HK1258589B - Spiro-condensed pyrrolidine derivatives as deubiquitylating enzymes (dub) inhibitors - Google Patents

Description

Spiro-condensed pyrrolidine derivatives as Deubiquitinase (DUB) inhibitors

The present invention relates to novel compounds and methods for the preparation of inhibitors of deubiquitinating enzymes (DUBs). In particular, the invention relates to the inhibition of Cezanne 1. The invention also relates to the use of DUB inhibitors in the treatment of cancer.

Background

The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.

Ubiquitin is a small protein consisting of 76 amino acids, which is important for the regulation of protein function in cells. Ubiquitination and deubiquitination are enzymatically mediated processes by which ubiquitination binds covalently or cleaves from a target protein. These processes are involved in the regulation of a number of cellular functions, including the regulation of cell cycle progression, apoptosis, cell surface receptor modification, DNA transcription and DNA repair. Thus, the ubiquitin system is involved in the pathogenesis of many disease states, including inflammation, viral infection, metabolic dysfunction, CNS disorders, and tumorigenesis.

Ubiquitin molecules are cleaved from proteins by Deubiquitinase (DUB), which is approximately 95 DUBs in human cells, and are classified into subfamilies based on sequence homology. The Ovarian Tumor (OTU) family consists of at least 14 active DUBs and is characterized by the presence of an OTU domain and a propensity to cleave ubiquitin chains in a ligation-specific manner. Cezanne1, also known as OTUD7B, is an 843 amino acid protein identified due to its similarity to OTU family member a20, which OTU family member a20 has shown a strong biochemical preference for K11 ubiquitin chain attachment.

Cezanne1 has been shown to act as a negative regulator of both the classical and non-classical NF-. kappa.B pathways. Cezanne1 has been shown to work on the canonical pathway by processing the RIP1 protein and the K63 chain on the non-canonical pathway via deubiquitination with the inhibitory component TRAF3(TNF receptor-related factor 3). It has also been shown to play a role in hypoxia by modulating HIF 1 α (hypoxia inducible factor 1 α) protein levels. The Cezanne1 siRNA reduced HIF 1 α protein levels under hypoxic conditions, and correspondingly reduced HIF 1 α target gene expression. Knockout of Cezanne1 results in higher levels of apoptosis following hypoxia. Because HIF 1 α has oncogenic properties and Cezanne1 has a pro-survival effect in hypoxia, Cezanne1 is considered a good target for pharmacological intervention.

Cezanne1 has been shown to promote T cell activation and inflammatory responses by modulating ZAP70 ubiquitination (Hu et al 2016 Journal of Experimental Medicine). This suggests that inhibition of Cezanne1 will result in a reduction of inflammatory responses. There is a continuing need for compounds that inhibit DUBs, such as Cezanne, for the treatment of inflammation.

Cezanne1 has been shown to play a role in cell proliferation, migration and invasion by antagonizing EGFR (epidermal growth factor receptor) uptake and degradation. Cezanne1 and Cezanne 2 were identified in gene screening to find DUB enzymes for EGFR. Cezanne1 overexpression results in higher levels of phosphorylated EGFR, lower levels of ubiquinated EGFR, and EGFR stabilization. In MDA-MB-231 breast cancer cells, knockdown of Cezanne1 resulted in reduced invasion and migration. Analysis of the cancer genomic map by Pareja et al (2012) showed that Cezanne1 is overexpressed in breast cancer, while amplification of the gene is seen in one third of breast tumors. The level of Cezanne1 expression correlates with poor prognosis.

Although some DUB inhibitors have been disclosed in the literature, there is a continuing need for compounds and pharmaceutical compositions that inhibit DUBs, such as Cezanne1 and USP30, for the treatment of cancer and other indications where DUB activity is observed.

Summary of The Invention

According to a first aspect of the present invention there is provided a compound of formula (I)

Or a pharmaceutically acceptable salt thereof, wherein:

R^1a，R^1b，R^1cand R^1dEach independently represents hydrogen or optionally substituted C₁-C₆Alkyl, or R^1aAnd R^1bTogether form an optionally substituted cycloalkyl ring, R^1cAnd R^1dTogether form an optionally substituted cycloalkyl ring, or R^1dAnd R^1eTogether form an optionally substituted cycloalkyl ring;

R^1eand R^1fEach independently represents hydrogen, fluorine, cyano, hydroxy, amino, optionally substituted C₁-C₆Alkyl, optionally substituted C₁-C₆Alkoxy or an optionally substituted 5-or 6-membered heteroaryl or aryl ring, or R^1eAnd R^1fOr R^1dTogether form an optionally substituted cycloalkyl ring;

ring a is a 5 to 11 membered monocyclic or bicyclic heterocycle which may optionally be further substituted.

In one aspect, the invention also relates to pharmaceutical compositions comprising a compound of the invention and one or more pharmaceutically acceptable excipients.

In another aspect, the compounds of the invention are useful for treating cancer.

Brief Description of Drawings

Figure 1 provides an image of Cezanne1 purified from mammalian cells. The FLAG-purified proteins or BSA at indicated concentrations were separated by SDS-PAGE and stained with Imperial protein stain (Pierce Biotechnology).

Figure 2 is a diagram showing the proteolytic activity of Cezanne1 as determined using fluorescence polarization assay. Different volumes of purified Cezanne1 as indicated were incubated with TAMRA-labeled peptides linked to ubiquitin via isopeptide bonds.

Detailed Description

The following definitions and explanations are for terms used throughout this document, including the specification and claims. Reference to a compound as described herein (e.g., a compound of formula I) includes reference to formula I and formula II, including embodiments of any sub-formulae thereof.

When any group of a compound of formula I is referred to as being optionally substituted, that group may be substituted or unsubstituted. The substitution may be substituted with one or more of the specified substituents, which may be the same or different. It will be understood that the number and nature of the substituents is selected to avoid any sterically undesirable combination.

In the context of this specification, unless otherwise indicated, an alkyl, alkylene, alkoxy, alkenyl, alkenylene or alkynyl substituent (or linker) group or alkyl, alkenyl moiety in a substituent may be straight or branched chain. The alkyl, alkylene and alkenyl chains may also include intervening heteroatoms, such as oxygen.

C_x-C_yAlkyl refers to a saturated aliphatic hydrocarbon group having x-y carbon atoms, which may be straight or branched. Example (b)Such as C₁-C₆The alkyl group contains 1 to 6 carbon atoms and includes C₁、C₂、C₃、 C₄、C₅And C₆. "branched" means that at least one carbon branch point is present on the group. For example, both tert-butyl and isopropyl are branched groups. C₁-C₆Examples of the alkyl group include methyl, ethyl, propyl, 2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-1-butyl, 2-methyl-3-butyl, 2-dimethyl-1-propyl, 2-methyl-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2-dimethyl-1-butyl, 3-dimethyl-1-butyl, 2-ethyl-1-butyl, 2-methyl-1-pentyl, 2-dimethyl-1-butyl, 2-methyl-1-pentyl, 2-dimethyl-1-pentyl, 2-methyl-pentyl, 2-pentyl, and, N-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl and n-hexyl. R^1a、R^1b、R^1c、R^1d、R^1e、 R^1f、R³、R⁴、R^4a、R⁶、R⁷、R^7a、Q¹Within the definition of (A) and R²C in the definition of the substituent(s)₁-C₆Alkyl radical, C₁-C₄Alkyl and C₁-C₃An alkyl group may be unsubstituted or substituted with one or more of the substituents defined herein. Thus, substituted C_1-C₆Examples of alkyl groups include CF₃、CH₂CF₃、CH₂CN、CH₂OH and CH₂CH₂OH。

C_x-C_yThe alkylene group or moiety may be straight or branched and refers to C as defined above_x-C_yA divalent hydrocarbon group having one less hydrogen atom in the alkyl group. C₁-C₆The alkylene group may include intervening heteroatoms, such as oxygen, and thus includes alkyleneoxy groups. The alkyleneoxy groups used herein also extend to embodiments in which a designated oxygen atom or a non-designated oxygen atom (e.g. a single oxygen atom) is located within the alkylene chain, e.g. CH₂CH₂OCH₂Or CH₂OCH₂。C₁-C₆Examples of alkylene groups include methylene, methyleneoxy, ethyleneEthylene oxide, n-propylene oxide, n-butylene oxide, methyl methylene and dimethyl methylene. Unless otherwise stated, R⁵、Q¹、Q^2a、Q^2bAnd Q^2cC within the definition of₁-C₆Alkylene radical, C₁-C₄Alkylene and C₁-C₃The alkylene group may be unsubstituted or substituted by one or more substituents as defined herein.

C₂-C₆Alkenyl means a straight or branched chain hydrocarbon group containing at least two carbon atoms and at least one double bond, and includes C₂-C₄An alkenyl group. Examples of alkenyl groups include ethenyl, propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1-hexenyl, 2-methyl-1-propenyl, 1, 2-butadienyl, 1, 3-pentadienyl, 1, 4-pentadienyl, and 1-hexadienyl. Unless otherwise stated, Q¹Within the definition of (A) and R²C in the definition of the substituent(s)₂-C₆An alkenyl group may be unsubstituted or substituted with one or more substituents as defined herein.

C₂-C₆Alkenylene means C as defined above₂-C₆A straight or branched hydrocarbon chain radical having one less hydrogen atom in the alkenyl radical. C₂-C₆Examples of alkenylene groups include vinylene, propenylene, and butenylene groups. Unless otherwise stated, for Q¹、Q^2a、Q^2bAnd Q^2cC in the definition of the substituent(s)₂-C₆Alkenylene groups may be unsubstituted or substituted with one or more substituents as defined herein.

C₂-C₆Alkynyl refers to a straight or branched hydrocarbon chain group containing at least two carbon atoms and at least one triple bond. Examples of alkenyl groups include ethynyl, propynyl, 2-propynyl, 1-butynyl, 2-butynyl and 1-hexynyl. Unless otherwise specified, Q¹Within the definition of (A) and R²C in the definition of the substituent(s)₂-C₆An alkynyl group may be unsubstituted or substituted by one or more of the substituents defined hereinAnd (4) substitution.

C₁-C₆Alkoxy means according to C above_x-C_yAlkyl is defined as having-O-C_x-C_yA group or moiety of an alkyl group. C₁-C₆The alkoxy group contains 1 to 6 carbon atoms and includes C₁、C₂、C₃、C₄、 C₅And C₆。C₁-C₆Examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, butoxy, pentyloxy, and hexyloxy. Alkoxy as used herein also extends to embodiments wherein a designated oxygen atom or a non-designated oxygen atom (e.g., a single oxygen atom) is located within the alkyl chain, e.g., CH₂CH₂OCH₃Or CH₂OCH₃. Thus, an alkoxy group may be attached to the rest of the molecule through a carbon, such as-CH₂CH₂OCH₃Alternatively, the alkoxy group is attached to the rest of the molecule via oxygen, e.g. -OC_1-6An alkyl group. In one instance, the alkoxy group is attached to the remainder of the molecule through an oxygen, but the alkoxy group contains another oxygen atom, e.g., -OCH₂CH₂OCH₃. Unless otherwise specifically stated, R^1e、R^1f、Q¹Within the definition of (A) and R²C in the definition of the substituent(s)₁-C₆Alkoxy and C₁-C₃An alkoxy group may be unsubstituted or substituted by one or more substituents as defined herein. Thus substituted C₁-C₆Examples of alkoxy groups include OCF₃、 OCHF₂、OCH₂CF₃、CH₂CH₂OCH₃And CH₂CH₂OCH₂CH₃。

The term "halogen" or "halo" refers to a chlorine, bromine, fluorine or iodine atom, in particular a chlorine or fluorine atom.

The term "oxo" refers to ═ O.

The term "amino" means-NR 'R', wherein R 'and R' each independently represent hydrogen or C₁-C₃An alkyl group.

The term "acylAmino "means-C (O) NR 'R', wherein R 'and R' each independently represent hydrogen or C₁-C₃An alkyl group.

For the avoidance of doubt, it will be understood that R and R are disclosed in the present application²、R⁸Cycloalkyl, heterocyclyl, aryl and heteroaryl rings within the definition of ring a do not include any labile ring structure or, in the case of heteroaryl and heterocyclic systems, any O-O, O-S or S-S bond. The ring system may be monocyclic or bicyclic. Bicyclic ring systems include bridged, fused and spiro ring systems. If present, the substituent may be attached to any suitable ring atom, which may be a carbon atom or, in the case of heteroaryl and heterocyclic systems, a heteroatom. Substitutions on the phenyl ring may also include changes in the ring atoms at the carbon substitution positions. For example, substitution on the phenyl ring may include a change in the ring atoms from the positions of carbon substitution to nitrogen, resulting in a pyridine ring.

"cycloalkyl" refers to a monocyclic saturated or partially unsaturated non-aromatic ring in which all of the ring atoms are carbon and have the indicated number of ring atoms. E.g. C₃-C₁₀Cycloalkyl refers to a monocyclic or bicyclic hydrocarbon ring containing 3 to 10 carbon atoms. C₃-C₁₀Examples of cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and decahydronaphthyl. Bicycloalkyl includes bridged ring systems such as bicycloheptane and bicyclooctane. Unless otherwise specifically stated, R^1a、 R^1b、R^1c、R^1d、R^1e、R^1f、R²、R⁸Cycloalkyl within the definition of (a) may be unsubstituted or substituted by one or more of the substituents defined herein.

An "aryl" group/moiety refers to any monocyclic or bicyclic hydrocarbon group containing at least one aromatic group and having 5 to 10 carbon atom ring members. Examples of aryl groups include phenyl and naphthyl. The bicyclic ring can be a fused aromatic ring wherein both rings are aromatic, such as naphthyl. Preferred aryl groups are phenyl and naphthyl, more preferably phenyl. Unless otherwise specifically stated, R^1e、R^1f、R²、 R⁸An aryl group within the definition of (a) may be unsubstituted or substituted with one or more substituents as defined herein.

As used herein, "heteroaryl" refers to a polyunsaturated monocyclic or bicyclic 5-10 membered aromatic moiety comprising at least one and up to 5 heteroatoms, particularly 1,2 or 3 heteroatoms selected from N, O and S, and the remaining ring atoms being carbon atoms, in a stable combination known to those skilled in the art. The heteroaryl nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atoms are optionally quaternized. The heteroaryl ring may be a single aromatic ring or a fused bicyclic ring, wherein the bicyclic ring system may be aromatic, or one of the fused rings may be aromatic while the other is at least partially saturated. In one example, a bicyclic heteroaryl is a group in which the entire fused ring system is aromatic. The bicyclic heteroaryl group may have at least one heteroatom in either ring of the fused ring. For example, a bicyclic ring having an aromatic ring fused to a partially saturated ring may include at least one heteroatom in the aromatic ring or the partially saturated ring. The bicyclic ring may be connected to the group (which is a substituent) via a ring containing a heteroatom or a ring containing only carbon. The point of attachment of the heteroaryl group to the group of which it is a substituent may be via a carbon atom or a heteroatom (e.g. nitrogen). In the case where ring a is heteroaryl, the ring is aromatic and may be fused with another aromatic ring or partially saturated. Examples include pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, furyl, pyrrolyl, and the like,Oxazolyl, thiazolyl, pyrazolyl, triazolyl, tetrazolyl, indolyl, indolizinyl, isoindolyl, indolinyl, purinyl, furazanyl, imidazolyl, indazolyl, isothiazolylAzolyl group,A diazolyl group,Oxazinalkyl, tetrazolyl, thiadiazolyl, benzofuranyl, isobenzofuranyl, tetrahydrofuryl, benzothienyl, isobenzothiophenyl, benzimidazolyl, benzothiazolyl, naphthyridinyl, pteridinyl, pyrazinyl, 4H-quinolizinyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, imidazopyridinyl, pyrazolopyridyl, thiazolopyridyl, indolinyl, isoindolinyl, triazinyl, dihydropyridinyl, quinoxalinyl and dihydrobenzofuranylAn oxazine group. Unless otherwise specifically stated, R^1e、R^1f、R²、R⁸The heteroaryl groups within the definition of (a) may be unsubstituted or substituted by one or more substituents as defined herein.

As used herein, unless otherwise specified, "heterocyclyl" or "heterocyclic" when describing rings refers to a monocyclic saturated or partially unsaturated non-aromatic ring or a bicyclic saturated or partially unsaturated ring, wherein the bicyclic ring system is non-aromatic, the monocyclic or bicyclic ring having, for example, 3-10 members, wherein at least one member and up to 5 members, particularly 1,2 or 3 members, of the ring are heteroatoms selected from, for example, N, O and S, and the remaining ring atoms are carbon atoms, in stable combinations known to those skilled in the art. The heterocyclic nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atoms are optionally quaternized. As used herein, a heterocyclic ring may be a ring fused to another ring system to form a bicyclic ring, i.e., one or both of the heterocyclic carbons are common to the other ring system. The heterocyclic group of ring a is a 5 to 11-membered monocyclic or bicyclic ring. When ring a is bicyclic, the second ring (i.e., the moiety not containing-NH-c (o)) -may be aromatic, such as fused phenyl or pyridyl. When ring a is bicyclic, generally any further substituents will be on the second ring. Examples of heterocyclyl groups include azetidinyl, pyrrolidinyl, piperidinyl, azepinyl, diazepinyl, dihydrofuranyl (e.g., 2, 3-dihydrofuranyl, 2, 5-dihydrofuranyl), dioxolanyl, morpholinyl, piperidinyl, and piperidinyl,Oxazolidinyl, piperazinyl, tetrahydrofuranyl, thiomorpholinyl, dihydropyranyl (e.g. 3, 4-dihydropyranyl, 3, 6-dihydropyranyl), homopiperazinyl, dipiperazinylAlkyl, hexahydropyrimidyl, pyrazolinyl, pyrazolidinyl, 4H-quinolizinyl, quinuclidinyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothienyl, thiazolidinyl, benzopyranyl, tetrahydroquinolinyl, dihydrobenzoOxazinyl and tetrahydroisoquinolinyl. Examples of heterocyclyl rings A include piperidin-2-one, indolin-2-one, piperazin-2-one, pyrrolidin-2-one, 3, 4-dihydroquinolin-2 (1H) -one, 1H-pyrido [2,3-b ]][1，4]Oxazin-2 (3H) -ones, 3, 4-dihydropyrido [2,3-b ]]Pyrazin-2 (1H) -ones, 1, 5-dihydrobenzo [ e][1，4]Oxazepin-2 (3H) -ones, 3, 4-dihydro-1, 5-naphthyridin-2 (1H) -ones, 3, 4-dihydro-1, 6-naphthyridin-2 (1H) -ones, 3, 4-dihydro-1, 7-naphthyridin-2 (1H) -ones, 3, 4-dihydro-1, 8-naphthyridin-2 (1H) -ones and 3, 4-dihydropyrazino [2,3-b ] -ones]Pyrazin-2 (1H) -ones and 1,2,3, 5-tetrahydro-4H-pyrido [2,3-b][1，4]Diazepan-4-ones. Unless otherwise specifically stated, R²And R⁸The heterocyclyl group within the definition of (a) may be unsubstituted or substituted by one or more of the substituents defined herein. Examples of substituted heterocyclyl rings include 4, 5-dihydro-1H-maleimido, tetramethylene sulfoxide and hydantoinyl. The monocyclic or bicyclic heterocyclic ring a may optionally be further substituted as described in the specification.

"optionally substituted" as applied to any group means that the group may be substituted, if desired, with one or more substituents (e.g., 1,2,3, or 4 substituents), which may be the same or different.

At R^1a、R^1b、R^1c、R^1d、R^1e、R^1f、R³、R⁴、R^4a、R⁶、R⁷、R^7a、Q¹Within the definition of (A) and R²Within the definition of the substituents of (1), "substituted" and "optionally substituted" C₁-C₆Alkyl (including C)₁-C₄Alkyl radical, C₁-C₃Alkyl and C₁-C₂Alkyl) and C₁-C₆Alkoxy (including C)₁-C₄Alkoxy radical, C₁-C₃Alkoxy and C₁-C₂Alkoxy) and C₂-C₆Alkenyl (including C)₂-C₄Alkenyl) and C₂-C₆Alkynyl (including C)₂-C₄Alkynyl) examples of suitable substituents, and in R⁵、 Q¹、Q^2a、Q^2bAnd Q^2cWithin the definition of (A), C₁-C₆Alkylene (including C)₁-C₃Alkylene) and C₂-C₆Examples of suitable substituents for alkenylene include halogen, hydroxy, sulfhydryl, cyano, amino, nitro and SF₅(known mimics of nitro groups), in particular halogen (preferably fluorine or chlorine), hydroxy and cyano. Other suitable substituents include amido, C_1-3Alkylamino radical, C_2-6Alkenylamino, di-C₁-C₃Alkylamino radical, C₁-C₃Acylamino, di-C₁-C₃Acylamino, carboxyl, C₁-C₃Alkoxycarbonyl, carboxamido, carbamoyl, mono-C_1-3Carbamoyl, di-C_1-3Carbamoyl, in which any hydrocarbyl moiety may itself be substituted by halogen, e.g. fluorine, hydroxy, cyano, amino, nitro or SF₅(known mimics of nitro groups).

At R^1a、R^1b、R^1c、R^1d、R^1e、R^1f、R²、R⁸Within the definition of ring A, "substituted" and "optionallyExamples of suitable substituents for substituted "rings (i.e., cycloalkyl, heterocyclyl, aryl, and heteroaryl rings) include halogen, cyano, oxo, nitro, amino, hydroxy, C₁-C₆Alkyl or C₁-C₃Alkyl radical, C₁-C₆Alkoxy or C₁-C₃Alkoxy, aryl, heteroaryl, heterocyclyl, C₃-C₆Cycloalkyl, amino, C_1-3Alkylamino radical, C_2-6Alkenylamino, di-C₁-C₃Alkylamino radical, C₁-C₃Amido, di-C₁-C₃Amido, carboxyl, C₁-C₃Alkoxycarbonyl, carboxamido, carbamoyl, mono-C_1-3Carbamoyl, di-C_1-3Carbamoyl or the hydrocarbyl part of the above groups being itself substituted by halogen, e.g. fluorine, hydroxy, cyano, amino, nitro or SF₅And (4) substitution.

Examples of suitable substituents for the "substituted" and "optionally substituted" rings include, inter alia, fluorine, chlorine, oxo, cyano, C₁-C₃Alkyl radical, C₁-C₃Alkoxy, amino, amido, heterocyclyl, cycloalkyl, heteroaryl or aryl, wherein alkyl or alkoxy is optionally substituted with one or more (e.g. 1,2 or 3) substituents selected from halogen, hydroxy, sulfhydryl, cyano, amino, nitro and SF₅。

Substituents thus include, for example, Br, Cl, F, CN, Me, Et, Pr, tBu, OMe, OEt, OPr, C (CH)₃)₃、CH(CH₃)₂、CF₃、OCF₃、C(O)NHCH₃Cyclopropyl, phenyl, and the like. In the case of aryl, the substitution may be in the form of a ring from adjacent carbon atoms on the aryl ring, e.g. a cyclic acetal, e.g. O-CH₂-O。

In oxygen atom-containing groups such as hydroxyl and alkoxy, the oxygen atom may be replaced by sulfur to produce groups such as thio (SH) and thio-alkyl (S-alkyl). Optional substituents therefore include groups such as S-methyl. In a thio-alkyl group, the sulfur atom may be further oxidized to produce a sulfoxide or sulfone, and thusSelected substituents include groups such as S (O) -alkyl and S (O)₂-an alkyl group.

The term "treating" includes preventing and means ameliorating, alleviating the symptoms, temporarily or permanently eliminating the cause of the symptoms, or preventing or slowing the appearance of the symptoms of the disorder or condition in question. The compounds of the invention are useful for treating human and non-human animals.

The dose of the compound is an amount effective to prevent the onset of symptoms of the disorder or to treat some symptoms of the disorder in the patient. An "effective amount" or "therapeutically effective amount" or "effective dose" refers to an amount sufficient to elicit the desired pharmacological or therapeutic effect, resulting in an effective prevention or treatment of the disorder. Prevention of a disorder is manifested by delaying the onset of symptoms of the disorder to a medically significant degree. Treatment of a disorder is manifested by a reduction in the symptoms associated with the disorder or an improvement in the recurrence of the symptoms of the disorder.

Pharmaceutically acceptable salts of the compounds of the present invention include, but are not limited to, addition salts (e.g., phosphates, nitrates, sulfates, borates, acetates, maleates, citrates, fumarates, succinates, methanesulfonates, benzoates, salicylates, and hydrohalites); salts derived from organic bases (e.g., lithium, potassium, and sodium), amino acids (e.g., glycine, alanine, valine, leucine, isoleucine, cysteine, methionine, and proline), and inorganic bases (e.g., triethylamine, hydroxide, choline, thiamine, and N-N' -diacetylethylenediamine). Other pharmaceutically acceptable salts include ammonium salts, substituted ammonium salts, and aluminum salts. Additional pharmaceutically acceptable salts include quaternary ammonium salts of the compounds of the present invention.

General methods for producing salts are well known to those skilled in the art. Such salts may be formed by conventional means, for example by reacting the free acid or free base form of the compound with one or more equivalents of the appropriate acid or base, optionally in a solvent or in a medium in which the salt is insoluble, and then removing the solvent or medium using standard techniques (for example in vacuo, by freeze-drying or by filtration). Salts may also be prepared by exchange of a counter ion with a compound in the form of a salt with another counter ion, for example using a suitable ion exchange resin.

When the compounds of the invention are present in different enantiomeric and/or diastereomeric forms, the invention relates to these compounds prepared as mixtures of isomers or racemates, whether in optically pure form or in mixtures with other isomers. Enantiomers differ only in their ability to rotate plane-polarized light in opposite directions by equal amounts, and are denoted as (+)/(S) or (-)/(R) forms, respectively. Individual enantiomers or isomers may be prepared by methods known in the art, such as optical resolution of the product or intermediate (e.g., chiral chromatographic separation, such as chiral HPLC, or asymmetric synthetic methods). Similarly, if the compounds of the invention exist as alternative tautomeric forms, such as keto/enol, amide/imide, the invention relates to isolated individual tautomers as well as mixtures of tautomers in all ratios.

Isotope of carbon monoxide

The compounds described herein may contain one or more isotopic substitutions, and reference to a particular element includes within its scope all isotopes of that element. For example, reference to hydrogen includes within its scope¹H、²H, (D) and³h (T). Similarly, references to carbon and oxygen, respectively, are included within their scope¹²C、¹³C and¹⁴c and¹⁶o and¹⁸and O. Examples of isotopes include²H、³H、¹¹C、¹³C、¹⁴C、³⁶Cl、¹⁸F、¹²³I、¹²⁵I、¹³N、¹⁵N、¹⁵O、¹⁷O、¹⁸O、³²P and³⁵S。

in a similar manner, reference to a particular functional group also includes isotopic variations within its scope unless the context indicates otherwise. For example, reference to an alkyl group such as ethyl also encompasses variations in which one or more of the hydrogen atoms on the group is in the form of a deuterium or tritium isotope, e.g., as an ethyl group (a per-deuterated ethyl group) in which all five of the hydrogen atoms are in the form of deuterium isotopes.

Isotopes may be radioactive or non-radioactive. In one embodiment, the compound does not contain a radioisotope. Such compounds are preferably used for therapeutic purposes. However, in another embodiment, the compound may comprise one or more radioisotopes. Compounds containing such radioisotopes may be useful in diagnostic settings.

Certain isotopically-labeled compounds of formula (I), for example those incorporating a radioactive isotope, are useful in drug and/or substrate tissue distribution studies. In view of their ease of incorporation and manner of detection, the radioisotopes are³H and¹⁴c is particularly useful for this purpose. With heavier isotopes (i.e.²H) Substitution may provide certain therapeutic advantages resulting from greater metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements, and thus may be preferred in certain circumstances. Using positron-emitting isotopes (e.g. of the type¹¹C、¹⁸F、¹⁵O and¹³n) can be used for Positron Emission Tomography (PET) studies to examine receptor occupancy. Isotopically-labelled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying examples and preparations, using a suitable isotopically-labelled reagent in place of the non-labelled reagent previously used.

Crystalline and amorphous forms

The compounds of formula (I) may exist in crystalline or amorphous form, and some crystalline forms may exist in polymorphic forms, which are included in the scope of the present invention. Polymorphic forms of the compound of formula (I) may be characterized and distinguished using a number of conventional analytical techniques including, but not limited to, infrared spectroscopy, raman spectroscopy, X-ray powder diffraction, differential scanning calorimetry, thermogravimetric analysis and solid-state nuclear magnetic resonance.

Thus, in a further embodiment, the present invention provides a compound according to any of the described embodiments in crystalline form. The compound may be 50% to 100% crystalline, more specifically at least 50% crystalline or at least 60% crystalline or at least 70% crystalline or at least 80% crystalline or at least 90% crystalline or at least 95% crystalline or at least 98% crystalline or at least 99% crystalline or at least 99.5% crystalline or at least 99.9% crystalline, for example 100% crystalline. The compound may also be in amorphous form.

The invention described herein relates to all crystalline forms, solvates and hydrates of any of the disclosed compounds, wherever prepared. Any compound disclosed in this application is within the scope of having an acid or basic center, such as a formate or an amino group, and then all salt forms of the compound are included in this application. In the case of pharmaceutical use, salts should be considered as pharmaceutically acceptable salts.

The present invention relates to any solvate of a compound and its salts. Preferred solvates are those formed by incorporating non-toxic pharmaceutically acceptable solvent (hereinafter referred to as solvating solvent) molecules into the solid state structure (e.g., crystal structure) of the compounds of the present invention. Examples of such solvents include water, alcohols (e.g., ethanol, isopropanol, and butanol), and dimethylsulfoxide. Solvates may be prepared by recrystallization of the compounds of the invention with a solvent or a mixture of solvents containing a solvating solvent. Whether a solvate is formed in any given instance can be determined by analysis of crystals of the compound using well-known and standard techniques such as thermogravimetric analysis (TGE), Differential Scanning Calorimetry (DSC) and X-ray crystallography.

The solvate may be a stoichiometric or non-stoichiometric solvate. Particular solvates may be hydrates, and examples of hydrates include hemihydrate, monohydrate, and dihydrate. For a more detailed discussion of solvates and methods for their preparation and characterization, see Bryn et al, Solid-State Chemistry of Drugs, 2 nd edition, published by SSCI, Inc of West Lafayette, IN, USA, 1999, ISBN 0-967-.

The present invention relates to pharmaceutically functional derivatives of compounds as defined herein, including ester derivatives and/or derivatives having or providing the same biological function and/or activity as any related compound of the invention. Thus, for the purposes of the present invention, the term also includes prodrugs of the compounds defined herein.

The term "prodrug" of a related compound includes any compound that, following oral or parenteral administration, is metabolized in vivo in an experimentally detectable amount and forms the compound within a predetermined time, e.g., 6-24 hours (i.e., 1-4 times per day) at dosing intervals.

Prodrugs of a compound may be prepared by modifying functional groups present on the compound in such a way that, when such a prodrug is administered to a mammalian subject, the modification is cleaved. Modifications are typically effected by synthesizing the parent compound with prodrug substituents. Prodrugs include compounds wherein a hydroxy, amino, mercapto, carboxyl or carbonyl group, respectively, in the compound is bonded to any group that can be cleaved in vivo to regenerate the free hydroxy, amino, mercapto, carboxyl or carbonyl group.

Examples of prodrugs include, but are not limited to, esters and carbamates of hydroxyl functional groups, ester groups of carboxyl functional groups, N-acyl derivatives, and N-Mannich bases. General information on Prodrugs can be found, for example, in Bundegaard, H. "Design of produgs" p.1-92, Elsevier, New York-Oxford (1985).

The compounds of the invention may be metabolized in vivo. Metabolites of the compounds of formula (I) are also within the scope of the invention. The term "metabolite" refers to all molecules derived from any compound of the invention in a cell or organism, preferably a mammal. Preferably, the term relates to a molecule that is different from any molecule present in any such cell or organism under physiological conditions.

The treatment defined in this application may be applied as the sole therapy or may involve conventional surgery or radiotherapy or chemotherapy in addition to the compounds of the invention. In addition, the compounds of formula (I) may also be used in combination with existing therapeutic agents for the treatment of cancer-related disorders, including small molecule therapeutics or antibody-based therapeutics.

According to a first aspect of the present invention there is provided a compound of formula (I)

Or a pharmaceutically acceptable salt thereof, wherein:

R^1a，R^1b，R^1cand R^1dEach independently represents hydrogen or optionally substituted C₁-C₆Alkyl, or R^1aAnd R^1bTogether form an optionally substituted cycloalkyl ring, R^1cAnd R^1dTogether form an optionally substituted cycloalkyl ring, or R^1dAnd R^1eTogether form an optionally substituted cycloalkyl ring;

R^1eand R^1fEach independently represents hydrogen, fluorine, cyano, hydroxy, amino, optionally substituted C₁-C₆Alkyl, optionally substituted C₁-C₆Alkoxy or an optionally substituted 5-or 6-membered heteroaryl or aryl ring, or R^1eAnd R^1fOr R^1dTogether form an optionally substituted cycloalkyl ring;

ring a is a 5 to 11 membered monocyclic or bicyclic heterocycle which may optionally be further substituted.

R^1aMay represent hydrogen. R^1aCan represent C₁-C₆An alkyl group. R^1aMay represent hydrogen or C₁-C₃Alkyl groups, for example, methyl or ethyl. When R is^1aIs represented by C₁-C₆When alkyl, R^1b，R^1c，R^1d， R^1eAnd R^1fMay each represent hydrogen. The alkyl group may be unsubstituted or substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅And especially fluorine.

R^1bMay represent hydrogen. R^1bCan represent C₁-C₆An alkyl group. R^1bMay represent hydrogen or C₁-C₃Alkyl groups, for example, methyl or ethyl. When R is^1bIs represented by C₁-C₆When alkyl, R^1a，R^1c，R^1d， R^1eAnd R^1fMay each represent hydrogen. The alkyl group may be unsubstituted or substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅And especially fluorine.

R^1cMay represent hydrogen. R^1cCan represent C₁-C₆An alkyl group. R^1cMay represent hydrogen or C₁-C₃Alkyl groups, for example, methyl or ethyl. When R is^1cIs represented by C₁-C₆When alkyl, R^1a，R^1b，R^1d，R^1eAnd R^1fMay each represent hydrogen. The alkyl group may be unsubstituted or substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅And especially fluorine.

R^1dMay represent hydrogen. R^1bCan represent C₁-C₆An alkyl group. R^1dMay represent hydrogen or C₁-C₃Alkyl groups, for example, methyl or ethyl. When R is^1dIs represented by C₁-C₆When alkyl, R^1a，R^1b，R^1c， R^1eAnd R^1fMay each represent hydrogen. The alkyl group may be unsubstituted or substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅And especially fluorine.

Alternatively, R^1aAnd R^1bMay together form a cycloalkyl ring. Additionally or alternatively, but preferably alternatively, R^1cAnd R^1dMay together form a cycloalkyl ring. The cycloalkyl ring may contain 3,4, 5 or 6 atoms, especially 3 or 4 atoms. When R is^1aAnd R^1bTogether form C₃-C₆Cycloalkyl ring, R^1c，R^1d，R^1eAnd R^1fMay be hydrogen. When R is^1cAnd R^1dWhen taken together to form a cycloalkyl ring, R^1a，R^1b，R^1eAnd R^1fMay each be hydrogen.

R^1eMay represent hydrogen, fluorine, cyano, hydroxy, amino, optionally substituted C₁-C₆Alkyl, optionally substituted C₁-C₆Alkoxy or an optionally substituted 5-or 6-membered heteroaryl or aryl ring. The alkyl and alkoxy groups may be substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅. The heteroaryl or aryl ring may be unsubstituted or substituted with: halogen, cyano, nitro, amino, hydroxy, C₁-C₆Alkyl or C₁-C₃Alkyl radical, C₁-C₆Alkoxy or C₁-C₃Alkoxy, aryl, heteroaryl, heterocyclyl, C₃-C₆Cycloalkyl radical, C_1-3Alkylamino radical, C_2-6Alkenylamino, di-C₁-C₃Alkylamino radical, C₁-C₃Acylamino, di-C₁-C₃Acylamino, carboxyl, C₁-C₃Alkoxycarbonyl, carboxamido, carbamoyl, mono-C_1-3Carbamoyl, di-C_1-3Carbamoyl or hydrocarbyl moieties per se substituted by halogen, e.g. fluorine, hydroxy, cyano, amino, nitro or SF₅Substituted any of the above substituents. In particular, said heteroaryl or aryl ring may be substituted by halogen, cyano, amino, C₁-C₃Alkoxy radical, C₁-C₆Alkyl substitution. R^1eMay represent hydrogen, fluorine, unsubstituted or substituted C₁-C₃Alkyl or unsubstituted or substituted C₁-C₃An alkoxy group. R^1eMay represent hydrogen or methyl. R^1eMay represent hydrogen. R^1eMay represent fluorine. R^1eMay represent a methyl group. R^1eMay represent a methoxy group. R^1eCan represent CF₃。R^1eCan represent an OCF₃. When R is^1eRepresents fluorine, cyano, hydroxy, amino, optionally substituted C₁-C₆Alkyl or optionally substituted C₁-C₆Alkoxy or optionally substituted 5-or 6-membered heteroaryl or aryl ring, R^1a，R^1b，R^1c，R^1dAnd R^1fMay each represent hydrogen.

R^1fMay represent hydrogen, fluorine, cyano, hydroxy, amino, optionally substituted C₁-C₆Alkyl, optionally substituted C₁-C₆Alkoxy, or an optionally substituted 5 or 6 membered heteroaryl or aryl ring. The alkyl and alkoxy groups may be substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅. The heteroaryl or aryl ring may be unsubstituted or substituted with: halogen, cyano, oxo, nitro, amino, hydroxy, C₁-C₆Alkyl or C₁-C₃Alkyl radical, C₁-C₆Alkoxy or C₁-C₃Alkoxy, aryl, heteroaryl, heterocyclyl, C₃-C₆Cycloalkyl radical, C_1-3Alkylamino radical, C_2-6Alkenylamino, di-C₁-C₃Alkylamino radical, C₁-C₃Acylamino, di-C₁-C₃Acylamino, carboxyl, C₁-C₃Alkoxycarbonyl, carboxamido, carbamoyl, mono-C_1-3Carbamoyl, di-C_1-3Halogen as any of the above substituents wherein the carbamoyl or hydrocarbyl moiety is itself substituted, e.g. fluorine, hydroxy, cyano, amino, nitro or SF₅. In particular, said heteroaryl or aryl ring may be substituted by halogen, cyano, amino, C₁-C₃Alkoxy radical, C₁-C₆Alkyl substitution. R^1fMay represent hydrogen, fluorine, unsubstituted or substituted C₁-C₃Alkyl or unsubstituted or substituted C₁-C₃An alkoxy group. R^1fMay represent hydrogen or methyl. R^1fMay represent hydrogen. R^1fMay represent fluorine. R^1fMay represent a methyl group. R^1fMay represent a methoxy group. R^1fCan represent CF₃。R^1fCan represent an OCF₃. When R is^1fRepresents fluorine, cyano, hydroxy, amino, optionally substituted C₁-C₆Alkyl or optionally substituted C₁-C₆Alkoxy or optionally substituted 5-or 6-membered heteroaryl or aryl ring, R^1a，R^1b， R^1c，R^1dAnd R^1eMay each represent hydrogen.

When R is^1eIs hydrogen, R^1fMay represent hydrogen, fluorine, cyano, hydroxy, amino, optionally substituted C₁-C₆Alkyl, optionally substituted C₁-C₆Alkoxy, or an optionally substituted 5 or 6 membered heteroaryl or aryl ring.

Alternatively, R^1eAnd R^1fMay together form a cycloalkyl ring. Alternatively, R^1eAnd R^1dMay together form a cycloalkyl ring. The cycloalkyl ring may contain 3,4, 5 or 6 atoms, especially 3 or 4 atoms. When R is^1eAnd R^1fTogether form C₃-C₆Cycloalkyl ring, R^1a，R^1b， R^1cAnd R^1dMay be hydrogen. When R is^1eAnd R^1dTogether form C₃-C₆Cycloalkyl ring, R^1a，R^1b， R^1cAnd R^1fMay each be hydrogen.

R^1a，R^1b，R^1c，R^1d，R^1eAnd R^1fThe cycloalkyl ring in the definition of (1) may be unsubstituted or substituted by one or more substituents selected from the group consisting of halogen, cyano, oxo, nitro, amino, hydroxy, C₁-C₃Alkyl radical, C₁-C₃Alkoxy radical, C_1-3Alkylamino radical, C_2-6Alkenylamino radical, C₁-C₃Acylamino, carboxyl, C₁-C₃Alkoxycarbonyl, carboxamido, carbamoyl, mono-C_1-3Carbamoyl and di-C_1-3Carbamoyl, wherein any hydrocarbyl moiety may itself be substituted by one or more halogen, hydroxy, cyano, amino, nitro or SF₅Especially fluorine substituted. In particular, said cycloalkyl ring may be unsubstituted or substituted by one or two substituents selected from halogen, cyano, oxo, nitro, amino, hydroxy, C₁-C₃Alkyl and C₁-C₃Alkoxy, wherein said alkyl and alkoxy groups may be substituted by one or more halogen, especially fluorine.

The compound may be wherein R^1a，R^1b，R^1c，R^1d，R^1eAnd R^1fEach represents the form of hydrogen. In this case, the compound may have the formula:

or a pharmaceutically acceptable salt thereof, wherein ring a is a 5 to 11 membered monocyclic or bicyclic heterocycle, which may be optionally further substituted.

Ring a may be monocyclic or bicyclic. If the ring is bicyclic, the second ring (i.e., the ring not directly attached to the pyrrolidine ring) may be aromatic, saturated or may be partially saturated. Preferably, the second ring is aromatic.

Ring A represents a 5-to 11-membered (e.g. 5, 6, 7, 8, 9, 10 or 11-membered) heterocyclic ring which may optionally be further substituted by one or more (e.g. mono-, di-, tri-or four) -Q¹-(R²)_nAnd (4) substitution.

Ring A may represent a 5-or 6-membered heterocyclic ring which may optionally be further substituted by one or more (e.g. one, two, three or four) -Q¹(R²)_nAnd (4) substitution.

Alternatively, ring a may represent a 9, 10 or 11 membered fused bicyclic heterocycle which may optionally be further substituted by one or more (e.g. mono, di, tri or four) -Q¹(R²)_nAnd (4) substitution.

Ring a may include one or more additional (e.g., 1,2, or 3) heteroatoms in addition to the amide nitrogen, wherein the additional heteroatoms are independently selected from nitrogen, oxygen, and sulfur. In particular, ring a may further comprise one or more additional heteroatoms selected from nitrogen and oxygen. When ring a is bicyclic, the additional heteroatom may be in the first ring (i.e. the ring containing-NH-c (o)) -and/or the second ring (i.e. the part of the fused ring not containing-NH-c (o)) -ring.

Ring A may be selected from piperidin-2-one, indolin-2-one, piperazin-2-one, pyrrolidin-2-one, 3, 4-dihydroquinolin-2 (1H) -one, 1H-pyrido [2,3-b ]][1，4]Oxazin-2 (3H) -ones, 3, 4-dihydropyrido [2,3-b ]]Pyrazin-2 (1H) -ones, 1, 5-dihydrobenzo [ e][1，4]Oxazepin-2 (3H) -ones, 3, 4-dihydro-1, 5-naphthyridin-2 (1H) -ones, 3, 4-dihydro-1, 6-naphthyridin-2 (1H) -ones, 3, 4-dihydro-1, 7-naphthyridin-2 (1H) -ones, 3, 4-dihydro-1, 8-naphthyridin-2 (1H) -ones and 3, 4-dihydropyrazino [2,3-b ] -ones]Pyrazin-2 (1H) -ones and 1,2,3, 5-tetrahydro-4H-pyrido [2,3-b ]][1，4]Diazepan-4-ones.

More particularly, ring A is selected from pyrrolidin-2-one, piperazine-2-one, 3, 4-dihydroquinolin-2 (1H) -one, 1H-pyrido [2,3-b ]][1，4]Oxazin-2 (3H) -ones, 3, 4-dihydropyrido [2,3-b ]]Pyrazin-2 (1H) -ones, 1, 5-dihydrobenzo [ e][1，4]Oxazacycloheptatrien-2 (3H) -ones and 1,2,3, 5-tetrahydro-4H-pyrido [2,3-b ]][1，4]Diazepan-4-ones.

In all of the schemes described in this application, ring A may be further substituted by one or more-Q¹-(R²)_nSubstituted, wherein each occurrence of-Q¹-(R²)_nThe same or different, and wherein:

n is 0 or 1;

Q¹represents halogen, cyano, oxo, nitro, hydroxy, -SR³，-NR³R⁴， -CONR³R⁴，-NR³COR⁴，-NR³CONR⁴R^4a，-COR³，-C(O)OR³，-SO₂R³， -SO₂NR³R⁴，-NR³SO₂R⁴，NR³SO₂NR⁴R^4a，-NR³C(O)OR⁴Optionally substituted-C₁-C₆Alkyl, optionally substituted-C₁-C₆Alkoxy, optionally substituted-C₂-C₆Alkenyl, optionally substituted-C₂-C₆Alkynyl, covalent bond, oxygen atom, sulfur atom, -OR⁵-， -SO-，-SO₂-，-CO-，-C(O)O-，-C₀-C₃alkylene-CONR³-C₀-C₃Alkylene group, -C₀-C₃alkylene-NR³-C₀-C₃Alkylene group, -C₀-C₃alkylene-NR³CO-C₀-C₃Alkylene group, -C₀-C₃alkylene-NR³CONR⁴-C₀-C₃Alkylene, -SO₂NR³-，-NR³SO₂-， -NR³SO₂N⁴-，-NR³C(O)O-，-NR³C(O)OR⁵-, optionally substituted C₁-C₆Alkylene or optionally substituted-C₂-C₆An alkenylene group;

R³，R⁴and R^4aEach independently represents hydrogen or optionally substituted C₁-C₆An alkyl group; and is

R⁵Represents optionally substituted C₁-C₆An alkylene group.

When n is 1, R²Represents an optionally substituted 3-to 10-membered heterocyclic, cycloalkyl, heteroaryl or aryl ring (Q when n is 0)¹Presence and R²Absent).

Ring A may be further substituted by one, two, three or four-Q¹-(R²)_nSubstitution is an additional substitution in addition to the oxo substitution that forms part of the amide.

In particular, ring A may be further unsubstituted or may be further substituted by one, two or three-Q¹-(R²)_nAnd (4) substitution. Ring A may be further substituted by one or two-Q¹-(R²)_nAnd (4) substitution. Each occurrence of-Q¹-(R²)_nMay be the same or different. Alternatively, ring A may be further substituted by one-Q¹-(R²)_nAnd (4) substitution. Q¹，R²And n is as defined herein. In some cases, ring a may not be further substituted.

In certain embodiments, ring a is substituted with a further optionally substituted ring, i.e., ring a is substituted with one or more-Q¹-(R²)_n(ii) partial substitution, wherein for at least one of said moieties n is 1. In general, ring A is substituted by only one-Q¹-(R²)_nPartially substituted, where n is 1, i.e. ring a is substituted with only one ring, which may additionally be other non-ring substituents.

In all schemes described in this application, Q¹May be selected from halogen (e.g. fluorine, chlorine or bromine), cyano, oxo, nitro, hydroxy, -SR³(e.g. mercapto), -NR³R⁴(e.g. amino or N, N-dimethylamino), -CONR³R⁴(e.g., amido), -NR³COR⁴(N-acetyl group), -NR³CONR⁴R^4a，-COR³(e.g., acetyl), -C (O) OR³(e.g., methoxycarbonyl or ethoxycarbonyl), -SO₂R³(e.g., methylsulfonyl), -SO₂NR³R⁴(e.g., dimethylaminosulfonyl), -NR3^SO₂R⁴，NR³SO₂NR⁴R^4a，-NR³C(O)OR⁴Optionally substituted-C₁-C₄Alkyl (e.g. propyl, isobutyl or tert-butyl), optionally substituted C₁-C₂Alkyl (e.g. methyl or ethyl), optionally substituted-C₁-C₆Alkoxy, optionally substituted-C₂-C₆Alkenyl, optionally substituted-C₂-C₆Alkynyl, covalent bond, oxygen atom, sulfur atom, -OR⁵-， -SO-，-SO₂-，-CO-，-C(O)O-，-C₀-C₃alkylene-CONR³-C₀-C₃Alkylene group, -C₀-C₃alkylene-NR³-C₀-C₃Alkylene (e.g. methylamino), -C₀-C₃alkylene-NR³CO-C₀-C₃Alkylene radical, -NR³CONR⁴-，-SO₂NR³-，-NR³SO₂-， -NR³SO₂NR⁴-，-NR³C(O)O-，-NR³C(O)OR⁵-, optionally substituted C₁-C₄Alkylene (e.g. methylene or ethylene) or optionally substituted-C₂-C₄Alkenylene (e.g. vinyl), wherein R³，R⁴，R^4aAnd R⁵As defined above.

In one embodiment, Q¹Selected from halogen, cyano, oxo, nitro, hydroxy, -SR³，-NR³R⁴，-CONR³R⁴，-NR³COR⁴，-NR³CONR⁴R^4a，-COR³， -C(O)OR³，-SO₂R³，-SO₂NR³R⁴，-NR³SO₂R⁴，NR³SO₂NR⁴R^4a， -NR³C(O)OR⁴Optionally substituted-C₁-C₄Alkyl, optionally substituted C₁-C₂Alkyl, optionally substituted-C₁-C₆Alkoxy, optionally substituted-C₂-C₆Alkenyl, optionally substituted-C₂-C₆Alkynyl, covalent bond, oxygen atom, sulfur atom, -OR⁵-，-SO-，-SO₂-， -CO-，-C(O)O-，-CONR³-，-NR³-，-NR³CO-，-NR³CONR⁴-， -SO₂NR³-，-NR³SO₂-，-NR³SO₂NR⁴-，-NR³C(O)O-，-NR³C(O)OR⁵-, optionally substituted C₁-C₄Alkylene or optionally substituted-C₂-C₄Alkenylene radical, in which R is³， R⁴，R^4aAnd R⁵As defined above.

When n is 0, ring A may be further substituted with one or more (e.g., one, two, three, or four) Q¹Substituted by substituentThe substituents being independently selected from halogen (e.g. fluorine, chlorine or bromine), cyano, oxo, nitro, hydroxy, -SR³，-NR³R⁴，-CONR³R⁴，-NR³C(O)R⁴， -NR³C(O)NR⁴R^4a，-C(O)R³，-C(O)OR³，-SO₂R³，-SO₂NR³R⁴， -NR³SO₂R⁴，NR³SO₂NR⁴R^4a，-NR³C(O)OR⁴，-C₁-C₆Alkyl radical, -C₁-C₆Alkoxy radical, -C₂-C₆Alkenyl, or-C₂-C₆Alkynyl, wherein alkyl, alkoxy, alkenyl or alkynyl may be unsubstituted or substituted with one or more substituents selected from halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅And wherein R is³，R⁴，R^4aAnd R⁵As defined above.

In particular, when n is 0, Q¹Can represent oxo, methyl, ethyl, CF₃Methoxy, halogen (e.g. fluorine or chlorine), -C (O) NR³R⁴Wherein R is³And R⁴Each independently represents hydrogen or methyl.

In particular examples, n is 0 and ring a represents a 5 or 6 membered heterocyclic ring, optionally substituted with one or more (e.g. one, two, three or four) Q¹Substituted with substituents independently selected from halogen (e.g. fluoro or chloro), oxo, C₁-C₆Alkyl or C optionally substituted by one or more fluoro₁-C₃Alkyl radicals, e.g. CF₃。

Alternatively, n is 0 and ring a represents a 9 or 10 membered heterocyclic ring, optionally substituted with one or more (e.g. one, two, three or four) Q¹Substituted with substituents independently selected from halogen (e.g. fluorine or chlorine), C₁-C₆Alkyl or C₁-C₃Alkyl or C₁-C₆Alkoxy or C₁-C₃Alkoxy, wherein said alkyl or alkoxy is optionally substituted by one or moreSubstituted by fluorine, e.g. CF₃Or C (O) NR³R⁴Wherein R is³And R⁴Each independently represents hydrogen and methyl.

When n is 1, Q¹Is a covalent bond OR a linking group selected from an oxygen atom, a sulfur atom, -OR⁵-，-SO-，-SO₂-，-CO-，-C(O)O-，-C₀-C₃alkylene-CONR³-C₀-C₃Alkylene group, -C₀-C₃alkylene-NR³-C₀-C₃Alkylene group, -C₀-C₃alkylene-NR³CO-C₀-C₃Alkylene radical, -NR³CONR⁴-，-SO₂NR³-，-NR³SO₂-，-NR³SO₂NR⁴-， -NR³C(O)O-，-NR³C(O)OR⁵-，C₁-C₆Alkylene or-C₂-C₆Alkenylene, wherein said alkylene or alkenylene is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅And wherein R is³，R⁴And R⁵As defined above.

In particular, when n is 1, Q¹May be selected from covalent bonds OR linking groups selected from covalent bonds, oxygen atoms, sulfur atoms, -OR⁵-，-SO-，-SO₂-，-CO-，-C(O)O-， -CONR³-，-NR³-，-NR³CO-，-NR³CONR⁴-，-SO₂NR³-，-NR³SO₂-， -NR³SO₂NR⁴-，-NR³C(O)O-，-NR³C(O)OR⁵-，C₁-C₆Alkylene or-C₂-C₆Alkenylene, wherein said alkylene or alkenylene is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅And wherein R is³， R⁴And R⁵As defined above.

In particular, when n is 1, Q¹Is a covalent bond or C₁-C₆Alkylene radicals, e.g. C₁-C₃Alkylene, wherein said alkylene is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅。

Preferably, ring A is substituted directly or via a linker by another ring, i.e. ring A is substituted by at least one-Q¹-(R²)_nWherein n is 1.

In all schemes described in this application, R²Represents a 3 to 10 membered heterocyclic, cycloalkyl, heteroaryl or aryl ring. R²Can be selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, decahydronaphthyl, phenyl, naphthyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, furyl, pyrrolyl,oxazolyl, thiazolyl, pyrazolyl, tetrazolyl, indolyl, indolizinyl, isoindolyl, indolinyl, purinyl, furazanyl, imidazolyl, indazolyl, isothiazolylThe group of azolyl groups,oxadiazolyl, tetrazolyl, thiadiazolyl, benzofuranyl, isobenzofuranyl, benzothiophenyl, isobenzothiophenyl, benzimidazolyl, benzothiazolyl, naphthyridinyl, pteridinyl, pyrazinyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, imidazopyridinyl, pyrazolopyridyl, thiazolopyridyl, isoindolinyl, triazinyl, dihydropyridinyl, quinoxalinyl, azetidinyl, pyrrolidinyl, piperidinyl, azepanyl, diazepanyl, dihydrofuranyl (e.g., 2, 3-dihydrofuranyl, 2, 5-dihydrofuranyl), dioxolanyl, morpholinyl,an oxazolidinyl group, which is a group of,oxazinyl, indolinyl, isoindolinyl, piperazinyl, tetrahydrofuranyl, thiomorpholinyl, dihydropyranyl (e.g., 3, 4-dihydropyranyl, 3, 6-dihydropyranyl), homopiperazinyl, dipiperazinylAlkyl, hexahydropyrimidyl, pyrazolinyl, pyrazolidinyl, 4H-quinolizinyl, quinuclidinyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothienyl, thiazolidinyl, benzopyranyl, tetrahydroquinolinyl, dihydrobenzoOxazinyl and tetrahydroisoquinolinyl.

R²May represent an optionally substituted 5-or 6-membered monocyclic heterocycle, cycloalkyl, heteroaryl or aryl ring.

Alternatively, R²May represent an optionally substituted 9-or 10-membered bicyclic heterocycle, cycloalkyl, heteroaryl or aryl ring.

In particular, R²Selected from phenyl, pyrazolyl, indazolyl, pyridyl, benzothiazolyl and pyrimidinyl. More particularly, R²Is phenyl.

In all schemes described in this application, R²May be optionally substituted with one or more substituents selected from halogen, cyano, oxo, nitro, hydroxy-SR⁶，-NR⁶R⁷， -CONR⁶R⁷，-NR⁶COR⁷，-NR⁶CONR⁷R^7a，-COR⁶，-C(O)OR⁶，-SO₂R⁶， -SO₂NR⁶R⁷，-NR⁶SO₂R⁷，NR⁶SO₂NR⁷R7^a，-NR⁶C(O)OR⁷，-C₁-C₆Alkyl radical, -C₁-C₆Alkoxy radical, -C₂-C₆Alkenyl, -C₂-C₆，-Q^2a-R⁸，-Q^2b-NR⁶CONR⁷R^7a， -Q^2b-NR⁶CONR⁷-Q^2c-R⁸，-Q^2b-NR⁶R⁷，-Q^2b-NR⁶-Q^2c-R⁸，-Q^2b-COR⁶， -Q^2b-CO-Q^2c-R⁸，-Q^2b-NR⁶COR⁷，-Q^2b-NR⁶CO-Q^2c-R⁸，-Q^2b-NR⁶C(O)OR⁷， -Q^2b-NR⁶C(O)O-Q^2c-R⁸，-Q^2b-SO₂R⁶，-Q^2b-SO₂-Q^2c-R⁸，-Q^2b-CONR⁶R⁷， -Q^2b-CONR⁶-Q^2c-R⁸，-Q^2b-CO₂R⁶，-Q^2b-CO₂-Q^2c-R⁸，-Q^2b-SO₂NR⁶R⁷， -Q^2b-SO₂NR⁶-Q^2c-R⁸，-Q²-NR⁶SO₂R⁷，-Q²-NR⁶SO₂-Q^2c-R⁸， -Q^2b-NR⁶SO₂NR⁷R^7aand-Q^2b-NR⁶SO₂NR⁷-Q^2c-R⁸Wherein said alkyl, alkoxy, alkenyl or alkynyl is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅Wherein

Q^2aRepresents a covalent bond, an oxygen atom, a sulfur atom, -SO-, -SO-₂-, -CO-, optionally substituted C₁-C₆Alkylene or optionally substituted C₂-C₆An alkenylene group;

Q^2band Q^2cEach independently represents a covalent bond, optionally substituted C₁-C₆Alkylene or optionally substituted C₂-C₆An alkenylene group;

R⁶，R⁷and R^7aEach independently represents hydrogen or optionally substituted C₁-C₆An alkyl group; and

R⁸represents an optionally substituted heterocyclic group, an optionally substituted heteroaryl group, an optionally substituted aryl group, or an optionally substituted cycloalkyl group.

R²May be substituted by one or more (e.g. one, two, three or four), especially one or two substituents independently selected from halogen, cyano, oxo, nitro, hydroxy, -SR⁶，-NR⁶R⁷，-CONR⁶R⁷，-NR⁶COR⁷，-NR⁶CONR⁷R^7a，-COR⁶， -C(O)OR⁶，-SO₂R⁷，-SO₂NR⁶R⁷，-NR⁶SO₂R⁷，NR⁶SO₂NR⁷R^7a， -NR⁶C(O)OR⁷，-C₁-C₆Alkyl radical, -C₁-C₆Alkoxy radical, -C₂-C₆Alkenyl, -C₂-C₆Alkynyl, -Q^2b-NR⁶CONR⁷R^7a，-Q^2b-NR⁶R⁷，-Q^2b-C(O)R⁶，-Q^2b-NR⁶C(O)R⁷， -Q^2b-NR⁶C(O)OR⁷，-Q^2b-SO₂R⁶，Q^2b-C(O)NR⁶R⁷，-Q^2b-CO₂R⁶， -Q^2b-SO₂NR⁶R⁷，-Q^2b-NR⁶SO₂R⁷and-Q^2b-NR⁶SO₂NR⁷R^7aWherein Q is^2bRepresents a covalent bond, optionally substituted C₁-C₆Alkylene or optionally substituted C₂-C₆Alkenylene, and wherein R⁶，R⁷And R^7aEach independently represents hydrogen or optionally substituted C₁-C₆An alkyl group, any of which is,alkoxy, alkenyl, alkynyl, alkylene or alkenylene is optionally substituted with one or more (e.g., mono-, di-, tri-or tetra-) substituents selected from halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅。

In particular, R²May be substituted by one or more substituents selected from halogen (e.g. fluorine), cyano, oxo, C₁-C₃Alkyl or C₁-C₃Alkoxy, wherein said alkyl or alkoxy is optionally fluoro, -CONR⁶R⁷，-NR⁶COR⁷，-Q^2a-R⁸，-Q^2b-NR⁶SO₂-Q^2c-R⁸Is substituted, wherein Q^2aIs a covalent bond, an oxygen atom, -CO-, -SO₂-or-C₁-C₃Alkylene radicals, Q^2bIs a covalent bond or C₁-C₃Alkylene and Q^2cIs a covalent bond, and wherein R⁶And R⁷Each independently selected from hydrogen or C₁-C₃Alkyl, and R⁸Is a 3-to 10-membered optionally substituted cycloalkyl, heterocyclyl, aryl or heteroaryl ring, especially a 3-to 6-membered monocyclic cycloalkyl, heterocyclyl, heteroaryl or aryl ring. More particularly, R⁸Selected from phenyl, piperazinyl, cyclopropyl, morpholinyl and piperidinyl.

More particularly, R²May be substituted by one or more substituents selected from halogen (e.g. chloro or fluoro), cyano, oxo, methyl, isopropyl, OMe, OCF₃O-isopropyl, -C (O) NHMe, -C (O) N (CH)₃)₂-NHC (O) Me, piperidinyl, -NHSO₂-cyclopropyl, Q^2a-phenyl, wherein Q^2aIs a covalent bond, an oxygen atom or a methyleneoxy group, Q^2a-piperazinyl, wherein Q^2aIs a covalent bond or-CO-and Q^2a-morpholinyl, wherein Q^2ais-CO-or-SO₂-。

In particular, R²Unsubstituted, mono-substituted or di-substituted.

In some cases, R²Optionally substituted by 3-to 10-membered heterocyclic, cycloalkyl, heteroaryl or aryl ringsEither directly or via a linker. The linking group may be an oxygen atom, a carbonyl group, -SO₂-，-NHSO₂-, or optionally substituted C₁-C₃An alkylene group. The linking group may be an oxygen, -CO-, or alkylene chain, for example, methylene or methyleneoxy. For example, R²May be substituted with a 5 or 6 membered ring selected from phenyl, piperidinyl, piperazinyl and morpholinyl. In addition to the ring substitution, R²May be further substituted with one or more non-cyclic substituents selected from the group consisting of halogen, cyano, oxo, C₁-C₃Alkyl radical, C₁-C₃Alkoxy, wherein said alkyl or alkoxy may optionally be fluoro, -C (O) NHMe, -C (O) N (CH)₃)₂and-NHC (O) Me substitution.

In some cases, R²Represents a 3-to 10-membered heterocyclic, cycloalkyl, heteroaryl or aryl ring selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, decahydronaphthyl, phenyl, naphthyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, furanyl, pyrrolyl,oxazolyl, thiazolyl, pyrazolyl, tetrazolyl, indolyl, indolizinyl, isoindolyl, indolinyl, purinyl, furazanyl, imidazolyl, indazolyl, isothiazolylThe group of azolyl groups,oxadiazolyl, tetrazolyl, thiadiazolyl, benzofuranyl, isobenzofuranyl, benzothiophenyl, isobenzothiophenyl, benzimidazolyl, benzothiazolyl, naphthyridinyl, pteridinyl, pyrazinyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, imidazopyridinyl, pyrazolopyridyl, thiazolopyridyl, isoindolinyl, triazinyl, dihydropyridyl, quinoxalyl, azetidinyl, pyrrolidinyl, piperidinyl,azepane, diazepanyl, dihydrofuranyl (e.g., 2, 3-dihydrofuranyl, 2, 5-dihydrofuranyl), dioxolanyl, morpholinyl,an oxazolidinyl group, which is a group of,oxazinyl, indolinyl, isoindolinyl, piperazinyl, tetrahydrofuranyl, thiomorpholinyl, dihydropyranyl (e.g., 3, 4-dihydropyranyl, 3, 6-dihydropyranyl), homopiperazinyl, dipiperazinylAlkyl, hexahydropyrimidyl, pyrazolinyl, pyrazolidinyl, 4H-quinolizinyl, quinuclidinyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothienyl, thiazolidinyl, benzopyranyl, tetrahydroquinolinyl, dihydrobenzoOxazinyl and tetrahydroisoquinolinyl unsubstituted or substituted with one or more (e.g., one, two or three) substituents selected from halo (e.g., fluoro or chloro), cyano, oxo, nitro, hydroxy, -SR⁶，-NR⁶R⁷，-CONR⁶R⁷， -NR⁶COR⁷，-NR⁶CONR⁷R^7a，-COR⁶，-C(O)OR⁶，-SO₂R⁶，-SO₂NR⁶R⁷， -NR⁶SO₂R⁷，NR⁶SO₂NR⁷R^7a，-NR⁶C(O)OR⁷，-C₁-C₆Alkyl radical, -C₁-C₆Alkoxy radical, -C₂-C₆Alkenyl, -C₂-C₆Alkynyl, -Q^2a-R⁸，-Q^2b-NR6CONR⁷R^7a， -Q^2b-NR⁶CONR⁷-Q^2c-R⁸，-Q^2b-NR⁶R⁷，-Q^2b-NR⁶-Q^2c-R⁸，-Q^2b-COR⁶， -Q^2b-CO-Q^2c-R⁸，-Q^2b-NR⁶COR⁷，-Q^2b-NR⁶CO-Q^2c-R⁸， -Q^2b-NR⁶C(O)OR⁷，-Q^2b-NR⁶C(O)O-Q^2c-R⁸，-Q^2b-SO₂R⁶， -Q^2b-SO₂-Q^2c-R⁸，Q^2b-CONR⁶R⁷，-Q^2b-CONR⁶-Q^2c-R⁸，-Q^2b-CO₂R⁶， -Q^2b-CO₂-Q^2c-R⁸，-Q^2b-SO₂NR⁶R⁷，-Q^2b-SO₂NR⁶-Q^2c-R⁸， -Q^2b-NR⁶SO₂R⁷，-Q^2b-NR⁶SO₂-Q^2c-R⁸，-Q²-NR⁶SO₂NR⁷R⁸and-Q^2b-NR⁶SO₂NR⁷-Q^2c-R⁸Wherein said alkyl, alkoxy, alkenyl or alkynyl is optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, mercapto, cyano, amino, nitro and SF₅Wherein Q is^2aRepresents a covalent bond, an oxygen atom, a sulfur atom, -SO-, -SO-₂-，-CO-，C₁-C₆Alkylene or optionally substituted C₂-C₆Alkenylene radical, Q^2bAnd Q^2cEach independently represents a covalent bond, optionally substituted C₁-C₆Alkylene or optionally substituted C₂-C₆Alkenylene radical, R⁶，R⁷And R^7aEach independently represents hydrogen or optionally substituted C₁-C₆Alkyl, and R⁸Represents an optionally substituted heterocyclic group, an optionally substituted heteroaryl group, an optionally substituted aryl group, or an optionally substituted cycloalkyl group.

In particular, R²Can be selected from phenyl, pyrazolyl, indazolyl, pyridyl, benzothiazolyl and pyrimidinyl, wherein the ring is unsubstituted or substituted with one or more (e.g., one, two or three) substituents selected from halogen, cyano, oxo, C₁-C₃Alkyl or C₁-C₃Alkoxy, wherein said alkyl or alkoxy is optionally fluoro, -CONR⁶R⁷，-NR⁶COR⁷，-Q^2a-R⁸， -Q^2b-NR⁶SO₂-Q^2c-R⁸Is substituted, wherein Q^2aIs a covalent bond, an oxygen atom, -CO-, -SO₂-or-C₁-C₃Alkylene radicals, Q^2bIs a covalent bond or C₁-C₃Alkylene radicals, Q^2cIs a covalent bond, wherein R⁶And R⁷Each independently selected from hydrogen or C₁-C₃Alkyl and R⁸Is a 3-to 10-membered optionally substituted cycloalkyl, heterocyclyl, aryl or heteroaryl ring.

The present invention also relates to compounds of formula (I), or a pharmaceutically acceptable salt thereof, wherein:

R^1a，R^1b，R^1cand R^1dEach independently selected from hydrogen and C which may be optionally substituted by fluorine₁-C₃An alkyl group;

R^1eand R^1fEach independently selected from hydrogen, fluorine, C₁-C₃Alkyl or C₁-C₃Alkoxy, wherein said alkyl or alkoxy is optionally substituted with fluoro;

ring A is a monocyclic or bicyclic 5-to 10-membered heterocycle, which is optionally further substituted by one, two or three-Q¹-(R²)_nIs substituted, wherein Q¹，R²And n is as defined herein.

The present invention also relates to compounds of formula (I), or a pharmaceutically acceptable salt thereof, wherein:

R^1a，R^1b，R^1c，R^1d，R^1eand R^1fEach represents hydrogen;

ring A is selected from pyrrolidin-2-one, piperazine 2-one,3, 4-dihydroquinolin-2 (1H) -one, 1H-pyrido [2,3-b][1，4]Oxazin-2 (3H) -ones, 3, 4-dihydropyrido [2,3-b ]]Pyrazin-2 (1H) -ones, 1, 5-dihydrobenzo [ e][1，4]Oxazacycloheptatrien-2 (3H) -ones and 1,2,3, 5-tetrahydro-4H-pyrido [2,3-b ]][1，4]Diazepan-4-ones in which the ring is optionally further substituted by one, two or three-Q¹-(R²)_nIs substituted, wherein Q¹，R²And n is as defined herein.

The present invention also relates to compounds of formula (I), or a pharmaceutically acceptable salt thereof, wherein:

R^1a，R^1b，R^1c，R^1d，R^1eand R^1fEach represents hydrogen;

ring A represents a monocyclic or bicyclic 5-to 10-membered heterocycle, which is optionally further substituted by one, two or three-Q¹-(R²)_nSubstitution; wherein

n is 0 or 1;

R²selected from piperidyl, pyrrolyl, phenyl, pyrazolyl, isoOxazolyl, indazolyl, pyridyl, dihydropyridinyl, benzothiazolyl, and pyrimidinyl;

Q¹as defined in the present application.

Examples of the monocyclic and bicyclic heterocycles represented by a include those shown below:

wherein denotes a point of attachment to the rest of the molecule, i.e. to pyrrolidine cyanamide to form a spiro ring, and wherein the ring is optionally substituted by one or more-Q¹-(R²)_nAnd (4) substitution.

Examples of novel compounds of formula I include:

2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7 '-chloro-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7 '-methoxy-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (5-isopropyl-2-methoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7' - ([1,1' -biphenyl ] -4-yl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (4- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (2-fluoro-5-methylphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (3-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1-methyl-1H-pyrazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-7' - (4-phenoxyphenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile 7'- (1-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (4-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (2-chloro-5- (trifluoromethoxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

5- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -yl) -N-methylpyridine carboxamide

7'- (2- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -yl) -N-methylbenzamide

7'- (3- ((2-chlorobenzyl) oxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (4- (4-methylpiperazin-1-yl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (6-methoxypyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (5-fluoro-2-isopropoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (3-methyl-1H-indazol-6-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (4- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1-methyl-1H-indazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (5-methyl-1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) cyclopropanesulfonamide

7'- (3-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-7' - (pyrimidin-5-yl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide

3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) -N, N-dimethylbenzamide

N- (4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide

7'- (4- (Morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (3, 5-dimethyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (2-methylpyridin-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-7' - (3- (piperidin-1-yl) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (2- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide

7'- (4- (morpholine-4-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (3- (morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1-methyl-6-oxo-1, 6-dihydropyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (2-methylbenzo [ d ] thiazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-6' -phenyl-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (3-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4-fluorophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (3-fluorophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

1-cyano-N, N-dimethyl-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -carboxamide

1-cyano-N-methyl-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -carboxamide

2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (4-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (3-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (3-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

2-oxo-6-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

2-oxo-6- (trifluoromethyl) -1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

2-oxo-7-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

7- (4-cyanophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

7- (4-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

7- (3-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

3-oxo-3, 4-dihydro-1H-spiro [ pyrido [2,3-b ] pyrazine-2, 3 '-pyrrolidine ] -1' -carbonitrile

6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitriles

(R) -6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

(S) -2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(R) -2 '-oxo-6' -phenyl-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

(S) -2-oxo-7-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(S) -7- (3-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (4-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (3-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (4-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(S) -7- (3-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(S) -7- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2, 3-b)][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (3-cyanophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(8R) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

(8S) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

7, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

8-Ethyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

8-benzyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

8-methyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

2-oxo-1, 5-dihydro-2H-spiro [ benzo [ e ] [1,4] oxazepine-3, 3 '-pyrrolidine ] -1' -carbonitrile

2-oxo-1, 2,4, 5-tetrahydrospiro [ pyrido [2,3-b ] [1,4] diazepane-3, 3 '-pyrrolidine ] -1' -carbonitrile

8-methyl-7, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7 '-chloro-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7 '-methoxy-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (5-isopropyl-2-methoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7' - ([1,1' -biphenyl ] -4-yl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (4- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (2-fluoro-5-methylphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (3-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1-methyl-1H-pyrazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-7' - (4-phenoxyphenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (4-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (2-chloro-5- (trifluoromethoxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

5- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -yl) -N-methylpyridine carboxamide

7'- (2- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -yl) -N-methylbenzamide

7'- (3- ((2-chlorobenzyl) oxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (4- (4-methylpiperazin-1-yl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (6-methoxypyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (5-fluoro-2-isopropoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (3-methyl-1H-indazol-6-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (4- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1-methyl-1H-indazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (5-methyl-1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) cyclopropanesulfonamide

7'- (3-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-7' - (pyrimidin-5-yl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide

3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) -N, N-dimethylbenzamide

N- (4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide

7'- (4- (Morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (3, 5-dimethyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (2-methylpyridin-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-7' - (3- (piperidin-1-yl) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (2- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide

7'- (4- (morpholine-4-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (3- (morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1-methyl-6-oxo-1, 6-dihydropyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (2-methylbenzo [ d ] thiazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-6' -phenyl-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (3-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4-fluorophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (3-fluorophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

1-cyano-N, N-dimethyl-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -carboxamide

1-cyano-N-methyl-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -carboxamide

2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (4-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (3-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (3-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

2-oxo-6-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

2-oxo-6- (trifluoromethyl) -1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

2-oxo-7-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

7- (4-cyanophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

7- (4-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

7- (3-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

3-oxo-3, 4-dihydro-1H-spiro [ pyrido [2,3-b ] pyrazine-2, 3 '-pyrrolidine ] -1' -carbonitrile

6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitriles

(R) -6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

(S) -2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(R) -2 '-oxo-6' -phenyl-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

(S) -2-oxo-7-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(S) -7- (3-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (4-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (3-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (4-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(S) -7- (3-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(S) -7- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2, 3-b)][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (3-cyanophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(8R) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

(8S) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

7, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

8-Ethyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

8-benzyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

8-methyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

2-oxo-1, 5-dihydro-2H-spiro [ benzo [ e ] [1,4] oxazepine-3, 3 '-pyrrolidine ] -1' -carbonitrile

2-oxo-1, 2,4, 5-tetrahydrospiro [ pyrido [2,3-b ] [1,4] diazepane-3, 3 '-pyrrolidine ] -1' -carbonitrile

8-methyl-7, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

2-oxo-6-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

7- (5-methyl-1H-indazol-4-yl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

7- (1, 4-dimethyl-1H-pyrazol-5-yl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

(R) -7'- (5-methyl-1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

(R) -7'- (4- (4-methylpiperazin-1-yl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile 6'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

(R) -7'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

(S) -7'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

(R) -6'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

1' -cyano-N- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-6-carboxamides

2-oxo-6- (piperidine-1-carbonyl) -1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

6- (1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -6- (1H-indazol-4-yl)-2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -1' -cyano-N- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2, 3-b)][1，4]Oxazine-3, 3' -pyrrolidines]-6-carboxamides

1' -cyano-2-oxo-N-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-6-carboxamides

1' -cyano-N- (2-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-6-carboxamides

7- (1-methyl-1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(R) -7- (1-methyl-1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4] Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (1-methyl-1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4] Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (1- (2-hydroxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(R) -7- (1- (2-hydroxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (1- (2-hydroxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (1- (2-methoxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(R) -7- (1- (2-methoxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (1- (2-methoxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

7- (6-methoxy-2-methylpyridin-3-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(R) -7- (6-methoxy-2-methylpyridin-3-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

(S) -7- (6-methoxy-2-methylpyridin-3-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

2 '-oxo-7' - (3- (trifluoromethoxy) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) -N, N-dimethylbenzamide

7'- (3- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

7'- (1-methyl-1H-pyrrol-2-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6' - ([1,1' -biphenyl ] -4-yl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (1-methyl-1H-pyrazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-6' - (3- (trifluoromethoxy) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-6' - (4-phenoxyphenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile 6'- (1-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

5- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -6' -yl) -N-methylpyridine carboxamide

6'- (2- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -6' -yl) -N-methylbenzamide

6'- (5-isopropyl-2-methoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (3- ((2-chlorobenzyl) oxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (6-methoxypyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (5-fluoro-2-isopropoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (3-methyl-1H-indazol-6-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (1-methyl-1H-indazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (5-methyl-1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) phenyl) cyclopropanesulfonamide

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) -N, N-dimethylbenzamide

2 '-oxo-6' - (pyrimidin-5-yl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) phenyl) acetamide

N- (4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) phenyl) acetamide

6'- (3- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (1-methyl-1H-pyrrol-2-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4- (Morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (3, 5-dimethyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

2 '-oxo-6' - (3- (piperidin-1-yl) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N- (2- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) phenyl) acetamide

6'- (4- (morpholine-4-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (3- (morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (1-methyl-6-oxo-1, 6-dihydropyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (2-methylbenzo [ d ] thiazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6' - (3, 5-dimethyliso)Azol-4-yl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-1-carbonitriles

6'- (2-chloro-5- (trifluoromethoxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4- (4-methylpiperazin-1-yl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

N-benzyl-4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) benzamide

6'- (3-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

6'- (4- (Morpholinylmethyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) -N, N-dimethylbenzamide, and

6'- (2-methylpyridin-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile.

It should be noted that each of the compounds listed above represent a particular and independent aspect of the present invention.

According to another aspect of the present invention, there is provided a process for preparing a compound of formula I, or a pharmaceutically acceptable salt thereof, comprising the step of reacting an amine of formula II with cyanogen bromide to form an N-CN compound:

wherein R is^1a-R^1fAnd a is as defined elsewhere in this application.

Another aspect of the invention provides pharmaceutical compositions comprising a compound of the invention.

The pharmaceutical compositions of the invention comprise any of the compounds of the invention in combination with any pharmaceutically acceptable carrier, adjuvant or vehicle.

Examples of pharmaceutically acceptable carriers are known to those skilled in the art and include, but are not limited to, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants and dispersants depending on the nature of the mode of administration and the dosage form. The compositions may be in the form of, for example, tablets, capsules, powders, granules, elixirs, lozenges, suppositories, syrups, and liquid preparations including suspensions and solutions. In the context of the present invention, the term "pharmaceutical composition" refers to a composition comprising an active agent and additionally comprising one or more pharmaceutically acceptable carriers. The compositions may also contain components selected from, for example, diluents, adjuvants, excipients, vehicles, preservatives, fillers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants and dispersants, depending on the nature of the mode of administration and the dosage form.

The compounds of the invention may be used for the treatment of disorders and diseases involving DUB inhibition, in particular inhibition of Cezanne1 and USP 30.

Another aspect of the invention provides a compound of formula (I) or a pharmaceutical composition thereof for use in therapy. In particular, the compounds of the invention have utility in the treatment of cancer, and more particularly in the treatment of cancer associated with DUB activity. The compounds of the invention may be useful for any DUB enzyme, including, but not limited to, Cezanne1 and USP 30.

The compounds described herein may be used in the preparation of medicaments for the treatment of cancers associated with DUB activity.

Another aspect of the invention provides a method of treating or preventing a cancer associated with the activity of Cezanne1 or USP30, the method comprising administering to an individual having a cancer associated with the activity of Cezanne1 or USP30 a pharmaceutically effective amount of a compound of the invention or a pharmaceutical composition thereof.

The compounds or compositions disclosed in the present application can be used to treat cancer. Reference to "cancer" or "tumor" includes, but is not limited to, breast cancer, ovarian cancer, prostate cancer, lung cancer, kidney cancer, stomach cancer, colon cancer, testicular cancer, head and neck cancer, pancreatic cancer, brain cancer, melanoma, bone cancer or other tissue organ cancers and blood cell cancers such as lymphoma and leukemia. Specific cancers include lymphoma, multiple myeloma, colorectal cancer and non-small cell lung cancer.

The compounds or compositions disclosed in the present application can be used to treat additional diseases associated with the activity of Cezanne 1.

A compound of the invention or a pharmaceutical composition thereof as described herein may be combined with one or more additional agents. The compounds may be combined with one or more additional anti-tumor therapeutic agents, such as chemotherapeutic drugs or other inhibitors of regulatory proteins. In one embodiment, the one or more anti-tumor therapeutic agents are chemotherapeutic agents. The chemotherapeutic agent may be selected from the group consisting of olaparide, mitomycin C, cisplatin, carboplatin, oxaliplatin, Ionizing Radiation (IR), camptothecin, irinotecan, topotecan, temozolomide, taxane, 5-fluoropyrimidine, gemcitabine, and doxorubicin. In another embodiment, the additional anti-tumor therapeutic is a BH-3 mimetic. In another embodiment, the BH-3 mimetic can be selected from one or more of, but is not limited to, ABT-737, ABT-199, ABT-263, and Obatocra (Obatocrax).

As mentioned above, inhibition of Cezanne1 will result in a reduction of the inflammatory response, and therefore the compounds of the invention (formula (I)) may be used in the treatment of inflammation.

As noted above, the compounds of the present invention may be used to treat disorders and diseases associated with inhibition of USP 30. The compounds of the invention may therefore be used for the treatment of disorders or diseases having elements involved in mitochondrial dysfunction.

Mitochondrial dysfunction is caused by defects in the mitochondria, a specialized compartment present in every cell of the body except red blood cells. When mitochondria fail, less and less energy is generated in cells and cell damage, and even cell death disappears. If this process is repeated throughout the body, the life of the subject undergoing this process can be severely impacted. Mitochondrial diseases occur most frequently in organs with high energy requirements, such as the brain, heart, liver, skeletal muscle, kidney and endocrine and respiratory systems.

Disorders involving mitochondrial dysfunction can be selected from disorders involving defects in mitochondrial autophagy, disorders involving mutations in mitochondrial DNA, disorders involving mitochondrial oxidative stress, disorders involving defects in mitochondrial membrane potential, mitochondrial biogenesis, disorders involving defects in mitochondrial shape or morphology, and disorders involving defects in lysosomal storage.

In particular, the condition involving mitochondrial dysfunction may be selected from neurodegenerative diseases; multiple Sclerosis (MS); mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like attack (MELAS) syndrome; leber's Hereditary Optic Neuropathy (LHON); cancer; neuropathy, ataxia, retinitis pigmentosa-maternal hereditary acute necrotizing encephalopathy (NARP-MILS); danon's disease; diabetes mellitus; diabetic nephropathy; metabolic disorders; heart failure; ischemic heart disease leading to myocardial infarction; psychosis, such as schizophrenia; multiple Sulfatase Deficiency (MSD); mucolipidosis ii (ml ii); mucolipidosis iii (ml iii); mucolipidosis iv (ml iv); GM 1-gangliosidosis (GM 1); neuronal ceroid lipid-lipofuscinosis (NCL 1); alpes disease; barth syndrome; beta-oxidation defects; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency syndrome; coenzyme Q10 deficiency; deficiency of Complex I; complex II deficiency; complex III deficiency; a deficiency of complex IV; complex V deficiency; COX deficiency; chronic progressive lateral ophthalmoplegic syndrome (CPEO); CPT I deficiency; CPT II deficiency; glutaric aciduria type II; Bass-Seldii syndrome; lactic acidosis; long-chain acyl-CoA dehydrogenase deficiency (LCHAD); a li's disease or syndrome; fatal infantile cardiomyopathy (LIC); left disease; glutaric aciduria type II; medium chain acyl-CoA dehydrogenase deficiency (MCAD); myoclonic epilepsy and fluffy-like red cellulose (MERRF) syndrome; mitochondrial cell disease; mitochondrial degenerative ataxia syndrome; mitochondrial DNA depletion syndrome; muscular neurogastrointestinal disorders and encephalopathy; pearson syndrome; pyruvate dehydrogenase deficiency; pyruvate carboxylase deficiency; POLG mutation; medium/short chain 3-hydroxyacyl-CoA dehydrogenase (M/SCHAD) deficiency; and very long chain acyl-CoA dehydrogenase (VLCAD) deficiency; and age-dependent decline in cognitive function and muscle strength.

The condition involving mitochondrial dysfunction may be a CNS disorder, such as a neurodegenerative disease. Neurodegenerative diseases include, but are not limited to, parkinson's disease, alzheimer's disease, Amyotrophic Lateral Sclerosis (ALS), huntington's disease, ischemia, stroke, Lewy body (Lewy) dementia, and frontotemporal dementia.

Dosage forms

For the treatment of mitochondrial dysfunction, the pharmaceutical composition of the present invention may be designed for administration by oral, parenteral or mucosal routes, and the choice or specific form of the composition depends on the route of administration. Thus, for oral administration, the compositions may be in the form of tablets, troches, lozenges, films, powders, elixirs, syrups, liquid preparations including dispersions, suspensions, emulsions, solutions or sprays, cachets, granules, capsules. For application to the mucosa, the composition may be in the form of a spray, inhalant, dispersion, suspension, emulsion, solution, gel, patch, film, ointment, cream, lotion, suppository, or the like. For parenteral administration, the compositions are in the form of liquid preparations such as solutions, dispersions, emulsions or suspensions, including liposomal compositions.

Formulations of the invention for parenteral administration include sterile water, aqueous-organic or organic solutions, suspensions and emulsions.

Such dosage forms may be prepared according to techniques well known in the art of pharmaceutical formulation. When a spray or an inhalant, the pharmaceutical composition may be administered nasally. Suitable formulations for this purpose are well known to those skilled in the art.

The pharmaceutical compositions of the present invention may also be in the form of suppositories for rectal administration. They are formulated such that the pharmaceutical composition is solid at room temperature and liquid at body temperature to allow release of the active compound.

The dosage may vary according to the requirements of the patient, the severity of the condition to be treated and the compound used. Determination of an appropriate dosage for a particular situation is within the purview of one skilled in the art. Typically, treatment is initiated at a smaller dose, which is less than the optimal dose of the compound. Thereafter, the dose is increased in small increments until the optimum effect in this case is achieved.

The magnitude of an effective dose of a compound will, of course, vary with the nature of the severity of the condition being treated and the particular compound and its route of administration. The selection of the appropriate dosage is within the ability of one of ordinary skill in the art without undue burden. Daily dosages range from about 10 μ g to about 100mg per kilogram body weight of the human and non-human animal, and typically from about 10 μ g to 30mg per kilogram body weight per dose. The above dose may be administered 1 to 3 times per day.

Synthesis method

The compounds of the present invention can be prepared by a variety of routes. Exemplary routes to certain compounds of the invention are shown below. Representative compounds of the invention can be synthesized according to the general synthetic methods described below, and are more particularly exemplified in the schemes below. As the scheme is an example, the invention should not be construed as being limited by the chemical reactions and conditions expressed. The preparation of the various starting materials used in the protocol is well within the skill of those in the art. Those skilled in the art will appreciate that the various transformations in the schemes may be accomplished in different orders, where appropriate. The following schemes describe general synthetic methods by which intermediates and target compounds of the present invention can be prepared. Intermediates prepared according to the general schemes and other materials, compounds and reagents known to those skilled in the art can be used to synthesize other representative compounds and stereoisomers, racemic mixtures, diastereomers and enantiomers thereof. All such compounds, stereoisomers, racemic mixtures, diastereomers and enantiomers thereof are intended to be included within the scope of the present invention.

All individual enantiomers listed were prepared from the corresponding racemic mixtures by chiral preparative HPLC or Supercritical Fluid Chromatography (SFC).

By liquid chromatography-mass spectrometry (LCMS) and/or¹H NMR characterized all compounds.

Abbreviations:

ABPR automatic back pressure regulating valve

AIBN azobisisobutyronitrile

Boc tert-butyloxycarbonyl group

br broad peak (NMR signal)

CAS chemical abstracts society

d double seam (NMR signal)

DCM dichloromethane

DIPEA diisopropylethylamine

DMF N, N-dimethylformamide

DMSO dimethyl sulfoxide

dppf 1,1' -bis (diphenylphosphino) ferrocene

ES electrospray ionization

EtOAc ethyl acetate

h hours

HATU 1- [ bis (dimethylamino) methylene]-1H-1, 2, 3-triazolo [4, 5-b]Pyridine compound3-oxidohexafluorophosphates

HPLC high performance liquid chromatography

IPA propan-2-ol

LCMS liquid chromatography-mass spectrometry

LiHMDS lithium bis (trimethylsilyl) amide

m multiplet (NMR signal)

MeCN acetonitrile

MeOH methanol

MTBE methyl tert-butyl ether

NBS N-bromosuccinimide

n-Bu n-butyl

NMR nuclear magnetic resonance

PE Petroleum Ether

Ph phenyl

Prep preparative

psi pounds per square inch

rt Room temperature

Retention time of RT

s singlet (NMR signal)

t triplet (NMR signal)

TBD 1,5, 7-triazabicyclo [4.4.0] dec 5-ene

TEA Triethylamine

TFA trifluoroacetic acid

THF tetrahydrofuran

TLC thin layer chromatography

LCMS method

Intermediate A3-amino-1-benzylpyrrolidine-3-carboxylic acid methyl ester

And a step a. To a stirred solution of methyl (tert-butoxycarbonyl) -L-serine ester (CAS number 2766-43-0; 30g, 136.9mmol) in DCM (600ml) was added pyridine (27.03g, 342.1mmol) at-50 ℃. Benzyl chloroformate (23.35g, 136.9mmol) was slowly added to the reaction mixture at-50 ℃ and the reaction mixture was stirred at rt for 16 hours. The resulting mixture was poured into 10% citric acid solution (1500ml), the organic phase was separated and the aqueous phase re-extracted with DCM (2 × 300 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (20% EtOAc in hexane) to give methyl O- ((benzyloxy) carbonyl) -N- (tert-butoxycarbonyl) -L-serine ester (22.0g, 62.323 mmol). LCMS: method 3, 4.98 min, MS: ES + 354.2;¹H NMR(400MHz， DMSO-d₆)δppm：7.49(d，J＝7.2Hz，1H)，7.36-7.39(m，5H)，5.16 (s，2H)，4.35-4.40(m，2H)，4.21-4.27(m，1H)，3.64(s，3H)， 1.38(s，9H)。

and b, step b. To a stirred solution of methyl O- ((benzyloxy) carbonyl) -N- (tert-butoxycarbonyl) -L-serine ester (22.0g, 62.3mmol) in DMF (150ml) at rt was added K₂CO₃(17.2g, 124.6mmol) and the reaction mixture was stirred at 65 ℃ for 1 hour. The mixture was cooled to rt, poured into water (2000ml) and extracted with EtOAc (2 × 500 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (5% EtOAc in hexane) to give methyl 2- ((tert-butoxycarbonyl) amino) acrylate (11.0g, 54.726 mmol).¹H NMR(400MHz，DMSO-d₆)δppm：8.39(s， 1H)，5.60(s，1H)，5.49(s，1H)，3.72(s，3H)，1.42(s，9H)。

And c, step (c). To a stirred solution of methyl 2- ((tert-butoxycarbonyl) amino) acrylate (11g, 54.726mmol) and N- (methoxymethyl) -N- (trimethylsilylmethyl) benzylamine (CAS number 93102-05-7; 12.97g, 54.7mmol) in DCM (250ml) was added TFA (0.3ml) at 0 ℃ under a nitrogen atmosphere. The reaction mixture was stirred at rt for 16 h. The resulting reaction mixture was diluted with DCM (200ml) and saturated NaHCO₃The solution (1500ml) was washed. The organic layer was separated and the aqueous layer re-extracted with DCM (2 × 200 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (5-20% EtOAc in hexane) to afford methyl 1-benzyl-3- ((tert-butoxycarbonyl) amino) pyrrolidine-3-carboxylate (9.0g, 26.9 mmol). LCMS: method 3, 4.69 min, MS: ES + 335.1;¹H NMR(400MHz，DMSO-d₆)δppm：7.59(s，1H)，7.28 (m，4H)，7.22-7.26(m，1H)，3.57-3.63(m，5H)，2.99(d，J＝10.0 Hz，1H)，2.68(d，J＝10.0Hz，1H)，2.58-2.61(m，1H)，2.47-2.49 (m，1H)，2.17-2.21(m，1H)，1.97-1.99(m，1H)，1.36(s，9H)。

and d, step d. To methyl 1-benzyl-3- ((tert-butoxycarbonyl) amino at 0 deg.C) To a stirred solution of pyrrolidine-3-carboxylic acid ester (0.8g, 2.39mmol) in DCM (15ml) was added TFA (4 ml). The reaction mixture was stirred at rt for 16 h. The reaction mixture was concentrated under reduced pressure and azeotropically distilled with DCM (2 × 20 ml). The resulting residue was dissolved in EtOAc (50mL) and washed with saturated NaHCO₃The solution (3 × 50ml) was washed. Separating the organic phase with Na₂SO₄Drying, filtration and concentration under reduced pressure gave methyl 3-amino-1-benzylpyrrolidine-3-carboxylate (0.57g, quantitative). This material was used directly in the next step without further purification. LCMS: method 1, 0.90 min, MS: ES + 235.3;¹H NMR(400MHz，DMSO-d₆)δppm：7.22-7.33 (m，5H)，3.63(s，3H)，3.58(s，2H)，2.88(d，J＝9.2Hz，1H)，2.68 (q，J＝7.2Hz，1H)，2.54-2.58(m，1H)，2.38(d，J＝9.6Hz，1H)， 2.21-2.25(m，1H)，1.61-1.68(m，1H)。

intermediate B tert-butyl 7 '-bromo-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carboxylate

Reagents and conditions: a) NBS, AIBN, CCl₄，76℃，16h b)LiHMDS， THF，-78℃，1h；rt，16h c)Fe，NH₄Cl, THF/water, 60 ℃, 16 h.

And a step a. At rt in N₂4-bromo-1-methyl-2-nitro-benzene (60g, 277mmol, 1.0eq) and NBS (59.3g, 333mmol, 1.2eq) were next charged to CCl₄To the mixture (600mL) was added AIBN (5.47g, 33.3mmol, 0.12 eq). The mixture was stirred at 76 ℃ for 16 hours. The reaction mixture was filtered and washed with 2M NaHCO₃The filtrate was washed (2X 250ml) and brine (400 ml) with Na₂SO₄Drying, filtration and concentration under reduced pressure gave a residue. The residue was purified by silica gel chromatography (0-5% EtOAc/PE) to give 4-bromo-1- (bromomethyl) -2-nitro-benzene (40g, 135mmol, 48.8% yield) as a yellow solid.¹H NMR(400MHz， CDCl₃)δppm：8.19(d，J＝1.6Hz，1H)，7.75(dd，J＝8.4，2.0Hz，1H)， 7.47(d，J＝8.4Hz，1H)，4.78(s，2H)。

And b, step b. At-78 ℃ in N₂LiHMDS (1M, 203ml, 1.5eq) was added dropwise to a mixture of pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (31.1g, 135mmol, 1.0eq) in THF (450 ml). The mixture was stirred at-78 ℃ for 30 minutes, then a solution of 4-bromo-1- (bromomethyl) -2-nitro-benzene (40g, 135mmol, 1.0eq) in THF (150ml) was added dropwise at-78 ℃. The mixture was stirred at-78 ℃ for 1h, then at rt for 16 h. LCMS showed the desired compound was detected. At rt by addition of saturated NH₄The reaction mixture was quenched with Cl solution (500ml) and then extracted with EtOAc (5 × 500 ml). The combined organic layers were washed with brine (2 × 1000ml) and Na₂SO₄Dried, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0-30% EtOAc/PE). To obtain 3- [ (4-bromo-2-nitro-phenyl) methyl]Pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (15g, 33.8mmol, 24.9% yield) as a yellow liquid.¹H NMR(400MHz，CDCl₃)δppm： 8.04(d，J＝10.0Hz，1H)，7.61-7.68(m，1H)，7.13(dd，J＝8.0，2.0Hz， 1H)，3.70-3.76(m，1H)，3.64(s，3H)，3.23-3.49(m，5H)，2.25-2.37(m， 1H)，1.78-1.92(m，1H)，1.46(s，9H)。

And c, step (c). To 3- [ (4-bromo-2-nitro-phenyl) methyl at 0 deg.C]1- (tert-butyl) 3-methyl pyrrolidine-1, 3-dicarboxylate (15g, 33.8mmol, 1.0eq) to a mixture of THF (250ml) and water (250ml) was added Fe (18.9g, 338mmol, 10.0eq) and NH₄Cl (18.1g, 338mmol, 11.8ml, 10.0 eq). The mixture was stirred at 60 ℃ for 16 hours. The reaction mixture was filtered, the filtrate diluted with water (50ml) and extracted with EtOAc (5 × 50 ml). The combined organic layers were washed with brine (2 × 60ml) and Na₂SO₄Drying, filtration and concentration under reduced pressure gave a residue. The residue was purified by silica gel chromatography (eluent 0-5% DCM/MeOH). To obtain 7 '-bromo-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline]Tert-butyl-1-carboxylate (12g, 31.4mmol, 93.0% yield) as a yellow solid.¹H NMR(400 MHz，DMSO-d6)δppm：10.34(s，1H)，7.07-7.16(m，2H)，7.03(s， 1H)，3.50-3.57(m，1H)，3.24-3.34(m，1H)，3.06(dd，J＝10.8，2.8Hz， 1H)，2.89(q，J＝11.6Hz，2H)，1.94-2.02(m，1H)，1.63-1.75(m，1H)， 1.37(s，9H)。

Intermediate C3-hydroxypyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester

Reagents and conditions: a) NaCN, NaHCO₃，Et₂O, water; b) HCl, 1, 4-bisAlkane, water; c) boc₂O，EtOAc，NaHCO₃(aq)；d)Cs₂CO₃，DMF；e)Fe， NH₄Cl, THF, water; f) TFA, DCM; g) CNBr, K₂CO₃，THF

And a step a. To a stirred solution of N-Boc-3-pyrrolidone (CAS No. 101385-93-7; 4.0g, 21.6mmol) in diethyl ether (50ml) and water (8ml) was added NaHCO in water (5ml) at 0 deg.C₃(3.6g, 43 mmol). NaCN (3.17g, 64.8mmol) was added to the reaction mixture at 0 ℃. The reaction mixture was stirred at rt for 24 hours. The resulting reaction mixture was poured into water (500ml) and extracted with ether (2 × 300 ml). The combined organic phases were washed with Na₂SO₄Drying, filtration and concentration under reduced pressure gave 3-cyano-3-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester (4.21g, 19.9 mmol). This material was used directly in the next step without further purification.¹H NMR(400MHz，DMSO-d6)δppm：6.90(s，1H)，3.48- 3.63(m，2H)，3.36-3.44(m，1H)，3.20-3.32(m，1H)，2.27- 2.33(m，1H)，2.13-2.20(m，1H)，1.42(s，9H)。

And b, step b. To a stirred solution of tert-butyl 3-cyano-3-hydroxypyrrolidine-1-carboxylate (4.2g, 19.8mmol) in MeOH (10.5ml) at 0 deg.C was added 1, 4-bisAlkane (42ml)) 4M HCl in (g). The reaction mixture was stirred at rt for 3 hours. The excess solvent was distilled under reduced pressure to give 3-hydroxypyrrolidine-3-carboxylic acid methyl ester HCl salt (4.2g, quantitative). This material was used directly in the next step without further purification. LCMS: method 3, 1.140 min, MS: ES + 146.07.

And c, step (c). To a stirred solution of 3-hydroxypyrrolidine-3-carboxylic acid methyl ester HCl salt (4.2g, 23.204 mmol) in EtOAc (42ml) at rt was added saturated NaHCO₃Solution (42 ml). Boc anhydride (10.12g, 46.4mmol) was added to the reaction mixture at rt. The reaction mixture was stirred at rt for 16 h. The resulting reaction mixture was poured into saturated NaHCO₃(200ml) and extracted with EtOAc (2X 100 rnl). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (30% EtOAc in hexane) to give 3-hydroxypyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (2.2g, 8.979 mmol). LCMS: method 1, 1.90 min, MS: ES + 246.2;¹H NMR(400 MHz，DMSO-d6)δppm：5.87(s，1H)，3.68(s，3H)，3.41-3.52(m， 2H)，3.28-3.32(M，2H)，2.09-2.18(m，1H)，1.91-1.94(m，1H)， 1.39(s，9H)。

intermediate D7-bromo-2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carboxylic acid tert-butyl ester

And a step a. To a stirred solution of 3-hydroxypyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (intermediate C; 0.5g, 2.04mmol) in THF (30ml) was added dropwise a solution of sodium bis (trimethylsilyl) amide (1M in THF; 2.04ml, 2.04mmol) at-78 ℃. The reaction mixture was stirred at-78 ℃ for 5 minutes. A solution of 2-fluoro-3-nitro-5-bromopyridine (CAS number 886372-98-1; 0.493g, 2.24mmol) in THF (1ml) was added to the reaction mixture at-78 deg.C. At-78 deg.C to-4 deg.CThe reaction mixture was stirred for 5 hours at 0 ℃. The resulting reaction mixture was quenched by slow addition of saturated ammonium chloride solution (20ml) at-40 ℃. The resulting reaction mixture was warmed to rt and combined with other three batches of the same scale prepared by the same method. The reaction mixture was diluted with water (50ml) and extracted with EtOAc (3 × 50 ml). The combined organic phases were separated and washed with brine (30 ml). Separating the organic phase with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (12% EtOAc in hexanes) to give 1- (tert-butyl) 3-methyl 3- ((5-bromo-3-nitropyridin-2-yl) oxy) pyrrolidine-1, 3-dicarboxylate (1.65g, 3.697 mmol). LCMS: method 1, 2.539 min, MS: ES +390.2, 392.2(M-2) (M-56);¹H NMR(400MHz，DMSO-d₆)δ ppm：8.78(s，1H)，8.63(s，1H)，4.00(d，J＝12.0Hz，1H)，3.70(d， J＝12.8Hz，1H)，3.65(s，3H)，3.48-3.54(m，1H)，3.37-3.46(m， 1H)，2.39-2.45(m，2H)，1.39(d，J＝6.4Hz，9H)。

and b, step b. To a stirred solution of 1- (tert-butyl) 3-methyl 3- ((5-bromo-3-nitropyridin-2-yl) oxy) pyrrolidine-1, 3-dicarboxylate (0.8g, 1.793mmol) in THF: water (1: 1; 8ml) at rt was added iron powder (1.0g, 17.927mmol) and ammonium chloride (0.957g, 17.93 mmol). The reaction mixture was heated at 70 ℃ for 18 hours. The resulting reaction mixture was cooled to rt and combined with another batch of the same scale prepared by the same method. The reaction mixture was filtered through celite hyflow. The celite bed was washed with EtOAc (100 ml). The combined filtrates were poured into water (50 ml). The organic phase was separated and the aqueous phase was re-extracted with EtOAc (3 × 50 ml). The combined organic phases were washed with brine (50 ml). The combined organic phases were separated over Na₂SO₄Drying, filtering and concentrating under reduced pressure to obtain 7-bromo-2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]An approximately 2: 3 mixture of tert-butyl (E) -1' -carboxylate and 1- (tert-butyl) 3-methyl 3- ((3-amino-5-bromopyridin-2-yl) oxy) pyrrolidine-1, 3-dicarboxylate(1.45g, quantitative). The resulting mixture was used in the next step without further purification. LCMS: method 1, 2.218 min, 2.429 min, MS: ES +328.0, 329.0(M +2) (M-56), 416.1, 418.1

And c, step (c). To 7-bromo-2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] at rt][1，4]Oxazine-3, 3' -pyrrolidines]To a stirred solution of an approximately 2: 3 mixture of tert-butyl (1' -carboxylate) and 3- ((3-amino-5-bromopyridin-2-yl) oxy) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (0.7g, 1.689mmol) in THF (20ml) was added 1,5, 7-triazabicyclo [4.4.0]Dec 5-ene (0.235g, 1.689 mmol). The reaction mixture was stirred at rt for 3 hours. The resulting reaction mixture was combined with another batch prepared by the same method on the same scale. The resulting reaction mixture was concentrated in vacuo and the residue was purified by flash chromatography (25% EtOAc in hexanes) to give 7-bromo-2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -carboxylic acid tert-butyl ester (1.3g, 3.618 mmol). LCMS: method 1, 2.125 min, MS: ES +328.0, 330.0(M +2) (M-56);¹H NMR(400MHz，DMSO-d₆) δppm：11.21(s，1H)，7.96(d，J＝2.0Hz，1H)，7.40(d，J＝2.4Hz，1 H)，3.68-3.75(m，1H)，3.52-3.60(m，2H)，3.64-3.44(m，1H)， 2.32-2.36(m，1H)，2.20-2.22(m，1H)，1.40(d，J＝10.0Hz，9H)。

intermediate E3-amino-1-benzylpyrrolidine-3-carboxylic acid methyl ester TFA salt

And a step a. To a solution of Boc-L-serine methyl ester (CAS number 2766-43-0; 30.0g, 136.9mmol) in DCM (300ml) was added pyridine (28.8ml, 342mmol) at-50 ℃ and stirred for 15 min. Benzyl chloroformate (25.67) at-50 deg.Cg, 150.5mmol) was added dropwise to the reaction mixture. The temperature of the reaction mixture was gradually increased to rt. The resulting reaction mixture was stirred at rt for 15 hours. After 15 h pyridine (22.0ml, 274mmol) and benzyl chloroformate (23.3g, 136.9mmol) were added at-50 ℃ and the reaction mixture was stirred at rt for 5 h. The resulting reaction mixture was quenched with 50% citric acid solution (500ml) and extracted with EtOAc (3 × 100 ml). The combined organic phases were collected and washed with saturated NaHCO₃The solution (50ml) was washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was triturated with N-hexane (200ml) to give O- ((benzyloxy) carbonyl) -N- (tert-butoxycarbonyl) -L-serine methyl ester (33.25g, 94.155 mmol). LCMS: method 3, 4.94 min, MS: ES + 354.1;¹H NMR(400 MHz，DMSO-d6)δppm：7.48(d，J＝7.2Hz，1H)，7.32-7.41(m，5H)， 5.14(s，2H)，4.36-4.40(m，2H)，4.23-4.26(m，1H)，3.63(s，3H)， 1.37(S，9H)。

and b, step b. To a solution of O- ((benzyloxy) carbonyl) -N- (tert-butoxycarbonyl) -L-serine methyl ester (33.0g, 93.48mmol) in DMF (330ml) at rt was added K₂CO₃(25.88g, 186.97 mmol). The reaction mixture was heated at 65 ℃ for 1 hour. The resulting reaction mixture was poured into water (1000ml) and extracted with EtOAc (3 × 150 ml). The combined organic phases were washed with brine solution (3 × 50ml) and Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (3% EtOAc in hexanes) to give methyl 2- ((tert-butoxycarbonyl) amino) acrylate (10.39g, 51.66 mmol). LCMS: method 1, 2.25 minutes.¹H NMR(400MHz，DMSO-d6)δppm：8.37(s，1H)，5.64(s，1H)，5.49 (s，1)，3.72(s，3H)，1.41(s，9H)。

And c, step (c). To a solution of 2- ((tert-butoxycarbonyl) amino) acrylate (10.3g, 51.24 mmol) in DCM (103ml) was added TFA (0.26ml) at 0 ℃. N- (methoxymethyl) -N- (trimethylsilylmethyl) -benzylamine (13.35g, 56.37mmol) was slowly added to the reaction mixture at 0 ℃. The reaction mixture was stirred at 0 ℃ for 15 minutes and then at rt for 15 hours. Unreacted starting material was observed after 15 hours, so N- (methyloxy) was again reacted at 0 deg.CMethyl) -N- (trimethylsilylmethyl) -benzylamine (3.64g, 15.373mmol) was added slowly to the reaction mixture. The reaction mixture was stirred at rt for an additional 15 hours. The resulting reaction mixture was poured into water (250ml) and Na was added₂CO₃Alkalizing. The resulting mixture was extracted with DCM (2 × 150ml) and the combined organic phases were washed with brine solution (50ml), Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (20% EtOAc in hexanes) to give methyl 1-benzyl-3- ((tert-butoxycarbonyl) amino) pyrrolidine-3-carboxylate (13.80g, 41.294 mmol). LCMS: method 3, 4.68 min, MS: ES + 335.3;¹H NMR(400MHz， DMSO-d6)δppm：7.59(s，1H)，7.21-7.33(m，5H)，3.57-3.62(m， 5H)，2.99(d，J＝10Hz，1H)，2.67(d，J＝10Hz，1H)，2.50-2.61(m， 2H)，2.14-2.18(m，1H)，1.98-1.99(m，1H)，1.35(s，9H)。

and d, step d. To a solution of methyl 1-benzyl-3- ((tert-butoxycarbonyl) amino) pyrrolidine-3-carboxylate (2.00g, 5.99mmol) in DCM (20ml) at rt was added TFA (2 ml). The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under reduced pressure and the resulting residue was triturated with ether (2 × 10ml) to give 3-amino-1-benzylpyrrolidine-3-carboxylic acid methyl ester TFA salt (2.50 g. This material was used directly in the next step without further purification. LCMS: method 1, 0.70 min, MS: ES + 235.4.

Intermediate F2- (4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazol-1-yl) ethan-1-ol

And a step a. To a solution of 4-bromo-1H-indazole (CAS number 186407-74-9; 1.000g, 5.076 mmol) in DMF (20ml) at rt was added 2-bromoethanol (0.43ml, 6.091mmol) and K₂CO₃(1.400g, 10.145 mmol). The reaction mixture was heated to 100 ℃ for 3 hours. The resulting mixture was poured into ice-cold water (50 ml). The resulting precipitate was collected by filtration, washed with hexane (50ml) and dried under high vacuumAnd (5) drying. By LCMS, the solid obtained contained a 2: 1 ratio of regioisomers. The desired product was isolated by column chromatography (23% EtOAc in hexanes) to give 2- (4-bromo-1H-indazol-1-yl) ethan-1-ol (0.700g, 2.904 mmol). LCMS: method 1, 1.629 min, MS: ES +241.20, 243.20;¹H NMR(400 MHz，DMSO-d6)δppm：8.03(s，1H)，7.71(d，J＝8.0Hz，1H)，7.28 -7.36(m，2H)，4.87(t，J＝5.6Hz，1H)，4.62(t，J＝5.2Hz，2H)， 3.78-3.82(m，2H)。

and b, step b. 2- (4-bromo-1H-indazol-1-yl) ethane-1-o 1(0.700g, 2.904mmol) at rt in 1, 4-bisTo a solution in an alkane (10ml) were added bis (pinacolato) diboron (1.102g, 4.356mmol) and KOAc (0.569g, 5.808 mmol). The reaction mixture was degassed at rt for 10 min, then PdCl was added₂(dppf) (0.212g, 0.290 mmol). The reaction mixture was heated at 100 ℃ for 1 hour. The resulting mixture was cooled to rt and concentrated under reduced pressure. The resulting residue was washed with n-hexane (10ml) to give 2- (4- (4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-indazol-1-yl) ethan-1-ol (1.0 g). LCMS: method 1, 1.880 min, MS: ES +289.50[ M +1 ]]. This material was used directly in the next step without further purification.

Intermediate G6-bromo-2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carboxylic acid tert-butyl ester

This intermediate was prepared in a manner analogous to that for intermediate D using 2, 6-dibromo-3-nitropyridine (CAS number 55304-80-8). LCMS: method 1, 1.892 min, MS: the number of bits in the ES + 384.40, 386.40,¹H NMR(400MHz，DMSO-d6)δppm：11.23(br s，1 H)，7.31(d，J＝8.0Hz，1H)，7.23(d，J＝8.0Hz，1H)，3.69-3.74(m， 1H)，3.52-3.62(m，3H)，2.33-2.39(m，1H)，2.21-2.24(m，1H)， 1.42(d，J＝10.4Hz，9H)。

scheme 1

Reagents and conditions: a) i) LiHMDS, hexane, THF; b) 10% Pd/C, H₂MeOH or Fe, NH₄Cl, THF, water; c) TFA, DCM; d) CNBr, K₂CO₃， THF

Example 12 ' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

Synthesis according to scheme 1

And a step a. To a stirred solution of pyrrolidine-1, 3-dicarboxylic acid tert-butyl methyl ester (CAS number 122684-33-7; 1.0g, 4.367mmol) in dry THF (15ml) at-78 deg.C was added a 1M solution of LiHMDS in hexane (1.08g, 6.55 mmol). The reaction mixture was stirred at-78 ℃ for 0.5 h, then 2-nitrobenzyl bromide (1.03g, 4.803mmol) was added at-78 ℃. The resulting reaction mixture was warmed to rt and stirred for 16 hours. The mixture was then poured into saturated ammonium chloride solution (20ml), diluted with water (100ml) and extracted with EtOAc (5 × 50 ml). The combined organic phases were dried over Na2SO4, filtered and concentrated under reduced pressure. The crude material obtained was purified by flash chromatography (6% EtOAc in hexane) to give 1- (tert-butyl) 3-methyl 3- (2-nitrobenzyl) pyrrolidine-1, 3-dicarboxylate (0.48g, 1.319 mmol). LCMS: method 1, 2.41 min, MS: ES + 365.4;¹H NMR(400MHz，DMSO-d6)δppm： 7.93(d，J＝8.0Hz，1H)，7.68(t，J＝7.2Hz，1H)，7.53(t，J＝8.0Hz， 1H)，7.36-7.41(m，1H)，3.57-3.64(m，1H)，3.34-3.42(m，6H)， 3.11-3.20(m，2H)，2.15-2.19(m，1H)，1.88-1.93(m，1H)，1.39 (s，9H)。

and b, step b. At rt to 3- (2-Nitro)To a stirred solution of 1- (tert-butyl) 3-methyl (0.2g, 0.549mmol) of phenylbenzyl) pyrrolidine-1, 3-dicarboxylic acid in MeOH (10ml) was added 10% dry Pd/C (0.2 g). H at rt₂The reaction mixture was purged with gas for 0.5 hours. The resulting reaction mixture was carefully filtered through celite hyflow and concentrated under reduced pressure to give 2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-1-carboxylic acid tert-butyl ester (0.14g, 0.463 mmol). LCMS: method 1, 2.20 min, MS: ES +247.4 (M-56);¹H NMR(400MHz， MeOD)δppm：7.19-7.24(m，2H)，7.02(t，J＝7.2Hz，1H)，6.90(d， J＝8.0Hz，1H)，3.71-3.76(m，2H)，3.49-3.69(m，2H)，2.93- 3.06(m，2H)，2.14-2.24(m，1H)，1.77-1.89(m，1H)1.47(s，9 H)。

and c, step (c). To a stirred solution of 2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carboxylic acid tert-butyl ester (0.13g, 0.43mmol) in DCM (1ml) was added TFA (1ml) at rt. The reaction mixture was stirred at rt for 2 hours. The resulting reaction mixture was concentrated under reduced pressure to give 1',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinolin ] -2' -one TFA salt (0.09g, 0.285 mmol). This material was used directly in the next step without further purification. LCMS: method 1, 1.46 min, MS: ES + 203.3.

And d, step d. To 1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline at rt]To a stirred solution of (0.08g, 0.253mmol) of the (E) -2' -keto TFA salt in THF (10ml) was added K₂CO₃(0.174g, 1.265 mmol). Cyanogen bromide (0.032g, 0.304mmol) was added to the reaction mixture at rt and the mixture was stirred at rt for 0.5 h. The resulting reaction mixture was filtered and excess THF was removed under reduced pressure. The resulting residue was purified by flash chromatography (30% EtOAc in hexane) to give 2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-1-nitrile (0.022g, 0.096 mmol). LCMS: method 2, 3.26 min, MS: ES + 228.4;¹H NMR(400 MHz，DMSO-d6)δppm：10.36(s，1H)，7.16-7.20(m，2H)，6.93-6.97 (m，1H)，6.88(d，J＝7.6Hz，1H)，3.67(d，J＝9.6Hz，1H)，3.51-3.55 (m，1H)，3.38-3.44(m，1H)，3.22(d，J＝9.6Hz，1H)，3.02(d，J＝ 16Hz，1H)，2.89(s，J＝15.6Hz，1H)，1.97-2.04(m，1H)，1.72-1.79 (m，1H)。

scheme 2

Reagents and conditions: a) ArB (OH)₂，Cs₂CO₃，Pd(PPh₃)₄1, 4-twoAlkane, water; b) TFA, DCM or HCl/EtOAc; c) CNBr, K₂CO₃THF or CNBr, NaHCO₃，EtOH

Example 42 '-oxo-7' -phenyl-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

Synthesis according to scheme 2

And a step a. To 7 '-bromo-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline at rt]Tert-butyl (1-carboxylate) (intermediate B; 0.25g, 0.655mmol) in 1, 4-bisAlkane: to a stirred solution in water (8: 2, 10ml) was added phenylboronic acid (0.16g, 1.311mmol) and Cs₂CO₃(0.427g, 1.311 mmol). The reaction mixture was degassed at rt for 20min, then Pd (PPh) was added₃)₄(0.075g, 0.065 mmol). The reaction mixture was heated at 80 ℃ for 8 hours. The resulting reaction mixture was cooled to rt, poured into water (30ml) and extracted with EtOAc (5 × 25 ml). The combined organic phases were separated and washed with brine (2 × 20 ml). Mixing the organic phaseSeparating with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (18% EtOAc in hexane) to give 2 '-oxo-7' -phenyl-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline]Tert-butyl 1-carboxylate (0.21g, 0.552 mmol). LCMS: method 1, 2.54 min, MS: ES +323.4 (M-56).

And (b) step (c). The title compound was synthesized following a procedure analogous to example 1, steps c-d, using the above intermediates. LCMS: method 2, 4.23 min, MS: ES + 304.3;¹H NMR(400MHz，DMSO-d₆)δppm 10.45(s，1H)，7.57-7.59(m，2 H)，7.45-7.49(m，2H)，7.35-7.39(m，1H)，7.25-7.28(m，2H)， 7.14(d，J＝1.6Hz，1H)，3.71(d，J＝9.6Hz，1H)，3.52-3.57(m，1H)， 3.40-3.46(m，1H)，3.26(d，J＝10.0Hz，1H)，3.07(d，J＝16.0Hz，1 H)，2.95(d，J＝16.0Hz，1H)，2.03-2.09(m，1H)，1.77-1.84(m，1 H)。

example 57 '- (5-isopropyl-2-methoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

Synthesis according to scheme 2

And a step a. To 7 '-bromo-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline under rt nitrogen]Tert-butyl (1-carboxylate) (intermediate B; 0.2mmol), (5-isopropyl-2-methoxyphenyl) boronic acid (0.2mmol) and Cs₂CO₃(0.6mmol, 3eq) in the 1, 4-bisPd (PPh) was added to a solution of an alkane (1ml) and water (0.2 ml)₃)₄(0.2 eq). The reaction mixture was stirred at 100 ℃ for 16 hours. The resulting mixture was concentrated under reduced pressure. The resulting residue was purified by preparative TLC (PE/EtOAc ═ 1: 1) to give 7' - (5-isopropyl-2-methoxyphenyl) -2' -oxo-2 ',4' -dihydro-1 ' H-spiro [ pyrrolidine ═-3,3' -quinolines]-1-carboxylic acid tert-butyl ester.

And b, step b. To a solution of 7'- (5-isopropyl-2-methoxyphenyl) -2' -oxo-2 ',4' -dihydro-1 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carboxylic acid tert-butyl ester in EtOAc (1ml) was added HCl/EtOAc (4M, 1 ml). The reaction mixture was stirred at rt for 2 hours. The resulting mixture was concentrated under reduced pressure. The residue 7' - (5-isopropyl-2-methoxyphenyl) -1' H-spiro [ pyrrolidine-3, 3' -quinolin ] -2' (4 ' H) -one was used directly in the next step without further purification.

And c, step (c). To 7' - (5-isopropyl-2-methoxyphenyl) -1' H-spiro [ pyrrolidine-3, 3' -quinoline]To a solution of-2 '(4' H) -ketone in EtOH (2ml) was added cyanogen bromide (0.2mmol) and NaHCO₃(0.6 mmol). The reaction mixture was stirred at rt for 16 h. The resulting mixture was concentrated under reduced pressure. The crude product was purified by preparative reverse phase HPLC (A: 0.078% CH)₃COONH₄The aqueous solution of (1), B: MeCN) to give 7'- (5-isopropyl-2-methoxyphenyl) -2' -oxo-2 ',4' -dihydro-1 'H-spiro [ pyrrolidine-3, 3' -quinoline]-1-nitrile (37.25mg, 99.2. mu. mol). LCMS: method 8, 3.353 min, MS: ES + 376.1.

The compounds in table 2 were synthesized using a procedure similar to that described in example 5.

TABLE 2

The compounds in tables 3.1 and 3.2 were synthesized using 4-bromo-2- (bromomethyl) -1-nitrobenzene using a procedure similar to that described for intermediate B/example 4.

TABLE 3.2

Example 461-cyano-N, N-dimethyl-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -carboxamide

And a step a. To 7 '-bromo-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline at rt in an autoclave]To a stirred solution of tert-butyl-1-carboxylate (intermediate B; 0.75g, 1.967mmol) in MeOH (20ml) were added sodium acetate (0.81g, 9.837mmol) and PdCl₂(dppf) DCM complex (0.32 g). The reaction mixture was heated at 120 ℃ for 4 days under a pressure of 30kg of carbon monoxide. The reaction mixture was cooled to rt, filtered through celite hyflow and washed with MeOH (5 × 30 ml). The filtrate was concentrated in vacuo and the resulting residue was purified by flash chromatography (38% EtOAc in hexanes) to give 2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]1- (tert-butyl) 7 '-methyl (1, 7' -dicarboxylate) (0.37g, 1.025 mmol). LCMS: method 1, 2.159 min, MS: ES +305.08 (M-56);¹H NMR(400MHz，DMSO-d₆)10.44(s， 1H)，7.50-7.56(m，2H)，7.34(d，J＝8.0Hz，1H)，3.83(s，3H)， 3.57(t，J＝9.6Hz，1H)，3.35-3.36(m，2H)，2.96-3.11(m，3H)， 1.91-2.01(m，1H)，1.68-1.74(m，1H)，1.39(d，J＝4.8Hz，9H)。

and b, step b. To 2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline at rt]To a stirred solution of 1- (tert-butyl) 7 '-methyl (0.35g, 0.969mmol) 1, 7' -dicarboxylate in THF: water (1: 1; 20ml) was added NaOH (0.077g, 1.939 mmol). The reaction mixture was stirred at rt for 16 h. The resulting reaction mixture was acidified with dilute HCl (30ml) and extracted with EtOAc (10 × 50 ml). The combined organic phases were separated over Na₂SO₄Drying, filtering and concentrating under reduced pressure to give 1- (tert-butyloxycarbonyl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-7' -carboxylic acid (0.26g, 0.749 mmol). LCMS: method 1, 1.929 min, MS: ES +291.1 (M-56);¹H NMR(400MHz，DMSO-d₆)δppm 12.92 (br s，1H)，10.43(s，1H)，7.48-7.57(m，2H)，7.30(d，J＝8.0Hz， 1H)，3.54-3.59(m，1H)，3.22-3.34(m，2H)，2.95-3.10(m，3H)， 1.99-2.01(m，1H)，1.68-1.73(m，1H)，1.38(d，J＝4.4Hz，9H)。

and c, step (c). To 1- (tert-butyloxycarbonyl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline at 0 deg.C]To a stirred solution of-7' -carboxylic acid (0.25g, 0.72mmol) in THF (10ml) were added HATU (0.41g, 1.08mmol) and DIPEA (0.185g, 1.44 mmol). The reaction mixture was stirred at 0 ℃ for 25 minutes. A solution of dimethylamine (2M in THF; 0.72 ml, 1.44mmol) was added to the reaction mixture at rt. The reaction mixture was stirred at rt for 16 h. The resulting reaction mixture was poured into water (50ml) and extracted with EtOAc (5 × 50 ml). The combined organic phases were separated and washed with saturated NaHCO₃The solution (25ml) was washed. Separating the organic phase with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (80% EtOAc in hexane) to give 7'- (dimethylcarbamoyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro- [ pyrrolidine-3, 3' -quinoline]Tert-butyl 1-carboxylate (0.26g, 0.696 mmol). LCMS: method 1, 1.909 min, MS: ES +318.18 (M-56);¹H NMR(400MHz， DMSO-d₆)δppm 10.36(s，1H)，7.23(d，J＝7.6Hz，1H)，6.95-6.97(m， 1H)，6.87(d，J＝1.2Hz，1H)，3.54-3.60(m，1H)，3.36-3.69(m， 1H)，3.21-3.28(m，1H)，3.07-3.11(m，1H)，2.88-3.03(m，8H)， 1.96-2.06(m，1H)，1.67-1.75(m，1H)，1.39(s，9H)。

and d, step d. To 7'- (dimethylcarbamoyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline at rt]To a stirred solution of tert-butyl-1-carboxylate (0.25g, 0.669mmol) in THF (3ml) was added TFA (0.25ml) dropwise. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under vacuum. The resulting crude material was washed with hexane and dried under vacuum to give N, N-dimethyl-2 ' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-7' -carboxamide TFA salt (0.21g, 0.542 mmol). LCMS: method 1, 1.289 min, MS: ES + 274.21;¹H NMR(400MHz，DMSO-d₆)δppm 10.6(s，1H)， 9.04(s，1H)，8.99(s，1H)，7.25(d，J＝7.6Hz，1H)，6.99(d，1.2Hz， 1H)，6.91(s，1H)，3.66(t，J＝4.8Hz，1H)，3.32-3.41(m，1H)， 3.16(t，J＝7.6Hz，2H)，3.00-3.06(m，1H)，2.91-2.97(m，6H)， 2.01-2.07(m，1H)，1.82-1.87(m，1H)。

and e, step e. To N, N-dimethyl-2 ' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline at 0 DEG C]To a stirred solution of (E) -7' -carboxamide TFA salt (0.2g, 0.516mmol) in THF (12ml) was added K₂CO₃(0.356g, 2.584 mmol). The reaction mixture was stirred at rt for 2 hours. The resulting reaction mixture was filtered, washed with THF (30ml) and the filtrate was concentrated in vacuo. The resulting residue was purified by flash chromatography (column packed in hexanes; gradient of EtOAc was gradually increased to 100%) to give 1-cyano-N, N-dimethyl-2 ' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-7' -carboxamide (0.13g, 0.435 mmol). LCMS: method 2, 2.833 min, MS: ES + 299.21;¹H NMR(400MHz，DMSO-d₆)δ ppm 10.46(s，1H)，7.24(d，J＝7.6Hz，1H)，6.97(dd，J＝1.6Hz，7.6 Hz，1H)，6.89(s，1H)，3.69(d，J＝9.6Hz，1H)，3.51-3.56(m，1H)， 3.39-3.45(m，2H)，3.24(d，J＝9.6Hz，1H)，3.05(d，J＝16.0Hz，1H)， 2.91-2.97(m，6H)，2.00-2.07(m，1H)，1.74-1.81(m，1H)。

example 471-cyano-N-methyl-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -carboxamide

Steps a-e. The title compound was synthesized following a procedure similar to that of example 46, using methylamine (2M in THF) in step c. LCMS: method 2RT 2.493 min, MS: ES + 285.27;¹H NMR(400MHz，DMSO-d6)δppm：10.47(s，1H)， 8.35(d，J＝4.8Hz，1H)，7.35-7.39(m，2H)，7.26(d，J＝8Hz，1H)， 3.68(d，J＝9.6Hz，1H)，3.49-3.55(m，1H)，3.38-3.44(m，1H)， 3.22(d，J＝9.6Hz，1H)，3.06(d，J＝16.0Hz，1H)，2.95(d，J＝16.4Hz， 1H)，2.75(d，J＝4.4Hz，3H)，1.97-2.03(m，1H)，1.74-1.78(m， 1H)。

example 132(R) -7'- (5-methyl-1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

And a step a. To a solution of 4-bromo-2-nitrobenzaldehyde (CAS number 5551-12-2; 10.000g, 43.478mmol) in MeOH (120ml) at rt was added isopropylidene malonate (CAS number 2033-24-1; 6.260g, 43.478mmol), diethyl 1, 4-dihydro-2, 6-dimethyl-3, 5-pyridinedicarboxylate (CAS number 1149-23-1; 11.000g, 43.478mmol) and L-proline (CAS number 147-85-3; 0.99g, 8.690 mmol). The reaction mixture was stirred at rt for 16 h. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was purified by flash chromatography (15% EtOAc in hexane) to give 5- (4-bromo-2-nitrobenzyl) -2, 2-dimethyl-1, 3-di-methylAlkane-4, 6-dione (14.500g, 40.503 mmol). LCMS: method 1, 3.269 min, MS: ES +358.0, 359.0

And b, step b. In the rt direction to 5- (4-bromo-2-nitrobenzyl) -2, 2-dimethyl-1, 3-bisTo a solution of alkane-4, 6-dione (14.00g, 39.106mmol) in MeOH (120ml) was added N, N-dimethylmethylammonium iodide (CAS number 33797-51-2; 18.08g, 97.297 mmol). The reaction mixture was heated to 70 ℃ for 16 hours. The resulting reaction mixture was cooled to rt and distilled under reduced pressure. The resulting mixture was dissolved in diethyl ether (500ml) and saturated NaHCO was used₃The solution (3 × 100ml) was washed. The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (15% EtOAc in hexane) to give ethyl 2- (4-bromo-2-nitrobenzyl) acrylate (10.0)0g, 31.847 mmol). This material was used in the next step without further purification.

And c, step (c). To a solution of ethyl 2- (4-bromo-2-nitrobenzyl) acrylate (10.00g, 31.847mmol) in MeCN (80ml) was added a solution of N- (methoxymethyl) -N- (trimethylsilylmethyl) aniline (CAS number 93102-05-7; 9.829g, 41.471mmol) in MeCN (20ml) at 0 ℃. AgF (4.444g, 35.030mmol) was added to the reaction mixture in portions at 0 ℃. The reaction mixture was stirred at rt for 16 h. The resulting mixture was filtered through celite and washed with EtOAc (100 ml). The filtrate was diluted with water (100ml) and extracted with EtOAc (10 × 100 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (11% EtOAc in n-hexane) to give methyl 1-benzyl-3- (4-bromo-2-nitrobenzyl) pyrrolidine-3-carboxylate (11.500g, 26.558 mmol). LCMS: method 1, 1.904, MS: ES +433.1, 435.1

And d, step d. To a solution of methyl 1-benzyl-3- (4-bromo-2-nitrobenzyl) pyrrolidine-3-carboxylate (11.50 g, 26.558mmol) in THF (100ml) at rt was added NH₄A solution of Cl (14.20g, 265.47 mmol) in water (100ml) was followed by the addition of Fe powder (14.81g, 265.41 mmol). The reaction mixture was heated to 80 ℃ for 16 hours. The resulting mixture was cooled to rt, filtered through celite and washed with EtOAc (5 × 100 ml). The resulting filtrate was diluted with water (200ml) and extracted in EtOAc (10 × 100 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (80-100% EtOAc in hexane) to afford 1-benzyl-7 ' -bromo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-2' -ketopyridine (7.140g, 19.245 mmol). LCMS: method 1, 1.781, MS: ES +371.1, 373.1

And e, step e. To 1-benzyl-7 ' -bromo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline at rt]-2' -ketopyridine (0.300g, 0.801mmol) and 5-methyl-1 h-indazol-4-ylboronic acid (CAS number 1245816-10-7; 0.214g, 1.216mmol) in 1, 4-bisCs was added to a stirred solution of alkane and water (4: 1, 12ml)₂CO₃(0.528g, 1.624 mmol). The reaction mixture was degassed for 15 minutes, then Pd (PPh) was added at rt₃)₄(0.046g, 0.039 mmol). The reaction mixture was heated at 80 ℃ for 16 hours. The resulting mixture was cooled to rt, diluted with water (20ml) and extracted with EtOAc (5 × 50 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (40-80% EtOAc in hexanes) to give 1-benzyl-7 ' - (5-methyl-1H-indazol-4-yl) -1',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-2' -ketone (0.200g, 0.473 mmol). LCMS: method 1, 1.783 min, MS: ES + 423.52.

And f, step d. To 1-benzyl-7 ' - (5-methyl-1H-indazol-4-yl) -1',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline at 0 deg.C]Addition of K to a solution of (0.180g, 0.426mmol) of (E) -2' -one in THF (10ml)₂CO₃(0.058g, 0.420mmol) and CNBr (0.045g, 0.426 mmol). The reaction mixture was warmed to rt and stirred at rt for 6 hours. The resulting mixture was poured into water (50ml) and extracted with EtOAc (5 × 50 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (3% MeOH in DCM) and further purified by preparative TLC using (3% MeOH in DCM) to give the racemate of the desired compound (0.037g, 0.104 mmol). LCMS: method 1, 1.776 min, MS: the ES +358.40 is set to a value of ES +358.40,¹H NMR(400MHz，DMSO-d6) δppm：13.05(s，1H)，10.40(s，1H)，7.64(s，1H)，7.43-7.45(m， 1H)，7.28-7.34(m，2H)，7.00-7.02(m，1H)，6.95(s，1H)，3.75(d， J＝9.6Hz，1H)，3.56-3.58(m，1H)，3.45-3.47(m，1H)，3.30(d， J＝9.6Hz，1H)，3.09-3.13(m，1H)，2.98-3.02(m，1H)，2.28(s， 3H)，2.09-2.14(m，1H)，1.82-1.87(m，1H)。

on a Chiralpak IB 250X20.0 mm, 5micron column, liquid CO was used₂25% IPA in (13 min), flow rate of 70.0 ml/min and 100 bar ABPR, enantiomers were separated by chiral SFC purification to yieldThe title compound. Absolute stereochemistry was specified by a method similar to example 63. LCMS: method 1RT 1.888 min, MS: ES + 358.32; chiral HPLC: column Chiralpak IB 250 × 4.6mm, 5 microns, flow 1 ml/min, 6.82 min, 40% IPA in n-hexane solution RT 9.26 min;¹H NMR(400MHz， DMSO-d6)δppm：13.05(s，1H)，10.40(s，1H)，7.64(s，1H)，7.43 -7.45(m，1H)，7.28-7.34(m，2H)，7.00-7.02(m，1H)，6.95(s，1 H)，3.75(d，J＝9.6Hz，1H)，3.56-3.58(m，1H)，3.45-3.47(m，1 H)，3.30(d，J＝9.6Hz，1H)，3.09-3.13(m，1H)，2.98-3.02(m，1 H)，2.28(s，3H)，2.09-2.14(m，1H)，1.82-1.87(m，1H)。

example 1337 '- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

This compound was prepared using a method analogous to steps a-f of example 132, using indazole-4-boronic acid (CAS number 1023595-17-6) in step e. LCMS: method 3 RT 3.815 min, MS: ES + 343.99;¹H NMR(400MHz，DMSO-d6)δppm：13.24 (s，1H)，10.44(s，1H)，8.18(s，1H)，7.54(d，J＝8.4Hz，1H)，7.40 -7.44(m，1H)，7.31-7.36(m，3H)，7.20(d，J＝7.2Hz，1H)，3.73 (d，J＝9.6Hz，1H)，3.53-3.58(m，1H)，3.41-3.50(m，1H)，3.28 (d，J＝9.6Hz，1H)，3.08-3.12(m，1H)，2.88-2.97(m，1H)，2.06 -2.12(m，1H)，1.79-1.86(m，1H)。

example 1346 '- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile

This compound was prepared in a similar manner to example 133 using 5-bromo-2-nitrobenzaldehyde (CAS number 20357-20-4) in the first step. LCMS: method 3RT 3.517 min, MS: ES + 344.06;¹H NMR(400MHz，DMSO-d6)δppm：13.19(s，1H)， 10.50(s，1H)，8.23(s，1H)，7.56-7.59(m，2H)，7.50(d，J＝8.4Hz， 1H)，7.40(t，J＝8.4Hz，1H)，7.20(d，J＝6.4Hz，1H)，7.04(d，J＝ 8.0Hz，1H)，3.73(d，J＝9.6Hz，1H)，3.54-3.62(m，1H)，3.39-3.47 (m，1H)，3.30(d，J＝9.6Hz，1H)，3.02-3.17(m，2H)，2.03-2.09 (m，1H)，1.78-1.88(m，1H)。

scheme 3

Reagents and conditions: a) cs₂CO₃，DMF，60℃；b)Fe，NH₄Cl, THF, water, 60 ℃; c) TFA, DCM, 0 ℃; d) CNBr, K₂CO₃，THF，0℃

Example 482-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

Synthesis according to scheme 3

And a step a. To a stirred solution of 2-chloro-3-nitropyridine (CAS number 5470-18-8; 0.5g, 3.154 mmol) in DMF (10ml) at rt was added 3-hydroxypyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (intermediate C; 0.62g, 2.524mmol) and Cs₂CO₃(3.08g, 9.463 mmol). The reaction mixture was stirred at 60 ℃ for 16 hours. The resulting reaction mixture was poured into ice cold water (100ml) and extracted with EtOAc (2 × 70 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography using neutral alumina (25% EtOAc in hexane) to afford 3- ((3-nitropyridin-2-yl) oxy) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-Methyl ester (0.12g, 0.327 mmol). LCMS: method 1, 2.20 min, MS: ES + 368.5;¹H NMR(400MHz，DMSO-d6) δppm：8.51(d，J＝8.0Hz，1H)，8.44(dd，J＝1.6Hz，4.8Hz，1H)， 7.30-7.34(m，1H)，3.97-4.04(m，1H)，3.68-3.71(m，1H)，3.63(s， 3H)，3.44-3.48(m，2H)，2.41-2.45(m，2H)，1.38(s，9H)。

and b, step b. To a stirred solution of 3- ((3-nitropyridin-2-yl) oxy) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (0.1g, 0.272mmol) in THF: water (1: 1; 10ml) at rt was added iron powder (0.15g, 2.724mmol) and NH₄Cl (0.15g, 2.724 mmol). The reaction mixture was stirred at 60 ℃ for 16 hours. The resulting reaction mixture was filtered through a celite bed. The filtrate was poured into water (50ml) and extracted with EtOAc (2 × 30 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography using neutral alumina (5% MeOH in DCM) to give 2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carboxylic acid tert-butyl ester (0.065g, 0.213 mmol). LCMS: method 1, 1.95 min, MS: ES + 306.3;¹H NMR(400 MHz，DMSO-d6)δppm：11.09(s，1H)，7.85(d，J＝4.4Hz，1H)，7.29 (dd，J＝1.6Hz，7.6Hz，1H)，7.09(dd，J＝4.8Hz，7.6Hz，1H)，3.68 -3.76(m，1H)，3.50-3.57(m，2H)，3.39-3.45(m，1H)，2.32-2.40 (m，1H)，2.14-2.17(m，1H)，1.39(s，9H)。

and c, step (c). To 2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] at 0 deg.C][1，4]Oxazine-3, 3' -pyrrolidines]To a stirred solution of tert-butyl (1' -carboxylate) (0.06g, 0.197mmol) in DCM (5ml) was added TFA (0.3 ml). The reaction mixture was stirred at 0 ℃ for 1 hour. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was azeotropically distilled using DCM (3 × 5ml) and triturated with hexane (2 × 3ml) to give spiro [ pyrido [2,3-b ] s][1，4]Oxazine-3, 3' -pyrrolidines]-2(1H) -one TFA salt (0.08g, quant.). This material was used directly in the next step without further purification. LCMS: method 1, 0.44 min, MS: ES + 206.2;¹H NMR (400MHz，DMSO-d6)δppm：11.29(s，1H)，9.48(br，s，1H)，7.80 (d，J＝4.0Hz，1H)，7.33(d，J＝7.2Hz，1H)，7.13(t，J＝6.0Hz，1H)， 3.77-3.81(m，1H)，3.63-3.66(m，1H)，3.39-3.45(m，2H)，2.31 -2.36(m，2H)。

and d, step d. Spiro [ pyrido [2,3-b ] at 0 deg.C][1，4]Oxazine-3, 3' -pyrrolidines]To a stirred solution of (E) -2(1H) -one TFA salt (0.08g, 0.25mmol) in THF (5ml) was added K₂CO₃(0.17g, 1.253 mmol). The mixture was stirred at 0 ℃ for 5 minutes. Cyanogen bromide (0.032g, 0.301mmol) was added to the reaction mixture at 0 ℃. The reaction mixture was stirred at 0 ℃ for 15 minutes. The resulting reaction mixture was filtered and excess THF was distilled under reduced pressure. The resulting residue was purified by flash chromatography using neutral alumina (5% MeOH in DCM) to give 2-oxo-1, 2-dihydrospiro- [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -nitrile (0.04g, 0.174 mmol). LCMS: method 3, 2.40 min, MS: ES + 231.1;¹H NMR(400MHz，DMSO-d6)δppm：11.18(s，1H)，7.87(d，J＝8.0 Hz，1H)，7.30(d，J＝6.8Hz，1H)，7.10(dd，J＝4.8Hz，7.2Hz，1H)，3.86(d，J＝11.2Hz，1H)，3.71(d，J＝11.6Hz，1H)，3.57-3.67(m，2 H)，2.35-2.43(m，1H)，2.21-2.25(m，1H)。

scheme 4

Reagents and conditions: a) pd (PPh)₃)₄，Cs₂CO₃1, 4-twoAlkane, water; b) TFA, DCM; c) cyanogen bromide, K₂CO₃，THF

Example 492-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

And a step a. To 7-bromo-2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] at rt][1，4]Oxazine-3, 3' -pyrrolidines]Tert-butyl (1' -carboxylate) (intermediate D; 0.25g, 0.651mmol) in 1, 4-bisCs was added to a stirred solution of alkane and water (4: 1; 10ml)₂CO₃(0.423g, 1.301 mmol). The reaction mixture was degassed for 20 minutes, then Pd (PPh) was added at rt₃)₄(0.075g, 0.065mmol) and phenylboronic acid (0.158g, 1.301 mmol). The reaction mixture was heated at 90 ℃ for 18 hours. The resulting reaction mixture was cooled to rt, poured into water (20ml) and extracted with EtOAc (3 × 30 ml). The combined organic phases were separated and washed with brine (20 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography (30% EtOAc in hexane) to give 2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carboxylic acid tert-butyl ester(0.22g, 0.783 mmol). LCMS: method 1, 2.328 min, MS: ES + 382.3;¹H NMR(400MHz， DMSO-d₆)δppm 11.19(s，1H)，8.16(s，1H)，7.62(d，J＝7.2Hz，2H)， 7.50(t，J＝7.6Hz，3H)，7.41(d，J＝7.2Hz，1H)，3.72-3.80(m，1H)， 3.54-3.62(m，2H)，3.44-3.51(m，1H)，2.18-2.24(m，1H)，2，.33 -2.41(m，1H)，1.42(d，J＝10.0Hz，9H)。

and b, step b. To 2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ] at rt][1，4]Oxazine-3, 3' -pyrrolidines]To a stirred solution of tert-butyl (1' -carboxylate) (0.21g, 0.551mmol) in DCM (10ml) was added TFA (1.05 ml). The reaction mixture was stirred at rt for 5 hours. The resulting reaction mixture was concentrated under vacuum. The resulting residue was azeotropically distilled with DCM (2 × 20 ml). The resulting material was washed with diethyl ether (2 × 20ml) and dried under vacuum to give 7-phenylspiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-2(1H) -keto TFA salt (0.21g, 0.531 mmol). LCMS: method 1, 1.578 min, MS: ES + 282.18.

And c, step (c). To 7-phenylspiro [ pyrido [2,3-b ] at 0 deg.C][1，4]Oxazine-3, 3' -pyrrolidines]To a stirred solution of (E) -2(1H) -one TFA salt (0.2g, 0.506mmol) in THF (10ml) was added K₂CO₃(0.208, 1.517 mmol). The reaction mixture was stirred at 0 ℃ for 10 minutes. Cyanogen bromide (0.063g, 0.607mmol) at 0 ℃. The reaction mixture was stirred at rt for 30 min. The resulting reaction mixture was poured into water (10ml) and extracted with EtOAc (3 × 30 ml). The combined organic phases were separated and washed with brine (20 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (1.4% MeOH in DCM) to give 2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b][1，4]oxazine-3, 3' -pyrrolidines]-1' -nitrile (0.085g, 0.278 mmol). LCMS: method 1, 1.855 min, MS: ES + 307.37;¹H NMR(400MHz，DMSO-d₆)δppm 11.28(s，1H)， 8.17(d，J＝2.0Hz，1H)，7.62(d，J＝1.2Hz，2H)，7.50(t，J＝7.2Hz， 3H)，7.41(J＝7.6Hz，1H)，3.88(d，J＝11.6Hz，1H)，3.77(dd，J＝1.2 Hz，11.2Hz，1H)，3.60-3.72(m，2H)，2.38-2.44(m，1H)，2.27- 2.32(m，1H)。

example 542-oxo-6-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

This compound was prepared by an analogous method to example 49 using intermediate G and 2, 6-dichloro-3-nitropyridine. LCMS: method 2RT 3.624 min, MS: ES + 307.21;¹H NMR(400MHz，DMSO-d6)δppm：11.25(s，1H)，7.97(d，J＝7.6 Hz，2H)，7.69(d，J＝8Hz，1H)，7.36-7.48(m，4H)，3.89(d，J＝11.2 Hz，1H)，3.78(d，J＝11.2Hz，1H)，3.64-3.68(m，2H)，2.39-2.41 (m，1H)，2.30-2.32(m，1H)。

example 1396- (1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

This compound was prepared by an analogous method to example 135 using intermediate G and 2, 6-dichloro-3-nitro-pyridine. LCMS: method 3 RT 2.947 min, MS ES + 347.05;¹H NMR(400MHz，DMSO-d6)δppm：13.20(s，1H)，11.31(s，1H)， 8.59(s，1H)，7.80(d，J＝8.0Hz，1H)，7.63(d，J＝7.2Hz，1H)，7.58 (d，J＝8.4Hz，1H)，7.41-7.44(m，2H)，3.88-3.91(m，1H)，3.80 -3.83(m，1H)，3.67-3.71(m，2H)，2.38-2.43(m，1H)，2.32-2.36 (m，1H)。

example 1407- (1- (2-methoxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，41Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

This compound was prepared in analogy to intermediate F/example 137 using 2-bromoethyl methyl ether (CAS number 6482-24-2). LCMS: method 3 RT 3.072 min, MS: ES + 405.15;¹H NMR(400MHz，DMSO-d6)δppm：11.26(s，1H)， 8.23(d，J＝2.0Hz，1H)，8.19(s，1H)，7.73(d，J＝8.8Hz，1H)，7.64 (d，J＝2.0Hz，1H)，7.48(t，J＝8.4Hz，1H)，7.26(d，J＝6.8Hz，1H)， 4.61-4.63(m，2H)，4.00-4.03(m，1H)，3.77-3.90(m，3H)，3.61 -3.71(m，2H)，3.20(s，3H)，2.41-2.44(m，1H)，2.32-2.36(m， 1H)。

example 1411' -cyano-N- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2.3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-6-carboxamides

And a step a. To 6-bromo-2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] at rt][1，4]Oxazine-3, 3' -pyrrolidines]Tert-butyl (1' -carboxylate) (intermediate G; 0.300G, 0.783mmol) to a solution in anhydrous MeOH (5ml) was added NaOAc (0.322G, 3.912mmol) and prepared in an autoclave. The reaction mixture was degassed for 30 minutes, then PdCl was added at rt₂(dppf) DCM complex (0.046g, 0.039mmol) and 25kg/cm²H₂Pressure is applied in the autoclave. The reaction mixture was heated to 100 ℃ for 48 hours. The resulting reaction mixture was combined with another batch prepared by the same method on the same scale. The resulting reaction mixture was cooled to rt and filtered through celite. The resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by flash chromatography (1.7% MeOH in DCM) to give 2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]1'- (tert-butyl) 6-methyl 1', 6-dicarboxylate (0.400g, 1.102 mmol). LCMS: method 1, 1.658 min, MS: ES +364.58[ M +1 ]]。

And b, step b. To 2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] at rt][1，4]Oxazine-3, 3' -pyrrolidines]To a solution of 1'- (tert-butyl) 6-methyl (0.400g, 1.102mmol) of 1', 6-dicarboxylate in THF: water (1: 1, 5ml) was added NaOH (0.088g, 2.203 mmol).The reaction mixture was stirred at rt for 16 h. The resulting reaction mixture was poured into water (50ml) and acidified using a saturated citric acid solution. The resulting mixture was extracted in EtOAc (3 × 50 ml). The combined organic phases were washed with Na₂SO₄Drying, filtering and concentrating under reduced pressure to obtain 1' - (tert-butyloxycarbonyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]6-Carboxylic acid (0.350g, 1.003 mmol). This material was used directly in the next step without further purification. LCMS: method 1, 1.507 minutes. MS: ES +350.60[ M +1 ]]

And c, step (c). To 1' - (tert-butyloxycarbonyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] at rt][1，4] Oxazine-3, 3' -pyrrolidines]To a solution of-6-carboxylic acid (0.330g, 0.945mmol) in THF (5ml) were added HATU (0.538g, 1.418mmol) and DIPEA (0.244g, 1.890 mmol). The reaction mixture was stirred at rt for 30 min. 4-fluoroaniline (CAS number 371-40-4; 0.126g, 1.134mmol) was added to the reaction mixture at rt. The reaction mixture was stirred at rt for 3 hours. The resulting mixture was poured into water (50ml) and extracted with EtOAc (3 × 50 ml). The combined organic phases were dried over Na₂SO₄Filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (2.2% MeOH in DCM) to give 6- ((4-fluorophenyl) carbamoyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -carboxylic acid tert-butyl ester (0.250g, 0.565 mmol). LCMS: method 1, 2.059 min, MS: ES +443.70[ M +1 ]]

And d, step d. To 6- ((4-fluorophenyl) carbamoyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] at 0 deg.C][1，4]Oxazine-3, 3'-pyrrolidines]To a solution of tert-butyl (1' -carboxylate) (0.250g, 0.565mmol) in DCM (10ml) was added TFA (1.25ml, 5V). The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was co-distilled using MTBE (3 × 5 ml). The resulting residue was triturated with MTBE (2 × 5 ml). Drying the resulting residue under high vacuum to give N- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-6-carboxamide TFA salt (0.250 g). This material was used directly in the next step without further purification. LCMS: method 1, 1.406 min, MS: ES +343.50[ M +1 ]]

And e, step e. To N- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] at rt][1，4] Oxazine-3, 3' -pyrrolidines]To a solution of-6-carboxamide TFA salt (0.250g, 0.548mmol) in THF (5ml) was added K₂CO₃(0.234g, 1.695 mmol). The reaction mixture was stirred at rt for 15 minutes. The reaction mixture was cooled to 0 ℃ and treated with CNBr (0.058g, 0.547 mmol). The reaction mixture was stirred for 1 hour, then poured into water (50 ml). The resulting precipitate was collected by filtration and dried under high vacuum to give the title compound (0.150g, 0.408 mmol). LCMS: method 3 RT 3.558 min, MS ES-366.05;¹H NMR(400 MHz，DMSO-d6)δppm：11.52(s，1H)，10.38(s，1H)，7.85-7.86(m， 3H)，7.43-7.45(m，1H)，7.13-7.18(m，2H)，3.90-3.93(m，1H)， 3.80-3.83(m，1H)，3.62-3.68(m，2H)，2.38-2.43(m，1H)，2.29 -2.33(m，1H)。

example 1422-oxo-6- (piperidine-1-carbonyl) -1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4] Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

This compound was prepared by an analogous method to example 141, using piperidine in step c. LCMS: method 2RT 2.859 min, MS ES + 342.42;¹H NMR(400MHz， DMSO-d6)δppm：11.34(s，1H)，7.36(d，J＝8.0Hz，1H)，7.26(d， J＝7.6Hz，1H)，3.84-3.87(m，1H)，3.75-3.77(m，1H)，3.50-3.70 (m，6H)，2.36-2.42(m，1H)，2.26-2.33(m，1H)，1.45-1.61(m， 6H)。

example 1431' -cyano-2-oxo-N-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4] Oxazine-3, 3' -pyrrolidines]-6-carboxamides

This compound was prepared by an analogous method to example 141, using aniline in step c. LCMS: method 3 RT 3.337 min, MH + ES-348.05;¹H NMR(400 MHz，DMSO-d6)δppm：11.55(s，1H)，10.29(s，1H)，7.85-7.90(m， 3H)，7.47(d，J＝8.0Hz，1H)，7.34(t，J＝8.0Hz，2H)，7.10(t，J＝ 7.2Hz，1H)，3.93-3.96(m，1H)，3.83-3.86(m，1H)，3.63-3.73(m， 2H)，2.41-2.45(m，1H)，2.33-2.36(m，1H)。

example 1441' -cyano-N- (2-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-6-carboxamides

This compound was prepared by an analogous method to example 141 using 2-fluoroaniline in step c. L isCMS: method 3, RT 3.392 min, MS ES-366.05;¹H NMR (400MHz，DMSO-d6)δppm：11.58(s，1H)，10.07(s，1H)，7.93-7.95 (m，1H)，7.89(d，J＝8.0Hz，1H)，7.48(d，J＝7.6Hz，2H)，7.29- 7.32(m，1H)，7.22-7.24(m，2H)，3.91-3.93(m，1H)，3.85-3.88 (m，1H)，3.65-3.71(m，2H)，2.41-2.45(m，1H)，2.33-2.36(m， 1H)。

scheme 5

Reagents and conditions: a) k₂CO₃Toluene; b) fe, NH₄Cl, THF, water; c) CNBr, K₂CO₃，THF

Example 552-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

Synthesis according to scheme 5

And a step a. To a stirred solution of 3-amino-1-benzylpyrrolidine-3-carboxylic acid methyl ester (intermediate A; 0.3g, 1.280mmol) and 2-fluoro-3-nitropyridine (CAS number 1480-87-1; 0.236g, 1.665 mmol) in toluene (15ml) at rt was added K₂CO₃(0.265g, 1.921 mmol). The reaction mixture was heated at 120 ℃ for 16 hours. The reaction mixture was cooled to rt, poured into water (50ml) and extracted with EtOAc (3 × 50 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (10-15% EtOAc in hexane) to give methyl 1-benzyl-3- ((3-nitropyridin-2-yl) amino) pyrrolidine-3-carboxylate (0.17g, 0.477 mmol). LCMS: method 1, 1.82 min, MS: ES + 357.3;¹H NMR(400MHz，DMSO-d₆)δppm：8.45-8.47(m，1H)，8.37-8.40 (m，1H)，7.90(br，s，1H)，7.33(d，J＝4.4Hz，4H)，7.23-7.28(m， 1H)，6.83-6.87(m，1H)，6.74-6.77(m，1H)，3.60-3.70(m，2H)， 3.54(s，3H)，3.07(d，J＝10.4Hz，1H)，2.90(d，J＝10.0Hz，1H)， 2.83-2.89(m，1H)，2.55-2.62(m，1H)，2.12-2.16(m，1H)。

and b, step b. To a stirred solution of methyl 1-benzyl-3- ((3-nitropyridin-2-yl) amino) pyrrolidine-3-carboxylate (0.19g, 0.534mmol) in THF (5ml) at rt was added a solution of ammonium chloride (0.285 g, 5.331mmol) in water (5 ml). Iron powder (0.29g, 5.337mmol) was added at rt and the reaction mixture was heated at 60 ℃ for 2 h. The mixture was cooled to rt, poured into water (50ml) and extracted with EtOAc (2 × 50 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (2% MeOH in DCM) to give 1' -benzyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ]]Pyrazine-3, 3' -pyrrolidines]-2-one (0.08g, 0.272 mmol). LCMS: method 1, 1.39 min, MS: ES + 295.5;¹H NMR(400MHz，DMSO-d₆)δppm：10.46(s，1H)，7.65(dd， J＝1.6Hz，5.2Hz，1H)，7.30-7.31(m，4H)，7.21-7.24(m，1H)， 7.01(s，1H)，6.97(d，J＝6.8Hz，1H)，6.60-6.63(m，1H)，3.58(s， 2H)，2.84-2.87(m，1H)，2.63(q，J＝9.6Hz，2H)，2.38-2.43(m， 2H)，1.81-1.84(m，1H)。

and c, step (c). To 1' -benzyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] at 0 deg.C]Pyrazine-3, 3' -pyrrolidines]To a stirred solution of (0.07g, 0.238mmol) of (E) -2-one in THF (10ml) was added K₂CO₃(0.066g, 0.476 mmol). The reaction was stirred at 0 ℃ for 5 minutes. Cyanogen bromide (0.025g, 0.238mmol) was added to the reaction mixture at 0 ℃ and the reaction mixture was stirred at rt for 16 h. The resulting mixture was poured into water (50ml) and extracted with EtOAc (3 × 50 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (2% MeOH in DCM) to give 2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ]]Pyrazine-3, 3' -pyrrolidines]-1' -nitrile (0.03g, 0.131 mmol). LCMS: method 3, 2.44 min, MS: ES + 230.0;¹H NMR(400MHz， DMSO-d₆)δppm：10.71(s，1H)，7.70(dd，J＝1.6Hz，5.2Hz，1H)， 7.39(s，1H)，7.02(d，J＝7.6Hz，1H)，6.65-6.69(m，1H)，3.78(d， J＝10.0Hz，1H)，3.65-3.71(m，1H)，3.46(q，J＝8.0Hz，1H)，3.26 -3.29(m，1H)，2.27-2.33(m，1H)，1.90-1.97(m，1H)。

example 562-oxo-6- (trifluoromethyl) -1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

This compound was prepared using a procedure similar to that of example 55, using 2-chloro-3-nitro-6-trifluoromethylpyridine in step a (CAS number 117519-08-1). LCMS: method 2, 3.439 min, MS: ES + 298.18;¹H NMR(400MHz，DMSO-d₆)δppm： 11.08(s，1H)，8.13(s，1H)，7.10-7.15(m，2H)，3.80(d，J＝10.4Hz， 1H)，3.67-3.73(m，1H)，3.46-3.52(m，1H)，3.37(d，J＝10.0Hz， 1H)，2.33-2.42(m，1H)，1.96-2.02(m，1H)。

scheme 6

Reagents and conditions: a) k₂CO₃Toluene; b) fe, NH₄Cl, THF, water; c) ArB (OH)₂，Pd(PPh₃)₄，Cs₂CO₃1, 4-twoAlkane, water; d) CNBr, K₂CO₃， THF

Example 572-oxo-7-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

Prepared according to scheme 6

And a step a-b. These steps were carried out using procedures analogous to steps a and b of example 55, using 5-bromo-2-fluoro-3-nitropyridine in step a (CAS number 886372-98-1).

And c, step (c). This step was carried out using a procedure analogous to that of step a of example 4, using phenylboronic acid.

And d, step d. The title compound was formed by using a procedure analogous to step c of example 55. LCMS: method 3, 3.616 min, MS: ES + 306.0;¹H NMR(400MHz， DMSO-d₆)δppm：10.83(s，1H)，8.03(s，1H)，7.37-7.57(m，5H)， 7.29-7.37(m，2H)，3.80-3.82(m，1H)，3.69-3.70(m，1H)，3.48 -3.49(m，1H)，3.38-3.41(m，1H)，2.33-2.35(m，1H)，1.99-2.00 (m，1H)。

the compounds in table 5 were prepared in a similar manner to example 57.

Example 613-oxo-3, 4-dihydro-1H-spiro [ pyrido [2,3-b ] pyrazine-2, 3 '-pyrrolidine ] -1' -carbonitrile

Synthesized using a similar procedure as described in example 57, using 3-fluoro-2-nitropyridine in step a. LCMS: method 2RT 2.247 min, MS: ES + 230.25;¹H NMR (400MHz，DMSO-d6)δppm：11.02(s，1H)，7.65(d，J＝5.2Hz，1H)， 7.06(d，J＝7.6Hz，1H)，6.84-6.87(m，1H)，6.76(s，1H)，3.78(d， J＝10.4Hz，1H)，3.61-3.67(m，1H)，3.47-3.53(m，1H)，3.29(d， J＝10.0Hz，1H)，2.21-2.36(m，1H)，1.84-1.90(m，1H)。

example 1472-oxo-6-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

This compound was prepared using a procedure similar to that of example 57 using 6-bromo-2-chloro-3-nitropyridine in step a (CAS number 1430341-84-6). LCMS: method 2, MS: 3.675 minutes, ES + 306.32;¹H NMR(400MHz，DMSO-d₆)δppm：10.81(s，1 H)，7.92-7.94(m，2H)，7.53(s，1H)，7.40-7.44(m，2H)，7.32-7.36 (m，1H)，7.26(d，J＝8.0Hz，1H)，7.10(d，J＝7.6Hz，1H)，3.80(d， J＝10.4Hz，1H)，3.68-3.73(m，1H)，3.45-3.51(m，1H)，3.39(d， J＝10.4Hz，1H)，2.31-2.37(m，1H)，1.96-2.03(m，1H)。

example 626-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

And a step a. To 6-oxo-2, 7-diazaspiro [4.4] at 0 deg.C]To a solution of tert-butyl nonane-2-carboxylate (CAS number 1194376-44-7; 0.2g, 0.83mmol) in DCM (15ml) was added TFA (0.19ml, 2.4 mmol). The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was azeotropically distilled using DCM (2 × 5 ml). The resulting residue was triturated with diethyl ether (2X5ml) to give 2, 7-diazaspiro [4.4]]Nonan-1-one TFA salt (0.31g, quantitative). This material was used directly in the next step without further purification. LCMS: method 4, 2.26 min, MS: ES + 140.9;¹H NMR(400MHz， DMSO-d₆)δppm 5.01(br s，1H)，3.34-3.41(m，2H)，3.20-3.32(m， 4H)，3.12-3.18(m，1H)，2.04-2.14(m，2H)，1.94-2.02(m，2H)。

and b, step b. To 2, 7-diazaspiro [4.4] at rt]To a solution of nonan-1-one TFA salt (0.30g, 1.10mmol) in DMF (10ml) was added K₂CO₃(0.48g, 3.50 mmol). The reaction mixture was stirred at rt for 10 min. The reaction mixture was cooled to 0 ℃. Cyanogen bromide (0.15g, 1.40mmol) was added to the reaction mixture at 0 ℃. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was poured into water (40ml) and purified with 25% IPA: CHCl₃The mixture was extracted (5 × 40 ml). The combined organic phases were collected over Na₂SO₄Drying, filtration and concentration under reduced pressure gave the crude material (0.186g), which was purified by preparative HPLC; mobile phase: (A)10mM aqueous ammonium acetate (B) 100% MeCN, column: phenomenex Luna C8(250x21.2) mm, 5 μm, flow rate: 17 ml/min gave the title compound (0.088g, 0.53 mmol). LCMS: method 7, 3.192 min, MS: ES + 166.00; chiral HPLC: method 5, RT 4.81 min, 6.01 min;¹H NMR(400MHz，DMSO-d₆)δppm 7.87(s， 1H)，3.51-3.55(m，1H)，3.38-3.45(m，1H)，3.31-3.36(m，2H)， 3.17-3.21(m，2H)，1.92-2.06(m，3H)，1.80-1.85(m，1H)。

example 63 (R) -6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

Example 62 was subjected to enantiomeric separation using preparative HPLC; mobile phase: (A) 0.1% formic acid in n-hexane (B) 0.1% formic acid IPA solution, column: CHIRALPAK IC SFC (250X21) mm, 5 μm, flow rate: 15 ml/min, two enantiomeric products were obtained, chiral HPLC: method E, 4.53min, 6.00 min. Absolute stereochemistry is named by X-ray crystallography. LCMS: method 7, 3.16 min, MS: ES + 166.0; chiral HPLC: method 5, 4.53 min;¹H NMR(400MHz，DMSO-d₆)δppm 7.87(s，1H)， 3.51-3.55(m，1H)，3.38-3.45(m，1H)，3.31-3.36(m，2H)，3.17 -3.21(m，2H)，1.92-2.06(m，3H)，1.80-1.85(m，1H)。

example 64 (S) -2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ]][1，4]Oxazine-3, 3' -pyrrolidines]-1' -Carbonitrile

Example 49 was subjected to enantiomeric separation using Chiral SFC; mobile phase: (A) liquid carbon dioxide (liq. co)₂) And (B) IPA: MeCN (50: 50), column: CHIRALCEL OJ-H250X 21.0mm, 5 microns, column flow rate of 75.0 ml/min and 100 bar ABPR gave two enantiomeric products, chiral HPLC: column CHIRALART SA 250x4.6mm5 μm, 100% MeOH, 6.82 and 8.37 min. Absolute stereochemistry was named by a similar method as example 63. LCMS: method 2, 3.570 min, MS: ES-305.07; chiral HPLC: column CHIRALART SA 250x4.6mm 5um, 6.82 min, 100% MeOH;¹H NMR(400MHz，DMSO-d₆)δppm 11.27(s，1 H)，8.17(d，J＝2.4Hz，1H)，7.62(d，J＝7.2Hz，2H)，7.48-7.52(m， 3H)，7.41(t，J＝7.2Hz，1H)，3.88(d，J＝11.2Hz，1H)，3.76(d，J＝11.2 Hz，1H)，3.62-3.69(m，2H)，2.38-2.44(m，1H)，2.28-2.32(m，1 H)。

example 75 (S) -7- (3-cyanophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile

Synthesized using 3-cyanophenylboronic acid (CAS number 150255-96-2) using a procedure similar to that described in example 70. LCMS: method 3, 3.598 min, MS: ES + 330.89; chiral HPLC, column CHIRALPAK IC 250 × 4.6mm5 μm mobile phase: IPA: MeCN (50: 50) RT 4.72;¹H NMR(400MHz，DMSO-d6)δppm 10.86(s，1H)， 8.12(s，1H)，8.03(s，1H)，7.87(d，J＝6.8Hz，1H)，7.72-7.82(m， 2H)，7.61-7.65(m，1H)，7.29(s，1H)，3.79-3.82(m，1H)，3.65- 3.76(m，1H)，3.47-3.49(m，1H)，3.48-3.43(m，1H)，2.28-2.35 (m，1H)，1.91-2.03(m，1H)。

scheme 7

Reagents and conditions: a) HATU, DIPEA, THF; b) TFA, DCM; c) TBD, THF; d) pd (OH)₂Polymethylhydroxysilane, (Boc)₂O，EtOH；e)TFA，DCM； f)CNBr，K₂CO₃，THF

Example 76 (8R) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

And a step a. To a solution of Boc-D-alanine (CAS No. 7764-95-6; 1.63g, 8.615 mmol) in THF (32.6ml) at rt was addedHATU (4.09g, 10.775mmol) and DIPEA (3.75ml, 21.55mmol) were added. The reaction mixture was stirred at rt for 1 hour. Methyl 3-amino-1-benzylpyrrolidine-3-carboxylate TFA salt (intermediate E; 2.50g, 7.181mmol) was added to the reaction mixture. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was poured into saturated NaHCO₃Solution (200 ml). The resulting mixture was extracted with EtOAc (2 × 200 ml). The combined organic phases were collected over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (65% EtOAc in hexanes) to give methyl 1-benzyl-3- ((R) -2- ((tert-butoxycarbonyl) amino) propionylamino) pyrrolidine-3-carboxylate (2.30g, 5.675 mmol). LCMS: method 1, 1.67 min, MS: ES + 406.7.

And b, step b. To a solution of methyl 1-benzyl-3- ((R) -2- ((tert-butoxycarbonyl) amino) propionylamino) pyrrolidine-3-carboxylate (2.30g, 5.675mmol) in DCM (23ml) was added TFA (4.6ml) at rt. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was triturated with ether (2 × 10ml) to give the methyl 3- ((R) -2-aminopropionylamino) -1-benzylpyrrolidine-3-carboxylate TFA salt (2.5g, quantitative). This material was used directly in the next step without further purification. LCMS: method 4, 3.542 min, MS: ES + 306.07.

And c, step (c). To a solution of methyl 3- ((R) -2-aminopropionylamino) -1-benzylpyrrolidine-3-carboxylate TFA salt (2.5g, 5.966mmol) in THF (25ml) at rt was added TBD (1.66g, 11.933 mmol). The reaction mixture was stirred at rt for 1.5 h. The resulting reaction mixture was poured into water (150ml) and extracted with EtOAc (3 × 150 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (60% EtOAc in hexane) to afford (8R) -2-benzyl-8-methyl-2, 6, 9-triazaspiro [4.5]]Decane-7, 10-dione (0.90g.3.296 mmol). LCMS: method 3, 3.04 min, MS: ES + 274.5.

And d, step d. In the rt direction (8R) -2-benzyl-8-methyl-2, 6, 9-triazaspiro [4.5]]To a solution of decane-7, 10-dione (0.40g.1.464mmol) in ethanol (8ml) was added 20% Pd (OH)₂(50% moisture) (0.40 g). Poly (methylhydroxysilane) (0.40g) was added dropwise to the reaction mixture at rt, followed by addition (Boc) at rt₂O (0.672ml, 2.928 mmol). The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was combined with another batch of the same scale prepared by the same method, the reaction mixture was filtered through a celite bed and washed with MeOH (3 × 100 ml). The resulting filtrate was concentrated under reduced pressure. The resulting mixture was poured into saturated NaHCO₃(100ml) and extracted with EtOAc (2 × 100 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (4% MeOH in DCM) to give (8R) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5]]Tert-butyl decane-2-carboxylate (0.50g, 1.765 mmol). LCMS: method 1, 1.70 min, MS: ES + 284.2.

And e, step e. To a solution of tert-butyl (8R) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carboxylate (0.20g, 0.706mmol) in DCM (8ml) was added TFA (0.4ml) at rt. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was triturated with ether (2 × 2ml) to give (8R) -8-methyl-2, 6, 9-triazaspiro [4.5] decane-7, 10-dione TFA salt (0.25g, quantitative). This material was used directly in the next step without further purification. LCMS: method 3, 0.803 min, MS: ES + 184.1, 0.88 min.

And f, step d. In the rt direction (8R) -8-methyl-2, 6, 9-triazaspiro [4.5]]To a solution of decane-7, 10-dione TFA salt (0.25g, 0.841mmol) in THF: DMF (9: 1) (10ml) was added K₂CO₃(0.35g, 2.525 mmol). Cyanogen bromide (0.107g, 1.009mmol) was added to the reaction mixture at rt. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was purified by flash chromatography (4.3% MeOH in DCM) to give the title compound as a mixture of diastereomers (0.10g, 0.480 mmol). LCMS: method 3, 1.306 min, MS: ES +209.06, 1.52 min, MS: ES + 209.1;¹H NMR(400MHz，DMSO-d6)δppm8.61(s，2H)，8.40(s，2 H)，4.01-4.04(m，2H)，3.77(d，J＝10Hz，1H)，3.32-3.79(m，7H)， 2.37-2.50(m，1H)，2.26-2.33(m，1H)，1.92-2.06(m，2H)，1.23- 1.28(m，6H)。

example 777, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

Synthesized in step a using Boc-L-glycine using a procedure similar to that described in example 76. LCMS: method 7, 2.89 min, MS: ES + 195.2;¹H NMR(400 MHz，DMSO-d₆)δppm：8.69(s，1H)，8.30(s，1H)，3.83(s，2H)， 0.71(d，J＝10.4Hz，1H)，3.60(q，J＝7.2Hz，1H)，3.51-3.55(m，1H)， 3.47(d，J＝14.8Hz，1H)，2.32-2.41(m，1H)，1.96-2.03(m，1H)。

example 78 (8S) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

Synthesized as a mixture of diastereomers in step a using a procedure similar to that described in example 76, using Boc-L-alanine. LCMS: method 9, 9.39, 9.53 min, MS: ES-207.0;¹H NMR(400MHz，DMSO-d₆)δppm：8.63(d，J＝3.6 Hz，2H)，8.42(s，2H)，4.00-4.39(m，2H)，3.78(d，J＝10Hz，1H)， 3.45-3.66(m，6H)，3.37(d，J＝10Hz，1H)，2.37-2.44(m，1H)，2.26 -2.33(m，1H)，1.93-2.06(m，2H)，1.24-1.28(m，6H)。

example 797, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

Using a similar procedure as described in example 76, 2-, (R) was used in step a(tert-butyloxycarbonyl) amino) -2-phenylacetic acid, synthesized as a mixture of diastereomers. LCMS: method 2, 2.51, 2.63 min, MS: ES-269.4;¹H NMR(400MHz，DMSO-d₆)δppm： 8.86(s，2H)，8.78(s，2H)，7.33-7.42(m，10H)，5.06-5.08(m，2H)， 3.83(d，J＝10.4Hz，1H)，3.74(d，J＝10Hz，1H)，3.50-3.67(m，4H)， 3.47(d，J＝10Hz，1H)，3.22(d，J＝10Hz，1H)，2.44-2.50(m，1H)， 2.33-2.35(m，1H)，2.06-2.11(m，1H)，1.84-1.87(m，1H)。

scheme 8

Reagents and conditions: a) LDA, 2, 3-dibromopropene, THF; b) KRBF₃， PdCl₂(dppf)，Cs₂CO₃Toluene, water; c) k₂OsO₄·2H₂O, sodium periodate, acetone and water; d) NH (NH)₄OAc，NaCNBH₃，EtOH，MgSO₄；e)TFA，DCM；f)CNBr， Na₂CO₃，THF

Example 808-Ethyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

And a step a. To a solution of diisopropylamine (3.72ml, 26.3mmol) in dry THF (30ml) at-78 deg.C was added a solution of 1.6M n-BuLi in hexane (15.4ml, 24.6 mmol). The reaction mixture was stirred at-78 ℃ for 45 minutes. Ethyl 1-Boc-3-pyrrolidinecarboxylate (CAS number 170844-49-2; 2.00g, 8.22mmol) was added to the reaction mixture at-78 ℃ and the reaction mixture was stirred for 1 hour at-78 ℃.2, 3-dibromopropene (CAS number 513-31-5; 1.23 ml, 12.33mmol) was added to the reaction mixture at-78 ℃. The resulting reaction mixture was warmed to 0 ℃. The resulting reaction mixture was combined with another batch of the same scale prepared by the same method and quenched by the addition of saturated ammonium chloride solution (100 ml). By usingThe resulting mixture was extracted with EtOAc (2 × 100 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (5% EtOAc in hexane) to give 1- (tert-butyl) 3-ethyl 3- (2-bromoallyl) pyrrolidine-1, 3-dicarboxylic acid (4.00g, 11.077 mmol). LCMS: method 1, 2.58 min, MS: ES +362.7

And b, step b. To a solution of 3- (2-bromoallyl) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-ethyl ester (0.50g, 1.395mmol) and potassium ethyltrifluoroborate (CAS No. 44248-07-9; 0.23g, 1.661 mmol) in toluene: water (9: 1) (5ml) at rt was added Cs₂CO₃(1.35g, 4.163 mmol). The reaction mixture was degassed with nitrogen at rt for 20min, then PdCl was added at rt₂(dppf) (0.10g, 0.14 mmol). The reaction mixture was heated at 80 ℃ for 15 hours. The resulting reaction mixture was combined with another batch of the same scale prepared by the same method, then cooled to rt, poured into water (100ml) and extracted with EtOAc (2 × 100 ml). The combined organic phases were washed with brine solution (50ml) and Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (8% EtOAc in hexane) to give 1- (tert-butyl) 3-ethyl 3- (2-methylenebutyl) pyrrolidine-1, 3-dicarboxylate (0.32g, 1.028 mmol). LCMS: method 1, 2.886 min, MS: ES + 312.1.

And c, step (c). To a solution of 1- (tert-butyl) 3-ethyl 3- (2-methylenebutyl) pyrrolidine-1, 3-dicarboxylic acid (0.30g, 0.964mmol) in acetone: water (1: 1) (10ml) at rt was added potassium osmate (VI) dihydrate (0.014g, 0.040 mmol). The reaction mixture was cooled to 10 ℃. Sodium metaperiodate (0.83g, 3.840mmol) was added in portions over 15 minutes at 10 ℃. The resulting reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was diluted with water (75ml) and extracted with EtOAc (2 × 75 ml). The combined organic phases were washed with brine solution (50ml) and Na₂SO₄Drying, filtration and concentration under reduced pressure gave 3- (2-oxobutyl) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-ethyl ester (0.30g, 0.957 mmol). LCMS: method 1, 2.313 min, MS: ES +214.18 (M-Boc).

And d, step d. To a solution of 1- (tert-butyl) 3-ethyl 3- (2-oxobutyl) pyrrolidine-1, 3-dicarboxylate (0.30g, 0.957mmol) in ethanol (9ml) at rt was added ammonium acetate (1.11g, 14.388mmol) and NaCNBH₃(0.24g, 3.831 mmol). Addition of MgSO at rt₄(0.81g, 6.715mmol) and the reaction mixture was heated at 80 ℃ for 15 h. The resulting mixture was concentrated under reduced pressure. The resulting residue was dissolved in EtOAc (100mL) and washed with saturated NaHCO₃The solution (50ml), water (50ml), brine solution (50ml) were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (1.5% MeOH in DCM) to afford 8-ethyl-6-oxo-2, 7-diazaspiro [4.4]]Nonane-2-carboxylic acid tert-butyl ester (0.045g, 0.167 mmol). LCMS: method 1, 2.04 min, MS: ES + 213.2 (M-56).

And e, step e. To a solution of tert-butyl 8-ethyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carboxylate (0.04g, 0.159mmol) in DCM (3ml) was added TFA (0.5ml) at rt. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was azeotropically distilled with diethyl ether (5ml) to give 3-ethyl-2, 7-diazaspiro [4.4] nonan-1-one TFA salt (0.03g, 0.106 mmol). MS: ES + 169.2.

And f, step d. To 3-ethyl-2, 7-diazaspiro [4.4] at rt]To a solution of nonan-1-one TFA salt (0.03g, 0.106mmol) in THF (2ml) was added NaHCO₃(0.018g, 0.212 mmol). Cyanogen bromide (0.012g, 0.117mmol) was added to the reaction mixture at rt. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was poured into water (50ml) and extracted with EtOAc (2 × 30 ml). The combined organic phases were collected over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (1% MeOH in DCM) to give the title compound as a mixture of diastereomers (0.008g, 0.041 mmol). LCMS: method 3, 2.78 min, MS: ES + 194.0;¹H NMR(400MHz，CDCl₃)δppm： 5.96(s，2H)，3.80(d，J＝9.6Hz，1H)，3.67-3.73(m，2H)，3.49-3.58 (m，5H)，3.35(d，J＝9.2Hz，1H)，3.27(d，J＝9.6Hz，1H)，2.14-2.43 (m，4H)，1.52-1.92(m，8H)，0.97(t，J＝14.8Hz，6H)。

example 818-benzyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

Synthesized as a mixture of diastereomers in step b using a procedure similar to that described in example 80, using potassium benzyltrifluoroborate. LCMS: method 3, 3.57 min, MS: ES + 356.1;¹H NMR(400MHz，DMSO-d6)δppm：8.12(s，1H)， 7.29-7.31(m，2H)，7.22-7.24(m，3H)，3.78-3.81(m，2H)，3.47-3.52 (m，2H)，3.23(d，J＝9.2Hz，1H)，2.89(dd，J＝4.8Hz，13.2Hz，1H)， 2.58-2.63(m，1H)，1.96-2.05(m，2H)，1.59-1.70(m，2H)。

examples 82 and 838-methyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile

And a step a. A solution of 1- (tert-butyl) 3-ethyl pyrrolidine-1, 3-dicarboxylate (CAS number 170844-49-2; 4.0g, 16.46mmol) in THF (80ml) was cooled to-78 ℃. A1M solution of LiHMDS in THF (21ml, 21.39mmol) was added dropwise to the reaction mixture at-78 ℃. The resulting reaction mixture was stirred at-78 ℃ for 30 minutes. 2- (bromomethyl) prop-1-ene (CAS number 1458-98-6; 3.1g, 23.054mmol) was slowly added to the reaction mixture at-78 ℃. The resulting reaction mixture was warmed to rt. The resulting reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was poured into saturated NH₄Aqueous Cl (20ml) and extracted with EtOAc (3 × 80 ml). The combined organic phases were collected over Na₂SO₄Dried, filtered and concentrated under reduced pressure to give the crude material. The resulting residue was purified by column chromatography (6% EtOAc in hexane) to give 1- (tert-butyl) 3-ethyl 3- (2-methylallyl) pyrrolidine-1, 3-dicarboxylic acid (3.50g, 11.78 mmol). LCMS: in the method 1, the content of the active ingredient,2.63 min, MS: ES +298.4.

And b, step b. A solution of 1- (tert-butyl) 3-ethyl 3- (2-methylallyl) pyrrolidine-1, 3-dicarboxylate (3.50g, 11.78mmol) in MeOH/DCM (1: 1, 40ml) was cooled to-78 ℃. Ozone gas was blown into the reaction mixture at-78 ℃ for 1 hour. The resulting reaction mixture was purged with nitrogen for 10 minutes. Dimethyl sulfide (2.10g, 35.35mmol) was added dropwise to the reaction mixture at-78 ℃. The resulting reaction mixture was warmed to rt and stirred for 1 hour. The reaction mixture was concentrated in vacuo and the resulting residue was purified by column chromatography (18-19% EtOAc in hexanes) to give 1- (tert-butyl) 3-ethyl 3- (2-oxopropyl) pyrrolidine-1, 3-dicarboxylate (1.89g, 6.32 mmol). LCMS: method 1, 2.06 min, MS: ES + 300.3.

And c, step (c). To a solution of 3- (2-oxopropyl) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-ethyl ester (0.50g, 1.67mmol) in THF: acetic acid (9: 1, 10ml) at rt was added CH₃COONH₄(0.64g, 8.36 mmol). The resulting reaction mixture was stirred at rt for 10 min. Sodium triacetoxyborohydride (1.06g, 5.01mmol) was added to the reaction mixture at rt and the resulting reaction mixture was heated at 70 ℃ for 20 hours. The mixture was combined with the other two batches prepared by the same method on the same scale, poured into water (80ml) and washed with solid NaHCO₃And (4) neutralizing. The mixture was extracted with EtOAc (3 × 80ml) and the combined organic phases were collected, over Na₂SO₄Drying, filtration and concentration under reduced pressure gave a residue which was purified by column chromatography (85% EtOAc in hexane) to give 8-methyl-6-oxo-2, 7-diazaspiro [4.4]]Nonane-2-carboxylic acid tert-butyl ester (1.0g, 3.93 mmol). LCMS: method 1, 1.91 min, MS: ES + 255.5.

And d, step d. To a solution of tert-butyl 8-methyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carboxylate (0.4g, 1.57mmol) in DCM (5ml) was added TFA (1.30ml, 15.74 mmol) at 0 ℃. The reaction mixture was stirred at rt for 1 hour. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was triturated with ether (2 × 10ml) to give the TFA salt of 3-methyl-2, 7-diazaspiro [4.4] nonan-1-one (0.40g, quant.). This material was used directly in the next step without further purification. LCMS: method 4, 2.097 min, MS: ES + 155.0.

And e, step e. To 3-methyl-2, 7-diazaspiro [4.4]]To a solution of nonan-1-one TFA salt (0.40g, 1.49 mmol) in THF (5ml) was added K₂CO₃(0.82g, 5.97mmol) to 0 ℃. The reaction mixture was stirred to 0 ℃ for 10 minutes. Cyanogen bromide (0.19g, 1.79mmol) was added to the reaction mixture at 0 ℃ and stirred for another 30 minutes at 0 ℃. The resulting reaction mixture was filtered and concentrated under reduced pressure to give the title compound (0.50g, quantitative) as a crude material. LCMS: method 1, 1.51 min, MS: ES + 180.2. The crude material obtained was subjected to further diastereoisomeric separation by preparative HPLC; mobile phase: (A)20mM aqueous ammonium acetate (B) 100% MeCN: MeOH (50: 50), column: x-bridge C18, 150X19mm, 5 μm, flow rate: 15 ml/min, which gave example 82(0.055g, 0.27mmol) and example 83(0.058g, 0.32 mmol).

Example 828-methyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile: diastereomer 1.

LCMS: method 2, 2.383 min, MS: ES + 179.90; chiral HPLC: method 6, RT 8.25 min, 8.42 min,¹H NMR(400MHz，DMSO-d₆)δppm： 7.97(s，1H)，3.58-3.63(m，1H)，3.35-3.54(m，3H)，3.25-3.28 (m，1H)，2.18-2.23(m，1H)，1.87-1.90(m，2H)，1.54-1.59(m， 1H)，1.11(d，J＝6.0Hz，3H)。

example 838-methyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile: diastereomer 2.

LCMS: method 2, 2.406 min, MS: ES + 179.90; chiral HPLC: method 6, RT 8.16 min, 8.34 min,¹H NMR(400MHz，DMSO-d₆)δppm：7.99(s，1H)，3.52-3.63(m，2H)，3.37-3.42(m，2H)，3.24-3.26 (m，1H)，2.22-2.27(m，1H)，2.07-2.12(m，1H)，1.75-1.78(m， 1H)，1.50-1.55(m，1H)，1.11(d，J＝6.40Hz，3H)。

example 846-oxo-8-phenyl-2, 7-diazaspiro [4.4] nonane-2-carbonitrile: diastereomer 1

Synthesized according to scheme 8. Steps a-e were carried out using a procedure similar to that described in example 80, using potassium phenyltrifluoroborate in step b. Step f and subsequent diastereomer separations were performed using procedures similar to those described for step e of examples 82 and 83. Two racemic diastereomers were obtained, LCMS RT method 2, 3.25 and 3.29 min. One of them shows suitable biological properties and is included as an example. LCMS: method 2, 3.29 min, MS: ES + 242.3;¹H NMR(400MHz，DMSO-d₆)δppm：8.45(s，1H)， 7.27-7.42(m，5H)，4.71(t，J＝7.6Hz，1H)，3.47-3.57(m，2H)，3.35 -3.40(m，2H)，2.55-2.60(m，1H)，2.09-2.17(m，1H)，1.77-1.82(m， 1H)，1.68-1.74(m，1H)。

example 852-oxo-1, 5-dihydro-2H-spiro [ benzo [ e ] [1,4] oxazepine-3, 3 '-pyrrolidine ] -1' -carbonitrile

And a step a. To a solution of 3-hydroxypyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (intermediate C, 0.7g, 2.857mmol) in DMF (10ml) at rt was added K₂CO₃(1.18 g, 8.571mmol) and 2-nitrobenzyl bromide (CAS number 3958-60-9; 0.74g, 3.428 mmol). The reaction mixture was stirred at rt for 16 h, then poured into water (150ml) and extracted with EtOAc (3 × 100 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (neutral alumina, 5% EtOAc in hexanes) to give 1- (tert-butyl) 3-methyl 3- ((2-nitrobenzyl) oxy) pyrrolidine-1, 3-dicarboxylate (0.13g, 0.342 mmol). LCMS: method 1, 2.557 min, MS: ES +325.5 (M-56);¹H NMR(400MHz，DMSO-d₆)δppm：8.04(d， J＝8.0Hz，1H)，7.76(d，J＝4.0Hz，2H)，7.57-7.60(m，1H)，4.78- 4.91(m，2H)，3.71(s，3H)，3.58-3.61(m，2H)，3.41-3.48(m，1H)， 3.33-3.35(m，1H)，2.26-2.28(m，2H)，1.38(d，J＝13.6Hz，9H)。

and b, step b. To a stirred solution of 3- ((2-nitrobenzyl) oxy) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (0.12g, 0.315mmol) in THF: water (1: 1; 6ml) at rt were added iron powder (0.176g, 3.158mmol) and ammonium chloride (0.168g, 3.158 mmol). The reaction mixture was heated at 70 ℃ for 16 hours. The resulting reaction mixture was cooled to rt and filtered through celite hyflow. The celite bed was washed with EtOAc (2 × 10 ml). The combined filtrates were poured into water (50ml) and extracted with EtOAc (3 × 50 ml). The combined organic phases were separated over Na₂SO₄Drying, filtration and concentration under reduced pressure gave 3- ((2-aminobenzyl) oxy) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (0.1g, 0.285 mmol). LCMS: method 1, 2.339 min, MS: ES + 351.38;¹H NMR(400MHz，DMSO-d₆)δppm： 6.99-7.03(m，2H)，6.63(d，J＝8.0Hz，1H)，6.50(t，J＝7.6Hz，1H)， 5.10(s，2H)，4.34-4.38(m，1H)，4.24-4.27(m，1H)，3.75(s，3H)， 3.55-3.69(m，2H)，3.41-3.46(m，1H)，3.29-3.32(m，1H)，2.19 -2.34(m，2H)。1.39(d，J＝4.0Hz，9H)。

and c, step (c). To a stirred solution of 3- ((2-aminobenzyl) oxy) pyrrolidine-1, 3-dicarboxylic acid 1- (tert-butyl) 3-methyl ester (0.08g, 0.228mmol) in THF (5ml) at rt was added TBD (0.064g, 0.457 mmol). The reaction mixture was heated at 70 ℃ for 1 hour. The resulting reaction mixture was cooled to rt, poured into water (50ml) and extracted with EtOAc (2 × 30 ml). The combined organic phases were separated over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The crude material obtained was purified by flash chromatography (3% MeOH in DCM) to give 2-oxo-1, 5-dihydro-2H-spiro [ benzo [ e ]][1，4]Oxazacycloheptatriene-3, 3' -pyrrolidines]-1' -carboxylic acid tert-butyl ester (0.03g, 0.094 mmol). LCMS: method 1, 2.229 min, MS: ES +263.13 (M-56);¹H NMR(400MHz，DMSO-d₆)δppm：10.39(s，1H)，7.21-7.28(m， 2H)，7.12(d，J＝8.0Hz，1H)，6.99-7.03(m，1H)，4.64(s，2H)， 3.61-3.67(m，1H)，3.43-3.57(m，2H)，3.08-3.16(m，1H)，2.28- 2.33(m，1H)，2.14-2.16(m，1H)，1.39(d，J＝4.0Hz，9H)。

and d, step d. To 2-oxo-1, 5-dihydro-2H-spiro [ benzo [ e ] at 0 deg.C][1，4]Oxazacycloheptatriene-3, 3' -pyrrolidines]To a stirred solution of tert-butyl (1' -carboxylate (0.027g, 0.085mmol) in DCM (2ml) was added TFA (0.13 ml). The reaction mixture was stirred at 0 ℃ for 30 minutes. The resulting reaction mixture was concentrated under reduced pressure. The resulting residue was azeotropically distilled with DCM (3 × 5 ml). The resulting material was triturated with hexane (2 × 2ml) and dried under high vacuum to give 1, 5-dihydro-2H-spiro [ benzo [ e ]][1，4]Oxazacycloheptatriene-3, 3' -pyrrolidines]-2-keto TFA salt (0.02g, 0.06 mmol). LCMS: method 1, 1.365 min, MS: ES + 219.28;¹H NMR(400MHz，DMSO-d₆)δppm：10.59(s，1H)，9.20(br，s，2H)， 7.26-7.30(m，2H)，7.15(d，J＝7.6Hz，1H)，7.03-7.05(m，1H)， 4.62-4.72(m，2H)，3.29-3.58(m，4H)，2.27-2.39(m，2H)。

and e, step e. To 1, 5-dihydro-2H-spiro [ benzo [ e ] at 0 DEG C][1，4]Oxazacycloheptatriene-3, 3' -pyrrolidines]To a stirred solution of (0.02g, 0.0602mmol) of the (E) -2-keto TFA salt in THF (5ml) was added K₂CO₃(41.5g, 0.301 mmol). Cyanogen bromide (0.0076g, 0.0723 mmol) was added to the reaction mixture at 0 ℃. The reaction mixture was stirred at 0 ℃ for 10 minutes. The resulting reaction mixture was concentrated in vacuo and the crude material was purified by flash chromatography (2% MeOH in DCM) to give 2-oxo-1, 5-dihydro-2H-spiro [ benzo [ e ] e][1，4]Oxazacycloheptatriene-3, 3' -pyrrolidines]-1' -nitrile (0.009g, 0.037 mmol). LCMS: method 3, 3.414 min, MS: ES + 244.02;¹H NMR(400MHz，DMSO-d₆)δppm： 10.48(s，1H)，7.24-7.29(m，2H)，7.14(d，J＝7.6Hz，1H)，7.02(t， J＝8.0Hz，1H)，4.64-4.72(m，2H)，3.75(d，J＝10.8Hz，1H)，3.57 -3.68(m，2H)，3.43-3.51(m，1H)，2.29-2.37(m，1H)，2.17-2.23 (m，1H)。

example 862-oxo-1, 2,4, 5-tetrahydrospiro [ pyrido [2,3-b ] [1,4] diazepane-3, 3 '-pyrrolidine ] -1' -carbonitrile

And a step a. To a stirred solution of ethyl 2-cyanoacrylate (2.2g, 17.6mmol) in DCM (50ml) at 0 deg.C were added TFA (0.48g, 4.224mmol) and N-benzyl-1-methoxy-N- ((trimethylsilyl) methyl) methylamine (CAS number 93102-05-7; 5.0g, 21.12 mmol). The reaction mixture was stirred at rt for 16 h. The resulting reaction mixture was cooled to 10 ℃ and saturated NaHCO₃The solution (100ml) was quenched. The resulting mixture was extracted with DCM (3 × 50 ml). The combined organic phases were washed with water (50 ml). The organic phase obtained is treated with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (3% EtOAc in hexane) to give ethyl 1-benzyl-3-cyanopyrrolidine-3-carboxylate (2.4g, 9.302 mmol). LCMS: method 1, 1.714 min, MS: ES + 258.8;¹H NMR(400MHz， DMSO-d₆)δppm：7.25-7.36(m，5H)，4.22(q，J＝7.2Hz，2H)，3.67 (dd，J＝13.2Hz，18.0Hz，2H)，3.14(d，J＝9.6Hz，1H)，2.88(d，J＝10.0 Hz，1H)，2.81-2.85(m，1H)，2.53-2.57(m，1H)，2.44-2.47(m， 1H)，2.33-2.40(m，1H)，1.23(t，J＝7.2Hz，3H)。

and b, step b. Raney (Raney) nickel (50% in water) (4.8g, 2.0vol.w/w) was added carefully to a stirred solution of 1-benzyl-3-cyanopyrrolidine-3-carboxylic acid ethyl ester (2.4g, 9.298mmol) in MeOH (25ml) at rt in an autoclave. The reaction mixture was stirred at rt under 250psi hydrogen pressure for 48 hours. The reaction mixture was carefully filtered through a celite bed, washed with MeOH (20ml) and the filtrate was concentrated in vacuo to give a mixture of methyl 3- (aminomethyl) -1-benzylpyrrolidine-3-carboxylate and ethyl 3- (aminomethyl) -1-benzylpyrrolidine-3-carboxylate (2.2g) (44: 56, analysis by LCMS). LCMS: method 4, 3.825 min, 4.133, MS: ES + 249.01, 262.97. The resulting mixture was used directly in the next step without further purification.

And c, step (c). To 3- (aminomethyl) -1-benzylpyrrolidine-3-carboxylic acid methyl ester and 3- (aminomethyl) -1-benzylpyrrolidine-3-carboxylic acid ethyl ester (2.2g, 8.3) at rt96mmol) in toluene (15ml) was added 2-fluoro-3-nitropyridine (1.3g, 9.236mmol) and K₂CO₃(1.74g, 12.595 mmol). The reaction mixture was heated at 120 ℃ for 16 hours. The reaction mixture was cooled to rt, poured into water (150ml) and extracted with EtOAc (3 × 100 ml). The combined organic phases were washed with water (100 ml). Separating the organic phase with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (15% EtOAc in hexane) to give a mixture of methyl 1-benzyl-3- (((3-nitropyridin-2-yl) amino) methyl) pyrrolidine-3-carboxylate and ethyl 1-benzyl-3- (((3-nitropyridin-2-yl) amino) methyl) pyrrolidine-3-carboxylate (1.6 g). LCMS: method 1, 1.782 min, 1.865 min, MS: ES +371.18, 385.18.

And d, step d. Raney nickel (50% in water) (3.2g, 2.0vol.w/w) was carefully added at rt to a stirred solution of a mixture of 1-benzyl-3- (((3-nitropyridin-2-yl) amino) methyl) -pyrrolidine-3-carboxylic acid methyl ester and 1-benzyl-3- (((3-nitropyridin-2-yl) amino) methyl) -pyrrolidine-3-carboxylic acid ethyl ester (1.6g) in MeOH (25 ml). The reaction mixture was purged with hydrogen at rt for 2 hours. The reaction mixture was carefully filtered through a celite bed, washed with MeOH (15ml) and the filtrate was concentrated under vacuum to give a mixture of methyl 3- (((3-aminopyridin-2-yl) amino) methyl) -1-benzylpyrrolidine-3-carboxylate and ethyl 3- (((3-aminopyridin-2-yl) amino) methyl) -1-benzylpyrrolidine-3-carboxylate (1.44 g). LCMS: method 1, 1.434 min, 1.537 min, MS: ES +341.33, 355.34.

And e, step e. A solution of a mixture of methyl 3- (((3-aminopyridin-2-yl) amino) methyl) -1-benzyl-pyrrolidine-3-carboxylate and ethyl 3- (((3-aminopyridin-2-yl) amino) methyl) -1-benzyl-pyrrolidine-3-carboxylate (1.4g) in acetic acid (20ml) was heated at 120 ℃ for 2 h. The reaction mixture was cooled to rt and poured into saturated NaHCO₃Solution (250ml) and extracted with EtOAc (3 × 50 ml). The combined organic phases were separated and washed with DM water (50 ml). Separating the organic phase with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (0.5% MeOH in DCM) to give 1' -benzyl4, 5-dihydrospiro [ pyrido [2,3-b ] yl][1，4]Diazepatriene-3, 3' -pyrrolidines]-2(1H) -one (0.313g, 1.016 mmol). LCMS: method 1, 1.271 min, MS: ES + 309.13;¹H NMR(400MHz， DMSO-d₆)δppm：9.57(s，1H)，7.69(dd，J＝1.2Hz，4.4Hz，1H)， 7.29-7.31(m，4H)，7.23-7.24(m，1H)，7.19(d，J＝7.2Hz，1H)， 6.83-6.88(m，1H)，6.53-6.56(m，1H)，3.51-3.60(m，2H)，3.28 (t，J＝4.8Hz，2H)，2.73(d，J＝9.6Hz，1H)，2.55-2.60(m，2H)， 2.38-2.42(m，1H)，2.03-2.10(m，1H)，1.67-1.72(m，1H)。

and f, step d. To 1' -benzyl-4, 5-dihydrospiro [ pyrido [2,3-b ] at 0 deg.C][1，4]Diazepatriene-3, 3' -pyrrolidines]To a stirred solution of (E) -2(1H) -one (0.31g, 1.006mmol) in THF (5ml) was added K₂CO₃(0.278g, 2.012mmol) and cyanogen bromide (0.107g, 1.006 mmol). The reaction mixture was stirred at rt for 36 hours. The resulting reaction mixture was poured into water (50ml) and extracted with DCM (3 × 25 ml). The combined organic phases were separated and washed with water (50 ml). Separating the organic phase with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by flash chromatography (0.5% MeOH in DCM) to give 2-oxo-1, 2,4, 5-tetrahydrospiro [ pyrido [2,3-b ]][1，4]Diazepatriene-3, 3' -pyrrolidines]-1' -nitrile (0.245g, quantitative). The crude material obtained was purified by preparative HPLC; mobile phase: (a) 100% water and (B) 100% MeCN, column: waters X Bridge C18250X 19mm, 5 microns, flow rate 11.0 ml/min gave 2-oxo-1, 2,4, 5-tetrahydrospiro [ pyrido [2,3-b ]][1，4]Diazepatriene-3, 3' -pyrrolidines]-1' -nitrile (0.077g, 0.317 mmol). LCMS: method 4, 2.939 min, MS: ES + 243.99;¹H NMR(400MHz， DMSO-d₆)δppm：9.85(s，1H)，7.75(dd，J＝1.6Hz，4.8Hz，1H)， 7.28(d，J＝7.6Hz，1H)，6.89(t，4.8Hz，1H)，6.61(dd，J＝4.4Hz， 7.6Hz，1H)，3.69(d，J＝10.0Hz，1H)，3.48(t，J＝7.2Hz，2H)，3.35 -3.36(m，1H)，3.30-3.31(m，1H)，3.20-3.24(m，1H)，2.09-2.16 (m，1H)，1.88-1.95(m，1H)。

example 878-methyl-7, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile

And a step a. To a stirred solution of 2-amino-2-phenylpropionic acid (CAS number 565-07-1; 4.0g, 24.21mmol) in MeCN (50ml) at rt was added tetramethylammonium hydroxide 10% aqueous solution (21.8ml, 24.21 mmol). Boc anhydride (7.92g, 36.0mmol) was added to the reaction mixture at rt. The reaction mixture was stirred at rt for 48 hours. The resulting reaction mixture was poured into water (200ml) and washed with diethyl ether (3 × 25 ml). The aqueous layer was acidified with 5% citric acid solution (-70 ml) and extracted with EtOAc (3 × 30 ml). The combined organic phases were separated over Na₂SO₄Drying, filtration and concentration under reduced pressure gave 2- ((tert-butoxycarbonyl) amino) -2-phenylpropionic acid (4.2g, 15.84 mmol). LCMS: method 1, 2.162 min, MS: ES + 264.28;¹h NMR (400MHz, DMSO-d6) delta ppm: 12.74(s, 1H), 7.43(d, J ═ 8.0Hz, 2H), 7.33(t, J ═ 7.2Hz, 2H), 7.26(t, J ═ 7.2Hz, 1H), 7.14(br s, 1H), 1.72 (s, 3H), 1.36(s, 9H). This material was used directly in the next step without further purification.

And b, step b. To a solution of methyl 2- ((tert-butoxycarbonyl) amino) -2-phenylpropionic acid (1.13g, 4.27mmol) in anhydrous THF (10ml) was added HATU (4.86g, 12.81mmol) and DIPEA (2.2ml, 12.81mmol) at 0 ℃. The reaction mixture was stirred at 0 ℃ for 30 minutes. Methyl 3-amino-1-benzylpyrrolidine-3-carboxylate (intermediate A; 1g, 4.27mmol) was added to the reaction mixture at 0 ℃. The resulting reaction mixture was stirred at rt for 16 h. The resulting reaction mixture was poured into water (20ml) and extracted with DCM (6 × 20 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was purified by column chromatography (50-60% EtOAc in hexanes) to give methyl 1-benzyl-3- (2- ((tert-butoxycarbonyl) amino) -2-phenylpropionylamino) pyrrolidine-3-carboxylate (2.59g, quantitative). LCMS: method 1, 2.015 min, MS: ES + 482.28.

And c, step (c). To a stirred solution of methyl 1-benzyl-3- (2- ((tert-butoxycarbonyl) amino) -2-phenylpropionylamino) pyrrolidine-3-carboxylate (1.2g, 2.49mmol) in DCM (5ml) was added TFA (12ml) at 0 ℃. The reaction mixture was stirred at rt for 3 hours. The resulting reaction mixture was concentrated under vacuum. The resulting residue was triturated with n-pentane (3 × 10ml) and dried under high vacuum. The crude material obtained was quickly poured into saturated NaHCO₃The solution was stirred well until pH 7-8. The resulting mixture was extracted with a mixture of MeOH: DCM (1: 9, 5X 15 ml). The combined organic phases were washed with Na₂SO₄Drying, filtration and concentration under reduced pressure gave methyl 3- (2-amino-2-phenylpropionylamino) -1-benzylpyrrolidine-3-carboxylate (0.776g, 2.036 mmol). LCMS: method 3, 4.167 min, MS: ES + 382.05. This material was used directly in the next step without further purification.

And d, step d. To a solution of methyl 3- (2-amino-2-phenylpropionylamino) -1-benzylpyrrolidine-3-carboxylate (0.776g, 2.036mmol) in dry THF (10ml) at rt was added TBD (0.708 g, 5.09 mmol). The reaction mixture was stirred at rt for 12 hours. The resulting reaction mixture was poured into water (20ml) and extracted with EtOAc (3 × 40 ml). The combined organic phases were washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The resulting residue was triturated with ether (2 × 15ml) and dried under vacuum to give 2-benzyl-8-methyl-8-phenyl-2, 6, 9-triazaspiro [4.5]]Decane-7, 10-dione (0.380g, 1.08 mmol). LCMS: method 1, 1.586 min, MS: ES + 350.48. This material was used directly in the next step without further purification.

And e, step e. In the rt direction 2-benzyl-8-methyl-8-phenyl-2, 6, 9-triazaspiro [4.5]]To a stirred solution of decane-7, 10-dione (0.380g, 1.08mmol) in MeOH (15ml) was added acetic acid (2 ml). Pd/C (50% moisture) (65mg) was added to the reaction mixture at rt. Purging the reaction mixture at rt H₂For 1.5 hours. The reaction mixture was carefully filtered through celite hyflow. The celite bed was washed with MeOH (2 × 5 ml). The combined filtrates were concentrated in vacuo. The resulting residue was triturated with ether (2 × 10ml) and dried under high vacuum to give 8-methyl-8-phenyl-2, 69-triazaspiro [4.5]Decane-7, 10-dione (0.384g, 1.203 mmol). LCMS: method 3, 2.338, 2.551 min, MS: ES + 259.98. This material was used directly in the next step without further purification.

And f, step d. In the rt orientation 8-methyl-8-phenyl-2, 6, 9-triazaspiro [4.5]]To a stirred solution of decane-7, 10-dione (0.384g, 1.482mmol) in THF: DMF (2: 1, 8ml) was added K₂CO₃(0.519g, 3.764 mmol). The reaction mixture was stirred at rt for 15 minutes. Cyanogen bromide (0.159g, 1.504mmol) was added to the reaction mixture. The reaction mixture was stirred at rt for 45 minutes. The resulting reaction mixture was poured into ice-cold water (30 ml). The resulting precipitate was filtered under vacuum, washed with water (20ml) and air dried. The resulting solid material was triturated with ether (3 × 10ml) and dried under high vacuum to give 8-methyl-7, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5]]Decane-2-carbonitrile (0.148g, 0.520 mmol). LCMS: method 2, 2.820, 2.907 min, MS: ES +285.24, 285.29;¹H NMR(400MHz， DMSO-d6)δppm：9.18(s，1H)，8.77(s，1H)，7.40-7.43(m，5H)， 3.61-3.63(m，1H)，3.33-3.51(m，2H)，3.17-3.18(m，1H)，1.97 -2.01(m，1H)，1.81-1.85(m，1H)，1.60(s，3H)。

biological Activity of the Compounds of the invention

Abbreviations:

TAMRA carboxy tetramethyl rhodamine

PCR polymerase chain reaction

PBS phosphate buffered saline

EDTA ethylene diamine tetraacetic acid

Tris 2-amino-2- (hydroxymethyl) -1, 3-propanediol

NP-40 Nonidet P-40, octylphenoxy polyethoxyethanol

BSA bovine serum Albumin

DMSO dimethyl sulfoxide

In vitro inhibition assay for Cezanne1

Expression and purification of Cezanne1

The Cezanne1 construct was PCR amplified and cloned into pFLAG-CMV-6c vector (Sigma-Aldrich) with an N-terminal FLAG marker. HEK293T cells were transfected with FLAG-Cezanne 1 using TransIT-LT1 transfection reagent (Mirus) according to the manufacturer's instructions. Cells were harvested 48 hours after transfection. The cells were washed once with PBS and scraped in lysis buffer (50mM Tris, pH7.5, 150mM NaCl, 3mM EDTA, 0.5% NP40, 10% glycerol, 5mM beta-mercaptoethanol, protease inhibitors (complete mini, Roche) and phosphatase inhibitors (PhosSTOP mini, Roche.) the lysates were incubated on ice for 30 minutes and centrifuged at 4000rpm for 10 minutes at 4 ℃, the soluble supernatant was added to FLAG affinity resin (EZview Red ANTI-FLAG M2 affinity gel, Sigma-Aldrich) equilibrated in low salt buffer (20mM Tris, pH7.5, 150mM NaCl, 0.5mM EDTA, 5mM beta-mercaptoethanol) and incubated at 4 ℃ for 3 hours, the resin was spun at 2000rpm for 2 minutes, and the Tris supernatant was removed the resin was washed 2 times with low salt buffer and washed with high salt buffer (20mM, pH7.5, 500mM NaCl, 0.5mM EDTA, 5mM β -mercaptoethanol, protease inhibitors (complete mini, Roche) and phosphatase inhibitors (PhosSTOP mini, Roche) were washed once, and to elute the bound Cezanne1, an elution buffer (10mM Tris, pH7.5, 150mM NaCl, 0.5mM EDTA, 10% glycerol, 0.5% NP40, 5mM β -mercaptoethanol, 0.15mg/ml 3X FLAG peptide (Sigma-Aldrich)) was added to the resin and incubated at 4 ℃ for 2.5 hours while spinning. The resin was centrifuged at 4000rpm for 30 seconds and the supernatant containing the purified FLAG-Cezanne 1 was removed and stored at-80 ℃.

The molecular weight of the purified FLAG-protein was characterized relative to BSA using SDS-PAGE as shown in figure 1 (left hand scale in kilodaltons).

Cezanne1 Biochemical kinetics assay

The reaction was performed in duplicate in a black 384-well plate (small volume, Greiner 784076) at a final reaction volume of 21 μ l. Cezanne1 was diluted in reaction buffer (40mM Tris, pH7.5, 0.005% Tween 20, 0.5mg/ml BSA, 5 mM-. beta. -mercaptoethanol) to equivalent concentrations of 0, 0.001, 0.050, 0.01, and 0.05. mu.l/well. The buffer is optimized for optimal temperature, pH, reducing agents, salts, incubation time and detergents. The reaction was initiated by adding 50nM of TAMRA-tagged peptide linked to ubiquitin via an isopeptide bond as a fluorescence polarizing substrate. The reaction was incubated at room temperature and read 1 time every 2min for 120 min. The reading was performed with the Pherastar Plus (BMG Labtech). λ excitation 540 nm; lambda emits at 590 nm. Figure 2 shows a graph of the proteolytic activity of Cezanne1 measured using fluorescence polarization assay. Different volumes of purified Cezanne1 as indicated were incubated with TAMRA-labeled peptides linked to ubiquitin via isopeptide bonds.

Cezanne1 Biochemical IC50 assay

Dilution plates were prepared in 96-well polypropylene V-bottom plates (Greiner #651201) in 50% DMSO at 21-fold final concentration (2100 μ M, final concentration 100 μ M). Typical 8-point dilution series are 100, 30, 10, 3, 1, 0.3, 0.1, 0.03 μ M final concentration. Reactions were performed in duplicate in a final reaction volume of 21 μ l in black 384-well plates (small volume, Greiner 784076). Add 1. mu.l of 50% DMSO or diluted compound to the plate. Cezanne1 was diluted in reaction buffer (40mM Tris, pH7.5, 0.005% Tween 20, 0.5mg/ml BSA, 5 mM. beta. -mercaptoethanol) to an equivalent concentration of 0.005. mu.l/well, and 10. mu.l of the diluted Cezanne1 was added to the compound. The enzyme and compound were incubated at room temperature for 30 min. The reaction was initiated by adding 50nM of TAMRA-tagged peptide linked to ubiquitin via an isopeptide bond as a fluorescence polarizing substrate. The reaction was read immediately after addition of substrate and 2 hours incubation at room temperature. The reading was performed with the Pherastar Plus (BMG Labtech). λ excitation 540 nm; lambda emits at 590 nm.

Activity of exemplary compounds in the Cezanne1 biochemical IC50 assay.

The range is as follows:

A＜0.1μM；

0.1＜B＜1μM；

1＜C＜10μM；

D＞10μM

Claims (16)

1. a compound of formula I:

a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, wherein:

R^1a、R^1b、R^1cand R^1dEach independently represents hydrogen or C₁-C₆An alkyl group;

R^1eand R^1fEach independently represents hydrogen or C₁-C₆An alkyl group;

ring A is selected from the group consisting of piperidin-2-one, piperazin-2-one, pyrrolidin-2-one, indolin-2-one, 3, 4-dihydroquinolin-2 (1H) -one, 1H-pyrido [2,3-b ] [1,4] oxazin-2 (3H) -one, 3, 4-dihydropyrido [2,3-b ] pyrazin-2 (1H) -one, 1, 5-dihydrobenzo [ e ] [1,4] oxazepine-2 (3H) -one, 3, 4-dihydro-1, 5-naphthyridin-2 (1H) -one, 3, 4-dihydro-1, 6-naphthyridin-2 (1H) -one, 3, 4-dihydro-1, 7-naphthyridin-2 (1H) -one, 3, 4-dihydro-1, 8-naphthyridin-2 (1H) -one, 3, 4-dihydropyrazino [2,3-b ] pyrazin-2 (1H) -one, and 1,2,3, 5-tetrahydro-4H-pyrido [2,3-b ] [1,4] diazepin-2-one;

wherein ring A is unsubstituted or substituted by one or two-Q¹-(R²)_nSubstituted, each of which is-Q¹-(R²)_nThe same or different, and wherein:

n is 0 or 1;

when n is 0, Q¹Selected from halogen, cyano, oxo, -CONR³R⁴、-C₁-C₆Alkyl and-C₁-C₆An alkoxy group; wherein said alkyl is unsubstituted or substituted with one to four halogens;

when n is 1, Q¹Is selected fromCovalent bond, -CO-, -CONR³-C₀-C₃Alkylene and C₁-C₆An alkylene group;

R²selected from the group consisting of piperidinyl, pyrrolyl, phenyl, pyrazolyl, isoxazolyl, indazolyl, pyridinyl, dihydropyridinyl, benzothiazolyl, and pyrimidinyl;

wherein R is²Unsubstituted or substituted by one to four substituents selected from halogen, cyano, oxo, -CONR⁶R⁷、-NR⁶COR⁷、-C₁-C₆Alkyl, -C₁-C₆Alkoxy, -Q^2a-R⁸、-Q^2b-CONR⁶-Q^2c-R⁸and-Q²-NR⁶SO₂-Q^2c-R⁸(ii) a Wherein said alkyl is unsubstituted or substituted with one hydroxy or one to four halogens;

Q^2arepresents a covalent bond, an oxygen atom, -SO₂-, -CO-or C₁-C₆An alkylene group;

Q^2band Q^2cEach represents a covalent bond;

R³and R⁴Each independently represents hydrogen or C₁-C₆An alkyl group;

R⁶and R⁷Each independently represents hydrogen or C₁-C₆An alkyl group; and

R⁸selected from phenyl, piperazinyl, cyclopropyl, morpholinyl and piperidinyl; wherein R is⁸Optionally substituted by fluorine, chlorine, oxo, cyano, C₁-C₃Alkyl or C₁-C₃Alkoxy substitution.

2. A compound according to claim 1, wherein ring a is selected from the group consisting of 3, 4-dihydroquinolin-2 (1H) -one, 1H-pyrido [2,3-b ] [1,4] oxazin-2 (3H) -one, 3, 4-dihydropyrido [2,3-b ] pyrazin-2 (1H) -one, 1, 5-dihydrobenzo [ e ] [1,4] oxazepine-2 (3H) -one, and 1,2,3, 5-tetrahydro-4H-pyrido [2,3-b ] [1,4] diazepin-2-one.

3. The compound according to claim 1, wherein ring a is selected from the group consisting of piperazin-2-one and pyrrolidin-2-one.

4. A compound according to any one of claims 1 to 3, wherein n is 1; and Q¹Selected from the group consisting of covalent bond and C₁-C₃An alkylene group.

5. A compound according to any one of claims 1 to 3, wherein R²Unsubstituted or substituted by one or two substituents selected from halogen, cyano, oxo, C₁-C₃Alkyl radical, C₁-C₃Alkoxy, -CONR⁶R⁷、-NR⁶COR⁷、-Q^2a-R⁸and-Q^2b-NR⁶SO₂-Q^2c-R⁸(ii) a Wherein alkyl is unsubstituted or substituted by fluoro;

Q^2ais a covalent bond, an oxygen atom, -CO-, -SO₂-or-C₁-C₃An alkylene group;

Q^2bis a covalent bond;

Q^2cis a covalent bond;

R⁶and R⁷Each independently selected from hydrogen and C₁-C₃An alkyl group; and

R⁸selected from phenyl, piperazinyl, cyclopropyl, morpholinyl and piperidinyl.

6. A compound according to any one of claims 1 to 3, wherein n is 0; q¹Selected from oxo, methyl, ethyl, CF₃Methoxy, halogen and-C (O) NR³R⁴(ii) a And R is³And R⁴Each independently selected from hydrogen and methyl.

7. A compound according to any one of claims 1 to 3, wherein R^1a、R^1b、R^1c、R^1d、R^1fAnd R^1eEach is hydrogen.

8. A compound according to claim 1 selected from the group consisting of:

2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7 '-chloro-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7 '-methoxy-2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (5-isopropyl-2-methoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7' - ([1,1' -biphenyl ] -4-yl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (4- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (2-fluoro-5-methylphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (3-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (1-methyl-1H-pyrazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

2 '-oxo-7' - (4-phenoxyphenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (1-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (4-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (2-chloro-5- (trifluoromethoxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

5- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) -N-methylpyridine carboxamide;

7'- (2- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) -N-methylbenzamide;

7'- (3- ((2-chlorobenzyl) oxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (4- (4-methylpiperazin-1-yl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (6-methoxypyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (5-fluoro-2-isopropoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (3-methyl-1H-indazol-6-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (4- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (1-methyl-1H-indazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (5-methyl-1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

n- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) cyclopropanesulfonamide;

7'- (3-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

2 '-oxo-7' - (pyrimidin-5-yl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

n- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide;

3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) -N, N-dimethylbenzamide;

n- (4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide;

7'- (4- (morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (3, 5-dimethyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (2-methylpyridin-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

2 '-oxo-7' - (3- (piperidin-1-yl) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

n- (2- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) phenyl) acetamide;

7'- (4- (morpholine-4-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (3- (morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (1-methyl-6-oxo-1, 6-dihydropyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (2-methylbenzo [ d ] thiazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

2 '-oxo-6' -phenyl-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (4-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (3-cyanophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (4-fluorophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (3-fluorophenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

1-cyano-N, N-dimethyl-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -carboxamide;

1-cyano-N-methyl-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinoline ] -7' -carboxamide;

2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (4-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (3-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (3-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

2-oxo-6-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

2-oxo-6- (trifluoromethyl) -1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

2-oxo-7-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (4-cyanophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (4-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (3-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

3-oxo-3, 4-dihydro-1H-spiro [ pyrido [2,3-b ] pyrazine-2, 3 '-pyrrolidine ] -1' -carbonitrile;

(R) -6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile;

(S) -2-oxo-7-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(R) -2 '-oxo-6' -phenyl-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

(S) -2-oxo-7-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (3-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (4-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (3-cyanophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (4-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (3-fluorophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (3-cyanophenyl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(8R) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile;

7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile;

(8S) -8-methyl-7, 10-dioxo-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile;

7, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile;

8-benzyl-6-oxo-2, 7-diazaspiro [4.4] nonane-2-carbonitrile;

2-oxo-1, 5-dihydro-2H-spiro [ benzo [ e ] [1,4] oxazepine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

2-oxo-1, 2,4, 5-tetrahydrospiro [ pyrido [2,3-b ] [1,4] diazepane-3, 3 '-pyrrolidine ] -1' -carbonitrile;

8-methyl-7, 10-dioxo-8-phenyl-2, 6, 9-triazaspiro [4.5] decane-2-carbonitrile;

2-oxo-6-phenyl-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (5-methyl-1H-indazol-4-yl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (1, 4-dimethyl-1H-pyrazol-5-yl) -2-oxo-1, 4-dihydro-2H-spiro [ pyrido [2,3-b ] pyrazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(R) -7'- (5-methyl-1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

(R) -7'- (4- (4-methylpiperazin-1-yl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

(R) -7'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

(S) -7'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

(R) -6'- (1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

1 '-cyano-N- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3' -pyrrolidine ] -6-carboxamide;

2-oxo-6- (piperidine-1-carbonyl) -1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

6- (1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -6- (1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -1 '-cyano-N- (4-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3' -pyrrolidine ] -6-carboxamide;

1 '-cyano-2-oxo-N-phenyl-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3' -pyrrolidine ] -6-carboxamide;

1 '-cyano-N- (2-fluorophenyl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3' -pyrrolidine ] -6-carboxamide;

7- (1-methyl-1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(R) -7- (1-methyl-1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (1-methyl-1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (1- (2-hydroxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(R) -7- (1- (2-hydroxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (1- (2-hydroxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (1- (2-methoxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(R) -7- (1- (2-methoxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (1- (2-methoxyethyl) -1H-indazol-4-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

7- (6-methoxy-2-methylpyridin-3-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(R) -7- (6-methoxy-2-methylpyridin-3-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

(S) -7- (6-methoxy-2-methylpyridin-3-yl) -2-oxo-1, 2-dihydrospiro [ pyrido [2,3-b ] [1,4] oxazine-3, 3 '-pyrrolidine ] -1' -carbonitrile;

2 '-oxo-7' - (3- (trifluoromethoxy) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -7' -yl) -N, N-dimethylbenzamide;

7'- (3- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

7'- (1-methyl-1H-pyrrol-2-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6' - ([1,1' -biphenyl ] -4-yl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (4- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (1-methyl-1H-pyrazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

2 '-oxo-6' - (3- (trifluoromethoxy) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

2 '-oxo-6' - (4-phenoxyphenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (1-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

5- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) -N-methylpyridine carboxamide;

6'- (2- (benzyloxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) -N-methylbenzamide;

6'- (5-isopropyl-2-methoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (3- ((2-chlorobenzyl) oxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (6-methoxypyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (5-fluoro-2-isopropoxyphenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (3-methyl-1H-indazol-6-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (4- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (1-methyl-1H-indazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (5-methyl-1H-indazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

n- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) phenyl) cyclopropanesulfonamide;

4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) -N, N-dimethylbenzamide;

2 '-oxo-6' - (pyrimidin-5-yl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

n- (3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) phenyl) acetamide;

n- (4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) phenyl) acetamide;

6'- (3- (4-methylpiperazine-1-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (1-methyl-1H-pyrrol-2-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (4- (morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (3, 5-dimethyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

2 '-oxo-6' - (3- (piperidin-1-yl) phenyl) -1',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

n- (2- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) phenyl) acetamide;

6'- (4- (morpholine-4-carbonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (3- (morpholinosulfonyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (1-methyl-6-oxo-1, 6-dihydropyridin-3-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (2-methylbenzo [ d ] thiazol-5-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6' - (3, 5-dimethyliso)Azol-4-yl) -2' -oxo-1 ',4' -dihydro-2 ' H-spiro [ pyrrolidine-3, 3' -quinoline]-1-nitrile;

6'- (2-chloro-5- (trifluoromethoxy) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (4- (4-methylpiperazin-1-yl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

n-benzyl-4- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) benzamide;

6'- (3-methyl-1H-pyrazol-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

6'- (4- (morpholinomethyl) phenyl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

3- (1-cyano-2 '-oxo-1', 4 '-dihydro-2' H-spiro [ pyrrolidine-3, 3 '-quinolin ] -6' -yl) -N, N-dimethylbenzamide; and

6'- (2-methylpyridin-4-yl) -2' -oxo-1 ',4' -dihydro-2 'H-spiro [ pyrrolidine-3, 3' -quinoline ] -1-carbonitrile;

a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer.

9. Use of a compound of formula (I), a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, according to any one of claims 1 to 8, for the manufacture of a medicament for the treatment of cancer, inflammation, or a condition involving mitochondrial dysfunction.

10. The use according to claim 9, wherein the cancer is selected from the group consisting of breast cancer, ovarian cancer, prostate cancer, lung cancer, kidney cancer, stomach cancer, colon cancer, testicular cancer, head and neck cancer, pancreatic cancer, brain cancer, melanoma, bone cancer, leukemia, and multiple myeloma.

11. The use according to claim 9, wherein said cancer is selected from the group consisting of cancer of tissue and organs, cancer of blood cells and lymphoma.

12. The use according to claim 9, wherein the condition involving mitochondrial dysfunction is selected from CNS disorders; neurodegenerative diseases; amyotrophic lateral sclerosis; ischemia; stroke; multiple sclerosis; mitochondrial encephalopathy, lactic acidosis and stroke-like seizure syndrome; leber's hereditary optic neuropathy; cancer; ataxia; retinitis pigmentosa-maternal hereditary acute necrotizing encephalopathy; danon's disease; diabetes mellitus; diabetic nephropathy; metabolic disorders; heart failure; ischemic heart disease leading to myocardial infarction; psychosis; multiple sulfatase deficiency; mucolipidosis II; mucolipidosis III; mucolipidosis IV; GMl-gangliosidosis; neuronal ceroid-lipofuscinosis; alpes disease; barth syndrome; beta-oxidation defects; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency syndrome; coenzyme Q10 deficiency; deficiency of Complex I; complex II deficiency; complex III deficiency; a deficiency of complex IV; complex V deficiency; COX deficiency; chronic progressive lateral ophthalmoplegic syndrome; CPTI deficiency; CPTII deficiency; glutaric aciduria type II; Bass-Seldii syndrome; lactic acidosis; a long-chain acyl-CoA dehydrogenase deficiency; a li's disease or syndrome; fatal infantile cardiomyopathy; left disease; glutaric aciduria type II; medium chain acyl-CoA dehydrogenase deficiency; myoclonic epilepsy and fluffy-hairlike red cellulose syndrome; mitochondrial cell disease; mitochondrial degenerative ataxia syndrome; mitochondrial DNA depletion syndrome; muscular neurogastrointestinal disorders and encephalopathy; pearson syndrome; pyruvate dehydrogenase deficiency; pyruvate carboxylase deficiency; POLG mutation; medium/short chain 3-hydroxyacyl-CoA dehydrogenase deficiency; and very long chain acyl-CoA dehydrogenase deficiency; and age-dependent decline in cognitive function and muscle strength.

13. The use according to claim 9, wherein the condition involving mitochondrial dysfunction is a neuropathy.

14. Use according to claim 12, wherein said neurodegenerative disease is selected from parkinson's disease, alzheimer's disease, huntington's disease, dementia with lewy bodies and frontotemporal dementia.

15. Use according to claim 12, wherein the psychosis is schizophrenia.

16. A pharmaceutical composition comprising a compound of formula (I), a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, according to any one of claims 1 to 8, and one or more pharmaceutically acceptable excipients.