CN114230569A

CN114230569A - Thiohydrazide compound, preparation method and application thereof

Info

Publication number: CN114230569A
Application number: CN202111042494.6A
Authority: CN
Inventors: 杨千姣; 周游; 潘德思; 山松; 王晓亮; 宋永连; 李志斌; 鲁先平
Original assignee: Shenzhen Chipscreen Biosciences Co Ltd
Current assignee: Shenzhen Chipscreen Biosciences Co Ltd
Priority date: 2020-09-09
Filing date: 2021-09-07
Publication date: 2022-03-25

Abstract

The invention relates to a thio compoundHydrazide compounds, a preparation method and application thereof. The structure of the compound is shown as a formula (I), and the definitions of all variables in the formula are described in the specification. The compounds are a novel class of p53 activity modulators. The invention also provides a pharmaceutical composition containing the compound as an active ingredient, which can be used for treating and/or preventing diseases related to p53 protein mutation and has important application value.

Description

Thiohydrazide compound, preparation method and application thereof

Technical Field

The invention belongs to the technical field of medicines, and relates to a novel thiohydrazide compound of a p53 activity regulator and an isomer thereof, a preparation method of the compound, a pharmaceutical composition containing the compound as an active ingredient, and pharmaceutical application thereof. The compound of the invention can be used as an activity regulator targeting p53, and is used for treating and/or preventing tumors or cancers related to p53 protein mutation.

Background

The p53 gene was first discovered since 1979 and has been the focus of research. In 1993, the journal of Science rated it as an "annual molecule". The p53 gene coded protein, namely p53 protein, the monomer of which is composed of 393 amino acid residues and is in a cross-shaped structure, and the protein comprises an N-terminal unfolded trans-activation domain (TAD) and a proline enrichment region (PRD), and the function of the protein is mainly to play a role in transcriptional activation; the C-terminus is a flexible unfolded region (CT) whose function is mainly to participate in the formation of tetramers of proteins; the central core region is the DNA Binding Domain (DBD) which binds to specific DNA sequences and thus exerts normal physiological functions (Demir O, Ieong PU, Amaro RE. full-length p53 tetramer bound to DNA and its quantitative genes. oncogene.2017,36(10): 1451-.

The p53 protein is one of The most important tumor suppressors to date and one of The most effective "natural killers" against cancer in human cells, has transcription factor activity, and can form complexes with other proteins to exert different biological functions, including playing a key role in The cycle regulation of cell growth and development, DNA repair, senescence or apoptosis (Vousden KH, scenes C. Blindedby The Light: The growing compatibility of p53.cell.2009,137(3): 413-431.).

However, p53 protein is also one of the most easily mutated proteins, and after the gene is mutated, its structural spatial conformation is changed, and its original normal biological function is lost, so that the p53 gene is converted into cancer gene from cancer suppressor gene. Research shows that p53 gene mutation is found in more than half of human tumor tissues, which is the most common genetic mutation in tumors, and the mutation types comprise nonsense mutation, displacement mutation, missense mutation and the like, but the missense mutation is still the main mutation. Hot-spot mutations of p53 in human tumors are mainly concentrated in highly conserved DNA Binding Domains (DBDs), including sites R175, G245, R248, R249, R273, R282, etc., which upon mutation result in changes in spatial conformation of p53 protein, thereby altering its stability in binding to DNA, thereby altering the normal biological functions of p53 protein, such as promoting the process of tumor development and development (Brosh R, rotator V.When mutations gain new powers: new from the mutant p53 field. Nat Rev cancer.2009,9(10): 701-). In general, p53 protein mutations are mostly point mutations, and small molecule compounds can restore the weak conformational change, which provides important scientific basis for designing small molecule compounds to restore the p53 conformation. Therefore, drug development of small molecule activity modulators based on mutant p53 proteins would bring significant benefits to tumor patients.

Up to now, the publicly reported small molecule active compounds against mutant p53 protein are mainly COTI-2, ZMC1, PRIMA-1MET (APR-246), CP-31398, WR1065, STIM-1, and MIRA-1, and the clinical research is slow in progress, and there is no approved drug on the market at present all over the world. Therefore, we expect to develop new modulators of activity targeting p53 to meet unmet clinical needs.

Disclosure of Invention

In one aspect, the invention relates to thiohydrazide compounds, including isomers thereof, capable of modulating the activity of p53 protein.

Another aspect of the invention relates to methods of making the compounds described herein.

Yet another aspect of the present invention relates to pharmaceutical compositions comprising the compounds of the present invention as active ingredients, and the clinical use of the compounds or pharmaceutical compositions of the present invention for the treatment and/or prevention of diseases associated with a targeted p53 protein, in particular for the treatment of tumors or cancers.

The invention relates to a compound shown in a formula (I), comprising pharmaceutically acceptable salt or isomer thereof,

wherein the content of the first and second substances,

R¹selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl (e.g., furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, or triazolyl, preferably pyridyl), wherein said heteroaryl may be independently selected from deuterium, halo (e.g., fluoro, chloro, bromo, or iodo), C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino radicalDouble C₁-C₄Alkylamino, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring;

R²selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl (e.g., furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, or triazolyl, preferably pyridyl), wherein said heteroaryl may be independently selected from deuterium, halo (e.g., fluoro, chloro, bromo, or iodo), C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring;

alternatively, the first and second electrodes may be,

R¹and R²Together with the carbon atom to which they are commonly attached form a heteroaryl (e.g., dihydroquinolinyl) or 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from the group consisting of halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to aryl or heteroaryl, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle;

R³selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, aryl, arylamino, heteroaryl, heteroarylamino, heteroarylaminoaryl(s)Heteroaryl such as furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl; such as phenyl), a 3-15 membered mono-or polycyclic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen or sulfur (e.g. azaspiro [3.3]]Heptane-6-yl), 3-15 membered mono-or polycyclic heterocyclic amino, wherein said arylamino, heteroarylamino, 3-15 membered mono-or polycyclic heterocyclic amino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

R⁴selected from hydrogen, deuterium, C₁-C₄Alkylamino, halogeno C₁-C₄Alkylamino, arylamino, heteroarylamino, heteroarylaminoaryl (said heteroaryl being, for example, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl; said aryl being, for example, phenyl), 3-15 membered mono-or polycyclic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen or sulfur (for example, azaspiro [3.3]]Heptane-6-yl), 3-15 membered mono-or polycyclic heterocyclic amino, wherein said C₁-C₄Alkylamino, halogeno C₁-C₄Alkylamino, arylamino, heteroarylamino, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

R³and R⁴Together with the nitrogen atom to which they are jointly attached form an optionalA 3-15 membered mono-or polycyclic heterocycle containing one additional heteroatom selected from nitrogen, oxygen or sulfur, wherein said 3-15 membered mono-or polycyclic heterocycle may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

R³and R⁴Exemplary 3-15 membered mono-or polycyclic heterocycles formed with the nitrogen atom to which they are commonly attached include, but are not limited to:

R^aselected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, aryl, heteroaryl (e.g., furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl; benzofuryl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzotriazolyl, benzoxazolyl, furopyridyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuryl, isobenzothienyl, isoindolyl, isoquinolyl, dihydroquinolinyl, naphthyridinyl, oxazolopyridyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quinazolinyl, pyrazinyl, pyrazolyl, pyridazinyl, benzothiazolyl, benzimidazolyl, benzoxazolyl, benzothienyl, isoindolyl, isoquinolinyl, dihydroquinolinyl, naphthyridinyl, oxazolopyridyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolyl, and the like, Thiadiazolopyrimidinyl and thienopyridinyl; preferably pyridyl, quinolyl), heteroarylamino (said heteroaryl being for example furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl), wherein said aryl, heteroaryl may be one or more independently selected from the group consisting ofHydrogen, deuterium, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkoxy, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino radical, mono C₁-C₄Alkylamino radical C₁-C₄Alkyl, di-C₁-C₄Alkylamino radical C₁-C₄Alkyl, and the heteroaryl is a 5-12 membered monocyclic or polycyclic ring;

in a preferred aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, including isomers thereof, wherein,

R¹selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl, wherein said heteroaryl may be independently selected from deuterium, halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring;

R²selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl, wherein said heteroaryl may be independently selected from deuterium, halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring;

alternatively, the first and second electrodes may be,

R³selected from hydrogen, deuterium;

R⁴selected from arylamino, heteroarylamino, heteroarylaminoaryl (said heteroaryl being, for example, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl; said aryl being, for example, phenyl), 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino, wherein said arylamino, heteroarylamino, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

R³and R⁴Together with the nitrogen atom to which they are both attached form a 3-15 membered mono-or polycyclic heterocycle, wherein the 3-15 membered mono-or polycyclic heterocycle may be substituted with one or more R^aSubstituted, said 3-15 membered polycyclic heterocycle includes but is not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused toAryl or heteroaryl;

R^aselected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, aryl, heteroaryl, heteroarylamino (said heteroaryl being for example furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl), wherein said aryl, heteroaryl may be substituted with one or more groups independently selected from hydrogen, deuterium, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkoxy, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino radical, mono C₁-C₄Alkylamino radical C₁-C₄Alkyl, di-C₁-C₄Alkylamino radical C₁-C₄Alkyl, and the heteroaryl is a 5-12 membered monocyclic or polycyclic ring;

in a more preferred aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, including isomers thereof, wherein,

R¹selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl, wherein said heteroaryl may be independently selected from deuterium,Halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring;

R²is selected from heteroaryl, wherein said heteroaryl may be independently selected from halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring;

alternatively, the first and second electrodes may be,

R³selected from hydrogen, deuterium;

R⁴selected from the group consisting of heteroarylamino, heteroarylaminoaryl (said heteroaryl being, for example, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolylThiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl; such as phenyl), 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino, wherein the heteroarylamino, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

R³and R⁴Together with the nitrogen atom to which they are both attached form a 3-15 membered mono-or polycyclic heterocycle, wherein the 3-15 membered mono-or polycyclic heterocycle may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

R^aselected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, aryl, heteroaryl, heteroarylamino (said heteroaryl being for example furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl), wherein said aryl, heteroaryl may be substituted with one or more substituents independently selected from hydrogen, deuterium, cyano, nitro, such as benzoyl, pyridyl, and the like,C₁-C₄Alkyl, halo C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkoxy, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino radical, mono C₁-C₄Alkylamino radical C₁-C₄Alkyl, di-C₁-C₄Alkylamino radical C₁-C₄Alkyl, and the heteroaryl is a 5-12 membered monocyclic or polycyclic ring;

in another more preferred aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, including isomers thereof, wherein,

R¹selected from hydrogen, deuterium, C₁-C₄Alkyl, heteroaryl, wherein said heteroaryl may be independently selected from deuterium, halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring;

R²is selected from heteroaryl, wherein said heteroaryl may be independently selected from halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring;

alternatively, the first and second electrodes may be,

R¹and R²Together with the carbon atom to which they are commonly attached form a heteroaryl (e.g., dihydroquinolinyl) or 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from the group consisting of halogen, amino, cyano, C₁-C₄Alkyl, aryl, heteroaryl, and heteroaryl,Halogen substituted C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to aryl or heteroaryl, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle;

R³selected from hydrogen, deuterium;

R⁴selected from the group consisting of heteroarylaminoaryl (said heteroaryl being, for example, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl; said aryl being, for example, phenyl), 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino, wherein said 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

in another still more preferred aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, including isomers thereof, wherein,

R¹selected from hydrogen, deuterium, C₁-C₄An alkyl group;

R²selected from pyridyl, wherein the pyridyl may be independently selected from halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, mono C₁-C₄Alkylamino, di-C₁-C₄Radical substitution of alkylamino;

alternatively, the first and second electrodes may be,

R¹and R²Together with the carbon atom to which they are commonly attached form a heteroaryl group (e.g., dihydroquinolinyl) or a 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycleOptionally substituted by one or more groups selected from halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to aryl or heteroaryl, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle;

R³selected from hydrogen, deuterium;

R⁴selected from heteroarylaminoaryl (said heteroaryl being, for example, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl; said aryl being, for example, phenyl), 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

in a particularly preferred aspect, the present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, including isomers thereof, wherein,

R¹and R²Together with the carbon atom to which they are commonly attached form a heteroaryl (e.g., dihydroquinolinyl) or 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from the group consisting of halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄The radical substitution of an alkylamino radical,wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to an aryl or heteroaryl group, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle;

R³selected from hydrogen, deuterium;

alternatively, the first and second electrodes may be,

R^aselected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, aryl, heteroaryl, heteroarylamino (said heteroaryl being, for example, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolylThiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl), wherein said aryl, heteroaryl may be independently selected from one or more of hydrogen, deuterium, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkoxy, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino radical, mono C₁-C₄Alkylamino radical C₁-C₄Alkyl, di-C₁-C₄Alkylamino radical C₁-C₄Alkyl, and the heteroaryl is a 5-12 membered monocyclic or polycyclic ring;

in another particularly preferred aspect, the invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, including isomers thereof, wherein,

in a particularly preferred aspect, the invention relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, including isomers thereof, wherein,

R¹and R²Together with the carbon atom to which they are commonly attached form a heteroaryl (e.g., dihydroquinolinyl) or 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from the group consisting of halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, ammoniaRadical C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to aryl or heteroaryl, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle;

R^aselected from aryl, heteroaryl, heteroarylamino (said heteroaryl being for example furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl), wherein said aryl, heteroaryl may be substituted with one or more groups independently selected from hydrogen, deuterium, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino radical, mono C₁-C₄Alkylamino radical C₁-C₄Alkyl, di-C₁-C₄Alkylamino radical C₁-C₄Alkyl, and the heteroaryl is a 5-12 membered monocyclic or polycyclic ring.

The present invention relates to compounds of formula (I) or pharmaceutically acceptable salts thereof, including but not limited to:

definition of terms

The "halogen" in the present invention is fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

The "alkyl group" according to the present invention", includes straight or branched chain alkyl groups. C in the invention₁-C₄Alkyl means an alkyl group having 1 to 4 carbon atoms, preferably methyl, ethyl, propyl or isopropyl, n-butyl, isobutyl or tert-butyl. The alkyl groups in the compounds of the present invention may be optionally substituted or unsubstituted, and the substituents may include alkyl, halogen, alkoxy, haloalkyl, cyano, hydroxy, and the like. Examples of alkyl groups of the present invention include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and the like.

The "alkoxy group" as referred to herein means a group formed by linking the above alkyl group to an oxygen atom having free bonding ability, such as methoxy, ethoxy, propoxy, butoxy, isopropoxy, tert-butoxy, cyclopropoxy, etc.

The "alkylamino" in the present invention refers to a group formed by connecting the above alkyl group and amino group, such as methylamino, ethylamino, dimethylamino, etc.

The "halo C" of the present invention₁-C₄Alkyl "and" halo C₁-C₄Alkoxy "means an alkoxy group in which one or more hydrogen atoms are replaced by halogen atoms, in particular fluorine or chlorine atoms. In some embodiments, fluoro is preferred, e.g., CF₃、CHF₂、CH₂F、CH₂CH₂F、CH₂CHF₂、CH₂CF₃、OCF₃、OCHF₂、OCH₂F、OCH₂CH₂F、OCH₂CHF₂Or OCH₂CF₃。

The "spiro ring" in the present invention refers to a polycyclic hydrocarbon having a 3-15 membered monocyclic ring with one carbon atom (spiro atom) in common, wherein one or more double bonds may be contained, but none of the rings has a conjugated pi-electron system. The spirocycloalkyl group is classified into a single spirocycloalkyl group, a double spirocycloalkyl group or a multi spirocycloalkyl group according to the number of spiro atoms shared between rings, and preferably the single spirocycloalkyl group or the double spirocycloalkyl group.

The term "bridged ring" as used herein refers to a polycyclic hydrocarbon having more than two carbon atoms shared between 3-15 membered monocyclic rings, which may contain one or more double bonds, but none of the rings has a conjugated pi-electron system. And are classified into bicyclic hydrocarbons, tricyclic hydrocarbons, tetracyclic hydrocarbons, etc. according to the number of constituting rings.

"heterocyclic" as used herein refers to unsubstituted and substituted monocyclic or polycyclic non-aromatic ring systems, partially unsaturated or fully saturated ring systems containing one or more heteroatoms. Preferred heteroatoms include N, O and S. Monocyclic heterocycles include, but are not limited to, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, dihydroimidazolyl, dihydrofuranyl, piperidinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclic groups include spiro, bridged, fused, heterocyclic groups, which may be fused to aryl, heteroaryl, or cycloalkyl rings. Such as azaspiro [3.3] heptan-6-yl.

The "aryl" as used herein refers to a monocyclic or polycyclic aromatic ring system, such as a benzene ring, a naphthalene ring, and the like. The aryl group may be substituted or unsubstituted.

As used herein, "heteroaryl" refers to an aromatic ring system containing carbon and at least one heteroatom. Preferred heteroatoms include N, O and S. Heteroaryl groups can be monocyclic or polycyclic, substituted or unsubstituted. Monocyclic heteroaryl groups may have 1 to 4 heteroatoms in the ring, while polycyclic heteroaryl groups may contain 1 to 10 heteroatoms. The polycyclic heteroaryl ring may contain a fused spiro or bridged ring. Heteroaryl groups may be substituted or unsubstituted. Such as furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, or triazolyl; benzofuranyl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzotriazolyl, benzoxazolyl, furopyridinyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolyl, dihydroquinolinyl, naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl, pyridopyridinyl, pyrrolopyridinyl, quinolinyl, quinoxalinyl, quinazolinyl, thiadiazolopyrimidyl, and thienopyridinyl.

As used herein, "substituted" means that one or more hydrogen atoms in a group can be independently substituted with a substituent selected from the corresponding number of substituents. Those skilled in the art will be able to determine (experimentally or theoretically) possible or impossible substitution positions without undue effort.

In the compounds described herein, when a chemical name is specifically designated as (R) -or (S) -isomer, the main configuration should be understood as (R) -isomer or (S) -isomer, respectively. Any asymmetric carbon atom may be present in the (R) -, (S) -or (R, S) -configuration, preferably in the (R) -or (S) -configuration. The double bonds or especially the substituents on the ring can be present in cis (═ Z-) or trans (═ E-) or as cis-trans mixtures. The compounds described herein may thus be present as a mixture of isomers, or preferably as pure isomers, preferably as pure diastereomers or pure enantiomers.

The present invention relates to pharmaceutical compositions containing a compound of formula (I) as described above, including isomers, as an active ingredient.

The compounds of the present invention may optionally be used in combination with one or more other active ingredients, the respective amounts and proportions of which may be adjusted by the skilled person according to the particular condition and the particular circumstances of the patient, the clinical need, etc.

The compounds of formula (I) according to the invention, including isomers, can be prepared by a person skilled in the art (experience or reference).

The examples and preparations provided in this invention further illustrate and exemplify the compounds of this invention and their methods of preparation. It should be understood that the following preparation examples and examples do not limit the scope of the present invention in any way.

The following synthetic schemes describe the preparation of the compounds of formula (I) of the present invention, and the starting materials, reagents, catalysts, solvents, and the like, used in the following synthetic schemes can be prepared by methods well known to those of ordinary skill in the art of organic chemistry or are commercially available. All final derivatives of the invention can be prepared by methods described in the synthetic schemes or by methods analogous thereto, which are well known to those of ordinary skill in the art of organic chemistry. All variables used in these synthetic schemes are as defined below or according to the definition in the claims.

Preparation method

The following variables are defined as described above and the new variables are defined as described in this section. In addition, the compound shown in the general formula (I) and related intermediates can be purified by common separation methods, such as extraction, recrystallization, silica gel column chromatography separation and the like. The 200-mesh silica gel and the thin-layer chromatography silica gel plate are both produced by Qingdao ocean factories. The chemical reagents used were analytical or chemically pure commercial products of general reagents, and were used without further purification.

The invention provides a preparation method of a compound shown in a general formula (I), which is characterized by comprising the following steps:

the person skilled in the art knows how to obtain the compounds of the formula (I-c).

For example, the compound of formula (I-c) can be obtained by Buchwald-Hartwig coupling under appropriate basic conditions and in a solvent and in the presence of a catalyst which is a palladium reagent including, but not limited to, palladium acetate (Pd (OAc)₂) 1,1 '-binaphthyl-2, 2' -bis (diphenylphosphine) (BINAP), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (SPhos), dicyclohexyl (3, 6-dimethoxy-2 ',4',6 '-triisopropyl (1,1' -biphenyl) -2-yl) phosphine (Brettphos), tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (PdCl)₂(dppf)), [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride dichloromethane complex (PdCl)₂(dppf)·CH₂Cl₂) Tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Bis (tricyclohexylphosphine) palladium dichloride (PdCl)₂(P(Cy)₃)₂) (ii) a The basic reagent includes organic and inorganic bases including but not limited to Triethylamine (TEA), N-Diisopropylethylamine (DIPEA), N-butyllithium, lithium diisopropylamide, lithium bistrimethylsilyl amide, potassium acetate (KOAc), sodium t-butoxide (tBuONa), potassium t-butoxide (tBuOK), sodium hydride (NaH), potassium phosphate (K)₃PO₄) Sodium carbonate (Na)₂CO₃) Potassium carbonate (K)₂CO₃) Lithium hydroxide (KOH), sodium hydroxide (NaOH); the solvent includes, but is not limited to, Toluene (Toluene), 1,4-dioxane (1,4-dioxane), Tetrahydrofuran (THF), acetonitrile (CH)₃CN), N' -Dimethylformamide (DMF), and mixed solvents of these solvents in different proportions with water;

alternatively, the first and second electrodes may be,

for example, the compound of formula (I-c) can be obtained by nucleophilic substitution reaction under appropriate basic conditions and solvents, and reagents of basic conditions include organic and inorganic bases, including but not limited to Triethylamine (TEA), N-Diisopropylethylamine (DIPEA), N-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, potassium acetate (KOAc), sodium tert-butoxide (tBuONa), potassium tert-butoxide (tBuOK), sodium hydride (NaH), potassium phosphate (K)₃PO₄) Sodium carbonate (Na)₂CO₃) Potassium carbonate (K)₂CO₃) Lithium hydroxide (KOH), sodium hydroxide (NaOH); the solvent includes, but is not limited to, N' -Dimethylformamide (DMF), N-methylpyrrolidone (NMP), 1,4-dioxane (1,4-dioxane), Tetrahydrofuran (THF), acetonitrile (CH)₃CN)；

Carrying out one-pot reaction on the compound shown in the formula (I-c) to obtain a compound shown in the formula (I-d), wherein the deprotection is carried out by using an acid or alkali reagent under the condition of a proper solvent in the 1 st stage; the 2 nd stage is to react with N, N' -Thiocarbonyldiimidazole (TCDI) under the catalysis of a proper base reagent under the condition of a proper solvent to obtain the compound shown as the formula (I-d), wherein the solvent comprises Dichloromethane (DCM), 1,4-dioxane (1,4-dioxane) Tetrahydrofuran (THF), acetonitrile (CH)₃CN), N' -Dimethylformamide (DMF); the acid reagents include, but are not limited to, trifluoroacetic acid (TFA) and hydrochloric acid (HCl); the alkaline agent includes, but is not limited toPiperidine, diethylamine;

reacting the compound shown as the formula (I-d) with hydrazine hydrate under the condition of a proper solvent to obtain the compound shown as the formula (I-e), wherein the solvent comprises but is not limited to methanol (CH)₃OH), ethanol (EtOH);

reacting the compound shown as the formula (I-e) with the compound shown as the formula (I-f) under the condition of a proper solvent to obtain the compound shown as the formula (I), wherein the solvent comprises but is not limited to methanol (CH)₃OH), ethanol (EtOH);

P¹is a protecting group selected from tert-butyloxycarbonyl (Boc) or 9-fluorenylmethyloxycarbonyl (Fmoc).

The invention provides a novel thiohydrazide p53 protein activity regulator, which is used for treating or preventing tumors or cancers related to the target. Meanwhile, these compounds or pharmaceutical compositions comprising the same as an active ingredient, etc. can maximize the clinical efficacy against these diseases within a safe therapeutic window.

Detailed Description

Representative compounds of the present invention are prepared by the above-described illustrative methods, and the present invention is further described below with reference to the following examples, which are not intended to limit the scope of the present invention.

LC-MS analysis method:

mass spectrum conditions: instrument Thermo MSQ Plus; ion source ESI (EA + EA-); the voltage of the taper hole is 30V; capillary voltage 3.00 KV; the source temperature is 350 ℃;

chromatographic conditions are as follows: instrument Thermo U3000; detector DAD-3000(RS) (diode array detector); the chromatographic column is Shimadzu inert silica ODS-HL HP 3 mu m 3.0 multiplied by 100 mm; the flow rate is 0.4 mL/min; the column temperature is 30 ℃; mobile phase CH₃OH/H₂O/HCOOH(75/25/0.2)。

HPLC analytical method (a):

instrument Thermo U3000; detector VWD-3 × 00(RS) (ultraviolet detector); shimadzu Shim-pack VP-ODS 5 μm 4.6X 150mm column; the flow rate is 1.0 mL/min; the column temperature is 30 ℃; mobile phase ACH₃OH/H₂O(85/15)。

HPLC analytical method (ii):

instrument Thermo U3000; detector VWD-3X 00(RS) (ultraviolet detector); shimadzu Shim-pack VP-ODS 5 μm 4.6X 150mm column; the flow rate is 1.0 mL/min; the column temperature is 30 ℃; mobile phase BCH₃OH/H₂O/TEA/HCOOH(65/35/0.1/0.2)。

HPLC analytical method (iii):

instrument Thermo U3000; detector VWD-3 × 00(RS) (ultraviolet detector); shimadzu Shim-pack VP-ODS 5 μm 4.6X 150mm column; the flow rate is 1.0 mL/min; the column temperature is 30 ℃; mobile phase CCH₃OH/H₂O(75/25)。

HPLC analytical method (d):

instrument Thermo U3000; detector VWD-3 × 00(RS) (ultraviolet detector); shimadzu Shim-pack VP-ODS 5 μm 4.6X 150mm column; the flow rate is 1.0 mL/min; the column temperature is 30 ℃; mobile phase DCH₃OH/H₂O(65/35)。

HPLC analytical method (v):

instrument Thermo U3000; detector VWD-3 × 00(RS) (ultraviolet detector); shimadzu Shim-pack VP-ODS 5 μm 4.6X 150mm column; the flow rate is 1.0 mL/min; the column temperature is 30 ℃; mobile phase ECH₃OH/H₂O(80/20)。

HPLC analytical method (vi):

instrument Thermo U3000; detector VWD-3 × 00(RS) (ultraviolet detector); shimadzu Shim-pack VP-ODS 5 μm 4.6X 150mm column; the flow rate is 1.0 mL/min; the column temperature is 30 ℃; mobile phase FCH₃OH/H₂O(70/30)。

¹H-NMR analysis method:

¹H-NMR in DMSO-d at room temperature using a BRUKERAVANCE-400MHz NMR spectrometer₆Or CDCl₃In the above, the signal peaks are represented by s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet) and dd (doublet) when measured using TMS as an internal standard. Coupling constants (J) are in Hertz (Hz).

Abbreviations:

1, 4-dioxane; DCM dichloromethane

PE petroleum ether; EA Ethyl acetate

Toluene Toluene; n-Hexane

CH₃OH methanol; EtOH ethanol

DMF N, N' -dimethylformamide; TEA Triethylamine

TFA trifluoroacetic acid; HCl hydrochloric acid

Pyridine; na (Na)₂SO₄Sodium sulfate

TCDI N, N' -thiocarbonyldiimidazole; tf₂O-Trifluoromethanesulfonic anhydride

K₂CO₃Potassium carbonate; CS₂CO₃Cesium carbonate

tBuONa sodium tert-butoxide; tBuOK Potassium tert-butoxide

N₂H₄·H₂Hydrazine hydrate; pd (OAc)₂Palladium acetate

BINAP 1,1 '-binaphthyl-2, 2' -bisdiphenylphosphine; pd₂(dba)₃Tris (dibenzylideneacetone) dipalladium

Sphos 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl

Brettphos dicyclohexyl (3, 6-dimethoxy-2 ',4',6 '-triisopropyl (1,1' -biphenyl) -2-yl) phosphine

TLC thin layer chromatography

Representative compounds I-1 to I-25 (see Table 1) were prepared according to the above-described procedure.

TABLE 1 representative Compounds I-1 to I-25 of the invention

The present invention will be further described with reference to specific examples, but the scope of the present invention is not limited to these examples. The percentages stated in the present invention are percentages by weight, unless otherwise indicated. The numerical ranges set forth in this specification, such as units of measure, reaction conditions, physical states of compounds, or percentages, are intended to provide an unambiguous written reference thereto. One skilled in the art, in practicing the invention, may still obtain desirable results using temperatures, concentrations, amounts, numbers of carbon atoms, etc., outside of these ranges or other than the individual values.

Example 1

N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -9- (pyridin-2-yl) -3, 9-diazaspiro [5.5] undecane-3-thiohydrazide I-1

Intermediate: preparation of tert-butyl 9- (pyridin-2-yl) -3, 9-diazaspiro [5.5] undecane-3-carboxylate I-1c (first step)

Mixing commercially available I-1a (316.0mg,2.0mmol,1.0eq) and I-1b (508.0mg,2.0mmol,1.0eq), Pd₂(dba)₃(183.0mg,0.2mmol,0.1eq), BINAP (186.6mg,0.3mmol,0.15eq), tBuOK (672.0mg,6.0mmol,3.0eq), TEA (606.0mg,6.0mmol,3.0eq), and Toluene (20mL) in N₂Heating to 110 ℃ under protection and stirring for 8h, and cooling to ambient temperature. Concentrating the reaction solution, separating a crude product by silica gel column chromatography (EA/n-Hexane (v/v) ═ 1/4-1/2 elution), and concentrating to obtain a yellow solid I-1 c. (367.0mg, yield 55.4%). LC-MS MS-ESI (M/z)332.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (9- (pyridin-2-yl) -3, 9-diazaspiro [5.5] undecan-3-yl) thione I-1d (second step)

I-1c (367.0mg,1.1mmol,1.0eq) was dissolved in DCM (5mL), TFA (5mL) was added, stirred at ambient temperature for 1h, and concentrated to give the trifluoroacetate salt of secondary amine as a brown oil, which was used in the next stage reaction without further treatment. The trifluoroacetate salt of the secondary amine was dissolved in DCM (10mL) and TEA (C) (0) was added333.3mg,3.3mmol,3.0eq) and TCDI (213.8mg,1.2mmol,1.1eq) was stirred at ambient temperature for 16 h. The reaction was diluted with DCM (200mL), washed 2 times with water and Na anhydrous₂SO₄The organic phase was dried and concentrated. The crude product is subjected to preparative TLC (CH)₂Cl₂/CH₃OH (v/v) ═ 12/1) was isolated to give I-1d as a yellow solid. (281.0mg, yield 74.9%). LC-MS MS MS-ESI (M/z)342.2[ M + H]⁺。

Intermediate: preparation of 9- ((pyridin-2-yl) -3, 9-diazaspiro [5.5] undecane-3-thiohydrazide I-1e (third step)

Intermediate I-1d (281.0mg,0.82mmol,1.0eq) was dissolved in EtOH (15mL), hydrazine hydrate (61.5mg,1.23mmol,1.5eq) was added at a concentration of 80%, and the resulting solution was heated to 95 ℃ and stirred for 2h, cooled to ambient temperature. The reaction solution was concentrated to give crude I-1e as a yellow solid which was used directly in the next reaction. (by 100%). LC-MS MS-ESI (M/z)306.2[ M + H]⁺。

A compound: preparation of N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -9- (pyridin-2-yl) -3, 9-diazaspiro [5.5] undecane-3-thiohydrazide I-1 (fourth step)

Intermediate I-1e (0.82mmol,1.0eq) was dissolved in EtOH (15mL), commercial I-1f (132.5mg,0.90mmol,1.1eq) was added and the resulting solution heated to 95 ℃ and stirred for 8h, cooled to ambient temperature. The solid was collected by filtration and successively treated with CH₃OH (20mL) and EtOAc (20mL) were each washed 1 time in syrup to give I-1 as a yellow solid. (107mg, yield 30.1%). LC-MS MS-ESI (M/z)435.2[ M + H]⁺. Compound I-1 is a mixture of formula E and formula Z, with the Z/E being about 1: 0.18.¹H-NMR(400MHz,CDCl₃)δppm 14.77(s,1H),8.59(d,J＝3.6Hz,1H),8.18(d,J＝3.2Hz,1H),7.44-7.51(m,2H),7.19-7.26(m,1H),6.66(d,J＝8.4Hz,1H),6.58(t,J＝6.0Hz,1H),4.11(s,4H),3.55(s,4H),3.08(t,J＝6.0Hz,2H),2.87(t,J＝6.0Hz,2H),2.04(t,J＝6.0Hz,2H),1.63-1.66(m,8H)。

Example 2

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -1, 4-diazepan-1-thiohydrazide I-2

Intermediate: preparation of 4- (pyridin-2-yl) -1, 4-diazepan-1-carboxylic acid tert-butyl ester I-2c

A yellow solid I-2c was prepared from 2-bromopyridine I-1a (474.0mg,3.0mmol,1.0eq) and 1, 4-diazepane-1-carboxylic acid tert-butyl ester I-2b (601.0mg,3.0mmol,1.0eq) following the first analogous procedure in example 1. (465.0mg, 55.9% yield). LC-MS MS-ESI (M/z)278.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (4- (pyridin-2-yl) -1, 4-diazepan-1-yl) methinone I-2d

A yellow solid, I-2d, was prepared from I-2c (465.0mg,1.68mmol,1.0eq), TFA (5mL), TEA (509.0mg,5.04mmol,3.0eq) and TCDI (329.7mg,1.85mmol,1.1eq) following the second analogous procedure in example 1. (246.0mg, yield 51.0%). LC-MS MS-ESI (M/z)288.2[ M + H]⁺。

Intermediate: preparation of 4- (pyridin-2-yl) -1, 4-diazepan-1-thiohydrazide I-2e

A yellow solid, I-2e, was prepared from I-2d (246.0mg,0.86mmol,1.0eq) and 80% hydrazine hydrate (80.6mg,1.3mmol,1.5eq) by a procedure similar to the third step in example 1. (by 100%). LC-MS MS-ESI (M/z)252.1[ M + H]⁺。

A compound: (E) preparation of (E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -1, 4-diazepan-1-thiohydrazide I-2

A yellow solid I-2 was prepared from I-2e (0.86mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (139.8mg,0.95mmol,1.1eq) following a 4 th step analogous to that in example 1. The crude product was subjected to preparative TLC (DCM/CH)₃OH (v/v) ═ 12/1) was separated, and then concentrated. (76.0mg, yield 23.3%). LC-MS MS-ESI (M/z)381.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm8.14(d,J＝3.6Hz,1H),7.69(s,1H),7.34-7.43(m,2H),6.89(s,1H),6.49-6.53(m,2H),4.12-4.22(m,2H),3.80(d,J＝6.0Hz,4H),3.61(s,2H),3.16(t,J＝6.0Hz,2H),2.80(t,J＝5.6Hz,2H),1.95-2.18(m,4H)。

Example 3

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -7- (pyridin-2-yl) -2, 7-diazaspiro [4.4] nonane-2-thiohydrazide I-3

Intermediate: preparation of 7- (pyridin-2-yl) -2, 7-diazaspiro [4.4] nonane-2-carboxylic acid tert-butyl ester I-3c

Yellow solid I-3c is prepared from 2-bromopyridine I-1a (316.0mg,2.0mmol,1.0eq) and 2, 7-diazaspiro [4.4]]Nonane-2-carboxylic acid tert-butyl ester I-3b (452.6mg,2.0mmol,1.0eq) was prepared by analogy with the first step in example 1. (417.0mg, yield 68.8%). LC-MS MS MS-ESI (M/z)304.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (7- (pyridin-2-yl) -2, 7-diazaspiro [4.4] nonan-2-yl) thione I-3d

A yellow solid I-3d was prepared from I-3c (417.0mg,1.38mmol,1.0eq), TFA (5mL), TEA (418.1mg,4.14mmol,3.0eq) and TCDI (270.9mg,1.52mmol,1.1eq) following the second analogous procedure in example 1. (212.0mg, yield 49.1%). LC-MS MS-ESI (M/z)314.1[ M + H]⁺。

Intermediate: preparation of 7- (pyridin-2-yl) -2, 7-diazaspiro [4.4] nonane-2-thiohydrazide I-3e

A yellow solid I-3e was prepared from I-3d (212.0mg,0.67mmol,1.0eq) and 80% hydrazine hydrate (62.7mg,1.0mmol,1.5eq) by a similar procedure as the third step in example 1. (by 100%). LC-MS MS-ESI (M/z)278.1[ M + H]⁺。

A compound: (E) preparation of (E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -7- (pyridin-2-yl) -2, 7-diazaspiro [4.4] nonane-2-thiohydrazide I-3

A yellow solid I-3 was prepared from I-3e (0.67mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (108.9mg,0.74mmol,1.1eq) following a 4 th step analogous to that in example 1. Subjecting the crude product to reverse phase preparative chromatography (C18 column, EZ Plus 100D full-automatic rapid medium pressure preparative chromatography system with phase A of water and phase B of MeOH (5% -100%), retention time of 50min, phase B of AThe peak of the target product appears when the alcohol content is 89%), and then the product is concentrated. (115mg, yield 42.3%). LC-MS MS-ESI (M/z)407.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.14(s,1H),7.78(d,J＝4.0Hz,1H),7.43(s,1H),7.35(d,J＝7.2Hz,1H),6.94(t,J＝6.4Hz,1H),6.52(d,J＝5.2Hz,1H),6.33(s,1H),3.75-3.87(m,4H),3.40-3.59(m,4H),3.13(s,2H),2.81(s,2H),1.97-2.07(m,6H)。

Example 4

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -7- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide I-4

Intermediate: preparation of 7- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester I-4c

Yellow solid I-4c is prepared from 2-bromopyridine I-1a (1.58g,10.0mmol,1.0eq) and 2, 7-diazaspiro [3.5]]Nonane-2-carboxylic acid tert-butyl ester I-4b (2.26g,10.0mmol,1.0eq) was prepared by analogy with the first step in example 1. (434.0mg, yield 14.3%). LC-MS MS-ESI (M/z)303.2[ M + H ]]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (7- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonan-2-yl) methylthioone I-4d

A yellow solid I-4d was prepared from I-4c (434.0mg,1.43mmol,1.0eq), TFA (5mL), TEA (433.3mg,4.29mmol,3.0eq) and TCDI (279.8mg,1.57mmol,1.1eq) following the second analogous procedure in example 1. (177.0mg, yield 39.5%). LC-MS MS-ESI (M/z)313.1[ M + H]⁺。

Intermediate: preparation of 7- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide I-4e

A yellow solid I-4e was prepared from I-4d (177.0mg,0.56mmol,1.0eq) and 80% hydrazine hydrate (52.5mg,0.84mmol,1.5eq) by following the third similar procedure as in example 1. (by 100%). LC-MS MS-ESI (M/z)278.1[ M + H]⁺。

A compound: (E) preparation of (E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -7- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide I-4

I-4 was prepared from I-4e (0.56mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (91.3mg,0.62mmol,1.1eq) by analogy with step 4 of example 1. The mixture was subjected to preparative TLC (DCM/CH) 2 more times₃OH (v/v) ═ 12/1) was separated and concentrated. (46mg, yield 20.2%). LC-MS MS-ESI (M/z)407.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.17-8.18(m,1H),7.80(d,J＝4.4Hz,1H),7.43-7.48(m,1H),7.38(d,J＝7.6Hz,1H),6.97(dd,J＝5.2Hz,J＝7.6Hz,1H),6.67(d,J＝8.4Hz,1H),6.58(dd,J＝5.2Hz,J＝7.2Hz,1H),3.89-3.99(m,4H),3.53(s,4H),3.14-3.20(m,2H),2.82(s,2H),1.99(s,2H),1.85(s,4H)。

Example 5

(3aR,6aS) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -thiohydrazide I-5

Intermediate: preparation of (3aR,6aS) -5- (pyridin-2-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -carboxylic acid tert-butyl ester I-5c

Yellow solid I-5c was prepared from 2-bromopyridine I-1a (316.0mg,2.0mmol,1.0eq) and (3aR,6aS) -hexahydropyrrolo [3,4-c ]]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester I-5b (424.6mg,2.0mmol,1.0eq) was prepared by analogy with the first step in example 1. (390.0mg, yield 67.5%). LC-MS MS-ESI (M/z)290.2[ M + H [ ]]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) ((3aR,6aS) -5- (pyridin-2-yl) hexahydropyrrolo [3,4-c ] pyrrol-2 (1H) -yl) methinone I-5d

A yellow solid I-5d was prepared from I-5c (390.0mg,1.35mmol,1.0eq), TFA (5mL), TEA (409.1mg,4.05mmol,3.0eq) and TCDI (263.7mg,1.48mmol,1.1eq) following the second analogous procedure in example 1. (108.0mg, yield 26.7%). LC-MS MS MS-ESI (M/z)300.1[ M + H]⁺。

Intermediate: preparation of (3aR,6aS) -5- (pyridin-2-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -thiohydrazide I-5e

A yellow solid I-5e was prepared from I-5d (108.0mg,0.36mmol,1.0eq) and 80% hydrazine hydrate (33.8mg,0.54mmol,1.5eq) by a similar procedure as the third step in example 1. (by 100%). LC-MS MS-ESI (M/z)263.1[ M + H]⁺。

A compound: preparation of (3aR,6aS) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -thiohydrazide I-5

I-5 was prepared from I-5e (0.36mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (58.9mg,0.40mmol,1.1eq) following a procedure similar to that of example 1, the crude product was isolated by reverse phase preparative chromatography (C18 column, EZ Plus 100D full automatic flash medium pressure preparative chromatography system, phase A: water, phase B: MeOH (5% -100%), retention time 50min, 85% MeOH in phase B) and concentrated. (27.2mg, yield 19.2%). LC-MS MS-ESI (M/z)393.2[ M + H ]]⁺. Compound I-5 is a mixture of formula E and formula Z, with E/Z being about 1: 1.¹H-NMR(400MHz,CDCl₃)δppm 14.58(s,1H),8.58(s,1H),8.15(s,1H),7.56-7.36(m,2H),7.22(d,J＝4.8Hz,1H),6.53-6.59(m,2H),6.35(t,J＝7.8Hz,1H).，4.01-4.23(m,3H),3.49-3.75(m,3H),3.18(s,2H),3.06-3.08(m,3H),2.87-2.91(m,3H),2.02(d,J＝5.6Hz,2H)。

Example 6

(E) -2- (6, 7-dihydroquinolin-8 (5H) -ylidene) -N- (4- (pyridin-2-ylamino) phenyl) hydrazine-1-thioamide I-6

Intermediate: preparation of tert-butyl (4- (pyridin-2-ylamino) phenyl) carbamate I-6c

A yellow solid I-6c was prepared from 2-bromopyridine I-1a (474.0mg,3.0mmol,1.0eq) and tert-butyl (4-aminophenyl) carbamate I-6b (624.0mg,3.0mmol,1.0eq) following the first analogous procedure in example 1. (374.0mg, yield 65.6%). LC-MS MS-ESI (M/z)286.2[ M + H]⁺。

Intermediate: preparation of N- (4- (pyridin-2-ylamino) phenyl) -1H-imidazole-1-thioamide I-6d

A yellow solid, I-6d, was prepared from I-6c (374.0mg,1.31mmol,1.0eq), TFA (5mL), TEA (396.9mg,3.93mmol,3.0eq), and TCDI (256.6mg,1.44mmol,1.1eq) following the second analogous procedure in example 1. (134.0mg, yield 34.4%). LC-MS MS-ESI (M/z)296.1[ M + H]⁺。

Intermediate: preparation of N- (4-pyridin-2-ylamino) phenyl) hydrazine thioamide I-6e

A yellow solid I-6e was prepared from I-6d (134.0mg,0.45mmol,1.0eq) and 80% hydrazine hydrate (42.1mg,0.68mmol,1.5eq) by the third similar procedure as in example 1. (by 100%). LC-MS MS-ESI (M/z)260.1[ M + H]⁺。

A compound: (E) preparation of (E) -2- (6, 7-dihydroquinolin-8 (5H) -ylidene) -N- (4- (pyridin-2-ylamino) phenyl) hydrazine-1-thioamide I-6

I-6 was prepared from intermediate I-6e (0.45mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (72.1mg,0.49mmol,1.1eq) following a similar procedure as in compound I-1. (51.3mg, yield 29.4%). LC-MS MS-ESI (M/z)389.2[ M + H]⁺。¹H-NMR(400MHz,DMSO-d₆)δppm 9.13(s,1H),8.82(s,1H),8.12(d,J＝4.0Hz,1H),7.73-7.77(m,3H),7.67(d,J＝7.6Hz,1H),7.49-7.55(m,3H),7.24(dd,J＝5.2Hz,J＝7.6Hz,1H),6.77(d,J＝8.4Hz,1H),6.67(t,J＝6.0Hz,1H),3.19(d,J＝6.0Hz,2H),2.88(d,J＝5.2Hz,2H),1.94-2.02(m,2H)。

Example 7

N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) -1, 5-diazacyclooctane-1-thiohydrazide I-7

Intermediate: preparation of 5- (pyridin-2-yl) -1, 5-diazacyclooctane-1-carboxylic acid tert-butyl ester I-7c

Yellow solid I-7c was prepared from 2-bromopyridine I-1a (316.0mg,2.0mmol,1.0eq) and 1, 5-diazacyclooctane-1-carboxylic acid tert-butyl ester I-7b (428.0mg,2.0mmol,1.0eq) as described in example 1Prepared by one step of similar steps. (226.0mg, yield 38.8%). LC-MS MS-ESI (M/z)292.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (5- (pyridin-2-yl) -1, 5-diazacyclooctan-1-yl) methylsulfan I-7d

A yellow solid I-7d was prepared from I-7c (226.0mg,0.78mmol,1.0eq), TFA (5mL), TEA (236.3mg,2.34mmol,3.0eq) and TCDI (153.3mg,0.86mmol,1.1eq) following the second analogous procedure in example 1. (76.0mg, yield 32.4%). LC-MS MS-ESI (M/z)302.1[ M + H]⁺。

Intermediate: preparation of 5- (pyridin-2-yl) -1, 5-diazacyclooctane-1-thiohydrazide I-7e

A yellow solid I-7e was prepared from I-7d (76.0mg,0.25mmol,1.0eq) and 80% hydrazine hydrate (31.3mg,0.37mmol,1.5eq) by a similar procedure as the third step in example 1. (by 100%). LC-MS MS-ESI (M/z)266.1[ M + H ].

A compound: preparation of N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) -1, 5-diazacyclooctane-1-thiohydrazide I-7

I-7 was prepared from I-7e (0.25mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (39.7mg,0.27mmol,1.1eq) following a similar procedure as in compound I-4. (13.5mg, yield 26.4%). LC-MS MS-ESI (M/z)395.2[ M + H]⁺. Compound I-5 is a mixture of formula E and formula Z, with a Z/E of about 1: 0.7.¹H-NMR(400MHz,CDCl₃)δppm 14.71(s,1H),8.57(d,J＝3.6Hz,1H),8.15(d,J＝4.0Hz,1H),7.50(d,J＝7.6Hz,1H),7.46-7.34(m,1H),7.20(dd,J＝7.6,4.4Hz,1H),6.55-6.47(m,2H),4.15(s,2H),3.63(s,2H),3.49(s,2H),3.07(t,J＝6.4Hz,2H),2.87(dd,J＝11.0,4.9Hz,2H),2.26(s,6H),2.05(d,J＝5.8Hz,2H)。

Example 8

(E) -7- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide I-8

Intermediate: preparation of 7- (4-cyanopyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-2-carboxylic acid tert-butyl ester I-8c

Commercial 4-cyano-2-fluoropyridine I-8a (122.0mg,1.0mmol,1.0eq) was dissolved in DMF (15mL) and 2, 7-diazaspiro [3.5] was added]Nonane-2-carboxylic acid tert-butyl ester I-4b (226.0mg,1.0mmol,1.0eq) and K₂CO₃(414.0mg,3.0mmol,3.0 eq). The resulting mixture was heated to 110 ℃ and stirred for 3h, cooled to ambient temperature. The reaction was quenched with water (100mL) and extracted 3 times with EA (50 mL). The combined organic phases were washed 3 times with saturated brine (100mL) anhydrous Na₂SO₄And (5) drying. Concentrating the reaction solution, separating a crude product by silica gel column chromatography (EA/n-Hexane (v/v) ═ 1/4-1/2 elution), and concentrating to obtain an off-white solid I-8 c. (317.0mg, yield 96.6%). LC-MS MS MS-ESI (M/z)329.2[ M + H ]]⁺。

Intermediate: preparation of 2- (2- (1H-imidazole-1-thiocarbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) isonicotinonitrile I-8d

A yellow solid I-8d was prepared from intermediate I-8c (317.0mg,0.97mmol,1.0eq), TFA (5mL), TEA (293.9mg,2.91mmol,3.0eq) and TCDI (190.7mg,1.07mmol,1.1eq) following the second analogous procedure in example 1. (176.0mg, yield 53.7%). LC-MS MS-ESI (M/z)339.1[ M + H]⁺。

Intermediate: preparation of 7- (4-cyanopyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide I-8e

A yellow solid I-8e was prepared from I-8d (176.0mg,0.52mmol,1.0eq) and 80% hydrazine hydrate (48.8mg,0.78mmol,1.5eq) by a similar procedure as the third step in example 1. (by 100%). LC-MS MS-ESI (M/z)303.1[ M + H]⁺。

A compound: (E) preparation of (E) -7- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide I-8

I-8 was prepared from I-8e (0.52mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (83.9mg,0.57mmol,1.1eq) following a similar procedure for compound I-3. (37.0mg, yield 16.5%). LC-MS MS-ESI (M/z)432.2[ M + H [ ]]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.25(d,J＝5.0Hz,1H),7.79(d,J＝4.6Hz,1H),7.39(d,J＝7.6Hz,1H),6.97(dd,J＝7.3,5.1Hz,1H),6.83(s,1H),6.71(d,J＝4.9Hz,1H),3.90-3.99(m,4H),3.58(s,4H),3.05-3.27(m,2H),2.83(s,2H),1.99(s,2H),1.85(s,4H)。

Example 9

(E) -4- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -1, 4-diazepan-1-thiohydrazide I-9

Intermediate: preparation of 4- (4-cyanopyridin-2-yl) -1, 4-diazepan-1-carboxylic acid tert-butyl ester I-9c

A white solid I-9c was prepared from 4-cyano-2-fluoropyridine I-8a (366.0mg,3.0mmol,1.0eq) and 1, 4-diazepane-1-carboxylic acid tert-butyl ester I-2b (601.0mg,3.0mmol,1.0eq) following a similar procedure for intermediate I-8 c. (893.0mg, yield 98.6%). LC-MS MS-ESI (M/z)303.2[ M + H ]]⁺。

Intermediate: preparation of 2- (4- (1H-1-thiocarbonyl) -1, 4-diazepan-1-yl) isonicotinonitrile I-9d

A yellow solid I-9d was prepared from intermediate I-9c (302.0mg,1.0mmol,1.0eq), TFA (5mL), TEA (303.0mg,3.0mmol,3.0eq), and TCDI (196.0mg,1.1mmol,1.1eq) following the second analogous procedure in example 1. (212.0mg, yield 67.7%). LC-MS MS-ESI (M/z)313.1[ M + H]⁺。

Intermediate: preparation of 4- (4-cyanopyridin-2-yl) -1, 4-diazepan-1-thiohydrazide I-9e

A yellow solid I-9e was prepared from intermediate I-9d (212.0mg,0.68mmol,1.0eq) and 80% hydrazine hydrate (63.7mg,1.02mmol,1.5eq) following the third analogous procedure in example 1. (by 100%). LC-MS MS-ESI (M/z)277.1[ M + H ]]⁺。

A compound: (E) preparation of (E) -4- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -1, 4-diazepan-1-thiohydrazide I-9

I-9 is composed of I-9e (0.68mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (110.4mg,0.75mmol,1.1eq) was prepared following a similar procedure as in compound I-3. (72.0mg, yield 26.1%). LC-MS MS-ESI (M/z)406.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.23(d,J＝5.0Hz,1H),7.66(s,1H),7.37(d,J＝7.5Hz,1H),6.82-6.94(m,1H),6.59-6.77(m,2H),4.14(s,2H),3.40-4.03(m,6H),3.16(t,J＝6.2Hz,2H),2.81(t,J＝10.8Hz,2H),1.86-2.24(m,4H)。

Example 10

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (quinolin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-10

Intermediate: preparation of 6- (quinolin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylic acid tert-butyl ester I-10c

Quinolin-2-yl trifluoromethanesulfonate I-10a (636.0mg,2.3mmol,1.0eq), 2, 6-diazaspiro [3.3]]Heptane-2-carboxylic acid tert-butyl hemioxalate I-10b (662.4mg,2.3mmol,1.0eq), Cs₂CO₃(1.50g,4.6mmol,2.0eq)，Pd(OAc)₂(51.5mg,0.23mmol,0.1eq), BINAP (214.6mg,0.35mmol,0.15eq) and Toluene (20mL) were placed in a sealed tube, purged with nitrogen for 5min and sealed quickly, heated to 100 ℃ and stirred for 8h, cooled to ambient temperature. The reaction solution was concentrated, and the crude product was separated by silica gel column chromatography (EA/n-Hexane (v/v) ═ 1/2-1/1) to give a white solid I-10 c. (617.0mg, yield 82.5%). LC-MS MS-ESI (M/z)326.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (6- (quinolin-2-yl) -2, 6-diazaspiro [3.3] heptan-2-yl) methinone I-10d

A yellow solid I-10d was prepared from intermediate I-10c (617.0mg,1.9mmol,1.0eq), TFA (10mL), TEA (575.7mg,5.7mmol,3.0eq) and TCDI (374.2mg,2.1mmol,1.1eq) following the second analogous procedure in example 1. (412.0mg, yield 64.7%). LC-MS MS-ESI (M/z)336.1[ M + H]⁺。

Intermediate: preparation of 6- (quinolin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-10e

A yellow solid I-10e was prepared from intermediate I-10d (412.0mg,1.23mmol,1.0eq) and 80% hydrazine hydrate (115.4mg,1.85mmol,1.5eq) following the third analogous procedure in example 1. (by 100%). LC-MS MS MS-ESI (M/z)300.1[ M + H]⁺。

A compound: (E) preparation of (E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (quinolin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-10

I-10 was prepared from I-10e (1.23mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (198.7mg,1.35mmol,1.1eq) following a similar procedure as in compound I-4. (26.0mg, yield 4.9%). LC-MS MS-ESI (M/z)429.2[ M + H ]]⁺。¹H-NMR(400MHz,CDCl₃)δppm 7.88(d,J＝8.8Hz,1H),7.81(d,J＝4.2Hz,1H),7.77(d,J＝7.9Hz,1H),7.61(d,J＝7.9Hz,1H),7.55(t,J＝7.6Hz,1H),7.41(d,J＝7.5Hz,1H),7.23(d,J＝7.3Hz,1H),6.96-7.05(m,1H),6.59(d,J＝8.8Hz,1H),4.43(s,2H),4.34(s,6H),3.18-3.19(m,2H),2.84(s,2H),2.03(d,J＝15.2Hz,2H)。

Example 11

(E) -6- (4-cyanopyridin-2-yl) -N' - (1- (pyridin-2-yl) ethylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-11

Intermediate: preparation of 6- (4-cyanopyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylic acid tert-butyl ester I-11c

White solid I-11c was prepared from 4-cyano-2-fluoropyridine I-8a (610.0mg,5.0mmol,1.0eq) and 2, 6-diazaspiro [3.3]]Heptane-2-carboxylic acid tert-butyl hemioxalate I-10b (1.44g,5.0mmol,1.0eq) was prepared by a similar procedure as in intermediate I-8 c. (1.34g, yield 89.4%). LC-MS MS-ESI (M/z)301.2[ M + H [ ]]⁺。

Intermediate: preparation of 2- (6- (1H-imidazole-1-thiocarbonyl) -2, 6-diazaspiro [3.3] heptan-2-yl) isonicotinonitrile I-11d

Yellow solid I-11d was prepared from intermediate I-11c (1.34g,4.47mmol,1.0eq), TFA (10mL), TEA (1.35g,13.41mmol,3.0eq) and TCDI (876.7mg,4.92mmol,1.1eq) were prepared by following the second analogous procedure in example 1. (199.0mg, yield 14.4%). LC-MS MS MS-ESI (M/z)311.1[ M + H]⁺。

Intermediate: preparation of 6- (4-cyanopyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-11e

A yellow solid I-11e was prepared from intermediate I-11d (199.0mg,0.64mmol,1.0eq) and 80% hydrazine hydrate (60.0mg,0.96mmol,1.5eq) by a similar procedure as the third step in example 1. (by 100%). LC-MS MS-ESI (M/z)275.1[ M + H]⁺。

A compound: (E) preparation of (E) -6- (4-cyanopyridin-2-yl) -N' - (1- (pyridin-2-yl) ethylene) -2, 6-diazaspiro [3.3] hepta-2-thiohydrazide I-11

I-11 was prepared from I-11e (0.64mmol,1.0eq) and 2-acetylpyridine I-11f (85.2mg,0.70mmol,1.1eq) following a similar procedure as in compound I-3. (145.0mg, yield 60.1%). LC-MS MS-ESI (M/z)378.0[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.24(d,J＝5.0Hz,1H),7.93(d,J＝4.4Hz,1H),7.69(t,J＝7.1Hz,1H),7.53(d,J＝7.6Hz,1H),7.02–7.14(m,1H),6.76(d,J＝4.9Hz,1H),6.47(s,1H),4.36(dt,J＝28.8,14.3Hz,4H),4.21(s,4H),2.61(s,3H)。

Example 12

N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-12

Intermediate: preparation of 6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylic acid tert-butyl ester I-12c

Pale yellow solid I-12c was prepared from 2-bromopyridine I-1a (316.0mg,2.0mmol,1.0eq), 2, 6-diazaspiro [3.3]]Heptane-2-carboxylic acid tert-butyl hemioxalate I-10b (486.0mg,1.0mmol,0.5eq) was prepared by analogy with the first step in example 1. (280.0mg, yield 50.9%). LC-MS MS-ESI (M/z)276.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptan-2-yl) thione I-12d

A pale yellow solid I-12d was prepared from intermediate I-12c (280.0mg,1.02mmol,1.0eq), TFA (8mL), TEA (0.7mL) and TCDI (199.6mg,1.12mmol,1.1eq) following the second analogous procedure in example 1. (240.0mg, yield 82.5%). LC-MS MS-ESI (M/z)286.1[ M + H]⁺。

Intermediate: preparation of 6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-12e

A yellow solid I-12e was prepared from intermediate I-12d (240.0mg,0.84mmol,1.0eq) and 80% hydrazine hydrate (78.7mg,1.26mmol,1.5eq) following the third analogous procedure in example 1. (by 100%). LC-MS MS-ESI (M/z)250.1[ M + H]⁺。

A compound: preparation of N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-12

A yellow solid I-12 was prepared from I-12e (160.0mg,0.64mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (103.0mg,0.70mmol,1.1eq) by the fourth analogous procedure as in example 1. (80.0mg, yield 33.1%). LC-MS MS-ESI (M/z)379.2[ M + H [ ]]⁺. Compound I-12 is a mixture of formula E and formula Z, with a Z/E of about 1: 0.3.¹H-NMR(400MHz,CDCl₃)δppm 14.92(s,1H),8.57(d,J＝4.4Hz,1H),8.16(t,J＝5.8Hz,1H),7.60(d,J＝7.8Hz,1H),7.50-7.44(m,1H),7.24(d,J＝4.8Hz,1H),6.68-6.62(m,1H),6.32(d,J＝8.4Hz,1H),4.80(s,2H),4.48(s,2H),4.19(s,4H),2.89(t,J＝6.1Hz,2H),2.71-2.64(m,2H),1.99(d,J＝6.1Hz,2H)。

Example 13

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4- (trifluoromethyl) pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-13

Intermediate: preparation of 6- (4- (trifluoromethyl) pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylic acid tert-butyl ester I-13c

Pale yellow solid I-13c was prepared from 2-bromo-4- (trifluoromethyl) pyridine I-13a (450.0mg,2.0mmol,1.0eq), 2, 6-diazaspiro [3.3]]Heptane-2-carboxylic acid tert-butyl hemioxalate I-10b (486.0mg,1.0mmol,0.5eq) was prepared by analogy with the first step in example 1. (350.0mg, yield 51.0%). LC-MS MS-ESI (M/z)344.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (6- (4- (trifluoromethyl) pyridin-2-yl) -2, 6-diazaspiro [3.3] heptan-2-yl) methinone I-13d

A pale yellow solid I-13d was prepared from intermediate I-13c (350.0mg,1.02mmol,1.0eq), TFA (8mL), TEA (0.7mL) and TCDI (199.6mg,1.12mmol,1.1eq) following the second analogous procedure in example 1. (220.0mg, yield 61.1%). LC-MS MS MS-ESI (M/z)354.1[ M + H]⁺。

Intermediate: preparation of 6- (4- (trifluoromethyl) pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-13e

A pale yellow solid I-13e was prepared from intermediate I-13d (220.0mg,0.62mmol,1.0eq) and 80% hydrazine hydrate (78.7mg,1.26mmol,1.5eq) following the third analogous procedure in example 1. (140.0mg, yield 71.2%). LC-MS MS-ESI (M/z)318.1[ M + H]⁺. A compound: (Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4- (trifluoromethyl) pyridin-2-yl) -2, 6-diazaspiro [3.3]Preparation of heptane-2-thiohydrazide I-13

A yellow solid I-13 was prepared from I-13e (140.0mg,0.44mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (70.7mg,0.48mmol,1.1eq) by the fourth analogous procedure as in example 1. (60.0mg, yield 30.5%). LC-MS MS-ESI (M/z)447.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 14.94(s,1H),8.57(d,J＝4.2Hz,1H),8.38(s,1H),7.61(t,J＝6.5Hz,2H),7.24(d,J＝4.8Hz,1H),6.31(d,J＝8.7Hz,1H),4.80(s,2H),4.50(s,2H),4.26(s,4H),2.89(t,J＝6.1Hz,2H),2.74-2.64(m,2H),2.06-1.93(m,2H)。

Example 14

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4-methylpyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-14

Intermediate: preparation of 6- (4-methylpyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylic acid tert-butyl ester I-14c

Pale yellow solid I-14c was prepared from 2-bromo-4-methylpyridine I-14a (342.0mg,2.0mmol,1.0eq), 2, 6-diazaspiro [3.3]]Heptane-2-carboxylic acid tert-butyl hemioxalate I-10b (486.0mg,1.0mmol,0.5eq) was prepared by analogy with the first step in example 1. (230.0mg, yield 39.8%). LC-MS MS-ESI (M/z)290.2[ M + H [ ]]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (6- (4-methylpyridin-2-yl) -2, 6-diazaspiro [3.3] heptan-2-yl) methinone I-14d

A pale yellow solid I-14d was prepared from intermediate I-14c (230.0mg,0.79mmol,1.0eq), TFA (8mL), TEA (0.7mL) and TCDI (199.6mg,1.12mmol,1.1eq) following the second analogous procedure in example 1. (150.0mg, yield 63.5%). LC-MS MS MS-ESI (M/z)300.2[ M + H]⁺。

Intermediate: preparation of 6- (4-methylpyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-14e

Pale yellow solid I-14e was prepared from intermediate I-14d (150.0mg,0.50mmol,1.0eq) and 80% hydrazine hydrate (46.9mg,0.75mmol,1.5eq) following the third analogous procedure in example 1 to give the crude product. (100.0mg, yield 76.0%). LC-MS MS-ESI (M/z)264.2[ M + H]⁺. A compound: (Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4-methylpyridin-2-yl) -2, 6-diazaspiro [3.3]Preparation of heptane-2-thiohydrazide I-14

A yellow solid, I-14, was prepared from I-14e (100.0mg,0.38mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one, I-1f (61.8mg,0.42mmol,1.1eq) by the fourth analogous procedure as in example 1. (50.0mg, yield 33.5%). LC-MS MS-ESI (M/z)393.2[ M + H ]]⁺。¹H-NMR(400MHz,CDCl₃)δppm 14.92(s,1H),8.57(d,J＝3.7Hz,1H),8.02(d,J＝5.0Hz,1H),7.60(d,J＝7.3Hz,1H),7.23(s,1H),6.49(d,J＝4.8Hz,1H),6.14(s,1H),4.79(s,2H),4.47(s,2H),4.17(s,4H),2.88(t,J＝5.6Hz,2H),2.68(s,2H),2.26(s,3H),1.94-20.3(m,2H)。

Example 15

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4-methoxypyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-15

Intermediate: preparation of 6- (4-methoxypyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylic acid tert-butyl ester I-15c

Light yellow solid I-15c was prepared from 2-bromo-4-methoxypyridine I-15a (561.0mg,3.0mmol,1.0eq), 2, 6-diazaspiro [3.3]]Heptane-2-carboxylic acid tert-butyl hemioxalate I-10b (730.0mg,1.5mmol,0.5eq) was prepared by analogy with the first step in example 1. (420.0mg, yield 45.9%). LC-MS MS-ESI (M/z)306.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (6- (4-methoxypyridin-2-yl) -2, 6-diazaspiro [3.3] heptan-2-yl) methinone I-15d

A pale yellow solid I-15d was prepared from intermediate I-15c (420.0mg,1.38mmol,1.0eq), TFA (8mL), TEA (0.7mL) and TCDI (270.9mg,1.52mmol,1.1eq) following the second analogous procedure in example 1. (305.0mg, yield 70.2%). LC-MS MS-ESI (M/z)316.2[ M + H]⁺。

Intermediate: preparation of 6- (4-methoxypyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-15e

A pale yellow solid I-15e was prepared from intermediate I-15d (305.0mg,0.97mmol,1.0eq) and 80% hydrazine hydrate (90.9mg,1.46mmol,1.5eq) by following the third similar procedure in example 1. (210.0mg, yield 77.3%). LC-MS MS-ESI (M/z)280.2[ M + H [ ]]⁺。

A compound: preparation of (Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4-methoxypyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-15

Yellow solid I-15 is a solid prepared from I-15e (210.0mg,0.75mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (122.2mg,0.83mmol,1.1eq) were prepared by following the fourth analogous procedure in example 1. (150.0mg, yield 49.0%). LC-MS MS-ESI (M/z)409.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 14.90(s,1H),8.57(d,J＝3.4Hz,1H),7.99(d,J＝5.9Hz,1H),7.60(d,J＝7.9Hz,1H),7.25-7.22(m,1H),6.26(dd,J＝5.9,2.1Hz,1H),5.75(d,J＝2.0Hz,1H),4.78(s,2H),4.47(s,2H),4.17(s,4H),3.81(s,3H),2.88(t,J＝6.1Hz,2H),2.72-2.65(m,2H),2.01(dd,J＝12.4,6.3Hz,2H)。

Example 16

(Z) -6- (5-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-16

Intermediate: preparation of 6- (5-cyanopyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-carboxylic acid tert-butyl ester I-16c

Pale yellow solid I-16c was prepared from 2-bromo-5-cyanopyridine I-16a (316.0mg,2.0mmol,1.0eq), 2, 6-diazaspiro [3.3]]Heptane-2-carboxylic acid tert-butyl hemioxalate I-10b (486.0mg,1.0mmol,0.5eq) was prepared by analogy with the first step in example 1. (180.0mg, yield 30.0%). LC-MS MS-ESI (M/z)301.2[ M + H [ ]]⁺。

Intermediate: preparation of 6- (6- (1H-imidazole-1-thiocarbonyl) -2, 6-diazaspiro [3.3] heptan-2-yl) nicotinonitrile I-16d

A pale yellow solid I-16d was prepared from intermediate I-16c (180.0mg,0.58mmol,1.0eq), TFA (8mL), TEA (0.8mL) and TCDI (114.0mg,0.64mmol,1.1eq) following the second analogous procedure in example 1. (160.0mg, yield 88.9%). LC-MS MS-ESI (M/z)311.2[ M + H]⁺。

Intermediate: preparation of 6- (5-cyanopyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-16e

Pale yellow solid I-16e was prepared from intermediate I-16d (160.0mg,0.51mmol,1.0eq) and 80% hydrazine hydrate (48.1mg,0.77mmol,1.5eq) as in the third example 1The crude product is prepared by steps similar to the steps. (120.0mg, yield 85.8%). LC-MS MS-ESI (M/z)275.2[ M + H]⁺。

A compound: preparation of (Z) -6- (5-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-16

A yellow solid I-16 was prepared from I-16e (120.0mg,0.43mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (69.2mg,0.47mmol,1.1eq) by the fourth analogous procedure as in example 1. (75.0mg, yield 43.2%). LC-MS MS-ESI (M/z)404.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 14.95(s,1H),8.58(d,J＝4.6Hz,1H),8.40(d,J＝2.1Hz,1H),7.61(dd,J＝8.9,2.0Hz,2H),7.24-7.28(m,1H),6.27(d,J＝8.8Hz,1H),4.82(s,2H),4.51(s,2H),4.27(d,J＝16.9Hz,4H),2.89(t,J＝6.0Hz,2H),2.72-2.63(m,2H),2.01(dd,J＝12.2,6.1Hz,2H)。

Example 17

N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octane-7-thiohydrazide I-17

Intermediate: preparation of 4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octane-7-carboxylic acid tert-butyl ester I-17c

Pale yellow solid I-17c was prepared from 2-bromopyridine I-1a (1.8g,11.3mmol,1.0eq), 4, 7-diazaspiro [2.5]]Octane-7-carboxylic acid tert-butyl ester I-17b (1.2g,5.65mmol,0.5eq) was prepared by analogy with the first step in example 1. The crude product was separated by column chromatography on silica gel (PE/EA (v/v) ═ 4/1 elution), and then concentrated. (650.0mg, yield 20.0%). LC-MS MS-ESI (M/z)290.2[ M + H [ ]]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octan-7-yl) methylsulfan I-17d

A pale yellow solid I-17d was prepared from intermediate I-17c (121.1mg,0.42mmol,1.0eq), TFA (8mL), TEA (0.8mL) and TCDI (82.0mg,0.46mmol,1.1eq) following the second analogous procedure in example 1.(110.0mg, yield 87.8%). LC-MS MS-ESI (M/z)299.2[ M + H]⁺。

Intermediate: preparation of 4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octane-7-thiohydrazide I-17e

Pale yellow solid I-17e was prepared from intermediate I-17d (110.0mg,0.37mmol,1.0eq) and 80% hydrazine hydrate (35.0mg,0.56mmol,1.5eq) following the third analogous procedure in example 1. (70.0mg, yield 72.2%). LC-MS MS-ESI (M/z)263.2[ M + H]⁺。

A compound: preparation of N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octane-7-thiohydrazide I-17

A yellow solid I-17 was prepared from I-17e (70.0mg,0.26mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (42.7mg,0.29mmol,1.1eq) by the fourth analogous procedure as in example 1. (25.0mg, yield 23.9%). LC-MS MS-ESI (M/z)404.2[ M + H]⁺. Compound I-17 is a mixture of formula E and formula Z, with a Z/E of about 1: 0.14.¹H-NMR(400MHz,CDCl₃)δppm 14.76(s,1H),8.59(d,J＝4.0Hz,1H),8.23(d,J＝4.5Hz,1H),7.50(t,J＝7.6Hz,2H),7.22(dd,J＝7.7,4.6Hz,1H),7.07(d,J＝8.3Hz,1H),6.71-6.63(m,1H),4.15(s,6H),3.07(s,2H),2.88(t,J＝6.0Hz,2H),2.09-1.99(m,2H),1.14(d,J＝21.3Hz,2H),0.93(s,2H)。

Example 18

(Z) -6- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-18

A compound: preparation of (Z) -6- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-18

Yellow solid I-18 was prepared from I-11e (1.7g,6.20mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (1.0g,6.82mmol,1.1eq) by the fourth analogous procedure as in example 1. (1.6g, yield 64.0%). LC-MS MS-ESI (M/z)404.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 14.94(s,1H),8.57(d,J＝3.9Hz,1H),8.25(d,J＝5.0Hz,1H),7.60(d,J＝7.6Hz,1H),7.24(d,J＝4.9Hz,1H),6.78(d,J＝4.9Hz,1H),6.47(s,1H),4.75(d,J＝36.6Hz,4H),4.23(s,4H),2.88(t,J＝5.9Hz,2H),2.72-2.61(m,2H),2.06-1.92(m,2H)。

Example 19

(E) -6- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-19

A compound: (E) preparation of (E) -6- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-19

Yellow solid I-19 was prepared from I-11e (1.37g,5.0mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (809.6mg,5.5mmol,1.1eq) by the fourth analogous procedure as in example 1 to give compound I-18. Further subject I-18 to preparative TLC (DCM/CH)₃OH (v/v) ═ 15/1), and then concentrated. (1.1g, yield 54.5%). LC-MS MS-ESI (M/z)404.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.24(d,J＝5.1Hz,1H),7.78(d,J＝3.8Hz,1H),7.40(d,J＝7.3Hz,1H),6.98(dd,J＝7.7,4.9Hz,1H),6.75(d,J＝5.1Hz,1H),6.46(s,1H),4.41(d,J＝9.9Hz,2H),4.31(d,J＝10.2Hz,2H),4.25-4.14(m,4H),3.24-3.05(m,2H),2.83(t,J＝5.8Hz,2H),2.00(s,2H)。

Example 20

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-7-thiohydrazide I-20

Intermediate: preparation of 2- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester I-20c

Black solid I-20c was prepared from 2-bromopyridine I-1a (474.0mg,3.0 mm)ol,1.0eq), 2, 7-diazaspiro [3.5]]Nonane-7-carboxylic acid tert-butyl ester I-20b (678.0mg,3.0mmol,1.0eq) was prepared by analogy with the first step in example 1. (398.0mg, yield 43.8%). LC-MS MS MS-ESI (M/z)304.2[ M + H]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (2- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonan-7-yl) thione I-20d

A yellow solid I-20d was prepared from intermediate I-20c (398.0mg,1.31mmol,1.0eq) and TCDI (256.6mg,1.44mmol,1.1eq) following the second analogous procedure in example 1. (304.0mg, yield 74.1%). LC-MS MS-ESI (M/z)314.1[ M + H]⁺。

Intermediate: preparation of 4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octane-7-thiohydrazide 2- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-7-thiohydrazide I-20e

A yellow solid I-20e was prepared from intermediate I-20d (304.0mg,0.97mmol,1.0eq) and 80% hydrazine hydrate (91.0mg,1.46mmol,1.5eq) following the third analogous procedure in example 1. (by 100%). LC-MS MS-ESI (M/z)278.1[ M + H]⁺。

A compound: (E) preparation of (E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-7-thiohydrazide I-20

A yellow solid I-20 was prepared from I-20e (0.97mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (157.5mg,1.07mmol,1.1eq) following the fourth analogous procedure as in example 1. (153.0mg, yield 38.8%). LC-MS MS-ESI (M/z)407.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.14(d,J＝4.8Hz,1H),7.77(d,J＝4.4Hz,1H),7.43(t,J＝7.2Hz,1H),7.37(d,J＝7.2Hz,1H),7.00-6.92(m,1H),6.61-6.53(m,1H),6.29(d,J＝8.4Hz,1H),4.04(s,4H),3.79(s,4H),3.17(d,J＝6.0Hz,2H),2.82(s,2H),2.00(s,2H),1.86(d,J＝4.4Hz,4H)。

Example 21

(E) -6- (4-cyanopyridin-2-yl) -N' - ((3-fluoropyridin-2-yl) methylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-21

A compound: (E) preparation of (E) -6- (4-cyanopyridin-2-yl) -N' - ((3-fluoropyridin-2-yl) methylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-21

Yellow solid I-21 was prepared from I-11e (274.0mg,1.0mmol,1.0eq) and 3-fluoropyridinecarboxaldehyde I-21f (137.6mg,1.1mmol,1.1eq) following the fourth analogous procedure as in example 1. The mixture was further subjected to preparative TLC (DCM/CH)₃OH (v/v) ═ 15/1), and then concentrated. (55.0mg, yield 14.4%). LC-MS MS-ESI (M/z)382.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δ8.63(s,1H),8.25(d,J＝5.1Hz,1H),7.81(d,J＝4.7Hz,1H),7.39(t,J＝8.9Hz,1H),7.18-7.10(m,1H),6.78(d,J＝5.2Hz,1H),6.47(s,1H),4.44(s,2H),4.39(s,2H),4.22(s,4H)。

Example 22

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-ylamino) -2-azaspiro [3.3] heptane-2-thiohydrazide I-22

Intermediate: preparation of 6- (pyridin-2-ylamino) -2-azaspiro [3.3] heptane-2-carboxylic acid tert-butyl ester I-22c

Pale yellow solid I-22c was prepared from 2-bromopyridine I-1a (471.0mg,3.0mmol,1.0eq), 6-amino-2-azaspiro [3.3]]Heptane-2-carboxylic acid tert-butyl ester I-22b (636.0mg,3.0mmol,1.0eq) was prepared by analogy with the first step in example 1. (520.0mg, yield 60.0%). LC-MS MS-ESI (M/z)290.2[ M + H [ ]]⁺。

Intermediate: preparation of (1H-imidazol-1-yl) (6- (pyridin-2-ylamino) -2-azaspiro [3.3] heptan-2-yl) methinone I-22d

Pale yellow solid I-22d was prepared from intermediate I-22c (260.0mg,0.9mmol,1.0eq) and TCDI (176.4mg,0.99mmol,1.1eq) following the second analogous procedure in example 1. (145.0mg, yield 53.7%). LC-MS MS MS-ESI (M/z)300.2[ M + H]⁺。

Intermediate: preparation of 6- (pyridin-2-ylamino) -2-azaspiro [3.3] heptane-2-thiohydrazide I-22e

Pale yellow solid I-22e was prepared from intermediate I-22d (145.0mg,0.48mmol,1.0eq) and 80% hydrazine hydrate (45.0mg,0.72mmol,1.5eq) by the third similar procedure as in example 1. (85.0mg, yield 67.3%). LC-MS MS-ESI (M/z)264.2[ M + H]⁺。

A compound: preparation of (Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-ylamino) -2-azaspiro [3.3] heptane-2-thiohydrazide I-22

Yellow solid I-22 was prepared from I-22e (85.0mg,0.32mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (51.5mg,0.35mmol,1.1eq) by the fourth similar procedure as in example 1. (47.0mg, yield 37.4%). LC-MS MS-ESI (M/z)393.2[ M + H ]]⁺。¹H-NMR(400MHz,CDCl₃)δppm 14.85(s,1H),8.56(d,J＝4.0Hz,1H),8.08(d,J＝4.3Hz,1H),7.60(d,J＝7.6Hz,1H),7.43(s,1H),7.25-7.19(m,1H),6.64-6.54(m,1H),6.31(d,J＝8.3Hz,1H),4.65(s,2H),4.32(d,J＝46.2Hz,2H),4.12(dd,J＝14.2,7.0Hz,1H),2.89(t,J＝5.8Hz,2H),2.76(s,2H),2.70(s,2H),2.12(t,J＝9.6Hz,2H),2.06-1.93(m,2H)。

Example 23

(E) -2- (6, 7-dihydroquinolin-8 (5H) -ylidene) -N- (2- (pyridin-2-yl) -2-azaspiro [3.3] heptan-6-yl) hydrazine-1-thioamide I-23

Intermediate: preparation of (2- (pyridin-2-yl) -2-azaspiro [3.3] heptan-6-yl) carbamate I-23c

Pale yellow solid I-23c was prepared from 2-bromopyridine I-1a (471.0mg,3.0mmol,1.0eq), (2-azaspiro [3.3]]Heptane-6-yl) carbamate I-23b (636.0mg,3.0mmol,1.0eq) was prepared by analogy with the first step in example 1. (480.0mg, yield 55.3%). LC-MS MS-ESI (M/z)290.2[ M + H [ ]]⁺。

Intermediate: preparation of N- (2- (pyridin-2-yl) -2-azaspiro [3.3] heptan-6-yl) -1H-imidazole-1-thiohydrazide I-23d

Pale yellow solid I-23d was prepared from intermediate I-23c (260.0mg,0.9mmol,1.0eq) and TCDI (176.4mg,0.99mmol,1.1eq) following the second analogous procedure in example 1. (155.0mg, yield 57.5%). LC-MS MS MS-ESI (M/z)300.2[ M + H]⁺。

Intermediate: preparation of N- (2- (pyridin-2-yl) -2-azaspiro [3.3] heptan-6-yl) hydrazinothioamide I-23e

Pale yellow solid I-23e was prepared from intermediate I-23d (155.0mg,0.51mmol,1.0eq) and 80% hydrazine hydrate (48.1mg,0.77mmol,1.5eq) by the third similar procedure as in example 1. (80.0mg, yield 59.6%). LC-MS MS-ESI (M/z)264.2[ M + H]⁺。

A compound: (E) preparation of (E) -2- (6, 7-dihydroquinolin-8 (5H) -ylidene) -N- (2- (pyridin-2-yl) -2-azaspiro [3.3] heptan-6-yl) hydrazine-1-thioamide I-23

Yellow solid I-23 was prepared from I-23e (80.0mg,0.3mmol,1.0eq) and 6, 7-dihydroquinolin-8 (5H) -one I-1f (48.6mg,0.33mmol,1.1eq) by the fourth analogous procedure as in example 1. The mixture was further subjected to preparative TLC (DCM/CH)₃OH (v/v) ═ 15/1), and then concentrated. (39.0mg, yield 33.1%). LC-MS MS-ESI (M/z)393.2[ M + H ]]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.12(d,J＝4.5Hz,1H),7.78(d,J＝4.4Hz,1H),7.42(t,J＝8.4Hz,2H),7.00(dd,J＝7.4,5.1Hz,1H),6.60-6.50(m,1H),6.25(d,J＝8.3Hz,1H),5.63(s,1H),4.64(s,1H),4.06(s,2H),3.94(s,2H),3.15(s,2H),2.83(s,2H),2.67-2.75(m,2H),2.16(t,J＝8.8Hz,2H),2.08-1.93(m,2H)。

Example 24

N' - (bis (pyridin-2-yl) methylene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-24

A compound: preparation of N' - (di (pyridin-2-yl) methylene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-24

Intermediate I-12e (160.0mg,0.64mmol,1.0eq) was dissolved in EtOH (15mL), bis (pyridin-2-yl) methanone (130.0mg,0.70mmol,1.1eq) and glacial acetic acid (1 drop) were added and the resulting solution heated to 95 ℃ and stirred for 8h, cooled to ambient temperature. The solid was collected by filtration and washed 1 time with MeOH (20mL) followed by EA (20mL) in a slurry to give I-24 as a yellow solid. (120mg, yield 45.1%). LC-MS MS-ESI (M/z)416.2[ M + H [ ]]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.78(d,J＝4.6Hz,1H),8.13(d,J＝4.6Hz,2H),7.93(d,J＝7.9Hz,1H),7.85(t,J＝6.9Hz,1H),7.56(t,J＝7.8Hz,1H),7.43(dd,J＝11.2,4.4Hz,1H),7.37-7.30(m,2H),7.14-7.05(m,1H),6.63-6.57(m,1H),6.28(d,J＝8.3Hz,1H),4.43(s,1H),4.20(s,3H),4.12(s,4H)。

Example 25

6- (4-cyanopyridin-2-yl) -N' - (bis (pyridin-2-yl) methylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-25

A compound: preparation of 6- (4-cyanopyridin-2-yl) -N' - (bis (pyridin-2-yl) methylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide I-25

Yellow solid I-25 was prepared from intermediate I-11e (274.0mg,1.0mmol,1.0eq) and bis (pyridin-2-yl) methanone (202.6mg,1.1mmol,1.1eq) following a similar procedure for compound I-24. (160.0mg, yield 36.3%). LC-MS MS-ESI (M/z)441.2[ M + H]⁺。¹H-NMR(400MHz,CDCl₃)δppm 8.79(d,J＝4.8Hz,1H),8.23(d,J＝5.1Hz,1H),8.13(d,J＝5.0Hz,1H),7.91(d,J＝7.7Hz,1H),7.86(t,J＝7.5Hz,1H),7.58(t,J＝7.6Hz,1H),7.39-7.34(m,1H),7.31(d,J＝8.2Hz,1H),7.14-7.08(m,1H),6.76(d,J＝5.0Hz,1H),6.44(s,1H),4.45(s,1H),4.22(s,3H),4.16(s,4H)。

In vitro biological evaluation

The detection method is used for evaluating the biological activity of the compound of the invention at a cellular level, and comprises a method for evaluating the biological activity of the compound in four cell line models with different p53 states.

The purpose of this assay was to evaluate the effect of different compounds on the biological activity of wild-type or mutated at different sites p53 protein, i.e., whether a compound could reactivate its biological functions of inhibiting tumor cell cycle and promoting tumor cell apoptosis by altering the conformation of the mutated p53 protein.

EXAMPLE A detection of half the growth inhibitory dose of Compounds on tumor cell lines of different p53 status

Principle of experiment

P53 is a well-known oncostatin, whose major biological functions are cell cycle inhibition and apoptosis induction, and the mutation of P53 is relatively common in tumor cells, and the loss of the biological function of P53 is a driving factor of cell canceration and tumor resistance.

Amino acid positions in P53, including but not limited to R175, Y220, G245, R248, R249, R273 and R282, often have missense mutations that prevent the P53 protein from forming the correct conformation and thus losing its biological function.

Four tumor cell lines used in the detection method respectively express wild p53 protein and three different forms of missense mutant p53 protein. Wherein the conformation and biological function of the p53 protein in the wild tumor cell line are normal, but the tumor cell can tolerate the periodic inhibition and apoptosis induction of the p53 protein; the p53 protein in the mutant tumor cell line fails to exert its biological function due to the conformational abnormality. Theoretically, if the test compound has and only has the function of restoring the correct conformation of the mutant p53 protein, when it acts on tumor cells expressing wild-type p53 protein, it does not exhibit killing of tumor cells, and when it acts on tumor cells expressing missense mutant p53 protein as described above, it can reactivate the killing of tumor cells by restoring the conformation of p53 protein. Thus, by detecting and comparing half the growth inhibitory dose of a compound on tumor cell lines of different p53 status, it can be assessed whether the compound is a selective mutant p53 reactivation agent.

Experimental materials and apparatus

The human non-small cell lung cancer cell line A549, the human breast cancer cell line MDA-MB-468, the human small cell lung cancer cell line NCI-H1048 and NCI-H1417 were purchased from American Type Culture Collection (ATCC), and the relevant Culture conditions were all referred to from ATCC. Wherein, A549 expresses wild type (wild type, wt) p53 protein; MDA-MB-468, NCI-H1048 and NCI-H1417 express mutant type (mut) p53 proteins at the mutation sites R273H, R273C and R175L, respectively. Alamar Blue^TMCell viability assay reagents were purchased from Sigma-Aldrich. The Forma Steri-Cycle cell culture box was purchased from Thermo Fisher Scientific.

Main process of experiment

Cell lines A594, MDA-MB-468 and NCI-H1048 cultured by adherence are digested by pancreatin to prepare cell suspension, and cell lines NCI-H1417 cultured by suspension are repeatedly sucked and beaten by a pipette to prepare cell suspension. The cell suspension was sampled and counted, and inoculated into 96-well cell culture plates at 5000/190. mu.L/well in 5% CO₂After overnight incubation in a 37 ℃ incubator, the test compound-broth mixtures were added at different concentrations in volumes of 10. mu.L per well, to give final test compound concentrations of 0,0.003,0.01,0.03,0.1,0.3,1,3,10, 30. mu.M per well, with 3 replicates per cell line and per test drug dose setting. After further culturing for 72 hours, alamar Blue was added according to the manual of the relevant product^TMCell viability detection reagent for detecting cell viability of different cell lines cultured under the action of different doses of test compounds, calculating the percentage of cell growth inhibition of each well by taking 0 mu M dose wells as reference, and further calculating the half growth inhibition dose (GI) of each test compound to different test cell lines by GraphPad Prism 7.00 software₅₀)。

GI₅₀Lower values indicate a more sensitive test cell line to the test compound, and higher values indicate a more resistant test cell line to the test compound. Theoretically, the A549 cell line expressing wild-type p53 protein was insensitive to selective p53 mutant allosteric activators, while the MDA-MB-468, NCI-H1048, and NCI-H1417 cell lines expressing mutant p53 protein were sensitive to selective p53 mutant allosteric activatorsThe greater the difference in potency, the more selective the test compound.

Based on the above test methods, the activity data of the compounds according to the invention are summarized (see table 1).

TABLE 1 GI of the Compounds of the invention in p53 wild-type or mutant cell lines₅₀(μM)

The results show that the compound of the invention has good in vitro inhibitory activity on tumor cell lines with different p53 states, and the compound of the general formula (I) of the invention also has the inhibitory activity.

Claims

1. A compound of the formula (I),

or a pharmaceutically acceptable salt or isomer thereof,

wherein the content of the first and second substances,

R¹selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl (e.g. furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl) which is a 5-to 12-membered monocyclic or polycyclic, wherein the heteroaryl may be substituted with one or more substituents independently selected from deuterium, halogen (e.g. fluorine, chlorine, bromine or iodine), C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino, di-C₁-C₄Radical substitution of alkylamino;

R²selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl (e.g. furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl) which is a 5-to 12-membered monocyclic or polycyclic, wherein the heteroaryl may be substituted with one or more substituents independently selected from deuterium, halogen (e.g. fluorine, chlorine, bromine or iodine), C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino, di-C₁-C₄Radical substitution of alkylamino;

alternatively, the first and second electrodes may be,

R¹and R²Together with the carbon atom to which they are commonly attached form a heteroaryl (e.g., dihydroquinolinyl) or a 3-15 membered mono-or polycyclic heterocycle, including but not limited to spirocyclic, bridged, wherein said 3-15 membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from the group consisting of halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to an aryl or heteroaryl group;

R³selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, aryl, arylamino, heteroaryl, heteroarylamino, heteroarylaminoaryl (said heteroaryl being, for example, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl; said aryl being, for example, phenyl), 3-15 membered mono-or polycyclic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen or sulfur (for example, azaspiro [3.3]]Heptane-6-yl), 3-15 membered mono-or polycyclic heterocyclic amino, wherein said arylamino, heteroarylamino, 3-15 membered mono-or polycyclic heterocyclic amino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl; r⁴Selected from hydrogen, deuterium, C₁-C₄Alkylamino, halogeno C₁-C₄Alkylamino, arylamino, heteroarylamino, heteroarylaminoaryl (said heteroaryl being, for example, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl; said aryl being, for example, phenyl), 3-15 membered mono-or polycyclic heterocycle containing one or two heteroatoms selected from nitrogen, oxygen or sulfur (for example, azaspiro [3.3]]Heptane-6-yl), 3-15 membered mono-or polycyclic heterocyclic amino, wherein said C₁-C₄Alkylamino, halogeno C₁-C₄Alkylamino, arylamino, heteroarylamino, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

R³and R⁴Together with the nitrogen atom to which they are both attached form a 3-15 membered mono-or polycyclic heterocycle optionally containing one additional heteroatom selected from nitrogen, oxygen or sulfur, wherein said 3-15 membered mono-or polycyclic heterocycle may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

R^aselected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, aryl, heteroaryl (e.g., furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl; benzofuryl, benzimidazolyl, benzisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothienyl, benzotriazolyl, benzoxazolyl, furopyridyl, imidazopyridinyl, imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuryl, isobenzothienyl, isoindolyl, isoquinolyl, dihydroquinolinyl, naphthyridinyl, oxazolopyridyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quinazolinyl, pyrazinyl, pyrazolyl, pyridazinyl, benzothiazolyl, benzimidazolyl, benzoxazolyl, benzothienyl, isoindolyl, isoquinolinyl, dihydroquinolinyl, naphthyridinyl, oxazolopyridyl, phthalazinyl, pteridinyl, purinyl, pyridopyridyl, pyrrolopyridyl, quinolyl, and the like, Thiadiazolopyrimidinyl and thienopyridinyl; preferably pyridyl, quinolyl), heteroarylamino (said heteroaryl being for example furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl or triazolyl, preferably pyridyl), said heteroaryl being a 5-12 membered monocyclic or polycyclic, wherein said aryl, heteroaryl may be substituted by one or more groups independently selected from hydrogen, deuterium, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkoxy, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino radical, mono C₁-C₄Alkylamino radical C₁-C₄Alkyl, di-C₁-C₄Alkylamino radical C₁-C₄Alkyl groups.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

R¹selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring, wherein said heteroaryl may be substituted by one or more independently selected from deuterium, halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino, di-C₁-C₄Radical substitution of alkylamino;

R²selected from hydrogen, deuterium, C₁-C₄Alkyl, halo C₁-C₄Alkyl, heteroaryl, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring, wherein said heteroaryl may be substituted by one or more independently selected from deuterium, halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxy, CONH₂Sulfonamide, aminosulfonyl, mono C₁-C₄Alkylamino, di-C₁-C₄Radical substitution of alkylamino;

alternatively, the first and second electrodes may be,

R¹and R²Together with the carbon atom to which they are commonly attached form a heteroaryl (e.g., dihydroquinolinyl) or a 3-15 membered mono-or polycyclic heterocyclic ring, including but not limited to spirocyclic heterocyclic ringsBridged heterocycles, wherein the 3-15 membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to an aryl or heteroaryl group;

R³selected from hydrogen, deuterium;

R⁴selected from arylamino, heteroarylamino, heteroarylaminoaryl, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino, wherein said arylamino, heteroarylamino, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

R³and R⁴Together with the nitrogen atom to which they are both attached form a 3-15 membered mono-or polycyclic heterocycle, wherein the 3-15 membered mono-or polycyclic heterocycle may be substituted with one or more R^aSubstituted, said 3-15 membered polycyclic heterocycle includes but is not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

R²selected from heteroaryl, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring, wherein said heteroaryl may be substituted by one or more substituents independently selected from halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, hydroxy, mercapto, nitro, carboxyl, mono C₁-C₄Alkylamino, di-C₁-C₄Radical substitution of alkylamino;

alternatively, the first and second electrodes may be,

R³selected from hydrogen, deuterium;

R⁴selected from the group consisting of heteroarylamino, heteroarylaminoaryl, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino, wherein said heteroarylamino, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino may be substituted with one or more R^aSubstituted, said 3-15 membered polycyclic heterocyclic ring including but not limited toLimited to a spirocyclic heterocycle, a bridged heterocycle, wherein the spirocyclic heterocycle or the bridged heterocycle may be fused to an aryl or heteroaryl group;

alternatively, the first and second electrodes may be,

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

R¹selected from hydrogen, deuterium, C₁-C₄Alkyl, heteroaryl, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring, wherein said heteroaryl may be substituted by one or more independently selected from deuterium, halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, mono C₁-C₄Alkylamino, di-C₁-C₄Radical substitution of alkylamino;

R²selected from heteroaryl, said heteroaryl being a 5-12 membered monocyclic or polycyclic ring, wherein said heteroaryl may be substituted by one or more substituents independently selected from halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, mono C₁-C₄Alkylamino, di-C₁-C₄Radical substitution of alkylamino; alternatively, the first and second electrodes may be,

R¹and R²Together with the carbon atom to which they are both attached form a heteroaryl group (e.g., dihydroquinolinyl) or a 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from the group consisting of halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to aryl or heteroaryl, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle;

R³selected from hydrogen, deuterium;

R⁴selected from the group consisting of heteroarylaminoaryl, 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino, wherein said 3-15 membered mono-or polycyclic heterocycle, 3-15 membered mono-or polycyclic heterocyclylamino may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

R¹selected from hydrogen, deuterium, C₁-C₄An alkyl group;

R²selected from pyridyl, wherein said pyridyl may be independently selected from one or more of halogen, C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkyl, halo C₁-C₄Alkoxy, cyano, amino, mono C₁-C₄Alkylamino, di-C₁-C₄Alkyl ammoniaRadical substitution of radicals;

alternatively, the first and second electrodes may be,

R³selected from hydrogen, deuterium;

R⁴selected from the group consisting of heteroarylaminoaryl, 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl;

alternatively, the first and second electrodes may be,

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

R¹and R²Together with the carbon atom to which they are commonly attached form a heteroaryl (e.g. dihydroquinolinyl) or 3-15 membered mono-or polycyclic heterocycle wherein said 3-1The 5-membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to aryl or heteroaryl, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle;

R³selected from hydrogen, deuterium;

R⁴selected from the group consisting of heteroarylaminoaryl, 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle may be substituted with one or more R^a(ii) substituted, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle, wherein spirocyclic heterocycle or bridged heterocycle may be fused to aryl or heteroaryl; alternatively, the first and second electrodes may be,

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

R¹and R²Together with the carbon atom to which they are both attached form a heteroaryl group (e.g., dihydroquinolinyl) or a 3-15 membered mono-or polycyclic heterocycle, wherein said 3-15 membered mono-or polycyclic heterocycle is optionally substituted with one or more substituents selected from the group consisting of halogen, amino, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl, hydroxy C₁-C₄Alkyl, amino C₁-C₄Alkyl radical, C₁-C₄Alkoxy, monoC₁-C₄Alkylamino, di-C₁-C₄Alkylamino, wherein said 3-15 membered mono-or polycyclic heterocycle may be fused to aryl or heteroaryl, said 3-15 membered polycyclic heterocycle including but not limited to spirocyclic heterocycle, bridged heterocycle;

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof,

wherein the content of the first and second substances,

R^aselected from aryl, heteroaryl, heteroArylamino, wherein said aryl, heteroaryl may be substituted with one or more substituents independently selected from hydrogen, deuterium, cyano, C₁-C₄Alkyl, halo C₁-C₄Alkyl radical, C₁-C₄Alkoxy, halo C₁-C₄Alkoxy, mono C₁-C₄Alkylamino, di-C₁-C₄Alkylamino radical, mono C₁-C₄Alkylamino radical C₁-C₄Alkyl, di-C₁-C₄Alkylamino radical C₁-C₄Alkyl, and the heteroaryl is a 5-12 membered monocyclic or polycyclic ring.

9. The compound according to any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein R³And R⁴A 3-15 membered mono-or polycyclic heterocyclic ring formed together with the nitrogen atom to which they are both attached, said 3-15 membered mono-or polycyclic heterocyclic ring selected from:

10. a compound according to any one of claims 1 to 9, selected from a single configuration or a mixture of isomers thereof, selected from:

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -9- (pyridin-2-yl) -3, 9-diazaspiro [5.5] undecane-3-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -9- (pyridin-2-yl) -3, 9-diazaspiro [5.5] undecane-3-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -1, 4-diazepan-1-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -1, 4-diazepan-1-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -7- (pyridin-2-yl) -2, 7-diazaspiro [4.4] nonane-2-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -7- (pyridin-2-yl) -2, 7-diazaspiro [4.4] nonane-2-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -7- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -7- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide;

(3aR,6aS) -N' - ((E) -6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -thiohydrazide;

(3aR,6aS) -N' - ((Z) -6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -thiohydrazide;

(E) -2- (6, 7-dihydroquinolin-8 (5H) -ylidene) -N- (4- (pyridin-2-ylamino) phenyl) hydrazine-1-thioamide;

(Z) -2- (6, 7-dihydroquinolin-8 (5H) -ylidene) -N- (4- (pyridin-2-ylamino) phenyl) hydrazine-1-thioamide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) -1, 5-diazacyclooctane-1-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) -1, 5-diazacyclooctane-1-thiohydrazide;

(E) -7- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide;

(Z) -7- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 7-diazaspiro [3.5] nonane-2-thiohydrazide;

(E) -4- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -1, 4-diazepan-1-thiohydrazide;

(Z) -4- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -1, 4-diazepan-1-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (quinolin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (quinolin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -6- (4-cyanopyridin-2-yl) -N' - (1- (pyridin-2-yl) ethylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -6- (4-cyanopyridin-2-yl) -N' - (1- (pyridin-2-yl) ethylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4- (trifluoromethyl) pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4- (trifluoromethyl) pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4-methylpyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4-methylpyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4-methoxypyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (4-methoxypyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -6- (5-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -6- (5-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octane-7-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octane-7-thiohydrazide;

(E) -6- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -6- (4-cyanopyridin-2-yl) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-7-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -2- (pyridin-2-yl) -2, 7-diazaspiro [3.5] nonane-7-thiohydrazide;

(E) -6- (4-cyanopyridin-2-yl) -N' - ((3-fluoropyridin-2-yl) methylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -6- (4-cyanopyridin-2-yl) -N' - ((3-fluoropyridin-2-yl) methylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(Z) -N' - ((3-aminopyridin-2-yl) methylene) -6- (4-cyanopyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

(E) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-ylamino) -2-azaspiro [3.3] heptane-2-thiohydrazide;

(Z) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-ylamino) -2-azaspiro [3.3] heptane-2-thiohydrazide;

(E) -2- (6, 7-dihydroquinolin-8 (5H) -ylidene) -N- (2- (pyridin-2-yl) -2-azaspiro [3.3] heptan-6-yl) hydrazine-1-thioamide;

(Z) -2- (6, 7-dihydroquinolin-8 (5H) -ylidene) -N- (2- (pyridin-2-yl) -2-azaspiro [3.3] heptan-6-yl) hydrazine-1-thioamide;

n' - (bis (pyridin-2-yl) methylene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

6- (4-cyanopyridin-2-yl) -N' - (bis (pyridin-2-yl) methylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide.

11. A compound according to any one of claims 1 to 9, selected from the group consisting of

Compound I-1: n' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -9- (pyridin-2-yl) -3, 9-diazaspiro [5.5] undecane-3-thiohydrazide;

compound I-5: (3aR,6aS) -N' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) hexahydropyrrolo [3,4-c ] pyrrole-2 (1H) -thiohydrazide;

compound I-7: n' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -5- (pyridin-2-yl) -1, 5-diazacyclooctane-1-thiohydrazide;

compound I-12: n' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

compound I-17: n' - (6, 7-dihydroquinolin-8 (5H) -ylidene) -4- (pyridin-2-yl) -4, 7-diazaspiro [2.5] octane-7-thiohydrazide;

compound I-25: n' - (bis (pyridin-2-yl) methylene) -6- (pyridin-2-yl) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide;

compound I-26: 6- (4-cyanopyridin-2-yl) -N' - (bis (pyridin-2-yl) methylene) -2, 6-diazaspiro [3.3] heptane-2-thiohydrazide.

12. A process for preparing a compound according to any one of claims 1 to 11, characterized in that:

carrying out one-pot reaction on the compound shown in the formula (I-c) to obtain a compound shown in the formula (I-d), wherein the deprotection is carried out by using an acid or alkali reagent under the condition of a proper solvent in the 1 st stage; the 2 nd stage is to react with N, N' -thiocarbonyl diimidazole (TCDI) under the catalysis of a proper alkali reagent under the condition of a proper solvent to obtain a compound shown as a formula (I-d);

reacting the compound shown in the formula (I-d) with hydrazine hydrate under the condition of a proper solvent to obtain a compound shown in the formula (I-e);

reacting the compound shown in the formula (I-e) with the compound shown in the formula (I-f) under the condition of a proper solvent to obtain the compound shown in the formula (I);

13. The method according to claim 12, wherein the compound of formula (I-c) is obtained by Buchwald-Hartwig coupling reaction under appropriate alkaline conditions and in a solvent and in the presence of a catalyst,

the catalyst is a palladium reagent including, but not limited to, palladium acetate (Pd (OAc)₂) 1,1 '-binaphthyl-2, 2' -bis (diphenylphosphine) (BINAP), 2-dicyclohexylphosphine-2 ',6' -dimethoxybiphenyl (SPhos), dicyclohexyl (3, 6-dimethoxy-2 ',4',6 '-triisopropyl (1,1' -biphenyl) -2-yl) phosphine (Brettphos), tris (dibenzylideneacetone) dipalladium (Pd)₂(dba)₃) [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (PdCl)₂(dppf)), [1,1' -bis (diphenylphosphino) ferrocene]Palladium dichloride dichloromethane complex (PdCl)₂(dppf)·CH₂Cl₂) Tetrakis (triphenylphosphine) palladium (Pd (PPh)₃)₄) Bis (tricyclohexylphosphine) palladium dichloride (PdCl)₂(P(Cy)₃)₂)；

Such solvents include, but are not limited to, Toluene (Toluene), 1,4-dioxane (1,4-dioxane), Tetrahydrofuran (THF), acetonitrile (CH)₃CN), N' -Dimethylformamide (DMF), and mixed solvents of these solvents in different ratios to water.

14. The method according to claim 12, wherein the compound of formula (I-c) is obtained by Buchwald-Hartwig coupling reaction under proper alkaline conditions and solvent and under catalyst, or nucleophilic substitution reaction under proper alkaline conditions and solvent,

the basic condition reagent includes organic and inorganic bases including, but not limited to, Triethylamine (TEA), N-Diisopropylethylamine (DIPEA), N-butyllithium, lithium diisopropylamide, lithium bistrimethylsilylamide, potassium acetate (KOAc), sodium t-butoxide (tBuONa), potassium t-butoxide (tBuOK), sodium hydride (NaH), potassium phosphate (K)₃PO₄) Carbonic acidSodium (Na)₂CO₃) Potassium carbonate (K)₂CO₃) Lithium hydroxide (KOH), sodium hydroxide (NaOH).

15. The method according to claim 12, wherein the compound of formula (I-c) is obtained by nucleophilic substitution reaction under appropriate basic conditions and solvent,

such solvents include, but are not limited to, N' -Dimethylformamide (DMF), N-methylpyrrolidinone (NMP), 1,4-dioxane (1,4-dioxane), Tetrahydrofuran (THF), acetonitrile (CH)₃CN)。

16. The process of claim 12, wherein the compound of formula (I-c) is subjected to a one-pot reaction to obtain the compound of formula (I-d), comprising deprotection in stage 1 with an acid or base reagent under suitable solvent conditions; the 2 nd stage is to react with N, N' -Thiocarbonyldiimidazole (TCDI) under the catalysis of a proper alkali reagent under the condition of a proper solvent to obtain the compound shown in the formula (I-d),

such solvents include, but are not limited to, Dichloromethane (DCM), 1,4-dioxane (1,4-dioxane) Tetrahydrofuran (THF), acetonitrile (CH)₃CN), N' -Dimethylformamide (DMF);

the acid reagents include, but are not limited to, trifluoroacetic acid (TFA) and hydrochloric acid (HCl); the alkali agent includes, but is not limited to piperidine and diethylamine.

17. The method according to claim 12, wherein the compound of formula (I-d) is reacted with hydrazine hydrate under suitable solvent conditions to give the compound of formula (I-e), and the compound of formula (I-e) is reacted with the compound of formula (I-f) under suitable solvent conditions to give the compound of formula (I),

the solvent includes but is not limited to methanol (CH)₃OH), ethanol (EtOH).

18. A pharmaceutical composition comprising a compound of formula (I) as claimed in any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant.

19. Use of a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment and/or prevention of a disease associated with abnormal activity of p53 protein.

20. The use of claim 19, wherein the disease associated with abnormal activity of p53 protein comprises tumor or cancer.