Spiro compounds as inhibitors of indoleamine-2,3-dioxygenase

Abstract

The present application discloses an indoleamine-2,3-dioxygenase inhibitor and a preparation method thereof. The inhibitor of the present application has a structure as shown in general formula (I), wherein Ar, E, Y, X, V, D, W, B, ring A and ring B are defined in the description and claims. The present application also discloses a preparation method of the inhibitor. The compound of general formula (I) of the present application can be used as an indoleamine-2,3-dioxygenase inhibitor to prepare a medicine for preventing and / or treating an indoleamine-2,3-dioxygenase mediated disease.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and particularly relates to an IDO inhibitor containing a spiro structure and a preparation method thereof. BACKGROUND

[0002] Indoleamine-2,3-dioxygenase (IDO) is a monomeric enzyme containing ferroheme discovered for the first time in cells by Hayaishi group in 1967, the cDNA coding protein is composed of 403 amino acids, and the molecular weight is 45 kDa. It is a rate-limiting enzyme for the catabolism along the tryptophan-kynurenine pathway, and has a wide expression in various mammalian tissues. In the cells of tumor patients, IDO often plays an important physiological role in inducing tumor microenvironment immune tolerance, and the tryptophan (Trp)-kynurenine (Kyn) metabolic pathway mediated by IDO is involved in tumor immune escape, and IDO also plays an important role in inducing tumor microenvironment immune tolerance.

[0003] Tryptophan is one of the important essential amino acids in mammals, which needs to be taken in large amounts from food to maintain cell activation and proliferation and the synthesis of some neurotransmitters. Therefore, its deficiency will cause dysfunction of some important cells. IDO can catalyze the conversion of tryptophan to N-formylkynurenine in vivo, degrade the content of tryptophan and cause the deficiency of tryptophan in vivo, leading to the occurrence of tumors. Immunohistological studies have shown that the kynurenine pathway can lead to an increase in the excitotoxin quinolinic acid, and also cause various serious human diseases such as Alzheimer's disease and other nervous system diseases.

[0004] There are two main tryptophan rate-limiting enzymes in mammals: tryptophan dioxygenase (TDO) and IDO. In 1937, Kotake et al. purified the protein from rabbit intestine and first discovered that TDO is mainly expressed in the liver of mammals, and so far no close relationship between him and the immune system has been found. TDO can catalyze the kynurenine pathway to convert tryptophan to N-formylkynurenine. In 1978, the enzyme purified from the rabbit intestine was identified as a ferriheme-containing dioxygenase (IDO), which is the only enzyme that can catalyze the oxidation and cleavage of indole in the tryptophan molecule and catabolize along the kynurenine pathway outside the liver. IDO is usually expressed in organs with more mucous membranes, such as the lung, small intestine, large intestine, rectum, spleen, kidney, stomach and brain, and is widely distributed. Under certain special or pathological conditions, such as pregnancy, chronic infection, organ transplantation and tumors, the expression of IDO will significantly increase, and it is involved in mediating local immunosuppression.

[0005] Studies have shown that IDO can inhibit local T cell immune response in tumor microenvironment in the following ways: tryptophan depletion, toxic metabolism and induction of regulatory T cell proliferation. Many cases are overexpressed in tumors, thereby depleting local tryptophan and producing a large amount of kynurenine metabolites. In fact, under tryptophan-free or kynurenine-free culture conditions, T cells will have proliferation inhibition, activity reduction and even apoptosis. There is a very sensitive regulation point in T cells to tryptophan level, and under the action of IDO, tryptophan can be depleted, thereby causing T cells to be arrested in the middle of G1 phase, thereby inhibiting the proliferation of T cells and the immune response of T cells. Once the T cells stop proliferating, they may no longer be stimulated, which is the immune mechanism of IDO in vivo.

[0006] There is still a need in the art to develop new IDO inhibitors with high activity, and the present application finds that a new type of compound containing a spiro ring structure has unexpectedly high IDO inhibitory activity. SUMMARY

[0007] The purpose of the present application is to provide a new type of compound containing a spiro ring structure as an efficient IDO enzyme inhibitor.

[0008] Another purpose of the present application is to provide a preparation method of the compound.

[0009] In a first aspect, the present application provides a compound of general formula (I) or a stereoisomer or tautomer thereof, or a pharmaceutically acceptable salt or prodrug thereof:

[0010]

[0011] In the formula,

[0012] Ar is C6-C 20 aryl, C5-C 20 heteroaryl; Ar can be substituted with one or more groups selected from the group consisting of halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, hydroxyl, amino, nitro, aldehyde, -CF3, -CN, -SF5, NR a R b , carboxyl, -COR a , -CO2C1-C6alkyl, -CONR a R b , -SO2R e , -SO2NR a R b , -P(O)Me2, -P(O)(OMe)2; wherein each R a and each R b is independently hydrogen, substituted or unsubstituted C1-C 10alkyl, substituted or unsubstituted C3-C 10 cycloalkyl, substituted or unsubstituted C2-C 10 alkenyl, substituted or unsubstituted C6-C 20 aryl, or substituted or unsubstituted C3-C 14 heteroaryl, substituted or unsubstituted C1-C 10 alkylene C6-C 10 aryl, substituted or unsubstituted C1-C 10 alkylene C2-C 10 heteroaryl; R a and R b may together form a three- to eight-membered ring or a four- to eight-membered heterocyclic ring, wherein the heteroatom can be sulfur, oxygen, NH or NR b ;

[0013] E is a bond, -O-, -S-, -NR a -, -C(R a )= or -C(R a R b )2-;

[0014] Y is C(R 1 ), =C, N;

[0015] X is C(R 1 ), N;

[0016] R 1 is hydrogen, OH, OC1-C 10 alkyl, C1-C 10 alkyl;

[0017] Ring A and Ring B are connected to form a spiro structure;

[0018] Ring A and Ring B can each be a 3- to 12-membered carbocyclic ring; or

[0019] Ring A and Ring B can each be a 3- to 12-membered carbobicyclic ring, or

[0020] Ring A and Ring B can each be a 3- to 12-membered carbobridged bicyclic ring, or

[0021] Ring A and Ring B can each be a 3- to 12-membered carbocyclic ring, wherein one or more carbon ring atoms can be replaced by one or more O, S, -C(O)-, -C(S)-, NR b ;

[0022] Ring A and Ring B can each be a 3- to 12-membered carbocyclic ring, which is unsubstituted or can be substituted by one or more R c ; or

[0023] Ring A and Ring B can each be a 3- to 12-membered carbocyclic ring, wherein one carbon ring atom can be replaced by a nitrogen atom;

[0024] V is a bond or C1-C6alkylene; V can be substituted by one to three groups selected from the group consisting of C1-C6alkyl, OC1-C6alkyl, C3-C6cycloalkyl; or V is NR b or CR f R g ; R f and R g may together form a three- to eight-membered ring or a four- to eight-membered heterocyclic ring, wherein the heteroatoms can be sulfur, oxygen, NH or NR b ;

[0025] R f and R g are each hydrogen, C1-C6alkyl, C3-C6cycloalkyl, aryl, heteroaryl, C1-C6alkylenearyl, C1-C6alkyleneheteroaryl, C1-C6alkylenecycloalkyl; R f may be substituted by one or more substituents selected from the group consisting of C1-C6alkyl, OC1-C6alkyl or C3-C6cycloalkyl;

[0026] D is C(O), C(=NOH), C(S) or S(O)2;

[0027] W is a bond, -O-, -CR a R b - or -N(R 5 )-;

[0028] R 5 is hydrogen, C1-C6alkyl, aryl, heteroaryl;

[0029] B is C6-C 20 aryl, C5-C 20 heteroaryl; B can be substituted by one or more groups selected from the group consisting of halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, hydroxy, amino, nitro, aldehydo, -CF3, -CN, -SF5, NR a R b , carboxy, -COR a , -CO2C1-C6alkyl, -CONR a R b , -SO2R e , -SO2NR a R b , -P(O)Me2, -P(O)(OMe)2; wherein each R a and each R b is independently hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C 10 cycloalkyl, substituted or unsubstituted C2-C 10 alkenyl, substituted or unsubstituted C6-C 20 aryl, or substituted or unsubstituted C3-C 14 heteroaryl; R a and R b may together form a three- to eight-membered ring or a four- to eight-membered heterocyclic ring, wherein the heteroatom can be sulfur, oxygen, NH or NR b ;

[0030] R e is C1-C 10 alkyl, C3-C 10 cycloalkyl, C6-C 20 aryl, or C3-C 14 heteroaryl; R e may be substituted by one or more groups selected from the group consisting of halogen, hydroxyl, amino, nitro, cyano, aldehyde, carboxyl, alkoxy, -CF3, -SF5.

[0031] In another preferred embodiment, Ar is:

[0032]

[0033] wherein:

[0034] Z and T are independently CH, CR e or N;

[0035] R 2 , R 3 and R 4 are independently hydrogen, halogen, C1-C6haloalkyl, hydroxyl, amino, nitro, aldehyde, -CF3, -CN, -SF5, NR a R b , carboxyl, -COR a , -CO2C1-C6alkyl, -CONR a R b , -SO2R e , -SO2NR a R b , -P(O)Me2, -P(O)(OMe)2.

[0036] In another preferred embodiment, E in general formula (I) is a chemical bond or O.

[0037]

[0038] In another preferred embodiment, E in general formula (I) is a chemical bond or O.

[0039] In another preferred embodiment, Y in general formula (I) is CH.

[0040] In another preferred embodiment, X in general formula (I) is CH or N.

[0041] In another preferred embodiment, V in general formula (I) is a bond, -C(C1-C6alkyl)-, -N(R 5 ), or -N(CH2Ar 1 ); Ar 1 is substituted or unsubstituted phenyl; Ar 1 may be substituted by one or more radicals selected from the group consisting of halogen, C1-C6alkyl, C1-C6haloalkyl.

[0042] In another preferred embodiment, D in general formula (I) is -C(O)- or -C(=NOH)-.

[0043] In another preferred embodiment, W in general formula (I) is a bond or -N(R 5 ).

[0044] In another preferred embodiment, B in general formula (I) is substituted or unsubstituted phenyl; B can be substituted by one or more radicals selected from the group consisting of halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, hydroxyl, -CN, -SF5.

[0045] In another preferred embodiment, the compound of general formula (I) is according to general formula (II):

[0046]

[0047] wherein R 2 , ring A, ring B and B are defined as in claim 1, R 5 is hydrogen, C1-C6alkyl, OC1-C6alkyl, C3-C6cycloalkyl, C1-C6alkylenearyl; Z is O or NOH.

[0048] In another preferred embodiment, the compound of general formula (I) is according to general formula (III):

[0049]

[0050] wherein R 2 , ring A, ring B and B are defined as in claim 1, R 5 is hydrogen, C1-C6alkyl, C3-C6cycloalkyl, C1-C6alkylenearyl.

[0051] In another preferred embodiment, the compound of general formula (I) is according to general formula (IV):

[0052]

[0053] wherein R 2 , ring A, ring B and B are as defined in claim 1, R 5 , R 6 and R 7 are each independently hydrogen, C1-C6 alkyl, OC1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylene aryl; R 6 and R 7 may together form a three- to eight-membered ring or a four- to eight-membered heterocyclic ring, wherein the heteroatom can be sulfur, oxygen, NH or NR b ; n is an integer from 1 to 6; and Z is O or NOH.

[0054] In another preferred embodiment, the compound of general formula (I) is as shown in general formula (V):

[0055]

[0056] wherein R 2 , ring A, ring B and B are as defined in claim 1, R 5 , R 6 and R 7 are each independently hydrogen, C1-C6 alkyl, OC1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylene aryl; R 6 and R 7 may together form a three- to eight-membered ring or a four- to eight-membered heterocyclic ring, wherein the heteroatom can be sulfur, oxygen, NH or NR b ; n is an integer from 1 to 6; and Z is O or NOH.

[0057] In another preferred embodiment, the compound of general formula (I) is as shown in general formula (II) (VI),

[0058]

[0059] wherein R 2 , ring A, ring B and B are as defined in claim 1, R 6 is hydrogen, C1-C6 alkyl, OC1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylene aryl; and Z is O or NOH.

[0060] In another preferred embodiment, the compound of general formula (I) is as shown in general formula (VII),

[0061]

[0062] wherein R 2 , ring A, ring B and B are as defined in claim 1, R 6is hydrogen, C1-C6 alkyl, OC1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylene aryl; Z is O or NOH.

[0063] In another preferred embodiment, the compound of general formula (I) is as shown in general formula (VIII),

[0064]

[0065] Where R 2 is halogen, R 6 is hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylene aryl; Z is O or NOH; Ar 3 is substituted or unsubstituted phenyl, and the substituents may be selected from one or more halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, hydroxyl, amino, nitro, -CF3, -CN, -SF5, NR a R b , carboxyl, -COR a 、-CO2C1-C6 alkyl、-CONR a R b 、-SO2R e 、-SO2NR a R b 、-P(O)Me2、-P(O)(OMe)2,where R a and R b As defined in claim 1.

[0066] In another preferred embodiment, the compound of formula (I) is as shown in formula (IX),

[0067]

[0068] Where R 2 is halogen, R 6 is hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkylene aryl; Z is O or NOH; Ar 3 is substituted or unsubstituted phenyl, and the substituents may be selected from one or more halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, hydroxyl, amino, nitro, -CF3, -CN, -SF5, NR a R b , carboxyl, -COR a 、-CO2C1-C6 alkyl、-CONR a R b 、-SO2R e 、-SO2NR a R b 、-P(O)Me2、-P(O)(OMe)2,where Ra and R b as defined in claim 1.

[0069] In another preferred embodiment, the compound of general formula (I) to (IX) is:

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076] In another preferred embodiment, the stereoisomer is a cis-trans isomer.

[0077] In another preferred embodiment, the compound is a racemate.

[0078] In another preferred embodiment, the stereoisomer is an enantiomer.

[0079] In another preferred embodiment, any one or more hydrogens in the compound can be replaced by deuterium.

[0080] In another preferred embodiment, the pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, trifluoromethanesulfonate, benzenesulfonate, p-toluenesulfonate (tosylate), 1-naphthalenesulfonate, 2-naphthalenesulfonate, acetate, trifluoroacetate, malate, tartrate, citrate, lactate, oxalate, succinate, fumarate, maleate, benzoate, salicylate, phenylacetate, mandelate.

[0081] The compound of general formula (I) of the present application can be obtained by the following preparation method, comprising the following steps:

[0082]

[0083] wherein R is C1-C6 alkyl, C1-C6 haloalkyl; Ar 2 as defined for B; Ar 1 as defined for Ar, and other groups or atoms are as previously defined.

[0084]

[0085] wherein R is C1-C6 alkyl, C1-C6 haloalkyl; Ar 2B; Ar 1 B; Ar

[0086]

[0087] B; Ar 2 B; Ar 1 B; Ar

[0088]

[0089] B; Ar 2 B; Ar 1 B; Ar

[0090]

[0091] B; Ar 2 B; Ar 1 B; Ar

[0092]

[0093] B; Ar 2 B; Ar 1 B; Ar

[0094]

[0095] B; Ar 2 B; Ar 1 B; Ar

[0096]

[0097] B; Ar 2 B; Ar 1 B; Ar

[0098] In the above method,

[0099] The base can be selected from the group consisting of alkali metal hydroxide, alkaline earth metal hydroxide, alkali metal hydride, alkaline earth metal hydride, alkali metal salt of HMDS, pyridine, triethylamine and the like.

[0100] The acid can be selected from the group consisting of hydrochloric acid, sulfuric acid and the like.

[0101] The dehydrating agent can be selected from the group consisting of HATU and the like.

[0102] The palladium catalyst can be selected from the following group: tetrakistriphenylphosphine palladium and the like.

[0103] The catalyst can be selected from the following group: palladium carbon, etc.

[0104] The reducing agent can be selected from the following group: LiAlH4.

[0105] Another aspect of the present invention provides the use of the compound of formula (I) or its stereoisomers or tautomers, or its pharmaceutically acceptable salts or prodrugs according to the first aspect, for:

[0106] (i) preparing indoleamine-2,3-dioxygenase inhibitors;

[0107] (ii) preparing a medicament for preventing and / or treating an indoleamine-2,3-dioxygenase-mediated disease; or

[0108] (iii) preparing anti-inflammatory drugs.

[0109] In another preferred embodiment, the indoleamine-2,3-dioxygenase-mediated disease is a disease with pathological characteristics of the tryptophan metabolic pathway mediated by IDO.

[0110] In another preferred embodiment, the indoleamine-2,3-dioxygenase-mediated disease is cancer, eye disease, psychological disorder, depression, anxiety, Alzheimer's disease and / or autoimmune disease.

[0111] In another preferred embodiment, the cancer includes but is not limited to: colon cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, pancreatic cancer, ovarian cancer, prostate cancer, kidney cancer, liver cancer, brain cancer, melanoma, multiple myeloma, chronic myeloid leukemia, blood tumors, lymphomas, including metastatic lesions in other tissues or organs away from the primary site of the tumor.

[0112] Another aspect of the present invention provides a pharmaceutical composition comprising:

[0113] The compound of the general formula (I) according to the first aspect of the present invention or its stereoisomers or tautomers, or its pharmaceutically acceptable salts or prodrugs; and

[0114] Pharmaceutically acceptable carrier.

[0115] In another preferred embodiment, the pharmaceutical composition further contains other anti-tumor drugs.

[0116] In another preferred embodiment, the other anti-tumor drugs are selected from the following group: PD-1 antibody, PD-L1 antibody, CTLA-4 antibody and other anti-tumor chemotherapy drugs and targeted drugs.

[0117] In another preferred embodiment, the other anti-tumor drug includes, but is not limited to, an immunotherapeutic drug for cancer: PD-1 antibody, CTLA-4 antibody, PD-L1 antibody, PD-L2 antibody, any other chemotherapeutic drug or targeted therapeutic drug, such as HDAC inhibitor, arginine metabolic enzyme inhibitor, STIN activator and EP4 antagonist.

[0118] In another aspect of the present application, there is provided a method for preventing and / or treating an indoleamine-2,3-dioxygenase-mediated disease, comprising the step of administering to a patient a compound of general formula (I) as described above or a stereoisomer or a tautomer thereof, or a pharmaceutically acceptable salt or prodrug thereof, or a pharmaceutical composition as described above.

[0119] In another preferred embodiment, the indoleamine-2,3-dioxygenase-mediated disease is cancer, and the method further comprises the step of administering to the patient an additional anti-cancer agent (also referred to as an anti-tumor drug, which is described above).

[0120] The compound of general formula (I) of the present application has various pharmacological activities, such as anti-tumor, treating neurodegenerative diseases (Alzheimer's disease), anti-inflammatory, etc.

[0121] It should be understood that, within the scope of the present application, each of the technical features described above and each of the technical features specifically described below (e.g., in the examples) can be combined with each other to form a new or preferred technical solution. Due to the limited space, they will not be repeated one by one here. Specific embodiments

[0122] The present inventors have made extensive and in-depth research and for the first time accidentally developed a new compound containing a sulfoximine and a 1,2,5-oxadiazole structure. The compound can be used as a highly efficient IDO enzyme inhibitor for preventing and / or treating an indoleamine-2,3-dioxygenase-mediated disease, and can also be used as an anti-inflammatory drug. On this basis, the present application is completed.

[0123] Definitions

[0124] The term "C1-C 10 The term "C1-C" refers to a monovalent saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms, including straight and branched chain hydrocarbon groups, such as methyl (i.e., CH3-), ethyl (i.e., CH3CH2-), n-propyl (i.e., CH3CH2CH2-), isopropyl (i.e., (CH3)2CH-), n-butyl (i.e., CH3CH2CH2CH2-), isobutyl (i.e., (CH3)2CHCH2-), sec-butyl (i.e., (CH3)(CH3CH2)CH-), t-butyl (i.e., (CH3)3C-), n-pentyl (i.e., CH3CH2CH2CH2CH2-), neopentyl (i.e., (CH3)3CCH2-). In the present application, this term includes substituted or unsubstituted alkyl groups.

[0125] As used herein, the term "substituted or unsubstituted" means that the group can be unsubstituted or that the H of the group is replaced by one or more (preferably 1-6, more preferably 1-3) substituents.

[0126] As used herein, "substituted" or "substitution" means that the group has one or more (preferably 1-6, more preferably 1-3) substituents selected from the group consisting of halogen, hydroxy, -NH2, nitro, -CN, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, C2-C6alkenyl, C2-C6alkynyl, phenyl, benzyl, C1-C6alkylS(O)2-, (C0-C6alkyl)2NS(O)2-, C1-C6alkylC(O)-, C3-C6cycloalkylC(O)-, C0-C6alkylOC(O)-, (C0-C6alkyl)2NC(O)-, C0-C6alkylC(O)NH-, (C0-C6alkyl)2NC(O)NH-.

[0127] As used herein, the term "C3-C 12 Cycloalkyl" means a cyclic, substituted or unsubstituted, ring having 3 to 12 carbon atoms, such as -CH2-cyclopropane, -CH2-cyclobutane.

[0128] As used herein, the term "alkoxy" means -O-alkyl, wherein the alkyl group can be saturated or unsaturated, can be branched, straight-chained, or cyclic. Preferably, the alkoxy group has 1 to 10 carbon atoms, i.e. C1-C10alkoxy. More preferably, the alkoxy group has 1 to 6 carbon atoms, i.e. C1-C6alkoxy. Representative examples include, but are not limited to: methoxy, ethoxy, propoxy. 10 Alkoxy, preferably 1 to 6 carbon atoms. Representative examples include, but are not limited to: methoxy, ethoxy, propoxy.

[0129] As used herein, the term "C6-C 20 Aryl" means a monovalent aromatic carbocyclic group of from 6 to 20 (preferably 6 to 14) carbon atoms having a single ring (e.g., phenyl) or a conjugated dual ring, which is either hydrocarbocyclic (e.g., naphthyl) or heterocyclic (e.g., thienyl or furyl), which can be substituted or unsubstituted, wherein the substituents are defined above. Preferred aryl groups include phenyl and naphthyl. The term includes substituted or unsubstituted forms, wherein the substituents are defined above.

[0130] As used herein, the term "C2-C 10 Alkenyl" means an alkenyl group having from 2 to 10 (e.g., 2 to 6 or 2 to 4) carbon atoms and having at least one (e.g., 1 to 2) sites of aliphatic unsaturation (i.e., >C=C<). Examples of such groups include ethenyl, allyl, but-3-enyl.

[0131] As used herein, the term "C3-C 10 "Cycloalkyl" refers to a cyclic alkyl group having from 3 to 10 carbon atoms, having a single ring or multiple rings (including fused, bridged, and spiro ring systems). In a fused ring system, one or more rings may be cycloalkyl, heterocyclic, aryl, or heteroaryl, as long as the point of attachment is through the cycloalkyl ring. Examples of suitable cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclooctyl.

[0132] As used herein, the term "halo" or "halogen" refers to fluoro, chloro, bromo, and iodo.

[0133] As used herein, the term "heteroaryl" refers to an aromatic group having 1 to 10 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur in the ring. As for the term "heteroaryl" indicating the number of carbon atoms, such as "C3-C 20 "Heteroaryl" means an aromatic group having 3-20 carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfur. Other similar groups. Such heteroaryl groups may be monocyclic (such as pyridyl or furyl) or fused rings (such as indolizinyl or benzothienyl), wherein the fused ring may be non-aromatic and / or contain one heteroatom, as long as the point of attachment is through the atom of the aromatic heteroaryl group. In one embodiment, the ring atoms nitrogen and / or sulfur of the heteroaryl group are optionally oxidized to N-oxide (NO), sulfinyl or sulfonyl. Preferred heteroaryl groups include pyridyl, pyrrolyl, indolyl, thienyl and furyl. The term includes substituted or unsubstituted heteroaryl groups.

[0134] As used herein, the term "substituted heteroaryl" refers to a heteroaryl group substituted with 1 to 5, preferably 1 to 3, more preferably 1 to 2 substituents selected from the same substituents as defined for substituted aryl.

[0135] As used herein, the term "heterocycle" or "heterocyclic" or "heterocycloalkyl" or "heterocyclyl" refers to a saturated, partially saturated or unsaturated group (but not aromatic) having a single ring or fused rings (including bridged ring systems and spiro ring systems) with 1 to 10 carbon atoms and 1 to 4 (e.g., 3) heteroatoms selected from nitrogen, sulfur or oxygen within the ring. In a fused ring system, one or more rings can be cycloalkyl, aryl or heteroaryl, as long as the point of attachment is through the non-aromatic ring. In one embodiment, the nitrogen atom and / or sulfur atom of the heterocyclic group is optionally oxidized to provide N-oxide, sulfinyl and sulfonyl moieties.

[0136] As used herein, the term "substituted heterocyclic" or "substituted heterocycloalkyl" or "substituted heterocyclyl" refers to a heterocyclic group substituted with 1 to 5 (e.g., 1 to 3) substituents, the substituents being the same as those defined for substituted cycloalkyl.

[0137] As used herein, the term "stereoisomer" refers to a chiral different compound at one or more stereocenters. Stereoisomers include enantiomers and diastereomers.

[0138] As used herein, the term "tautomer" refers to an alternative form of a compound that differs in the position of a proton, such as enol-keto and imine-enamine tautomers, or tautomeric forms of heteroaryl groups that contain ring atoms connected to both an -NH- moiety of the ring and an =N- moiety of the ring, such as pyrazole, imidazole, benzimidazole, triazole, and tetrazole.

[0139] "Prodrug" refers to any derivative of a compound of the embodiments that, when administered to a subject, is able to provide directly or indirectly a compound of the embodiments or an active metabolite or residue thereof. Particularly preferred derivatives and prodrugs are those that enhance the bioavailability of the compounds of the embodiments when administered to a subject (e.g., by allowing an orally administered compound to be more readily absorbed into the blood) or that enhance transport of the parent compound to a biological compartment (e.g., the brain or lymphatic system) relative to the parent species. Prodrugs include ester forms of the compounds of the present invention.

[0140] Compounds of the present invention

[0141] As used herein, the term "compounds of the present invention" refers to compounds of general formula (I), racemates thereof, stereoisomers thereof or tautomers thereof, or prodrugs, or pharmaceutically acceptable salts thereof.

[0142] The present invention relates to: racemic mixtures of these compounds, mixtures enriched in either enantiomer, and any isolated enantiomer. For the scope of the present invention, it is understood that the racemic mixture refers to a 50%:50% mixture of the two R and S enantiomers. The isolated enantiomer is understood to be a pure enantiomer (i.e. 100%) or a mixture highly enriched in one enantiomer (purity > 98%, > 95%, > 93%, > 90%, > 88%, > 85%, > 80%).

[0143] In case the compounds according to the present invention exist as stereoisomers, the present invention comprises all stereoisomers of the compounds.

[0144] In case the compounds according to the present invention exist as tautomers, the present invention comprises all tautomers of the compounds.

[0145] The present invention also includes deuterated compounds, wherein any one or more hydrogen atoms in the compounds described are replaced by a stable isotope thereof, deuterium.

[0146] Pharmaceutical compositions

[0147] The present application also provides a pharmaceutical composition comprising a safe and effective amount of the active ingredient, and a pharmaceutically acceptable carrier.

[0148] The "active ingredient" of the present application refers to the compound of the general formula (I) or its stereoisomer or tautomer, or a pharmaceutically acceptable salt or prodrug thereof.

[0149] The "active ingredient" and the pharmaceutical composition of the present application can be used as an IDO inhibitor. In another preferred embodiment, for the preparation of a medicament for the prevention and / or treatment of a tumor. In another preferred embodiment, for the preparation of a medicament for the prevention and / or treatment of an IDO-mediated disease.

[0150] A "safe and effective amount" refers to the amount of the active ingredient sufficient to substantially improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the active ingredient per dose, more preferably, 10-200 mg of the active ingredient per dose. Preferably, the "dose" is a tablet.

[0151] A "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid filler diluents or gelling agents suitable for human use that must have sufficient purity and sufficiently low toxicity. "Compatible" in this context means that the components of the composition can be combined with the active ingredient of the present application and with each other, without any significant degradation of the activity of the active ingredient.

[0152] The compounds of the preferred embodiments can be administered as the sole active agent or in combination with one or more other agents useful in the treatment of cancer. The compounds of the preferred embodiments are also effective when used in combination with known therapeutic agents and anti-cancer agents. Combinations of the presently known compounds and other anticancer or chemotherapeutic agents are within the scope of the preferred embodiments. Examples of such agents can be found in Cancer Principles and Practice of Oncology by V.T. Devita and S. Hellman (editors), 6thedition (February 15, 2001), Lippincott Williams & Wilkins Publishers. Based upon the particular nature of the drug and the cancer involved, one of ordinary skill in the art can identify effective combinations of agents. Such anti-cancer agents include, but are not limited to, the following: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic / cytostatic agents, antiproliferative agents, prenyl-protein transferase inhibitors, histone deacetylase (HDAC) inhibitors, HMG-CoA reductase inhibitors and other angiogenesis inhibitors, cell proliferation and survival signal inhibitors, inducers of apoptosis, and agents that interfere with cell cycle checkpoints, CTLA4 antibodies, PD-1 antibodies, PD-L1 antibodies, and the like. The compounds of the preferred embodiments are also effective when administered concurrently with radiation therapy.

[0153] Generally, the compounds of the preferred embodiments will be administered in a therapeutically effective amount by any of the accepted modes of administration for agents having similar utilities. The actual amount of the compound of the preferred embodiments (i.e., active ingredient) will depend upon numerous factors such as the severity of the disease being treated, the age and relative health of the patient, the potency of the compound being used, the mode of administration and the bioavailability of the compound being used, and other factors well known in the art. The drug can be administered as frequently as necessary, such as once a day or twice a day. All of these factors are within the skill of the attending clinician.

[0154] The therapeutically effective dose of the preferred embodiments can generally be the total daily dose administered to the patient in a single dose or in divided doses, e.g., about 0.001 to about 1000 mg / kg body weight per day, preferably about 1.0 to about 30 mg / kg body weight per day. Dosage unit compositions can contain such amounts of the preferred embodiments that can be admixed with a carrier or vehicle so as to provide the desired dosage in a single dose or in divided doses. The selection of carrier or vehicle will be guided by a variety of factors, such as the mode of administration and the bioavailability of the drug substance. Generally, the compounds of the preferred embodiments can be administered as pharmaceutical compositions by any route of administration, including orally, systemically (e.g., transdermally, intranasally, or via a suppository), or parenterally (e.g., intramuscularly, intravenously, or subcutaneously). The preferred mode of administration is oral, and convenient daily doses can be adjusted to accommodate the degree of bitterness. The compositions can take the form of tablets, pills, capsules, semisolids, powders, sustained release formulations, solutions, suspensions, elixirs, aerosols, or any other appropriate compositions. Another preferred mode of administration of the compounds of the preferred embodiments is inhalation. This is an effective method of delivering a therapeutic agent directly to the respiratory tract (see, e.g., U.S. Patent No. 5,607,915).

[0155] Suitable pharmaceutically acceptable carriers or excipients include, for example, processing agents and drug delivery modifiers and facilitators, such as calcium phosphate, magnesium stearate, talc, monosaccharides, disaccharides, starch, gelatin, cellulose, sodium methylcellulose, carboxymethyl cellulose, dextrose, hydroxypropyl-B-cyclodextrin, polyvinylpyrrolidone, low melting waxes, ion exchange resins, and the like, and combinations of any two or more thereof. Liquid and semisolid excipients can be selected from glycerol, propylene glycol, water, ethanol, and various oils, including those of petroleum, animal, vegetable or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesame oil, etc. Preferred liquid carriers, particularly for injectable solutions, include water, saline, aqueous dextrose, and glycols. Additional suitable pharmaceutically acceptable excipients are described in Remington's Pharmaceutical Sciences, Mack Pub. Co., New Jersey (1991), incorporated herein by reference.

[0156] As used herein, the term "pharmaceutically acceptable salt" means a non-toxic acid or base salt of a compound of Formula (I). These salts can be prepared in situ during the final isolation and purification of the compounds of Formula (I), or separately by reacting the appropriate organic or inorganic acid or base with the basic or acidic functionalities of the compound, respectively. Representative salts include, but are not limited to: acetate, adipate, alginate, citrate, aspartate, benzoate, besylate, bisulfate, butyrate, camphorate, camphorsulfonate, di gluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, palmoate, pectinate, thiocyanate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, sulfate, tartrate, thiocyanate, tosylate, and undecanoate. Additionally, the nitrogen-containing basic groups can be quaternized with agents such as alkyl halides, such as methyl, ethyl, propyl, butyl, phenyl and benzyl chlorides, bromides and iodides; dialkyl sulfates, such as dimethyl, diethyl, dibutyl and diamyl sulfates; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides, such as benzyl and phenethyl bromides; and others. The resulting water or oil-soluble or dispersible products can be thereby obtained. Examples of acids which can be employed to form a pharmaceutically acceptable acid addition salt include, for example, inorganic acids such as hydrochloric, sulfuric, phosphoric, and the like, and organic acids such as oxalic, maleic, methanesulfonic, succinic, citric, and the like. Base addition salts can be prepared during the final isolation and purification of the compounds of Formula I, or separately, by reacting the carboxyl moiety with a suitable base such as the hydroxides, carbonate or bicarbonate of a pharmaceutically acceptable metal cation such as sodium, potassium, calcium, magnesium, aluminum, and the like, or with ammonia, or an organic primary, secondary or tertiary amine. Pharmaceutically acceptable salts include, but are not limited to, salts of alkali and alkaline earth metal cations such as sodium, lithium, potassium, calcium, magnesium, aluminum, and the like, as well as nontoxic ammonium, quaternary ammonium, and amine cations including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. Other representative organic amines useful for the formation of base addition salts include diethylamine, ethanolamine, diethanolamine, piperazine, and the like.

[0157] As used herein the term "pharmaceutically acceptable prodrug" refers to those prodrugs of the compounds of the preferred embodiments which are rapidly transformed in vivo to the parent compound of the above formula, for example, by hydrolysis in blood. A thorough discussion is provided in "T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S. 15 Symposium Series," and in "Edward B. Roche, Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987," both of which are incorporated herein by reference.

[0158] The present application has the advantages of:

[0159] (1) providing a novel compound of the general formula (I);

[0160] (2) the compound of the present application can be used as an efficient IDO enzyme inhibitor;

[0161] (3) the synthesis method is mild, simple and easy to operate, has high yield, is easy to derivatize, and is suitable for industrial mass production;

[0162] (4) has various pharmacological activities such as anti-tumor, neurodegenerative disease (Alzheimer's disease), anti-inflammatory, etc.

[0163] The present application will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and are not used to limit the scope of the present application. The experimental methods in the following examples are not specified, and are generally carried out according to the conventional conditions, such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight.

[0164] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. In addition, any method and material similar or equivalent to those described herein can be used in the practice of the present application. The described preferred methods and materials are only examples.

[0165] Example 1

[0166] (±)N-(4-chlorophenyl)-6-(quinolin-4-yl)spiro[3.3]heptane-2-carboxamide

[0167]

[0168] First Step: (3,3-dimethoxycyclobutane-1,1 -diyl)dimethanol

[0169]

[0170] Lithium aluminum hydride (28.81 g, 759.7 mmol) was added portionwise to a three-necked flask containing tetrahydrofuran (1500 mL), then a solution of isopropyl 3,3-dimethoxycyclobutane-1,1 -dicarboxylate (99.57 g, 345.3 mmol) in tetrahydrofuran (500 mL) was added dropwise slowly. Stirring at room temperature for 12 h, TLC showed the reaction was complete. Quenching by dropwise addition of saturated potassium sodium tartrate solution (27 mL) under ice bath, stirring at room temperature for 1 h. Filtration under suction, the filter cake was washed with dichloromethane / methanol (10:1, 200 mL x 5), the organic filtrate was combined and rotary evaporation to give the product as colorless transparent oil (59.8 g, yield 95%).

[0171] 1 H NMR (400 MHz, CDC13): δ 1.94 (s, 4H), 3.12 (s, 6H), 3.70 (s, 4H).

[0172] Second Step: (3,3-dimethoxycyclobutane-1,1 -diyl)bis(methylene)bis(4- methylbenzenesulfonate)

[0173]

[0174] The product of the first step (56.7 g, 321.8 mmol) was dissolved in pyridine (500 mL), p-toluenesulfonyl chloride (153.4 g, 804.7 mmol) was added portionwise under ice bath, stirring at room temperature for 12 h, TLC showed the reaction was complete. The reaction was filtered, the filtrate was slowly poured into 1000 mL water, solid precipitated, filtration under suction to give white solid (137.3 g, yield 88%).

[0175] 1 H NMR (400 MHz, CDC13): δ 1.92 (s, 4H), 2.45 (s, 6H), 3.01 (s, 6H), 3.99 (s, 4H), 7.36 (d, 4H), 7.74 (d, 4H).

[0176] Third Step: Diisopropyl 6,6-dimethoxyspiro[3.3]heptane-2,2-dicarboxylate

[0177]

[0178] In a three-necked flask was added N,N-dimethylformamide (300 mL), sodium hydride (9.63 g, 240.8 mmol) was added portionwise, purged with nitrogen three times, then diisopropyl propiolate (41.61 g, 221.1 mmol) was added dropwise, stirred at room temperature for 1 h. A solution of the second step product (53.03 g, 109.4 mmol) and potassium iodide (1.82 g, 10.94 mmol) in N,N-dimethylformamide (200 mL) was added to the reaction system. Heating was started, and the reaction was carried out at 140 °C for 12 h. TLC showed that the reaction was complete. After the reaction was cooled, it was poured into 1000 mL of water, extracted with petroleum ether (500 mL x 3), rotary evaporated, and distilled under reduced pressure (125-127 °C / 2 mmHg) to obtain the product as a transparent yellowish oil (16.9 g, yield 47%).

[0179] 1 H NMR (400 MHz, CDC13): δ 1.22 (d, 12H), 2.21 (s, 4H), 2.58 (s, 4H), 3.12 (s, 6H), 5.04 (sep, 2H).

[0180] Fourth step: Diisopropyl 6-oxospiro[3.3]heptane-2,2-dicarboxylate

[0181]

[0182] The third step product (1.00 g, 3.04 mmol) was added to a round-bottom flask, 3 mol / L hydrochloric acid (14 mL) was added, stirred at room temperature for 6 h, the reaction was complete, and white precipitate was generated. Filtration gave white precipitate (800 mg, yield 93%).

[0183] 1 H NMR (400 MHz, CDC13): δ 1.22 (d, 12H), 2.21 (s, 4H), 2.58 (s, 4H), 3.12 (s, 6H), 5.04 (sep, 2H).

[0184] Fifth step: Diisopropyl 6-(((trifluoromethyl)sulfonyl)oxy)spiro[3.3]hept-5-ene-2,2-dicarboxylate

[0185]

[0186] The product of the fourth step (700 mg, 2.48 mmol) was added to a three-necked flask, dry tetrahydrofuran was added, replaced with nitrogen for 3 times, cooled to -78 °C with dry ice acetone bath, 2 mol / L sodium bis(trimethylsilyl)amide tetrahydrofuran solution (1.49 mL, 2.98 mmol) was added dropwise slowly at -78 °C under nitrogen protection, stirred at -78 °C under nitrogen protection for 0.5 h. N-phenyl bis(trifluoromethanesulfonylimide) solution (1.06 g dissolved in 15 mL tetrahydrofuran) was added dropwise slowly, continued to stir at -78 °C under nitrogen protection for 0.5 h, the reaction was complete, saturated ammonium chloride solution (10 mL) was added to quench, 10 mL x 3 ethyl acetate was extracted for 3 times, the organic phase was combined, washed with saturated brine (10 mL), dried over sodium sulfate, filtered, the filtrate was concentrated, and column chromatography (petroleum ether: ethyl acetate = 80:1-40:1) was performed to obtain the target product (333 mg, 32.4%) as colorless oil.

[0187] 1 H NMR (400 MHz, CDCl3): δ 5.51 (s, 1H), 5.07 (sep, 2H), 2.92 (s, 2H), 2.75 (s, 4H), 1.25 (d, 6H), 1.24 (d, 6H).

[0188] Step 6: 6-(quinolin-4-yl)spiro[3.3]hept-5-ene-2,2-dicarboxylic acid diisopropyl ester

[0189]

[0190] The product of the fifth step (270 mg, 0.65 mmol), 4-quinoline boronic acid (135 mg, 0.78 mmol), potassium carbonate (180 mg, 1.3 mmol) were added to a flask, 5 mL dioxane, tetrakis(triphenylphosphine)palladium (75 mg, 0.065 mmol) were added, replaced with nitrogen for 3 times, the reaction was heated to 75 °C, stirred at 75 °C under nitrogen protection for 30 h, the reaction was complete. The reaction was used directly for the next step without treatment.

[0191] MS ESI: m / z = 394.1, [M+H] + .

[0192] Step 7: 6-(quinolin-4-yl)spiro[3.3]heptane-2,2-dicarboxylic acid diisopropyl ester

[0193]

[0194] To the reaction solution of the sixth step, 10% palladium-carbon (27 mg) and methanol (10 mL) were added, and the mixture was stirred at room temperature under normal pressure hydrogen for 3 h. The reaction was complete. The reaction solution was filtered, and the filtrate was concentrated and passed through a column (dichloromethane:methanol = 100:1-70:1) to obtain the target product (150 mg, total yield of two steps 58.3%) as a yellow oil.

[0195] MS ESI: m / z = 396.1, [M+H] + .

[0196] 1 H NMR (400 MHz, CDC13): δ 8.84 (d, 1H), 8.10-8.12 (m, 1H), 7.87-7.89 (m, 1H), 7.68-7.72 (m, 1H), 7.52-7.55 (m, 1H), 7.21 (d, 1H), 5.07 (sep, 2H), 3.96-4.04 (m, 1H), 2.83 (s 2H), 2.66-2.71 (m, 2H), 2.52 (s, 2H), 2.31-2.36 (m, 2H), 1.25 (t, 12H).

[0197] Eighth step: 6-(quinolin-4-yl)spiro[3.3]heptane-2,2-dicarboxylic acid

[0198]

[0199] The product of the seventh step (55 mg, 0.14 mmol) was added to a flask, and ethanol (10 mL), 2 mol / L sodium hydroxide (0.35 mL) were added. The reaction solution was heated to 85°C and reacted for 3 h. The reaction was complete. 2 mol / L hydrochloric acid was added to pH = 3, and the crude product (70 mg) was obtained by concentration.

[0200] MS ESI: m / z = 312.0, [M+H] + .

[0201] Ninth step: (±) 6-(quinolin-4-yl)spiro[3.3]heptane-2-carboxylic acid

[0202]

[0203] The crude product of the eighth step (70 mg) was added to a flask, and pyridine (10 mL) was added. The reaction solution was heated to reflux and reacted for 4 h. The reaction was complete. 2 mol / L hydrochloric acid was added to pH = 3, and the crude product (80 mg) was obtained by concentration.

[0204] MS ESI: m / z = 268.1, [M+H] + .

[0205] Tenth step: (±) N-(4-chlorophenyl)-6-(quinolin-4-yl)spiro[3.3]heptane-2- carboxamide

[0206]

[0207] The crude product of the ninth step (80 mg), triethylamine (0.076 mL, 0.56 mmol), N,N-dimethylformamide (5 mL) were added to a flask, 2-(7-oxobenzotriazol- yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (128 mg, 0.336 mmol) was added, stirred at room temperature for 30 min, p-chloroaniline (42 mg, 0.336 mmol) was added, stirred at room temperature for 32 h, the reaction was complete. Ethyl acetate (15 mL) was added, washed with water three times with 50 mL each time, the aqueous phase was extracted with ethyl acetate (3 x 10 mL) three times, the organic phase was combined, washed with saturated brine (5 mL), dried over sodium sulfate, filtered, the filtrate was concentrated, and columned (dichloromethane:methanol = 100:1-70:1) to give the crude product (30 mg), preparative TLC separation gave the yellow solid target product (12.2 mg, 23.1% overall yield for three steps).

[0208] MS ESI: m / z = 377.1, [M+H] + . 1 H NMR (400 MHz, CDC13): δ 8.84 (s, 1H), 8.11-8.13 (m, 1H), 7.89-7.91 (m, 1H), 7.68-7.72 (m, 1H), 7.50-7.56 (m, 4H), 7.22-7.28 (m, 2H), 3.97-4.06 (m, 2H), 3.05-3.11 (m, 1H), 2.04-2.69 (m, 8H).

[0209] Example 2

[0210] (±)(cis / trans) N-(4-chlorophenyl)-6-(quinolin-4-yl)spiro[2.5]octane-l- carboxamide

[0211]

[0212] First step: ethyl 2-(l,4-dioxaspiro[4.5]dec-8-ylidene)acetate

[0213]

[0214] Sodium hydride (52 mg, 0.88 mmol) and triethyl phosphonoacetate (52 mg, 0.83 mmol) were dissolved in N,N-dimethylformamide 5 mL, stirred for half an hour, then compound 1,4-dioxaspiro[4.5]decan-8-one (100 mg, 0.65 mmol) dissolved in 2 mL N,N-dimethylformamide was added dropwise into the reaction solution, and stirring was continued at room temperature for 1 hour. 50 mL of water was added for quenching, and ethyl acetate (20 mL x 3) was used for extraction. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and rotary evaporated to obtain a white oil (128 mg, 88%).

[0215] 1 H NMR (400 MHz, CDCl3): δ 5.60 (s, 1H), 4.05-4.11 (m, 2H), 3.91-3.92 (d, 4H), 2.95 (t, 3H), 2.31 (t, 3H), 1.67-1.73 (m, 4H), 1.19-1.23 (m, 3H).

[0216] Second step: (±) Ethyl 7,10-dioxaspiro[2.2.4 6 .2 3 ]dodecan-1-carboxylate

[0217]

[0218] Trimethylsulfonium iodide (193 mg, 0.80 mmol) and potassium tert-butoxide (99 mg, 0.88 mmol) were dissolved in dimethyl sulfoxide 5 mL, stirred at room temperature for half an hour, and the product of the first step (60 mg, 0.27 mmol) dissolved in 1 mL dimethyl sulfoxide was added. Stirring was continued at room temperature for 2 days, and the reaction was quenched with water (50 mL). Ethyl acetate (20 mL x 3) was used for extraction, and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and rotary evaporated to obtain an oil (46 mg, 43%).

[0219] MS ESI: m / z = 241.1, [M+H] + .

[0220] Third step: (±) Ethyl 6-oxospiro[2.5]octane-1-carboxylate

[0221]

[0222] The second step product (100 mg, 0.42 mmol) was dissolved in tetrahydrofuran 5 mL, then 3 mol / L HCl (5 mL) was added and stirred overnight, the buffer was quenched (pH = 6, 50 mL), extracted with ethyl acetate (20 mL x 3), the organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, rotary evaporated, and columned (PE:EA = 4:1) to obtain a white oil (86 mg, 105%).

[0223] MS ESI: m / z = 197.1, [M+H] + .

[0224] Fourth step: (±) ethyl 6-(((trifluoromethyl)sulfonyl)oxy)spiro[2.5]oct-5-ene-1-carboxylate

[0225]

[0226] Under nitrogen protection, -78 °C, N-phenyl-bis(trifluoromethylsulfonyl)imide (85 mg, 0.43 mmol), sodium bis(trimethylsilyl)amide (0.26 mL, 2 mmol / L 0.52 mmol) were dissolved in tetrahydrofuran (3 mL), stirred for half an hour, the third step product (186 mg, 0.52 mmol) was added and kept at low temperature for 4 hours, quenched with a buffer salt, added a phosphate buffer solution (pH = 6, 50 mL), extracted with ethyl acetate (30 mL x 3), the organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, rotary evaporated, and columned (PE:EA = 5:1) to obtain a white oil (93 mg, 65%).

[0227] 1 H NMR (400 MHz, CDCl3): δ 5.69 (t, 1H), 4.03-4.10 (m, 2H), 2.28-2.20 (m, 3H), 1.83-1.97 (m, 3H), 1.51-1.62 (m, 1H), 1.12-1.22 (m, 4H), 0.90-0.94 (m, 1H).

[0228] Fifth step: (±) ethyl 6-(quinolin-4-yl)spiro[2.5]oct-5-ene-1-carboxylate

[0229]

[0230] The product of Step 4 (90 mg, 0.28 mmol), quinoline-4-boronic acid (62 mg, 0.36 mmol), were dissolved in tetrahydrofuran (15 mL), potassium carbonate (114 mg, 0.82 mmol) was added, and a catalytic amount of tetrakis(triphenylphosphine)palladium (42 mg, 10%) was added. The reaction was stirred at 95 °C under nitrogen for 10 h. The catalyst was filtered off, and the reaction was quenched with a buffer solution (pH = 6, 100 mL). The product was extracted with ethyl acetate (50 mL x 3), and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (PE:EA = 5:1) to give a yellow solid (30 mg, 36%).

[0231] MS ESI: m / z = 308.1, [M+H] + .

[0232] Step 6: (±) (cis / trans) 6-(quinolin-4-yl)spiro[2.5]octane-1-carboxylic acid ethyl ester

[0233]

[0234] The product of Step 5 (460 mg, 1.5 mmol) was dissolved in ethanol (5 mL), and palladium on carbon catalyst (46 mg, 10%) was added. The reaction was stirred under hydrogen overnight. The palladium on carbon was filtered off, and the solvent was concentrated. The residue was purified by column chromatography (PE:EA = 5:1) to give a white solid (450 mg, 97%).

[0235] MS ESI: m / z = 310.1, [M+H] + .

[0236] Step 7: (±) (cis / trans) 6-(quinolin-4-yl)spiro[2.5]octane-1-carboxylic acid

[0237]

[0238] The product of Step 6 (30 mg, 0.1 mmol) was dissolved in tetrahydrofuran / ethanol (2 mL / 2 mL), and 1 mol / L lithium hydroxide (2 mmol, 2 mL) was added. The reaction was stirred at 70 °C for 8 h. The reaction was quenched with a phosphate buffer solution (pH = 6, 100 mL). The product was extracted with ethyl acetate (50 mL x 3), and the combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography to give two isomers: isomer A (15 mg) and isomer B (10 mg).

[0239] MS ESI: m / z = 282.1, [M+H] + .

[0240] Step 8: (±) (cis / trans) N-(4-chlorophenyl)-6-(quinolin-4-yl)spiro[2.5]octane-1- carboxamide

[0241]

[0242] The isomer A (15 mg, 0.053 mmol) obtained in the seventh step, 4-chloroaniline (8.2 mg, 0.064 mmol), 2-(7-oxobenzotriazolyl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (24 mg, 0.064 mmol), diisopropylethylamine (28 μL, 0.16 mmol) were dissolved in dichloromethane (5 mL) and stirred at room temperature for 2 hours. The reaction was quenched by adding phosphate buffered saline solution (pH = 6, 100 mL), extracted with ethyl acetate (50 mL x 3), the organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by column (PE:EA = 1:1) to give compound 2A as a white solid (8 mg, 39%).

[0243] MS ESI: m / z = 391.1, [M+H] + .

[0244] In the same manner, the isomer B obtained in the seventh step gave compound 2B as a white solid (9 mg, 44%).

[0245] MS ESI: m / z = 391.1, [M+H] + .

[0246] Example 3

[0247] (±)N-(4-chlorophenyl)-6-(6-fluoroquinolin-4-yl)spiro[3.3]heptane-2-carboxamide

[0248]

[0249] First step: 6-(6-fluoroquinolin-4-yl)-6-hydroxyspiro[3.3]heptane-2,2-dicarboxylic acid diisopropyl ester

[0250]

[0251] To a solution of the product from the third step in Example 1 (0.5708 g, 2.022 mmol) in 6 mL of dry tetrahydrofuran was added tert-butyllithium (2.50 mL of 1.6 M in pentane, 4.0 mmol) dropwise at -78 °C under argon. After 3 minutes, 4-bromo-6-fluoroquinoline (0.4539 g, 2.008 mmol) in 20 mL of dry tetrahydrofuran was added dropwise. After stirring for 1 hour, the reaction was allowed to warm to room temperature, acidified with 0.14 mL of acetic acid, concentrated in vacuo, and purified by column (PE:EA = 0:100-2:3) to give the target product as a yellow oil (0.43 g, 50% yield).

[0252] MS ESI: m / z = 430.2, [M+H] + .

[0253] Second step: (±) 6-(6-fluoroquinolin-4-yl)spiro[3.3]heptane-2-carboxylic acid

[0254]

[0255] The second step product (1.48 g, 3.45 mmol), red phosphorus (0.55 g, 17.8 mmol) were mixed in 10 mL of 55% concentrated hydroiodic acid, stirred at 140 °C for 4 hours, then raised to 180 °C for 20 hours, cooled, filtered to remove red phosphorus, added 2.25 g of sodium carbonate to neutralize the acid, added 1.0 g of sodium thiosulfate pentahydrate to remove elemental iodine, then added 6.50 g of sodium dihydrogen phosphate dihydrate to adjust the pH, extracted three times with ethyl acetate (40 + 20 + 20 mL), combined the organic phase, washed the organic phase with saturated brine, and dried the organic phase with anhydrous sodium sulfate. The organic phase was concentrated and columned (DCM:MeOH = 0:100-100:4) to obtain a white solid (1.48 g, yield 43%).

[0256] MS ESI: m / z = 286.1, [M+H] + .

[0257] Third step: (±) N-(4-chlorophenyl)-6-(6-fluoroquinolin-4-yl)spiro[3.3]heptane-2- carboxamide

[0258]

[0259] The ninth step crude product (80 mg), triethylamine (0.076 mL, 0.56 mmol), N,N- dimethylformamide (5 mL) were added to a flask, 2-(7-azabenzotriazol-1-yl)-1,1,3,3- tetramethyluronium hexafluorophosphate (128 mg, 0.336 mmol) was added, stirred at room temperature for 30 minutes, p-chloroaniline (42 mg, 0.336 mmol) was added, stirred at room temperature for 32 hours, the reaction was complete. Added ethyl acetate (15 mL), washed three times with 50 mL of water each time, the aqueous phase was extracted three times with ethyl acetate (3 x 10 mL), the organic phases were combined, washed with saturated brine (5 mL), dried with sodium sulfate, filtered, and the filtrate was concentrated and columned (dichloromethane:methanol = 100:1-70:1) to obtain a crude product (30 mg), which was separated by preparative TLC to obtain a yellow solid target product (12.2 mg, total yield of three steps 23.1%).

[0260] MS ESI: m / z = 377.1, [M+H] + .

[0261] 1 H NMR (400 MHz, CDC13): δ 8.84 (s, 1H), 8.11-8.13 (m, 1H), 7.89-7.91 (m, 1H), 7.68-7.72 (m, 1H), 7.50-7.56 (m, 4H), 7.22-7.28 (m, 2H), 3.97-4.06 (m, 2H), 3.05-3.11 (m, 1H), 2.04-2.69 (m, 8H).

[0262] Example 4

[0263] (±)(cis / trans) N-(4-chlorophenyl)-6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-1- carboxamide

[0264]

[0265] First step: 1,4-dioxaspiro[4.5]dec-7-en-8-yl trifluoromethyl sulfonate

[0266]

[0267] Into a 500 mL two-necked flask, 1,4-cyclohexanedione monoethylene ketal (18.44 g, 118.07 mmol) and N-phenyl bis(trifluoromethylsulfonimide) (46.4 g, 129.88 mmol) were added, and super dry tetrahydrofuran (200 mL) was added. The reaction flask was protected by N2, and cooled to -78 °C in dry ice acetone bath. Then 2 mol / L sodium bis(trimethylsilyl)amide tetrahydrofuran solution (71 mL, 141.68 mmol) was added slowly dropwise into the reaction flask, and the dropwise addition was completed in 45 min. After 1 h, TLC showed that the starting material disappeared, and the reaction was completed. Saturated ammonium chloride solution (15 mL) was added to quench the reaction, and the reaction solution was dried by rotary evaporation. Ethyl acetate (300 mL) was added, and the solution was washed twice with 5% sodium hydroxide solution (250 mL). The ethyl acetate phase was dried, and the product (27 g, yield: 80%) was obtained by rotary evaporation without further treatment, and was directly used in the next step.

[0268] 1 NMR (400 MHz, CDC13): δ 5.66 (t, 1H), 3.99 (d, 4H), 2.54 (s, 2H), 2.40 (s, 2H), 1.91 (t, 2H).

[0269] Second step: 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)-1,3,2- dioxaborolane

[0270]

[0271] To a two necked flask containing 1,4-dioxane (300 mL) was added the first step product (26 g, 83.84 mmol), bis(pinacolato)diboron (24.5 g, 96.43 mmol), potassium acetate (24.69 g, 251.53 mmol), sodium bromide (3.45 g, 33.53 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (3.42 g, 4.19 mmol), N2 protection, heated to 105 °C for 8 h. TLC showed the starting material was consumed. The reaction was concentrated, ethyl acetate (300 mL) was added, the insoluble material was filtered. Column chromatography on silica gel (petroleum ether: ethyl acetate, 10:0 to 8:2) gave the product as a light yellow oil (15 g, yield 68%).

[0272] 1 H NMR (400 MHz, CDC13): δ 6.45 (m, 1H), 3.96 (s, 4H), 2.36-2.33 (m, 4H), 1.73-1.70 (t, 2H), 1.23 (s, 12H).

[0273] Third step: 6-fluoro-4-(1,4-dioxaspiro[4.5]dec-7-en-8-yl)quinoline

[0274]

[0275] To a two necked flask containing 1,4-dioxane (300 mL) was added the first step product (26 g, 83.84 mmol), bis(pinacolato)diboron (24.5 g, 96.43 mmol), potassium acetate (24.69 g, 251.53 mmol), sodium bromide (3.45 g, 33.53 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (3.42 g, 4.19 mmol), N2 protection, heated to 105 °C for 8 h. TLC showed the starting material was consumed. The reaction was concentrated, ethyl acetate (300 mL) was added, the insoluble material was filtered. Column chromatography on silica gel (petroleum ether: ethyl acetate, 10:0 to 8:2) gave the product as a light yellow oil (15 g, yield 68%).

[0276] MS ESI: m / z = 286.1, [M+H] + .

[0277] 1H NMR (400 MHz, CDC13): δ 8.81-8.80 (d, 1H), 8.12-8.08 (q, 1H), 7.64-7.62 (q, 1H), 7.49-7.44 (td, 1H), 7.27-7.23 (d, 1H), 5.76-5.74 (m, 1H), 4.08-4.07 (m, 4H), 2.64-2.60 (m, 2H), 2.55-2.54 (m, 2H), 2.01-1.98 (t, 2H).

[0278] Fourth Step: 6-Fluoro-4-(l,4-dioxaspiro[4.5]dec-8-yl)quinoline

[0279]

[0280] The third step product (13 g, 45.61 mmol) was dissolved in a two-neck flask containing isopropanol (130 mL), 10% palladium carbon (1.3 g) was added, and the mixture was stirred at 55 °C under an atmosphere of hydrogen for 16 h. LCMS showed that the molecular weight of the reaction raw material disappeared, and the reaction was completed. The reaction solution was filtered and concentrated to obtain the crude product 10 g, which was directly used in the next step without further treatment.

[0281] MS ESI: m / z = 288.1, [M+H] + .

[0282] Fifth Step: 4-(6-Fluoroquinolin-4-yl)cyclohexanone

[0283]

[0284] The fourth step product (10 g, 34.84 mmol) was dissolved in a single-neck flask containing acetone (100 mL), then 4 mol / L hydrochloric acid (30 mL) was added, and the reaction was heated to 45 °C. After 3 h, TLC monitoring showed that the raw material disappeared, and the reaction was completed. The reaction solution was concentrated, ethyl acetate (100 mL) and water (100 mL) were added, and the pH was adjusted to 9 with saturated aqueous sodium bicarbonate solution, the aqueous phase was separated, and the aqueous phase was extracted with ethyl acetate (50 mL*2), the ethyl acetate phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography with petroleum ether: ethyl acetate (10:0-3:7) to obtain the product 5.7 g (yield: 74.1%).

[0285] MS ESI: m / z = 244.1, [M+H] + .

[0286] Sixth Step: (±) Ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohexylidene)acetate

[0287]

[0288] To sodium hydride (55 mg, 1.37 mmol) was added 5 mL DMF, then a solution of phosphoacetic acid triethyl ester (0.258 mL, 1.3 mmol) in 1 mL DMF was added dropwise, a solution of the product from Step 5 (0.243 g, 1.00 mmol) in 1 mL DMF was added, and the reaction was stirred at room temperature for 1 h. The DMF was evaporated, 100 mL of phosphate buffer (pH = 6) was added, and the mixture was extracted with ethyl acetate (3 x 50 mL). The organic layers were combined, washed with saturated brine (50 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 100:0 to 1:1) to give the desired product.

[0289] MS ESI: m / z = 314.1, [M+H] + .

[0290] Step 7: (±) (cis / trans) Ethyl 6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-1- carboxylate

[0291]

[0292] Potassium tert-butoxide (0.097 g, 0.44 mmol) and trimethylsulfonium iodide (0.049 g, 0.44 mmol) were dissolved in 5 mL DMSO and stirred at room temperature overnight. Then a solution of the product from Step 6 (0.069 g, 0.22 mmol) in 1 mL DMSO was added, and the reaction was stirred at room temperature for 2 days. The reaction was quenched by the addition of 200 mL water, and the mixture was extracted with ethyl acetate (3 x 50 mL). The organic layers were combined, washed with saturated brine (50 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 100:0 to 5:1) to give the desired product.

[0293] MS ESI: m / z = 328.2, [M+H] + .

[0294] Step 8: (±) (cis / trans) 6-(6-Fluoroquinolin-4-yl)spiro[2.5]octane-1-carboxylic acid

[0295]

[0296] The product of Step 6 (0.077 g, 0.23 mmol) was dissolved in 5 mL of ethanol at room temperature, and a solution of LiOH (0.198 g, 4.7 mmol) in 1 mL of water was added. The reaction was heated at 70 °C for 5 h, quenched with 200 mL of water, extracted with ethyl acetate (3 x 50 mL), the organic layers combined, washed with saturated brine (50 mL), dried over sodium sulfate, filtered, and the filtrate concentrated and purified by column (petroleum ether: ethyl acetate = 100:0 to 1:1) to give the target product.

[0297] MS ESI: m / z = 300.2, [M+H] + .

[0298] Step 9: (±)(cis / trans) N-(4-chlorophenyl) 6-(6-fluoroquinolin-4-yl) spiro[2.5]octane-1- carboxamide

[0299]

[0300] Following the procedure of Example 2, the reagent of Step 8 (±)(cis / trans) 6-(quinolin-4- yl) spiro[2.5]octane-1-carboxylic acid was replaced with (±)(cis / trans) 6-(6-fluoroquinolin-4- yl) spiro[2.5]octane-1-carboxylic acid, and the same conditions were used to give the target compound.

[0301] MS ESI: m / z = 409.1, [M+H] + .

[0302] Example 5

[0303] (±)(cis / trans) N-(4-fluorophenyl) 6-(6-fluoroquinolin-4-yl) spiro[2.5]octane-1-carboxamide

[0304]

[0305] Following the procedure of Example 4, the reagent of Step 9 4-chloroaniline was replaced with 4-fluoroaniline, and the same conditions were used to give the target compound.

[0306] MS ESI: m / z = 393.2, [M+H] + .

[0307] Example 6

[0308] (±)(cis / trans) 6-(6-fluoroquinolin-4-yl)-N-(4-(trifluoromethyl)phenyl) spiro[2.5]octane-1- carboxamide

[0309]

[0310] The reagent of step nine, 4-chloroaniline, was replaced with 4-aminobenzotrifluoride according to the procedure of Example 4 and the same conditions to give the target compound.

[0311] MS ESI: m / z = 443.2, [M+H] + .

[0312] Example 7

[0313] (±) (cis / trans) 6-(6-fluoroquinolin-4-yl)-N-phenylspiro[2.5]octane-l- carboxamide

[0314]

[0315] The reagent of step nine, 4-chloroaniline, was replaced with aniline according to the procedure of Example 4 and the same conditions to give the target compound.

[0316] MS ESI: m / z = 375.1, [M+H] + .

[0317] Example 8

[0318] (±) (cis / trans) 4-chloro-N-(6-(6-fluoroquinolin-4-yl)spiro[2.5]oct-l- yl)benzamide

[0319]

[0320] First step: (±) (cis / trans) 6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-l- amine

[0321]

[0322] The product of Example 4, step eight (20 mg, 0.067 mmol), was dissolved in toluene with diphenylphosphoryl azide (144 uL, 0.670 mmol), triethylamine (19 uL, 0.13 mmol) under a nitrogen atmosphere and heated at 70 °C for three hours. The toluene was evaporated and water / tetrahydrofuran (0.4 mL / 0.4 mL) was added followed by lithium hydroxide (15 mg, 0.64 mmol) and stirred at room temperature for 30 minutes. This was extracted three times with ethyl acetate (3 x 50 mL) and the organic phases were combined, washed with saturated brine (50 mL), dried over sodium sulfate, filtered and the filtrate was concentrated to give the target compound.

[0323] MS ESI: m / z = 270.1, [M+H] + .

[0324] Second step: (±) (cis / trans) 4-chloro-N-(6-(6-fluoroquinolin-4-yl)spiro[2.5]oct-l- yl)benzamide

[0325]

[0326] Following the procedure of Example 2, the reagent of Step 8, (±) (cis / trans) 6- (quinolin-4-yl)spiro[2.5]octane-l-carboxylic acid, p-chloroaniline was replaced with p-chlorobenzoic acid, the product of Step 1, and the same conditions were used to obtain the target compound.

[0327] MS ESI: m / z = 409.1, [M+H] + .

[0328] Example 9

[0329] (±) 1 -(4-chlorophenyl)-3-(6-(6-fluoroquinolin-4-yl)spiro[2.5]oct- 1 - yl)urea

[0330]

[0331] The product of Step 1 of Example 8 (20 mg, 0.074 mmol) and triethylamine (26 uL, 0.15 mmol) were dissolved in 3 mL of dichloromethane under nitrogen atmosphere, p-chlorophenyl isocyanate (14 mg, 0.088 mmol) in 1 mL of dichloromethane was added and the reaction was allowed to proceed at room temperature for 1 hour. Water (20 mL) was added and the mixture was extracted with ethyl acetate (50 mL*3), the organic phase was combined and washed with saturated brine (50 mL), dried over sodium sulfate, filtered and the filtrate was concentrated and purified by column (petroleum ether: ethyl acetate = 100:0-1 :1) to obtain the target compound.

[0332] MS ESI: m / z = 424.1, [M+H] + .

[0333] Example 10A and 10B

[0334] (±) N-(4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane- 1 -carboxamide

[0335] (±) N-(4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane- 1 -carboxamide

[0336]

[0337] Step 1 : 4-(4-cyclopropylidenecyclohexyl) 6-fluoroquinoline

[0338]

[0339] (3-bromopropyl)triphenylphosphonium bromide (5.72 g, 12.34 mmol) and sodium hydride (592.2 mg, 24.68 mmol) were placed in a two-necked flask under N2protection, then super dry tetrahydrofuran (15 mL) was injected into the reaction flask, heated to 70 °C for 4 hours. Subsequently, the product of example 4, step 5 (2 g, 8.23 mmol) dissolved in super dry tetrahydrofuran (15 mL) was injected into the reaction solution. After 2 hours, TLC monitoring showed that the starting material disappeared and the reaction was completed. The reaction solution was cooled to room temperature, the insoluble was filtered, and concentrated. Column chromatography with petroleum ether: ethyl acetate (10:0-8:2) gave the product as a white solid (1.4 g, yield: 63.6%).

[0340] MS ESI: m / z = 268.1, [M+H] + .

[0341] Second step: (syn / anti) 7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1-one

[0342]

[0343] MCPBA (370 mg, 2.15 mmol) was added to the product of the first step (500 mg, 1.87 mmol) in dichloromethane (5 mL) under ice bath, and stirred for 45 minutes under ice bath. TLC showed that the starting material disappeared and the intermediate was obtained. Then, methanesulfonic acid (360 mg, 3.74 mmol) was added to the reaction solution, and stirred for 2 hours at room temperature. TLC showed that the intermediate disappeared and the reaction was completed. Adjusted to pH = 9 with saturated aqueous NaHCO3solution, separated, the organic phase was washed with saturated aqueous sodium chloride solution (10 mL*2), dried over anhydrous sodium sulfate, and concentrated. Column chromatography with petroleum ether: ethyl acetate (100:0-75:25) gave the product syn isomer A-1 (160 mg) and anti isomer B-1 (200 mg) (yield: 67.9%). MS ESI: m / z = 284.1, [M+H] + .

[0344] Third step: (±) (syn / anti) 7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1-one

[0345]

[0346] Potassium tert-butoxide (297.4 mg, 2.65 mmol) was added to a solution of the product of the third step, cis / trans isomer A-1 (250 mg, 0.88 mmol), p-toluenesulfonylmethyl isocyanide (344.9 mg, 1.77 mmol) and methanol (42 mg, 1.31 mmol) in ethylene glycol dimethyl ether (10 mL) under ice bath conditions, stirred for 10 minutes, then returned to room temperature conditions to react. After 4 hours, the reaction was treated with saturated aqueous sodium bicarbonate solution (10 mL), separated, the aqueous phase was extracted with ethylene glycol dimethyl ether (10 mL), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and column chromatographed with petroleum ether: ethyl acetate (10:0-6:4) to obtain the product, cis / trans isomer A-2 (160 mg, yield: 61%).

[0347] MS ESI: m / z = 295.1, [M+H] + .

[0348] The preparation method of cis / trans isomer B-2 was the same as that of cis / trans isomer A-2, using the starting material cis / trans isomer B-1 instead of cis / trans isomer A-1 to obtain the product (116 mg, yield 37.29%).

[0349] MS ESI: m / z = 295.1, [M+H] + .

[0350] Fourth step: (±) (cis / trans) 7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-1-carboxylic acid

[0351]

[0352] The product of the third step, A-2 (150 mg, 0.54 mmol), was added to a 48% aqueous hydrobromic acid solution (4 mL), heated to 105°C for two days. LCMS monitoring showed that the starting material and intermediates disappeared, and the reaction was complete. It was neutralized to pH = 4 with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate (15 mL*3), the aqueous phase was dried over anhydrous sodium sulfate, and the ethyl acetate phase was concentrated to obtain the product, cis / trans isomer A-3 (147 mg, yield: 92.1%).

[0353] MS ESI: m / z = 314.1, [M+H] + .

[0354] The preparation method of cis / trans isomer B-3 was the same as that of cis / trans isomer A-3, using the starting material cis / trans isomer B-2 instead of cis / trans isomer A-2 to obtain the product (116 mg, yield 37.3%).

[0355] MS ESI: m / z = 314.1, [M+H] +.

[0356] Fifth step:

[0357] 10A (±) N-(4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-1- carboxamide

[0358]

[0359] The fourth step product syn / anti isomers A-3 (18.0 mg, 0.057 mmol), 4-chloroaniline (8.94 mg, 0.069 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (26.2 mg, 0.069 mmol) and N,N-diisopropylethylamine (22.3 mg, 0.172 mmol) were added to dichloromethane (2 mL), stirred at room temperature for 1 hour, then heated to 40 °C for 3 hours, LCMS showed the reaction was completed. Added water (10 mL), extracted with dichloromethane (10 mL*3), combined organic phase, dried with anhydrous sodium sulfate, column chromatography with petroleum ether: ethyl acetate (10:0-6:4) to get the product (9.06 mg, yield: 37.3%).

[0360] MS ESI: m / z = 423.1, [M+H] + .

[0361] 10B (±) N-(4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-1- carboxamide

[0362]

[0363] The fourth step product B-3 (12 mg, 0.038 mmol), 4-chloroaniline (5.9 mg, 0.046 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (17.5 mg, 0.046 mmol) and N,N-diisopropylethylamine (14.86 mg, 0.145 mmol) were added to dichloromethane (2 mL), stirred at room temperature for 1 hour, then heated to 40 °C for 3 hours, LCMS showed the reaction was completed. Added water (10 mL), extracted with dichloromethane (10 mL*3), combined organic phase, dried with anhydrous sodium sulfate, column chromatography with petroleum ether: ethyl acetate (10:0-6:4) to get the product 10B (5.76 mg, yield: 35.6%).

[0364] MS ESI: m / z = 423.1, [M+H] + .

[0365] Examples 11A and 11B

[0366] (±) 1 -(4-chlorophenyl)-3-(7-(6-fluoroquinolin-4-yl)spiro[3.5]non-1 - yl)urea

[0367] (±) 1 -(4-chlorophenyl)-3-(7-(6-fluoroquinolin-4-yl)spiro[3.5]non-1 - yl)urea

[0368]

[0369] First step: (±) 7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1 -amine

[0370]

[0371] The product of the fourth step in example 10 (20 mg, 0.064 mmol), diphenyl phosphorazide (21 mg, 0.077 mmol) and triethylamine (7.74 mg, 0.077 mmol) in toluene (0.6 ml) were heated to 70 °C under N2atmosphere. After two hours, the heating was stopped, the mixture was cooled to room temperature, the toluene was evaporated, water / tetrahydrofuran (0.4 mL / 0.4 mL), lithium hydroxide (15 mg, 0.64 mmol) were added and the mixture was stirred at room temperature for 30 minutes. The reaction was evaporated, ethyl acetate (20 mL) was added, stirred for 10 minutes, filtered, the ethyl acetate phase was dried over anhydrous sodium sulfate and concentrated to give the crude target product.

[0372] MS ESI: m / z = 285.2, [M+H] + .

[0373] Second step: (±) 1 -(4-chlorophenyl)-3-(7-(6-fluoroquinolin-4-yl)spiro[3.5]non-1 - yl)urea

[0374]

[0375] The product of the first step (15 mg, 0.053 mmol) and p-chlorophenyl isocyanate (8.9 mg, 0.058 mmol) were added to tetrahydrofuran (1 mL) and stirred at room temperature. After 30 minutes, TLC monitoring showed the disappearance of the starting material and the reaction was finished. Water (10 mL) was added, the mixture was extracted with ethyl acetate (10 mL*2), the ethyl acetate phase was dried, concentrated and purified by column chromatography with petroleum ether: ethyl acetate (10:0-6:4) to give the product (12.4 mg, yield: 53.9%).

[0376] MS ESI: m / z = 438.2, [M+H] + .

[0377] 11B (±) 1 -(4-chlorophenyl)-3-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan- 1 -yl)urea

[0378]

[0379] Following the procedure of Example 11A, the product of Example 10, syn-anti isomer B3, was used in place of the product of Example 10, syn-anti isomer A3, in the first step reaction to give the first step product, and the same reagent and procedure were used in the second step to give 11B (15.34 mg, yield: 66.7%). MS ESI: m / z = 438.2, [M+H] + .

[0380] Example 12A and 12B

[0381] (±) 4-chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1-yl)benzamide

[0382] (±) 4-chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1-yl)benzamide

[0383]

[0384] 12A (±) 4-chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1-yl)benzamide

[0385]

[0386] The product of Example 11, first step (15 mg, 0.053 mmol) and triethylamine (16 mg, 0.158 mmol) were dissolved in tetrahydrofuran (2 mL) under ice-bath conditions, and then p-chlorobenzoyl chloride (11.09 mg, 0.063 mmol) was added to the reaction, which was stirred at room temperature. After 15 minutes, TLC showed that the starting material was consumed, and the reaction was completed. Subsequently, water (10 mL) was added to the reaction, which was extracted with ethyl acetate (10 mL*3), and the combined organic phase was concentrated and purified by column chromatography with petroleum ether: ethyl acetate (10:0-6:4) to give the product as a white solid (14.58 mg, yield: 65.7%).

[0387] MS ESI: m / z = 423.1, [M+H] + .

[0388] 12B

[0389] (±) 4-chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1-yl)benzamide

[0390]

[0391] The product from the first step of example 11B was used instead of the product from the first step of example 11A following the procedure of example 12A to give example 12B (9.16 mg, 41.11% yield) under the same conditions.

[0392] MS ESI: m / z = 423.1, [M+H] + .

[0393] Examples 13A and 13B

[0394] (±) 4-Chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]non-1-yl)benzenesulfonamide

[0395] (±) 4-Chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]non-1-yl)benzenesulfonamide

[0396]

[0397] 13A

[0398] (±) 4-Chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]non-1-yl)benzenesulfonamide

[0399]

[0400] The product from the first step of example 11A (13.6 mg, 0.0479 mmol), 4- dimethylaminopyridine (0.58 mg, 0.005 mmol) and triethylamine (14.5 mg, 0.16 mmol) were dissolved in tetrahydrofuran (2 mL) and 4-chlorobenzenesulfonyl chloride (11.1 mg, 0.144 mmol) was added to the reaction under ice bath conditions. The reaction was stirred at room temperature overnight and TLC monitoring showed the disappearance of the starting material and the reaction was complete. Subsequently, water (10 mL) was added to the reaction and extracted with ethyl acetate (10 mL*3), the organic phases were combined, concentrated and purified by column chromatography with petroleum ether: ethyl acetate (10:0-6:4) to give the product as a white solid (11.8 mg, 53.8% yield).

[0401] MS ESI: m / z = 423.1, [M+H] + .

[0402] 13B

[0403] (±) 4-Chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]non-1-yl)benzenesulfonamide

[0404]

[0405] Prepared according to the procedure of Example 13A, replacing the first step product of Example 11A with the first step product of Example 11B, under the same conditions to give Example 13B (9.71 mg, 44.2% yield).

[0406] MS ESI: m / z = 459.1, [M+H] + .

[0407] Example 14A and 14B

[0408] (±)N-(3-bromophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1 -carboxamide

[0409] (±)N-(3-bromophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1 -carboxamide

[0410]

[0411] 14A (±)N-(3-bromophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1 -carboxamide

[0412]

[0413] Prepared according to the procedure of Example 10A, replacing p-chloroaniline with 3-bromoaniline under the same conditions to give Example 14A (12.15 mg, 81.6% yield).

[0414] MS ESI: m / z = 467.1, [M+H] + .

[0415] 14B (±)N-(3-bromophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-1 -carboxamide

[0416]

[0417] Prepared according to the procedure of Example 10B, replacing 4-chloroaniline with 3-bromoaniline under the same conditions to give Example 14B (6.89 mg, 46.3% yield).

[0418] MS ESI: m / z = 467.1, [M+H] + .

[0419] Example 15A and 15B

[0420] N-(4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0421] N-(4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0422]

[0423] First step: 8-methylene-1,4-dioxaspiro[4.5]decane

[0424]

[0425] In a three-necked flask was added methyltriphenylphosphonium bromide (13.0 g, 83.2 mmol), then diethyl ether (250 mL), potassium tert-butoxide (16.6 g, 147.7 mmol) was added portionwise under ice-bath, replaced nitrogen 3 times, left to room temperature under stirring for 1 h, 1,4-cyclohexanedione monoethylene ketal in diethyl ether (30 mL) was added dropwise slowly at 0 °C, removed ice-bath and heated to reflux for 8 h, TLC monitored the reaction was complete, filtered, the filtrate was dried under vacuum, extracted with ethyl acetate (100 mL*2), combined organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered off the sodium sulfate solid, the filtrate was dried under vacuum to give brown oil (10.38 g, yield 80%).

[0426] 1 H NMR (400 MHz, CDC13): δ 1.70 (t, 4H), 2.28 (t, 4H), 3.97 (s, 4H), 4.67 (s, 2H).

[0427] Second step: 8,1 1 -dioxaspiro[3.2.4 7 .2 4 ]tridecan-2-one

[0428]

[0429] In a three-necked flask was added the product of the first step (9.7 g, 62.9 mmol) and zinc-copper alloy (55.7 g, 869.0 mmol), added methyl tert-butyl ether (240 mL), replaced nitrogen 3 times, added a solution of trichloroacetyl chloride in ethylene glycol dimethyl ether (60 mL) dropwise slowly under ice-bath, left to room temperature under stirring, the reaction was left overnight. TLC monitored the reaction was complete, added saturated solution of ammonium chloride in methanol (300 mL) under ice-bath, left to room temperature under stirring for 4 h, filtered off the silica gel, the filtrate was dried under vacuum, column chromatography separation and purification (Flash: 0-20% ethyl acetate / n-hexane in), gave white solid (8.9 g, yield 73%).

[0430] 1H NMR (400 MHz, CDC13): δ 1.66 (t, 4H), 1.82 (t, 4H), 2.79 (s, 4H), 3.96 (s, 4H).

[0431] Third Step: 8, 11-Dioxaspiro[3.2.4 7 .2 4 ]tridecan-2-carbonitrile

[0432]

[0433] In a three-necked flask, add the product of the second step (5.0 g, 25.48 mmol), toluyl methyl isocyanide (10.44 g, 53.5 mmol), and anhydrous ethanol (1.8 g, 40.0 mmol), then add tert-butyl alcohol (50 mL), ethylene glycol dimethyl ether (50 mL), after stirring for 2 minutes, add potassium tert-butoxide (12.50 g, 112.1 mmol) under ice bath, and stir naturally back to room temperature, and react overnight. Monitor the reaction by TLC, after the reaction is complete, pour the reaction directly into ice water (50 mL), extract with ethyl acetate (50 mL*2), combine the organic phases, then wash with saturated brine (50 mL), and dry with anhydrous sodium sulfate. Filter off the sodium sulfate solid, and rotary evaporate the filtrate, and purify by column chromatography (Flash: 0-20% ethyl acetate / n-hexane) to obtain a white solid (2.4 g, yield 46%). 1 H NMR (400 MHz, CDC13): δ 1.66 (t, 4H), 1.82 (t, 4H), 2.79 (s, 4H), 3.96 (s, 4H).

[0434] Fourth Step: 7-Oxospiro[3.5]nonane-2-carbonitrile

[0435]

[0436] Add the product of the third step (6.88 g, 33.1 mmol) to a round-bottom flask, add acetonitrile and water (25 mL-25 mL), heat to an external temperature of 65 degrees, and stir for 3 hours. Monitor the reaction by TLC, add water (25 mL) when the reaction is complete, evaporate the acetonitrile from the reaction system, extract with ethyl acetate (25 mL*2), combine the organic phases, wash with saturated brine (25 mL), dry with anhydrous sodium sulfate, and purify by column chromatography (Flash: 0-60% ethyl acetate / n-hexane) (5.0 g, yield 92%).

[0437] 1H NMR (400 MHz, CDC13): δ 3.17-3.11 (m, 1H), 2.44-2.35 (m, 3H), 2.33-2.30 (m, 5H), 2.02-1.99 (m, 2H), 1.94-1.91 (m, 2H).

[0438] Fifth step: 2-cyano spiro[3.5]non-6-ene-7-yl trifluoromethanesulfonate

[0439]

[0440] The product of the fourth step (7.0 g, 42.89 mmol) was added to a three-necked flask, dry tetrahydrofuran was added, replaced with nitrogen for 3 times, cooled to -78 °C with dry ice acetone bath, slowly added 2 mol / L sodium bis(trimethylsilyl)amide tetrahydrofuran solution (25.73 mL, 51.46 mmol) at -78 °C under nitrogen protection, stirred for 1 h at -78 °C under nitrogen protection. Slowly added N-phenyl bis(trifluoromethanesulfonylimide) solution (18.40 g dissolved in 60 mL tetrahydrofuran), continue to stir for 1 h at -78 °C under nitrogen protection, the reaction was complete, added saturated ammonium chloride solution (10 mL) to quench, extracted with 50 mL x 3 ethyl acetate for 3 times, the organic phase was combined, washed with saturated brine (50 mL), dried over sodium sulfate, filtered, the filtrate was concentrated, and column chromatography (Flash: 0-20% ethyl acetate / n-hexane) was performed to obtain the target product as a colorless oil (12.0 g, yield 95%).

[0441] 1 H NMR (400 MHz, CDC13): δ 5.67-5.65 (m, 1H), 3.15-3.05 (m, 1H), 2.39-2.36 (m, 3H), 2.30-2.22 (m, 5H), 1.92-1.89 (t, 1H), 1.83-1.80 (t, 1H).

[0442] Sixth step: 7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[3.5]non-6-ene-2-carbonitrile

[0443]

[0444] The fifth step product (12.0 g, 40.64 mmol), pinacolboronate (12.38 g, 48.77 mmol), potassium acetate (11.97 g, 121.92 mmol) were added to a flask, replaced with argon 3 times, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(ll) (75 mg, 0.065 mmol) was added, replaced with nitrogen 3 times, the reaction was heated to 105 °C, and reacted at this temperature for 16 hours, the reaction was complete. The reaction was filtered through celite, the filtrate was concentrated, and column chromatography (Flash: 0-15% ethyl acetate / n-hexane) was used to purify to obtain the target compound (10.8 g, yield 97.3%).

[0445] 1 H NMR (400 MHz, CDC13): δ 6.43-6.41 (m, 1H), 3.06-3.03 (m, 1H), 2.25-2.14 (m, 8H), 1.68-1.65 (m, 1H), 1.61-1.58 (m, 1H), 1.26 (s, 6H), 1.25 (s, 6H).

[0446] Seventh step: 7-(6-fluoroquinolin-4-yl)spiro[3.5]non-6-ene-2-carbonitrile

[0447]

[0448] The sixth step product (10.8 g, 39.53 mmol), 4-bromo-6-fluoroquinoline (8.94 g, 39.53 mmol), tetrakis triphenylphosphine palladium (2.28 g, 1.98 mmol), and cesium carbonate (25.76 g, 79.07 mmol) were added to dioxane (120 mL) and water (30 mL), heated to an external temperature of 100 degrees, and reacted overnight, and TLC was used to monitor the completion of the reaction. The reaction was filtered, the filtrate was concentrated, and column chromatography (Flash: 0-30% ethyl acetate / n-hexane) was used to purify to obtain a white solid.

[0449] MS ESI: m / z = 293.2, [M+H] + .

[0450] 1 H NMR (400 MHz, CDC13): δ 8.81 (d, 1H), 8.12-8.09 (m, 1H), 7.51-7.45 (m, 2H), 7.16 (s, 1H), 5.75-5.74 (m, 1H), 3.23-3.15 (m, 1H), 2.43-2.36 (m, 8H), 1.99-1.96 (m, 2H).

[0451] Eighth Step: (±) 7-(6-Fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxylic acid

[0452]

[0453] The product of the seventh step (11.9 g, 40.73 mmol) was added to a flask, water (300 mL), isopropanol (2 mL), potassium hydroxide (45.6 g, 814 mmol) were added, the reaction was heated to 123 °C, and reacted for 16 h. The reaction was complete. 12 mol / L hydrochloric acid was added to acidify to pH = 6.5. Ethanol (150 mL) was added to the original reaction, replaced with nitrogen once, and palladium on carbon (1.0 g) was added. The reaction was carried out under a hydrogen atmosphere for 16 h. After the reaction was complete, the methanol and isopropanol in the reaction system were evaporated, and white solids were precipitated. The solution was cooled in the refrigerator for 10 min, and white solids were obtained by filtration (7.0 g, yield 80%).

[0454] MS ESI: m / z = 314.1, [M+H] + .

[0455] Ninth Step: (±) N-(4-Chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2- carboxamide

[0456]

[0457] Racemate 15: The product of the eighth step (150 mg, 0.48 mmol), 4-chloroaniline (58 mg, 0.48 mol), and 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (218 mg, 0.57 mmol) were added to N,N-dimethylformamide (3.0 mL), and N,N-diisopropylethylamine (185 mg, 1.44 mmol) was added while stirring in an ice bath. The solution was allowed to return to room temperature and stirred for 16 h. The reaction solution was added to water (10 mL), and then ethyl acetate (30 mL) was added. The organic phase was washed with saturated sodium bicarbonate solution (20 mL*2), saturated brine (20 mL), and anhydrous sodium sulfate. After filtration, the solvent was evaporated, and column chromatography (Flash: 0-60% ethyl acetate / n-hexane) was used for purification to obtain racemate 15 (100 mg, yield 50%).

[0458] Preparation of isomer A of the ninth step: 10.0 mg of racemate 15 was separated by chiral column AD-H (n-hexane: ethanol, 80:20) to obtain 15A (5.0 mg, yield 50%) with a retention time of 33 min.

[0459] Ninth step preparation of isomer B: racemic 15 take 10.0 mg, split by chiral column AD-H (n-hexane: ethanol, 80:20), the retention time of 39 minutes is 15B (5.0 mg, yield 50%).

[0460] Resolution of chiral isomers

[0461] Chiral resolution was performed using Agilent 1260 semi-preparative liquid chromatograph (Chiralpak).

[0462] Resolution of the compound of example 15:

[0463] Chiral column: CHIRALPAK AD-H 10*250 mm; flow rate 3.0 mL / min; detection wavelength 254 nm; collect two enantiomers.

[0464] Conditions: 10 mg of sample of example 15 was dissolved in 6 mL of methanol, the injection volume was 0.5 mL, the eluent was n-hexane: ethanol = 80:20 (volume ratio); the peak times of the two optical isomers were 33.0 minutes and 39.0 minutes, respectively.

[0465] Example 16

[0466] (±) 1-(4-chlorophenyl)-3-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-2-yl)urea

[0467]

[0468] The product of the eighth step of example 15 (30 mg, 0.096 mmol), diphenyl phosphinic azide (31.6 mg, 0.115 mmol) and triethylamine (37 mg, 0.288 mmol) were dissolved in toluene (0.5 mL), protected by N2, reacted at 70°C for 2 hours. Then p-chloroaniline (37 mg, 0.288 mmol) was added and reacted for 10 minutes. The reaction was then treated and directly column chromatographed to obtain the target product 9 mg, yield 21.4%, with petroleum ether: ethyl acetate (10:0-3:2).

[0469] MS ESI: m / z = 438.1, [M+H] + .

[0470] Example 17

[0471] (±) 4-chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-2-yl)benzamide

[0472]

[0473] First step: (±) 7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-2-amine

[0474]

[0475] The product of the eighth step of Example 15 (30 mg, 0.096 mmol), diphenyl phosphinic azide (26 mg, 0.096 mmol) and triethylamine (11.63 mg, 0.115 mmol) were dissolved in toluene (1 mL) and the reaction was carried out at 70 °C for 2 hours under N2protection. The toluene was then evaporated, 1 mol / L HC1 / THF (2 mL / 1 mL) was added and stirred at room temperature for 30 min. Saturated aqueous NaHC03solution (10 mL) was added and stirred for 10 min. The ethyl acetate phase was extracted with ethyl acetate (15 mL*3) and dried. The product was used directly in the next step without further purification.

[0476] MS ESI: m / z = 285.1, [M+H] + .

[0477] Second step: (±) 4-chloro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-2- yl)benzamide

[0478]

[0479] The product of the first step (15 mg, 0.0528 mmol) and triethylamine (16 mg, 0.158 mmol) were dissolved in tetrahydrofuran (2 mL). p-Chlorobenzoyl chloride (11.09 mg, 0.0634 mmol) was added to the reaction under ice bath condition and stirred for 30 min. LCMS monitoring showed the disappearance of the starting material and the reaction was completed. Water (10 mL) was added and the ethyl acetate phase was extracted with ethyl acetate (10 mL*3), dried, concentrated and then separated by preparative plate (petroleum ether: ethyl acetate = 2: 1) to give the product 3.55 mg (R f = 0.2).

[0480] MS ESI: m / z = 423.1, [M+H] + .

[0481] Example 18

[0482] (±) N-(4-bromophenyl)-7-(fluoroquinolin-4-yl)spiro[3.5]nonan-2- formamide

[0483]

[0484] The product of the ninth step was prepared according to the procedure of Example 15 by substituting p-bromoaniline for p-chloroaniline under the same conditions to give the target compound 24 mg, yield: 82%.

[0485] MS ESI: m / z = 467.0, [M+H] + .

[0486] Example 19

[0487] (±) N-(4-Fluorophenyl)-7-(fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0488]

[0489] Following the preparation method of example 15, the reagent p-chloroaniline of the ninth step was replaced by p-fluoroaniline, using the same conditions to give the target compound 21 mg, yield: 72%.

[0490] MS ESI: m / z = 467.0, [M+H] + .

[0491] Example 20

[0492] (±) 4-Bromo-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-2-yl)benzamide

[0493]

[0494] Following the preparation method of example 17, the reagent p-chlorobenzoyl chloride of the second step was replaced by p-bromobenzoyl chloride, using the same conditions to give the target compound 6.2 mg.

[0495] MS ESI: m / z = 467.0, [M+H] + .

[0496] Example 21

[0497] (±) 4-Fluoro-N-(7-(6-fluoroquinolin-4-yl)spiro[3.5]nonan-2-yl)benzamide

[0498]

[0499] Following the preparation method of example 17, the reagent p-chlorobenzoyl chloride of the second step was replaced by p-fluorobenzoyl chloride, using the same conditions to give the target compound 6.04 mg.

[0500] MS ESI: m / z = 467.0, [M+H] + .

[0501] Examples 22A and 22B

[0502] N-(4-Chlorophenyl)-2-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-7-carboxamide

[0503] N-(4-chlorophenyl)-2-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-7-carboxamide

[0504]

[0505] First step: 2-(6-fluoroquinolin-4-yl)-8,11-dioxaspiro[3.2.4 7 .2 4 ]tridecan-2-ol

[0506]

[0507] To a solution of the product of the first step (1.00 g, 5.1 mmol) in 50 mL of dry tetrahydrofuran was added tert-butyllithium (6.38 mL, 1.6 mol / L in pentane, 10.2 mmol) dropwise at -78°C under argon atmosphere. After 3 minutes, a solution of 4-bromo-6-fluoroquinoline (1.15 g, 5.1 mmol) in 30 mL of dry tetrahydrofuran was added dropwise. After 10 minutes of stirring, the reaction was complete. It was quenched by the addition of 30 mL of a saturated aqueous ammonium chloride solution and extracted with ethyl acetate (3 x 30 mL). The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated and purified by column chromatography (PE:EA = 5:1-1:1) to give the target product as a brown solid (545 mg, 31.1% yield).

[0508] MS ESI: m / z = 344.1, [M+H] + .

[0509] 1 H NMR (400 MHz, CDC13): δ 8.83 (d, 1H), 8.13 (dd, 1H), 7.87 (dd, 1H), 7.48-7.51 (m, 1H), 7.36 (d, 1H), 3.92-3.96 (m, 5H). 2.66-2.69 (m, 2H), 2.49-2.52 (m, 2H), 1.99-2.02 (m, 2H), 1.68-1.71 (m, 2H), 1.52-1.54 (m, 4H).

[0510] Second step: 2-(6-fluoroquinolin-4-yl)-2-hydroxyspiro[3.5]nonan-7-one

[0511]

[0512] The first step product (545 mg, 1.59 mmol) was added to 14 mL of 3 mol / L HCl, stirred at room temperature for 1 h, the reaction was complete. Extracted with ethyl acetate (3 x 5 mL), the aqueous phase was added to an aqueous solution of sodium carbonate, adjusted to pH = 8, extracted with ethyl acetate (3 x 15 mL), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the yellow solid target product (425 mg, yield 89.5%).

[0513] MS ESI: m / z = 300.1, [M+H] + .

[0514] 1 H NMR (400 MHz, CDC13): δ 8.83 (d, 1H), 8.15 (dd, 1H), 7.88 (dd, 1H), 7.48-7.53 (m, 1H), 7.37 (d, 1H), 2.80-2.83 (m, 2H), 2.65-2.68 (m, 2H), 2.42-2.45 (m, 2H), 2.24-2.28 (m, 4H), 1.75-1.78 (m, 2H). One proton that can be exchanged with heavy water was not found by NMR.

[0515] Third step: (±) 2-(6-fluoroquinolin-4-yl)-2-hydroxyspiro[3.5]nonane-7-carbonitrile

[0516]

[0517] The second step product (425 mg, 1.42 mmol), p-methylsulfonylmethyl isocyanide (604 mg, 2.98 mmol) was dissolved in 15 mL of ethylene glycol dimethyl ether, 20 mL of potassium tert-butoxide (699 mg, 6.23 mmol) tert-butanol solution was added at 0°C under nitrogen protection, ethanol (0.25 mL, 4.25 mmol), stirred at room temperature for 3 hours, the reaction was complete. Added 30 mL of ice water, extracted with ethyl acetate (3 x 15 mL), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and column chromatography (PE:EA = 5:1-1:1) to obtain the yellow solid target product (300 mg, yield 71.8%).

[0518] MS ESI: m / z = 311.1, [M+H] + .

[0519] 1H NMR (400 MHz, CDC13): δ 8.81 (d, 1 H), 8.13 (dd, 1 H), 7.85 (dd, 1 H), 7.46-7.51 (m, 1 H), 7.33 (d, 1 H), 2.48-2.69 (m, 5 H), 1.57-1.65 (m, 8 H).

[0520] Fourth step: (±) 2-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-7-carboxylic acid

[0521]

[0522] The third step product (500 mg, 0.97 mmol), red phosphorus (105 mg, 3.40 mmol) were mixed in 4 mL of 55% concentrated hydroiodic acid, stirred at 140 °C under nitrogen protection for 2 hours, the reaction was complete, cooled, filtered to remove red phosphorus, added 2.25 g of sodium carbonate to neutralize the acid, added 1.0 g of sodium thiosulfate to remove elemental iodine, then added 2 mol / L NaOH aqueous solution to adjust to pH = 3, the precipitated solid was filtered to obtain 400 mg of crude product, which was washed with petroleum ether three times, and the solid was filtered to obtain a yellow solid (170 mg, yield 56.3%).

[0523] MS ESI: m / z = 314.1, [M+H] + .

[0524] 1 H NMR (400 MHz, DMSO-d6): δ 12.00 (s, 1 H), 8.81 (d, 1 H), 8.08 (dd, 1 H), 7.74 (dd, 1 H), 7.63-7.68 (m, 1 H), 7.45 (d, 1 H), 4.07-4.11 (m, 1 H), 2.45-2.49 (m, 1 H), 2.30-2.35 (m, 1 H), 2.09-2.18 (m, 2 H), 1.80-1.99 (m, 3 H), 1.64-1.69 (m, 1 H), 1.44-1.60 (m, 3 H), 1.33-1.40 (m, 2 H).

[0525] Fifth step:

[0526] 22A

[0527] N-(4-chlorophenyl)-2-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-7-carboxamide

[0528] 22B

[0529] N-(4-chlorophenyl)-2-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-7-carboxamide

[0530]

[0531] Rac 22: The product of the fourth step (30 mg, 0.096 mmol), triethylamine (0.040 mL, 0.288 mmol), dichloromethane (3 mL) were added to a flask, 2-(7-oxobenzotriazol-1- yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (55 mg, 0.144 mmol) was added, stirred at room temperature for 1 hour, p-chloroaniline (37 mg, 0.288 mmol) was added, stirred at room temperature for 24 hours, the reaction was complete. Concentrated, passed through a column (PE:EA = 5:1-1:1), 30 mg of crude product was obtained, washed with n-hexane three times, filtered to obtain a yellow solid (25 mg, yield 61.1%).

[0532] MS ESI: m / z = 423.1, [M+H] + .

[0533] 1 H NMR (400 MHz, DMSO-d6): δ 9.96 (s, 1H), 8.82 (d, 1H), 8.07-8.10 (m, 1H), 7.76 (d, 1H), 7.63-7.65 (m, 3H), 7.47 (d, 1H), 7.33 (d, 2H), 4.09-4.13 (m, 1H), 2.21-2.33 (m, 3H), 1.78-2.02 (m, 4H), 1.33-1.63 (m, 6H).

[0534] Fifth step isomer A preparation: 20 mg of rac 22 was resolved with chiral column AD-H (n-hexane: isopropanol, 70:30), the retention time of 12 minutes was 22A (4.57 mg, yield 22.9%).

[0535] MS ESI: m / z = 423.1, [M+H] + .

[0536] 1 H NMR (400 MHz, CD3OD): δ 8.76 (d, 1H), 8.07 (dd, 1H), 7.68 (dd, 1H), 7.55-7.61 (m, 3H), 7.50 (d, 1H), 7.28 (d, 2H), 4.11-4.16 (m, 1H), 2.61-2.67 (m, 1H), 2.28-2.45 (m, 3H), 2.03-2.11 (m, 2H), 1.45-1.90 (m, 7H). One proton that can be exchanged by heavy water was not found by NMR.

[0537] Fifth step isomer B preparation: racemic 22 take 20 mg, split by chiral column AD-H (n-hexane: isopropanol, 70:30), the retention time of 20 min is 22B (4.67 mg, yield 23.4%).

[0538] MS ESI: m / z = 423.1, [M+H] + .

[0539] 1 H NMR (400 MHz, CD3OD): δ 8.76 (d, 1H), 8.07 (dd, 1H), 7.68 (dd, 1H), 7.55-7.61 (m, 3H), 7.50 (d, 1H), 7.28 (d, 2H), 4.11-4.15 (m, 1H), 2.61-2.67 (m, 1H), 2.28-2.45 (m, 3H), 2.03-2.11 (m, 2H), 1.45-1.90 (m, 7H). One proton which can be exchanged by heavy water was not found by NMR.

[0540] Resolution of chiral isomers

[0541] Chiral resolution was performed using Agilent 1260 semi-preparative liquid chromatograph (Chiralpak).

[0542] Resolution of the compound of example 15:

[0543] Chiral column: CHIRALPAK AD-H 10*250 mm; flow rate 3.0 mL / min; detection wavelength 254 nm; collect two enantiomers.

[0544] Conditions: take 20 mg of example 22 sample, dissolved in 6 mL of methanol, injection volume 0.5 mL, eluent n-hexane: isopropanol = 70:30 (volume ratio); the peak time of two optical isomers is 12 min and 20 min respectively.

[0545] Example 23

[0546] (±)N-(4-bromophenyl)-2-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-7-carboxamide

[0547]

[0548] According to the preparation method of example 22, the reagent p-chloroaniline of the fifth step is replaced by p-bromoaniline, and the target compound is obtained under the same conditions.

[0549] MS ESI: m / z = 467.0, 469.0, [M+H] + .

[0550] 1 H NMR (400 MHz, CD3OD): δ 8.77 (d, 1 H), 8.07 (dd, 1 H), 7.68 (d, 1 H), 7.62-7.55 (m, 1 H), 7.54-7.47 (m, 3 H), 7.43 (d, 2 H), 4.19-4.08 (m, 1 H), 2.69-2.59 (m, 1 H), 2.45-2.36 (m, 1 H), 2.36-2.25 (m, 2 H), 2.07 (dd, 2 H), 1.93-1.84 (m, 1 H), 1.84-1.66 (m, 3 H), 1.65-1.44 (m, 3 H). One proton which can be exchanged by heavy water was not found by NMR.

[0551] Example 24

[0552] (±) N-(4-fluorophenyl)-2-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-7-carboxamide

[0553]

[0554] Following the procedure of Example 22, the reagent p-chloroaniline in step 5 was replaced by p-fluoroaniline and the same conditions were used to give the target compound.

[0555] MS ESI: m / z = 407.1, [M+H] + .

[0556] 1H NMR (400 MHz, CD3OD): δ 8.76 (d, 1 H), 8.07 (dd, 1 H), 7.68 (dd, 1 H), 7.62-7.47 (m, 4 H), 7.03 (t, 2 H), 4.13 (dt, 1 H), 2.68-2.60 (m, 1 H), 2.46-2.38 (m, 1 H), 2.38-2.25 (m, 2 H), 2.07 (dd, 2 H), 1.94-1.82 (m, 1 H), 1.82-1.64 (m, 3 H), 1.64-1.44 (m, 3 H). One proton which can be exchanged by heavy water was not found by NMR.

[0557] Example 25

[0558] N-(4-fluorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0559]

[0560] First step: (S)-1-phenylethyl 7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxylate

[0561]

[0562] The product of Example 15, eighth step (7.0 g, 22.36 mmol) was dissolved in tetrahydrofuran (110 mL), dicyclohexyl imine (5.54 g, 26.84 mmol) was added followed by 4-dimethylamino pyridine (2.73 g, 22.36 mmol) and stirred for 15 minutes at room temperature. (S)-1-phenylethanol (3.27 g, 26.84 mmol) was added to the reaction and stirred for 24 hours at room temperature. The reaction was complete and worked up. The by-product dicyclohexyl urea was filtered off and the filtrate was concentrated to dryness and purified by column chromatography (Flash: 0-25% ethyl acetate / n-hexane) to give the target product as a colorless liquid (7.0 g, 75% yield).

[0563] MS ESI: m / z = 417.1, [M+H] + .

[0564] Resolution of (S)-1-phenylethyl 7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxylate chiral isomers

[0565] Chiral resolution was performed using Agilent 1260 semi-preparative liquid chromatograph (Chiralpak)

[0566] Chiral column: Chiralpak IC 30*250mm, 5μm; flow rate 1.0 mL / min; detection wavelength 254 nm; two enantiomers were collected.

[0567] Conditions: 1 mg of Example 15 sample 1 was dissolved in 1 mL of methanol, injection volume 0.5 μL, eluent n-hexane: ethanol = 80:20 (volume ratio); the peak time of two optical isomers were 10.89 min and 12.52 min, respectively.

[0568] Isomer A from the first step: 2.98 g was obtained.

[0569] Isomer B from the first step: 2.55 g was obtained.

[0570] Second step: 7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxylic acid

[0571]

[0572] Isomer A from the first step was dissolved in methanol (5.0 mL), replaced with nitrogen once, and palladium hydroxide on carbon (70 mg) was added, followed by the addition of pure water (0.08 mL). The reaction was stirred at room temperature for 16 hours. The reaction was monitored by LCMS and was complete. The palladium catalyst was filtered off using celite and the filtrate was concentrated and purified by column chromatography (Flash: 0-10% methanol / dichloromethane) to give a white solid (340.0 mg, 90.0% yield), [α]D 25 = -31.8448 (c = 0.50, CHCI3).

[0573] MS ESI: m / z = 314.1, [M+H] + .

[0574] Third Step: N-(4-Fluorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2- carboxamide

[0575]

[0576] Take the product in the second step (30 mg, 0.096 mmol), 4-fluoroaniline (24 mg, 0.19 mol), 1-ethyl-(3-dimethylaminopropyl) carbonyldiimide hydrochloride (37 mg, 0.19 mmol) and 1-hydroxybenzotriazole (19 mg, 0.14 mmol), add dichloromethane (5.0 mL), add triethylamine (29 mg, 0.29 mmol) in ice bath under stirring, return to room temperature naturally and stir for 16 hours. Add the reaction solution to water (10 mL), then add ethyl acetate (30 mL), extract with saturated sodium bicarbonate solution (20 mL*2), saturated brine (20 mL*1), and dry over anhydrous sodium sulfate. Filter, evaporate to dryness, and purify by column chromatography (Flash: 0-60% ethyl acetate / n-hexane) to obtain the target product (30 mg, yield 74%)

[0577] MS ESI: m / z = 407.1, [M+H] + .

[0578] 1 H NMR (400 MHz, CDC13): δ 8.80 (d, 1H), 8.13-8.10 (dd, 1H), 7.67-7.64 (dd, 1H), 7.52-7.45 (m, 3H), 7.28-7.27 (m, 1H), 7.04-7.00 (m, 3H), 3.15-3.07 (m, 2H), 2.27-2.20 (m, 3H), 2.11-2.07 (m, 1H), 2.04-1.99 (m, 2H), 1.96-1.93 (m, 1H), 1.87-1.85 (m, 1H), 1.71-1.61 (m, 4H).

[0579] Example 26

[0580] N-(4-Bromophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0581]

[0582] Following the procedure of Example 25, Step 3, using 4-bromoaniline instead of 4-fluoroaniline to afford the title product (35 mg, 86% yield).

[0583] MS ESI: m / z = 487.1, [M+H] + .

[0584] 1 H NMR (400 MHz, CDC13): δ 8.80 (d, 1 H), 8.13-8.10 (dd, 1 H), 7.67-7.64 (dd, 1 H), 7.50-7.44 (m, 5 H), 7.28-7.27 (m, 1 H), 7.04 (s, 1 H), 3.13-3.09 (m, 2 H), 2.23-2.20 (m, 3 H), 2.11-2.07 (m, 1 H), 2.04-1.92 (m, 2 H), 1.96-1.92 (m, 1 H), 1.88-1.85 (m, 1 H), 1.71-1.59 (m, 4 H).

[0585] Example 27

[0586] 7-(6-Fluoroquinolin-4-yl)-N-(pyridin-2-yl)spiro[3.5]nonane-2-carboxamide

[0587]

[0588] Take the carboxylic acid of Example 25, Step 2 (30 mg, 0.096 mmol), 2-aminopyridine (11.6 mg, 0.12 mol), add N-methylimidazole (16.6 mg, 0.20 mmol), add MeCN (2.0 mL), then add N,N,N',N'-tetramethyluronium hexafluorophosphate (32 mg, 0.115 mmol), and allow to stir at room temperature for 16 h. Add the reaction to water (10 mL), then add ethyl acetate (30 mL), and extract with the organic phase with saturated sodium bicarbonate solution (20 mL*2), saturated brine (20 mL*1), and dry over anhydrous sodium sulfate. Filter and evaporate, and purify by column chromatography (Flash: 0-60% ethyl acetate / n-hexane) to afford the title product (35 mg, 86% yield).

[0589] MS ESI: m / z = 390.1, [M+H] + .

[0590] 1H NMR (400 MHz, CDC13): δ 8.80 (d, 1 H), 8.27-8.24 (t, 2H), 8.13-8.10 (dd, 1 H), 7.77 (s, 1 H), 7.73-7.69 (t, 1 H), 7.67-7.64 (dd, 1 H), 7.50-7.44 (td, 1 H), 7.28-7.27 (m, 1 H), 7.05-7.02 (t, 1 H), 3.19-3.12 (m, 2H), 2.27-2.22 (m, 3H), 2.14-2.09 (m, 1 H), 2.06-1.99 (m, 2H), 1.96-1.93 (m, 1 H), 1.88-1.84 (m, 1 H), 1.72-1.60 (m, 4H).

[0591] Example 28

[0592] 7-(6-Fluoroquinolin-4-yl)-N-(3-methyl-4-(l-methyl-lH-pyrazol-3-yl)phenyl)spiro[3.5]nonane-2-carboxamide

[0593]

[0594] Following the procedure of Example 25, Step 3, using 3-fluoro-4-(l-methyl-lH- pyrazol-3-yl)aniline in place of p-fluoroaniline afforded the title product (35 mg, 86% yield).

[0595] MS ESI: m / z = 487.1, [M+H] + .

[0596] 11 H NMR (400 MHz, CDC13): δ 8.80 (d, 1 H), 8.27-8.24 (t, 2H), 8.13-8.10 (dd, 1 H), 7.77 (s, 1 H), 7.73-7.69 (t, 1 H), 7.67-7.64 (dd, 1 H), 7.50-7.44 (td, 1 H), 7.28-7.27 (m, 1 H), 7.05-7.02 (t, 1 H), 3.19-3.12 (m, 2H), 2.27-2.22 (m, 3H), 2.14-2.09 (m, 1 H), 2.06-1.99 (m, 2H), 1.96-1.93 (m, 1 H), 1.88-1.84 (m, 1 H), 1.72-1.60 (m, 4H).

[0597] Example 29

[0598] N-(2-Chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0599]

[0600] Following the procedure for Example 25, substituting 4-chloro-2- fluorobenzenamine for 4-fluorobenzenamine in the third step, and using the same conditions to obtain the target compound.

[0601] MS ESI: 441.1 [M+H] + .

[0602] 1 H NMR (400 MHz, CDC13 ) δ 8.81 (d, 1H), 8.44 (d, 1H), 8.12 (dd, 1H), 7.66 (dd, 1H), 7.61 (s, 1H), 7.47 (ddd, 1H), 7.37 (dd, 1H), 7.33 - 7.26 (m, 2H), 7.04 (td, 1H), 3.21 (p, 1H), 3.16 - 3.07 (m, 1H), 2.33 - 2.19 (m, 3H), 2.18 - 2.10 (m, 1H), 2.09 - 2.02 (m, 1H), 2.02 - 1.90 (m, 2H), 1.90 - 1.81 (m, 1H), 1.74 - 1.66 (m, 1H), 1.65 - 1.55 (m, 3H).

[0603] Example 30

[0604] N-(4-chloro-2-fluorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2- carboxamide

[0605]

[0606] Following the procedure for Example 25, substituting 4-chloro-2- fluorobenzenamine for 4-fluorobenzenamine in the third step, and using the same conditions to obtain the target compound.

[0607] MS ESI: 441.1 [M+H] + .

[0608] 1 H NMR (400 MHz, CDC13 ) δ 8.81 (d, 1H), 8.44 (d, 1H), 8.12 (dd, 1H), 7.66 (dd, 1H), 7.61 (s, 1H), 7.47 (ddd, 1H), 7.37 (dd, 1H), 7.33 - 7.26 (m, 2H), 7.04 (td, 1H), 3.21 (p, 1H), 3.16 - 3.07 (m, 1H), 2.33 - 2.19 (m, 3H), 2.18 - 2.10 (m, 1H), 2.09 - 2.02 (m, 1H), 2.02 - 1.90 (m, 2H), 1.90 - 1.81 (m, 1H), 1.74 - 1.66 (m, 1H), 1.65 - 1.55 (m, 3H).

[0609] Example 31

[0610] N-(3-fluorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0611]

[0612] Following the procedure of Example 25, the reagent 4-fluoroaniline in the third step was replaced with 3-fluoroaniline and the same conditions were used to obtain the target compound.

[0613] MS ESI: m / z = 407.1, [M+H] + .

[0614] 1 H NMR (400 MHz, CDC13): δ 8.81 (d, 1H), 8.10-8.14 (m, 1H), 7.65 (dd, 1H), 7.45-7.55 (m, 2H), 7.27-7.29 (m, 2H), 7.11-7.16 (m, 2H), 6.80-6.83 (m, 1H), 3.10-3.14 (m, 2H), 2.20-2.27 (m, 3H), 2.07-2.12 (m, 1H), 1.85-2.01 (m, 4H), 1.56-1.71 (m, 4H).

[0615] Example 32

[0616] N-(3-bromophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0617]

[0618] Following the procedure of Example 25, the reagent 4-fluoroaniline in the third step was replaced with 3-bromoaniline and the same conditions were used to obtain the target compound.

[0619] MS ESI: m / z = 467.0, [M+H] + .

[0620] 1H NMR (400 MHz, CDC13): δ 8.81 (d, 1 H), 8.12-8.15 (m, 1 H), 7.84 (s, 1 H), 7.65 (dd, 1 H), 7.42-7.50 (m, 2 H), 7.27-7.29 (m, 1 H), 7.16-7.23 (m, 2 H), 7.03 (s, 1 H), 3.10-3.13 (m, 2 H), 2.20-2.27 (m, 3 H), 2.07-2.12 (m, 1 H), 1.85-2.07 (m, 4 H), 1.56-1.71 (m, 4 H).

[0621] Example 33

[0622] 7-(6-Fluoroquinolin-4-yl)-N-(pyridin-3-yl)spiro[3.5]nonane-2-carboxamide

[0623]

[0624] Following the procedure of Example 25, the reagent 4-fluoroaniline in the third step was replaced with 3-fluoro-4-chloroaniline and the same conditions were used to obtain the target compound.

[0625] MS ESI: m / z = 441.1, [M+H] + .

[0626] 1 H NMR (400 MHz, CDC13): δ 8.81 (d, 1 H), 8.59 (s, 1 H), 8.36 (s, 1 H), 8.27 (d, 1 H), 8.12-8.16 (dd, 1 H), 7.66 (dd, 1 H), 7.46-7.51 (m, 1 H), 7.29-7.32 (m, 2 H), 7.20 (s, 1 H), 3.10-3.19 (m, 2 H), 2.22-2.29 (m, 3 H), 2.10-2.15 (m, 1 H), 1.86-2.06 (m, 4 H), 1.56-1.71 (m, 4 H).

[0627] Example 34

[0628] N-(3-Fluoro-4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0629]

[0630] Following the procedure of Example 25, the reagent 4-fluoroaniline in the third step was replaced with 3-fluoro-4-chloroaniline and the same conditions were used to obtain the target compound.

[0631] MS ESI: m / z = 441.1, [M+H]+ .

[0632] Example 35

[0633] 7-(6-Fluoroquinolin-4-yl)-N-(4-trifluoromethyl)spiro[3.5]nonane-2-carboxamide

[0634]

[0635] Following the procedure of Example 25, the reagent 4-fluoroaniline in the third step was replaced with 4-trifluoromethylaniline and the same conditions were used to obtain the target compound.

[0636] MS ESI: m / z = 457.0, [M+H] + .

[0637] 1H NMR (400 MHz, CDC13) δ 8.81 (d, 1H), 8.13 (dd, 1H), 7.64-7.68 (m, 3H), 7.59 (s, 1H), 7.57 (s, 1H), 7.48 (ddd, 1H), 7.28 (d, 1H), 7.22 (s, 1H), 3.09-3.19 (m, 2H), 2.22-2.28 (m, 3H), 2.11 (t, 1H), 1.99-2.06 (m, 2H), 1.92-1.96 (m, 1H), 1.85-1.88 (m, 1H), 1.68-1.72 (m, 1H), 1.63-1.66 (m, 1H), 1.58-1.62 (m, 2H).

[0638] Example 36

[0639] N-(3-Chlorophenyl)-7-(6-fluoroquinolin-4-yl)-spiro[3.5]nonane-2-carboxamide

[0640]

[0641] Following the procedure of Example 25, the reagent 4-fluoroaniline in the third step was replaced with 3-chloroaniline and the same conditions were used to obtain the target compound.

[0642] MS ESI: m / z = 422.9, [M+H] + .

[0643] Example 37

[0644] N-(3-Fluoro-4-bromophenyl)-7-(6-fluoroquinolin-4-yl)-spiro[3.5]nonane-2-carboxamide

[0645]

[0646] The reagent 4-fluoroaniline in the third step was replaced with 3-chloro-4- fluoronaniline according to the procedure in Example 25 to give the target compound.

[0647] MS ESI: m / z = 485.4, [M+H] + .

[0648] Example 38

[0649] N-(3-chloro-4-fluoro)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0650]

[0651] The reagent 4-fluoroaniline in the third step was replaced with 3-chloro-4- fluoronaniline according to the procedure in Example 25 to give the target compound.

[0652] MS ESI: m / z = 441.1, [M+1] + .

[0653] 1 H NMR (400 MHz, CDC13) δ 8.81 (d, 1H), 8.11 (dd, 1H), 7.76 (dd, 1H), 7.65 (dd, 1H), 7.51 - 7.43 (m, 1H), 7.36 - 7.30 (m, 1H), 7.28 (s, 1H), 7.09 (t, 1H), 7.02 (s, 1H), 3.10 (p, 2H), 2.26 - 2.18 (m, 3H), 2.14 - 2.06 (m, 1H), 2.05 - 1.97 (m, 2H), 1.96 - 1.90 (m, 1H), 1.89 - 1.83 (m, 1H), 1.74 - 1.57 (m, 4H).

[0654] Example 39

[0655] 7-(6-fluoroquinolin-4-yl)-N-(3-(trifluoromethyl)phenyl)spiro[3.5]nonane-2-carboxamide

[0656]

[0657] The reagent 4-fluoroaniline in the third step was replaced with 3-chloro-4- fluoronaniline according to the procedure in Example 25 to give the target compound. MS ESI: m / z = 441.1, [M+1] + .

[0658] 1H NMR (400 MHz, CDC13) δ 8.81 (d, 1 H), 8.11 (dd, 1 H), 7.88 (s, 1 H), 7.72 (d, 1 H), 7.65 (dd, 1 H), 7.50-7.40 (m, 2 H), 7.36 (d, 1 H), 7.25 (s, 1 H), 7.14 (s, 1 H), 3.13 (p, 2 H), 2.29-2.19 (m, 3 H), 2.15-2.07 (m, 1 H), 2.06-1.98 (m, 2 H), 1.97-1.90 (m, 1 H), 1.89-1.83 (m, 1 H), 1.73-1.57 (m, 4 H).

[0659] Example 40

[0660] N-(3,4-difluorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0661]

[0662] Following the procedure of Example 25, the reagent of step 3, 4-fluoroaniline, was replaced with 3,4-difluoroaniline and the same conditions were used to obtain the target compound.

[0663] MS ESI: m / z = 425.1, [M+H] + .

[0664] 1 H NMR (400 MHz, CDC13): δ 8.81 (d, 1 H), 8.12 (dd, 1 H), 7.70-7.62 (m, 2 H), 7.50-7.44 (m, 1 H), 7.28 (s, 1 H), 7.08 (d, 1 H), 7.01 (s, 1 H), 3.16-3.05 (m, 2 H), 2.27-2.19 (m, 3 H), 2.13-2.06 (m, 1 H), 2.06-1.98 (m, 2 H), 1.97-1.91 (m, 1 H), 1.89-1.83 (m, 1 H), 1.73-1.64 (m, 2 H), 1.64-1.57 (m, 2 H). One proton that can be exchanged with heavy water was not found by NMR.

[0665] Example 41

[0666] 7-(6-fluoroquinolin-4-yl)-N-(2-(trifluoromethyl)phenyl)spiro[3.5]nonane-2-carboxamide

[0667]

[0668] The target compound was obtained according to the procedure of Example 27, substituting reagent 2-aminopyridine with 2-amino-5-chloropyridine. MS ESI: m / z = 424.1, [M+H] + .

[0669] 1 H NMR (400 MHz, CDC13) δ 8.81 (d, 1H), 8.27 (d, 1H), 8.12 (dd, 1H), 7.65 (dd, 1H), 7.61 (d, 1H), 7.57 (t, 1H), 7.47 (ddd, 1H), 7.39 (s, 1H), 7.28 (d, 1H), 7.23 (t, 1H), 3.23-3.15 (m, 1H), 3.15-3.08 (m, 1H), 2.31-2.23 (m, 1H), 2.23-2.18 (m, 1H), 2.18-2.11 (m, 1H), 2.09-2.03 (m, 1H), 1.99-1.91 (m, 2H), 1.89-1.83 (m, 1H), 1.73-1.64 (m, 2H), 1.64-1.59 (m, 2H), 1.58-1.52 (m, 1H).

[0670] Example 42

[0671] N-(5-chloropyridin-2-yl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0672]

[0673] The target compound was obtained according to the procedure of Example 27, substituting reagent 2-aminopyridine with 2-amino-5-chloropyridine. MS ESI: m / z = 424.1, [M+H]

[0674] MS ESI: m / z = 424.1, [M+H] + .

[0675] 1 H NMR (400 MHz, CDC13) δ 8.81 (d, 1H), 8.27 (d, 1H), 8.12 (dd, 1H), 7.65 (dd, 1H), 7.61 (d, 1H), 7.57 (t, 1H), 7.47 (ddd, 1H), 7.39 (s, 1H), 7.28 (d, 1H), 7.23 (t, 1H), 3.23-3.15 (m, 1H), 3.15-3.08 (m, 1H), 2.31-2.23 (m, 1H), 2.23-2.18 (m, 1H), 2.18-2.11 (m, 1H), 2.09-2.03 (m, 1H), 1.99-1.91 (m, 2H), 1.89-1.83 (m, 1H), 1.73-1.64 (m, 2H), 1.64-1.59 (m, 2H), 1.58-1.52 (m, 1H).

[0676] Example 43

[0677] 7-(6-Fluoroquinolin-4-yl)-N-(1,1 '-biphenyl-4-yl)spiro[3,5]nonane-2- carboxamide

[0678]

[0679] Following the procedure of Example 25, the reagent p-chloroaniline in the third step was replaced with p-biphenylamine and the same conditions were used to obtain the target compound.

[0680] MS ESI: M / Z = 465.1, [M+1] + .

[0681] 1 H NMR (400 MHz, CDC13): δ 8.81 (d, 1 H), 8.12 (dd, 1 H), 7.68-7.61 (m, 3H), 7.57 (dd, 4H), 7.50-7.46 (m, 1 H), 7.43 (t, 2H), 7.33 (t, 1 H), 7.28 (d, 1 H), 7.12 (s, 1 H), 3.14 (m, 2H), 2.30-2.19 (m, 3H), 2.11 (m, 1 H), 2.03 (m, 2H), 1.95 (dd, 1 H), 1.87 (d, 1 H), 1.74-1.59 (m, 4H).

[0682] Example 44

[0683] 7-(6-Fluoroquinolin-4-yl)-N-(1,1 '-biphenyl-4-yl)spiro[3,5]nonane-2- carboxamide

[0684]

[0685] Following the procedure of Example 25, the reagent p-chloroaniline in the third step was replaced with 3-biphenylamine and the same conditions were used to obtain the target compound.

[0686] MS ESI: M / Z = 465.1, [M+1] + .

[0687] 1 ​H NMR (400 MHz, CDC13) δ 8.81 (d, 1 H), 8.12 (dd, 1 H), 7.85 (s, 1 H), 7.66 (dd, 1 H), 7.62-7.58 (m, 2 H), 7.52-7.32 (m, 7 H), 7.28 (d, 1 H), 7.12 (s, 1 H), 3.20-3.08 (m, 2 H), 2.31-2.21 (m, 3 H), 2.09 (m, 2 H), 1.97-1.83 (m, 3 H), 1.75-1.59 (m, 4 H).

[0688] Example 45

[0689] N-(3-chloro-4-(trifluoromethyl)phenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2- carboxamide

[0690]

[0691] Following the procedure of Example 27, reagent 2-aminopyridine was replaced with 3-chloro-4-trifluoromethyl-aniline to give the target compound 37.9 mg, yield: 81.8%.

[0692] MS ESI: m / z = 491.0, [M+H] + .

[0693] 1 H NMR (400 MHz, CDC13) δ 8.81 (d, 1 H), 8.12 (dd, 1 H), 7.85 (s, 1 H), 7.66 (dd, 1 H), 7.62-7.58 (m, 2 H), 7.52-7.32 (m, 7 H), 7.28 (d, 1 H), 7.12 (s, 1 H), 3.20-3.08 (m, 2 H), 2.31-2.21 (m, 3 H), 2.09 (m, 2 H), 1.97-1.83 (m, 3 H), 1.75-1.59 (m, 4 H).

[0694] Example 46

[0695] N-(2-fluorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0696]

[0697] Following the procedure of Example 25, reagent 4-fluoroaniline in Step 3 was replaced with 2-fluoroaniline to give the target compound under the same conditions.

[0698] 1H NMR (400 MHz, CDC13): δ 8.81 (d, 1 H), 8.39 (t, 1 H), 8.12 (dd, 1 H), 7.65 (d, 1 H), 7.47 (t, 1 H), 7.32-7.26 (m, 2 H), 7.17-6.99 (m, 3 H), 3.23-3.06 (m, 2 H), 2.25 (dd, 3 H), 2.15-1.82 (m, 6 H), 1.67 (dd, 3 H).

[0699] MS ESI: m / z = 407.1, [M+H] + .

[0700] Example 47

[0701] N-(2-bromophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide

[0702]

[0703] Following the preparation method of Example 27, reagent 4-fluoroaniline was replaced with 2-bromoaniline and the same conditions were used to obtain the target compound. 1 H NMR (400 MHz, CDC13): δ 8.81 (d, 1 H), 8.42 (d, 1 H), 8.12 (dd, 1 H), 7.70-7.58 (m, 2 H), 7.54 (dd, 1 H), 7.50-7.42 (m, 1 H), 7.37-7.30 (m, 1 H), 7.28 (d, 1 H), 6.97 (dd, 1 H), 3.28-3.06 (m, 2 H), 2.33-2.21 (m, 3 H), 2.02 (m, 6 H), 1.74-1.62 (m, 3 H).

[0704] MS ESI: m / z = 467.0, [M+H] + .

[0705] Example 48 7-(6-fluoroquinolin-4-yl)-N-(pyridin-4-yl)spiro[3.5]nonane-2-carboxamide

[0706]

[0707] Following the preparation method of Example 27, reagent 4-fluoroaniline was replaced with 4-aminopyridine and the same conditions were used to obtain the target compound. 1H NMR (400 MHz, CDC13) δ 8.81 (d, 1 H), 8.51 (s, 2 H), 8.12 (dd, 1 H), 7.65 (dd, 1 H), 7.50 (d, 1 H), 7.49 (s, 1 H), 7.48-7.44 (m, 1 H), 7.35 (s, 1 H), 7.27 (d, 1 H), 3.16 (dd, 2 H), 2.29-2.18 (m, 3 H), 2.15-1.82 (m, 6 H), 1.64 (s, 3 H). MS ESI: m / z = 390.1, [M+H] + .

[0708] Example 49 7-(6-Fluoroquinolin-4-yl)-N-(4-cyanophenyl)spiro[3,5]nonane-2- carboxamide

[0709]

[0710] The reagents, the reagent 4-fluoroaniline was replaced with p-cyanophenylamine, using the same conditions to give the target compound. MS ESI: M / Z = 414.1, [M+1] + .

[0711] 1 H NMR (400 MHz, CDC13): δ 8.81 (d, 1 H), 8.12 (dd, 1 H), 7.72-7.59 (m, 4 H), 7.51-7.43 (m, 1 H), 7.27 (d, 2 H), 7.24 (s, 1 H), 3.14 (dd, 2 H), 2.29-2.19 (m, 3 H), 2.16-2.07 (m, 1 H), 2.04 (s, 1 H), 1.95 (dd, 2 H), 1.87 (d, 1 H), 1.74-1.63 (m, 4 H).

[0712] Example 50 7-(6-Fluoroquinolin-4-yl)-N-phenylspiro[3.5]nonane-2-carboxamide

[0713]

[0714] Following the procedure of Example 25, Step 3, aniline was used instead of p-fluoroaniline to give the target product (30 mg, yield 81%). MS ESI: m / z = 389.1, [M+H] + . 1H NMR (400 MHz, CDC13): δ 8.80 (d, 1 H), 8.13-8.10 (dd, 1 H), 7.67-7.64 (dd, 1 H), 7.54 (d, 2 H), 7.49-7.44 (td, 1 H), 7.35-7.31 (t, 2 H), 7.28-7.27 (m, 1 H), 7.13-7.09 (t, 1 H), 7.04 (s, 1 H), 3.17-3.08 (m, 2 H), 2.28-2.21 (m, 3 H), 2.12-2.08 (m, 1 H), 2.05-2.00 (m, 2 H), 1.96-1.92 (m, 1 H), 1.87-1.84 (m, 1 H), 1.71-1.59 (m, 4 H).

[0715] Example 51 N-(4-Fluorophenyl)-2-(6-fluoroquinolin-4-yl)-7-azaspiro[3.5]nonane-7- carboxamide

[0716]

[0717] First Step: 2-(6-Fluoroquinolin-4-yl)-2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester

[0718]

[0719] To a solution of 4-bromo-6-fluoroquinoline (500.0 mg, 2.21 mmol) in dry tetrahydrofuran (44 mL) under argon was added tert-butyllithium solution (2.76 mL, 1.6 M, 4.42 mmol) slowly dropwise at -78 °C. After the addition was complete, the reaction was stirred for 3 minutes, and then a solution of 2-oxo-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (528.8 mg, 2.21 mmol) in tetrahydrofuran (4.0 mL) was added. After the addition was complete, the reaction was stirred at this temperature for 30 minutes. The reaction was quenched by the addition of acetic acid (133 mg), and the reaction was concentrated and purified by column chromatography (Flash: 0-100% ethyl acetate in hexanes) to give a light yellow solid (210 mg, 25% yield). MS ESI: m / z = 387.1, [M+H] + .

[0720] 1H NMR (400 MHz, CDC13): δ 8.86 (d, 1 H), 8.16-8.13 (dd, 1 H), 7.87-7.85 (dd, 1 H), 7.52-7.49 (m, 1 H), 7.49 (d, 1 H), 3.75-3.70 (m, 2 H), 3.45-3.43 (m, 2 H), 3.28-3.26 (m, 2 H), 2.70-2.51 (m, 4 H), 2.24-2.20 (m, 1 H), 1.91-1.88 (m, 2 H), 1.45 (s, 9 H).

[0721] Second Step: 6-Fluoro-4-(7-azaspiro[3.5]nonan-2-yl)quinoline

[0722]

[0723] The product of the first step (210 mg, 0.544 mmol) was placed in a reaction flask, concentrated hydroiodic acid (mass fraction: 55%) (2.0 mL) was added, followed by red phosphorus (84.0 mg, 2.72 mmol), and heated to a reaction temperature of 140 °C quickly, and reacted for 5 hours. TLC and LCMS detection showed that the reaction was complete. 1 mol / L aqueous sodium hydroxide solution was added to adjust the pH to about 10, and then aqueous sodium sulfite solution (20.0 mL) was added, stirred for 5 minutes, and no color change was observed with starch iodide paper. Dichloromethane (30.0 mL) was added for extraction, and the extraction was repeated twice. The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product (140 mg, yield 95%). MS ESI: m / z = 271.1, [M+H] + .

[0724] 1 H NMR (400 MHz, CDC13): δ 8.82 (d, 1 H), 8.13-8.09 (m, 1 H), 7.48-7.44 (m, 2 H), 7.28-7.27 (m, 1 H), 4.03-3.98 (m, 1 H), 3.66 (s, 1 H), 3.01-2.98 (m, 2 H), 2.84-2.81 (m, 2 H), 2.54-2.49 (m, 2 H), 2.10-2.05 (m, 2 H), 1.94-1.91 (m, 2 H), 1.63-1.60 (m, 2 H).

[0725] Third Step: N-(4-Fluorophenyl)-2-(6-fluoroquinolin-4-yl)-7-azaspiro[3.5]nonane-7- carboxamide

[0726]

[0727] The product of the second step (30.0 mg, 0.11 mmol) was placed in a reaction vial, N,N-diisopropylethylamine (21.5 mg, 30 uL) was added, after stirring for 2 minutes in an ice bath, 4-fluorophenyl isocyanate (16.7 mg, 0.12 mmol) was added, and the reaction was allowed to proceed at room temperature for 3 hours. TLC, LCMS monitoring. The solvent was evaporated, the sample was swirled, and column chromatography (Flash: 0-10% methanol / dichloromethane) was performed to obtain the white target product.

[0728] MS ESI: m / z = 408.1, [M+H] + .

[0729] Example 52

[0730] N-(4-chlorophenyl)-2-(6-fluoroquinolin-4-yl)-7-azaspiro[3.5]nonane-7-carboxamide

[0731]

[0732] Following the procedure of Example 51, the reagent 4-fluorophenyl isocyanate in the third step was replaced with 4-chlorophenyl isocyanate, and the target compound was obtained under the same conditions.

[0733] MS ESI: m / z = 424.1, [M+H] + .

[0734] Example 53

[0735] N-(4-bromophenyl)-2-(6-fluoroquinolin-4-yl)-7-azaspiro[3.5]nonane-7-carboxamide

[0736]

[0737] Following the procedure of Example 51, the reagent 4-fluorophenyl isocyanate in the third step was replaced with 4-bromophenyl isocyanate, and the target compound was obtained under the same conditions.

[0738] MS ESI: m / z = 468.1, 470.1 [M+H] + .

[0739] Example 54

[0740] N-(4-cyanophenyl)-2-(6-fluoroquinolin-4-yl)-7-azaspiro[3.5]nonane-7-carboxamide

[0741]

[0742] Following the procedure of Example 51, the reagent 4-fluorophenyl isocyanate in the third step was replaced with 4-cyanophenyl isocyanate, and the target compound was obtained under the same conditions.

[0743] MS ESI: m / z = 415.1, [M+H] + .

[0744] Example 55

[0745] N-(4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)-2-azaspiro[3.5]nonane-2- carboxamide

[0746]

[0747] First step: (6-fluoroquinolin-4-yl)boronic acid

[0748]

[0749] Into a three-necked flask was placed 4-bromo-6-fluoroquinoline (1.00 g, 4.42 mmol), triisopropyl borate (3.06 mL, 13.25 mmol) and 20 mL of dry tetrahydrofuran, replaced with nitrogen 3 times, cooled to -78 °C with dry ice acetone bath, added 1.6 mol / L tert-butyllithium in n-heptane solution (5.53 mL, 8.84 mmol) at -78 °C under nitrogen protection, stirred at -78 °C under nitrogen protection for 0.5 h, the reaction was complete, added saturated ammonium chloride solution (20 mL) to quench, extracted with 20 mL of ethyl acetate for 3 times, the organic phase was washed with saturated brine (20 mL), dried over sodium sulfate, filtered, the filtrate was concentrated to give the crude product 1.00 g, the solid was washed with petroleum ether: ethyl acetate = 1: 1 solution to give the target yellow solid product (400 mg, yield 47.4%). MS ESI: m / z = 192.1, [M+H] + .

[0750] Second step: tert-butyl 7-(((trifluoromethyl)sulfonyl)oxy)-2-azaspiro[3.5]non-6- ene-2-carboxylate

[0751]

[0752] Into a 50 mL two-necked flask, 7-oxo-2-azaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (306 mg, 1.28 mmol), N-phenyl bis(trifluoromethanesulfonimide) (507 mg, 1.42 mmol) were added, and super dry tetrahydrofuran (20 mL) was added under nitrogen protection. The flask was cooled to -78 °C in a dry ice acetone bath, and then 2 mol / L sodium bis(trimethylsilyl)amide in tetrahydrofuran (0.77 mL, 1.54 mmol) was slowly added dropwise into the flask. The addition was completed in 5 minutes. After 2 hours of reaction, the reaction was monitored by TLC and was complete. Saturated ammonium chloride solution (2 mL) was added to quench the reaction. The reaction solution was dried by rotary evaporation, and ethyl acetate (30 mL) was added. The ethyl acetate phase was washed with 5% sodium hydroxide solution (30 mL) three times, and then dried. The crude product (724 mg) was obtained by rotary evaporation and was directly used in the next step. 1 H NMR (400 MHz, CDC13): δ 5.67-5.73 (m, 1H), 3.70 (d, 2H), 3.64 (d, 2H), 2.36-2.46 (m, 4H), 1.93-1.98 (m, 2H), 1.44 (s, 9H).

[0753] Step 3: 7-(6-fluoroquinolin-4-yl)-2-azaspiro[3,5]non-6-ene-2-carboxylic acid tert-butyl ester

[0754]

[0755] Into a three-necked flask, the product of the first step (400 mg, 2.09 mmol), the crude product of the second step (724 mg), and potassium carbonate (354 mg, 2.56 mmol) were added. Then 20 mL of dioxane and 2 mL of water were added, and the flask was purged with nitrogen three times. Tetraphenylphosphonium palladium (75 mg, 0.065 mmol) was added, and the flask was purged with nitrogen three times. The reaction solution was heated to 100 °C, and stirred at 100 °C under nitrogen protection for 4 h. The reaction was complete. The reaction solution was concentrated, and the product was purified by column chromatography (petroleum ether: ethyl acetate = 5:1-2:1) to obtain the yellow target product (330 mg, 72.8%). MS ESI: m / z = 369.1, [M+H] + .

[0756] Step 4: 7-(6-fluoroquinolin-4-yl)-2-azaspiro[3,5]nonane-2-carboxylic acid tert-butyl ester

[0757]

[0758] The third step product (330 mg, 0.90 mmol), methanol (20 mL), was added to a flask, 10% palladium carbon (33 mg) was added, stirred under normal temperature hydrogen environment for 16 h, the reaction was incomplete. 10% palladium carbon (33 mg) was added, stirred under normal temperature hydrogen environment for 16 h, the reaction was incomplete, 10% palladium carbon (264 mg) was added, stirred under normal temperature hydrogen environment for 12 h, the reaction was incomplete, the reaction liquid was filtered, the filtrate was concentrated, and column chromatography (petroleum ether: ethyl acetate = 10:1-3:2) was performed to obtain the white solid target product (120 mg, yield 36.4%).

[0759] MS ESI: m / z = 371.1, [M+H] + .

[0760] Step 5: 6-Fluoro-4-(2-aza-spiro[3.5]non-7-yl)quinoline trifluoroacetate

[0761]

[0762] The fourth step product (120 mg, 0.32 mmol), 5 mL of dichloromethane, was added to a flask, 1 mL of trifluoroacetic acid was added, stirred at room temperature for 0.5 h, the reaction was complete, and concentration was performed to obtain the yellow target product.

[0763] MS ESI: m / z = 271.1, [M+H] + .

[0764] Step 6: N-(4-chlorophenyl)-7-(6-fluoroquinolin-4-yl)-2-azaspiro[3.5]nonane-2- carboxamide

[0765]

[0766] The fifth step product (40 mg, 0.08 mmol), triethylamine (0.039 mL, 0.28 mmol), 2 mL of dichloromethane, was added to a flask, p-chlorophenyl isocyanate (15 mg, 0.096 mmol) was added, stirred at room temperature for 1 h, the reaction was complete, concentrated, and column chromatography was performed to obtain the target product. MS ESI: m / z = 424.1, [M+H] + .

[0767] Example 56

[0768] N-(4-bromophenyl)-7-(6-fluoroquinolin-4-yl)-2-azaspiro[3.5]nonane-2-carboxamide

[0769]

[0770] Following the procedure of Example 55, the reagent p-chlorophenyl isocyanate in Step 6 was replaced with 4-bromophenyl isocyanate, and the target compound was obtained under the same conditions.

[0771] MS ESI: m / z = 468.0, 470.0, [M+H] + .

[0772] Example 57

[0773] N-(4-cyanophenyl)-7-(6-fluoroquinolin-4-yl)-2-azaspiro[3.5]nonane-2-carboxamide

[0774]

[0775] Following the procedure of Example 55, the reagent p-chlorophenyl isocyanate in Step 6 was replaced with 4-cyanophenyl isocyanate, and the target compound was obtained under the same conditions.

[0776] MS ESI: m / z = 415.1, [M+H] + .

[0777] Example 58

[0778] N-(4-cyanophenyl)-7-(6-fluoroquinolin-4-yl)-2-azaspiro[3.5]nonane-2-carboxamide

[0779]

[0780] Following the procedure of Example 55, the reagent p-chlorophenyl isocyanate in Step 6 was replaced with 4-fluorophenyl isocyanate, and the target compound was obtained under the same conditions. MS ESI: m / z = 408.1, [M+H] + .

[0781] Example 59 Activity test

[0782] (1) Induction expression and purification method of IDO protein

[0783] First, the IDO gene was amplified by PCR, and the amplified PCR product was recovered. Then, the pET28a plasmid (purchased from Shanghai Baoman Biotechnology Co., Ltd.) and the IDO gel recovery product were digested with EcoR I and Xho I restriction enzymes (37°C, enzyme digestion for 2h), and the gel was run, recovered, and T4 ligase was linked overnight. The product was added to the DH5a competent cells, placed on ice for 30 min, heat shocked at 42°C for 90s, shaken and plated, and single colonies were picked and identified by PCR and sequencing. All correct, that is, the pET28a-IDO plasmid was successfully constructed.

[0784] The constructed BL21 containing pET28a-IDO plasmid was shaken at 37°C until OD 600 was 0.6-0.8, hemin was added to a final concentration of 40 μM, IPTG (isopropyl-β-D- thiogalactoside) was added to a final concentration of 0.5 mM, and induction was performed at 16°C for 20 h; after induction, the bacterial cells were collected by centrifugation at 4°C and 6000 rpm, and the collected bacterial cells were washed once with 50 mM PBS (pH 7.5) and then collected by centrifugation.

[0785] The collected bacterial cells were resuspended with buffer (50 mM PBS pH 7.5) and an appropriate amount of 100×PMSF was added, the cells were broken by a cell disrupter at 1400 bar and 4°C for three times, the lysed bacteria were centrifuged at 18000 x g for 45 min, the precipitate was discarded, the supernatant was retained, and the supernatant was filtered through a 0.45 μm membrane at 4°C; the nickel column was equilibrated with 3 column volumes of lysis buffer (50 mM PBS pH 7.5), then the lysis supernatant was loaded onto the nickel column, after loading, the column was washed with 4 column volumes of washing buffer (50 mM PBS pH 7.5, 50 mM imidazole), and finally the protein was eluted with elution buffer (50 mM PBS pH 7.5, 250 mM imidazole); the eluted protein solution was dialyzed for 6 h in 50 mM PBS pH 7.5, after dialysis, the protein sample was concentrated, aliquoted, quickly frozen in liquid nitrogen, and stored at -80°C for standby use.

[0786] (2) Test method for IDO enzyme inhibitory activity

[0787] First, the compound was diluted by 3-fold gradient, 1 μL of each concentration was added to a 96-well plate; 50 μL of prepared IDO enzyme solution PBS (pH 7.5) was added to a final concentration of 50 mM; 25 μL of substrate 1 (methylene blue, final concentration 3.5 μM; catalase, final concentration 0.2 μg / μL; PBS (pH 7.5), final concentration 50 mM) mixed solution was added, and 25 μL of substrate 2 (D-Trp, final concentration 1.5 mM; sodium ascorbate, final concentration 20 mM; PBS (pH 7.5), final concentration 50 mM) mixed solution was added to start the reaction. Finally, OD 321 nm reading was performed for 40 min.

[0788] (3) Test method for cell activity

[0789] Hela cells (80 μL) were inoculated in a 96-well plate, and the inoculation amount was 5 x 10 3The compounds were diluted and 1 μL was added to the 96-well plates. Medium containing human interferon gamma (final concentration 50 ng / mL) was added to the 96-well plates to a final volume of 200 μL. After 48 hours incubation, 80 μL of supernatant was transferred to a new 3894-well plate. 10 μL of 6.1 N trichloroacetic acid was added to each well and mixed, and incubated at 50°C for 30 minutes. The IDO enzyme catalyzes the conversion of N-formylkynurenine to kynurenine. The reaction mixture was centrifuged at 2500 rpm for 10 minutes, and 70 μL of supernatant was transferred to a new 96-well plate and mixed with 100 μL of 2% (w / v) dimethylaminobenzaldehyde acetic acid solution. After 5-10 minutes, the values were measured at 480 nm using a SPECTRAmax microplate reader.

[0790] The results of the tests of the IDO enzyme inhibitory activity and the cytostatic activity of the compounds are shown in Table 1.

[0791] Table 1. Results of tests of IDO enzyme and cytostatic activity

[0792]

[0793]

[0794] The above results show that the compounds of the present application have inhibitory activity against the IDO enzyme and cells.

[0795] All documents referred to in this application are incorporated herein by reference as if each were individually incorporated by reference. In addition, it is to be understood that the application can be carried out by specifically different embodiments and that embodiments can be practiced without the specific details (e.g., specific process parameters, washing procedures, etc.) that are set forth in the preceding description. Therefore, it will be appreciated that departures from these specific details can still be made and fall within the scope of the application.

Claims

1. A compound, its stereoisomer, tautomer, or pharmaceutically acceptable salt, characterized in that: The compound is N -(4-Fluorophenyl)-7-(6-fluoroquinolin-4-yl)spiro[3.5]nonane-2-carboxamide.

2. The compound according to claim 1, its stereoisomer, tautomer, or pharmaceutically acceptable salt thereof, wherein: The pharmaceutically acceptable salt is selected from the group consisting of hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, trifluoromethanesulfonate, benzenesulfonate, p-toluenesulfonate (toluenesulfonate), 1-naphthalenesulfonate, 2-naphthalenesulfonate, acetate, trifluoroacetate, malate, tartrate, citrate, lactate, oxalate, succinate, fumarate, maleate, benzoate, salicylate, phenylacetate, and mandelate.

3. The use of the compound according to claim 1 or 2, its stereoisomers, tautomers, or pharmaceutically acceptable salts thereof, characterized in that: Used for: (i) preparing indoleamine-2,3-dioxygenase inhibitors; (ii) preparing a medicament for preventing and / or treating an indoleamine-2,3-dioxygenase-mediated disease; or (iii) Preparation of anti-tumor drugs.

4. The use according to claim 3, characterized in that The indoleamine-2,3-dioxygenase-mediated disease is cancer, neurodegenerative disease, HIV infection, eye disease, psychological disorder, depression, anxiety, Alzheimer's disease and / or autoimmune disease.

5. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: the compound according to claim 1 or 2, its stereoisomers, tautomers, or pharmaceutically acceptable salts thereof; and a pharmaceutically acceptable carrier and other anti-tumor drugs, wherein the other anti-tumor drugs are selected from: chemotherapy drugs, targeted anti-tumor drugs and checkpoint protein antibodies.

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