B-2, a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof:

wherein, the ring A is 7-10 membered bridged cycloalkyl and 6 membered oxygen-containing heterocycloalkyl;

R ₁ is selected from C ₁ -C ₆ Alkyl, -CONH-R ₁₁ 、-NHCO-R ₁₂ ；R ₁₁ 、R ₁₂ Each independently is hydrogen or C ₁ -C ₆ An alkyl group; r is ₁ 、R ₁₁ 、R ₁₂ C as described in (1) ₁ -C ₆ Alkyl is optionally substituted by one or more R _f Substitution; when the substituent is plural, the R _f The same or different;

R ₄ selected from hydrogen, hydroxy, halogen, cyano, C ₁ -C ₄ Alkyl or C ₁ -C ₄ A haloalkyl group; preferably, R ₄ Is hydrogen;

r mentioned above _f Selected from hydroxyl, halogen and cyano.

In a preferred embodiment of the invention, ring A is a 6-membered oxoheterocycloalkyl group,

is composed of

In a preferred embodiment of the invention, R ₁ is-CONH-R ₁₁ or-NHCO-R ₁₂ (ii) a Wherein R is ₁₁ 、R ₁₂ Each independently is C ₁ -C ₆ Alkyl radical, R ₄ Is hydrogen;

preferably, the radical is a fragment

Is composed of

R ₁ is-CONH-R ₁₁ ，R ₁₁ Is C ₁ -C ₆ Alkyl radical, R ₄ Is hydrogen;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Methyl, ethyl, propyl, isopropyl;

more preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is propyl or isopropyl;

preferably, the radical is a fragment

Is composed of

Preferably, the radical is a fragment

Is composed of

Preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is an isopropyl group.

In a preferred embodiment of the present invention, the compound has a structure represented by any one of the following, tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts, or prodrugs thereof:

in a fourth aspect of the present invention, there is provided a process for the preparation of a compound according to the first aspect, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, which process comprises the steps of:

i) the intermediate B-1 reacts with a compound B-3 to obtain the compound shown in the formula I

And/or the presence of a gas in the atmosphere,

ii) reaction of intermediate B-2 with compound B-4 to remove the protecting group PG of carboxyl group in B-4 ₁ To obtain intermediate B-1

Wherein, ring A, R ₁ 、R ₂ 、R ₄ Is as defined in the first aspect of the invention;

R ₃ is C ₁ -C ₆ Carboxyl or C ₁ -C ₆ An amide group.

In a preferred embodiment of the invention, the step i) is carried out under the protection of nitrogen; and/or, reacting in the presence of a catalyst; preferably, the catalyst is a rhodium catalyst and a copper catalyst; more preferably, the catalyst is rhodium acetate.

In a preferred embodiment of the invention, R ₃ Is C ₁ -C ₆ A carboxyl group; preferably, R ₃ Is butyric acid.

In a preferred embodiment of the present invention, the step i) further comprises the steps of: i-1) coupling of substituent R in Compound B-3 ₃ After being protected by a carboxyl protecting group, the intermediate B-1 reacts to obtain a compound shown in a formula I protected by the carboxyl protecting group; i-2) removing the carboxyl protecting group in the compound shown in the formula I protected by the carboxyl protecting group to obtain the compound shown in the formula I.

In a preferred embodiment of the present invention, the step i) further comprises the steps of: i-3) chiral resolution of the compound of formula I.

In a preferred embodiment of the present invention, the PG is ₁ Comprising C ₁ -C ₆ Alkyl, phenyl substituted C ₁ -C ₆ An alkyl group; preferably C ₁ -C ₆ Alkyl, more preferably methyl, ethyl, propyl, tert-butyl.

In a fifth aspect, the present invention provides a pharmaceutical composition comprising: a compound according to the first aspect of the invention, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug thereof; and a pharmaceutically acceptable carrier; or a compound according to the first aspect of the invention, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug thereof; and at least one other pharmacologically active inhibitor; preferably, the other pharmacologically active inhibitors include CTLA4 antibody, PDL1 and PD 1.

A sixth aspect of the present invention provides a use of a compound represented by formula I, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, for the prophylaxis and/or treatment of inflammatory diseases, pain, cancer, metabolic diseases, urinary system diseases; and/or a pharmaceutical composition according to the fifth aspect of the invention, comprising: use of a medicament, pharmaceutical composition or formulation for the prevention and/or treatment of inflammatory diseases, pain, cancer, metabolic diseases, urological diseases for the preparation of a medicament, pharmaceutical composition or formulation for the prevention and/or treatment of inflammatory diseases, pain, cancer, metabolic diseases, urological diseases.

In a preferred embodiment of the present invention, wherein said inflammatory disease is selected from the group consisting of arthritis, rheumatoid arthritis; the pain is selected from osteoarthritis pain, pain caused by endometriosis; the cancer is selected from solid cancers, preferably, from breast cancer, cervical cancer, colorectal cancer, endometrial cancer, glioblastoma, head and neck cancer, kidney cancer, liver cancer, lung cancer, medulloblastoma, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer and urinary tract cancer; the metabolic disease is diabetes, and the urinary system disease is overactive bladder.

In a preferred embodiment of the invention, wherein the medicament is administered in combination with radiation therapy and/or antibody therapy, wherein the antibody therapy is selected from one or a combination of CTLA4 antibody therapy, PDL1 antibody therapy and PD1 antibody therapy.

Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.

Terms and definitions

Unless otherwise indicated, the definitions of groups and terms described in the specification and claims of the present application, including definitions thereof as examples, exemplary definitions, preferred definitions, definitions described in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. The definitions of the groups and the structures of the compounds in such combinations and after the combination should fall within the scope of the present specification.

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 the claimed subject matter belongs. All patents, patent applications, and publications cited herein are incorporated by reference in their entirety unless otherwise indicated. If there are multiple definitions for terms herein, those in this section prevail.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter claimed. In this application, the use of the singular also includes the plural unless specifically stated otherwise. It must be noted that, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the term "comprising" as well as other forms, such as "includes," "including," and "containing," are not limiting.

Definitions for standardized chemical terms can be found in the literature references, including Carey and Sundberg, "ADVANCED ORGANIC chemical 4 they," vols.a (2000) and B (2001), Plenum Press, New York. Unless otherwise indicated, conventional methods within the skill of the art are employed, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy, and pharmacological methods. Unless a specific definition is set forth, the terminology used herein in the pertinent description of analytical chemistry, organic synthetic chemistry, and pharmaceutical and medicinal chemistry is known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of patients. For example, the reaction and purification can be carried out using the instructions of the kit from the manufacturer, or according to the methods known in the art or the instructions of the present invention. The techniques and methods described above can generally be practiced according to conventional methods well known in the art, as described in various general and more specific documents referred to and discussed in this specification. In the present specification, groups and substituents thereof may be selected by one skilled in the art to provide stable moieties and compounds.

When a substituent is described by a general formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the formula is written from right to left. For example, CH ₂ O is equal to OCH ₂ . As used herein, the term "a" or "an" refers to,

represents the attachment site of the group.

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including, but not limited to, patents, patent applications, articles, books, operating manuals, and treatises, are hereby incorporated by reference in their entirety.

In addition to the foregoing, when used in the specification and claims of this application, the following terms take the meanings indicated below, unless otherwise specifically indicated.

Where numerical ranges are recited in the specification and claims of this application, and where numerical ranges are understood to be "integers," they are understood to recite both the endpoints of the ranges as well as each and every integer within the ranges. For example, "an integer of 1 to 6" should be understood to describe each integer of 0, 1,2,3,4, 5, and 6.

In the present application, the term "halogen", alone or as part of another substituent, means fluorine, chlorine, bromine, iodine.

As used herein, the term "amino", alone or as part of another substituent, denotes-NH ₂ 。

As used herein, the term "nitro", alone or as part of another substituent, denotes-NO ₂ 。

As used herein, the term "cyano," alone or as part of another substituent, denotes — CN.

As used herein, the term "alkyl", alone or as part of another substituent, means a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, free of unsaturation, having, for example, 1 to 6 carbon atoms, and attached to the rest of the molecule by single bonds. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, and hexyl. An alkyl group may be unsubstituted or substituted with one or more suitable substituents. The alkyl group may also be an isotopic isomer of the naturally abundant alkyl group enriched in carbon and/or hydrogen isotopes (i.e., deuterium or tritium). As used herein, the term "alkenyl" refers to an unbranched or branched monovalent hydrocarbon chain containing one or more carbon-carbon double bonds. As used herein, the term "alkynyl" refers to an unbranched or branched, monovalent hydrocarbon chain containing one or more carbon-carbon triple bonds.

The term "C" alone or as part of another substituent ₁ -C ₆ Alkyl "is understood to mean a straight-chain or branched, saturated monovalent hydrocarbon radical having 1,2,3,4, 5 or 6 carbon atoms. The alkyl group is, for example, a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, an isopropyl group, an isobutyl group, a sec-butyl group, a tert-butyl group, an isopentyl group, a 2-methylbutyl group, a 1-ethylpropyl group, a 1, 2-dimethylpropyl group, a neopentyl group, a 1, 1-dimethylpropyl group, a 4-methylpentyl group, a 3-methylpentyl group, a 2-methylpentyl group, a 1-methylpentyl group, a 2-ethylbutyl group, a 1-ethylbutyl group, a 3, 3-dimethylbutyl group, a2, 2-dimethylbutyl group, a 1, 1-dimethylbutyl group, a2, 3-dimethylbutyl group, a 1, 3-dimethylbutyl group or a 1, 2-dimethylbutyl group, or isomers thereof. In particular, the radicals have 1,2 or 3 carbon atoms ("C) ₁ -C ₃ Alkyl groups) such as methyl, ethyl, n-propyl or isopropyl.

The term "cycloalkyl" or "carbocyclyl", alone or as part of another substituent, refers to a cyclic alkyl group. The term "m-n membered cycloalkyl" or "C _m -C _n Cycloalkyl "is understood to mean a saturated, unsaturated or partially saturated carbocyclic ring having m to n atoms. For example, "3-15 membered cycloalkyl" or "C ₃ -C ₁₅ Cycloalkyl "refers to a cyclic alkyl group containing 3 to 15, 3 to 9, 3 to 6, or 3 to 5 carbon atoms, which may contain 1 to 4 rings. "5-8 membered cycloalkyl" then contains 5-8 carbon atoms. Including monocyclic, bicyclic, tricyclic, spiro, or bridged rings. Examples of unsubstituted cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and adamantyl, or bicyclic hydrocarbon groups such as a decalin ring. The cycloalkyl group may be substituted with one or more substituents. In some embodiments, the cycloalkyl group can be a cycloalkyl group fused to an aryl or heteroaryl group. The term "C ₃ -C ₆ Cycloalkyl "is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring, whichHas 3 to 6 carbon atoms, including fused or bridged polycyclic ring systems. For example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

The term "C" alone or as part of another substituent ₂ -C ₆ Alkenyl "is understood to mean a straight-chain or branched monovalent hydrocarbon radical comprising one or more double bonds and having, for example, 2,3,4, 5 or 6 carbon atoms (i.e. C) ₂ -C ₆ Alkenyl) or having 2 or 3 carbon atoms (i.e., C) ₂ -C ₃ Alkenyl). It is understood that where the alkenyl group contains more than one double bond, the double bonds may be separated from each other or conjugated. The alkenyl group is, for example, vinyl, allyl, (E) -2-methylvinyl, (Z) -2-methylvinyl, (E) -but-2-enyl, (Z) -but-2-enyl, (E) -but-1-enyl, (Z) -but-1-enyl, pent-4-enyl, (E) -pent-3-enyl, (Z) -pent-3-enyl, (E) -pent-2-enyl, (Z) -pent-2-enyl, (E) -pent-1-enyl, (Z) -pent-1-enyl, hex-5-enyl, (E) -hex-4-enyl, (Z) -hex-4-enyl, (E) -hex-3-enyl, (Z) -hex-3-enyl, (E) -hex-2-enyl, (Z) -hex-2-enyl, (E) -hex-1-enyl, (Z) -hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E) -1-methylprop-1-enyl, (Z) -1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E) -2-methylbut-2-enyl, (Z) -2-methylbut-2-enyl, (E) -1-methylbut-2-enyl, (Z) -1-methylbut-2-enyl, (E) -3-methylbut-1-enyl, (Z) -3-methylbut-1-enyl, (E) -2-methylbut-1-enyl, (Z) -2-methylbut-1-enyl, (E) -1-methylbut-1-enyl, (Z) -1-methylbut-1-enyl, 1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, m-methylbut-2-enyl, m-methylbut-1-enyl, m-methylbut-2-enyl, m-methylbut-1-ylprop-1-enyl, m-methylbut-2-enyl, m-methylbut-1-enyl, m, 1-propylvinyl group and 1-isopropylvinyl group.

The term "C" alone or as part of another substituent ₂ -C ₆ Alkynyl "is understood to mean a straight-chain or branched monovalent hydrocarbon radical comprising one or more triple bonds and having, for example, 2,3,4, 5 or 6 carbon atoms (i.e.," C ₂ -C ₆ Alkynyl "), or having 2 or 3 carbon atoms (" C) ₂ -C ₃ Alkynyl "). Said alkynyl isFor example ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but-2-ynyl, but-3-ynyl, pent-1-ynyl, pent-2-ynyl, pent-3-ynyl, pent-4-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, hex-4-ynyl, hex-5-ynyl, 1-methylprop-2-ynyl, 2-methylbut-3-ynyl, 1-methylbut-2-ynyl, 3-methylbut-1-ynyl, 1-ethylprop-2-ynyl, 3-methylpent-4-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-3-ynyl, 1-methylpent-3-ynyl, 4-methylpent-2-ynyl, 1-methylpent-2-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-ethylbut-3-ynyl, 1-ethylbut-2-ynyl, 1-propylprop-2-ynyl, 1-isopropylprop-2-ynyl, 2-dimethylbut-3-ynyl, 2-methylpent-4-ynyl, 1-methylpent-4-ynyl, 2-methylpent-1-ynyl, 4-methylpent-1-ynyl, 3-methylpent-1-ynyl, 2-isopropylprop-2-ynyl, 2-dimethylbut-3-ynyl, 2-methylbut-3-ynyl, 2-methylpent-2-ynyl, 2-methylpent-ynyl, and the like, 1, 1-dimethylbut-3-ynyl, 1-dimethylbut-2-ynyl or 3, 3-dimethylbut-1-ynyl. In particular, the alkynyl group is ethynyl, prop-1-ynyl or prop-2-ynyl.

"haloalkyl", alone or as part of another substituent, is intended to include both branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms substituted with one or more halogens (e.g., -CvFw, where v ═ 1 to 3 and w ═ 1 to (2v + 1)). Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2,2, 2-trifluoroethyl, heptafluoropropyl, and heptachloropropyl.

The term "7-to 10-membered bridged ring" means a cyclic hydrocarbon in which any two rings share two carbon atoms which are not directly connected in the compound, and is classified into a bicyclic hydrocarbon, a tricyclic hydrocarbon, a tetracyclic hydrocarbon, etc., according to the number of constituting rings, and contains 7 to 10 carbon atoms. Non-limiting examples include:

the term "ester group", alone or as part of another substituent, refers to a group having the formula R-CO-O-or-CO-O-R, wherein R is alkyl, and alkyl is as defined herein, e.g., "C ₂ -C ₆ The "ester group" means C ₁ -C ₅ Radicals of the alkyl-CO-O-structure or-CO-O-C ₁ -C ₅ Representative examples of alkyl structures, ester groups include (but are not limited to): CH (CH) ₃ COO-、C ₂ H ₅ COO-、C ₃ H ₈ COO-、(CH ₃ ) ₂ CHCOO-、-COOCH ₃ 、-COOC ₂ H ₅ 、-COOC ₃ H ₈ Or the like.

The term "amido", alone or as part of another substituent, refers to a group having the formula-CO-NH-or-CO-NH-R, wherein R is alkyl, and alkyl is as defined herein, e.g., "C ₁ -C ₄ Amido "is C ₁ -C ₃ Radicals of alkyl-CO-NH-structure or-CO-NH-C ₁ -C ₃ Radicals of alkyl structure or-CO-NH ₂ Representative examples of ester groups include (but are not limited to): CH (CH) ₃ CO-NH-、C ₂ H ₅ -CO-NH-、C ₃ H ₈ -CO-NH-、(CH ₃ ) ₂ -CO-NH-、-CO-NH-CH ₃ 、-CO-NHC ₂ H ₅ 、-CO-NH-C ₃ H ₈ Or the like.

The term "inert solvent" includes, but is not limited to: toluene, benzene, water, methanol, ethanol, isopropanol, ethylene glycol, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran dichloromethane, chloroform, 1, 2-dichloroethane, acetonitrile, N-dimethylformamide, N-dimethylacetamide, dioxane, or a combination thereof.

The compounds provided herein, including intermediates useful in the preparation of the compounds provided herein, contain reactive functional groups (such as, but not limited to, carboxyl, hydroxyl, and amino moieties), and include protected derivatives thereof. "protected derivatives" are those compounds in which one or more reactive sites are blocked by one or more protecting groups (also referred to as protecting groups). Suitable protecting groups for the carboxyl moiety include benzyl, t-butyl, and the like, as well as isotopes and the like. Suitable amino and amido protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl, benzyloxycarbonyl and the like. Suitable hydroxyl protecting groups include benzyl and the like. Other suitable protecting groups are well known to those of ordinary skill in the art.

In this application, "optional," "optionally," "preferably," or "preferably" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optionally substituted aryl" means that the aryl group is substituted or unsubstituted, and the description includes both substituted and unsubstituted aryl groups.

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

"pharmaceutically acceptable acid addition salts" refers to salts with inorganic or organic acids which retain the biological effectiveness of the free base without other side effects. "pharmaceutically acceptable base addition salts" refers to salts with inorganic or organic bases which maintain the biological effectiveness of the free acid without other side effects. In addition to pharmaceutically acceptable salts, other salts are also contemplated by the present invention. They may serve as intermediates in the purification of the compounds or in the preparation of other pharmaceutically acceptable salts or may be used in the identification, characterization or purification of the compounds of the invention.

The term "amine salt" refers to the product resulting from the neutralization of an alkyl primary, secondary or tertiary amine with an acid. The acid includes an inorganic acid or an organic acid as described herein.

The term "stereoisomer" refers to isomers resulting from the different arrangement of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers, and conformers.

Depending on the choice of starting materials and processes, the compounds according to the invention may be present in the form of one of the possible isomers or of a mixture thereof, for example as pure optical isomers, or as isomer mixtures, for example as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule with respect to the chiral center (or centers) in the molecule. 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.

When the bond to a chiral carbon in the formula of the invention is depicted as a direct line, it is to be understood that both the (R) and (S) configurations of the chiral carbon and the resulting enantiomerically pure compounds and mixtures thereof are encompassed within the formula. The enantiomers or enantiomerically pure compounds herein are illustrated by Maehr, J.chem.Ed.1985, 62: 114-120. The absolute configuration of a stereocenter is represented by wedge bonds and dashed bonds.

The term "tautomer" refers to an isomer of a functional group resulting from the rapid movement of an atom in two positions in a molecule. The compounds of the invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually result in a mixture whose physicochemical properties are consistent with the mixture of the compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; whereas in phenol the enol type predominates. The present invention encompasses all tautomeric forms of the compounds.

In the present application, "pharmaceutical composition" refers to a formulation of a compound of the present invention with a vehicle generally accepted in the art for delivering biologically active compounds to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to facilitate administration to a living body, facilitate absorption of the active ingredient, and exert biological activity.

As used herein, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonizing agent, solvent, or emulsifier that is approved by the relevant governmental regulatory agency for human or livestock use.

The term "solvate" means that the compound of the present invention or a salt thereof includes a stoichiometric or non-stoichiometric amount of solvent bonded with non-covalent intermolecular forces, and when the solvent is water, it is a hydrate.

The term "prodrug" refers to a compound of the invention that can be converted to a biologically active compound under physiological conditions or by solvolysis. Prodrugs of the invention are prepared by modifying functional groups in the compounds, which modifications may be routinely made or removed in vivo to give the parent compound. Prodrugs include compounds of the present invention wherein a hydroxy or amino group is attached to any group that, when administered to a mammalian subject, cleaves to form a free hydroxy or a free amino group, respectively.

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may be labelled with radioisotopes, such as deuterium (g) ((R)) ² H) Tritium (A) ³ H) Iodine-125 ( ¹²⁵ I) Or C-14( ¹⁴ C) In that respect All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.

The term "adjuvant" refers to a pharmaceutically acceptable inert ingredient. Examples of classes of the term "excipient" include, without limitation, binders, disintegrants, lubricants, glidants, stabilizers, fillers, diluents, and the like. Excipients enhance the handling characteristics of the pharmaceutical formulation, i.e., make the formulation more amenable to direct compression by increasing flowability and/or adhesiveness.

As used herein, the term "treatment" and other similar synonyms include the following meanings:

(i) preventing the occurrence of a disease or condition in a mammal, particularly when such mammal is predisposed to the disease or condition but has not yet been diagnosed as having the disease or condition;

(ii) inhibiting the disease or disorder, i.e., arresting its development;

(iii) alleviating the disease or condition, i.e., causing regression of the state of the disease or condition; or

(iv) Alleviating the symptoms caused by the disease or disorder.

The reaction temperature in the reaction of each step may be suitably selected depending on the solvent, starting material, reagent, etc., and the reaction time may be suitably selected depending on the reaction temperature, solvent, starting material, reagent, etc. After the reaction in each step, the target compound may be separated and purified from the reaction system by a conventional method, such as filtration, extraction, recrystallization, washing, silica gel column chromatography, etc. Under the condition of not influencing the next reaction, the target compound can directly enter the next reaction without separation and purification.

Advantageous effects

The present inventors have made extensive and intensive studies and have unexpectedly developed a fused ring compound which is an EP4 antagonist, and which has a potent antagonistic action against EP4 and a good affinity for EP4 receptor. In addition, the invention provides a fused ring compound as an EP4 antagonist, an intermediate for preparing the compound and a preparation method, wherein the method is simple to operate, high in yield and purity and can be used for industrial production of medicines.

Detailed Description

The present invention is further illustrated by the following examples. It is to be understood that the following description is only of the most preferred embodiments of the present invention and should not be taken as limiting the scope of the invention. In the following examples, the experimental methods without specific conditions, usually according to the conventional conditions or according to the conditions suggested by the manufacturers, can be modified by those skilled in the art without essential changes, and such modifications should be considered as included in the protection scope of the present invention.

The compounds of the formula I according to the invention can be prepared by the synthesis methods described below, in which the substituents of the general formulae have the meanings given above. These methods are intended to illustrate the invention without limiting its subject matter and the scope of the compounds claimed for these examples. The invention relates to a fused ring compound as an EP4 antagonist, a preparation method and application thereof, wherein the method comprises any one of the following synthetic routes:

when ring A is absent, intermediate B-1 has the structure

It is prepared by the following route:

mixing the intermediate B-6 with trimethyl sulfonium iodide and sodium hydrogen to react to obtain an intermediate B-5, and removing carboxyl protecting group PG ₂ To obtain an intermediate B-1. The protective group PG for carboxyl group ₂ Comprising C ₁ -C ₆ Alkyl, phenyl substituted C ₁ -C ₆ An alkyl group; preferably C ₁ -C ₆ Alkyl, more preferably methyl, ethyl, propyl, tert-butyl.

In the above formulae, R ₃ Is C ₁ -C ₆ Carboxyl or C ₁ -C ₆ An amide group; preferably, R ₃ Is C ₁ -C ₆ A carboxyl group; more preferably, R ₃ Is butyric acid. The method further comprises subjecting the R to ₃ And after the protection of carboxyl protecting group, reacting with the intermediate B-1 to obtain the compound shown in the formula I protected by the carboxyl protecting group, and removing the carboxyl protecting group in the compound shown in the formula I protected by the carboxyl protecting group to obtain the compound shown in the formula I. The carboxyl protecting group comprises C ₁ -C ₆ Alkyl, phenyl substituted C ₁ -C ₆ An alkyl group; preferably C ₁ -C ₆ Alkyl, more preferably methyl, ethyl, propyl, tert-butyl.

The step i) is optionally reacted under the protection of nitrogen; and/or in the presence of a catalyst; among them, preferred catalysts are rhodium catalysts and copper catalysts, and more preferred catalyst is rhodium acetate.

According to different groups in the compound of the formula I, different synthetic routes and intermediates can be selected, when an active group (such as carboxyl, amino, hydroxyl and the like) exists in a substituent, the active group can participate in the reaction after being protected by a protecting group according to needs, and the protecting group is removed after the reaction is finished. Those compounds in which one or more reactive sites are blocked with one or more protecting groups (also referred to as protecting groups) are "protected derivatives" of the compounds of formula I as described herein. For example, suitable protecting groups for the carboxyl moiety include benzyl, t-butyl, methyl, ethyl, and the like. Suitable amino and amido protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable hydroxyl protecting groups include benzyl and the like. Other suitable protecting groups are well known to those of ordinary skill in the art.

Preferably, the reaction requires inert gas shielding, including but not limited to: nitrogen, helium, neon, argon.

The reactions of the various steps of the present invention are preferably carried out in an inert solvent including, but not limited to: toluene, benzene, water, methanol, ethanol, isopropanol, ethylene glycol, N-methylpyrrolidone, dimethyl sulfoxide, tetrahydrofuran dichloromethane, chloroform, 1, 2-dichloroethane, acetonitrile, N-dimethylformamide, N-dimethylacetamide, dioxane, or a combination thereof.

Abbreviations are defined as follows:

symbol or unit:

IC ₅₀ : the median inhibitory concentration is the concentration at which half of the maximum inhibitory effect is achieved

M: for example, n-butyllithium (14.56mL, 29.1mmol, 2.5M solution in n-hexane) represents a 2.5mol/L n-hexane solution of n-butyllithium

N: equivalent concentration, e.g. 2N hydrochloric acid means 2mol/L hydrochloric acid solution

Reagent:

DCM: methylene dichloride

DIPEA: also can be written as DIEA, diisopropylethylamine, i.e., N-diisopropylethylamine

DMF: n, N-dimethylformamide

DMSO, DMSO: dimethyl sulfoxide

EA: acetic acid ethyl ester

Et ₃ N: triethylamine

MeOH: methanol

PE: petroleum ether

THF: tetrahydrofuran (THF)

Test or detection methods:

HPLC: high performance liquid chromatography

SFC: supercritical fluid chromatography

Example 1: synthesis of Compound I-1

The synthetic route is as follows:

the first step is as follows: synthesis of 3-bromo-4- (hydroxymethyl) benzoic acid (B1-2)

The compound methyl 3-bromo-4- (hydroxymethyl) benzoate (B1-1) (1.0g,4.08mmol) was added to tetrahydrofuran (20mL) and water (20mL) at room temperature, sodium hydroxide (0.5g,12.5mmol) was added, and the mixture was stirred for 12 h. pH was adjusted to 3 with 1N hydrochloric acid, extracted with ethyl acetate (20mL × 3), separated, the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give the compound 3-bromo-4- (hydroxymethyl) benzoic acid (crude white solid, 1.0g, 100% yield).

The second step is that: synthesis of 3-bromo-4- (hydroxymethyl) -N-isopropylbenzamide (B1-3)

The compound 3-bromo-4- (hydroxymethyl) benzoic acid (1.0g,4.08mmol) was added to dichloromethane (30mL) at room temperature, isopropylamine (300mg,5.08mmol), 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (1.6g,4.9mmol), N, N-diisopropylethylamine (1.05g,8.1mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (50mL) was added for dilution, and extraction was performed with dichloromethane (30mL × 3), liquid separation was performed, organic phases were combined, the organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 4:1) to obtain compound 3-bromo-4- (hydroxymethyl) -N-isopropylbenzamide (crude colorless liquid, 0.8g, yield 72%). LC-MS, M/Z (ESI):272.2[ M + H] ⁺ 。

The third step: synthesis of 4- ((allyloxy) methyl) -3-bromo-4- (hydroxymethyl) -N-isopropylbenzamide (B1-4)

The compound 3-bromo-4- (hydroxymethyl) -N-isopropylbenzamide (800mg,2.94mmol) was added to tetrahydrofuran (20mL) at room temperature, 60% sodium hydride (488mg,12.2mmol) was added, stirring was carried out at room temperature for 0.5h, allyl bromide (532mg,4.4mmol) was added, and stirring was carried out at room temperature for 3 h. Water (30mL) was added for dilution, extraction with ethyl acetate (20mL × 3), liquid separation, organic phases were combined, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 4- ((allyloxy) methyl) -3-bromo-4- (hydroxymethyl) -N-isopropylbenzamide (crude white solid, 1.0g, 100% yield). LC-MS, M/Z (ESI):312.2[ M + H] ⁺ 。

The fourth step: synthesis of N-isopropyl-4-methylidene-6-carboxamide (B1-5)

The compound 4- ((allyloxy) methyl) -3-bromo-4- (hydroxymethyl) -N-isopropylbenzamide (1.0g,3.2mmol) was added to N, N-dimethylacetamide (10mL) at room temperature, cesium carbonate (2.0g,6.4mmol), palladium tetratriphenylphosphine (7.39mg,0.64mmol), tetrabutylammonium iodide (1.18g,3.2mmol) were added, heated to 100 ℃ under nitrogen and stirred for 16 h. Cooling to room temperature, diluting with water (50mL), extracting with ethyl acetate (30mL × 3), separating, combining the organic phases, drying the organic phases over anhydrous sodium sulfate, filtering, concentrating, and separating and purifying the residue with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 10:1) to obtain the compound N-isopropyl-4-methyleneisochroman-6-carboxamide (crude white solid, 350mg, 47% yield). LC-MS, M/Z (ESI) 232.3[ M + H ]] ⁺ 。

The fifth step: synthesis of ethyl 6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochromane ] -2-carboxylate (B1-6)

The starting material, N-isopropyl-4-methylidene isochromane-6-carboxamide (300mg,1.3mmol), was added to dichloromethane (20mL) at room temperature, rhodium acetate (265mg,0.6mmol) was added under nitrogen, heated to 50 deg.C, ethyl diazoacetate (0.5mL,6.0mmol) was added, and stirred for 0.5 h. Cooling to room temperature, concentrating, separating and purifying the residue with silica gel column (petroleum ether: ethyl acetate (V/V) ═ 5:1) to obtain compound 6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochromane]-ethyl 2-carboxylate (crude white solid, 80mg, 20% yield). LC-MS, M/Z (ESI) 318.2[ M + H] ⁺ 。

And a sixth step: synthesis of 6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochromane ] -2-carboxylic acid (B1-7)

Compound 6' - (isopropylmethyle) is reacted at room temperatureAmide) spiro [ cyclopropane-1, 4' -isochromane]Ethyl-2-carboxylate (38mg,0.12mmol) was added to tetrahydrofuran (5mL) and water (5mL), lithium hydroxide (15mg,0.36mmol) was added, and the mixture was stirred at room temperature for 16 h. Adjusting pH to 3 with 1N hydrochloric acid, diluting with water (10mL), extracting with ethyl acetate (10mL × 3), separating, combining organic phases, drying the organic phase with anhydrous sodium sulfate, filtering, and concentrating to obtain compound 6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochromane]-2-carboxylic acid (crude white solid, 40mg, 100% yield). LC-MS, M/Z (ESI):290.3[ M + H] ⁺ 。

The seventh step: synthesis of ethyl 4- (4-cyano-2- ((1S,2R) -6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochroman ] -2-carboxamide) phenyl) butyrate (B1-8)

At room temperature, the compound 6'- (isopropyl formamide) spiro [ cyclopropane-1, 4' -isochromane]-2-Carboxylic acid (28mg,0.096mmol) was added to 5mL dichloromethane, pyridine (0.25mL) was added, ethyl 4- (2-amino-4-cyanophenyl) butyrate (23mg,0.10mmol) was added, cooled to 0 deg.C, 1 drop of phosphorus oxychloride was added, and stirred at room temperature for 16 h. Diluting with water (30mL), extracting with ethyl acetate (50 mL. times.3), separating, combining organic phases, drying the organic phases with anhydrous sodium sulfate, filtering, and concentrating to obtain compound 4- (4-cyano-2- ((1S,2R) -6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochromane]-2-carboxamide) phenyl) butyric acid ethyl ester (white solid, 22mg, yield 45%). LC-MS, M/Z (ESI):504.3[ M + H] ⁺ 。

Eighth step: synthesis of 4- (4-cyano-2- ((1S,2R) -6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochroman ] -2-carboxamide) phenyl) butyric acid (I-1)

Ethyl 4- (4-cyano-2- ((1S,2R) -6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochroman ] -2-carboxamide) phenyl) butanoate (22mg,0.43mmol) was added to tetrahydrofuran (5mL) and water (5mL) at room temperature, lithium hydroxide (8.9mg,0.21mmol) was added, and the mixture was stirred at room temperature for 4 h. The compound 4- (4-cyano-2- ((1S,2R) -6'- (isopropylformamide) spiro [ cyclopropane-1, 4' -isochroman ] -2-carboxamide) phenyl) butyric acid (white solid, 4.5mg, 22% yield) was prepared by adjusting pH to 3 with 1N hydrochloric acid, concentrating, subjecting the residue to an acidic preparation method a (acidic preparation method a: Welch, Ultimate C18 column, 10 μm, 21.2mm × 250mm, pure water solution of trifluoroacetic acid with mobile phase a of 1 ‰, acetonitrile solution of mobile phase B, gradient conditions of 0 to 3 minutes, mobile phase a remaining 90%, gradient elution of 3 to 18 minutes, 90% to 5%, and 5% remaining 18 to 22 minutes).

¹ H NMR(400mHz,DMSO-d6)δ10.33(s,1H),8.40(t,1H),8.96(s,1H),7.67(q,1H),7.56(q,1H),7.45(t,2H),7.15(d,1H),4.83(d,2H),4.12-4.04(m,2H),3.92(d,1H),2.70-2.64(m,3H),2.54(s,1H),2.16(t,2H),1.71(t,3H),1.56(t,1H),1.19(d,6H)。

LC-MS,M/Z(ESI):476.3[M+H] ⁺ 。

Example 2: synthesis of Compound I-2

The synthetic route is as follows:

the first step is as follows: synthesis of 6-bromo-4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene (B2-2)

The compound 6-bromo-2, 3-dihydro-1, 3-methano-4 (1H) -one (1.48g,6.25mmol) was added to tetrahydrofuran (30mL) at room temperature and μ -chloro- μ -methylenebis [ cyclopentadienyl ] titanium ] -dimethylaluminum (3.56g,12.5mmol) was added and stirred for 12H. Water (50mL) was added to dilute the mixture, and the mixture was extracted with ethyl acetate (30mL × 3), separated, the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 1:1) to give the compound 6-bromo-4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene (B2-2) (crude white solid, 850mg, yield 58%).

The second step: synthesis of ethyl 4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxylate (B2-3)

The compound 6-bromo-4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene (910mg,3.89mmol) was added to ethanol (50mL) at room temperature, triethylamine (1.18g,11.7mmol) was added, 1' -bis-diphenylphosphino ferrocene palladium dichloride (477mg,0.58mmol) was added, carbon monoxide was added, heated to 80 ℃ and stirred for 16 h. Cooling to room temperature, diluting with water (100mL), extracting with ethyl acetate (50mL × 3), separating, combining the organic phases, drying the organic phases over anhydrous sodium sulfate, filtering, concentrating, and separating and purifying the residue with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 1:1) to give the compound ethyl 4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxylate (B2-3) (crude white solid, 870mg, yield 98%). LC-MS, M/Z (ESI) 229.2[ M + H ]] ⁺ 。

The third step: synthesis of 4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxylic acid (B2-4)

The compound ethyl 4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxylate (705mg,3.1mmol) was added to tetrahydrofuran (8mL) and water (8mL) at room temperature, lithium hydroxide (650mg,15.5mmol) was added, and the mixture was stirred at room temperature for 16 h. pH was adjusted to 3 with 1N hydrochloric acid, water (30mL) was added for dilution, extraction was performed with ethyl acetate (20mL × 3), liquid separation was performed, organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtration and concentration were performed, and the residue was separated and purified with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 1:1) to obtain compound 4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxylic acid (B2-4) (crude white solid, 620mg, yield 100%). LC-MS, M/Z (ESI) 201.2[ M + H ]] ⁺ 。

The fourth step: synthesis of N-isopropyl-4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxamide (B2-5)

The compound 4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxylic acid (620mg,3.1mmol) was added to DMF (20mL) at room temperature, isopropylamine (201mg,3.4mmol), 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (1.53g,4.0mmol), N, N-diisopropylethylamine (1.2g,9.3mmol) was added, and stirred at room temperature for 16 h. Water (50mL) was added for dilution, and extraction was performed with EA (30mL × 3), followed by liquid separation, the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 1:1) to obtain a compound, N-isopropyl-4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxamide (B2-5) (crude white solid, 750mg, yield 100%). LC-MS, M/Z (ESI) 242.2[ M + H ]] ⁺ 。

The fifth step: synthesis of ethyl 6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxylate (B2-6)

The starting material N-isopropyl-4-methylene-1, 2,3, 4-tetrahydro-1, 3-methylnaphthalene-6-carboxamide (700mg,2.9mmol) was added to dichloromethane (10mL) at room temperature, ethyl diazoacetate (1.65g,14.5mmol), rhodium acetate (257mg,0.58mmol) were added, heated to 40 ℃ and stirred for 16 h. Cooling to room temperature, diluting with water (50mL), extracting with DCM (30mL × 3), separating, combining the organic phases, drying the organic phase over anhydrous sodium sulfate, filtering, concentrating, and separating and purifying the residue with silica gel column (petroleum ether: ethyl acetate (V/V) ═ 1:1) to give 6'- (isopropylformamide) -2',3 '-dihydro-1' H-spiro [ cyclopropane-1, 4'- [1, 3' ] -1, 3]]Methylnaphthalene]-ethyl 2-carboxylate (B2-6) (crude white solid, 560mg, 59% yield). LC-MS, M/Z (ESI):328.2[ M + H] ⁺ 。

And a sixth step: synthesis of 6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxylic acid (B2-7)

The compound 6'- (isopropyl formamide) -2',3 '-dihydro-1' H-spiro [ cyclopropane-1, 4'- [1, 3' ] at room temperature]Methylnaphthalene]Ethyl-2-carboxylate (560mg,1.71mmol) was added to tetrahydrofuran (6mL) and water (6mL), lithium hydroxide (360mg,8.56mmol) was added, and the mixture was stirred at room temperature for 16 h. Adjusting the pH to 3 with 1N hydrochloric acid, diluting with water (30mL), extracting with ethyl acetate (20mL × 3), separating the liquids, combining the organic phases, drying the organic phases with anhydrous sodium sulfate, filtering, concentrating, and separating and purifying the residue with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 1:1) to obtain the compound 6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3]]Methylnaphthalene]-2-carboxylic acid (B2-7) (crude white solid, 405mg, 79% yield). LC-MS, M/Z (ESI):300.2[ M + H] ⁺ 。

The seventh step: synthesis of ethyl 4- (4-cyano-2- ((1S,2R) -6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxamide) phenyl) butyrate (B2-8)

The compound 6'- (isopropyl formamide) -2',3 '-dihydro-1' H-spiro [ cyclopropane-1, 4'- [1, 3' ] at room temperature]Methylnaphthalene]-2-Carboxylic acid (322mg,1.1mmol) was added to pyridine (10mL), ethyl 4- (2-amino-4-cyanophenyl) butanoate (255mg,1.1mmol) was added, cooled to 0 deg.C, phosphorus oxychloride (843mg,5.5mmol) was added, and stirred at room temperature for 16 h. Diluting with water (30mL), extracting with ethyl acetate (10mL × 3), separating, combining organic phases, drying the organic phases with anhydrous sodium sulfate, filtering, concentrating, and separating and purifying the residue with silica gel column (petroleum ether: ethyl acetate (V/V) ═ 2:1) to obtain 4- (4-cyano-2- ((1S,2R) -6'- (isopropylformamide) -2',3 '-dihydro-1' H-spiro [ cyclopropane-1, 4'- [1, 3' ] -4]Methylnaphthalene]-2-carboxamide) phenyl) butyric acid ethyl ester (B2-8) (white solid, 300mg, yield 54%). LC-MS, M/Z (ESI):514.3[ M + H] ⁺ 。

Eighth step: synthesis of 4- (4-cyano-2- ((1S,2R) -6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxamide) phenyl) butyric acid (I-2)

The compound ethyl 4- (4-cyano-2- ((1S,2R) -6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxamide) phenyl) butanoate (300mg,0.58mmol) was added to tetrahydrofuran (6mL) and water (6mL) at room temperature, lithium hydroxide (50mg,1.2mmol) was added, and the mixture was stirred at room temperature for 16H. Water (30mL) was added to dilute the solution, and the solution was extracted with ethyl acetate (20mL × 3), separated, the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column separation (petroleum ether: ethyl acetate (V/V) ═ 2:1) to give compound 4- (4-cyano-2- ((1S,2R) -6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxamide) phenyl) butyric acid (I-2) (white solid, 120mg, yield 42%).

¹ H NMR(400mHz,DMSO-d6)δ12.09(s,1H),9.59(s,1H),7.73(t,3H),7.52-7.48(m,2H),7.32(d,1H),7.08(d,1H),4.08-3.99(m,1H),3.27(t,1H),2.46(t,4H),2.24(t,1H),2.07(t,1H),1.99-1.86(m,4H),1.48-1.39(m,4H),1.12-1.06(m,6H)。LC-MS,M/Z(ESI):486.3[M+H] ⁺ 。

Example 3: synthesis of Compound I-3

The synthetic route is as follows:

the first step is as follows: (E) synthesis of methyl (E) -3- (3- (tert-butoxy) -3-oxopropan-1-en-1-yl) benzoate (B3-2)

Compound 3-bromo-benzoic acid methyl ester (2.5g,23.4mmol) was added to toluene (30mL) at room temperature, tert-butyl acrylate (4.5g,35.1mmol), palladium bis (triphenylphosphine) dichloride (0.4g,0.58mmol), triethylamine (4.8mL,35mmol) were added, heated to 100 ℃ under nitrogen and stirred for 16 h. Cooled to room temperature, water (30mL) was added, extraction was performed with ethyl acetate (30 mL. times.3), liquid separation was performed, the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified (pure petroleum ether) with a silica gel column to give the compound (E) -methyl 3- (3- (tert-butoxy) -3-oxopropan-1-en-1-yl) benzoate (B3-2) (crude white solid, 1.3g, 43% yield).

The second step: synthesis of methyl 3- (2- (tert-butoxycarbonyl) cyclopropyl) benzoate (B3-3)

Compound (E) -methyl 3- (3- (tert-butoxy) -3-oxopropan-1-en-1-yl) benzoate (2.3g,8.8mmol) was added to dimethylsulfoxide (50mL) and tetrahydrofuran (50mL) at room temperature, trimethylsulfoxonium iodide (3.9g,17.6mmol), 60% sodium hydride (352mg,8.8mmol) were added, and the mixture was stirred at room temperature for 16 h. Water (100mL) was added for dilution, extraction was performed with ethyl acetate (200mL × 3), liquid separation was performed, organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 20:1) to obtain compound 3- (-2- (tert-butoxycarbonyl) cyclopropyl) benzoic acid methyl ester (B3-3) (crude colorless liquid, 643mg, yield 27%). LC-MS, M/Z (ESI):277.2[ M + H] ⁺ 。

The third step: synthesis of 3- (2- (tert-butoxycarbonyl) cyclopropyl) benzoic acid (B3-4)

The compound methyl 3- (2- (tert-butoxycarbonyl) cyclopropyl) benzoate (640mg,2.3mmol) was added to tetrahydrofuran (10mL) and water (3mL) at room temperature, lithium hydroxide (290mg,6.9mmol) was added, and the mixture was stirred at room temperature for 3 hours. Adjusting pH to 7 with 1N hydrochloric acid, diluting with water (10mL), extracting with ethyl acetate (20 mL. times.3), separating, and mixingAnd the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude compound as a white solid to give the compound 3- (2- (tert-butoxycarbonyl) cyclopropyl) benzoic acid (B3-4) (crude white solid, 565mg, 93% yield). LC-MS, M/Z (ESI) 263.2[ M + H ]] ⁺ 。

The fourth step: synthesis of tert-butyl 2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxylate (B3-5)

The compound 3- (2- (tert-butoxycarbonyl) cyclopropyl) benzoic acid (565mg,2.16mmol) was added to dichloromethane (10mL) at room temperature, N, N-diisopropylethylamine (413mg,3.2mmol), 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethyluronium hexafluorophosphate (1.2g,3.2mmol), isopropylamine (153mg,2.6mmol) were added and stirred at room temperature for 2 h. Water (10mL) was added for dilution, and extraction was performed with dichloromethane (10mL × 3), liquid separation was performed, organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 3:1) to obtain compound tert-butyl 2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxylate (B3-5) (white solid, 535mg, yield 82%). LC-MS, M/Z (ESI):304.3[ M + H] ⁺ 。

The fifth step: synthesis of 2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxylic acid (B3-6)

The starting material, tert-butyl 2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxylate (535mg,1.7mmol) was added to dichloromethane (20mL) at room temperature, trifluoroacetic acid (4mL) was added and the mixture was stirred for 2 h. Concentration and the residue were separated and purified by silica gel column separation (dichloromethane: ethyl acetate (V/V) ═ 10:1) to give compound 2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxylic acid (B3-6) (crude white solid, 400mg, yield 92%). LC-MS, M/Z (ESI):248.2[ M + H] ⁺ 。

And a sixth step: synthesis of ethyl 4- (4-cyano-2- ((1R,2R) -2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxamide) phenyl) butyrate (B3-7)

Pyridine (3mL) was added to the compound 2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxylic acid (100mg,0.4mmol) at room temperature, ethyl 4- (2-amino-4-cyanophenyl) butyrate (93mg,0.4mmol) was added, cooling was performed to 0 ℃, phosphorus oxychloride (367mg,2.40mmol) was added, and stirring was performed at low temperature for 2 h. Water (30mL) was added to dilute the mixture, and the mixture was extracted with ethyl acetate (50mL × 3), separated, the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (petroleum ether: ethyl acetate (V/V) ═ 3:2) to give compound 4- (4-cyano-2- ((1R,2R) -2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxamide) phenyl) butyric acid ethyl ester (B3-7) (white solid, 130mg, yield 70%). LC-MS, M/Z (ESI):462.6[ M + H [ ]] ⁺ 。

The seventh step: synthesis of 4- (4-cyano-2- ((1R,2R) -2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxamide) phenyl) butyric acid (I-3)

The compound ethyl 4- (4-cyano-2- ((1R,2R) -2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxamide) phenyl) butyrate (130mg,0.28mmol) was added to tetrahydrofuran (5mL) and water (3mL) at room temperature, lithium hydroxide (35mg,0.84mmol) was added, and stirring was carried out at room temperature for 16 h. pH was adjusted to 7 with 1N hydrochloric acid, and the mixture was extracted with ethyl acetate (10mL × 3), separated, the organic phases were combined, the organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated and purified with a silica gel column (dichloromethane: methanol (V/V) ═ 20:1) to give compound 4- (4-cyano-2- ((1R,2R) -2- (3- (isopropylformamide) phenyl) cyclopropane-1-carboxamide) phenyl) butanoic acid (I-3) (white solid, 6.7mg, yield 5.4%).

¹ H NMR(400mHz,DMSO-d6)δ12.14(s,1H),9.83(s,1H),8.21(d,1H),8.04(s,1H),7.68(d,1H),7.67(s,1H),7.57(t,1H),7.43(dd,2H),4.14-4.06(m,2H),2.78-2.62(m,2H),2.49-2.38(m,1H),2.34(s,1H),2.24-2.20(m,2H),1.76(t,2H),1.52-1.46(m,2H),1.18(d,6H)。

LC-MS,M/Z(ESI):434.6[M+H] ⁺ 。

Example 4: synthesis of Compound I-4

The synthetic route is as follows:

the first step is as follows: synthesis of ethyl 4- (4-cyano-2- ((1S,2S) -6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxamide) phenyl) butyrate (B4-1)

The compound 6'- (isopropyl formamide) -2',3 '-dihydro-1' H-spiro [ cyclopropane-1, 4'- [1, 3' ] at room temperature]Methylnaphthalene]-2-Carboxylic acid (B2-7) (322mg,1.1mmol) was added to pyridine (10mL), ethyl 4- (2-amino-4-cyanophenyl) butanoate (255mg,1.1mmol) was added, cooled to 0 deg.C, phosphorus oxychloride (843mg,5.5mmol) was added, and stirred at room temperature for 16 h. Diluting with water (30mL), extracting with ethyl acetate (10 mL. times.3), separating, combining the organic phases, drying the organic phases with anhydrous sodium sulfate, filtering, concentrating, and separating and purifying the residue with silica gel column (petroleum ether: ethyl acetate (V/V) ═ 2:1) to obtain 4- (4-cyano-2- ((1S,2S) -6'- (isopropylformamide) -2',3 '-dihydro-1' H-spiro [ cyclopropane-1, 4'- [1, 3' ] -compound]Methylnaphthalene]-2-carboxamide) phenyl) butyric acid ethyl ester (4-1) (white solid, 240mg, yield 43%). LC-MS, M/Z (ESI) 514.3[ M + H] ⁺ 。

The second step: synthesis of 4- (4-cyano-2- ((1S,2S) -6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxamide) phenyl) butyric acid (I-4)

The compound ethyl 4- (4-cyano-2- ((1S,2S) -6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxamide) phenyl) butyrate (200mg,0.39mmol) was added to tetrahydrofuran (4mL) and water (4mL) at room temperature, lithium hydroxide (33mg,0.78mmol) was added, and the mixture was stirred at room temperature for 16H. Water (20mL) was added for dilution, extraction with ethyl acetate (10mL × 3), liquid separation, organic phases combined, organic phases dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column separation (petroleum ether: ethyl acetate (V/V) ═ 2:1) to give compound 4- (4-cyano-2- ((1S,2S) -6' - (isopropylformamide) -2',3' -dihydro-1 ' H-spiro [ cyclopropane-1, 4' - [1,3] methylnaphthalene ] -2-carboxamide) phenyl) butyric acid (I-4) (white solid, 150mg, yield 79%).

¹ H NMR(400mHz,DMSO-d6)δ12.07(s,1H),9.87(s,1H),8.13(d,1H),7.77(d,1H),7.60-7.55(m,2H),7.42(d,2H),7.13(d,1H),4.14-4.09(m,1H),3.29-3.24(m,2H),2.63-2.57(m,4H),2.42(t,1H),2.18(t,2H),1.72(t,2H),1.68-1.58(m,2H),1.52(t,2H),1.18-1.10(m,6H)。

LC-MS,M/Z(ESI):486.3[M+H] ⁺ 。

Test example 1: EP4 antagonism assay

The compounds were assayed for antagonism against EP4 in CHO stable cell lines highly expressing the human EP4 receptor. After trypsinization, the cells were resuspended in buffer (1 × HBSS, 0.1% BSA,20mM HEPES and 500 μ M IBMX) and 8000 cells per well were plated in 384-well plates in a volume of 15 μ L. Compound working solutions at 8X concentration were prepared with assay buffer, followed by addition of 2.5 μ L of 8X compound working solution to the 384 well plate described above and incubation at 37 ℃ for 30 min. Preparation of agonist PGE at 8 Xconcentration with assay buffer ₂ Working solution (4nM) was added at 2.5. mu.L per well to the 384-well plate described above (PGE) ₂ Final concentration 0.5nM), incubated at 37 ℃ for 30 min. After completion of the reaction, the amount of cAMP in the cells was quantified according to the method described in the instruction of cAMP test kit (Perkin Elmer, Cat # TRF 0263). Calculating antagonism (IC) of test Compounds ₅₀ A value).

Table 1 antagonistic action of the test compounds on EP4

Test compounds	IC 50 (nM)
		I-1	42
I-2	38
		I-3	27
I-4	>100

Experimental results show that the compound has good antagonistic action on EP 4.

Test example 2: radioligand EP4 receptor binding assay

Radioligand EP4 binding assays were performed using recombinant human EP4 receptor membrane protein (prepared from 293 cells overexpressing the human EP4 receptor). Combining the test compound with PGE ₂ Stock solutions of 10mM were prepared in DMSO solvent, and 8 concentration points of 4 Xworking solution were then diluted in a gradient of buffer (50mM HBSS, 0.1% BSA,500mM NaCl). Mixing 1 μ L of compound working solution, DMSO, PGE ₂ The working solution was added to the assay plate separately, and 100. mu.L of the EP4 receptor membrane protein (20. mu.g/well) and 100. mu.L of the radioligand [ alpha ], [ 2 ] ³ H]-PGE ₂ (Perkinelmer, Cat: NET428250UC, Lot:2469552) (final concentration 1.5nM), incubated for 1 hour at room temperature in a sealed chamber. Unifilter-96GF/C filter plates (Perkin Elmer) were soaked with 0.5% BSA, 50. mu.L/well for at least 30min at room temperature. After binding was complete, Perkin Elmer Filtermate Harv was usedester through GF/C plate filter reaction mixture, then washed filter plate, at 50 degrees C filter plate drying for 1 hours. After drying, the bottom of the filter well was sealed using Perkin Elmer Unifilter-96 sealing tape and 50. mu.L of MicroScint was added ^TM 20 cocktail (Perkin Elmer), sealing the top of the filter plate. Reading captured on filter using Perkin Elmer MicroBeta2 Reader ³ And H, counting.

Data were analyzed using GraphPad Prism 5 and inhibition was calculated according to the following formula:

inhibition (%) was 100- (test group-PGE) ₂ group)/(DMSO group-PGE ₂ Group) 100

Ki values for compound binding to EP4 as determined by radioligand EP4 binding were calculated based on inhibition at various concentrations of compound.

TABLE 2 Compound Ki values determined by radioligand EP4 binding

Test compounds	Ki(nM)
		I-1	22
I-2	94
		I-3	77
I-4	＞100

The experimental results show that the compounds of the present invention have good affinity with the EP4 receptor.

Although embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are exemplary and not to be construed as limiting the present invention, and that changes, 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 (25)

1. A compound of formula I, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug thereof:

wherein ring A is absent or 7-10 membered bridged cycloalkyl, 6 membered oxygenated heterocycloalkyl;

and when ring A is a 6-membered oxoheterocycloalkyl group,

is composed of

R ₁ Is selected from C ₁ -C ₆ Alkyl, -CONH-R ₁₁ 、-NHCO-R ₁₂ ；R ₁₁ 、R ₁₂ Each independently is hydrogen or C ₁ -C ₆ An alkyl group; r ₁ 、R ₁₁ 、R ₁₂ C as described in (1) ₁ -C ₆ Alkyl is optionally substituted by one or more R _f Substitution; when the substituent is plural, the R _f The same or different;

R ₂ selected from halogen, cyano, C ₂ -C ₆ Alkenyl radical, C ₂ -C ₆ Alkynyl, -CO-R ₂₁ ；R ₂₁ Is amino, hydroxy, C ₁ -C ₆ An alkyl group; wherein said amino or hydroxy group is optionally independently substituted by C ₁ -C ₆ Alkyl substitution; said C is ₂ -C ₆ Alkenyl radical, said C ₂ -C ₆ Alkynyl and said C ₁ -C ₆ Alkyl is optionally each independently substituted with one or more R _f Substitution; when the substituent is plural, the R _f The same or different;

R ₃ is C ₁ -C ₆ Carboxyl or C ₁ -C ₆ An amide group;

R ₄ selected from hydrogen, hydroxy, halogen, cyano, C ₁ -C ₄ Alkyl or C ₁ -C ₄ A haloalkyl group;

r mentioned above _f Selected from hydroxy, halogen or cyano.

2. The compound of claim 1, having a structure according to formula II, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug thereof:

3. the compound of claim 1 or 2, wherein ring A is absent or a 7-membered bridged cycloalkyl, a 6-membered oxoheterocycloalkyl, and when ring A is a 6-membered oxoheterocycloalkyl,

is composed of

Preferably, the first and second liquid crystal films are made of a polymer,

is composed of

When ring A is absent, a radical fragment

Is composed of

Preferably, ring A is absent

Is composed of

Preferably, when ring A is a 7-membered bridged cycloalkyl group,

is composed of

More preferably, when ring A is a 7-membered bridged cycloalkyl group,

is composed of

4. A compound according to claim 1 or 2, wherein R is ₁ is-CONH-R ₁₁ or-NHCO-R ₁₂ (ii) a Wherein R is ₁₁ 、R ₁₂ Each independently is C ₁ -C ₆ An alkyl group;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is C ₁ -C ₆ An alkyl group;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Methyl, ethyl, propyl, isopropyl;

more preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is propyl or isopropyl;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is isopropyl.

5. A compound according to claim 1 or 2, wherein R is ₂ Is halogen, cyano; preferably, R ₂ Is a cyano group.

6. A compound according to claim 1 or 2, wherein R is ₃ Is C ₁ -C ₆ A carboxyl group; preferably, R ₃ Is butyric acid.

7. A compound according to claim 1 or 2, wherein R is ₄ Selected from hydrogen, hydroxy, halogen, cyano, methyl or halomethyl; preferably, R ₄ Is hydrogen.

8. The compound of claim 1 or 2, having a structure, tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof, according to any one of the following:

9. a compound represented by formula B-1, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug thereof:

wherein ring A is absent or 7-10 membered bridged cycloalkyl, 6 membered oxygenated heterocycloalkyl;

R ₁ is selected from C ₁ -C ₆ Alkyl radical、-CONH-R ₁₁ 、-NHCO-R ₁₂ ；R ₁₁ 、R ₁₂ Each independently is hydrogen or C ₁ -C ₆ An alkyl group; r is ₁ 、R ₁₁ 、R ₁₂ C as described in (1) ₁ -C ₆ Alkyl is optionally substituted by one or more R _f Substitution; when the substituent is plural, the R _f The same or different;

R ₄ selected from hydrogen, hydroxy, halogen, cyano, C ₁ -C ₄ Alkyl or C ₁ -C ₄ A haloalkyl group;

r mentioned above _f Selected from hydroxy, halogen or cyano.

10. The compound of claim 9, having a structure according to formula B-1-1, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug thereof:

11. the compound of claim 9 or 10, wherein when ring A is a 6-membered oxoheterocycloalkyl group,

is composed of

12. The compound of claim 9 or 10,

is composed of

R ₁ is-CONH-R ₁₁ or-NHCO-R ₁₂ Wherein R is ₁₁ 、R ₁₂ Each independently is C ₁ -C ₆ Alkyl radical, R ₄ Is hydrogen;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is C ₁ -C ₆ Alkyl radical, R ₄ Is hydrogen;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is methyl, ethyl, propyl, R ₄ Is hydrogen;

more preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is propyl, R ₄ Is hydrogen;

preferably, the first and second liquid crystal films are made of a polymer,

is composed of

Preferably, the first and second air flow paths are arranged in parallel,

is composed of

Preferably, the propyl group is isopropyl; preferably, the first and second air flow paths are arranged in parallel,

is composed of

13. The compound of claim 9 or 10, having a structure, tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof, according to any of the following:

14. a compound represented by formula B-2, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt, or a prodrug thereof:

wherein, the ring A is 7-10 membered bridged cycloalkyl and 6 membered oxygen-containing heterocycloalkyl;

R ₁ is selected from C ₁ -C ₆ Alkyl, -CONH-R ₁₁ 、-NHCO-R ₁₂ ；R ₁₁ 、R ₁₂ Each independently is hydrogen or C ₁ -C ₆ An alkyl group; r ₁ 、R ₁₁ 、R ₁₂ C as described in (1) ₁ -C ₆ Alkyl is optionally substituted by one or more R _f Substitution; when the substituent is plural, the R _f The same or different;

R ₄ selected from hydrogen, hydroxy, halogen, cyano, C ₁ -C ₄ Alkyl or C ₁ -C ₄ A haloalkyl group;

r mentioned above _f Selected from hydroxyl, halogen and cyano.

15. The compound of claim 14, wherein ring A is a 6-membered oxoheterocycloalkyl,

is composed of

16. The compound of claim 14, wherein R is ₁ For the purpose ofCONH-R ₁₁ or-NHCO-R ₁₂ (ii) a Wherein R is ₁₁ 、R ₁₂ Each independently is C ₁ -C ₆ Alkyl radical, R ₄ Is hydrogen;

preferably, the first and second air flow paths are arranged in parallel,

is composed of

R ₁ is-CONH-R ₁₁ ，R ₁₁ Is C ₁ -C ₆ Alkyl radical, R ₄ Is hydrogen;

preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is methyl, ethyl, propyl, isopropyl;

more preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is propyl or isopropyl;

preferably, the first and second liquid crystal films are made of a polymer,

is composed of

Preferably, the first and second liquid crystal films are made of a polymer,

is composed of

Preferably, R ₁ is-CONH-R ₁₁ ，R ₁₁ Is isopropyl.

17. The compound of claim 14, having a structure, tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof, of any one of the following:

18. a process for preparing a compound, tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof according to any one of claims 1 to 8, characterized in that said process comprises the steps of:

i) the intermediate B-1 reacts with a compound B-3 to obtain the compound shown in the formula I

And/or the presence of a gas in the atmosphere,

ii) reaction of intermediate B-2 with compound B-4 to remove the protecting group PG of carboxyl group in B-4 ₁ To obtain intermediate B-1

Wherein, ring A, R ₁ 、R ₂ 、R ₄ As defined in claim 1;

R ₃ is C ₁ -C ₆ Carboxyl or C ₁ -C ₆ An amide group; preferably, R ₃ Is C ₁ -C ₆ A carboxyl group; more preferably, R ₃ Is butyric acid.

19. The method of claim 18, wherein step i) is performed under nitrogen blanket; and/or, reacting in the presence of a catalyst;

preferably, the catalyst is a rhodium catalyst and a copper catalyst;

more preferably, the catalyst is rhodium acetate.

20. The method as recited in claim 18, wherein said step i) further comprises the step of:

i-1) reaction of Compound B-3Substituent R of ₃ After being protected by a carboxyl protecting group, the intermediate B-1 reacts to obtain a compound shown in a formula I protected by the carboxyl protecting group;

i-2) removing the carboxyl protecting group in the compound shown in the formula I protected by the carboxyl protecting group to obtain the compound shown in the formula I.

21. A pharmaceutical composition, comprising: a compound according to any one of claims 1 to 8; and a pharmaceutically acceptable carrier.

22. The pharmaceutical composition of claim 21, further comprising at least one additional pharmacologically active inhibitor; preferably, the other pharmacologically active inhibitor comprises an antibody selected from CTLA4, PDL1 or PD 1.

23. Use of a compound according to any one of claims 1 to 8, or a pharmaceutical composition according to claim 21 or 22, for the prevention and/or treatment of inflammatory diseases, pain, cancer, metabolic diseases, diseases of the urinary system;

and/or for the preparation of a medicament, pharmaceutical composition or formulation for the prevention and/or treatment of inflammatory diseases, pain, cancer, metabolic diseases, urinary diseases.

24. The use of claim 23, wherein the inflammatory disease is selected from the group consisting of arthritis, rheumatoid arthritis; the pain is selected from osteoarthritis pain, pain caused by endometriosis; the cancer is selected from solid cancers, preferably, from breast cancer, cervical cancer, colorectal cancer, endometrial cancer, glioblastoma, head and neck cancer, kidney cancer, liver cancer, lung cancer, medulloblastoma, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer and urinary tract cancer; the metabolic disease is diabetes, and the urinary system disease is overactive bladder.

25. The use of claim 23, wherein the medicament is administered in combination with radiation therapy and/or antibody therapy, wherein the antibody therapy is selected from one or a combination of CTLA4 antibody therapy, PDL1 antibody therapy and PD1 antibody therapy.